CN117794560A - Stapled peptides and methods thereof - Google Patents

Stapled peptides and methods thereof Download PDF

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Publication number
CN117794560A
CN117794560A CN202280053893.9A CN202280053893A CN117794560A CN 117794560 A CN117794560 A CN 117794560A CN 202280053893 A CN202280053893 A CN 202280053893A CN 117794560 A CN117794560 A CN 117794560A
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agent
staple
independently
amino acid
group
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布赖恩·哈尔伯特·怀特
丹尼尔·圣杜克·拉
洛伦佐·若苏埃·阿尔法罗-洛佩斯
保拉·克里斯蒂娜·奥尔泰特
莎拉·伊莎贝尔·卡普奇
李志�
约翰·汉尼·麦吉
马丁·罗伯特·特朗布莱
格雷戈里·L·韦尔迪
司亚光
凯文·凌
杜培成
乔纳森·巴里·胡罗夫
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Vogel Pharmaceutical Co ltd
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Vogel Pharmaceutical Co ltd
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Priority claimed from PCT/US2022/032738 external-priority patent/WO2022261257A1/en
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Abstract

The present disclosure provides, inter alia, a variety of useful medicaments. In some embodiments, the provided agents can bind to β -catenin. In some embodiments, the present disclosure provides techniques for modulating β -catenin function. In some embodiments, the present disclosure provides techniques for preventing and/or treating a condition, disorder or disease associated with β -catenin. In some embodiments, the present disclosure provides engineered amino acids and agents that can provide improved properties and/or activity.

Description

Stapled peptides and methods thereof
Cross Reference to Related Applications
The present application claims priority from U.S. provisional application No.63/208,487, filed on 6.08, 2021, U.S. provisional application No.63/224,834, filed on 7.22, 2021, and U.S. provisional application No.63/303,952, filed on 27, 2022, each of which is incorporated herein by reference in its entirety.
Background
Stapled peptides (stapled peptides) are useful in a variety of applications. For example, as bioactive agents, they can be used to modulate a variety of biological functions.
Disclosure of Invention
The present disclosure provides, inter alia, powerful techniques (e.g., agents (e.g., peptides or those comprising peptides, which in many embodiments are stapled peptides), compositions, methods, etc.) for modulating a variety of biological functions.
In some embodiments, the present disclosure provides pharmaceutical agents, such as stapled peptides comprising a plurality of staple-like structures. In some embodiments, the present disclosure provides pharmaceutical agents, e.g., stapled peptides comprising three or more staple-like structures. In some embodiments, the present disclosure provides agents, e.g., stapled peptides comprising three or more staple-like structures within 10 to 20 amino acid residues, e.g., 10 to 15, 11 to 14, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 consecutive amino acid residues. In some embodiments, the present disclosure provides agents, e.g., stapled peptides comprising three or more staple-like structures within 11 consecutive amino acid residues. In some embodiments, the present disclosure provides agents, e.g., stapled peptides comprising three or more staple-like structures within 14 consecutive amino acid residues. In some embodiments, there are three staple-like structures within such a number of amino acid residues. In some embodiments, there are four staple-like structures within such a number of consecutive amino acid residues. Without intending to be limited by theory, in some embodiments, the provided agents (e.g., stapled peptides) have increased rigidity as compared to a reference peptide (e.g., non-stapled peptides, or stapled peptides having fewer staple-like structures (in some embodiments, fewer staple-like structures within a certain number of amino acid residues, as described herein), etc. In some embodiments, the provided agents (e.g., stapled peptides) exhibit a variety of desirable properties and/or activities. In some embodiments, the provided agents (e.g., stapled peptides) provide improved desirable properties and/or activity compared to a reference peptide (e.g., non-stapled peptide, or a stapled peptide with fewer staple-like structures (in some embodiments, fewer staple-like structures within a certain number of amino acid residues, as described herein), etc.
In some embodiments, the provided technology comprises engineered structural features, e.g., new amino acid residues, that can provide significantly improved properties and/or activity compared to comparable reference technologies that do not comprise such engineered structural features. In some embodiments, the present disclosure provides engineered amino acids as described herein, which are incorporated into peptide agents (including stapled peptides), which can provide significantly improved properties and/or activity (e.g., improved lipophilicity) and/or delivery into cells compared to a reference amino acid (e.g., asp). In some embodiments, the present disclosure provides techniques including peptides comprising such engineered amino acid residues. In some embodiments, the present disclosure provides stapled peptides comprising such designed amino acid residues.
In some embodiments, the present disclosure provides techniques for modulating one or more functions of β -catenin. In particular, in some embodiments, the present disclosure provides a variety of agents, such as peptides, in many cases stapled peptides, that can bind to and modulate the function of β -catenin. As shown herein, in some embodiments, the binders (binders agents) of the present disclosure may interact with β -catenin at unique sets of residues. In some embodiments, the binding site comprises one or more or all of the groups of residues. In some embodiments, the provided agents interact with one or more of the following or groups of residues corresponding to: SEQ ID NO:1, the following residues: a305 Y306, G307, N308, Q309, K312, R342, K345, V346, V349, Q375, R376, Q379, N380, L382, W383, R386, N387, D413, N415, V416, T418, and C419. In some embodiments, the provided agents interact with one or more of the amino acid residues that are or correspond to: SEQ ID NO:1, a305, Y306, G307, N308, Q309, K312, R342, K345, V346, V349, Q375, Q379, N380, L382, W383, R386, N387, D413, N415, V416, T418, and C419. In some embodiments, the provided agents interact with one or more of the amino acid residues that are or correspond to: SEO ID NO:1, a305, Y306, G307, N308, Q309, K312, K345, V346, V349, Q379, N380, L382, W383, R386, N387, D413, N415, V416, T418, and C419. In some embodiments, the provided agents interact with one or more of the amino acid residues that are or correspond to: SEQ ID NO: g307, K312, K345, W383, N387, D413, and N415 of 1. In some embodiments, the provided agents interact with one or more of the amino acid residues that are or correspond to: SEQ ID NO: k312, K345, R386 and W383 of 1. In some embodiments, the provided agents interact with one or more of the following or groups of residues corresponding to: SEQ ID NO:1, the following residues: g307 K312, K345, Q379, L382, W383, N387, N415, and V416. In some embodiments, the provided agent interacts with all of the following or groups of residues corresponding to: SEQ ID NO:1, the following residues: y306, G307, K312, K345, Q379, L382, W383, N387, N415, and V416. In some embodiments, the provided agent interacts with all of the following or groups of residues corresponding to: SEQ ID NO:1, the following residues: g307 K312, K345, Q379, L382, W383, N387, N415, and V416. In some embodiments, the provided agent interacts with all of the following or groups of residues corresponding to: SEQ ID NO:1, the following residues: y306, G307, K312, K345, Q379, L382, W383, N387, N415, and V416. In some embodiments, the provided agents interact with one or more of the amino acid residues that are or correspond to: SEQ ID NO: k312, K345 and W383 of 1. In some embodiments, the provided agents interact with amino acid residues that are or correspond to: SEQ ID NO: k312, K345 and W383 of 1.
As described herein, the provided techniques can modulate one or more biological processes associated with β -catenin. In some embodiments, the provided agent (e.g., a stapled peptide) competes with a ligand (e.g., with a member of the T cell factor/lymphokine (TCF/LEF) family of transcription factors) for binding to β -catenin. In some embodiments, the provided agents compete with the ligand for binding to β -catenin at a particular binding site (e.g., compete with members of the transcription factor T-cytokine/lymphokine (TCF/LEF) family at the TCF site on β -catenin). In some embodiments, the provided technology competes with TCF for interaction with β -catenin. In some embodiments, the provided agent binds to a β -catenin site with reduced, inhibited, and/or blocked binding of β -catenin by another binding partner (e.g., a kinase). In some embodiments, binding of the provided agents blocks binding of the TCF/LEF family member to β -catenin. In some embodiments, the present disclosure provides agents that selectively bind to the site of β -catenin over one or more other binding sites of other ligands (e.g., peptides, proteins, etc., in some embodiments, the ligand is Axin, in some embodiments, the ligand is Bcl 9). In some embodiments, provided techniques modulate one or more β -catenin functions associated with its interaction with TCF. In some embodiments, the provided technology selectively modulates β -catenin function, such as a function associated with TCF interactions. In some embodiments, the provided technology selectively modulates β -catenin function and does not significantly affect functions not associated with β -catenin (e.g., multiple functions and/or processes in the Wnt pathway not associated with β -catenin). In some embodiments, the provided techniques may be used to inhibit β -catenin function. In some embodiments, the provided techniques may be used to promote and/or enhance immune activity, such as anti-tumor adaptive immunity.
In some embodiments, the provided techniques may be used to prevent or treat a variety of conditions, disorders, or diseases, including cancer. In some embodiments, the present disclosure provides methods for treating or preventing a condition, disorder, or disease associated with β -catenin comprising administering to a subject suffering from or susceptible to a condition, disorder, or disease associated with β -catenin an effective amount of the provided agent, or a pharmaceutically acceptable salt thereof. In some embodiments, the condition, disorder or disease and β -catenin are associated with TCF interactions. In some embodiments, the agent (e.g., a stapled peptide) is administered as a pharmaceutical composition. In some embodiments, the present disclosure provides pharmaceutical compositions comprising or delivering a provided agent or pharmaceutically acceptable salt thereof. In some embodiments, the pharmaceutical composition further comprises a lipid. As shown herein, in some embodiments, suitable lipids may facilitate delivery/activity. In some embodiments, the agent is or comprises a peptide. In some embodiments, the agent is or comprises a stapled peptide. In some embodiments, provided agents that bind β -catenin comprise one or more engineered amino acid residues.
In some embodiments, the present disclosure provides an agent that binds to a polypeptide comprising or consisting of: SEQ ID NO. 1 (Uniprot ID P35222), or residues 250 to 450 of SEQ ID NO. 1 or residues 305 to 419 of SEQ ID NO. 1:
Uniprot No.P35222
in some embodiments, the provided agents specifically interact with one or more residues that are or correspond to: residues 305 to 419 of SEQ ID NO. 1. In some embodiments, the provided agents bind to a motif (e.g., a portion of a polypeptide, a domain of a polypeptide, etc.) comprising one or more residues corresponding to: ala305, tyr306, gly307, asn308, gln309, lys312, arg342, lys345, val346, val349, gln375, arg376, gln379, asn380, leu382, trp383, arg386, asn387, asp413, asn415, val416, thr418 and Cys419 of SEQ ID NO. 1. In some embodiments, the provided agents bind to a motif (e.g., a portion of a polypeptide, a domain of a polypeptide, etc.) comprising one or more residues corresponding to: ala305, tyr306, gly307, asn308, gln309, lys312, lys345, val346, val349, gln375, arg376, gln379, asn380, leu382, trp383, arg386, asn387, asp413, asn415, val416, thr418 and Cys419 of SEQ ID NO. 1. In some embodiments, the agent binds to a motif comprising one or more of the following residues within SEQ ID NO: 1: ala305, tyr306, gly307, asn308, gln309, lys312, arg342, lys345, val346, val349, gln375, arg376, gln379, asn380, leu382, trp383, arg386, asn387, asp413, asn415, val416, thr418 and Cys419. In some embodiments, the agent binds to a motif comprising one or more of the following residues within SEQ ID NO: 1: ala305, tyr306, gly307, asn308, gln309, lys312, lys345, val346, val349, gln375, arg376, gln379, asn380, leu382, trp383, arg386, asn387, asp413, asn415, val416, thr418 and Cys419. In some embodiments, the agent binds to a motif comprising one or more of the following residues within SEQ ID NO: 1: ala305, tyr306, gly307, asn308, gln309, lys312, arg342, lys345, val346, val349, gln375, gln379, asn380, leu382, trp383, arg386, asn387, asp413, asn415, val416, thr418 and Cys419. In some embodiments, the agent binds to a motif comprising one or more of the following residues within SEQ ID NO: 1: ala305, tyr306, gly307, asn308, gln309, lys312, lys345, val346, val349, gln379, asn380, leu382, trp383, arg386, asn387, asp413, asn415, val416, thr418 and Cys419. In some embodiments, the provided technology binds to motifs comprising at least 2, 3, 4, 5, or 6 of: g307, K312, K345, W383, N387 and N415. In some embodiments, the provided technology binds to motifs comprising at least 2, 3, 4, 5, 6, or 7 of: g307, K312, K345, W383, N387, D413 and N415. In some embodiments, the provided agents bind specifically to such motifs. In some embodiments, the motif may be referred to as a binding site. In some embodiments, the provided technology selectively binds to Axin binding sites as compared to such binding sites. In some embodiments, the provided technology selectively binds to Bcl9 binding sites as compared to such binding sites. In some embodiments, the provided technology selectively binds to such binding sites over TCF binding sites. In some embodiments, the provided technology binds to such binding sites in the opposite N-to-C direction compared to TCF. In some embodiments, the provided technology does not bind to the Axin binding site of β -catenin. In some embodiments, the provided technology does not bind to the Bcl9 binding site of β -catenin. In some embodiments, the provided technology does not bind to the ICAT binding site of β -catenin. In accordance with the present disclosure, interactions with β -catenin can be assessed using a variety of techniques, such as crystallography, NMR, biochemical assays, and the like.
In some embodiments, provided techniques provide for agents, such as stapled peptides, comprising three staple-like structures within 10 to 20, 10 to 15, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 consecutive amino acid residues. In some embodiments, there are three or more staple-like structures within 10 consecutive amino acid residues. In some embodiments, there are three or more staple-like structures within 11 consecutive amino acid residues. In some embodiments, there are three or more staple-like structures within 12 consecutive amino acid residues. In some embodiments, there are three or more staple-like structures within 13 consecutive amino acid residues. In some embodiments, there are three or more staple-like structures within 14 consecutive amino acid residues. In some embodiments, there are three or more staple-like structures within 15 consecutive amino acid residues. In some embodiments, there are three or more staple-like structures within 16 consecutive amino acid residues. In some embodiments, there are three or more staple-like structures within 17 consecutive amino acid residues. In some embodiments, there are three or more staple-like structures within 18 consecutive amino acid residues. In some embodiments, there are three or more staple-like structures within 19 consecutive amino acid residues. In some embodiments, there are three or more staple-like structures within 20 consecutive amino acid residues. In some embodiments, two staple-like structures are bonded to the same amino acid residue. In some embodiments, two staple-like structures are bonded to the same backbone atom. In some embodiments, two staple-like structures are bonded to the same backbone carbon atom. In some embodiments, two staple-like structures are bonded to an a-carbon atom of an amino acid residue and each independently are bonded to another amino acid residue.
In some embodiments, the first staple structure in the agent (e.g., a staple structure peptide) is bonded to the amino acid residues at positions i and i+3. In some embodiments, there is a second staple-like structure bonded to the amino acid residues at positions i+3 and i+10. In some embodiments, there is a third staple-like structure bonded to the amino acid residues at positions i+9 and i+13. Those skilled in the art understand that i, i+3, i+9, i+10, i+13, etc., as used in the art, are generally used to indicate the relevant position of an amino acid residue. In some embodiments, they may also indicate absolute positions in the agent (e.g., peptide). In some embodiments, i is an integer from 1 to 50 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20). In some embodiments, i is 1. In some embodiments, the fourth staple-like structure is present in the agent, e.g., the stapled peptide.
In some embodiments, there are two amino acid residues between two amino acid residues bonded to the same staple-like structure. Such a staple-like structure may be referred to as an (i, i+3) staple-like structure. Similarly, in some embodiments, there are 3, 4, 5, 6, 7, 8, 9, or 10 amino acid residues between two amino acid residues bonded to the same staple-like structure, and such staple-like structures may be referred to as (i, i+4), (i, i+5), (i, i+6), (i, i+7), (i, i+8), (i, i+9), (i, i+10), or (i, i+11), respectively.
In some embodiments, the agent (e.g., a stapled peptide) comprises (i, i+2) and (i, i+7) staple-like structures. In some embodiments, the agent (e.g., a stapled peptide) comprises (i, i+3) and (i, i+7) staple-like structures. In some embodiments, (i, i+3) and (i, i+7) staple structures are bonded to the same amino acid residue. In some embodiments, (i, i+3) and (i, i+7) staple-like structures are bonded to the same atom. In some embodiments, (i, i+3) and (i, i+7) staple-like structures are bonded to the same alpha carbon atom. For example, in compound I-1, (I, i+3) the staple-like structure is bonded to the amino acid residues at positions 1 and 4, and (I, i+7) the staple-like structure is bonded to the amino acid residues at positions 4 and 11, and both of the staple-like structures are bonded to the α -carbon of the amino acid residue at position 4. In some embodiments, the medicament further comprises a third staple-like structure. In some embodiments, the third staple-like structure is (i, i+4). In some embodiments, the third staple-like structure is (i, i+7). In some embodiments, the third staple-like structure is not bonded to any amino acid residues bonded to the first two staple-like structures. In some embodiments, the medicament further comprises a fourth staple-like structure. In some embodiments, the fourth staple-like structure is (i, i+4). In some embodiments, the fourth staple-like structure is (i, i+7). In some embodiments, the fourth staple-like structure is not bonded to any amino acid residues bonded to the first two staple-like structures. In some embodiments, the fourth staple-like structure is not bonded to any amino acid residues bonded to the first three (first thred) staple-like structures.
In some embodiments, provided agents, e.g., peptide agents, e.g., stapled peptide agents, comprise one or more (e.g., 1, 2, 3, 4, 5, 6, or 7) (in some embodiments, from N to C direction) of:
a first acidic group (e.g., of a first acidic amino acid residue);
a second acidic group (e.g., of a second acidic amino acid residue);
optionally a third acidic group (e.g., of a third acidic amino acid residue);
optionally a hydrophobic group (e.g., a hydrophobic amino acid residue);
a first aromatic group (e.g., of a first aromatic amino acid residue);
a second aromatic group (e.g., of a first aromatic amino acid residue); and
a third aromatic group (e.g., of a third aromatic amino acid residue).
In some embodiments, the pharmaceutical agent comprises first and second acidic groups and first, second, and third aromatic groups. In some embodiments, such agents additionally comprise a third acidic group (e.g., of a third acidic amino acid residue) and/or a hydrophobic group (e.g., of a hydrophobic amino acid residue). In some embodiments, such agents additionally comprise a third acidic group (e.g., of a third acidic amino acid residue) and a hydrophobic group Groups (e.g., of hydrophobic amino acid residues). In some embodiments, the distance between the first acidic group and the second acidic group is about the distance between the acidic groups of two acidic amino acid residues of the peptide motif, wherein the distance between the two acidic amino acid residues is about the distance between the acidic groups of the acidic amino acid residues of the peptide motif (e.g., the second at position n+3 if the first acidic amino acid residue is at position N), wherein the distance between the first acidic group and the third acidic group (if present) is about the distance between the acidic groups of two acidic amino acid residues of the peptide motif, wherein the third at position n+4 if the first acidic amino acid residue is at position N), wherein the distance between the first acidic group and the hydrophobic group (e.g., the first amino acid residue is at position n+7 if the first acidic amino acid residue is at position N), wherein the distance between the first acidic amino acid residue and the hydrophobic group is at about the position n+6 if the first acidic amino acid residue is at the position n+6 if the first amino acid residue is at the position n+4 if the first acidic amino acid residue is at the position N), the distance between the first aromatic group and the second aromatic group is about the distance between the aromatic groups of two aromatic amino acid residues of the peptide motif, wherein there are two amino acid residues between the two aromatic amino acid residues (e.g., if the first aromatic amino acid residue is at position M, the second is at position m+3), and/or the distance between the first aromatic group and the third aromatic group is about the distance between the aromatic groups of two aromatic amino acid residues of the peptide motif, wherein there are three amino acid residues between the two aromatic amino acid residues (e.g., if the first aromatic amino acid residue is at position M, the third is at position m+4). In some embodiments, the first acid The neutral amino acid residue is at position N, the second acidic amino acid residue is at position n+3, and the first, second and third aromatic amino acid residues are at positions n+7, n+10 and n+11, respectively. In some embodiments, the first acidic amino acid residue is at position N, the second acidic amino acid residue is at position n+3, the third acidic amino acid residue is at position n+4, and the first, second, and third aromatic amino acid residues are at positions n+7, n+10, and n+11, respectively. In some embodiments, the first acidic amino acid residue is at position N, the second acidic amino acid residue is at position n+3, the hydrophobic amino acid residue is at position n+6, and the first, second, and third aromatic amino acid residues are at positions n+7, n+10, and n+11, respectively. In some embodiments, the first acidic amino acid residue is at position N, the second acidic amino acid residue is at position n+3, the third acidic amino acid residue is at position n+4, the hydrophobic amino acid residue is at position n+6, and the first, second, and third aromatic amino acid residues are at positions n+7, n+10, and n+11, respectively. In some embodiments, M is n+7. In some embodiments, N is 1 to 7. In some embodiments, N is 1, 2, 3, 4, or 5. In some embodiments, N is 1. In some embodiments, N is 2. In some embodiments, N is 3. In some embodiments, N is 4. In some embodiments, N is 5. In some embodiments, M is 8 to 16. In some embodiments, M is 8. In some embodiments, M is 9. In some embodiments, M is 10. In some embodiments, M is 11. In some embodiments, M is 12. In some embodiments, M is 13. In some embodiments, the peptide motif is an alpha helical motif, wherein each amino acid residue is independently an alpha amino acid residue. In some embodiments, the peptide motif is stapled. In some embodiments, there are two or more staple-like structures in the peptide motif; in some embodiments, there are three; in some embodiments, there are four; in some embodiments, there are four or more. In some embodiments, the peptide motif is or comprises the following: agents described herein (e.g., I-xxxx, wherein xxxx is a number (e.g., I-1, I-10, I-100, I-1000, etc.)). In some embodiments In this case, the first acidic group belongs to X as described herein 2 The second acidic group belongs to X as described herein 5 The third acidic group (if present) belongs to X as described herein 6 Hydrophobic groups (if present) belong to X as described herein 8 The first aromatic group belongs to X as described herein 9 The second aromatic group belongs to X as described herein 12 And/or the third aromatic group belongs to X as described herein 13 . In some embodiments, as described herein, the provided agent is a stapled peptide comprising one or more staple-like structures. In some embodiments, as described herein, the provided agent is a stapled peptide comprising two or more staple-like structures. In some embodiments, as described herein, the provided agent is a stapled peptide comprising three or more staple-like structures. In some embodiments, when contacted with a β -catenin polypeptide, the first acidic group interacts with Lys312 and/or Gly307 or corresponding amino acid residues thereof, the second acidic group interacts with Asn387, trp383 and/or Arg386 or corresponding amino acid residues thereof, the first aromatic group interacts with Lys345 and/or Trp383 or corresponding amino acid residues thereof, the second aromatic group interacts with Trp383 and/or Asn415 or corresponding amino acid residues thereof, and the third aromatic group interacts with gin 379, leu383, val416, asn415 and/or Trp383 or corresponding amino acid residues thereof. In some embodiments, the third acidic group interacts with Asn387, trp383, and/or Arg386, or their corresponding amino acid residues. In some embodiments, the hydrophobic group interacts with Trp383 or its corresponding amino acid residue.
In some embodiments, the present disclosure provides an agent having the structure of formula I:
R N -L P1 -L AA1 -L P2 -L AA2 -L P3 -L AA3 -L P4 -L AA4 -L P5 -L AA5 -L P6 -L AA6 -L P7 -R c ，
I
wherein each variable is independently as described herein.
In some embodiments, the present disclosure provides an agent that is or comprises a peptide comprising:
[X 0 ] p0 X 1 X 2 X 3 X 4 X 8 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 ，
wherein:
p0, p15, p16 and p17 are each independently 0 or 1;
X 0 ，X 1 ，X 2 ，X 3 ，X 4 ，X 5 ，X 6 ，X 7 ，X 8 ，X 9 ，X 10 ，X 11 ，X 12 ，X 13 ,X 14 ，X 15 ，X 16 and X 17 Each independently is an amino acid residue.
In some embodiments, the present disclosure provides an agent that is or comprises a peptide comprising:
[X] p X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 [X] p’ ，
wherein:
p15, p16 and p17 are each independently 0 or 1;
p and p' are each independently 0 to 10;
X，X 1 ，X 2 ，X 3 ，X 4 ，X 5 ，X 6 ，X 7 ，X 8 ，X 9 ，X 10 ，X 11 ，X 12 ，X 13 ，X 14 ，X 15 ，X 16 and X 17 Each independently is an amino acid residue.
In some embodiments, the agent is:
R N -[X]pX 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 [X 14 ] p14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 [X]p’-R C wherein each variable is independently as described herein.
In some embodiments, the agent is or comprises the following:
X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 [X 14 ] p14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 [X 18 ] p18 [X 19 ] p19 [X 20 ] p20 [X 21 ] p21 [X 22 ] p22 [X 23 ] p23 wherein p14, p15, p16, p17, p18, p19, p20, p21, p22, and p23 are each independently 0 or 1, and X 1 ，X 2 ，X 3 ，X 4 ，X 5 ，X 6 ，X 7 ，X 8 ，X 9 ，X 10 ，X 11 ，X 12 ，X 13 ，X 14 ，X 15 ，X 16 ，X 17 ，X 18 ，X 19 ，X 20 ，X 21 ，X 22 And X 23 Each independently is an amino acid residue as described herein.
In some embodiments, such peptides comprise three or more staple-like structures. In some embodiments, such peptides comprise five or more residues suitable for stapling.
In some embodiments, the present disclosure provides an agent, wherein the agent is or comprises a peptide comprising:
[X 0 ] p0 X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 ，
wherein:
p0, p15, p16 and p17 are each independently 0 or 1;
X 0 ，X 1 ，X 2 ，X 3 ，X 4 ，X 5 ，X 6 ，X 7 ，X 8 ，X 9 ，X 10 ，X 11 ，X 12 ，X 13 ，X 14 ，X 15 ，X 16 and X 17 Each independently is an amino acid residue, wherein:
X 2 comprising a side chain containing an acidic or polar group;
X 5 comprising a side chain containing an acidic or polar group;
X 13 comprising a side chain containing an optionally substituted aromatic group; and is also provided with
X 1 ，X 3 ，X 4 ，X 7 ，X 10 ，X 11 And X 14 Each independently is an amino acid residue suitable for stapling, or each independently is stapling.
In some embodiments, the present disclosure provides an agent, wherein the agent is or comprises a peptide comprising:
[X 0 ] p0 X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 ，
wherein:
p0, p15, p16 and p17 are each independently 0 or 1;
X 0 ，X 1 ，X 2 ，X 3 ，X 4 ，X 5 ，X 6 ，X 7 ，X 8 ，X 9 ，X 10 ，X 11 ，X 12 ，X 13 ，X 14 ，X 15 ，X 16 and X 17 Each independently is an amino acid residue, wherein:
X 2 comprising a side chain containing an acidic or polar group;
X 5 comprising a side chain containing an acidic or polar group;
X 6 comprising a side chain containing an acidic or polar group;
X 13 comprising a side chain containing an optionally substituted aromatic group; and is also provided with
X 1 ，X 3 ，X 4 ，X 7 ，X 10 ，X 11 And X 14 Each independently is an amino acid residue suitable for stapling, or each independently is stapling.
In some embodiments, the agent is or comprises a peptide. In some embodiments, the agent is or comprises a stapled peptide. In some embodiments, the agent is a peptide. In some embodiments, the agent is a stapled peptide. In some embodiments, the agent, peptide, or stapled peptide has [ X ] 0 ] p0 X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 Is a structure of (a). In some embodiments, X 1 And X 4 And/or X 4 And X 11 Independently are amino acid residues suitable for stapling or are stapled, or X 3 And X 10 Independently are amino acid residues suitable for stapling or are stapled. In some embodimentsWherein X is 1 And X 4 Independently are amino acid residues suitable for stapling. In some embodiments, X 1 And X 4 Is stapled. In some embodiments, X 4 And X 11 Independently are amino acid residues suitable for stapling. In some embodiments, X 4 And X 11 Is stapled. In some embodiments, X 1 And X 4 And X 4 And X 11 Independently are amino acid residues suitable for stapling. In some embodiments, the stapled peptide is a stitched peptide (stitched peptide) comprising two or more staple-like structures, some of which may be bonded to the same backbone atom. In some embodiments, X 1 And X 4 Is stapled, and X 4 And X 11 Is stapled. In some embodiments, linkage X 1 And X 4 Staple-like structure of (c) and connection X 4 And X 11 Staple-like structure of (c) and X 4 Is bonded to the common backbone atom of (c). In some embodiments, the common backbone atom is X 4 Is a carbon alpha to the carbon number. In some embodiments, X 3 And X 10 Independently are amino acid residues suitable for stapling. In some embodiments, X 3 And X 10 Is stapled. In some embodiments, X 1 And X 3 Independently are amino acid residues suitable for stapling. In some embodiments, X 1 And X 3 Is stapled. In some embodiments, X 10 And X 14 Independently are amino acid residues suitable for stapling. In some embodiments, X 10 And X 14 Is stapled. In some embodiments, X 7 And X 10 Independently are amino acid residues suitable for stapling. In some embodiments, X 7 And X 10 Is stapled. In some embodiments, X 7 And X 14 Independently are amino acid residues suitable for stapling. In some embodiments, X 7 And X 14 Is stapled. In some embodiments, X 3 And X 7 Independently are amino acid residues suitable for stapling. In some embodiments, X 3 And X 7 Is stapled.
In some embodiments, the present disclosure provides agents that bind to polypeptides comprising residues 305 to 419 of SEQ ID No. 1 or consisting of residues 305 to 419 of SEQ ID No. 1 as described herein. In some embodiments, the agent (e.g., peptide) has a molecular weight of no more than about 5000 daltons. In some embodiments, it does not exceed about 2500, 3000, 3500, 4000, 4500, or 5000 daltons. In some embodiments, it does not exceed about 2500 daltons. In some embodiments, it does not exceed about 3000 daltons. In some embodiments, it does not exceed about 3500 daltons. In some embodiments, it does not exceed about 4000 daltons. In some embodiments, it does not exceed about 500 daltons.
In some embodiments, the present disclosure provides various techniques, e.g., reagent methods, etc., for preparing, characterizing, evaluating, and using the provided agents and compositions thereof. In some embodiments, the present disclosure provides methods, reagents, and/or systems, e.g., for identifying, characterizing, and/or evaluating provided agents and uses thereof (e.g., as therapeutic or diagnostic agents).
In some embodiments, the present disclosure provides pharmaceutical compositions comprising or delivering a provided agent and a pharmaceutically acceptable carrier. In some embodiments, the provided agent is in the form of a pharmaceutically acceptable salt. In some embodiments, provided compositions comprise a pharmaceutically acceptable salt form of the agent. In some embodiments, in various compositions and methods, the agent is provided as a pharmaceutically acceptable salt form.
In some embodiments, the present disclosure provides methods for modulating a property, activity, and/or function of β -catenin comprising contacting β -catenin with a provided agent. In some embodiments, the present disclosure provides methods for modulating a property, activity, and/or function of β -catenin in a system comprising β -catenin comprising administering to the system an effective amount of the provided agent. In some embodiments, the present disclosure provides methods for modulating a property, activity, and/or function of β -catenin in a system expressing β -catenin comprising administering or delivering an effective amount of the provided agent to the system. In some embodiments, the activity of β -catenin is inhibited or reduced. In some embodiments, the function of β -catenin is inhibited or reduced. In some embodiments, the property, activity, and/or function is associated with β -catenin/TCF interactions.
In some embodiments, the present disclosure provides methods for modulating β -catenin/TCF interactions. In some embodiments, the present disclosure provides methods for modulating β -catenin/TCF interactions comprising contacting β -catenin with a provided agent. In some embodiments, the present disclosure provides methods for modulating β -catenin/TCF interactions in a system comprising β -catenin and TCF, comprising administering or delivering an effective amount of the provided agents to the system. In some embodiments, the present disclosure provides methods for modulating β -catenin/TCF interactions in a system expressing β -catenin and TCF comprising administering or delivering an effective amount of the provided agents to the system. In some embodiments, the interaction between β -catenin and TCF is reduced. In some embodiments, the interaction between β -catenin and TCF is inhibited.
In some embodiments, the present disclosure provides methods for inhibiting cell proliferation comprising administering or delivering an effective amount of the provided agents to a population of cells. In some embodiments, the present disclosure provides methods for inhibiting cell proliferation in a system comprising administering or delivering to the system an effective amount of the provided agent. In some embodiments, the present disclosure provides methods for inhibiting cell growth comprising administering or delivering an effective amount of the provided agents to a population of cells. In some embodiments, the present disclosure provides methods for inhibiting cell growth in a system comprising administering or delivering to the system an effective amount of the provided agent. In some embodiments, such cell proliferation is β -catenin dependent. In some embodiments, such cell growth is β -catenin dependent. In some embodiments, such proliferation or growth is dependent on the interaction of β -catenin with TCF.
In some embodiments, the present disclosure provides methods for reducing or preventing WNT pathway activation. In some embodiments, the present disclosure provides methods for reducing or preventing WNT pathway activation in a system, comprising administering or delivering an effective amount of the provided agent to the system.
In some embodiments, the system is in vitro. In some embodiments, the system is ex vivo. In some embodiments, the system is in vivo. In some embodiments, the system is or comprises a cell. In some embodiments, the system is or comprises an organization. In some embodiments, the system is or comprises an organ. In some embodiments, the system is or comprises an organism. In some embodiments, the system is an animal. In some embodiments, the system is a human. In some embodiments, the system is or comprises a cell, tissue or organ associated with a condition, disorder or disease. In some embodiments, the system is or comprises a cancer cell.
In some embodiments, the present disclosure provides methods for preventing a condition, disorder, or disease. In some embodiments, the present disclosure provides methods for reducing the risk of a condition, disorder, or disease. In some embodiments, the present disclosure provides methods for preventing a condition, disorder, or disease comprising administering or delivering an effective amount of an agent of the present disclosure to a subject susceptible to the condition, disorder, or disease. In some embodiments, the present disclosure provides methods for reducing the risk of a condition, disorder, or disease comprising administering or delivering an effective amount of an agent of the present disclosure to a subject susceptible to the condition, disorder, or disease. In some embodiments, the present disclosure provides methods for reducing the risk of a condition, disorder, or disease in a population comprising administering or delivering an effective amount of an agent of the present disclosure to a population of subjects susceptible to the condition, disorder, or disease. In some embodiments, the present disclosure provides methods for treating a condition, disorder, or disease. In some embodiments, the present disclosure provides methods for treating a condition, disorder, or disease comprising administering or delivering an effective amount of an agent of the present disclosure to a subject suffering from the condition, disorder, or disease. In some embodiments, the symptoms are reduced, eliminated, or prevented. In some embodiments, one or more parameters used to evaluate the condition, disorder, or disease are improved. In some embodiments, survival of the subject is prolonged. As will be appreciated by those of skill in the art, in some embodiments, prophylactic, risk-reducing, and/or therapeutic effects may be assessed by clinical trials and may be observed in a population of subjects. In some embodiments, the condition, disorder or disease is cancer. In some embodiments, the condition, disorder or disease is associated with β -catenin. In some embodiments, the condition, disorder or disease and the interaction of β -catenin with TCF are related. In some embodiments, the condition, disorder or disease is bladder cancer. In some embodiments, the condition, disorder or disease is endometrial cancer. In some embodiments, the condition, disorder or disease is adrenocortical carcinoma. In some embodiments, the condition, disorder or disease is gastric cancer. In some embodiments, the condition, disorder or disease is lung cancer. In some embodiments, the condition, disorder or disease is melanoma. In some embodiments, the condition, disorder or disease is esophageal cancer. In some embodiments, the condition, disorder or disease is colorectal cancer. In some embodiments, the cancer is liver cancer. In some embodiments, the cancer is prostate cancer. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is endometrial cancer. Mutations that lead to constitutive activation of Wnt/β -catenin mediated signaling are reported to be present in about 20% of all human cancers. In some embodiments, the condition, disorder or disease is associated with WNT signaling. In some embodiments, the condition, disorder or disease is associated with β -catenin dependent WNT signaling. In some embodiments, the condition, disorder or disease is associated with β -catenin/TCF interactions. In some embodiments, β -catenin/TCF interactions have been reported to promote cell proliferation, epithelial-mesenchymal transition (EMT), cancer stem cell phenotype, and the like.
In some embodiments, the agent is administered as a pharmaceutical composition comprising or delivering such agent. In some embodiments, the agent is provided and/or delivered in the form of a pharmaceutically acceptable salt. In some embodiments, in a composition (e.g., a liquid composition of a particular pH), the agent may be present in a variety of forms, including a variety of pharmaceutically acceptable salt forms.
In some embodiments, the provided agent is used in combination with a second treatment. In some embodiments, the provided agent is used in combination with a second therapeutic agent. In some embodiments, the second therapeutic or therapeutic agent is administered prior to administration or delivery of the provided agent. In some embodiments, the second therapeutic or therapeutic agent is administered about simultaneously with administration or delivery of the provided agent. In some embodiments, the second therapeutic or therapeutic agent is administered after administration or delivery of the provided agent. In some embodiments, the subject is exposed to both the provided agent and the second therapeutic agent. In some embodiments, the subject is exposed to the therapeutic effect of the provided agent and the therapeutic effect of the second therapeutic agent. In some embodiments, the second treatment is or comprises surgery. In some embodiments, the second treatment is or comprises radiation therapy. In some embodiments, the second treatment is or comprises an immunotherapy. In some embodiments, the second therapeutic agent is or comprises a drug. In some embodiments, the second therapeutic agent is or comprises a cancer drug. In some embodiments, the second therapeutic agent is or comprises a chemotherapeutic agent. In some embodiments, the second therapeutic agent is or comprises a hormonal therapeutic agent. In some embodiments, the second therapeutic agent is or comprises a kinase inhibitor. In some embodiments, the second therapeutic agent is or comprises a checkpoint inhibitor (e.g., an antibody to PD-1, PD-L1, CTLA-4, etc.). In some embodiments, the provided agents may be administered in lower unit doses and/or total doses than used alone. In some embodiments, the second agent may be administered in a lower unit dose and/or total dose than used alone. In some embodiments, one or more side effects associated with administration of the provided agent and/or the second therapeutic or therapeutic agent are reduced. In some embodiments, for example, the combination therapy provides improved results when compared to each agent used alone. In some embodiments, for example, the combination therapy achieves one or more better results when compared to each agent used alone.
Further description of certain implementations of the provided technology is presented below.
Drawings
FIG. 1 provides techniques for selectively inhibiting beta-catenin driven gene transcription in cells expressing beta-catenin. The stapled peptide inhibited endogenous gene expression in wild HAP1 isogenic cells, but did not inhibit endogenous gene expression in CTNNB1 Knockout (KO) cells. (A): beta-catenin levels. CHIR: CHIR99021, which activates the β -catenin pathway and increases AXIN2 and SP5 expression. (B): SP5 expression (24 hours). (C): AXIN2 expression (24 hours). For each group, DMSO ("0" and "0"), peptide A (1 and 5. Mu.M), I-66 (1 and 5. Mu.M), and I-470 (1 and 5. Mu.M) were used from left to right. Expression assessed 24 hours after treatment.
FIG. 2 provides techniques for reducing nuclear β -catenin levels. The results (24 hours) of total β -catenin in the nuclear fraction are shown as an example.
FIG. 3 provides techniques for inhibiting cell proliferation, modulating transcription, and/or inducing cell cycle arrest. (A): the provided technology can reduce cell proliferation. (B) and (C): the provided technology can regulate gene expression. (B): AXIN was used for 24 hours. (C): CXCL12 hours. (D): the provided techniques may induce cell cycle arrest. From left to right: peptides A (1, 5 and 10. Mu.M), I-66 (1, 5 and 10. Mu.M), I-470 (1, 5 and 10. Mu.M) and DMSO.
Fig. 4 the provided technology can provide robust, dose-dependent anti-tumor effects in vivo. Both dose levels evaluated provided a robust reduction in tumor size, and higher dose levels provided a greater reduction. COLO320DM cells (colon carcinoma, mutation: APC, TP 53) were used for the presented data. The top line is the vehicle treatment, the middle line is I-66, 30mg/kg, Q4D, and the bottom line is I-66, 75mg/kg, Q4D.
Fig. 5 provides techniques that can provide sustained tumor exposure, suitable pharmacokinetic profiles, and broad tissue distribution. (A): sustained COLO320DM xenograft tumor exposure following a single i.p. injection of I-66 at 50mg/kg is shown as an example. The dotted line indicates in vitro proliferation IC 50 (0.7. Mu.M). (B): plasma pharmacokinetics in mice. The data presented are as example I-66 plasma concentrations (ng/mL) over time. (C): tissue distribution of I-66 observed in one evaluation. Mice were given a single dose of IP,50mg/kg. For each sample, the left column is 24 hours data and the right column is 96 hours data.
FIG. 6I-66 formulation prepared as described in example 9 1 H NMR(DMSO-d6，373K)。
FIG. 7I-66 formulation prepared as described in example 9 1 Integration of the peaks in the H NMR spectrum (DMSO-d 6, 373K). Those skilled in the art will appreciate that the integration may be further tuned and/or optimized.
Fig. 8 the provided techniques may provide in vivo robust anti-tumor effects in a variety of tumor models. (A): certain data from PDX colon cancer models. (B): some data from the PDX CRC model.
Detailed Description
Definition of the definition
Unless otherwise indicated, the following definitions shall apply. For purposes of this disclosure, CAS versions, handbook of Chemistry and Physics,75, according to the periodic table of elements th Ed to determine the chemical element. In addition, general principles of organic chemistry are described in "Organic Chemistry", thomas Sorrell, university Science Books, sausalito:1999, and "March's Advanced Organic Chemistry”,5 th Ed.,Ed.:Smith,M.B.and March,J.,John Wiley&Sons,New York:2001。
And (3) application: the term "administering" as used herein generally refers to the administration of a composition to a subject or system. Those of ordinary skill in the art will recognize a variety of routes that may be used to administer to a subject (e.g., a human) where appropriate. For example, in some embodiments, administration may be ocular, oral, parenteral, topical, and the like. In some embodiments, administration may be bronchial (e.g., by bronchial instillation), buccal, transdermal (which may be or contain, for example, one or more of surface to dermis, intradermal (inter-dermal), transdermal, etc.), enteral, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, indoor (intra-ocular), specific organ (e.g., intrahepatic), mucosal, nasal, oral, rectal, subcutaneous, sublingual, surface, tracheal (e.g., by intratracheal instillation), vaginal, vitreous, etc. In some embodiments, administration may involve administration as intermittent administration (e.g., multiple doses separated in time) and/or periodic administration (e.g., separate doses separated by a common period). In some embodiments, administration may involve continuous administration (e.g., infusion) for at least a selected period of time.
Affinity: as known in the art, "affinity" is a measure of compactness in the case of binding of a particular ligand (e.g., an agent) to its partner (e.g., β -catenin or a portion thereof). Affinity can be measured in different ways. In some embodiments, the affinity is measured by a quantitative assay. In some such embodiments, the binding partner concentration may be immobilized beyond the ligand concentration to mimic physiological conditions. Alternatively or additionally, in some embodiments, the binding partner concentration and/or ligand concentration may vary. In some such embodiments, the affinity can be compared to a reference under comparable conditions (e.g., concentration).
Medicament: in general, the term "agent" as used herein may be used to refer to a compound or entity of any chemical class, including, for example, polypeptides, nucleic acids, sugars, lipids, small molecules, metals, or combinations or complexes thereof. Where appropriate, the term may be used to refer to or comprise an entity that is a cell or organism or a fraction, extract or component thereof, as will be clear to a person skilled in the art from the context. Alternatively or additionally, as will be clear from the context, the term may be used to refer to a natural product as it is found in and/or obtained from nature. In some cases, again as will be clear from the context, the term may be used to refer to one or more artificial entities, as designed, altered, and/or created by the action of a human hand, and/or not present in nature. In some embodiments, the agent may be used in isolated form or in pure form; in some embodiments, the pharmaceutical agents may be used in crude form. In some embodiments, potential agents may be provided as a collection or library, which may be screened, for example, to identify or characterize active agents therein. In some cases, the term "agent" may refer to a compound or entity that is or comprises a polymer; in some cases, the term may refer to a compound or entity that includes one or more polymeric moieties. In some embodiments, the term "agent" may refer to a compound or entity that is not a polymer and/or is substantially free of any polymer and/or free of one or more specific polymer moieties. In some embodiments, the term may refer to a compound or entity that lacks or is substantially free of any polymeric moiety. In some embodiments, the agent is a compound. In some embodiments, the agent is a stapled peptide.
Aliphatic: as used herein, "aliphatic" means a straight (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is fully saturated or contains one or more unsaturated units, or a substituted or unsubstituted monocyclic, bicyclic, or polycyclic hydrocarbon ring that is fully saturated or contains one or more unsaturated units (but is not aromatic), or a combination thereof. In some embodiments, the aliphatic group comprises 1 to 50 aliphatic carbon atoms. In some embodiments, the aliphatic group comprises 1 to 20 aliphatic carbon atoms. In other embodiments, the aliphatic group comprises 1 to 10 aliphatic carbon atoms. In other embodiments, the aliphatic group comprises 1 to 9 aliphatic carbon atoms. In other embodiments, the aliphatic group comprises 1 to 8 aliphatic carbon atoms. In other embodiments, the aliphatic group comprises 1 to 7 aliphatic carbon atoms. In other embodiments, the aliphatic group comprises 1 to 6 aliphatic carbon atoms. In other embodiments, the aliphatic group comprises 1 to 5 aliphatic carbon atoms, and in other embodiments, the aliphatic group comprises 1, 2, 3, or 4 aliphatic carbon atoms. Suitable aliphatic groups include, but are not limited to, straight or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl, and mixtures thereof (hybrid), such as (cycloalkyl) alkyl, (cycloalkenyl) alkyl or (cycloalkyl) alkenyl.
Alkenyl: the term "alkenyl" as used herein refers to an aliphatic group as defined herein having one or more double bonds.
Alkyl: the term "alkyl" as used herein is given its ordinary meaning in the art and may include saturated aliphatic groups including straight chain alkyl, branched chain alkyl, cycloalkyl (alicyclic) groups, alkyl substituted cycloalkyl, and cycloalkyl substituted alkyl. In some embodiments, the alkyl groups have 1 to 100 carbon atoms. In certain embodiments, the linear or branched alkyl groups have about 1 to 20 carbon atoms in their backbone (e.g., C for a linear chain 1 -C 20 For branched chains C 2 -C 20 ) And alternatively, about 1 to 10. In some embodiments, cycloalkyl rings have about 3 to 10 carbon atoms in their ring structure, wherein such rings are monocyclic, bicyclic, or polycyclic, and instead have about 5, 6, or 7 carbons in the ring structure. In some embodiments, the alkyl group may be a lower alkyl group, wherein the lower alkyl group contains 1 to 4 carbon atoms (e.g., for a straight chain lower alkyl group, C 1 -C 4 )。
Amino groupAcid: as used herein, in its broadest sense refers to any compound and/or substance that can be incorporated into a polypeptide chain, for example, by forming one or more peptide bonds. In some embodiments, the amino acid comprises an amino group and a carboxylic acid group. In some embodiments, the amino acid has NH (R a1 )-L a1 -C(R a2 )(R a3 )-L a2 -COOH structure, wherein each variable is independently as described in the present disclosure. In some embodiments, the amino acid has the general structure NH (R ') -C (R') 2 -COOH, wherein each R' is independently as described in the present disclosure. In some embodiments, the amino acid has the general structure H 2 N-C(R’) 2 -COOH, wherein R' is as described in the disclosure. In some embodiments, the amino acid has the general structure H 2 N-C (H) (R ') -COOH, wherein R' is as described in the present disclosure. In some embodiments, the amino acid is a naturally occurring amino acid. In some embodiments, the amino acid is a non-natural amino acid; in some embodiments, the amino acid is a D-amino acid; in some embodiments, the amino acid is an L-amino acid. "Standard amino acid" refers to any of the twenty standard L-amino acids typically found in naturally occurring peptides. "non-standard amino acid" refers to any amino acid other than a standard amino acid, whether synthetically prepared or obtained from natural sources. In some embodiments, the amino acids (including carboxyl and/or amino terminal amino acids) in the polypeptide may comprise structural modifications as compared to the general structures above. For example, in some embodiments, amino acids can be modified by methylation, amidation, acetylation, pegylation, glycosylation, phosphorylation, and/or substitution of (e.g., amino, carboxylic acid group, one or more protons, one or more hydrogens, and/or hydroxyl groups) as compared to the general structure. In some embodiments, such modifications may, for example, alter the circulating half-life of a polypeptide comprising the modified amino acid as compared to a polypeptide comprising an otherwise identical unmodified amino acid. In some embodiments, such modifications do not significantly alter the inclusion of the modified amino acid as compared to a polypeptide comprising an otherwise identical unmodified amino acid Is a polypeptide of the genus, or a polypeptide of the genus. As will be clear from the context, in some embodiments, the term "amino acid" may be used to refer to a free amino acid; in some embodiments, it may be used to refer to an amino acid residue of a polypeptide.
An analog: the term "analog" as used herein refers to a substance that shares one or more specific structural features, elements, components or portions with the reference substance. In general, "analogs" exhibit significant structural similarity to a reference substance, such as a shared core or shared structure (consensus structure), but also differ in some discrete ways. In some embodiments, the analog is a substance that can be generated from a reference substance, for example, by chemically manipulating the reference substance. In some embodiments, an analog is a substance that can be produced by performing a synthetic process that is substantially similar (e.g., shares multiple steps with) a synthetic process that produces a reference substance. In some embodiments, the analog is produced by or can be produced by performing a different synthetic process than that used to produce the reference substance.
Animals: as used herein, refers to any member of the animal kingdom. In some embodiments, "animal" refers to a human of either sex and at any stage of development. In some embodiments, "animal" refers to a non-human animal at any stage of development. In certain embodiments, the non-human animal is a mammal (e.g., rodent, mouse, rat, rabbit, monkey, dog, cat, sheep, cow, primate, and/or pig). In some embodiments, animals include, but are not limited to, mammals, birds, reptiles, amphibians, fish, insects, and/or worms. In some embodiments, the animal may be a transgenic animal, a genetically engineered animal, and/or a clone.
About: the term "about" or "approximately," as used herein, when applied to one or more destination values, refers to values similar to the reference value. In certain embodiments, unless otherwise specified or otherwise evident from the context (unless such numbers exceed 100% of the possible values), the term "about" or "approximately" refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less of any direction (greater than or less) of the reference value.
Aryl: the term "aryl" used alone or as part of a larger moiety such as "aralkyl", "aralkoxy", "aryloxyalkyl", and the like, refers to a monocyclic, bicyclic, or polycyclic ring system having a total of 5 to 30 ring members, wherein at least one ring in the system is aromatic. In some embodiments, aryl is a monocyclic, bicyclic, or polycyclic ring system having a total of 5 to 14 ring members, wherein at least one ring in the system is aromatic, and wherein each ring in the system comprises 3 to 7 ring members. In some embodiments, the aryl group is a biaryl group. The term "aryl" may be used interchangeably with the term "aryl ring". In certain embodiments of the present disclosure, "aryl" refers to an aromatic ring system including, but not limited to, phenyl, biphenyl, naphthyl, binaphthyl, anthracenyl, and the like, which may carry one or more substituents. Also included within the scope of the term "aryl" as used herein are groups in which an aromatic ring is fused to one or more non-aromatic rings, such as indanyl, phthalimidyl (phtalimidyl), naphthalimidyl (napthoimidyl), phenanthridinyl, tetrahydronaphthyl, or the like, wherein the linking group or point of attachment is on the aryl ring.
Associated with … …: as the term is used herein, an event or entity is "related to" one another if the presence, level, and/or form of the event or entity is related to the presence, level, and/or form of another event or entity. For example, a particular entity (e.g., nucleic acid (e.g., genomic DNA, transcript, mRNA, etc.), polypeptide, genetic marker (genetic signature), metabolite, microorganism, etc.) is considered to be associated with a particular disease, disorder, or condition if its presence, level, and/or form is associated with the occurrence and/or susceptibility of the disease, disorder, or condition (e.g., in an associated population).
Combining: it will be understood that the term "binding" as used herein generally refers to non-covalent association between or within agents. In many embodiments herein, binding involves binding to β -catenin to a particular agent. Those of ordinary skill in the art will appreciate that such binding may be evaluated in any of a variety of situations. In some embodiments, binding is assessed against β -catenin. In some embodiments, binding is assessed against one or more amino acid residues of β -catenin. In some embodiments, the binding is evaluated for one or more amino acid residues corresponding to those of β -catenin (e.g., similarly positioned in three-dimensional space and/or having certain similar properties and/or functions).
Binding sites: the term "binding site" as used herein refers to a region of a target polypeptide formed in three dimensions that comprises one or more or all of the interacting residues of the target polypeptide. In some embodiments, a "binding site" may refer to one or more amino acid residues comprising or being one or more or all of the interacting amino acid residues of a target polypeptide. As will be appreciated by one of ordinary skill in the art, the binding sites may include residues that are linear adjacent to each other and/or residues that are linear distal to each other but proximal to each other in three dimensions when the target polypeptide is folded. The binding site may comprise amino acid residues and/or sugar residues.
And (3) a carrier: as used herein, refers to a diluent, adjuvant, excipient, or carrier with which the composition is administered. In some exemplary embodiments, the carrier may comprise a sterile liquid, such as, for example, water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as, for example, peanut oil, soybean oil, mineral oil, sesame oil and the like. In some embodiments, the carrier is or includes one or more solid components.
The method is equivalent to that of: the term "comparable" as used herein refers to two or more agents, entities, conditions, sets of conditions, etc., which may not be identical to each other but are similar enough to allow comparison between them, and thus, one of skill in the art will understand that conclusions can be drawn reasonably based on the observed differences or similarities. In some embodiments, a group of comparable conditions, environments, individuals, or populations is characterized by a plurality of substantially identical features and one or a small number of varying features. In this context, one of ordinary skill in the art will understand what degree of identity is required for two or more such agents, entities, situations, condition sets, etc. in any given instance is considered equivalent. For example, one of ordinary skill in the art will appreciate that the environment, individual, or group of populations are equivalent to one another when: characterized by having a sufficient number and type of substantially identical features to ensure a reasonable conclusion-differences in the results or observed phenomena obtained under different circumstances, groups or circumstances of individuals or populations are caused by or indicative of the changes in the features of these changes.
Composition: those skilled in the art will appreciate that the term "composition" may be used to refer to a discrete physical entity comprising one or more specified components. In general, unless otherwise indicated, the composition may have any form-e.g., gas, gel, liquid, solid, etc.
Cycloaliphatic: the term "cycloaliphatic" as used herein refers to a saturated or partially unsaturated aliphatic mono-, bi-, or polycyclic ring system having, for example, 3 to 30 members, wherein the aliphatic ring system is optionally substituted. Cycloaliphatic groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, cyclooctyl, cyclooctenyl, norbornyl (norbornyl), adamantyl (adamantyl), and cyclooctadienyl. In some embodiments, cycloalkyl groups have 3 to 6 carbons. The term "cycloaliphatic" may also include aliphatic rings, such as decalin or tetrahydrodecalin, fused to one or more aromatic or non-aromatic rings wherein the linking group or point of attachment is on the aliphatic ring. In some embodiments, the carbocyclic group is bicyclic. In one place In some embodiments, the carbocyclic group is tricyclic. In some embodiments, the carbocyclic group is polycyclic. In some embodiments, "cycloaliphatic" (or "carbocycle" or "cycloalkyl") refers to a monocyclic C 3 -C 10 Or C 3 -C 6 Hydrocarbons or C 4 -C 10 Or C 8 -C 10 Bicyclic hydrocarbons which are fully saturated or contain one or more unsaturated units but which are not aromatic, or C 9 -C 16 Tricyclic hydrocarbons, which are fully saturated or contain one or more unsaturated units but which are not aromatic.
Derivatives: the term "derivative" as used herein refers to a structural analogue of a reference substance. That is, "derivatives" are substances that exhibit significant structural similarity with a reference substance, such as a shared core or shared structure, but also differ in some discrete fashion. In some embodiments, the derivative is a substance that can be produced from a reference substance by a chemical procedure. In some embodiments, a derivative is a substance that can be produced by performing a synthetic process that is substantially similar (e.g., shares multiple steps with) a synthetic process that produces a reference substance.
Dosage form or unit dosage form: those skilled in the art will appreciate that the term "dosage form" may be used to refer to physically discrete units of an active agent (e.g., a therapeutic agent or diagnostic agent) for administration to a subject. Typically, each such unit contains a predetermined amount of active agent. In some embodiments, such amount is a unit dose amount (or an entire fraction thereof) suitable for administration according to a dosing regimen determined to be associated with a desired or beneficial outcome when administered to the relevant population (i.e., according to a therapeutic dosing regimen). Those of ordinary skill in the art will appreciate that the total amount of therapeutic composition or agent administered to a particular subject is determined by one or more attending physicians and may involve the administration of multiple dosage forms.
Dosing regimen: those skilled in the art will appreciate that the term "dosing regimen" may be used to refer to groups of unit doses (typically more than one) that are administered to a subject individually, typically at intervals over a period of time. In some embodiments, a given therapeutic agent has a recommended dosing regimen that may involve one or more doses. In some embodiments, the dosing regimen comprises a plurality of doses, each dose being spaced apart in time from the other doses. In some embodiments, the individual doses are spaced apart from each other by a period of the same length; in some embodiments, the dosing regimen comprises a plurality of doses and at least two different periods of time separating the individual doses. In some embodiments, all doses within a dosing regimen have the same unit dose amount. In some embodiments, different doses within a dosing regimen have different amounts. In some embodiments, the dosing regimen comprises a first dose in a first dose amount followed by one or more additional doses in a second dose amount different from the first dose amount. In some embodiments, the dosing regimen comprises a first dose in a first dose amount followed by one or more additional doses in a second dose amount identical to the first dose amount. In some embodiments, the dosing regimen is associated with a desired or beneficial outcome (i.e., it is a therapeutic dosing regimen) when administered in the relevant population.
And (3) modifying: in general, the term "engineered" refers to an aspect that has been manipulated by a person. For example, in some embodiments, a peptide may be considered engineered if the amino acid sequence of the peptide is selected by a human. For example, the engineered agent has an amino acid sequence selected based on the preference for the corresponding amino acid at a particular site of protein-protein interaction. In some embodiments, the engineered sequence has an amino acid sequence that is different from the amino acid sequence of a polypeptide contained in the NCBI database that binds to the TCF site of β -catenin. In many embodiments, the provided agent is an engineered agent. In some embodiments, the engineered agent is a peptide agent comprising an unnatural amino acid residue, an unnatural amino acid sequence, and/or a peptide staple-like structure. In some embodiments, the provided agents comprise or are engineered peptide agents comprising an engineered sequence.
Halogen: the term "halogen" means F, cl, br or I.
Heteroaliphatic: the term "heteroaliphatic" is given its ordinary meaning in the art and refers to an aliphatic group as described herein wherein one or more carbon atoms are replaced with one or more heteroatoms (e.g., oxygen, nitrogen, sulfur, silicon, phosphorus, etc.).
Heteroalkyl: the term "heteroalkyl" is given its ordinary meaning in the art and refers to an alkyl as described herein wherein one or more carbon atoms are replaced with a heteroatom (e.g., oxygen, nitrogen, sulfur, silicon, phosphorus, etc.). Some examples of heteroalkyl groups include, but are not limited to, alkoxy, poly (ethylene glycol) -, alkyl-substituted amino, tetrahydrofuranyl, piperidinyl, morpholinyl, and the like.
Heteroaryl group: the term "heteroaryl" and variations thereof, as used alone or as part of a larger moiety (e.g., "heteroaralkyl" or "heteroaralkoxy"), refers to a monocyclic, bicyclic, or polycyclic ring system having, for example, a total of 5 to 30 (e.g., 5, 6, 9, 10, 14, etc.) ring members, wherein at least one ring in the system is aromatic and at least one aromatic ring atom is a heteroatom. In some embodiments, the heteroatom is nitrogen, oxygen, or sulfur. In some embodiments, heteroaryl is a group having 5 to 10 ring atoms (i.e., monocyclic, bicyclic, or polycyclic), in some embodiments 5, 6, 9, or 10 ring atoms. In some embodiments, heteroaryl groups have 6, 10, or 14 pi electrons that are common in a cyclic array (cyclic array); and has 1 to 5 heteroatoms in addition to carbon atoms. Heteroaryl groups include, but are not limited to: thienyl, furyl pyrrole group, imidazole group pyrazolyl, triazolyl, tetrazolyl, Azolyl, iso->Azolyl, (-) -and (II) radicals>Diazolyl (oxadiazyl), thiazolyl, isothiazolyl, thiadiazolyl, and the like,Pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl (indolizinyl), purinyl, naphthyridinyl and pteridinyl. In some embodiments, the heteroaryl is a heteroaryl, such as bipyridyl, and the like. The term "heteroaryl" and variations thereof as used herein also include groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclyl rings, wherein the linking group or point of attachment is on the heteroaromatic ring. Some non-limiting examples include: indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzothiazolyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido [2,3-b ]]-1,4-Oxazin-3 (4H) -one. Heteroaryl groups may be monocyclic, bicyclic or polycyclic. The term "heteroaryl" may be used interchangeably with the terms "heteroaryl ring", "heteroaryl group" or "heteroaromatic", any of which include an optionally substituted ring. The term "heteroarylalkyl" refers to an alkyl group substituted with a heteroaryl group, wherein the alkyl and heteroaryl moieties are independently optionally substituted.
Heteroatom(s): the term "heteroatom" means an atom that is not carbon and is not hydrogen. In some embodiments, the heteroatom is oxygen, sulfur, nitrogen, phosphorus, boron or silicon (including any oxidized form of nitrogen, sulfur, phosphorus or silicon; quaternized forms of any basic or substitutable nitrogen of a heterocycle (e.g., N (as in 3, 4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or NR + (as in N-substituted pyrrolidinyl); etc.). In some embodiments, the heteroatom is boron, nitrogen, oxygen, silicon, sulfur, or phosphorus. In some embodiments, the heteroatom is nitrogen, oxygen, silicon, sulfur, or phosphorus. In some embodiments, the heteroatom is nitrogen, oxygen, sulfur, or phosphorus. In some embodiments, the heteroatom is nitrogen, oxygen, or sulfur.
A heterocyclic group: as used hereinThe terms "heterocycle", "heterocyclyl", "heterocyclic group" and "heterocyclic ring" are used interchangeably and refer to a monocyclic, bicyclic or polycyclic ring portion (e.g., 3 to 30 membered) that is saturated or partially unsaturated and has one or more heteroatom ring atoms. In some embodiments, the heteroatom is boron, nitrogen, oxygen, silicon, sulfur, or phosphorus. In some embodiments, the heteroatom is nitrogen, oxygen, silicon, sulfur, or phosphorus. In some embodiments, the heteroatom is nitrogen, oxygen, sulfur, or phosphorus. In some embodiments, the heteroatom is nitrogen, oxygen, or sulfur. In some embodiments, the heterocyclyl is a stable 5-to 7-membered monocyclic or 7-to 10-membered bicyclic heterocyclic moiety which is saturated or partially unsaturated and has one or more, preferably one to four heteroatoms as defined above in addition to carbon atoms. When used with respect to a ring atom of a heterocycle, the term "nitrogen" includes substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 0 to 3 heteroatoms selected from oxygen, sulfur or nitrogen, the nitrogen may be N (as in 3, 4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or + NR (as in N-substituted pyrrolidinyl). The heterocyclic ring may be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure, and any ring atom may be optionally substituted. Some examples of such saturated or partially unsaturated heterocyclic groups include, but are not limited to: tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl,Oxazolidinyl, piperazinyl, dioxanyl, diaza +.>Radical (diazepinyl), oxazal->Radical (oxazepinyl), thiazate +.>A group (thiazepinyl), a morpholinyl group and a quinuclidinyl group. The terms "heterocycle", "heterocyclyl ring", "heterocyclic group (heterocyclic group)", "heterocyclic moiety" and "heterocyclic group (heterocyclic radical)" are used interchangeably herein and also include groups in which the heterocyclyl ring is fused to one or more aryl, heteroaryl or cycloaliphatic rings, such as indolinyl, 3H-indolyl, chromanyl (chromanyl), phenanthridinyl or tetrahydroquinolinyl, in which the linking group or point of attachment is on the cycloaliphatic ring. The heterocyclyl groups may be monocyclic, bicyclic or polycyclic. The term "heterocyclylalkyl" refers to an alkyl group substituted with a heterocyclyl group, where the alkyl and heterocyclyl moieties are independently optionally substituted.
Homology: the term "homology" as used herein refers to the overall relatedness between polymer molecules, e.g. between nucleic acid molecules (e.g. DNA molecules and/or RNA molecules) and/or between polypeptide molecules. In some embodiments, the sequences of the polymer molecules are considered "homologous" to each other if they have at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 99% identity. In some embodiments, the sequences of the polymer molecules are considered "homologous" to each other if they have at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% similarity (e.g., contain residues with related chemical properties at the corresponding positions). For example, certain amino acids are generally classified as similar to each other as "hydrophobic" or "hydrophilic" amino acids, and/or as having "polar" or "nonpolar" side chains, as known to those of ordinary skill in the art. Substitution of one amino acid for another amino acid of the same type can generally be considered a "homologous" substitution. Typical amino acid classifications are summarized below (Kyte and Doolittle, hydrophobicity scale of 1982:A simple method for displaying the hydropathic character of a protein.J.Mol.Biol.157:105-132):
Polysense amino acid (Ambiguous Amino Acid) 3 letters 1 letter
Asparagine or aspartic acid Asx B
Glutamine or glutamic acid Glx Z
Leucine or isoleucine Xle J
Unspecified or unknown amino acids Xaa X
As will be appreciated by those skilled in the art, there are a variety of algorithms available that allow sequences to be compared to determine their degree of homology, including by allowing one sequence to have gaps of a specified length relative to another when considering which residues in the different sequences "correspond" to each other. For example, the calculation of the percent homology between two nucleic acid sequences can be performed by aligning the two sequences for optimal comparison purposes (e.g., gaps can be introduced in one or both of the first and second nucleic acid sequences for optimal alignment, and non-corresponding sequences can be ignored for comparison purposes). In certain embodiments, the length of the sequences aligned for comparison purposes is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or substantially 100% of the length of the reference sequence. The nucleotides at the corresponding nucleotide positions are then compared. When a position in the first sequence is occupied by the same nucleotide as the corresponding position in the second sequence, then the molecules at that position are identical; when a position in the first sequence is occupied by a nucleotide that is similar to the corresponding position in the second sequence, then the molecules at that position are similar. The percent homology between two sequences is a function of the number of identical and similar positions shared by the sequences, which needs to be introduced to achieve optimal alignment of the two sequences, taking into account the number of gaps and the length of each gap. Representative algorithms and computer programs that can be used to determine the percent homology between two nucleotide sequences include, for example, the algorithm of Meyers and Miller (CABIOS, 1989, 4:11-17) which has been incorporated into the ALIGN program (version 2.0) using the PAM120 weight residue table with a gap length penalty of 12 and a gap penalty of 4. Alternatively, the percent homology between two nucleotide sequences may be determined using, for example, the nwsgapdna.cmp matrix using the GAP program in the GCG software package.
Interaction residues: the terms "interacting residues", "interacting motif" as used herein refer to residues or motifs in an agent that are designed to interact with a particular target residue in a target polypeptide, or residues in a target polypeptide that interact with a particular motif (e.g., aromatic group, amino acid residue, etc.) of an agent, in the case of a target polypeptide. In particular, the interaction residues and motifs of the plurality of agents are selected and arranged within the agents such that they will be displayed in three-dimensional space (e.g., upon binding, docking, or other interaction measurement) within a predetermined distance (or volume) of the determined target residues. In many embodiments, the interaction residue is a direct binding residue.
"improve", "increase" or "decrease": these terms or grammatically equivalent comparative terms as used herein indicate measured values relative to equivalent references. For example, in some embodiments, an evaluation value achieved with a drug of interest may be "improved" relative to an evaluation value obtained with a comparable reference drug. Alternatively or additionally, in some embodiments, the evaluation value achieved in a subject or system of interest may be "improved" relative to an evaluation value obtained under different conditions (e.g., before or after an event such as administration of the agent of interest) in the same subject or system, or in a different equivalent subject (e.g., in the presence of one or more indicators of a particular disease, disorder, or condition of interest, or in an equivalent subject or system that is different from the subject or system of interest in the event of prior exposure to the condition or agent, etc.). In some embodiments, the comparative term refers to a statistically relevant difference (e.g., having a prevalence and/or number sufficient to achieve a statistical correlation). In a given context, those skilled in the art will recognize or will be able to readily determine the degree and/or prevalence of the difference required or sufficient to achieve such statistical significance.
Partially unsaturated: the term "partially unsaturated" as used herein refers to a moiety that contains at least one double or triple bond. The term "partially unsaturated" is intended to encompass groups having multiple sites of unsaturation, but is not intended to encompass aryl or heteroaryl moieties.
Peptide: the term "peptide" as used herein refers to a polypeptide. In some embodiments, the peptide is a relatively short polypeptide, e.g., less than about 100 amino acids, less than about 50 amino acids, less than about 40 amino acids, less than about 30 amino acids, less than about 25 amino acids, less than about 20 amino acids, less than about 15 amino acids, or less than 10 amino acids in length. In some embodiments, the length is about 5 to 20, 5 to 19, 5 to 18, 5 to 17, 5 to 16, 5 to 15, 10 to 20, 10 to 19, 10 to 18, 10 to 17, 10 to 16, 10 to 15, 11 to 20, 11 to 19, 11 to 18, 11 to 17, 11 to 16, 11 to 15, 12 to 20, 12 to 19, 12 to 18, 12 to 17, 12 to 16, 12 to 15, 13 to 20, 13 to 19, 13 to 18, 13 to 17, 13 to 16, 13 to 15, 14 to 20, 14 to 19, 14 to 18, 14 to 17, 14 to 16, 14 to 15, or about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids.
Pharmaceutical composition: the term "pharmaceutical composition" as used herein refers to an active agent formulated with one or more pharmaceutically acceptable carriers. In some embodiments, the active agent is present in a unit dose amount suitable for administration in a treatment regimen that, when administered to a relevant population, exhibits a statistically significant probability of achieving a predetermined therapeutic effect. In some embodiments, the pharmaceutical compositions may be specifically formulated for administration in solid or liquid form, including those suitable for: oral administration, such as drench (aqueous or non-aqueous solutions or suspensions), tablets (e.g., those for buccal, sublingual and systemic absorption), boluses (boluses), powders, granules, pastes for application to the tongue; parenteral administration, for example, by subcutaneous, intramuscular, intravenous or epidural injection as, for example, a sterile solution or suspension, or a sustained release formulation; topical application, for example as a cream, ointment or controlled release patch or spray applied to the skin, lungs or oral cavity; intravaginal or intrarectal, for example as pessaries (pessary), creams or foams; sublingual; ocular menstruation; percutaneous; or nasally, pulmonary, and to other mucosal surfaces.
The medicine can be used for: the phrase "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
Pharmaceutically acceptable carrier: the term "pharmaceutically acceptable carrier" as used herein means a pharmaceutically acceptable material, composition or carrier, such as a liquid or solid filler, diluent, excipient or solvent encapsulating material, involved in carrying or transporting the subject compound from one organ or portion of the body to another organ or portion of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injuring the patient. Some examples of materials that may be used as pharmaceutically acceptable carriers include: sugars such as lactose, glucose, and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerol, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; non-thermal raw water; isotonic saline; ringeR's solution; ethanol; a pH buffer solution; polyesters, polycarbonates and/or polyanhydrides; and other non-toxic compatible substances used in pharmaceutical formulations.
A pharmaceutically acceptable salt: the term "pharmaceutically acceptable salt" as used herein refers to salts of such compounds which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known. For example, pharmaceutically acceptable salts are described in detail in J.pharmaceutical Sciences,66:1-19 (1977) by S.M. Berge et al. In some embodiments, pharmaceutically acceptable salts include, but are not limited to, non-toxic acid addition salts, which are salts of amino groups formed with inorganic acids (e.g., hydrochloric, hydrobromic, phosphoric, sulfuric, and perchloric acids) or with organic acids (e.g., acetic, maleic, tartaric, citric, succinic, or malonic acids) or by using other known methods (e.g., ion exchange). In some embodiments, pharmaceutically acceptable salts include, but are not limited to, adipates, alginates,Ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodite, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectate (pectate), persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. In some embodiments, pharmaceutically acceptable salts include, but are not limited to, non-toxic base addition salts, such as those formed from the acidic groups of the provided compounds with a base. Representative alkali or alkaline earth metal salts include salts of sodium, lithium, potassium, calcium, magnesium, and the like. In some embodiments, the pharmaceutically acceptable salt is an ammonium salt (e.g., -N (R)) 3 + ). In some embodiments, the pharmaceutically acceptable salt is a sodium salt. In some embodiments, pharmaceutically acceptable salts include nontoxic ammonium, quaternary ammonium, and amine cations formed using counter ions (e.g., halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, alkyl, sulfonate, and arylsulfonate groups having 1 to 6 carbon atoms), as appropriate.
Polypeptide: as used herein refers to any polymer chain of amino acids. In some embodiments, the polypeptide has an amino acid sequence that occurs in nature. In some embodiments, the polypeptide has an amino acid sequence that does not exist in nature. In some embodiments, the polypeptide has an engineered amino acid sequence in that it is designed and/or produced by the action of a human hand. In some embodiments, the polypeptide may comprise, or consist of, a natural amino acid, an unnatural amino acid, or both. In some embodiments, the polypeptide may comprise or consist of only natural amino acids or only unnatural amino acids. In some embodiments, the polypeptide may comprise a D-amino acid, an L-amino acid, or both. In some embodiments, the polypeptide may comprise only D-amino acids. In some embodiments, the polypeptide may comprise only L-amino acids. In some embodiments, the polypeptide may comprise one or more pendant groups or other modifications, such as modifications or attachments to one or more amino acid side chains, at the N-terminus of the polypeptide, the C-terminus of the polypeptide, or any combination thereof. In some embodiments, such pendant groups or modifications may be selected from acetylation, amidation, lipidation, methylation, pegylation, and the like, including combinations thereof. In some embodiments, the polypeptide may be cyclic, and/or may comprise a cyclic moiety. In some embodiments, the polypeptide is not cyclic and/or does not comprise any cyclic moiety. In some embodiments, the polypeptide is linear. In some embodiments, the polypeptide may be or comprise a stapled polypeptide. In some embodiments, the term "polypeptide" may be appended to the name of a reference polypeptide, activity, or structure; in such cases, it is used herein to refer to polypeptides that share a related activity or structure, and thus may be considered members of the same class or family of polypeptides. For each such class, the present description provides and/or those skilled in the art will recognize exemplary polypeptides within the class whose amino acid sequence and/or function is known; in some embodiments, such exemplary polypeptides are reference polypeptides for a class or family of polypeptides. In some embodiments, members of a polypeptide class or family exhibit significant sequence homology or identity to a reference polypeptide of that class, share a common sequence motif (e.g., a characteristic sequence element) with a reference polypeptide of that class, and/or share a common activity (in some embodiments at a comparable level or within a specified range) with a reference polypeptide of that class; in some embodiments, with all polypeptides within the class). For example, in some embodiments, the member polypeptide exhibits a degree of sequence homology or identity to the reference polypeptide of at least about 30 to 40%, and typically greater than about 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more, and/or comprises at least one region exhibiting very high sequence identity (typically greater than 90% or even 95%, 96%, 97%, 98% or 99%), for example, may be a conserved region of a characteristic sequence element or a conserved region comprising a characteristic sequence element in some embodiments. Such conserved regions typically cover at least 3 to 4, and typically up to 20 or more amino acids; in some embodiments, the conserved region encompasses at least one segment of at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more contiguous amino acids. In some embodiments, the related polypeptide may comprise or consist of a fragment of the parent polypeptide. In some embodiments, a useful polypeptide may comprise or consist of a plurality of fragments, each of which is present in the same parent polypeptide in a different spatial arrangement relative to each other present in the polypeptide of interest (e.g., directly linked fragments in a parent may be spatially separated in the polypeptide of interest or vice versa, and/or fragments may be present in the polypeptide of interest in a different order than in the parent), such that the polypeptide of interest is a derivative of its parent polypeptide.
Prevention (pre) or prophylaxis (pre): as used herein, when used in connection with the occurrence of a disease, disorder, and/or condition, refers to reducing the risk of developing the disease, disorder, and/or condition and/or delaying the onset of one or more features or symptoms of the disease, disorder, or condition. Prevention may be considered complete when the onset of the disease, disorder or condition is delayed for a predetermined period of time.
Protecting group: the term "protecting group" as used herein is well known in the art and includes those described in detail below: protecting Groups in Organic Synthesis, T.W.Greene and P.G.M.Wuts,3 rd edition,John Wiley&Sons,1999, incorporated herein by reference in its entirety. Also included are those protecting groups described in the following that are particularly suitable for nucleoside and nucleotide chemistry: current Protocols in Nucleic Acid Chemistry, (edited by Serge L.Beaucage et al.) 06/2012, the entirety of chapter 2 of which is incorporated herein by reference. Suitable ammoniaThe radical protecting group comprises methyl carbamate, ethyl carbamate, 9-fluorenylmethyl carbamate (9-fluorenylmethyl carbamate, fmoc), 9- (2-sulfo) fluorenylmethyl carbamate, 9- (2, 7-dibromo) fluorenylmethyl carbamate, 2, 7-di-tert-butyl- [9- (10, 10-dioxo-10, 10-tetrahydrothioxanthyl) ]Methyl carbamate (DBD-Tmoc), 4-methoxybenzoyl methyl carbamate (Phenoc), 2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl carbamate (Teoc), 2-phenylethyl carbamate (hZ), 1- (1-adamantyl) -1-methylethyl carbamate (Adpoc), 1-dimethyl-2-haloethyl carbamate 1, 1-dimethyl-2, 2-dibromoethylcarbamate (DB-t-BOC), 1-dimethyl-2, 2-Trichloroethylcarbamate (TCBOC), 1-methyl-1- (4-biphenylyl) ethylcarbamate (Bpoc), 1- (3, 5-di-tert-butylphenyl) -1-methylethylcarbamate (t-Bumeoc), 2- (2 '-and 4' -pyridyl) ethylcarbamates (Pyoc), 2- (N, N-dicyclohexylformamide) ethylcarbamate, t-Butylcarbamate (BOC), 1-adamantylcarbamate (Adoc), vinylcarbamate (Voc), allylcarbamate (Alloc), 1-isopropylallylcarbamate (IPaoc), cinnamylcarbamate (Coc), 4-Nitrocinnamyl carbamate (Noc), 8-quinolinyl carbamate, N-hydroxypiperidinyl carbamate, alkyldithioyl carbamate, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz), p-nitrobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzyl carbamate, 2, 4-dichlorobenzyl carbamate, 4-methylsulfinylbenzyl carbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl carbamate, 2-methylsulfinylethyl carbamate, 2- (p-toluenesulfonyl) ethyl carbamate, [2- (1, 3-dithianyl) ]Methyl carbamate (Dmoc), 4-methylthiophenyl carbamate (Mtpc), 2, 4-dimethylthiophenyl carbamate (Bmpc), 2-phosphoethyl carbamate (Peoc), 2-triphenylphosphine isopropyl carbamate (Ppoc), 1-dimethyl-2-cyanoethyl carbamate, m-chloro-p-acyloxybenzyl carbamate,P- (dihydroxyboron) benzyl carbamate, 5-benzisoAzolylmethylcarbamates, 2- (trifluoromethyl) -6-colour ketomethylcarbamates (Tcroc), m-nitrophenylcarbamates, 3, 5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate, 3, 4-dimethoxy-6-nitrobenzyl carbamate, phenyl (o-nitrophenyl) methylcarbamate, phenothiazinyl- (10) -carbonyl derivative, N ' -p-toluenesulfonylaminocarbonyl derivative, N ' -phenylthiocarbonyl derivative, t-amyl carbamate, S-benzylthiocarbamate, p-cyanobenzyl carbamate, cyclobutylcarbamate, cyclohexylcarbamate, cyclopentyl carbamate, cyclopropylmethylcarbamate, p-decyloxybenzyl carbamate, 2-dimethoxycarbonyl vinylcarbamate, o- (N, N-dimethylformamide) benzyl carbamate, 1-dimethyl-3- (N, N-dimethylformamide) propyl carbamate, 1-dimethylpropynyl carbamate, di (2-pyridyl) methyl carbamate, 2-furylmethylcarbamate, 2-iodoethylcarbamate, isozhi-alkylcarbamate (isoborynl carbamate), isobutyl carbamate, isonicotinyl carbamate, p- (p ' -methoxyphenylazo) benzyl carbamate, 1-methylcyclobutyl carbamate, 1-methylcyclohexyl carbamate, 1-methyl-1-cyclopropylmethyl carbamate, 1-methyl-1- (3, 5-dimethoxyphenyl) ethylcarbamate, 1-methyl-1- (p-phenylazophenyl) ethylcarbamate, 1-methyl-1-phenylethylcarbamate, 1-methyl-1- (4-pyridyl) ethylcarbamate, phenylcarbamate, p- (phenylazo) benzyl carbamate, 2,4, 6-tri-tert-butylphenylcarbamate, 4- (trimethylammonium) benzyl carbamate, 2,4, 6-trimethylbenzyl carbamate, formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3-phenylpropionamide, picolinamide, 3-pyridylmethylamide, N-benzoylphenylpropionamide derivatives, benzamide, p-phenylbenzene Formamide, o-nitrophenylacetamide, o-nitrophenoxyacetamide, acetoacetamide, (N' -dithiobenzyloxycarbonylamino) acetamide, 3- (p-hydroxyphenyl) propionamide, 3- (o-nitrophenyl) propionamide, 2-methyl-2- (o-nitrophenoxy) propionamide, 2-methyl-2- (o-phenylazophenoxy) propionamide, 4-chlorobutyramide, 3-methyl-3-nitrobutyramide, o-nitrocinnamamide, N-acetylmethionine derivatives, o-nitrobenzamide, o- (benzoyloxymethyl) benzamide, 4, 5-diphenyl-3->Oxazolin-2-one, N-phthalimide, N-dithiosuccinimide (Dts), N-2, 3-diphenylmaleimide, N-2, 5-dimethylpyrrole, N-1, 4-tetramethyldisilylazalane adduct (STABASE), 5-substituted 1, 3-dimethyl-1, 3, 5-triazacyclohexane-2-one, 5-substituted 1, 3-dibenzyl-1, 3, 5-triazacyclohexane-2-one, 1-substituted 3, 5-dinitro-4-pyridone, N-methylamine, N-allylamine, N- [2- (trimethylsilyl) ethoxy]Methylamine (SEM), N-3-acetoxypropylamine, N- (1-isopropyl-4-nitro-2-oxo-3-pyrrolin-3-yl) amine, quaternary ammonium salt, N-benzylamine, N-bis (4-methoxyphenyl) methylamine, N-5-dibenzocycloheptylamine, N-triphenylmethylamine (Tr), N- [ (4-methoxyphenyl) diphenylmethyl ]Amine (MMTr), N-9-phenylfluorenylamine (PhF), N-2, 7-dichloro-9-fluorenylmethylamine, N-ferrocenylmethylamine (Fcm), N-2-pyridylmethylamino N' -oxide, N-1, 1-dimethylthiomethyleneamine, N-benzylidene amine, N-p-methoxybenzylidene amine, N-diphenylmethyleneamine, N- [ (2-pyridyl) isopropylidene]Methyleneamine, N- (N ', N ' -dimethylaminomethylene) amine, N ' -isopropylidenediamine, N-p-nitrobenzyleneamine, N-salicylideneamine, N-5-chlorosalicyleneamine, N- (5-chloro-2-hydroxyphenyl) phenylmethyleneamine, N-cyclohexylamine, N- (5, 5-dimethyl-3-oxo-1-cyclohexenyl) amine, N-borane derivatives, N-diphenylboronic acid derivatives, N- [ phenyl (pentacarbonylchromium or tungsten) carbonyl]Amine, N-copper chelate, N-zinc chelate, N-nitrosamine, amine N-oxide, diphenylphosphinamide (diphenylphosphinamide, D)pp), dimethylthiophosphinamide (Mpt), diphenylthiophosphinamide (Ppt), dialkylphosphoramidate (dialkyl phosphoramidate), dibenzylaminophosphate, diphenylphosphoramidate, phenylsulfenamide (benzenesulfenamide), o-nitrobenzenesulfinamide (Nps), 2, 4-dinitrobenzene sulfinamide, pentachlorobenzene sulfinamide, 2-nitro-4-methoxybenzene sulfinamide, triphenylmethyl sulfinamide, 3-nitropyridine sulfinamide (Npys), p-toluenesulfonamide (Ts), benzenesulfonamide, 2,3,6, -trimethyl-4-methoxybenzene sulfonamide (Mtr), 2,4, 6-trimethoxybenzene sulfonamide (Mtb), 2, 6-dimethyl-4-methoxybenzene sulfonamide (Pme), 2,3,5, 6-tetramethyl-4-methoxybenzene sulfonamide (Mte), 4-methoxybenzene sulfonamide (Mbs), 2,4, 6-trimethylbenzene sulfonamide (Mts), 2, 6-dimethoxy benzene sulfonamide (Mts), trimethyl-4-methoxybenzene sulfonamide (Mbs), trimethyl-8, 6-trimethyl benzene sulfonamide (Mts), trimethyl-4-methoxy benzene sulfonamide (Mbs), trimethyl-4, 6-methoxy benzene sulfonamide (Mbs), trimethyl-4-methoxy benzene sulfonamide (Mbs), and 5, 8' -trimethyl-benzene sulfonamide (Mbs).
In some embodiments, suitable mono-protected amines include, but are not limited to, aralkylamines, carbamates, allylamines, amides, and the like. Some examples of suitable mono-protected amino moieties include t-butoxycarbonylamino (-NHBOC), ethoxycarbonylamino, methoxycarbonylamino, trichloroethoxycarbonylamino, allyloxycarbonylamino (-NHAlloc), benzyloxycarbonylamino (-NHCBZ), allylamino, benzylamino (-NHBn), fluorenylmethylcarbonyl (-NHFmoc), formylamino, acetamido, chloroacetamido, dichloroacetamido, trichloroacetamido, phenylacetylamino, trifluoroacetamido, benzoylamino, t-butyldiphenylsilyl, and the like. In some embodiments, suitable di-protected amines include amines substituted with two substituents independently selected from those described above as mono-protected amines, and also include cyclic imides, such as phthalimide, maleimide, succinimide, and the like. In some embodiments, suitable di-protected amines include pyrrole and the like, 2, 5-tetramethyl- [1,2,5] azadisilazane (2, 5-tetramethyl- [1,2,5] azadis ilidine) and the like, and azide.
Suitable protected carboxylic acids also include, but are not limited to silyl-, alkyl-, alkenyl-, aryl-, and arylalkyl-protected carboxylic acids. Some examples of suitable silyl groups include trimethylsilyl, triethylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, triisopropylsilyl, and the like. Some examples of suitable alkyl groups include methyl, benzyl, p-methoxybenzyl, 3, 4-dimethoxybenzyl, trityl, t-butyl, tetrahydropyran-2-yl. Some examples of suitable alkenyl groups include allyl. Some examples of suitable aryl groups include optionally substituted phenyl, biphenyl, or naphthyl. Some examples of suitable aralkyl groups include optionally substituted benzyl (e.g., p-methoxybenzyl (MPM), 3, 4-dimethoxybenzyl, o-nitrobenzyl, p-halobenzyl, 2, 6-dichlorobenzyl, p-cyanobenzyl), and 2-and 4-picolyl. In some embodiments, suitable protected carboxylic acids include, but are not limited to, optionally substituted C 1-6 Aliphatic esters, optionally substituted aryl esters, silyl esters, activated esters, amides, hydrazides, and the like. Some examples of such ester groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, benzyl, and phenyl, wherein each group is optionally substituted. Additional suitable protected carboxylic acids include Oxazolines and orthoesters.
Suitable hydroxy protecting groups include methyl, methoxymethyl (MOM), methylthiomethyl (methylthiomethyl, MTM), t-butylthiomethyl, (phenyldimethylsilyl) methoxymethyl (SMOM), benzyloxymethyl (bezoyloxymethyl, BOM), p-methoxybenzyloxymethyl (p-methoxybenzyloxymethyl, PMBM), (4-methoxyphenoxy) methyl (p-AOM), guaiacolmethyl (GUM), t-butoxymethyl, 4-Pentenoxymethyl (POM), siloxymethyl, 2-methoxyethoxymethyl (MEM), 2-trichloroethoxymethyl, bis (2-chloroethoxy) methyl, 2- (trimethylsilyl) ethoxymethyl (semo), tetrahydropyranyl (THP), 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl (MTM), 4-methoxytetrahydropyranyl, 4-methoxypyranyl, 4-methoxythiopyranyl, 3-S-methoxymethyl, 3-tetrahydropyranyl, 3-S-methoxymethyl, 3-chloro-7-methyl, 7-2-methoxyethoxy-methyl, 7-p-m, 3-tetrahydrofuranyl, 7-1-methoxyethoxy-m, 7-methyl, 7-3-dimethyl-1-p-ethyl-7-n-7-methyl-7-3-methoxyethoxy-m, 7-ethyl-7-n-7-3-methyl-7-n-3-methoxyethoxy-m-yl-2-n-methyl-n-2-methoxyethoxymethyl, 1- (2-chloroethoxy) ethyl, 1-methyl-1-methoxyethyl, 1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl, 2-trichloroethyl, 2-trimethylsilylethyl, 2- (phenylselenyl) ethyl (2- (phenylselenyl) ethyl), tert-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2, 4-dinitrophenyl, benzyl, p-methoxybenzyl, 3, 4-dimethoxybenzyl, o-nitrobenzyl, p-halobenzyl, 2, 6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picolyl N-oxide, diphenylmethyl, p, p '-dinitrobenzhydryl (p, p' -dinitabenzhydryl), 5-dibenzocycloheptyl, triphenylmethyl, alpha-naphthyldiphenylmethyl, p-methoxyphenyldiphenylmethyl, di (p-methoxyphenyl) phenylmethyl, tri (p-methoxyphenyl) methyl, 4- (4 '-bromobenzoylmethylphenyl) diphenylmethyl, 4',4 '-tris (4, 5-dichlorophthalimidophenyl) methyl, 4' -tris (levulinyloxyphenyl) methyl (4, 4',4"-tris (levulinoyloxyphenyl) methyl), 4',4" -tris (benzoyloxyphenyl) methyl, 3- (imidazol-1-yl) bis (4 ', 4' -dimethoxyphenyl) methyl, 1-bis (4-methoxyphenyl) -1' -pyrenylmethyl, 9-anthryl, 9- (9-phenyl) xanthenyl, 9- (9-phenyl-10-oxo) anthryl, 1, 3-benzodithiophene-2-yl, S-dioxabenzisothiazolyl, trimethylsilyl (TMS), triethylsilyl (triethylsilyl, TES), triisopropylsilyl (triisopropylsilyl, TIPS), dimethylisopropylsilyl (IPMS), diethylisopropylsilyl (DEIPS), dimethylhexylsilyl (dimethylhexylsilyl), t-butyldimethylsilyl (t-butylsilyl), TBDMS), t-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TBMPS), formate, benzoylformate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxovalerate (levulinate), 4- (ethyldithio) valerate (levulinyl dithioacetal), pivalate, adamantanoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate, 2,4, 6-trimethylbenzoate (mesitoate), alkyl methyl carbonate, 9-fluorenylmethyl carbonate (Fmoc), alkyl ethyl carbonate, alkyl 2, 2-trichloroethyl carbonate (Troc), 2- (trimethylsilyl) ethyl carbonate (2- (trimethylsilyl) ethyl carbonate, TMSEC), 2- (phenylsulfonyl) ethyl carbonate (2- (phenylsulfonyl) ethyl carbonate, psec), 2- (triphenylphosphine) ethyl carbonate (Peoc), alkylisobutyl carbonate, alkylvinyl carbonate, alkylallyl carbonate, alkyl p-nitrophenyl carbonate, alkylbenzyl carbonate, alkyl p-methoxybenzyl carbonate, alkyl 3, 4-dimethoxybenzyl carbonate, alkyl o-nitrobenzyl carbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzylthiocarbonate, 4-ethoxy-1-naphthyl carbonate, methyldithiocarbonate, 2-iodobenzoate, 4-azidobutyrate, 4-nitro-4-methylpentanoate, o- (dibromomethyl) benzoate, 2-formylbenzenesulfonate, 2- (methylthiomethoxy) ethyl, 4- (methylthiomethoxy) butyrate, 2- (methylthiomethoxymethyl) benzoate, 2, 6-dichloro-4-methylphenoxyacetate, 2, 6-dichloro-4- (1, 3-tetramethylbutyl) phenoxyacetate, 2, 4-bis (1, 1-dimethylpropyl) phenoxyacetate, chlorodiphenylacetate, isobutyrate, monosuccinate, (E) -2-methyl-2-butenoate, o- (methoxycarbonyl) benzoate, alpha-naphthoate, nitrate, alkyl N, N, N ', N' -tetramethyl diamide phosphate, alkyl N-phenylcarbamates, borates, dimethylphosphinothioate, alkyl 2, 4-dinitrophenyl sulfenates, sulfates, methanesulfonates (mesylates), benzyl sulfonates, and tosylate (Ts). For protecting 1, 2-or 1, 3-diols, protecting groups include methylene acetal, ethylene acetal, 1-t-butylethylene ketal, 1-phenylethylene ketal, (4-methoxyphenyl) ethylene acetal, 2-trichloroethylene acetal, acetonide, cyclopentylene ketal, cyclohexylene ketal, cycloheptylene ketal, benzylidene acetal, p-methoxybenzylidene acetal, 2, 4-dimethoxybenzylidene ketal, 3, 4-dimethoxybenzylidene acetal, 2-nitrobenzylidene acetal, methoxymethylene acetal, ethoxymethylene acetal, dimethoxymethylene orthoester, 1-methoxyethylene orthoester, 1-ethoxyethylene orthoester, 1, 2-dimethoxyethylene orthoester, α -methoxybenzylidene orthoester, 1- (N, N-dimethylamino) ethylene derivative, α - (N, N' -dimethylamino) benzylidene derivative, 2-oxacyclopentylidene ester, di-t-butylsilylidene (di-t-butylsilylene group), 1, 3-dimethoxymethylene orthoester, 1- (3, 3-tetrabutoxyethylene carbonate), cyclic tetraboronic acid derivative (TBDS), and cyclic tetraboronic acid derivative (TBDS).
In some embodiments, the hydroxy protecting group is acetyl, t-butyl, t-butoxymethyl, methoxymethyl, tetrahydropyranyl, 1-ethoxyethyl, 1- (2-chloroethoxy) ethyl, 2-trimethylsilylethyl, p-chlorophenyl, 2, 4-dinitrophenyl, benzyl, benzoyl, p-phenylbenzoyl, 2, 6-dichlorobenzyl, diphenylmethyl, p-nitrobenzyl, triphenylmethyl (trityl), 4' -dimethoxytrityl, trimethylsilyl, triethylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, triphenylsilyl, triisopropylsilyl, benzoylformate, chloroacetyl, trichloroacetyl, trifluoroacetyl, pivaloyl, 9-fluorenylmethylcarbonate, methanesulfonate, toluenesulfonate, trifluoromethanesulfonate, trityl, monomethoxytrityl (monomthoxyrityl, MMTr), 4' -dimethoxytrityl (DMTr) and 4,4' -trimethoxytrityl (TMTr), 2-cyanoethyl (CE or Cne), 2- (trimethylsilyl) ethyl (TSE), 2- (2-nitrophenyl) ethyl, 2- (4-cyanophenyl) ethyl 2- (4-nitrophenyl) ethyl (NPE), 2- (4-nitrobenzenesulfonyl) ethyl, 3, 5-dichlorophenyl, 2, 4-dimethylphenyl, 2-nitrophenyl, 4-nitrophenyl, 2,4, 6-trimethylphenyl, 2- (2-nitrophenyl) ethyl, ding Liudai carbonyl, 4' -tris (benzoyloxy) trityl, diphenylcarbamoyl, levulinic acid, 2- (dibromomethyl) benzoyl (Dbmb), 2- (isopropylthiomethoxymethyl) benzoyl (Ptmt), 9-phenylxanthen-9-yl (pixyl) or 9- (p-methoxyphenyl) xanthin-9-yl (MOX). In some embodiments, the hydroxyl protecting groups are each independently selected from acetyl, benzyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, and 4,4' -dimethoxytrityl. In some embodiments, the hydroxyl protecting group is selected from the group consisting of trityl, monomethoxytrityl, and 4,4' -dimethoxytrityl. In some embodiments, a phosphorus bond protecting group is a group that is attached to a phosphorus bond (e.g., an internucleotide bond) throughout oligonucleotide synthesis. In some embodiments, the protecting group is attached to the sulfur atom of the phosphorothioate group. In some embodiments, the protecting group is attached to an oxygen atom of the internucleotide phosphorothioate linkage. In some embodiments, the protecting group is attached to an oxygen atom of an internucleotide phosphate linkage. In some embodiments, the protecting group is 2-cyanoethyl (CE or Cne), 2-trimethylsilylethyl, 2-nitroethyl, 2-sulfonylethyl, methyl, benzyl, o-nitrobenzyl, 2- (p-nitrophenyl) ethyl (NPE or NPE), 2-phenylethyl, 3- (N-tert-butylcarboxamido) -1-propyl, 4-oxopentyl, 4-methylsulfanyl-1-butyl, 2-cyano-1, 1-dimethylethyl, 4-N-methylaminobutyl, 3- (2-pyridinyl) -1-propyl, 2- [ N-methyl-N- (2-pyridinyl) ] aminoethyl, 2- (N-formyl, N-methyl) aminoethyl, or 4- [ N-methyl-N- (2, 2-trifluoroacetyl) amino ] butyl.
Protected thiol groups are well known in the art and include those described in detail in Greene (1999). Suitable protected thiol groups also include, but are not limited to, disulfides, sulfides, silyl sulfides, thioesters, thiocarbonates, thiocarbamates, and the like. Some examples of such groups include, but are not limited to, alkyl sulfides, benzyl sulfides and substituted benzyl sulfides, triphenylmethyl sulfides and trichloroethoxycarbonyl thioesters, to name a few.
Reference is made to: as used herein, a standard or control is described with respect to which a comparison is made. For example, in some embodiments, an agent, animal, individual, population, sample, sequence, or value of interest is compared to a reference or control agent, animal, individual, population, sample, sequence, or value. In some embodiments, the reference or control is tested and/or assayed substantially simultaneously with the test or assay of interest. In some embodiments, the reference or control is a historical reference or control, optionally embodied in a tangible medium. Generally, as will be appreciated by those skilled in the art, a reference or control is assayed or characterized under conditions or conditions comparable to those under evaluation. When sufficient similarity exists to demonstrate reliance on and/or comparison with a particular possible reference or control, those skilled in the art will understand.
Specificity: as known in the art, "specificity" is a measure of the ability of a particular ligand (e.g., an agent) to distinguish its binding partner (e.g., β -catenin) from other potential binding partners (e.g., another protein, another portion (e.g., domain) of β -catenin).
Substitution: as described herein, the compounds of the present disclosure may include optionally substituted and/or substituted moieties. In general, the term "substituted", whether preceded by the term "optional", means that one or more hydrogens of the designated moiety are replaced with a suitable substituent. Unless otherwise indicated, an "optionally substituted" group may have suitable substituents at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from the specified group, the substituents may be the same or different at each position. Combinations of substituents contemplated by the present disclosure are preferably those that result in the formation of stable or chemically feasible compounds. The term "stable" as used herein means that the compound does not substantially change when subjected to conditions that allow for its production, detection, and in certain embodiments its recovery, purification, and use for one or more of the purposes disclosed herein. In some embodiments, some exemplary substituents are described below.
Suitable monovalent substituents are halogen; - (CH) 2 ) 0-4 R o ；-(CH 2 ) 0-4 OR o ；-O(CH 2 ) 0-4 R o ，-O-(CH 2 ) 0-4 C(O)OR o ；-(CH 2 ) 0-4 CH(OR o ) 2 The method comprises the steps of carrying out a first treatment on the surface of the Can be R o Substituted- (CH) 2 ) 0-4 Ph; can be R o Substituted- (CH) 2 ) 0-4 O(CH 2 ) 0-1 Ph; can be R o Substituted-ch=chph; can be R o Substituted- (CH) 2 ) 0-4 O(CH 2 ) 0-1 -a pyridinyl group; -NO 2 ；-CN；-N 3 ；-(CH 2 ) 0-4 N(R o ) 2 ；-(CH 2 ) 0-4 N(R o )C(O)R o ；-N(R o )C(S)R o ；-(CH 2 ) 0-4 N(R o )C(O)N(R o ) 2 ；-N(R o )C(S)N(R o ) 2 ；-(CH 2 ) 0-4 N(R o )C(O)OR o ；-N(R o )N(R o )C(O)R o ；-N(R o )N(R o )C(O)N(R o ) 2 ；-N(R o )N(R o )C(O)OR o ；-(CH 2 ) 0-4 C(O)R o ；-C(S)R o ；-(CH 2 ) 2-4 C(O)OR o ；-(CH 2 ) 0-4 C(O)SR o ；-(CH 2 ) 0-4 C(O)OSi(R o ) 3 ；-(CH 2 ) 0-4 OC(O)R o ；-OC(O)(CH ２ ) 0-4 SR o ，-SC(S)SR o ；-(CH 2 ) 0-4 SC(O)R o ；-(CH 2 ) 0-4 C(O)N(R o ) 2 ；-C(S)N(R o ) 2 ；-C(S)SR o ；-SC(S)SR o ，-(CH 2 ) 0-4 OC(O)N(R o ) 2 ；-C(O)N(OR o )R o ；-C(O)C(O)R o ；-C(O)CH 2 C(O)R o ；-C(NOR o )R o ；-(CH 2 ) 0-4 SSR o ；-(CH 2 ) 0-4 S(O) 2 R o ；-(CH 2 ) 0-4 S(O) 2 OR o ；-(CH 2 ) 0-4 OS(O) 2 R o ；-S(O) 2 N(R o ) 2 ；-(CH 2 ) 0-4 S(O)R o ；-N(R o )S(O) 2 N(R o ) 2 ；-N(R o )S(O) 2 R o ；-N(OR o )R o ；-C(NH)N(R o ) 2 ；-Si(R o ) 3 ；-OSi(R o ) 3 ；-P(R o ) 2 ；-P(OR o ) 2 ；-OP(R o ) 2 ；-OP(OR o ) 2 ；-N(R o )P(R o ) 2 ，-B(R o ) 2 ；-OB(R o ) 2 ；-P(O)(R o ) 2 ；-OP(O)(R o ) 2 ；-N(R o )P(O)(R o ) 2 ；-(C 1-4 Linear or branched alkylene) O-N (R) o ) 2 The method comprises the steps of carrying out a first treatment on the surface of the Or- (C) 1-4 Straight or branched chain alkylene) C (O) O-N (R) o ) 2 The method comprises the steps of carrying out a first treatment on the surface of the Wherein each R is o Can be substituted as defined below and is independently hydrogen, C 1-20 Aliphatic, C having 1 to 5 heteroatoms independently selected from nitrogen, oxygen, sulfur, silicon and phosphorus 1-20 Heteroaliphatic, -CH 2 -(C 6-14 Aryl group), -O (CH 2 ) 0-1 (C 6-14 Aryl group, -CH 2 - (5-to 14-membered heteroaryl ring), a 5-to 20-membered monocyclic, bicyclic or polycyclic saturated, partially unsaturated or aryl ring having 0-to 5-heteroatoms independently selected from nitrogen, oxygen, sulfur, silicon and phosphorus, or two independently occurring, although as defined aboveR o Taken together with intervening atoms to form a 5-to 20-membered, monocyclic, bicyclic or polycyclic, saturated, partially unsaturated or aryl ring having 0 to 5 heteroatoms independently selected from nitrogen, oxygen, sulfur, silicon and phosphorus, which may be substituted as defined below.
R o (or by two independently-occurring R' s o Ring formed with intervening atoms) is independently halogen, - (CH) 2 ) 0-2 R · - (halo R) · )、-(CH 2 ) 0-2 OH、-(CH 2 ) 0-2 OR · 、-(CH 2 ) 0-2 CH(OR · ) 2 (halo) R · )、-CN、-N 3 、-(CH 2 ) 0-2 C(O)R · 、-(CH 2 ) 0-2 C(O)OH、-(CH 2 ) 0-2 C(O)OR · 、-(CH 2 ) 0-2 SR · 、-(CH 2 ) 0-2 SH、-(CH 2 ) 0-2 NH 2 、-(CH 2 ) 0-2 NHR · 、-(CH 2 ) 0-2 NR · 2 、-NO 2 、-SiR · 3 、-OSiR · 3 、-C(O)SR · ，-(C 1-4 Straight-chain OR branched alkylene) C (O) OR · or-SSR · Wherein each R is · Is unsubstituted or substituted with one or more halogens only in the case of "halo" preceding, and is independently selected from C 1-4 Aliphatic, -CH 2 Ph，-O(CH 2 ) 0-1 Ph, or a 5-to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur. At R o Suitable divalent substituents on saturated carbon atoms of (c) include =o and =s.
Suitable divalent substituents are the following: =o, =s, =nnr * 2 、＝NNHC(O)R * 、＝NNHC(O)OR * 、＝NNHS(O) 2 R * 、＝NR * 、＝NOR * 、-O(C(R * 2 )) 2-3 O-or-S (C (R) * 2 )) 2-3 S-, each of whichR independently of each other * Selected from hydrogen, C which may be substituted as defined below 1-6 Aliphatic, or unsubstituted 5-to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur. Suitable divalent substituents bonded to the ortho-substitutable carbon of an "optionally substituted" group include: -O (CR) * 2 ) 2-3 O-, wherein each independently occurs R * Selected from hydrogen, C being substituted as defined below 1-6 Aliphatic, or unsubstituted 5-to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.
At R * Suitable substituents on the aliphatic radical of (2) are halogen, -R · - (halo R) · )、-OH、-OR · (halo) R · )、-CN、-C(O)OH、-C(O)OR · 、-NH 2 、-NHR · 、-NR · 2 or-NO 2 Wherein each R is · Unsubstituted or substituted with one or more halogen groups only in the case of "halo" preceding them, and is independently C 1-4 Aliphatic, -CH 2 Ph，-O(CH 2 ) 0-1 Ph, or a 5-to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.
In some embodiments, suitable substituents on the substitutable nitrogen are Each of which is provided withIndependently hydrogen, C which may be substituted as defined below 1-6 Aliphatic, unsubstituted-OPh, or having a structure independently selected from nitrogen, oxygen and sulfurUnsubstituted 5-to 6-membered saturated, partially unsaturated or aryl ring of 0 to 4 heteroatoms, or, in spite of the above definition, two independently occurring->Taken together with intervening atoms form an unsubstituted 3 to 12 membered saturated, partially unsaturated or aryl monocyclic or bicyclic ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur.
At the position ofSuitable substituents on the aliphatic radical of (2) are independently halogen, -R · - (halo R) · )、-OH、-OR · (halo) R · )、-CN、-C(O)OH、-C(O)OR · 、-NH 2 、-NHR · 、-NR · 2 or-NO 2 Wherein each R is · Unsubstituted or substituted with one or more halogen groups only in the case of "halo" preceding them, and is independently C 1-4 Aliphatic, -CH 2 Ph，-O(CH 2 ) 0-1 Ph, or a 5-to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.
The object is: the term "subject" or "subject" as used herein refers to any organism to which provided compounds or compositions are administered in accordance with the present disclosure, e.g., for experimental, diagnostic, prophylactic and/or therapeutic purposes. Typical subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans; insects; worms; and the like) as well as plants. In some embodiments, the subject may have and/or be susceptible to a disease, disorder, and/or condition. In some embodiments, the subject is a human.
Is easy to suffer from: an individual who is "susceptible to" a disease, disorder and/or condition is an individual who is at a higher risk of developing the disease, disorder and/or condition than is a member of the general public. In some embodiments, an individual susceptible to a disease, disorder, and/or condition may not have been diagnosed with the disease, disorder, and/or condition. In some embodiments, an individual susceptible to a disease, disorder, and/or condition may exhibit symptoms of the disease, disorder, and/or condition. In some embodiments, an individual susceptible to a disease, disorder, and/or condition may not exhibit symptoms of the disease, disorder, and/or condition. In some embodiments, an individual susceptible to a disease, disorder, and/or condition will develop the disease, disorder, and/or condition. In some embodiments, an individual susceptible to a disease, disorder, and/or condition will not develop the disease, disorder, and/or condition.
Target polypeptide: the term "target polypeptide" as used herein is a polypeptide with which an agent interacts. In some embodiments, the target polypeptide is a β -catenin polypeptide. In some embodiments, the target polypeptide comprises, consists essentially of, or is a binding site for a β -catenin polypeptide.
Target residues: the term "target residue" as used herein is a residue within a target polypeptide that is designed to interact with an agent. For example, an agent may be characterized by a particular interaction motif (e.g., an aromatic group as described herein) and/or residue (e.g., an amino acid residue comprising an aromatic group as described herein) selected and arranged (by presentation on a selected scaffold) within a certain predetermined distance (or volume) of a target residue. In some embodiments, the target residue is or comprises an amino acid residue.
Therapeutic agent: the phrase "therapeutic agent" as used herein refers to an agent that has a therapeutic effect and/or causes a desired biological and/or pharmacological effect when administered to a subject. In some embodiments, a therapeutic agent is any substance that is useful for alleviating, ameliorating, alleviating, inhibiting, preventing, delaying the onset of, lessening the severity of, and/or reducing the occurrence of one or more symptoms or features thereof.
Treatment protocol: the term "treatment regimen" as used herein refers to a dosing regimen whose administration in the relevant population may be correlated with a desired or beneficial therapeutic outcome.
Therapeutically effective amount of: the term "therapeutically effective amount" as used herein means the amount of a substance (e.g., therapeutic agent, composition, and/or formulation) that elicits the desired biological response when administered as part of a therapeutic regimen. In some embodiments, a therapeutically effective amount of a substance is an amount sufficient to treat, diagnose, prevent, and/or delay the onset of a disease, disorder, and/or condition when administered to a subject suffering from or susceptible to such disease, disorder, and/or condition. As will be appreciated by one of ordinary skill in the art, the effective amount of the substance may vary depending on such factors as the desired biological endpoint, the substance to be delivered, the target cell or tissue, and the like. For example, an effective amount of a compound in a formulation for treating a disease, disorder, and/or condition is an amount that reduces, ameliorates, alleviates, inhibits, prevents, delays onset of, reduces the severity of, and/or reduces the occurrence of one or more symptoms or features thereof. In some embodiments, the therapeutically effective amount is administered in a single dose; in some embodiments, multiple unit doses are required to deliver a therapeutically effective amount.
Treatment: the term "treatment" and variations thereof as used herein refers to any method for partially or completely alleviating, ameliorating, alleviating, inhibiting, preventing, delaying the onset of, lessening the severity of, and/or reducing the occurrence of one or more symptoms or features thereof. The treatment may be administered to a subject that does not exhibit signs of the disease, disorder, and/or condition. In some embodiments, the treatment may be administered to a subject that exhibits only early signs of a disease, disorder, and/or condition, e.g., for the purpose of reducing the risk of developing a pathological condition associated with the disease, disorder, and/or condition.
Unit dose: the expression "unit dose" as used herein refers to an amount administered as a single dose and/or in physically discrete units of a pharmaceutical composition. In many embodiments, the unit dose comprises a predetermined amount of the active agent. In some embodiments, the unit dose comprises an entire single dose of the agent. In some embodiments, more than one unit dose is administered to achieve a total single dose. In some embodiments, it is desirable or contemplated that multiple unit doses be administered in order to achieve the desired effect. The unit dose may be, for example, a volume of a liquid (e.g., an acceptable carrier) containing a predetermined amount of one or more therapeutic agents, a predetermined amount of one or more therapeutic agents in solid form, a sustained release formulation or drug delivery device containing a predetermined amount of one or more therapeutic agents, or the like. It will be appreciated that the unit dose may be present in a formulation that includes any of a variety of components in addition to the therapeutic agent. For example, an acceptable carrier (e.g., a pharmaceutically acceptable carrier), diluent, stabilizer, buffer, preservative, etc., may be included as described below. Those of skill in the art will appreciate that in many embodiments, the total suitable daily dose of a particular therapeutic agent may comprise a fraction of a unit dose or multiple unit doses, and may be determined, for example, by the attending physician within the scope of sound medical judgment. In some embodiments, the specific effective dosage level for any particular subject or organism may depend on a variety of factors, including the disorder being treated and the severity of the disorder; the activity of the particular active compound employed; the specific composition used; age, body weight, general health, sex, and diet of the subject; the time of application and rate of excretion of the particular active compound employed; duration of treatment; medicaments and/or additional treatments used in combination or simultaneously with the particular compounds employed; and similar factors well known in the medical arts.
Unsaturated: the term "unsaturated" as used herein means that a moiety has one or more unsaturated units.
Salts of the provided compounds, such as pharmaceutically acceptable acid or base addition salts, stereoisomeric forms and tautomeric forms, are included unless otherwise indicated.
Unless the context clearly indicates otherwise, (i) nouns without quantitative word modifications as used in this disclosure herein may be understood to mean "at least one"; (ii) the term "or" is understood to mean "and/or"; (iii) The terms "comprising," including, "and" containing "(whether used with or without limitation to" the use of) are to be construed as encompassing the listed components or steps individually or in combination with one or more additional components or steps; (iv) The term "another" may be understood to mean at least another/a second or more; (v) The terms "about" and "approximately" are to be understood as allowing standard variations, as understood by one of ordinary skill in the art; and (vi) where ranges are provided, inclusive of the endpoints.
Stapled peptides
In some embodiments, the provided agent is or comprises a peptide. In some embodiments, the provided agent is a peptide. In some embodiments, the peptide is a stapled peptide. In some embodiments, the provided agent is a stapled peptide. In some embodiments, the peptide is a suture peptide. In some embodiments, the provided agent is a sutures peptide. In some embodiments, the suture peptide comprises two or more staple-like structures, wherein both staple-like structures are bonded to the same peptide backbone atom. A stapled peptide as described herein is typically a peptide in which two or more amino acids of a peptide chain are linked by a linkage of two peptide backbone atoms of amino acid residues, and as understood by those skilled in the art, the linkage is not by a peptide backbone between the linked amino acid residues. In some embodiments, the staple-like structure as described herein is a linker that connects one amino acid residue to another amino acid residue, e.g., by bonding to the peptide backbone atoms of each amino acid residue, and, as understood by those skilled in the art, the connection is through the staple-like structure rather than through the peptide backbone between the connected amino acid residues. In some embodiments, the staple-like structure is bonded to the peptide backbone by replacing one or more hydrogens and/or substituents (e.g., side chains, O, S, etc.) on the peptide backbone atom (e.g., C, N, etc.). In some embodiments, the side chains form part of a staple-like structure. In some embodiments, the staple-like structure is bonded to two carbon backbone atoms (e.g., two alpha carbon atoms). In some embodiments, the staple-like structure comprises C (R') 2 Or N (R'),where R' is R and is bonded to another group attached to the backbone atom (which may be R (e.g., R a3 ) Together with intervening atoms to form a ring as described herein (e.g., when PyrS2 is stapled in a variety of peptides).
In some embodiments, the stapled peptide comprises one or more staple-like structures. In some embodiments, the stapled peptide comprises two or more staple-like structures. In some embodiments, the stapled peptide comprises three or more staple-like structures. In some embodiments, the stapled peptide comprises four or more staple-like structures. In some embodiments, there are three staple-like structures in the stapled peptide. In some embodiments, there are four staple-like structures in the stapled peptide.
As will be appreciated by those of ordinary skill in the art, a variety of peptide stapling techniques are available, including both hydrocarbon stapling techniques and non-hydrocarbon stapling techniques, and may be used in accordance with the present disclosure. Various techniques for stapling and suturing peptides, including various staple-like structures and/or methods for manufacturing are available and may be used in accordance with the present disclosure, such as those described in WO 2019/051327 and WO 2020/04270, the staple-like structures in each of which are incorporated herein by reference.
In some embodiments, the peptide (e.g., stapled peptide) is or comprises a helical structure. In some embodiments, the peptide is a stapled peptide.
In some embodiments, the staple-like structure is a hydrocarbon staple-like structure. In some embodiments, the staple-like structures described herein are non-hydrocarbon staple-like structures. In some embodiments, the non-hydrocarbon staple-like structure comprises one or more chain heteroatoms, wherein the chain of the staple-like structure is the shortest covalent link within the staple-like structure from one end of the staple-like structure to the other end of the staple-like structure. In some embodiments, the non-hydrocarbon staple structure is or comprises at least one sulfur atom derived from an amino acid residue of a polypeptide.In some embodiments, the non-hydrocarbon staple-like structure comprises two sulfur atoms derived from two different amino acid residues of the polypeptide. In some embodiments, the non-hydrocarbon staple-like structure comprises two sulfur atoms derived from two different cysteine residues of the polypeptide. In some embodiments, the staple-like structure is a cysteine staple-like structure. In some embodiments, the staple-like structure is a non-cysteine staple-like structure. In some embodiments, the non-hydrocarbon staple structure is a urethane staple structure and comprises a urethane moiety (e.g., -N (R') -C (O) -O-) in its chain. In some embodiments, the non-hydrocarbon staple-like structure is an amino staple-like structure and comprises amino groups (e.g., -N (R')) in its chain. In some embodiments, the amino group in the amino staple structure, e.g., (-N (R ') -), is not bonded to a carbon atom (e.g., -C (=o), -C (=s), -C (=n-R'), etc.) that otherwise forms a double bond with the heteroatom, such that it is not part of an additional nitrogen-containing group (e.g., an amide, a carbamate, etc.). In some embodiments, the non-hydrocarbon staple structure is an ester staple structure and comprises an ester moiety (-C (O) -O-) in its chain. In some embodiments, the non-hydrocarbon staple structure is an amide staple structure and comprises an amide moiety (-C (O) -N (R')) in its chain. In some embodiments, the non-hydrocarbon staple-like structure is a sulfonamide staple-like structure and comprises sulfonamide moieties (-S (O) in its chain 2 -N (R') (-). In some embodiments, the non-hydrocarbon staple structure is an ether staple structure and comprises an ether moiety (-O-) in its chain. In some embodiments, R' of the carbamate moiety, amino group, amide moiety, sulfonamide moiety, or ether moiety is R, and is attached to an R group of the backbone (e.g., R when it is R a3 ) And intervening atoms therebetween, together to form a ring as described herein. In some embodiments, R' of the carbamate moiety or amino group is R, and is attached to the R group of the backbone (e.g., R when it is R a3 ) And intervening atoms therebetween, together to form a ring as described herein.
In some embodiments, the staple-like structure comprisesOne or more amino groups, such as-N (R ') -, wherein each R' is independently as described herein. In some embodiments, -N (R'), -is bonded to two carbon atoms. In some embodiments, -N (R') -is bonded to two carbon atoms, wherein neither carbon atom is bonded to any heteroatom by a double bond. In some embodiments, -N (R') -is bonded to two sp3 carbon atoms. In some embodiments, the staple-like structure comprises one or more-C (O) -N (R ') -groups, wherein each R' is independently as described herein. In some embodiments, the staple-like structure comprises one or more urethane groups, such as one or more- (O) -C (O) -N (R ') -, wherein each R' is independently as described herein. In some embodiments, R' is-H. In some embodiments, R' is optionally substituted C 1-6 Aliphatic series. In some embodiments, R' is optionally substituted C 1-6 An alkyl group. In some embodiments, R' is C 1-6 Aliphatic series. In some embodiments, R' is C 1-6 An alkyl group. In some embodiments, R' is methyl.
In some embodiments, the stapled peptide comprises one or more staple-like structures. In some embodiments, the stapled peptide comprises one and no more than one staple-like structure. In some embodiments, the stapled peptide comprises two and no more than two staple-like structures. In some embodiments, the two staple-like structures of the stapled peptide are bonded to a common backbone atom. In some embodiments, the two staple-like structures of the stapled peptide are bonded to a common backbone atom that is an alpha carbon atom of an amino acid residue. In some embodiments, the stapled peptide comprises three or more staple-like structures. In some embodiments, the stapled peptide comprises four or more staple-like structures. In some embodiments, the stapled peptide comprises three and no more than three staple-like structures. In some embodiments, the stapled peptide comprises four and no more than four staple-like structures. In some embodiments, each staple-like structure independently has-L as described herein s1 -L s2 -L s3 -a structure. In some embodiments, each staple structure is independently bonded to two amino acid residues. In some embodiments, each staple-like structure is independently bonded to two alpha carbon atoms.
In some embodiments, two, three, four, or all of the staple-like structures in the stapled peptide are located within a region that is several amino acid residues in length. In some embodiments, two staple-like structures are within such an area. In some embodiments, three staple-like structures are within such an area. In some embodiments, four staple-like structures are within such an area. In some embodiments, all of the staple-like structures are within such an area. In some embodiments, the length of the region is 5 to 20, 5 to 15, 5 to 14, 5 to 113, 5 to 12, 5 to 11, 5 to 10, 6 to 20, 6 to 15, 6 to 14, 6 to 113, 6 to 12, 6 to 11, 6 to 10, 7 to 20, 7 to 15, 7 to 14, 7 to 113, 7 to 12, 7 to 11, 7 to 10, 10 to 16, 10 to 15, 10 to 14, 11 to 16, 11 to 15, 11 to 14, 12 to 16, 12 to 15, 12 to 14, 13 to 15, or 13 to 14 amino acid residues. In some embodiments, the region is 5 amino acid residues in length. In some embodiments, the region is 6 amino acid residues in length. In some embodiments, the region is 7 amino acid residues in length. In some embodiments, the region is 8 amino acid residues in length. In some embodiments, the region is 9 amino acid residues in length. In some embodiments, the region is 10 amino acid residues in length. In some embodiments, the region is 11 amino acid residues in length. In some embodiments, the region is 12 amino acid residues in length. In some embodiments, the region is 13 amino acid residues in length. In some embodiments, the region is 14 amino acid residues in length. In some embodiments, the region is 15 amino acid residues in length. In some embodiments, the region is 16 amino acid residues in length. In some embodiments, the region is 17 amino acid residues in length. In some embodiments, the region is 18 amino acid residues in length. In some embodiments, the region is 19 amino acid residues in length. In some embodiments, the region is 20 amino acid residues in length. For example, in various embodiments, the stapled peptide comprises three staple-like structures within a14 amino acid region (e.g., a staple-like structure bonded to aa1 and aa4, a staple-like structure bonded to aa4 and aa11, and a staple-like structure bonded to aa10 and aa 14).
In some embodiments, the peptide of the present disclosure (e.g., a stapled peptide) is or comprises a helical structure. As will be appreciated by those skilled in the art, the spiral may have a variety of lengths. In some embodiments, the helices are 5 to 30 amino acid residues in length. In some embodiments, the length of the helix is 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 or more amino acid residues. In some embodiments, the length of the helix is 6 amino acid residues. In some embodiments, the length of the helix is 8 amino acid residues. In some embodiments, the length of the helix is 10 amino acid residues. In some embodiments, the length of the helix is 12 amino acid residues. In some embodiments, the length of the helix is 14 amino acid residues. In some embodiments, the length of the helix is 16 amino acid residues. In some embodiments, the helix is 17 amino acid residues in length. In some embodiments, the length of the helix is 18 amino acid residues. In some embodiments, the length of the helix is 19 amino acid residues. In some embodiments, the helix is 20 amino acid residues in length.
The amino acids stapled together may have a variety of numbers of amino acid residues between them, such as 1 to 20, 1 to 15, 1 to 10, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, etc. In some embodiments, the staple-like structure is (i, i+4), which means that there are three amino acid residues (at positions i+1, i+2, i+3, respectively) between the two amino acids (at positions i and i+4, respectively) to which the staple-like structure is bonded. In some embodiments, the staple-like structure is (i, i+2). In some embodiments, the staple-like structure is (i, i+3). In some embodiments, the staple-like structure is (i, i+5). In some embodiments, the staple-like structure is (i, i+6). In some embodiments, the staple-like structure is (i, i+7). In some embodiments, the staple-like structure is (i, i+8). In some embodiments, the stapled peptide comprises two staple-like structures, one is (i, i+2) and the other is (i, i+7). In some embodiments, the stapled peptide comprises two staple-like structures, one is (i, i+3) and the other is (i, i+7). In some embodiments, the stapled peptide comprises two staple-like structures, one is (i, i+3) and the other is (i, i+4). In some embodiments, the stapled peptide comprises two staple-like structures, one being (i, i+4) and the other being (i, i+7). In some embodiments, the stapled peptide comprises two staple-like structures, one being (i, i+3) and the other being (i, i+3). In some embodiments, the stapled peptide comprises two staple-like structures, one being (i, i+4) and the other being (i, i+4). In some embodiments, the stapled peptide comprises two staple-like structures, one being (i, i+7) and the other being (i, i+7). In some embodiments, two staple-like structures are bonded to a common backbone atom, such as an alpha carbon atom of an amino acid residue. In some embodiments, the stapled peptide further comprises a third staple-like structure. In some embodiments, the third staple-like structure is (i, i+3). In some embodiments, the third staple-like structure is (i, i+4). In some embodiments, the third staple-like structure is (i, i+7). In some embodiments, the stapled peptide further comprises a fourth staple-like structure. In some embodiments, the fourth staple-like structure is (i, i+3). In some embodiments, the fourth staple-like structure is (i, i+4). In some embodiments, the fourth staple-like structure is (i, i+7).
In some embodiments, the stapled peptide comprises a staple-like structure, which is L s Wherein L is s is-L s1 -L s2 -L s3 -，L s1 、L s2 And L s3 Each independently is L, wherein each L is independentlyAs described in this disclosure. In some embodiments, the staple-like structure provided is L s 。
In some embodiments, L s1 Comprising at least one-N (R ') -wherein R' is as described in the present disclosure. In some embodiments, -N (R') -is bonded to two carbon atoms, wherein neither carbon atom forms a double bond with a heteroatom. In some embodiments, -N (R') -is not bonded to-C (O) -. In some embodiments, -N (R') -is not bonded to-C (S) -. In some embodiments, -N (R ') -is not bonded to-C (=nr') -. In some embodiments, L s1 is-L ' -N (R ') -, wherein L ' is an optionally substituted divalent C 1 -C 19 Aliphatic series. In some embodiments, L s1 is-L' -N (CH) 3 ) -, wherein L' is an optionally substituted divalent C 1 -C 19 Aliphatic series.
In some embodiments, R' is optionally substituted C 1-6 An alkyl group. In some embodiments, R' is C 1-6 An alkyl group. In some embodiments, R' is methyl. In some embodiments, with L s1 The peptide skeleton atoms being bound to R 1 Bonded, and R' and R 1 Both are R and taken together with their intervening atoms form an optionally substituted ring as described in this disclosure. In some embodiments, the formed ring has no additional ring heteroatoms other than the nitrogen atom to which R' is bonded. In some embodiments, the ring formed is 3-membered. In some embodiments, the ring formed is 4-membered. In some embodiments, the ring formed is 5-membered. In some embodiments, the ring formed is 6 membered.
In some embodiments, L' is an optionally substituted divalent C 1 -C 20 Aliphatic series. In some embodiments, L' is an optionally substituted divalent C 1 -C 19 Aliphatic series. In some embodiments, L' is an optionally substituted divalent C 1 -C 15 Aliphatic series. In some embodiments, L' is an optionally substituted divalent C 1 -C 10 Aliphatic series. In some embodimentsIn which L' is an optionally substituted divalent C 1 -C 9 Aliphatic series. In some embodiments, L' is an optionally substituted divalent C 1 -C 8 Aliphatic series. In some embodiments, L' is an optionally substituted divalent C 1 -C 7 Aliphatic series. In some embodiments, L' is an optionally substituted divalent C 1 -C 6 Aliphatic series. In some embodiments, L' is an optionally substituted divalent C 1 -C 5 Aliphatic series. In some embodiments, L' is an optionally substituted divalent C 1 -C 4 Aliphatic series. In some embodiments, L' is optionally substituted alkylene. In some embodiments, L' is optionally substituted alkenylene. In some embodiments, L' is unsubstituted alkylene. In some embodiments, L' is-CH 2 -. In some embodiments, L' is- (CH) 2 ) 2 -. In some embodiments, L' is- (CH) 2 ) 3 -. In some embodiments, L' is- (CH) 2 ) 4 -. In some embodiments, L' is- (CH) 2 ) 5 -. In some embodiments, L' is- (CH) 2 ) 6 -. In some embodiments, L' is- (CH) 2 ) 7 -. In some embodiments, L' is- (CH) 2 ) 8 -. In some embodiments, L' is bonded to a peptide backbone atom. In some embodiments, L' is optionally substituted alkenylene. In some embodiments, L' is unsubstituted alkenylene. In some embodiments, L' is-CH 2 -CH＝CH-CH 2 -。
In some embodiments, L' is optionally substituted phenylene.
In some embodiments, L s1 Comprising at least one-N (R ') C (O) -, wherein R' is as described in the present disclosure. In some embodiments, L s1 is-L '-N (R') C (O) -, wherein L 'and R' are each independently as described in the disclosure. In some embodiments, L s1 is-L' -N (CH) 3 ) C (O) -, wherein L' is independently as described in the present disclosure.
In some embodiments, L s1 Comprising at least one-C (O) O-. In some embodiments, L s1 Comprising at least one-C (O) O-. In some embodiments, L s1 is-L ' -C (O) O-or-L ' -OC (O) -, wherein each L ' is independently as described in the present disclosure. In some embodiments, L s1 is-L '-C (O) O-, wherein each L' is independently as described in the present disclosure. In some embodiments, L s1 is-L '-OC (O) -, wherein each L' is independently as described in the present disclosure.
In some embodiments, L s1 Comprising at least one of-S (O) 2 -N (R ') -, wherein R' is as described in the present disclosure. In some embodiments, L s1 Comprising at least one of-S (O) 2 -N (R ') -, wherein R' is as described in the present disclosure. In some embodiments, L s1 is-L '-N (R') -S (O) 2 -or-L' -S (O) 2 -N (R ') -, wherein L ' and R ' are each independently as described in the present disclosure. In some embodiments, L s1 is-L '-N (R') -S (O) 2 -wherein L 'and R' are each independently as described in the present disclosure. In some embodiments, L s1 is-L' -S (O) 2 -N (R ') -, wherein L ' and R ' are each independently as described in the present disclosure. In some embodiments, L s1 is-L' -N (CH) 3 )-S(O) 2 -or-L' -S (O) 2 -N(CH 3 ) -, wherein each L' is independently as described in the present disclosure. In some embodiments, L s1 is-L' -N (CH) 3 )-S(O) 2 -, wherein L' is as described in the present disclosure. In some embodiments, L s1 is-L' -S (O) 2 -N(CH 3 ) -, wherein L' is as described in the present disclosure.
In some embodiments, L s1 Comprising at least one-O-. In some embodiments, L s1 is-L '-O-, wherein L' is independently as described in the present disclosure.
In some embodiments, L s1 Is a covalent bond.
In some casesIn embodiments, L s1 Is L ', wherein L' is as described in the present disclosure.
In some embodiments, L s2 Is L, wherein L is as described in the present disclosure. In some embodiments, L s2 Is L ', wherein L' is as described in the present disclosure. In some embodiments, L s2 comprises-CH 2 -CH＝CH-CH 2 -. In some embodiments, L s2 is-CH 2 -CH＝CH-CH 2 -. In some embodiments, L s2 Comprises- (CH) 2 ) 4 -. In some embodiments, L s2 Is- (CH) 2 ) 4 -。
In some embodiments, L s3 Comprising at least one-N (R ') -wherein R' is as described in the present disclosure. In some embodiments, -N (R') -is bonded to two carbon atoms, wherein neither carbon atom forms a double bond with a heteroatom. In some embodiments, -N (R') -is not bonded to-C (O) -. In some embodiments, -N (R') -is not bonded to-C (S) -. In some embodiments, -N (R ') -is not bonded to-C (=nr') -. In some embodiments, L s3 is-L ' -N (R ') -, wherein L ' is an optionally substituted divalent C 1 -C 19 Aliphatic series. In some embodiments, L s3 is-L' -N (CH) 3 ) -, wherein L' is an optionally substituted divalent C 1 -C 19 Aliphatic series.
In some embodiments, L s3 Comprising at least one-N (R ') C (O) -, wherein R' is as described in the present disclosure. In some embodiments, L s3 is-L '-N (R') C (O) -, wherein L 'and R' are each independently as described in the disclosure. In some embodiments, L s3 is-L' -N (CH) 3 ) C (O) -, wherein L' is independently as described in the present disclosure.
In some embodiments, L s3 Comprising at least one-C (O) O-. In some embodiments, L s3 Comprising at least one-C (O) O-. In some embodiments, L s3 is-L ' -C (O) O-or-L ' -OC (O) -, wherein each L ' is independently as in the present disclosureSaid method. In some embodiments, L s3 is-L '-C (O) O-, wherein each L' is independently as described in the present disclosure. In some embodiments, L s3 is-L '-OC (O) -, wherein each L' is independently as described in the present disclosure.
In some embodiments, L s3 Comprising at least one of-S (O) 2 -N (R ') -, wherein R' is as described in the present disclosure. In some embodiments, L s3 Comprising at least one of-S (O) 2 -N (R ') -, wherein R' is as described in the present disclosure. In some embodiments, L s3 is-L '-N (R') -S (O) 2 -or-L' -S (O) 2 -N (R ') -, wherein L ' and R ' are each independently as described in the present disclosure. In some embodiments, L s3 is-L '-N (R') -S (O) 2 -wherein L 'and R' are each independently as described in the present disclosure. In some embodiments, L s3 is-L' -S (O) 2 -N (R ') -, wherein L ' and R ' are each independently as described in the present disclosure. In some embodiments, L s3 is-L' -N (CH) 3 )-S(O) 2 -or-L' -S (O) 2 -N(CH 3 ) -, wherein each L' is independently as described in the present disclosure. In some embodiments, L s3 is-L' -N (CH) 3 )-S(O) 2 -, wherein L' is as described in the present disclosure. In some embodiments, L s3 is-L' -S (O) 2 -N(CH 3 ) -, wherein L' is as described in the present disclosure.
In some embodiments, L s3 Comprising at least one-O-. In some embodiments, L s3 is-L '-O-, wherein L' is independently as described in the present disclosure.
In some embodiments, L s3 Is L ', wherein L' is as described in the present disclosure. In some embodiments, L s3 Is an optionally substituted alkylene group. In some embodiments, L s3 Is an unsubstituted alkylene group.
In some embodiments, L s Comprising at least one-N (R ') -wherein R' is as described in the present disclosure.In some embodiments, -N (R') -is bonded to two carbon atoms, wherein neither carbon atom forms a double bond with a heteroatom. In some embodiments, -N (R') -is not bonded to-C (O) -. In some embodiments, -N (R') -is not bonded to-C (S) -. In some embodiments, -N (R ') -is not bonded to-C (=nr') -. In some embodiments, L s Comprising at least one-N (R ') C (O) -, wherein R' is as described in the present disclosure.
In some embodiments, L s 、L s1 、L s2 And L s3 Each independently and optionally containing an R 'group, e.g., -C (R') 2 -R ' groups in-N (R ') -and the like, and the R ' groups are bonded to groups (e.g., groups may be R) (e.g., L a1 Or L a2 R of (2) a1 、R a2 、R a3 R 'groups (e.g., -C (R') 2 -R 'groups in-N (R') -etc.), together to form a double bond or an optionally substituted ring, as may be formed by two R groups. In some embodiments, the ring formed is an optionally substituted 3-to 10-membered ring. In some embodiments, the ring formed is an optionally substituted 3-membered ring. In some embodiments, the ring formed is an optionally substituted 4-membered ring. In some embodiments, the ring formed is an optionally substituted 5-membered ring. In some embodiments, the ring formed is an optionally substituted 6 membered ring. In some embodiments, the ring formed is a single ring. In some embodiments, the formed ring is saturated. In some embodiments, the rings formed are partially unsaturated. In some embodiments, the ring formed is aromatic. In some embodiments, the formed ring comprises one or more ring heteroatoms (e.g., nitrogen). In some embodiments, the staple-like structure or L s 、L s1 、L s2 And/or L s3 comprises-N (R ') -, and R' together with the group attached to the backbone atom form an optionally substituted ring as described herein. In some embodiments, the staple-like structure or L s 、L s1 、L s2 And/or L s3 comprising-C (R') 2 -, R' is attached toThe groups attached to the backbone atoms are taken together to form an optionally substituted ring as described herein.
In some embodiments, the staple-like structure or L s 、L s1 、L s2 And/or L s3 A portion comprising one or more amino acid side chains (e.g., except for its terminal = CH 2 External side chains).
As will be apparent to those skilled in the art upon reading this disclosure, the letter "L" is used to refer to a linker moiety as described herein; thus, in some embodiments, unless otherwise indicated, each L Superscript (e.g., L a 、L s1 、L s2 、L s3 、L s Etc.) is understood to be L.
In some embodiments, L comprises at least one —n (R ') -, wherein R' is as described in the present disclosure. In some embodiments, -N (R') -is bonded to two carbon atoms, wherein neither carbon atom forms a double bond with a heteroatom. In some embodiments, -N (R') -is not bonded to-C (O) -. In some embodiments, -N (R') -is not bonded to-C (S) -. In some embodiments, -N (R ') -is not bonded to-C (=nr') -. In some embodiments, L is-L ' -N (R ') -, wherein L ' is an optionally substituted divalent C 1 -C 19 Aliphatic series. In some embodiments, L is-L' -N (CH 3 ) -, wherein L' is an optionally substituted divalent C 1 -C 19 Aliphatic series.
In some embodiments, L comprises at least one-N (R ') C (O) -, wherein R' is as described in the present disclosure. In some embodiments, L is-L '-N (R') C (O) -, wherein L 'and R' are each independently as described in the disclosure. In some embodiments, L is-L' -N (CH 3 ) C (O) -, wherein L' is independently as described in the present disclosure.
In some embodiments, L comprises at least one-C (O) O-. In some embodiments, L comprises at least one-C (O) O-. In some embodiments, L is-L ' -C (O) O-or-L ' -OC (O) -, wherein each L ' is independently as described in the present disclosure. In some embodiments, L is-L '-C (O) O-, wherein each L' is independently as described in the present disclosure. In some embodiments, L is-L '-OC (O) -, wherein each L' is independently as described in the present disclosure.
In some embodiments, L comprises at least one of-S (O) 2 -N (R ') -, wherein R' is as described in the present disclosure. In some embodiments, L comprises at least one of-S (O) 2 -N (R ') -, wherein R' is as described in the present disclosure. In some embodiments, L is-L '-N (R') -S (O) 2 -or-L' -S (O) 2 -N (R ') -, wherein L ' and R ' are each independently as described in the present disclosure. In some embodiments, L is-L '-N (R') -S (O) 2 -wherein L 'and R' are each independently as described in the present disclosure. In some embodiments, L is-L' -S (O) 2 -N (R ') -, wherein L ' and R ' are each independently as described in the present disclosure. In some embodiments, L is-L' -N (CH 3 )-S(O) 2 -or-L' -S (O) 2 -N(CH 3 ) -, wherein each L' is independently as described in the present disclosure. In some embodiments, L is-L' -N (CH 3 )-S(O) 2 -, wherein L' is as described in the present disclosure. In some embodiments, L is-L' -S (O) 2 -N(CH 3 ) -, wherein L' is as described in the present disclosure.
In some embodiments, L comprises at least one-O-. In some embodiments, L is-L '-O-, wherein L' is independently as described in the present disclosure.
In some embodiments, L is L ', wherein L' is as described in the present disclosure. In some embodiments, L is optionally substituted alkylene. In some embodiments, L is unsubstituted alkylene.
In some embodiments, L is an optionally substituted divalent C 1 -C 25 Aliphatic series. In some embodiments, L is an optionally substituted divalent C 1 -C 20 Aliphatic series. In some embodiments, L is an optionally substituted divalent C 1 -C 15 Aliphatic series. In some embodiments, L is optionally substitutedDivalent C 1 -C 10 Aliphatic series. In some embodiments, L is an optionally substituted divalent C 1 -C 9 Aliphatic series. In some embodiments, L is an optionally substituted divalent C 1 -C 8 Aliphatic series. In some embodiments, L is an optionally substituted divalent C 1 -C 7 Aliphatic series. In some embodiments, L is an optionally substituted divalent C 1 -C 6 Aliphatic series. In some embodiments, L is an optionally substituted divalent C 1 -C 5 Aliphatic series. In some embodiments, L is an optionally substituted divalent C 1 -C 4 Aliphatic series. In some embodiments, L is optionally substituted alkylene. In some embodiments, L is optionally substituted alkenylene. In some embodiments, L is unsubstituted alkylene. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments, L is- (CH) 2 ) 5 -. In some embodiments, L is- (CH) 2 ) 6 -. In some embodiments, L is- (CH) 2 ) 7 -. In some embodiments, L is- (CH) 2 ) 8 -. In some embodiments, L is bonded to a peptide backbone atom. In some embodiments, L is optionally substituted alkenylene. In some embodiments, L is unsubstituted alkenylene. In some embodiments, L is-CH 2 -CH＝CH-CH 2 -。
In some embodiments, one end of the staple-like structure is attached to atom A of the peptide backbone n1 Connection, wherein A n1 Optionally by R 1 Substituted and is the amino acid position N from the N-terminus of the peptide 1 An atom of an amino acid residue at the site, and the other end is attached to an atom A of the peptide backbone n2 Connection, wherein A n2 Optionally by R 2 (in some embodiments, R) 1 And/or R 2 R, which may be hydrogen) and is the amino acid position N from the N-terminus of the peptide 2 An atom of an amino acid residue at which n 1 And n 2 Each independently is an integer, and n 2 ＝n 1 +m, where m is 3 to 12.
In some embodiments, m is 3. In some embodiments, m is 4. In some embodiments, m is 5. In some embodiments, m is 6. In some embodiments, m is 7. In some embodiments, m is 8. In some embodiments, m is 9. In some embodiments, m is 10. In some embodiments, m is 11. In some embodiments, the staple-like structure is referred to as an (i, i+m) staple-like structure.
In some embodiments, a n1 Is a carbon atom. In some embodiments, a n1 Is achiral. In some embodiments, a n1 Is chiral. In some embodiments, a n1 Is R. In some embodiments, a n1 Is S.
In some embodiments, a n2 Is a carbon atom. In some embodiments, a n2 Is achiral. In some embodiments, a n2 Is chiral. In some embodiments, a n2 Is R. In some embodiments, a n2 Is S.
In some embodiments, a n1 Is achiral and A n2 Is achiral. In some embodiments, a n1 Is achiral and A n2 Is R. In some embodiments, a n1 Is achiral and A n2 Is S. In some embodiments, a n1 Is R and A n2 Is achiral. In some embodiments, a n1 Is R and A n2 Is R. In some embodiments, a n1 Is R and A n2 Is S. In some embodiments, a n1 Is S and A n2 Is achiral. In some embodiments, a n1 Is S and A n2 Is R. In some embodiments, a n1 Is S and A n2 Is S.
In some embodiments, the stereochemistry and/or combinations thereof provided at the staple-structure-backbone junction optionally in combination with one or more structural elements of the provided peptides (e.g., staple-structure chemicals (hydrocarbons, non-hydrocarbons), staple-structure lengths, etc.) can provide a variety of benefits, such as improved production yield, purity and/or selectivity, improved properties (e.g., improved solubility, improved stability, reduced toxicity, improved selectivity, etc.), improved activity, etc. In some embodiments, the provided stereochemistry and/or stereochemical combinations are different from those commonly used, e.g., those in US 9617309, US2015-0225471, US2016-0024153, US2016-0215036, US 2016-024494, WO 2017/062518, and provide one or more of the benefits described in the present disclosure.
In some embodiments, the staple-like structure may have a variety of lengths, in some embodiments as represented by the number of chain atoms of the staple-like structure. In some embodiments, the chain of the staple-like structure is the shortest covalent link in the staple-like structure from a first end of the staple-like structure (the point of attachment to the peptide backbone) to a second end of the staple-like structure, wherein the first end and the second end are attached to two different peptide backbone atoms. In some embodiments, the staple-like structure comprises 5 to 30 chain atoms, for example 5 to 20, 5 to 15, 5, 6, 7, 8, 9 or 10 to 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 chain atoms. In some embodiments, the staple-like structure comprises 5 chain atoms. In some embodiments, the staple-like structure comprises 6 chain atoms. In some embodiments, the staple-like structure comprises 7 chain atoms. In some embodiments, the staple-like structure comprises 8 chain atoms. In some embodiments, the staple-like structure comprises 9 chain atoms. In some embodiments, the staple-like structure comprises 10 chain atoms. In some embodiments, the staple-like structure comprises 11 chain atoms. In some embodiments, the staple-like structure comprises 12 chain atoms. In some embodiments, the staple-like structure comprises 13 chain atoms. In some embodiments, the staple-like structure comprises 14 chain atoms. In some embodiments, the staple-like structure comprises 15 chain atoms. In some embodiments, the staple-like structure comprises 16 chain atoms. In some embodiments, the staple-like structure comprises 17 chain atoms. In some embodiments, the staple-like structure comprises 18 chain atoms. In some embodiments, the staple-like structure comprises 19 chain atoms. In some embodiments, the staple-like structure comprises 20 chain atoms. In some embodiments, the staple-like structure is 5 chain atoms in length. In some embodiments, the staple-like structure is 6 chain atoms in length. In some embodiments, the staple-like structure is 7 chain atoms in length. In some embodiments, the staple-like structure is 8 chain atoms in length. In some embodiments, the staple-like structure is 9 chain atoms in length. In some embodiments, the staple-like structure is 10 chain atoms in length. In some embodiments, the staple-like structure is 11 chain atoms in length. In some embodiments, the staple-like structure is 12 chain atoms in length. In some embodiments, the staple-like structure is 13 chain atoms in length. In some embodiments, the staple-like structure is 14 chain atoms in length. In some embodiments, the staple-like structure is 15 chain atoms in length. In some embodiments, the staple-like structure is 16 chain atoms in length. In some embodiments, the staple-like structure is 17 chain atoms in length. In some embodiments, the staple-like structure is 18 chain atoms in length. In some embodiments, the staple-like structure is 19 chain atoms in length. In some embodiments, the staple-like structure is 20 chain atoms in length. In some embodiments, the staple-like structure is 8 to 15 chain atoms in length. In some embodiments, the staple-like structure has 8 to 12 chain atoms. In some embodiments, the staple-like structure has 9 to 12 chain atoms. In some embodiments, the staple-like structure has 9 to 10 chain atoms. In some embodiments, the staple-like structure has 8 to 10 chain atoms. In some embodiments, the length of the staple-like structure may be adjusted according to the distance of the amino acid residues to which it is attached, e.g., longer staple-like structures may be used for (i, i+7) compared to (i, i+4) or (i, i+3) staple-like structures. In some embodiments, the (i, i+2) staple-like structure has about 5 to 10, 5 to 8, e.g., about 5, 6, 7, 8, 9, or 10 chain atoms. In some embodiments, the (i, i+2) staple-like structure has 5 chain atoms. In some embodiments, the (i, i+2) staple-like structure has 6 chain atoms. In some embodiments, the (i, i+2) staple-like structure has 7 chain atoms. In some embodiments, the (i, i+2) staple-like structure has 8 chain atoms. In some embodiments, the (i, i+2) staple-like structure has 9 chain atoms. In some embodiments, the (i, i+2) staple-like structure has 10 chain atoms. In some embodiments, the (i, i+3) staple-like structure has about 5 to 10, 5 to 8, e.g., about 5, 6, 7, 8, 9, or 10 chain atoms. In some embodiments, the (i, i+3) staple-like structure has 5 chain atoms. In some embodiments, the (i, i+3) staple-like structure has 6 chain atoms. In some embodiments, the (i, i+3) staple-like structure has 7 chain atoms. In some embodiments, the (i, i+3) staple-like structure has 8 chain atoms. In some embodiments, the (i, i+3) staple-like structure has 9 chain atoms. In some embodiments, the (i, i+3) staple-like structure has 10 chain atoms. In some embodiments, the (i, i+4) staple-like structure has about 5 to 12, 5 to 10, 7 to 12, 5 to 8, e.g., about 5, 6, 7, 8, 9, 10, 11, or 12 chain atoms. In some embodiments, the (i, i+4) staple-like structure has 5 chain atoms. In some embodiments, the (i, i+4) staple-like structure has 6 chain atoms. In some embodiments, the (i, i+4) staple-like structure has 7 chain atoms. In some embodiments, the (i, i+4) staple-like structure has 8 chain atoms. In some embodiments, the (i, i+4) staple-like structure has 9 chain atoms. In some embodiments, the (i, i+4) staple-like structure has 10 chain atoms. In some embodiments, the (i, i+4) staple-like structure has 11 chain atoms. In some embodiments, the (i, i+4) staple-like structure has 12 chain atoms. In some embodiments, the (i, i+7) staple-like structure has about 8 to 25, 10 to 16, 12 to 15, e.g., about 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 chain atoms. In some embodiments, the (i, i+7) staple-like structure has 8 chain atoms. In some embodiments, the (i, i+7) staple-like structure has 9 chain atoms. In some embodiments, the (i, i+7) staple-like structure has 10 chain atoms. In some embodiments, the (i, i+7) staple-like structure has 11 chain atoms. In some embodiments, the (i, i+7) staple-like structure has 12 chain atoms. In some embodiments, the (i, i+7) staple-like structure has 13 chain atoms. In some embodiments, the (i, i+7) staple-like structure has 14 chain atoms. In some embodiments, the (i, i+7) staple-like structure has 15 chain atoms. In some embodiments, the (i, i+7) staple-like structure has 16 chain atoms. In some embodiments, the (i, i+7) staple-like structure has 17 chain atoms. In some embodiments, the (i, i+7) staple-like structure has 18 chain atoms. In some embodiments, the (i, i+7) staple-like structure has 19 chain atoms. In some embodiments, the (i, i+7) staple-like structure has 20 chain atoms. In some embodiments, the (i, i+7) staple-like structure has 21 chain atoms. In some embodiments, the (i, i+7) staple-like structure has 22 chain atoms. In some embodiments, the stapled peptide comprises three or more staple-like structures, each of which is independently such an (i, i+2), (i, i+3), (i, i+4), or (i, i+7) staple-like structure. In some embodiments, the stapled peptide comprises such an (i, i+2) staple-like structure, such an (i, i+4) staple-like structure, and such an (i, i+7) staple-like structure. In some embodiments, the stapled peptide comprises such an (i, i+3) staple-like structure, such an (i, i+4) staple-like structure, and such an (i, i+7) staple-like structure. In some embodiments, the stapled peptide comprises such an (i, i+3) staple-like structure, such an (i, i+7) staple-like structure, and such an (i, i+7) staple-like structure.
The staple-like structure length may be described in further detail. For example, in some embodiments, the staple-like structure length is described as the total number of chain atoms and non-chain ring atoms, where a non-chain ring atom is an atom of the staple-like structure that forms a ring with one or more chain atoms, but is not a chain atom, as it is not within the shortest covalent connection from the first end of the staple-like structure to the second end of the staple-like structure. In some embodiments, the staple-like structure formed using monomer a (which comprises an azetidine moiety), monomer B (which comprises a pyrrolidine moiety), and/or monomer C (which comprises a pyrrolidine moiety), and the like, may comprise one or two non-chain ring atoms.
In some embodiments, the staple-like structure does not have heteroatoms in its chain. In some embodiments, the staple-like structure comprises at least one heteroatom in its chain. In some embodiments, the staple-like structure comprises at least one nitrogen atom in its chain.
In some embodiments, the staple-like structure is L s Wherein L is s Is an optionally substituted divalent C 8-14 An aliphatic group, wherein one or more methylene units of the aliphatic group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, the staple-like structure is L s Wherein L is s Is an optionally substituted divalent C 9-13 An aliphatic group, wherein one or more methylene units of the aliphatic group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, the staple-like structure is L s Wherein L is s Is an optionally substituted divalent C 10-15 Aliphatic seriesA group wherein one or more methylene units of the aliphatic group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, the staple-like structure is L s Wherein L is s Is an optionally substituted divalent C 11-14 An aliphatic group, wherein one or more methylene units of the aliphatic group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, the staple-like structure is an (i, i+2) staple-like structure, wherein two amino acid residues directly linked to the staple-like structure are not included, with one amino acid residue between the two amino acid residues directly linked to the staple-like structure. In some embodiments, the staple-like structure is an (i, i+3) staple-like structure, wherein two amino acid residues directly linked to the staple-like structure are not included, with two amino acid residues in between the two amino acid residues directly linked to the staple-like structure. In some embodiments, the staple-like structure is an (i, i+4) staple-like structure, wherein two amino acid residues directly linked to the staple-like structure are not included, and there are three amino acid residues between the two amino acid residues directly linked to the staple-like structure. In some embodiments, the staple-like structure is an (i, i+7) staple-like structure, wherein two amino acid residues directly linked to the staple-like structure are not included, and there are six amino acid residues between the two amino acid residues directly linked to the staple-like structure.
In some embodiments, for L s ，L s1 ，L s2 And L s3 In each of the above-mentioned steps, any substitution of methylene units, if any, is by-N (R ') -; -C (O) -N (R '), -N (R ') C (O) O-, -C (O) O-, and,-S(O) 2 N (R') -or-O-substitution. In some embodiments, for L s 、L s1 、L s2 And L s3 In each of the above-mentioned steps, any substitution of methylene units, if any, is by-N (R ') -; -N (R ') -C (O) -or-N (R ') C (O) O-substitution. In some embodiments, for L s 、L s1 、L s2 And L s3 Any substitution of methylene units, if any, is substituted by-N (R ') -or-N (R') C (O) O-. In some embodiments, for L s 、L s1 、L s2 And L s3 Any substitution, if any, of the methylene units is substituted by-N (R') -. In some embodiments, for L s 、L s1 、L s2 And L s3 Any substitution of methylene units, if any, is replaced by-N (R') C (O) O-.
In some embodiments, the staple-like structure comprises a double bond. In some embodiments, the staple-like structure comprises a double bond that can be formed by olefin metathesis of two olefins. In some embodiments, the staple-like structure is formed by a metathesis reaction, e.g., involving one or more double bonds in an amino acid residue as described herein. In some embodiments, the first amino acid residue comprising an alkene (e.g., AA 1-ch=ch 2 ) And a second amino acid residue comprising an alkene (e.g., AA 2-ch=ch 2 ) Are stapled (e.g., forming AA 1-ch=ch-AA 2, where AA1 and AA2 are typically connected by one or more amino acid residues). In some embodiments, for example, the olefin in the staple-like structure is converted to-CHR '-, where each R' is independently as described herein. In some embodiments, R' is R, as described herein. In some embodiments, R' is-H. In some embodiments, each R' is-H. In some embodiments, R' is-OR, wherein R is as described herein. In some embodiments, R' is-OH. In some embodiments, R' is-N (R) 2 Wherein each R is independently as described herein. In some embodiments, R' is-SR, wherein R is as described herein. In some casesIn embodiments, R' is R, where R is an optionally substituted aliphatic, e.g., C 1-10 Aliphatic series. In some embodiments, R' is R, where R is an optionally substituted aliphatic, e.g., C 1-10 Alkenyl groups. In some embodiments, R' is R, where R is an optionally substituted aliphatic, e.g., C 1-10 Alkynyl groups. In some embodiments, -CHR '-CHR' -is-CH 2 -CH 2 -. In some embodiments, each of the two olefins is independent of the side chain of the amino acid residue. In some embodiments, each olefin is independently a terminal olefin. In some embodiments, each olefin is independently a monosubstituted olefin.
In some embodiments, the amino acid of formula a-I or salt thereof is a compound having the structure of formula a-II:
NH(R a1 )-L a1 -C(-L a -CH＝CH 2 )(R a3 )-L a2 -COOH，
A-II
wherein each variable is independently as described in the present disclosure. In some embodiments, amino acids suitable for stapling have the structure of formula a-II or salts thereof, wherein each variable is independently as described in the present disclosure.
In some embodiments, the amino acid of formula a-II or salt thereof is a compound having the structure of formula a-II-b or salt thereof:
NH(R a1 )-C(-L a -CH＝CH 2 )(R a3 )-COOH，
A-II-b
wherein each variable is independently as described in the present disclosure. In some embodiments, the amino acid suitable for stapling has the structure of formula a-II-b, or a salt thereof, wherein each variable is independently as described in the present disclosure.
In some embodiments, the amino acids of formula a-I or salts thereof are compounds having the structure of formula a-III:
N(-L a -CH＝CH 2 )(R a1 )-L a1- C(-L a -CH＝CH 2 )(R a3 )-L a2 -COOH，
A-III
wherein each variable is independently as described in the present disclosure. In some embodiments, amino acids suitable for stapling have the structure of formula a-II or salts thereof, wherein each variable is independently as described in the present disclosure.
In some embodiments, the amino acids of formula a-I or salts thereof have the structure of formula a-IV or salts thereof:
NH(R a1 )-L a1 -C(-L a -COOH)(R a3 )-L a2 -COOH,
A-IV
wherein each variable is independently as described in the present disclosure. In some embodiments, amino acids suitable for stapling have the structure of formulas a-IV or salts thereof, wherein each variable is independently as described in the present disclosure.
In some embodiments, the amino acid has the structure of formula a-V or a salt thereof:
NH(R a1 )-L a1 -C(-L a -R SP1 )(R a3 )-I a2 -COOH，
A-V
wherein each variable is independently as described in the present disclosure. In some embodiments, the amino acid suitable for stapling has the formula a-V structure or a salt thereof, wherein each variable is independently as described in the present disclosure.
In some embodiments, the amino acid for stapling has the structure of formulas a-VI or a salt thereof:
NH(R a1 )-L a1 -C(-L a -R SP1 )(-L a -R SP2 )-L a2 -COOH，
A-VI
wherein each variable is independently as described in the present disclosure. In some embodiments, amino acids suitable for stapling have the structure of formulas a-VI or salts thereof, wherein each variable is independently as described in the present disclosure.
R as used herein SP1 And R is SP2 Each independently comprising a reactive group. In some embodiments, R SP1 And R is SP2 Each independently is a reactive group. In some embodiments, the reactive group is an optionally substituted-ch=ch 2 . In some embodiments, the reactive group is-ch=ch 2 . In some embodiments, the reactive group is an amino group, e.g., -NHR, wherein R is as described herein. In some embodiments, the reactive group is an acidic group. In some embodiments, the reactive group is-COOH or an activated form thereof. In some embodiments, the reactive groups are used in cycloaddition reactions (e.g., [3+2 ]]、[4+2]Etc.), e.g., alkene, alkyne, diene, 1, 3-dipole (e.g., -N) 3 ) Etc. In some embodiments, the reactive group is an optionally substituted-c≡ch. In some embodiments, the reactive group is-c≡ch. In some embodiments, the reactive group is-N 3 。
In some embodiments, R of the first amino acid residue SP1 Or R is SP2 And R of a second amino acid residue SP1 And R is SP2 Can react with each other such that two amino acid residues are linked to the staple-like structure. In some embodiments, the reaction is two olefins, e.g., two-ch=ch 2 Olefin metathesis therebetween. In some embodiments, the reaction is amidation and one reactive group is amino, e.g., -NHR, where R is as described herein (e.g., in some embodiments, R is-H; in some embodiments, R is optionally substituted C) 1-6 Aliphatic) and the other is an acidic group (e.g., -COOH) or an activated form thereof. In some embodiments, the reaction is a cycloaddition reaction, e.g., [4+2 ]]、[3+2]Etc. In some embodiments, the first and second reactive groups are two reactive groups suitable for cycloaddition reactions. In some embodiments, the reaction is a click reaction. In some embodiments, one reactive group is-N 3 Or comprises-N 3 And the other is or comprises an alkyne, e.g., a terminal alkyne or an activated/strained alkyne. In some embodiments, the other is or comprises-C.ident.CH.
In some embodiments, R of the first amino acid residue SP1 Or R is SP2 And R of a second amino acid residue SP1 Or R is SP2 Can react with the reagent such that the two join to form a staple-like structure. In some embodiments, the reagent comprises two reactive groups, one of which is R to the first amino acid residue SP1 Or R is SP2 And the other is reacted with R of the first amino acid residue SP1 Or R is SP2 And (3) reacting. In some embodiments, R of two amino acid residues SP1 Or R is SP2 Are of the same or the same type, e.g. both are amino groups, and the two reactive groups of the linking agent are also the same, e.g. both are acidic groups, e.g. -COOH or an activated form thereof. In some embodiments, R of two amino acid residues SP1 Or R is SP2 Are both acidic groups, such as-COOH or an activated form thereof, and both reactive groups of the linking reagent are amino groups. In some embodiments, R of two amino acid residues SP1 Or R is SP2 Are both nucleophilic groups, such as-SH, and both reactive groups of the linking reagent are electrophilic (e.g., carbon linked to a leaving group such as-Br, -I, etc.).
In some embodiments, R SP1 And R is SP2 Are identical. In some embodiments, R SP1 And R is SP2 Is different. In some embodiments, R SP1 is-CH=CH 2 Or comprises-CH=CH 2 . In some embodiments, R SP1 Is or comprises-COOH. In some embodiments, R SP1 Is or comprises an amino group. In some embodiments, R SP1 is-NHR or comprises-NHR. In some embodiments, R is hydrogen or optionally substituted C 1-6 Aliphatic series. In some embodiments, R SP1 is-NH 2 Or comprises-NH 2 . In some embodiments, R SP1 is-N 3 Or comprises-N 3 . In some embodiments, R SP2 is-CH=CH 2 Or comprises-CH=CH 2 . In some embodiments, R SP2 Is or comprises-COOH. In some embodiments, R SP2 Is amino or bagContains amino groups. In some embodiments, R SP2 is-NHR or comprises-NHR. In some embodiments, R is hydrogen or optionally substituted C 1-6 Aliphatic series. In some embodiments, R SP2 is-NH 2 Or comprises-NH 2 . In some embodiments, R SP2 is-N 3 Or comprises-N 3 。
In some embodiments, each amino acid residue in the pair of amino acid residues is independently a residue of an amino acid of formula A-II or A-III or a salt thereof. In some embodiments, such pairs of amino acid residues are stapled, for example, by olefin metathesis. In some embodiments, the staple-like structure has-L a -CH＝CH-L a -structure wherein each variable is independently as described herein. In some embodiments, the olefin in the staple-like structure is reduced. In some embodiments, the staple-like structure has a shape of-L a -CH 2 -CH 2 -L a -structure wherein each variable is independently as described herein. In some embodiments, one L a Is L as described herein s1 And one L a Is L as described herein s3 。
In some embodiments, for example, two amino acid residues, independent of the amino acids of formulas A-I or salts thereof, joined by a staple-like structure, have the formula-N (R a1 )-L a1 -C(-L s -R AA )(R a3 )-L a2 -CO-structure, wherein each variable is independently as described herein, and R AA Is an amino acid residue. In some embodiments, for example, two amino acid residues, independent of the amino acids of formulas A-I or salts thereof, joined by a staple-like structure, have a formula of-N (-L) s -R AA )-L a1 -C(R a2 )(R a3 )-L a1 -CO-structure, wherein each variable is independently as described herein, and R AA Is an amino acid residue. In some embodiments, for example, two amino acid residues, independent of the amino acids of formulas A-I or salts thereof, joined by a staple-like structure, have R a1 -N(-L s -R AA )-L a1 -C(R a2 )(R a3 )-L a2 -CO-structure, wherein each variable is independently as described herein, and R AA Is an amino acid residue. In some embodiments, for example, three amino acid residues, independent of the amino acids of formulas A-I or salts thereof, joined by two staple-like structures, have R a1 -N(-L s -R AA )-L a1 -C(-L s -R AA )(R a 3)-L a2 -CO-structure, wherein each variable is independently as described herein, and R AA Is an amino acid residue. In some embodiments, for example, three amino acid residues, independent of the amino acids of formulas A-I or salts thereof, joined by two staple-like structures, have a formula of-N (-L) s -R AA )-L a1 -C(-L s -R AA )(R a3 )-L a2 -CO-structure, wherein each variable is independently as described herein, and R AA Is an amino acid residue. In some embodiments, for example, by two staple-like structures (e.g., X 4 And X is 1 And X 14 Two stapled) linked three amino acid residues independent of the amino acids of formula A-I or salts thereof, having the formula-N (R a1 )-L a1 -C(-L s -R AA )(-L s -R AA )-L a2 -CO-structure, wherein each variable is independently as described herein, and R AA Is an amino acid residue. In some embodiments, each R AA Independently are residues of amino acids of the formulae A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or salts thereof. In some embodiments, R AA is-C (R) a3 )[-L a1 -N(R a1 )-](-L a2 -CO-), wherein each variable is independently as described herein. In some embodiments, R AA is-C (R) a3 )[-N(R a1 )-](-CO-) wherein each variable is independently as described herein. In some embodiments, each R AA Independently is-N (-) [ -L a1 -C(R a2 )(R a3 )-L a2 -CO-]Wherein each variable is independently as described herein, wherein-C (-) (R a3 ) -bonding with a staple-like structure. In some embodiments, each R AA Is independently-N (-) [ -C (R) a2 )(R a3 )-CO-]Wherein each variable is independently as described herein, wherein-C (-) (R a3 ) -bonding with a staple-like structure. In some embodiments, each R AA Independently is R a1 -N(-)[-L a1 -C(R a2 )(R a3 )-L a2 -CO-]Wherein each variable is independently as described herein, wherein-C (-) (R a3 ) -bonding with a staple-like structure. In some embodiments, each R AA Independently is R a1 -N(-)[-C(R a2 )(R a3 )-CO-]Wherein each variable is independently as described herein, wherein-C (-) (R a3 ) -bonding with a staple-like structure.
Staple-like structures, e.g. L s As described herein. In some embodiments, L s is-L s1 -L s2 -L s3 -, as described herein. In some embodiments, L s1 Is L a As described herein. In some embodiments, L s3 Is L a As described herein. In some embodiments, L s1 L being the first of two stapled amino acid residues a . In some embodiments, L s2 L being the second of two stapled amino acid residues a . In some embodiments, L s2 Is or comprises a double bond. In some embodiments, L s2 Is-ch=ch-or comprises-ch=ch-. In some embodiments, L s2 Is optionally substituted-CH 2 -CH 2 -or comprises optionally substituted-CH 2 -CH 2 -. In some embodiments, L s2 is-CH 2 -CH 2 -or comprise-CH 2 -CH 2 -. In some embodiments, L s2 is-C (O) N (R ') -, or comprises-C (O) N (R') - (e.g., a staple-like structure formed by two amino acid residues, one of which has R being or comprising an amino group) SP1 A group, and the other of which has an R which is or comprises-COOH SP2 A group). In some embodiments, L s2 is-C (O) NH-or contains-C (O) NH-. In some embodimentsWherein L is s1 And L s3 Each independently is an optionally substituted straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L s1 And L s3 Each independently is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L s1 is-CH 2 -. In some embodiments, L s3 Is- (CH) 2 ) 3 -。
In some embodiments, L s is-CH 2 -CH＝CH-(CH 2 ) 3 -. In some embodiments, L s Is- (CH) 2 ) 6 -。
In some embodiments, L s Is- (CH) 2 ) 2 -C(O)NH-(CH 2 ) 4 -。
In some embodiments, L s Bonded to two backbone carbon atoms. In some embodiments, L s Bonded to two alpha carbon atoms of two stapled amino acid residues. In some embodiments, L s Bonded to a backbone nitrogen atom and a backbone carbon atom (e.g., alpha carbon).
In some embodiments, L a Comprising at least one-N (R ') -wherein R' is independently as described in the present disclosure. In some embodiments, L a comprises-L am1 -N (R ') -, wherein R' is independently as described in the present disclosure, and L am1 As described herein. In some embodiments, L a is-L am1 -N(R’)-L am2 -or comprise-L am1 -N(R’)-L am2 -, wherein L am1 R' and L am2 Each independently as described herein. In some embodiments, R' is optionally substituted C 1-6 Aliphatic series. In some embodiments, R' is methyl. In some embodiments, R' and R a3 Taken together form an optionally substituted ring as described herein. In some embodiments, the ring formed is a 3-to 10-membered monocyclic saturated ring as described herein. In some embodiments, the formed ring has no additional heteroatom ring atoms other than the nitrogen of-N (R') -. In some embodiments, the ring formed is 3 And (3) meta. In some embodiments, the ring formed is 4-membered. In some embodiments, the ring formed is 5-membered. In some embodiments, the ring formed is 6 membered.
In some embodiments, L a Comprising at least one-C (R') 2 -, wherein each R' is independently as described in the present disclosure. In some embodiments, L a comprises-L am1 -C(R’) 2 -, wherein R' is independently as described in the disclosure, and L am1 As described herein. In some embodiments, L a is-L am1 -C(R’) 2 -L am2 -or comprise-L am1 -C(R’) 2 -L am2 -, wherein L am1 R' and L am2 Each independently as described herein. In some embodiments, R' is-H. In some embodiments, -C (R') 2 -is optionally substituted-CH 2 -. In some embodiments, -C (R') 2 -is-CH 2 -. In some embodiments, one R' and R a3 Taken together form an optionally substituted ring as described herein. In some embodiments, the ring formed is a 3-to 10-membered monocyclic saturated ring as described herein. In some embodiments, the formed ring has no additional heteroatom ring atoms other than the nitrogen of-N (R') -. In some embodiments, the ring formed is 3-membered. In some embodiments, the ring formed is 4-membered. In some embodiments, the ring formed is 5-membered. In some embodiments, the ring formed is 6 membered.
L as described herein am1 And L am2 Each independently is L am As described herein. L as described herein am Is a covalent bond or an optionally substituted divalent C 1 -C 10 Aliphatic groups wherein one or more methylene units of the aliphatic group are optionally and independently substituted with-C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R')-, -C (O) S-or-C (O) O-. At the position ofIn some embodiments, L am Is a covalent bond. In some embodiments, L am Is an optionally substituted divalent C 1 -C 10 An aliphatic group. In some embodiments, L am Is an optionally substituted divalent straight chain C 1 -C 10 An aliphatic group. In some embodiments, L am Is optionally substituted C 1-10 An alkylene group. In some embodiments, L am Is C 1-10 An alkylene group. In some embodiments, L am Is an optionally substituted straight chain C 1-10 An alkylene group. In some embodiments, L am Is optionally substituted-CH 2 -. In some embodiments, L am is-CH 2 -。
In some embodiments, L am1 Is a covalent bond. In some embodiments, L am1 Is an optionally substituted divalent C 1 -C 10 An aliphatic group. In some embodiments, L am1 Is an optionally substituted divalent straight chain C 1 -C 10 An aliphatic group. In some embodiments, L am1 Is optionally substituted C 1-10 An alkylene group. In some embodiments, L am1 Is C 1-10 An alkylene group. In some embodiments, L am1 Is an optionally substituted straight chain C 1-10 An alkylene group. In some embodiments, L am1 Is optionally substituted-CH 2 -. In some embodiments, L am1 is-CH 2 -. In some embodiments, L am1 Bonded to the backbone atoms. In some embodiments, L am1 Bonded to the alpha carbon of the amino acid.
In some embodiments, L am2 Is a covalent bond. In some embodiments, L am2 Is an optionally substituted divalent C 1 -C 10 An aliphatic group. In some embodiments, L am2 Is an optionally substituted divalent straight chain C 1 -C 10 An aliphatic group. In some embodiments, L am2 Is optionally substituted C 1-10 An alkylene group. In some embodiments, L am2 Is C 1-10 Alkylene groupA base. In some embodiments, L am2 Is an optionally substituted straight chain C 1-10 An alkylene group. In some embodiments, L am2 Is optionally substituted-CH 2 -. In some embodiments, L am2 is-CH 2 -. In some embodiments, L am2 is-C (O) -or comprises-C (O) -. In some embodiments, -C (O) -is bonded to a nitrogen atom. In some embodiments, L am2 is-S (O) 2 Or comprise-S (O) 2 -. In some embodiments, -S (O) 2 -bonding to a nitrogen atom. In some embodiments, L am2 is-O-or comprises-O-. In some embodiments, L am2 is-C (O) -O-or comprising-C (O) -O-. In some embodiments, -C (O) -O-is bonded to a nitrogen atom. In some embodiments, L am2 Bonded to nitrogen atoms, and L am2 comprising-C (O) -groups bonded to nitrogen atoms. In some embodiments, L am2 Bonded to nitrogen atoms, and L am2 Comprising a-C (O) -O-group bonded to a nitrogen atom. In some embodiments, L am2 is-C (O) -O-CH 2 -or comprises-C (O) -O-CH 2 -, wherein-CH 2 Optionally substituted. In some embodiments, L am2 is-C (O) -O-CH 2 -。
In some embodiments, L a Is L s1 As described herein. In some embodiments, L a Is L s2 As described herein.
In some embodiments, R a3 is-L a -CH＝CH 2 Wherein L is a Independently as described herein. In some embodiments, R a2 And R is a3 Each independently comprising a double bond, e.g., a terminal olefin that can be optionally and independently stapled with another residue comprising an olefin. In some embodiments, R a2 And R is a3 Each independently is-L a -CH＝CH 2 . In some embodiments, the amino acid is represented by R a2 And R is a3 Independently stapled with two amino acid residues. In some embodiments, such an amino acid is B5. In some embodiments, it Is B3. In some embodiments, it is B4. In some embodiments, it is B6.
In some embodiments, the amino acid is selected from tables A-I, A-II, A-III, and A-IV (which may be presented as Fmoc protected). As will be appreciated by those skilled in the art, particularly when incorporated into peptides, fmoc-protected amino and carboxyl groups can independently form amide linkages with other amino acid residues (either N-or C-terminal capping groups, or exist as N-or C-terminal amino or carboxyl groups). Olefins, including those in Alloc groups, can form staple-like structures by olefin metathesis. Staple-like structures comprising olefins may be further modified, for example by conversion of olefinic double bonds to single bonds by hydrogenation, and/or by CO 2 Extrusion converts a carbamate moiety (e.g., -O- (CO) -N (R ') (-) to an amine moiety (e.g., -N (R')) -). In some embodiments, the agent is or comprises a stapled peptide (e.g., a stapled peptide according to table E2 or table E3) or a salt thereof, wherein each double bond of the stapled peptide is converted to a single bond. In some embodiments, the conversion is achieved by hydrogenation by adding-H to each olefin carbon atom. In some embodiments, the olefinic double bond is replaced by-CHR '-wherein each R' is independently as described herein. In some embodiments, R' is R, as described herein. In some embodiments, R' is-H. In some embodiments, each R' is-H. In some embodiments, R' is-OR, wherein R is as described herein. In some embodiments, R' is-OH. In some embodiments, R' is-N (R) 2 Wherein each R is independently as described herein. In some embodiments, R' is-SR, wherein R is as described herein. In some embodiments, R' is R, where R is an optionally substituted aliphatic, e.g., C 1-10 Aliphatic series. In some embodiments, R' is R, where R is an optionally substituted aliphatic, e.g., C 1-10 Alkenyl groups. In some embodiments, R' is R, where R is an optionally substituted aliphatic, e.g., C 1-10 Alkynyl groups. In some embodiments, -CHR '-CHR' -is-CH 2 -CH 2 -。
Tables a-i. exemplary amino acids (Fmoc protected).
Tables a-ii. Exemplary amino acids (Fmoc protected).
Tables a-iii. Exemplary amino acids (Fmoc protected).
In some embodiments, the amino acid is an alpha-amino acid. In some embodiments, the amino acid is an L-amino acid. In some embodiments, the amino acid is a D-amino acid. In some embodiments, the alpha carbon of the amino acid is achiral. In some embodiments, the amino acid is a β -amino acid. In some embodiments, the amino acid is a gamma amino acid.
In some embodiments, provided amino acid sequences comprise two or more amino acid residues whose side chains are linked together to form one or more staple-like structures. In some embodiments, provided amino acid sequences comprise two or more amino acid residues, each independently having a side chain comprising an olefin. In some embodiments, provided amino acid sequences comprise two or more amino acid residues, each independently having a side chain comprising a terminal olefin. In some embodiments, provided amino acid sequences comprise two and no more than two amino acid residues, each independently having a side chain comprising an olefin. In some embodiments, the provided amino acid sequences comprise two and no more than two amino acid residues, which Each independently having a side chain comprising a terminal olefin. In some embodiments, provided amino acid sequences comprise at least one amino acid residue comprising an alkene and a nitrogen atom other than the nitrogen atom of its amino group. In some embodiments, provided amino acid sequences comprise at least one amino acid residue comprising a terminal alkene and a nitrogen atom other than the nitrogen atom of its amino group. In some embodiments, provided amino acid sequences comprise at least one amino acid residue having a side chain in addition to a terminal olefin and a nitrogen atom. In some embodiments, the provided amino acid sequences comprise at least one amino acid residue of formula A-I, wherein R a2 Comprising an alkene and a moiety-N (R ') -, wherein R' is as described in the present disclosure (in some embodiments, including optionally with R a3 And intervening atoms together to form an optionally substituted ring as described in this disclosure). In some embodiments, R a2 Comprising a terminal olefin and a moiety-N (R ') -, wherein R' is as described in the present disclosure. In some embodiments, the provided amino acid sequences comprise at least one amino acid residue selected from tables a-I. In some embodiments, the provided amino acid sequences comprise at least one amino acid residue selected from tables a-II. In some embodiments, the provided amino acid sequences comprise at least one amino acid residue selected from tables A-III. In some embodiments, the two olefins from the two side chains are linked together by olefin metathesis to form a staple-like structure. In some embodiments, the staple-like structure is preferably formed from side chains of amino acid residues that are not in the corresponding position of the target of interest. In some embodiments, the formed staple-like structure does not disrupt the interaction between the peptide and the target of interest.
In some embodiments, the staple-like structures provided are hydrocarbon staple-like structures. In some embodiments, the hydrocarbon staple-like structure does not contain chain heteroatoms, wherein the chain of the staple-like structure is the shortest covalent link within the staple-like structure from one end of the staple-like structure to the other end of the staple-like structure.
In some embodiments, the olefin in the staple-like structure is a Z-olefin. In some embodiments, the olefin in the staple-like structure is an E-olefin. In some embodiments, provided compositions comprise a stapled peptide comprising a staple-like structure comprising a Z-olefin, and a stapled peptide comprising a staple-like structure comprising an E-olefin. In some embodiments, provided compositions comprise a stapled peptide comprising a staple-like structure comprising a Z-olefin. In some embodiments, provided compositions comprise a stapled peptide comprising a staple-like structure comprising an E-olefin. In some embodiments, other aspects of the same stapled peptide that differ only in the E/Z configuration of the staple structured olefin exhibit different properties and/or activities as shown herein. In some embodiments, a stapled peptide containing an E-olefin in a staple-like structure may provide certain desirable properties and/or activities depending on the context. In some embodiments, a stapled peptide containing a Z-olefin in a staple-like structure may provide certain desirable properties and/or activities depending on the context.
In some embodiments, the present disclosure provides compositions comprising a stapled peptide. In some embodiments, the composition comprises one and only one stereoisomer (e.g., E or Z isomer, and/or a single diastereomer/enantiomer relative to a chiral center, etc.) of the stapled peptide. In some embodiments, the composition comprises two or more stereoisomers (e.g., both E and Z isomers of one or more double bonds, and/or one or more diastereomers/enantiomers relative to the chiral center, etc.). In some embodiments, the composition corresponds to a single peak in a chromatographic separation (e.g., HPLC). In some embodiments, the peak comprises one and only one stereoisomer. In some embodiments, the peak comprises two or more stereoisomers.
In some embodiments, two staple-like structures may bond to the same atom of the peptide backbone, forming a stitched peptide.
In some embodiments, the staple-like structure is pro-lock, wherein one end of the staple-like structure is bonded to the alpha carbon of the proline residue.
In some embodiments, the staple-like structures are those shown below in tables S-1, S-2, S-3, S-4, and S-5 (where exemplary peptide backbones (applicable to other peptide backbones) are shown for clarity, each X is independently an amino acid residue). In some embodiments, the staple-like structure is a staple-like structure in table S-6 (wherein amino acid residues bonded to the staple-like structure are shown). In some embodiments, the olefin is Z. In some embodiments, the olefin is E. In some embodiments, (i, i+3) staple like structures are selected from table S-1. In some embodiments, (i, i+3) staple like structures are selected from table S-2. Those skilled in the art who review this disclosure will appreciate that when the staple-like structures in tables S-1 and S-2 are used for (i, i+3), "X" in those tables 3 "will be" X 2 "(i.e., two amino acid residues instead of three amino acid residues). In some embodiments, (i, i+4) staple like structures are selected from table S-1. In some embodiments, (i, i+4) staple like structure is selected from table S-2. In some embodiments, (i, i+7) staple like structures are selected from table S-3. In some embodiments, (i, i+7) staple like structures are selected from table S-4.
Table S-1. Exemplary staple-like structures.
Table S-2. Exemplary staple-like structures.
Table S-3. Exemplary staple-like structures.
Table S-4. Exemplary staple-like structures.
Some useful staple-like structures are described in, for example, WO 2019/051327, WO 2022/020652, etc., and incorporated herein by reference.
In some embodiments, the staple-like structure may be one of the following, with amino acids attached at indicated positions:
TABLE S-5 certain amino acids and staple-like structures
In some embodiments, the peptide comprises a staple-like structure or a stitch-like structure (two staple-like structures) from table S-6. In Table 6, the amino acid residues may be from N to C or from C to N. In some embodiments, it is N to C. In some embodiments, it is C to N. In some embodiments, the double bond is E. In some embodiments, the double bond is Z. In some embodiments, the staple-like structure is an (i, i+2) staple-like structure. In some embodiments, the staple-like structure is an (i, i+3) staple-like structure. In some embodiments, the staple-like structure is an (i, i+4) staple-like structure. In some embodiments, the staple-like structure is an (i, i+7) staple-like structure. In some embodiments, when the structure comprises more than one double bond, each double bond is independently E or Z. In some embodiments, each staple-like structure is independently an (i, i+2) or an (i, i+3) or an (i, i+4) staple-like structure or an (i, i+7) staple-like structure. In some embodiments, each staple-like structure is independently an (i, i+2) or (i, i+4) staple-like structure or an (i, i+7) staple-like structure. In some embodiments, each staple-like structure is independently an (i, i+3) or (i, i+4) staple-like structure or an (i, i+7) staple-like structure. In some embodiments, in a structure comprising two staple-like structures, each staple-like structure is independently an (i, i+4) staple-like structure or an (i, i+7) staple-like structure. In some embodiments, one staple-like structure is an (i, i+4) staple-like structure and the other is an (i, i+7) staple-like structure. In some embodiments, one staple-like structure is an (i, i+3) staple-like structure, one staple-like structure is an (i, i+4) staple-like structure, and one staple-like structure is an (i, i+7) staple-like structure. In some embodiments, one staple-like structure is an (i, i+2) staple-like structure, one staple-like structure is an (i, i+4) staple-like structure, and one staple-like structure is an (i, i+7) staple-like structure. In some embodiments, the PL3 residue is bonded to an (i, i+3) staple-like structure. In some embodiments, the PL3 residue is bonded to an (i, i+4) staple-like structure. A kind of electronic device, PL3 and S5, R5 and S5, PL3 and B4 and identical B4 and PyrS1, PL3 and B4 and identical B4 and PyrS2, PL3 and B4 and identical B4 and PyrS3, PL3 and S6, PL3 and S4, PL3 and S3, R6 and PyrS2, R4 and PyrS2, R3 and PyrS2, PL3 and B3 and identical B3 and PyrS3, PL3 and B3 and identical B3 and PyrS4, PL3 and B6 and identical B6 and PyrS1, PL3 and B6 and identical B6 and PyrS2.
Table S-6. Certain staple-like structures (comprising amino acid residues bonded to the staple-like structure).
In some embodiments, the double bond in the (i, i+3) staple-like structure is Z. In some embodiments, the double bond in the (i, i+4) staple-like structure is Z. In some embodiments, the double bond in the (i, i+7) staple-like structure is Z. In some embodiments, the double bond in the (i, i+3) staple-like structure is E. In some embodiments, the double bond in the (i, i+4) staple-like structure is E. In some embodiments, the double bond in the (i, i+7) staple-like structure is E.
In some embodiments, the staple-like structure comprises-S-. In some embodiments, the stapling technique includes utilizing one or more, e.g., two or more sulfur-containing moieties. In some embodiments, the stapled peptide comprises cysteine stapling. In some embodiments, the two cysteine residues are stapled, wherein the-S-portions of the two cysteine residues are optionally connected by a linker. In some embodiments, the stapled peptide comprises one and no more than one staple-like structure from cysteine stapling. In some embodiments, the stapled peptide comprises one and no more than one staple-like structure having the structure: In some embodiments, the stapled peptide comprises one and no more than one staple-like structure having the structure:In some embodiments, the stapled peptide comprises one and no more than one staple-like structure having the structure:In some embodiments, the stapled peptide comprises one and no more than one staple-like structure having the structure:In some embodiments, the stapled peptide does not comprise a staple-like structure having the structure:In some embodiments, the stapled peptide does not comprise a staple-like structure having the structure:in some embodiments, the stapled peptide does not comprise a staple-like structure having the structure:In some embodiments, the stapled peptide does not comprise a staple-like structure having the structure:
In some embodiments, the present disclosure provides useful techniques related to cysteine stapling. The present disclosure contemplates, among other things, that peptides suitable for cysteine stapling and/or comprising one or more cysteine staple-like structures may be produced and/or evaluated in a biological system. The present disclosure also contemplates that certain such systems allow for the development, generation, and/or evaluation of cysteine stapled peptides having a range of different structures (e.g., different amino acid sequences), and in fact may provide the user with complete control over the selection and implementation of amino acid sequences to be incorporated into the stapled peptides.
As described herein, cysteine stapling involves linking one cysteine residue to another cysteine residue, wherein the resulting bond is not through the peptide backbone between the linked cysteine residues.
In some embodiments, a stapled peptide as described herein comprises a staple-like structure that is L s Wherein:
L s is-L s1 -S-L s2 -S-L s3 -；
L s1 And L s3 Each independently is L;
L s2 is L and comprises at least one-C (O) -; and is also provided with
Each L is independently a covalent bond or an optionally substituted divalent C 1 -C 25 An aliphatic group, wherein one or more methylene units of the aliphatic group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R')-, -C (O) S-or-C (O) O-;
each-Cy-is independently an optionally substituted divalent group selected from: c (C) 3-20 Cycloaliphatic ring, C 6-20 An aryl ring, a 5 to 20 membered heteroaryl ring having 1 to 10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus, and silicon, and a 3 to 20 membered heterocyclyl ring having 1 to 10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus, and silicon;
each R' is independently-R, -C (O) R, -CO 2 R or-SO 2 R；
Each R is independently-H, or an optionally substituted group selected from: c (C) 1-30 Aliphatic, C having 1 to 10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon 1-30 Heteroaliphatic, C 6-30 Aryl, C 6-30 Arylaliphatics, C having 1 to 10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon 6-30 Aryl heteroaliphatics, 5 to 30 membered heteroaryl groups having 1 to 10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus, and silicon, and 3 to 30 membered heterocyclyl groups having 1 to 10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus, and silicon, or
The two R groups optionally and independently taken together form a covalent bond, or
Two or more R groups on the same atom optionally and independently taken together with the atom form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having from 0 to 10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon in addition to the atom; or alternatively
Two or more R groups on two or more atoms optionally and independently together with their intervening atoms form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon in addition to the intervening atoms.
In some embodiments, L is independently divalent C 1 -C 25 An aliphatic group. In some embodiments, L is independently divalent C 1 -C 20 An aliphatic group. In some embodiments, L is independently divalent C 1 -C 10 An aliphatic group. In some embodiments, L is independently divalent C 1 -C 5 An aliphatic group. In some embodiments, L is independently divalent C 1 An aliphatic group. In some embodiments, L is-CH 2 。
In some embodiments, L s1 is-CH 2 -. In some embodiments, L s3 is-CH 2 -. In some embodiments, L s1 And L s3 Are all-CH 2 -. In some embodiments, L s is-CH 2 -S-L s2 -S-CH 2 -。
In some embodiments, L s2 comprising-C (R') 2 -L’-C(R’) 2 -, wherein L' is described in the present disclosure. In some embodiments, L s2 is-L x1 -C(O)Q-L’-QC(O)-L x1 -wherein each variable is independently as described in the present disclosure. In some embodiments, L s2 is-CH 2 C(O)Q-L’-QC(O)CH 2 -, wherein each-CH 2 -are independent and optionally substituted. In some embodiments, L s2 is-CH 2 C(O)Q-L’-QC(O)CH 2 -。
In some embodiments, L s2 In some embodiments, L s2 Is L and comprises at least one-C (O) -. In some embodiments, L s2 Is L and comprises at least two-C (O) -. In some embodiments, L s2 Is L and comprises at least one-C (O) Q-, whichWherein Q is selected from: covalent bond, -N (R'), -O-, and-S-. In some embodiments, L s2 Is L and comprises at least one-C (O) Q-, wherein Q is selected from the group consisting of-N (R') -and-O-. In some embodiments, L s2 Is L and comprises at least two-C (O) Q-, wherein Q is selected from: -N (R'), -O-and-S-. In some embodiments, L s2 Is L and comprises at least two-C (O) Q-, wherein Q is selected from the group consisting of-N (R') -and-O-. In some embodiments, L s2 Is L and comprises at least one-C (O) N (R') -. In some embodiments, L s2 Is L and comprises at least two-C (O) N (R') -. In some embodiments, L s2 Is L and comprises at least one-C (O) O-. In some embodiments, L s2 Is L and comprises at least two-C (O) O-.
In some embodiments, L s2 comprising-Q-L' -Q-, wherein Q is independently selected from: -N (R '), -O-and-S, wherein L' is described in the present disclosure.
In some embodiments, L s2 comprising-Q-L ' -Q-, wherein Q is independently selected from the group consisting of-N (R ') -and-O-, wherein L ' is described in the present disclosure. In some embodiments, L s2 comprising-C (O) Q-L' -QC (O) -, wherein Q is independently selected from: -N (R '), -O-and-S, wherein L' is described in the present disclosure. In some embodiments, L s2 comprising-C (O) Q-L ' -QC (O) -, wherein Q is independently selected from the group consisting of-N (R ') -and-O, wherein L ' is described in the present disclosure. In some embodiments, L s2 comprising-C (R') 2 C(O)Q-L’-QC(O)C(R’) 2 -, wherein Q is independently selected from: -N (R '), -O-and-S, wherein L' is described in the present disclosure. In some embodiments, L s2 comprising-C (R') 2 C(O)Q-L’-QC(O)C(R’) 2 -wherein Q is independently selected from-N (R ') -and-O, wherein L' is described in the present disclosure.
In some embodiments, L s2 comprising-N (R ') -L' -N (R ') -wherein L' is described in the present disclosure. In some embodiments, L s2 comprising-C (O) N (R ') -L' -N (R ') C (O) -, wherein L' is described in the present disclosure. In some embodiments，L s2 is-C (R') 2 C(O)N(R’)-L’-N(R’)C(O)C(R’) 2 -, wherein L' is described in the present disclosure.
In some embodiments, L s2 comprising-O (R ') -L' -O (R ') -wherein L' is described in the present disclosure. In some embodiments, L s2 comprising-C (O) O-L '-OC (O) -, wherein L' is described in the present disclosure. In some embodiments, L s2 is-C (R') 2 C(O)O-L’-OC(O)C(R’) 2 -, wherein L' is described in the present disclosure.
In some embodiments, R' is optionally substituted C 1-30 Aliphatic series. In some embodiments, R' is optionally substituted C 1-15 Aliphatic series. In some embodiments, R' is optionally substituted C 1-10 Aliphatic series. In some embodiments, R' is optionally substituted C 1-5 Aliphatic series. In some embodiments, R' is hydrogen.
In some embodiments, L' is an optionally substituted divalent C 1 -C 19 Aliphatic series. In some embodiments, L' is an optionally substituted divalent C 1 -C 15 Aliphatic series. In some embodiments, L' is an optionally substituted divalent C 1 -C 10 Aliphatic series. In some embodiments, L' is an optionally substituted divalent C 1 -C 9 Aliphatic series. In some embodiments, L' is an optionally substituted divalent C 1 -C 8 Aliphatic series. In some embodiments, L' is an optionally substituted divalent C 1 -C 7 Aliphatic series. In some embodiments, L' is an optionally substituted divalent C 1 -C 6 Aliphatic series. In some embodiments, L' is an optionally substituted divalent C 1 -C 5 Aliphatic series. In some embodiments, L' is an optionally substituted divalent C 1 -C 3 Aliphatic series. In some embodiments, L' is an optionally substituted divalent C 1 -C 2 Aliphatic series. In some embodiments, L' is an optionally substituted divalent C 1 Aliphatic series. In some embodiments, L'is-CH 2 -. In some embodiments, L' is- (CH) 2 ) 2 -. In some embodiments, L' is- (CH) 2 ) 3 -. In some embodiments, L' is- (CH) 2 ) 4 -. In some embodiments, L' is- (CH) 2 ) 5 -. In some embodiments, L' is- (CH) 2 ) 6 -. In some embodiments, L' is- (CH) 2 ) 7 -. In some embodiments, L' is- (CH) 2 ) 8 -。
In some embodiments, L' is an optionally substituted divalent C 6-20 An aryl ring. In some embodiments, L' is an optionally substituted divalent C 6-14 An aryl ring. In some embodiments, L' is an optionally substituted divalent C 6-10 An aryl ring. In some embodiments, L' is an optionally substituted divalent C 6 An aryl ring. In some embodiments, L' is a divalent C substituted with at least one halogen 6 Aryl groups. In some embodiments, L' is a divalent C substituted with at least two halogens 6 Aryl groups. In some embodiments, L' is a divalent C substituted with at least three halogens 6 Aryl groups. In some embodiments, L' is a divalent C substituted with four halogens 6 Aryl groups. In some embodiments, L' is a divalent C substituted with at least one fluorine 6 Aryl groups. In some embodiments, L' is a divalent C substituted with at least two fluorine groups 6 Aryl groups. In some embodiments, L' is a divalent C substituted with at least three fluorine groups 6 Aryl groups. In some embodiments, L' is a divalent C substituted with four fluorine groups 6 Aryl groups. In some embodiments, L' is a divalent C substituted with at least one chlorine 6 Aryl groups. In some embodiments, L' is a divalent C substituted with at least two chlorines 6 Aryl groups. In some embodiments, L' is a divalent C substituted with at least three chlorides 6 Aryl groups. In some embodiments, L' is a divalent C substituted with four chlorides 6 Aryl groups. In some embodiments, L' is a compound substituted with at least one-O (CH 2 ) 0-4 CH 3 Substituted divalent C 6 Aryl groups. In some embodiments, L' is a compound containing at least two groups of-O (CH 2 ) 0-4 CH 3 Substituted divalent C 6 Aryl groups. In some embodiments, L' is a compound substituted with at least three-O (CH 2 ) 0-4 CH 3 Substituted divalent C 6 Aryl groups. In some embodiments, L' is a single-unit formed by four-O (CH 2 ) 0-4 CH 3 Substituted divalent C 6 Aryl groups.
In some embodiments, L' is a divalent 5-to 20-membered heteroaryl ring having 1 to 10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus, and silicon. In some embodiments, L' is a divalent 5-to 6-membered heteroaryl ring having 1 to 4 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus, and silicon. In some embodiments, L' is a divalent 5-to 6-membered heteroaryl ring having 1 to 4 heteroatoms independently selected from oxygen, nitrogen, and sulfur. In some embodiments, L' is a divalent 6 membered heteroaryl ring having 1 to 2 heteroatoms independently selected from oxygen, nitrogen, and sulfur. In some embodiments, L' is a divalent 6 membered heteroaryl ring having 2 nitrogens.
In some embodiments, L' is an optionally substituted divalent C 3-20 Cycloaliphatic rings. In some embodiments, L' is an optionally substituted divalent C 3-15 Cycloaliphatic rings. In some embodiments, L' is an optionally substituted divalent C 3-10 Cycloaliphatic rings. In some embodiments, L' is an optionally substituted divalent C 3-9 Cycloaliphatic rings. In some embodiments, L' is an optionally substituted divalent C 3-8 Cycloaliphatic rings. In some embodiments, L' is an optionally substituted divalent C 3-7 Cycloaliphatic rings. In some embodiments, L' is an optionally substituted divalent C 3-6 Cycloaliphatic rings. In some embodiments, L' is an optionally substituted divalent C 3-5 Cycloaliphatic rings. In some embodiments, L' is an optionally substituted divalent C 3-4 Cycloaliphatic rings. In some embodiments, L' is an optionally substituted divalent C 3 Cycloaliphatic rings. In some embodiments, L' is an optionally substituted divalent C 4 Cycloaliphatic rings.In some embodiments, L' is an optionally substituted divalent C 5 Cycloaliphatic rings. In some embodiments, L' is an optionally substituted divalent C 5 Cycloalkyl rings. In some embodiments, L' is an optionally substituted divalent C 5 A cycloalkenyl ring. In some embodiments, L' is an optionally substituted divalent C 6 Cycloaliphatic rings. In some embodiments, L' is an optionally substituted divalent C 6 Cycloalkyl rings.
In some embodiments, L s2 comprises-N (R ') -L' -N (R ') -and L' is a covalent bond. In some embodiments, L s2 comprising-N (R) -, wherein:
each R is independently-H, or an optionally substituted group selected from: c (C) 1-30 Aliphatic, C having 1 to 10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon 1-30 Heteroaliphatic, C 6-30 Aryl, C 6-30 Arylaliphatics, C having 1 to 10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon 6-30 Aryl heteroaliphatics, 5 to 30 membered heteroaryl groups having 1 to 10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus, and silicon, and 3 to 30 membered heterocyclyl groups having 1 to 10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus, and silicon, or
Two or more R groups on two or more atoms optionally and independently together with their intervening atoms form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon in addition to the intervening atoms.
In some embodiments, L s2 comprising-N (R) -, wherein:
each R is independently optionally substituted C 1-30 Aliphatic series; or alternatively
Two or more R groups on two or more atoms optionally and independently taken together with intervening atoms form an optionally substituted 3-to 30-membered monocyclic ring.
In some embodiments, L s2 Is a staple-like structure selected from the group consisting of:
in some embodiments, L s1 Is an optionally substituted divalent C 1-6 Aliphatic series. In some embodiments, L s1 Is divalent C 1-6 Aliphatic series. In some embodiments, L s1 Is divalent C 1-4 Aliphatic series. In some embodiments, L s1 Is saturated. In some embodiments, L s1 Is linear. In some embodiments, L s1 Is branched. In some embodiments, L s1 Is optionally substituted-CH 2 -. In some embodiments, L s1 is-CH 2 -. In some embodiments, L s1 Is optionally substituted-CH 2 -CH 2 -. In some embodiments, L s1 is-CH 2 -CH 2 -. In some embodiments, L s1 Is optionally substituted-C (CH) 3 ) 2 -. In some embodiments, L s1 is-C (CH) 3 ) 2 -。
In some embodiments, L s2 Is an optionally substituted divalent C 1-6 (e.g., C 3-6 、C 3 、C 4 、C 5 、C 6 Etc.) aliphatic wherein one or more methylene units are optionally and independently substituted with-Cy-or-C (R') 2 -substitution. In some embodiments, L s2 Is an optionally substituted divalent C 1-6 Aliphatic series. In some embodiments, L s2 Is an optionally substituted divalent C 3-6 Aliphatic series. In some embodiments, L s2 Is divalent C 1-6 Aliphatic series. In some embodiments, L s2 Is divalent C 1-4 Aliphatic series. In some embodiments, L s2 Is an optionally substituted divalent C 2 Aliphatic series. In some embodiments, L s2 Is an optionally substituted divalent C 3 Aliphatic series. In some casesIn embodiments, L s2 Is an optionally substituted divalent C 4 Aliphatic series. In some embodiments, L s2 Is an optionally substituted divalent C 5 Aliphatic series. In some embodiments, L s2 Is an optionally substituted divalent C 6 Aliphatic series. In some embodiments, L s2 Is substituted. In some embodiments, L s2 Is unsubstituted. In some embodiments, L s2 Is saturated. In some embodiments, L s2 Is linear. In some embodiments, L s2 Is branched. In some embodiments, L s2 Is an optionally substituted divalent C 3-6 (e.g., C 3-5 、C 3 、C 4 、C 5 、C 6 Etc.) aliphatic, wherein one or both methylene units are independently replaced by-Cy-. In some embodiments, L s2 is-CH 2 -Cy-CH 2 -. In some embodiments, L s2 is-CH 2 -CH 2 -Cy-CH 2 -CH 2 -. In some embodiments, L s2 is-CH 2 -Cy-Cy-CH 2 -. Several useful embodiments of Cy-are as described herein. For example, in some embodiments, -Cy-is an optionally substituted monocyclic 5-membered aromatic ring having 0 to 4 heteroatoms. In some embodiments, -Cy-is an optionally substituted monocyclic 6-membered aromatic ring having 0 to 4 heteroatoms. In some embodiments, -Cy-is optionally substituted phenylene. In some embodiments, -Cy-is optionally substituted 1, 2-phenylene. In some embodiments, -Cy-is 1, 2-phenylene. In some embodiments, -Cy-is optionally substituted 1, 3-phenylene. In some embodiments, -Cy-is 1, 3-phenylene. In some embodiments, -Cy-is optionally substituted 1, 5-phenylene. In some embodiments, -Cy-is 1, 5-phenylene. In some embodiments, -Cy-is 3-methyl-1, 5-phenylene. In some embodiments, -Cy-is 3-methoxy-1, 5-phenylene. In some embodiments, -Cy-is an optionally substituted divalent pyridinyl ring. In some embodiments, -Cy-is optionally substitutedIn some embodiments, -Cy-is +.>In some embodiments, -Cy-is optionally substituted +. >In some embodiments, -Cy-is +.>In some embodiments, -Cy-is an optionally substituted bicyclic 9-membered aromatic ring having 0 to 4 heteroatoms. In some embodiments, -Cy-is an optionally substituted bicyclic 10-membered aromatic ring having 0 to 4 heteroatoms. In some embodiments, -Cy-is an optionally substituted divalent naphthyl ring. In some embodiments, -Cy-is a divalent naphthyl ring. In some embodiments, -Cy-is optionally substituted +.>In some embodiments, -Cy-is +.>In some embodiments, -Cy-is optionally substituted +.>In some embodiments, -Cy-is +.>In some embodiments, -Cy-is an optionally substituted 3 to 10 (e.g., 5 to 10, 5 to 6, 3, 4, 5, 6, 7, 8, 9, 10, etc.) membered divalent cycloaliphatic ring. In some embodiments, it is saturated. In some embodiments, -Cy-is an optionally substituted 6 membered cycloalkyl ring. In some embodiments, -Cy-is optionally substituted +.>In some embodiments, -Cy-is +.>In some embodiments, L s2 Is an optionally substituted divalent C 3-6 (e.g., C 3-5 、C 3 、C 4 、C 5 、C 6 Etc.) aliphatic wherein one or both methylene units are independently replaced by-C (R') 2 -substitution. In some embodiments, L s2 is-CH 2 -C(R’) 2 -CH 2 -. In some embodiments, two R' and a carbon atom are taken together to form an optionally substituted ring as described herein, e.g., an optionally substituted 3 to 10 (e.g., 5 to 10, 5 to 6, 3, 4, 5, 6, 7, 8, 9, 10, etc.) membered ring having 0 to 4 (e.g., 1 to 4, 0, 1, 2, 3, 4, etc.) heteroatoms. In some embodiments, the ring is saturated. In some embodiments, the ring has one or more heteroatoms. In some embodiments, -C (R') 2 -is->
In some embodiments, L s2 Is optionally substitutedIn some embodiments, L s2 Is optionally substituted->In some embodiments, L s2 Is optionally substituted->In some embodiments, L s2 Is optionally substituted->In some embodimentsIn the scheme, L s2 Is optionally substitutedIn some embodiments, L s2 Is optionally substituted->In some embodiments, L s2 Is optionally substituted->In some embodiments, L s2 Is optionally substituted->In some embodiments, L s2 Is optionally substituted->In some embodiments, L s2 Is optionally substitutedIn some embodiments, L s2 Is optionally substituted- (CH) 2 ) 4 -. In some embodiments, L s2 Is optionally substituted- (CH) 2 ) 3 -. In some embodiments, L s2 Is optionally substituted-CH 2 -CH＝CH-CH 2 -. In some embodiments, L s2 Is optionally substituted (E) -CH 2 -CH＝CH-CH 2 -. In some embodiments, L s2 Is optionally substituted-CH 2 -C(O)-CH 2 -. In some embodiments, L s2 Is optionally substituted->In some embodiments, L s2 Is optionally substituted->In some embodiments, L s2 Is optionally substituted->In some embodiments, L s2 Is optionally substituted->In some embodiments, it is substituted. In some embodiments, it is unsubstituted. In some embodiments, under comparable conditions, with +.> And/or +.>In contrast to this, the method comprises,-(CH 2 ) 4 -,(E)-CH 2 -CH＝CH-CH 2 -,-(CH 2 ) 3 -, and/or-CH 2 -C(O)-CH 2 Providing higher binding and/or efficacy.
In some embodiments, L s3 Is an optionally substituted divalent C 1-6 Aliphatic series. In some embodiments, L s3 Is divalent C 1-6 Aliphatic series. In some embodiments, L s3 Is divalent C 1-4 Aliphatic series. In some embodiments, L s3 Is saturated. In some embodiments, L s3 Is linear. In some embodiments, L s3 Is branched. In some embodiments, L s3 Is optionally substituted-CH 2 -. In some embodiments, L s3 is-CH 2 -. In some embodiments, L s3 Is optionally substituted-CH 2 -CH 2 -. In some embodimentsWherein L is s3 is-CH 2 -CH 2 -. In some embodiments, L s3 Is optionally substituted-C (CH) 3 ) 2 -. In some embodiments, L s3 is-C (CH) 3 ) 2 -。
In some embodiments, the amino acid residues used to form the staple-like structure are selected from the group consisting of:
in some embodiments, the two amino acid residues used to form the staple-like structure are independently residues of these amino acids. In some embodiments, L s1 And L s3 Each independently is-CH 2 -、-CH 2 -CH 2 -or-C (CH) 3 ) 2 -. In some embodiments, the staple-like structure is formed by reacting a thiol group with a thiol-group reactive linker compound. In some embodiments, such linker compounds have LG-L s2 -LG structure or salt thereof, wherein each LG is independently a leaving group, e.g. -Br, -I, or the like. In some embodiments, each LG is independently-Br or-I. In some embodiments, each LG is-Br. In some embodiments, each LG is-I. In some embodiments, L s2 Has such a structure that LG-L s2 LG (each LG is independently-Br or-I) is a compound selected from the group consisting of:
A variety of techniques are available for constructing thioether staple like structures. For example, in some embodiments, the peptide and excess equivalents (e.g., about 2 to 10, 5 to 10, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.; in some embodiments, 5) of the linker compound are added to 1:1DMF:100mM Na 2 CO 3 The pH 8.0 solution, and stirred at a suitable temperature, e.g., room temperature, for a suitable period of time, in some embodiments 1 to 2 hours. In some embodiments, for example, for relatively weak electrophiles, an excess of equivalent (e.g., about 10 to 30, 10 to 20, 10, 20, etc.; in some embodiments, 20) of a metal salt such as Zn (acac) 2 And excess equivalents (e.g., about 5 to 20, 10 to 15, 10, 15, 20, etc.; in some embodiments, 10 to 15) of the linker compound to the peptide in DMA, and the mixture is stirred at a suitable temperature, e.g., 37 ℃ for a suitable period of time, e.g., overnight. In some embodiments, zn (acac) 2 And doubling the equivalent weight of the linker compound, and/or increasing the temperature to 50 ℃. In some embodiments, certain linker compounds react better than others. For example, in some embodiments, the first and second substrates,
providing poor reaction yields or ineffective reactions. Those skilled in the art will appreciate that other techniques may be utilized to introduce the corresponding linker moiety (L s2 ) For example by using other leaving groups or by other reaction mechanisms/pathways.
In some embodiments, have-L s1 -S-L s2 -S-L s3 The staple-like structure of the structure is an (i, i+4) staple-like structure. In some embodiments, such staple-like structures are closer to the C-terminus. In some embodiments, such staple-like structures are closer to the N-terminus. For example, in some embodiments, such staple-like structures are at X 10 And X is 14 Between them.
In some embodiments, certain staple-like structures provide better properties and/or activity. For example, in some embodiments, certain staple-like structures/scaffolds are arranged in the following order based on target binding affinity:
as will be appreciated by those skilled in the art, the techniques provided may be used to prepare peptide sets using non-cysteine residues and their appropriate chemicals. For example, in some embodiments, cysteine stapling is replaced with lysine stapling, wherein the cysteine residues used for cysteine stapling are replaced with lysine residues used for lysine stapling (e.g., using a reagent that can crosslink two lysine residues, e.g., by reaction with a side chain amino group). In some embodiments, for lysine stapling, R in multiple formulas E Is or comprises an activated carboxylic acid group (e.g., NHS ester group), an imide ester group, etc. Suitable reagents are widely known in the art and include a number of commercially available reagents. In some embodiments, cysteine stapling is replaced with methionine stapling. In some embodiments, the cysteine residues used for cysteine stapling are replaced with methionine residues used for methionine stapling. In some embodiments, cysteine stapling is replaced with tryptophan stapling. In some embodiments, the cysteine residues used for cysteine stapling are replaced with tryptophan residues used for tryptophan stapling. As will be appreciated by those of skill in the art, a variety of techniques (e.g., reagents, reactions, etc.) are described in the art and may be used in accordance with the present disclosure, for example, methionine stapling, tryptophan stapling, etc. In some embodiments, such stapling can be performed using reagents having a variety of formulae described herein, wherein R E Is or comprises a group suitable for methionine and/or tryptophan stapling. In some embodiments, stapling can be performed using one residue in the first position and a different residue in the second position. The reagents available for such stapling may include a reagent for binding in a first position (e.g., by a first R E ) A first reactive group for stapling and for attaching to a second site (e.g., by a second R E ) StapledA second reactive group.
In some embodiments, for multiple types of stapling (e.g., cysteine stapling or non-cysteine stapling), stapling is between residues separated by two residues (e.g., cysteine residues for cysteine stapling) (i+3 stapling). In some embodiments, stapling is between residues separated by three residues (i+4 stapling). In some embodiments, stapling is between residues separated by six residues (i+7 stapling).
As will be appreciated by those skilled in the art, in some embodiments, more than two residues may be stapled simultaneously. For example, in some embodiments, a polymer comprising three or more reactive groups (e.g., R E A group) stapling three or more cysteines.
In some embodiments, the present disclosure provides useful techniques related to non-cysteine stapling, as described herein. The present disclosure contemplates, among other things, peptides suitable for cysteine stapling and/or comprising one or more non-cysteine staple-like structures, the cysteine residues and cysteine staple-like structures of which can be replaced with other amino acids and staple-like structures (e.g., hydrocarbons and other non-hydrocarbon amino acids and staple-like structures) described herein. In some embodiments, the resulting non-cysteine stapled peptide maintains the same or similar interaction with the target of interest when compared to the reference cysteine stapled peptide.
Certain useful agents (peptides prior to stapling and stapled peptides after stapling) and combinations thereof are shown as examples in table E2 or table E3, which include N-and C-terminal capping groups at different positions as examples, as well as a variety of amino acid residues; also shown are various stapling modes, e.g. X 1 -X 4 -X 11 ，X 1 -X 3 ，X 3 -X 7 ，X 3 -X 10 ，X 4 -X 11 ，X 7 -X 10 ，X 7 -X 14 ，X 10 -X 14 Etc. As demonstrated herein, the provided techniques may deliver improved useful properties and/or activity.
In some embodiments, the provided agent, peptide, or stapled peptide is a compound as described herein. In some embodiments, the provided agents have a structure selected from table E2 or table E3, or a salt thereof. In some embodiments, the provided agent is a stereoisomer selected from the structures of table E2 or E3, or a salt thereof. In some embodiments, the provided agent is a stereoisomer of a structure selected from table E2 or table E3, or a salt thereof, relative to the chiral center bonded to both staple-like structures (e.g., in B4, B5, etc.). In some embodiments, the provided agent is a stereoisomer of a structure selected from table E2 or table E3, or a salt thereof, relative to the olefinic double bond in the staple-like structure. In some embodiments, the provided agent is a stereoisomer of a structure selected from table E2 or E3, or a salt thereof, relative to the olefinic double bond in the staple-like structure and/or the chiral center bonded to both staple-like structures (e.g., in B4, B5, etc.). In some embodiments, the provided compositions are the compositions described in table E2 or table E3. In some embodiments, the compound has the following structure or salt thereof:
In some embodiments, the compound has the following structure or salt thereof:
in some embodiments, the compound has the following structure or salt thereof:
in some embodiments, the compound has the following structure or salt thereof:
in some embodiments, the compound has the following structure or salt thereof:
in some embodiments, the compound has the following structure or salt thereof:
in some embodiments, the compound has the following structure or salt thereof:
in some embodiments, the compound has the following structure or salt thereof:
in some embodiments, the compound has the following structure or salt thereof:
in some embodiments, the compound has the following structure or salt thereof:
in some embodiments, the compound has the following structure or salt thereof:
in some embodiments, the compound has the following structure or salt thereof:
in some embodiments, the compound has the following structure or salt thereof:
in some embodiments, the compound has the following structure or salt thereof:
in some embodiments, the compound has the following structure or salt thereof:
in some embodiments, the double bond of the (i, i+2) staple structure is E. In some embodiments, the double bond of the (i, i+2) staple structure is Z. In some embodiments, the double bond of the (i, i+3) staple-like structure is E. In some embodiments, the double bond of the (i, i+3) staple-like structure is Z. In some embodiments, the double bond of the (i, i+7) staple-like structure is E. In some embodiments, the double bond of the (i, i+7) staple-like structure is Z. In some embodiments, both double bonds are E. In some embodiments, both double bonds are Z. In some embodiments, (i, i+3) staple-like structure is E and the other is Z. In some embodiments, (i, i+3) staple-like structure is Z, and the other is E. In some embodiments, (i, i+4) staple-like structure is E and the other is Z. In some embodiments, (i, i+4) staple-like structure is Z, and the other is E. In some embodiments, the double bond of the (i, i+7) staple structure is Z, and the double bond of the second staple structure (e.g., (i, i+2), (i, i+3), (i, i+4), etc.) is E. In some embodiments, the double bond of the (i, i+7) staple structure is Z, and the double bond of the second staple structure (e.g., (i, i+2), (i, i+3), (i, i+4), etc.) is Z. In some embodiments, the double bond of the (i, i+7) staple-like structure is E, and the double bond of the second staple-like structure (e.g., (i, i+2), (i, i+3), (i, i+4), etc.) is E. In some embodiments, the double bond of the (i, i+7) staple-like structure is E, and the double bond of the second staple-like structure (e.g., (i, i+2), (i, i+3), (i, i+4), etc.) is Z. In some embodiments, the two staple-like structures are bonded to a chiral center (e.g., a carbon atom in B5), and the chiral center is R. In some embodiments, the two staple-like structures are bonded to a chiral center (e.g., a carbon atom in B5), and the chiral center is S.
In some embodiments, the compound has a structure selected from the group consisting of:
in some embodiments, the agent is SP-1-1 or a salt thereof. In some embodiments, the agent is SP-1-2 or a salt thereof. In some embodiments, the agent is SP-1-3 or a salt thereof. In some embodiments, the agent is SP-1-4 or a salt thereof. In some embodiments, the agent is SP-1-5 or a salt thereof. In some embodiments, the agent is SP-1-6 or a salt thereof. In some embodiments, the agent is SP-1-7 or a salt thereof. In some embodiments, the agent is SP-1-8 or a salt thereof. In some embodiments, the agent is SP-2-1 or a salt thereof. In some embodiments, the agent is SP-2-2 or a salt thereof. In some embodiments, the agent is SP-2-3 or a salt thereof. In some embodiments, the agent is SP-2-4 or a salt thereof. In some embodiments, the agent is SP-2-5 or a salt thereof. In some embodiments, the agent is SP-2-6 or a salt thereof. In some embodiments, the agent is SP-2-7 or a salt thereof. In some embodiments, the agent is SP-2-8 or a salt thereof. In some embodiments, the agent is SP-3-1 or a salt thereof. In some embodiments, the agent is SP-3-2 or a salt thereof. In some embodiments, the agent is SP-4-1 or a salt thereof. In some embodiments, the agent is SP-4-2 or a salt thereof. In some embodiments, the agent is SP-4-3 or a salt thereof. In some embodiments, the agent is SP-4-4 or a salt thereof. In some embodiments, the agent is SP-4-5 or a salt thereof. In some embodiments, the agent is SP-4-6 or a salt thereof. In some embodiments, the agent is SP-4-7 or a salt thereof. In some embodiments, the agent is SP-4-8 or a salt thereof. In some embodiments, the agent is SP-5-1 or a salt thereof. In some embodiments, the agent is SP-5-2 or a salt thereof. In some embodiments, the agent is SP-5-3 or a salt thereof. In some embodiments, the agent is SP-5-4 or a salt thereof. In some embodiments, the agent is SP-5-5 or a salt thereof. In some embodiments, the agent is SP-5-6 or a salt thereof. In some embodiments, the agent is SP-5-7 or a salt thereof. In some embodiments, the agent is SP-5-8 or a salt thereof. In some embodiments, the agent is SP-6 or a salt thereof. In some embodiments, the agent is SP-7-1 or a salt thereof. In some embodiments, the agent is SP-7-2 or a salt thereof. In some embodiments, the agent is SP-7-3 or a salt thereof. In some embodiments, the agent is SP-7-4 or a salt thereof. In some embodiments, the agent is SP-7-5 or a salt thereof. In some embodiments, the agent is SP-7-6 or a salt thereof. In some embodiments, the agent is SP-7-7 or a salt thereof. In some embodiments, the agent is SP-7-8 or a salt thereof. In some embodiments, the agent is SP-8-1 or a salt thereof. In some embodiments, the agent is SP-8-2 or a salt thereof. In some embodiments, the agent is SP-8-3 or a salt thereof. In some embodiments, the agent is SP-8-4 or a salt thereof. In some embodiments, the agent is SP-8-5 or a salt thereof. In some embodiments, the agent is SP-8-6 or a salt thereof. In some embodiments, the agent is SP-8-7 or a salt thereof. In some embodiments, the agent is SP-8-8 or a salt thereof. In some embodiments, the agent is SP-9-1 or a salt thereof. In some embodiments, the agent is SP-9-2 or a salt thereof. In some embodiments, the agent is SP-9-3 or a salt thereof. In some embodiments, the agent is SP-9-4 or a salt thereof. In some embodiments, the agent is SP-9-5 or a salt thereof. In some embodiments, the agent is SP-9-6 or a salt thereof. In some embodiments, the agent is SP-9-7 or a salt thereof. In some embodiments, the agent is SP-9-8 or a salt thereof. In some embodiments, the agent is SP-10-1 or a salt thereof. In some embodiments, the agent is SP-10-2 or a salt thereof. In some embodiments, the agent is SP-10-3 or a salt thereof. In some embodiments, the agent is SP-10-4 or a salt thereof. In some embodiments, the agent is SP-10-5 or a salt thereof. In some embodiments, the agent is SP-10-6 or a salt thereof. In some embodiments, the agent is SP-10-7 or a salt thereof. In some embodiments, the agent is SP-10-8 or a salt thereof. In some embodiments, the agent is SP-11-1 or a salt thereof. In some embodiments, the agent is SP-11-2 or a salt thereof. In some embodiments, the agent is SP-11-3 or a salt thereof. In some embodiments, the agent is SP-11-4 or a salt thereof. In some embodiments, the agent is SP-11-5 or a salt thereof. In some embodiments, the agent is SP-11-6 or a salt thereof. In some embodiments, the agent is SP-11-7 or a salt thereof. In some embodiments, the agent is SP-11-8 or a salt thereof. In some embodiments, the agent is SP-12-1 or a salt thereof. In some embodiments, the agent is SP-12-2 or a salt thereof. In some embodiments, the agent is SP-12-3 or a salt thereof. In some embodiments, the agent is SP-12-4 or a salt thereof. In some embodiments, the agent is SP-12-5 or a salt thereof. In some embodiments, the agent is SP-12-6 or a salt thereof. In some embodiments, the agent is SP-12-7 or a salt thereof. In some embodiments, the agent is SP-12-8 or a salt thereof. In some embodiments, the agent is SP-13-1 or a salt thereof. In some embodiments, the agent is SP-13-2 or a salt thereof. In some embodiments, the agent is SP-13-3 or a salt thereof. In some embodiments, the agent is SP-13-4 or a salt thereof. In some embodiments, the agent is SP-13-5 or a salt thereof. In some embodiments, the agent is SP-13-6 or a salt thereof. In some embodiments, the agent is SP-13-7 or a salt thereof. In some embodiments, the agent is SP-13-8 or a salt thereof. In some embodiments, the agent is SP-14-1 or a salt thereof. In some embodiments, the agent is SP-14-2 or a salt thereof. In some embodiments, the agent is SP-14-3 or a salt thereof. In some embodiments, the agent is SP-14-4 or a salt thereof. In some embodiments, the agent is SP-14-5 or a salt thereof. In some embodiments, the agent is SP-14-6 or a salt thereof. In some embodiments, the agent is SP-14-7 or a salt thereof. In some embodiments, the agent is SP-14-8 or a salt thereof. In some embodiments, the agent is SP-15-1 or a salt thereof. In some embodiments, the agent is SP-15-2 or a salt thereof. In some embodiments, the agent is SP-15-3 or a salt thereof. In some embodiments, the agent is SP-15-4 or a salt thereof. In some embodiments, the agent is SP-15-5 or a salt thereof. In some embodiments, the agent is SP-15-6 or a salt thereof. In some embodiments, the agent is SP-15-7 or a salt thereof. In some embodiments, the agent is SP-15-8 or a salt thereof.
Agents, such as peptides comprising stapled peptides, may comprise a different number of amino acid residues. In some embodiments, the peptide agent is about 5 to 20, 5 to 19, 5 to 18, 5 to 17, 5 to 16, 5 to 15, 10 to 20, 10 to 19, 10 to 18, 10 to 17, 10 to 16, 10 to 15, 11 to 20, 11 to 19, 11 to 18, 11 to 17, 11 to 16, 11 to 15, 12 to 20, 12 to 19, 12 to 18, 12 to 17, 12 to 16, 12 to 15, 13 to 20, 13 to 19, 13 to 18, 13 to 17, 13 to 16, 13 to 15, 14 to 20, 14 to 19, 14 to 18, 14 to 17, 14 to 16, 14 to 15, or about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acid residues in length. In some embodiments, the length is about 10 amino acid residues. In some embodiments, the length is about 11 amino acid residues. In some embodiments, the length is about 12 amino acid residues. In some embodiments, the length is about 13 amino acid residues. In some embodiments, the length is about 14 amino acid residues. In some embodiments, the length is about 15 amino acid residues. In some embodiments, the length is about 16 amino acid residues. In some embodiments, the length is about 17 amino acid residues. In some embodiments, the length is about 18 amino acid residues. In some embodiments, the length is about 19 amino acid residues. In some embodiments, the length is about 20 amino acid residues.
In some embodiments, as described herein, one or more staple-like structures independently comprise an olefinic double bond (e.g., formed by olefin metathesis). In some embodiments, one or more of the staple-like structures independently comprise an amide group (e.g., formed by amidation). In some embodiments, at least one staple-like structure does not contain olefinic double bonds. In some embodiments, the formation of at least one staple-like structure does not include an olefin reaction, such as olefin metathesis and/or modification of an olefin double bond (e.g., hydrogenation, epoxidation, etc.).
In some embodiments, the residues of the staple-like structure (e.g., B5) are positioned such that if their positions are P (e.g., X 4 ) Then the first acidic amino acid residue is at position P-2 (e.g., X 2 ) Second acidic amino acid residueAt P+1 (e.g. X 5 ) The third acidic amino acid residue is located at P+2 (e.g., X 6 ) Hydrophobic amino acid residues are located at P+4 (e.g., X 8 ) The first aromatic amino acid residue is located at P+5 (e.g., X 9 ) The second aromatic amino acid residue is located at P+8 (e.g., X 12 ) And/or a third aromatic amino acid residue is located at P+9 (e.g., X 13 ). In some embodiments, the staple-like structure is an (i, i+7) staple-like structure and another residue of the staple-like structure is located at p+7 (e.g., X 11 ). In some embodiments, the first acidic amino acid residue is at position P-2 (e.g., X 2 ). In some embodiments, the second acidic amino acid residue is located at p+1 (e.g., X 5 ). In some embodiments, the third acidic amino acid residue is located at p+2 (e.g., X 6 ). In some embodiments, the hydrophobic amino acid residue is located at p+4 (e.g., X 8 ). In some embodiments, the first aromatic amino acid residue is located at p+5 (e.g., X 9 ). In some embodiments, the second aromatic amino acid residue is located at p+8 (e.g., X 12 ). In some embodiments, the third aromatic amino acid residue is located at p+9 (e.g., X 13 ). In some embodiments, the first acidic amino acid residue is at position P-2 (e.g., X 2 ) The second acidic amino acid residue is located at P+1 (e.g., X 5 ) The first aromatic amino acid residue is located at P+5 (e.g., X 9 ) The second aromatic amino acid residue is located at P+8 (e.g., X 12 ) And a third aromatic amino acid residue is located at p+9 (e.g., X 13 ). In some embodiments, the first acidic amino acid residue is at position P-2 (e.g., X 2 ) The second acidic amino acid residue is located at P+1 (e.g., X 5 ) The third acidic amino acid residue is located at P+2 (e.g., X 6 ) Hydrophobic amino acid residues are located at P+4 (e.g., X 8 ) The first aromatic amino acid residue is located at P+5 (e.g., X 9 ) The second aromatic amino acid residue is located at P+8 (e.g., X 12 ) And a third aromatic amino acid residue is located at p+9 (e.g., X 13 ). In some embodiments, the stapled peptide agent comprises acidic amino acid residues at positions P-2 and p+1, and aromatic amino acid residues at positions p+5, p+8, and p+9. In some embodiments, the stapled peptide agent comprises acidic amino acid residues at positions P-2, p+1 and p+2, and aromatic amino acid residues at positions p+5, p+8 and p+9. In some embodiments, the stapled peptide agent comprises an acidic amino acid residue at positions P-2 and p+1, a hydrophobic amino acid residue at position p+4, and an aromatic amino acid residue at positions p+5, p+8, and p+9. In some embodiments, the stapled peptide agent comprises an acidic amino acid residue at positions P-2, p+1 and p+2, a hydrophobic amino acid residue at position p+4, and an aromatic amino acid residue at positions p+5, p+8 and p+9. In some embodiments, P is 3. In some embodiments, P is 4. In some embodiments, P is 5. In some embodiments, P is 6. In some embodiments, P is 7. In some embodiments, the amino acid residue at position P comprises two groups for stapling, e.g., B4, B5, B6, etc. In some embodiments, it is B4. In some embodiments, it is B5. In some embodiments, it is B6. In some embodiments, the medicament comprises a staple-like structure and a first additional staple-like structure, e.g., (i, i+3) or (i, i+4) staple-like structure. In some embodiments, the staple-like structure and the first additional staple-like structure are bonded to the same residue (e.g., B5, B6, etc.). In some embodiments, another residue of the first additional residue is at P-2 (e.g., when the moiety used for stapling such as a terminal olefin is at what is considered X 1 In the P terminal group of a portion of (C) a) the P-3 or P-4 position. In some embodiments, the medicament comprises a second additional staple-like structure, such as an (i, i+4) staple-like structure (e.g., at the p+6 position (e.g., X 10 ) And P+10 bits (e.g., X 14 ) Residue stapling of (i, i+3) a staple-like structure (e.g., at position p+3 (e.g., X) 7 ) And the P+6 position (e.g., X 10 ) Residue stapling of (i, i+7) a staple-like structure (e.g., at position p+3 (e.g., X) 7 ) And P+10 bits (e.g., X 14 ) Residues of (2)Stapling), and the like. In some embodiments, the medicament comprises a second additional staple-like structure that is in the p+6 position (e.g., X 10 ) And P+10 bits (e.g., X 14 ) The residue of (i, i+4) stapled, staple-like structure. In some embodiments, the medicament comprises a third additional staple-like structure, e.g., in the P-1 position (e.g., X 3 ) And the P+3 position (e.g., X 7 ) The residue of (i, i+4) stapled, staple-like structure. In some embodiments, there are three staple-like structures in the stapled peptide agent. In some embodiments, there are four staple-like structures in the stapled peptide agent. As demonstrated herein, a stapled medicament comprising such positioned staple-like structures and residues can provide a variety of desirable properties and activities. In some embodiments, the positioning of one or more staple-like structures can be shifted relative to the various acidic, hydrophobic, and/or aromatic amino acid residues described herein, e.g., in some embodiments, the stapled peptide agent comprises a stapling residue at positions P and p+7 (and optionally P-3 or P-4), an acidic amino acid residue at positions P-1 and p+2, and an aromatic amino acid residue at positions p+6, p+9, and p+10, and optionally an acidic amino acid residue at position p+3, and/or a hydrophobic amino acid residue at position p+5. When assessed by fluorescence polarization, it was observed that a variety of stapled peptide agents with shifted staple-like structures could bind to β -catenin.
Some useful staple-like structures are described in the "medicament" section below.
Beta-catenin
The present disclosure provides, inter alia, techniques for modulating one or more β -catenin functions. In some embodiments, the present disclosure provides useful techniques for inhibiting one or more β -catenin functions associated with cancer or hyperplasia. In some embodiments, the provided techniques may be used for the prevention and treatment of conditions, disorders, or diseases for which prevention and/or treatment would benefit from inhibition of β -catenin. In some embodiments, the condition, disorder or disease is cancer.
Beta-catenin has been reported to have multiple functions. For example, it can regulate and coordinate transcription of a variety of genes. High β -catenin activity and/or expression levels have been reported to lead to the occurrence of a variety of conditions, disorders or diseases, including cancer. Mutations and overexpression of β -catenin are reported to be associated with conditions, disorders or diseases, including many cancers, including colorectal, lung and breast cancers. Deregulation of Wnt/β -catenin signalling pathways has been reported to be associated with a number of conditions, disorders or diseases, including neurodegenerative diseases, psychiatric diseases, cancer, asthma and even wound healing. Both a large published clinical and preclinical study have shown that superactivated Wnt/β -catenin activity drives tumorigenesis and is required for tumor maintenance in a variety of cancers. Many Wnt inhibitors greatly modulate this pathway at the extracellular ligand/receptor level, for example, by preventing Wnt ligand secretion or by blocking Wnt ligand interaction with its receptor on the plasma membrane. A number of activated Wnt pathway mutations are reported to exist in APC and/or CTNNB1, downstream of membrane proximal events. The present disclosure specifically covers such recognition: many agents at the extracellular ligand/receptor level are insufficient to treat many relevant patients, e.g., those with downstream mutations/abnormalities. In some embodiments, wnt pathway activating mutations converge on the β -catenin/TCF junction. In some embodiments, the disclosure targets β -catenin/TCF interactions, e.g., as a method of treatment. Agents that modulate β -catenin function may be used for a variety of purposes, including the prevention and/or treatment of a variety of conditions, disorders, or diseases associated with β -catenin.
Binding sites
Beta-catenin can interact with a plurality of agents at a plurality of binding sites, each binding site independently comprising a group of amino acid residues that interact with the binding agent. For example, certain binding sites can be used for interaction of β -catenin with Axin, APC, C-cadherin, E-cadherin, TCF3 and Bcl 9. For interaction with TCF3, it is reported that two or more binding sites can be used simultaneously for interaction with different parts of TCF 3. See, e.g., graham et al cell, vol.103,885-896,2000.
In some embodiments, the provided agents bind to β -catenin at a unique binding site. In some embodiments, the provided agents interact with β -catenin at a different set of amino acid residues than the previously reported binding sites (e.g., those for Axin, APC, C-cadherin, E-cadherin, TCF3, or Bcl 9).
For example, in some embodiments, the agents provided are administered with one or more or all of the groups of amino acid residues (e.g., about 1 to 23, 1 to 20, 1 to 15, 1 to 10, 1 to 5, 5 to 23, 5 to 20, 5 to 15, 5 to 10, 6 to 23, 6 to 20, 6 to 15, 6 to 10, 7 to 23, 7 to 20, 7 to 15, 7 to 10, 8 to 23, 8 to 20, 8 to 15, 8 to 10, 9 to 23, 9 to 20, 9 to 15, 9 to 10, 10 to 23, 10 to 20, 10 to 15, 11 to 23, 11 to 20, 11 to 15, 12 to 23, 12 to 20, 12 to 15, 13 to 23, 13 to 20, 13 to 15, 13 to 23, 14 to 20, 15 to 23, 15 to 20, 16 to 23, 16 to 20, 17 to 23, 17 to 20, 18 to 23, or 18 to 20, or about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 21, 22, 21, or 23) is an amino acid residue of SEQ ID NO:1 or an amino acid residue corresponding to that in SEQ ID No. 1, e.g., in some embodiments, the following amino acid residues of SEQ ID No. 1: a305, Y306, G307, N308, Q309, K312, R342, K345, V346, V349, Q375, R376, Q379, N380, L382, W383, R386, N387, D413, N415, V416, T418, and C419. In some embodiments, the group of amino acid residues is or corresponds to the following: amino acid residues a305, Y306, G307, N308, Q309, K312, R342, K345, V346, V349, Q375, Q379, N380, L382, W383, R386, N387, D413, N415, V416, T418 and C419 of SEQ ID No. 1. In some embodiments, the group of amino acid residues is or corresponds to the following: amino acid residues a305, Y306, G307, N308, Q309, K312, K345, V346, V349, Q379, N380, L382, W383, R386, N387, D413, N415, V416, T418 and C419 of SEQ ID No. 1. In some embodiments, the group of amino acid residues is or corresponds to the following: amino acid residues G307, K312, K345, W383, N387, D413 and N415 of SEQ ID NO. 1. In some embodiments, the group of amino acid residues is or corresponds to the following: amino acid residues G307, K312, K345, Q379, L382, W383, N387, N415 and V416 of SEQ ID NO. 1. In some embodiments, the group of amino acid residues is or corresponds to the following: amino acid residues Y306, G307, K312, K345, Q379, L382, W383, N387, N415 and V416 of SEQ ID NO. 1. In some embodiments, the group of amino acid residues is or corresponds to the following: amino acid residues G307, K312, K345, Q379, L382, W383, R386, N387, N415 and V416 of SEQ ID NO. 1. In some embodiments, the group of amino acid residues is or corresponds to the following: amino acid residues Y306, G307, K312, K345, Q379, L382, W383, R386, N387, N415 and V416 of SEQ ID NO. 1. In some embodiments, the group of amino acid residues is or corresponds to the following: amino acid residues Y306, G307, K312, K345, V349, Q379, L382, W383, N387, N415 and V416 of SEQ ID NO. 1. In some embodiments, the group of amino acid residues is or corresponds to the following: amino acid residues Y306, G307, K312, K345, V349, Q379, L382, W383, R386, N387, N415 and V416 of SEQ ID NO. 1. In some embodiments, the group of amino acid residues is or corresponds to the following: amino acid residues G307, K312, K345, W383 and N387 of SEQ ID NO. 1. In some embodiments, the group of amino acid residues is or corresponds to the following: amino acid residues Y306, G307, K312, R386 and N387 of SEQ ID NO. 1. In some embodiments, the provided agents interact with Y306 or its corresponding amino acid residue. In some embodiments, the provided agents interact with G307 or its corresponding amino acid residue. In some embodiments, the provided agents interact with K312 or its corresponding amino acid residue. In some embodiments, the provided agents interact with K345 or its corresponding amino acid residue. In some embodiments, the provided agents interact with V349 or its corresponding amino acid residue. In some embodiments, the provided agents interact with Q379 or its corresponding amino acid residue. In some embodiments, the provided agents interact with L382 or a corresponding amino acid residue thereof. In some embodiments, the provided agents interact with W383 or its corresponding amino acid residue. In some embodiments, the provided agents interact with R386 or its corresponding amino acid residue. In some embodiments, the provided agents interact with N387 or a corresponding amino acid residue thereof. In some embodiments, the provided agents interact with N415 or its corresponding amino acid residue. In some embodiments, the provided agents interact with V416 or its corresponding amino acid residue.
In some embodiments, the agents of the invention interact with a polypeptide whose sequence corresponds to aa146 to aa665 of human β -catenin. In some embodiments, the agents of the invention interact with a polypeptide whose sequence comprises SEQ ID NO. 2 or SEQ ID NO. 2:
in some embodiments, all amino acid residues that interact with the provided agents have SEQ ID NO. 2. In some embodiments, the amino acid residues that interact with a provided agent (e.g., one or more amino acid residues in the agent) interact with the agent through hydrogen bonds, hydrophobic interactions, or salt bridges. As will be appreciated by those skilled in the art, when two amino acid residues interact with each other, they are typically within a certain distance range when assessed, e.g., using crystallography, NMR, or the like.
In some embodiments, certain amino acid residues that are reported to interact with one or more polypeptides do not significantly participate in the provided interactions with β -catenin. In some embodiments, the provided agent does not interact with an Axin binding site. In some embodiments, the provided agent does not interact with a Bcl9 binding site. In some embodiments, the provided agent does not interact with one or more or all of the following or amino acid residues corresponding to: n426, C429, K435, R469, H470, S473, R474, K508 and N516 of SEQ ID NO. 1. In some embodiments, the provided agent does not interact with N426 or its corresponding amino acid residue. In some embodiments, the provided agents do not interact with C429 or its corresponding amino acid residue. In some embodiments, the provided agent does not interact with K435 or its corresponding amino acid residue. In some embodiments, the provided agents do not interact with R469 or its corresponding amino acid residue. In some embodiments, the provided agent does not interact with H470 or its corresponding amino acid residue. In some embodiments, the provided agent does not interact with S473 or its corresponding amino acid residue. In some embodiments, the provided agent does not interact with R474 or its corresponding amino acid residue. In some embodiments, the provided agent does not interact with K508 or its corresponding amino acid residue. In some embodiments, the provided agent does not interact with N516 or its corresponding amino acid residue.
In some embodiments, mutation of one or more amino acid residues other than SEQ ID NO. 2 in the β -catenin does not significantly (e.g., does not exceed 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90% or more) reduce the interaction of the β -catenin with the provided agent. In some embodiments, it is the following or mutations corresponding to one or more or all of the following amino acid residues that do not significantly reduce the interaction of β -catenin with the provided agent: n426, C429, K435, R469, H470, S473, R474, K508 and N516 of SEQ ID NO. 1. In some embodiments, mutation of N426 or its corresponding amino acid residue does not significantly reduce the interaction of β -catenin with the agent. In some embodiments, mutations in Q379 or its corresponding amino acid residue (e.g., corresponding to Ala, glu, phe, trp, etc.) do not significantly reduce the interaction of β -catenin with an agent.
In some embodiments, the agent binds to the TCF site of β -catenin. In some embodiments, the agent interacts with one or more, but not all, amino acid residues that interact with TCF. In some embodiments, the agent interacts with one or more, but not all, amino acid residues that interact with the extension region of XTcf 3-CBD. In some embodiments, the agent does not interact with β -catenin amino acid residues that interact with β -hairpin modules of XTcf 3-CBD. In some embodiments, the agent does not interact with β -catenin amino acid residues that interact with the helical module of XTcf 3-CBD. For certain amino acid residues that interact with various modules of XTcF3-CBD, see, e.g., graham et al cell, vol.103,885-896,2000.
In some embodiments, the agent competes with TCF for β -catenin binding. In some embodiments, the agent competes for β -catenin binding with an extension region of TCF (e.g., ala14-Glu24 or Asp16-Glu24, as described in Graham et al cell, vol.103,885-896,2000). In some embodiments, the agent does not compete with Axin for β -catenin binding or competes with Axin for β -catenin binding to a lesser extent as compared to the extension region of TCF. In some embodiments, the agent does not compete with Bcl9 for β -catenin binding or competes with Bcl9 for β -catenin binding to a lesser extent as compared to the extension region of TCF. In some embodiments, the agent does not compete for β -catenin binding with the β -hairpin module of the XTcf3-CBD or competes for β -catenin binding to a lesser extent with the β -hairpin module of the XTcf3-CBD as compared to the extension region of TCF. In some embodiments, the agent does not compete for β -catenin binding with or competes for β -catenin binding to a lesser extent with the helical module of XTcf3-CBD compared to the extension region of TCF. In some embodiments, the agent competes for β -catenin binding with E-cadherin.
In some embodiments, the present disclosure provides complexes of a peptide (e.g., a polypeptide whose sequence is SEQ ID NO:1 or 2 or comprises SEQ ID NO:1 or 2) and a provided agent. In some embodiments, in such complexes, the polypeptide and provided agent interact with one or more or all of the amino acid residues as described herein, and optionally do not interact with one or more or all of the amino acid residues as described herein.
In some embodiments, the present disclosure provides a complex comprising a provided agent and a β -catenin polypeptide or a portion thereof. In some embodiments, a portion thereof comprises one or more or all of the interacting residues as described herein. In some embodiments, the agent and the β -catenin polypeptide, or a portion thereof, interact with each other at one or more or all of the interacting residues.
Certain medicaments
In some embodiments, the present disclosure provides a pharmaceutical agent having the structure of formula I:
R N -L P1 -L AA1 -L P2 -L AA2 -L P3 -L AA3 -L P4 -L AA4 -L P5 -L AA5 -L P6 -L AA6 -L P7 -R C ，
I
wherein:
R N is a peptide, an amino protecting group or R' -L RN -；
L P1 、L P2 、L P3 、L P4 、L P5 、L P6 And L P7 Each independently is L, wherein L P1 、L P2 、L P3 、L P4 、L P5 、L P6 And L P7 Comprising:
a first R 'group and a second R' group taken together to form-L s -, the-L s -bonding to an atom to which a first R 'group is attached and an atom to which a second R' group is attached; and
a third R 'group and a fourth R' group taken together to form-L s -, the-L s -bonding to an atom to which a third R 'group is attached and an atom to which a fourth R' group is attached;
each L s Independently is-L s1 -L s2 -L s3 -, each L s1 、L s2 And L s3 Independently is L;
L AA1 is an amino acid residue comprising a side chain containing an acidic or polar group;
L AA2 Is an amino acid residue comprising a side chain containing an acidic or polar group;
L AA3 is an amino acid residue；
L AA4 Is an amino acid residue comprising a side chain comprising an optionally substituted aromatic group;
L AA5 is an amino acid residue comprising a side chain comprising an optionally substituted aromatic group;
L AA6 is an amino acid residue comprising a side chain comprising an optionally substituted aromatic group;
R C is a peptide, a carboxyl protecting group, -L RC -R’、-O-L RC -R 'or-N (R') -L RC -R’；
L RN And L RC Each independently is L;
each L is independently a covalent bond or an optionally substituted divalent C 1 -C 25 Aliphatic or heteroaliphatic groups having from 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R')-, -C (O) S-or-C (O) O-;
each-Cy-is independently an optionally substituted divalent 3-to 30-membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms;
each R' is independently-L-R, -C (O) R, -CO 2 R or-SO 2 R；
Each R is independently-H or an optionally substituted group selected from: c (C) 1-30 Aliphatic, C having 1 to 10 heteroatoms 1-30 Heteroaliphatic, C 6-30 Aryl, C 6-30 Arylaliphatic, C having 1 to 10 heteroatoms 6-30 Aryl heteroaliphatics, 5 to 30 membered heteroaryl groups having 1 to 10 heteroatoms and 3 to 30 membered heterocyclyl groups having 1 to 10 heteroatoms, or
The two R groups optionally and independently together form a covalent bond, or:
two or more R groups on the same atom optionally and independently taken together with the atom form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the atom; or alternatively
Two or more R groups on two or more atoms optionally and independently together with their intervening atoms form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the intervening atoms.
In some embodiments, the present disclosure provides a pharmaceutical agent having the structure of formula I:
R N -L P1 -L AA1 -L P2 -L AA2 -L P3 -L AA3 -L P4 -L AA4 -L P5 -L AA5 -L P6 -L AA6 -L P7 -R C ，
I
wherein:
R N is a peptide, an amino protecting group or R' -L RN -；
L P1 、L P2 、L P3 、L P4 、L P5 、L P6 And L P7 Each independently is L, wherein L P1 、L P2 、L P3 、L P4 、L P5 、L P6 And L P7 Comprising:
a first R 'group and a second R' group taken together to form-L s -, the-L s -bonding to an atom to which a first R 'group is attached and an atom to which a second R' group is attached; and
a third R 'group and a fourth R' group taken together to form-L s -, the-L s -bonding to an atom to which a third R 'group is attached and an atom to which a fourth R' group is attached;
Each L s Independently is-L s1 -L s2 -L s3 -, each L s1 、L s2 And L s3 Independently is L;
L AA1 is L AR Wherein the methylene units are-C(R’)(R AS ) -substitution, wherein R AS is-L AS1 -R AA1 Wherein R is AA1 is-CO 2 R or-SO 2 R；
L AA2 Is L AR Wherein the methylene units are represented by-C (R') (R AS ) -substitution, wherein R AS is-L AS2 -R AA2 Wherein R is AA2 is-CO 2 R or-SO 2 R；
L AA3 Is L AR Wherein the methylene units are represented by-C (R') (R AS ) -substitution, wherein R AS is-L AS3 -R AA3 Wherein R is AA3 Is R';
L AA4 is L AR Wherein the methylene units are represented by-C (R') (R AS ) -substitution, wherein R AS is-L AS4 -R AA4 Wherein R is AA4 Is an optionally substituted group selected from 6 to 14 membered aryl or 5 to 14 membered heteroaryl having 1 to 6 heteroatoms;
L AA5 is L AR Wherein the methylene units are represented by-C (R') (R AS ) -substitution, wherein R AS is-L AS5 -R AA5 Wherein R is AA5 Is an optionally substituted group selected from 6 to 14 membered aryl or 5 to 14 membered heteroaryl having 1 to 6 heteroatoms;
L AA6 is L AR Wherein the methylene units are represented by-C (R') (R AS ) -substitution, wherein R AS is-L AS6 -R AA6 Wherein R is AA6 Is an optionally substituted group selected from 6 to 14 membered aryl or 5 to 14 membered heteroaryl having 1 to 6 heteroatoms;
R C is a peptide, a carboxyl protecting group, -L RC -R’、-O-L RC -R 'or-N (R') -L RC -R’；
L RN And L RC Each independently is L;
each L AR Independently an optionally substituted divalent C 1 -C 4 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-C(R’)(R AS )-、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R')-, -C (O) S-or-C (O) O-;
L AS1 、L AS2 、L AS3 、L AS4 、L AS5 and L AS6 Each independently is L AS ；
Each R AS Independently is-L AS -R’；
Each L AS Independently an optionally substituted divalent C 1 -C 10 Aliphatic or heteroaliphatic groups having from 1 to 5 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R')-, -C (O) S-or-C (O) O-;
each L is independently a covalent bond or an optionally substituted divalent C 1 -C 25 Aliphatic or heteroaliphatic groups having from 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R')-, -C (O) S-or-C (O) O-;
each-Cy-is independently an optionally substituted divalent 3-to 30-membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms;
each R' is independently-L-R, -C (O) R, -CO 2 R or-SO 2 R；
Each R is independently-H or an optionally substituted group selected from: c (C) 1-30 Aliphatic, C having 1 to 10 heteroatoms 1-30 Heteroaliphatic, C 6-30 Aryl, C 6-30 Arylaliphatic, C having 1 to 10 heteroatoms 6-30 Aryl groupHeteroaliphatic, 5-to 30-membered heteroaryl having 1 to 10 heteroatoms and 3-to 30-membered heterocyclyl having 1 to 10 heteroatoms, or
The two R groups optionally and independently together form a covalent bond, or:
two or more R groups on the same atom optionally and independently taken together with the atom form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the atom; or alternatively
Two or more R groups on two or more atoms optionally and independently together with their intervening atoms form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the intervening atoms.
In some embodiments, the second R 'group and the third R' group are attached to the same atom. In some embodiments, none of the first R ' group, the second R ' group, and the fourth R ' group are attached to the same atom. In some embodiments, none of the first, second, fourth, fifth, and sixth R' groups are attached to the same atom. In some embodiments, none of the first, second, fourth, fifth, sixth, seventh, and eighth R' groups are attached to the same atom. In some embodiments, the first R 'group, the second R' group, the third R 'group, and the fourth R' group are each independently attached to a different atom. In some embodiments, the first, second, third, fourth, fifth, and sixth R' groups are each independently linked to a different atom. In some embodiments, the first, second, third, fourth, fifth, sixth, seventh, and eighth R' groups are each independently linked to a different atom.
In some embodiments, the compound of formula I is a stapled peptide as described herein.
In some embodiments, each L s Independently a staple like structure as described herein. In some embodiments, L s For example, by bringing together a first R 'group and a second R' groupL of (2) s Having a length of 5 to 20 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) atoms. Unless otherwise indicated, two attachment sites such as L s The length between L, etc. is the shortest covalent linkage from one site to another. For example, -CH 2 -CH 2 -2 atoms (-C-) in length and 3 atoms in length of the 1, 3-phenylene group. In some embodiments, L s For example, L formed by the third R 'group and the fourth R' group taken together s Having a length of 5 to 20 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) atoms. In some embodiments, L s For example L formed by the fifth R 'group and the sixth R' group together s Having a length of 5 to 20 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) atoms. In some embodiments, L s For example L formed by the seventh R 'group and the eighth R' group together s Having a length of 5 to 20 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) atoms.
Those skilled in the art who review this disclosure will appreciate that a staple-like structure (e.g., L s ) Typically longer than a staple-like structure connecting two atoms having a shorter distance, e.g., (i, i+7) staple-like structures typically have a longer length than (i, i+3) or (i, i+4) staple-like structures. In some embodiments, 5 atoms in length. In some embodiments, 6 atoms in length. In some embodiments, 7 atoms in length. In some embodiments, 8 atoms in length. In some embodiments, 9 atoms in length. In some embodiments, 10 atoms in length. In some embodiments, 11 atoms in length. In some embodiments, 12 atoms in length. In some embodiments, 13 atoms in length. In some embodiments, 14 atoms in length. In some embodiments, 15 atoms in length. In some cases In embodiments, 16 atoms in length. In some embodiments, 17 atoms in length. In some embodiments, 18 atoms in length. In some embodiments, 19 atoms in length. In some embodiments, 20 atoms in length.
L P1
In some embodiments, L P1 Is a covalent bond or an optionally substituted divalent C 2 -C 6 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, L P1 Is 2 to 10 atoms in length. In some embodiments, it is 2 atoms. In some embodiments, it is 3 atoms. In some embodiments, it is 4 atoms. In some embodiments, it is 5 atoms. In some embodiments, it is 6 atoms. In some embodiments, it is 7 atoms. In some embodiments, it is 8 atoms. In some embodiments, it is 9 atoms. In some embodiments, it is 10 atoms. In some embodiments, one or more methylene units are independently substituted with-N (R '), -C (R') 2 -C (O) -or-C (O) N (R') -substitution. In some embodiments, the methylene units are replaced with-N (R') -. In some embodiments, the methylene unit is-C (R') 2 -substitution. In some embodiments, the methylene units are replaced with-C (O) -. In some embodiments, the methylene units are replaced by-C (O) N (R') -. In some embodiments, each methylene unit is independently substituted with-N (R '), -C (R') 2 -or-C (O) -substitution. In some embodiments, L P1 Is or comprises an amino acid residue. In some embodiments, L P1 Is or comprises a peptide.
In some embodiments, L P1 Is- [ X ]] p -X 1 -or comprise- [ X ]] p -X 1 -, where p, X and X 1 Each independently as described herein, and X 1 And L is equal to AA1 And (5) bonding. In some embodiments, L P1 is-X 1 -or comprise-X 1 -。
In some embodiments, L P1 comprising-C (R') 2 -a group wherein one of the R ' groups is the first R ' group of the four R ' groups. In some embodiments, such-C (R') 2 The group belongs to an amino acid residue. In some embodiments, such-C (R') 2 The radical belonging to X 1 . In some embodiments, such carbon atom is the alpha carbon of an amino acid residue.
L AA1
In some embodiments, L AA1 Is or comprises an amino acid residue. In some embodiments, L AA1 Is or comprises the following: amino acid residues comprising side chains containing acidic or polar groups. In some embodiments, L AA1 Is an amino acid residue comprising a side chain containing an acidic group.
In some embodiments, L AA1 Is L AR Wherein the methylene units are represented by-C (R') (R AS ) -instead, wherein each variable is independently as described herein. In some embodiments, L AA1 Is an optionally substituted divalent C 1 -C 6 (e.g., C 1 、C 2 、C 3 、C 4 、C 5 Or C 6 ) An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-C(R’)(R AS )-、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') -, C (O) S-, or-C (O) O-, wherein each variable is independently as described herein. In some embodiments, L AA1 Is an optionally substituted divalent C 2 -C 4 Aliphatic groups wherein one or more methylene units of the group are optionally and independently substitutedThe following alternatives: -C (R') 2 -、-C(R’)(R AS )-、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') -, C (O) S-, or-C (O) O-, wherein each variable is independently as described herein. In some embodiments, L AA1 is-N (R ') -C (R') (R) AS ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, L AA1 is-NH-C (R') (R) AS ) -C (O) -, wherein each variable is independently as described herein.
In some embodiments, L AS1 Is L AS As described herein. In some embodiments, R AA1 is-CO 2 R, wherein R is as described herein. In some embodiments, R is H. In some embodiments, L AA1 Are acidic amino acid residues such as those of Asp, glu, etc. In some embodiments, L AA1 Is X 2 As described herein.
L P2
In some embodiments, L P2 Is a covalent bond or an optionally substituted divalent C 2 -C 6 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, L P2 Is 2 to 10 atoms in length. In some embodiments, it is 2 atoms. In some embodiments, it is 3 atoms. In some embodiments, it is 4 atoms. In some embodiments, it is 5 atoms. In some embodiments, it is 6 atoms. In some embodiments, it is 7 atoms. In some embodiments, it is 8 atoms. In some embodiments, it is 9 atoms. In some embodiments, it is 10 atoms. In some embodiments, one or More methylene units are independently represented by-N (R '), -C (R') 2 -C (O) -or-C (O) N (R') -substitution. In some embodiments, the methylene units are replaced with-N (R') -. In some embodiments, the methylene unit is-C (R') 2 -substitution. In some embodiments, the methylene units are replaced with-C (O) -. In some embodiments, the methylene units are replaced by-C (O) N (R') -. In some embodiments, each methylene unit is independently substituted with-N (R '), -C (R') 2 -or-C (O) -substitution. In some embodiments, L P2 Is or comprises an amino acid residue. In some embodiments, L P2 Is or comprises a peptide.
In some embodiments, L P2 Is- [ X ]]pX 4 [X]p' -or comprise- [ X]pX 4 [X]p '-, where p, p', X and X 4 Each independently as described herein. In some embodiments, L P2 Is- [ X ]]pX 3 X 4 [X]p' -or comprise- [ X]pX 3 X 4 [X]p' -, each X, X 3 And X 4 Independently are amino acid residues, and p' are each independently 0 to 10. In some embodiments, L P2 is-X 3 X 4 -or comprise-X 3 X 4 -, wherein each X 3 And X 4 Independently as described herein, and X 4 And L is equal to AA2 And (5) bonding.
In some embodiments, L P2 comprising-C (R') 2 -a group wherein one of the R 'groups is a second R' group and the other is a third R 'group of the four R' groups. In some embodiments, such-C (R') 2 The group belongs to an amino acid residue. In some embodiments, such-C (R') 2 The radical belonging to X 4 . In some embodiments, such carbon atom is the alpha carbon of an amino acid residue. In some embodiments, such carbon atom is X 4 Is a carbon alpha to the carbon number.
In some embodiments, L P2 is-C (R') 2 -alternatively, wherein one of the R 'groups is a second or fifth or seventh R' group. In one placeIn some embodiments, such-C (R') 2 The group belongs to an amino acid residue. In some embodiments, such-C (R') 2 The radical belonging to X 3 . In some embodiments, such carbon atom is the alpha carbon of an amino acid residue. In some embodiments, such carbon atom is X 3 Is a carbon alpha to the carbon number. In some embodiments, it is a second R' group. In some embodiments, it is a fifth R' group. In some embodiments, it is a seventh R' group.
In some embodiments, L P2 is-C (R') 2 -substitution, wherein one of the R 'groups is a first or a third R' group. In some embodiments, such-C (R') 2 The group belongs to an amino acid residue. In some embodiments, such-C (R') 2 The radical belonging to X 4 . In some embodiments, such carbon atom is the alpha carbon of an amino acid residue. In some embodiments, such carbon atom is X 4 Is a carbon alpha to the carbon number. In some embodiments, it is a first R' group. In some embodiments, it is a third R' group.
L AA2
In some embodiments, L AA2 Is or comprises an amino acid residue. In some embodiments, L AA2 Is or comprises the following: amino acid residues comprising side chains containing acidic or polar groups. In some embodiments, L AA2 Is an amino acid residue comprising a side chain containing an acidic group.
In some embodiments, L AA2 Is L AR Wherein the methylene units are represented by-C (R') (R AS ) -instead, wherein each variable is independently as described herein. In some embodiments, L AA2 Is an optionally substituted divalent C 1 -C 6 (e.g., C 1 、C 2 、C 3 、C 4 、C 5 Or C 6 ) An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-C(R’)(R AS )-、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') -, C (O) S-, or-C (O) O-, wherein each variable is independently as described herein. In some embodiments, L AA2 Is an optionally substituted divalent C 2 -C 4 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-C(R’)(R AS )-、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') -, C (O) S-, or-C (O) O-, wherein each variable is independently as described herein. In some embodiments, L AA2 is-N (R ') -C (R') (R) AS ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, L AA2 is-NH-C (R') (R) AS ) -C (O) -, wherein each variable is independently as described herein.
In some embodiments, L AS2 Is L AS As described herein. In some embodiments, R AA2 is-CO 2 R, wherein R is as described herein. In some embodiments, R is H. In some embodiments, L AA2 Are acidic amino acid residues such as those of Asp, glu, etc. In some embodiments, L AA2 Is X 5 As described herein.
L P3
In some embodiments, L P3 Is a covalent bond. In some embodiments, L P3 Is an optionally substituted divalent C 2 -C 6 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, L P3 Is 0 to 10 atoms in length. In some embodiments, L P3 Is 2 to 10 atoms in length. In some embodiments, it is 2 atoms. In some embodiments, it is 3 atoms. In some embodiments, it is 4 atoms. In some embodiments, it is 5 atoms. In some embodiments, it is 6 atoms. In some embodiments, it is 7 atoms. In some embodiments, it is 8 atoms. In some embodiments, it is 9 atoms. In some embodiments, it is 10 atoms. In some embodiments, one or more methylene units are independently substituted with-N (R '), -C (R') 2 -C (O) -or-C (O) N (R') -substitution. In some embodiments, the methylene units are replaced with-N (R') -. In some embodiments, the methylene unit is-C (R') 2 -substitution. In some embodiments, the methylene units are replaced with-C (O) -. In some embodiments, the methylene units are replaced by-C (O) N (R') -. In some embodiments, each methylene unit is independently substituted with-N (R '), -C (R') 2 -or-C (O) -substitution. In some embodiments, L P3 Is or comprises an amino acid residue. In some embodiments, L P3 Is or comprises a peptide. In some embodiments, L P3 Is- [ X ]]pX 6 X 7 [X]p' -or comprise- [ X]pX 6 X 7 [X]p' -, each X, X 6 And X 7 Independently are amino acid residues, and p' are each independently 0 to 10. In some embodiments, L P3 is-X 6 X 7 -or comprise-X 6 X 7 -, wherein each X 6 And X 7 Independently are amino acid residues. In some embodiments, X 7 And L is equal to AA3 And (5) bonding. In some embodiments, L P3 is-C (R') 2 -alternatively, wherein one of the R 'groups is a fifth, sixth, seventh or eighth R' group. In some embodiments, X 7 comprising-C (R') 2 -, wherein one of the R 'groups is a fifth, sixth, seventh or eighth R' group.
L AA3
In some embodiments, L AA3 Is or comprises an amino acid residue. In some embodiments, L AA3 Is or comprises the following: amino acid residues comprising side chains containing acidic or polar groups. In some embodiments, L AA3 Is an amino acid residue comprising a side chain containing an acidic group.
In some embodiments, L AA3 Is L AR Wherein the methylene units are represented by-C (R') (R AS ) -instead, wherein each variable is independently as described herein. In some embodiments, L AA3 Is an optionally substituted divalent C 1 -C 6 (e.g., C 1 、C 2 、C 3 、C 4 、C 5 Or C 6 ) An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-C(R’)(R AS )-、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') -, C (O) S-, or-C (O) O-, wherein each variable is independently as described herein. In some embodiments, L AA3 Is an optionally substituted divalent C 2 -C 4 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-C(R’)(R AS )-、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') -, C (O) S-, or-C (O) O-, wherein each variable is independently as described herein. In some embodiments, L AA3 is-N (R ') -C (R') (R) AS ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, L AA3 is-NH-C (R') (R) AS ) -C (O) -, wherein each variable is independently as described herein.
In some embodiments, L AS3 Is L AS As described herein. At the position ofIn some embodiments, R AA3 is-CO 2 R, wherein R is as described herein. In some embodiments, R is H. In some embodiments, L AA3 Are acidic amino acid residues such as those of Asp, glu, etc. In some embodiments, L AA3 Is X 6 As described herein.
In some embodiments, L AA3 Comprising hydrophobic groups. In some embodiments, L AA3 Is or comprises a hydrophobic amino acid residue. In some embodiments, L AA3 Is X 8 As described herein.
L P4
In some embodiments, L P4 Is a covalent bond or an optionally substituted divalent C 2 -C 6 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, L P4 Is 0 to 10 atoms in length. In some embodiments, L P4 Is 2 to 10 atoms in length. In some embodiments, it is 2 atoms. In some embodiments, it is 3 atoms. In some embodiments, it is 4 atoms. In some embodiments, it is 5 atoms. In some embodiments, it is 6 atoms. In some embodiments, it is 7 atoms. In some embodiments, it is 8 atoms. In some embodiments, it is 9 atoms. In some embodiments, it is 10 atoms. In some embodiments, one or more methylene units are independently substituted with-N (R '), -C (R') 2 -C (O) -or-C (O) N (R') -substitution. In some embodiments, the methylene units are replaced with-N (R') -. In some embodiments, the methylene unit is-C (R') 2 -substitution. In some embodiments, the methylene units are replaced with-C (O) -. In some embodiments, the methylene unitsIs replaced by-C (O) N (R') -. In some embodiments, each methylene unit is independently substituted with-N (R '), -C (R') 2 -or-C (O) -substitution. In some embodiments, L P4 Is or comprises an amino acid residue. In some embodiments, L P4 Is or comprises a peptide.
In some embodiments, L P4 Is- [ X ]]pX 7 X 8 [X]p' -or comprise- [ X]pX 7 X 8 [X]p' -, each X, X 7 And X 8 Independently are amino acid residues, and p' are each independently 0 to 10. In some embodiments, L P4 is-X 7 X 8 -or comprise-X 7 X 8 -, wherein each X 7 And X 8 Independently as described herein, and X 8 And L is equal to AA4 And (5) bonding.
In some embodiments, L P4 is-C (R') 2 -alternatively, wherein one of the R 'groups is a fifth, sixth, seventh or eighth R' group. In some embodiments, such-C (R') 2 The group belongs to an amino acid residue. In some embodiments, such-C (R') 2 The radical belonging to X 7 . In some embodiments, such carbon atom is the alpha carbon of an amino acid residue. In some embodiments, such carbon atom is X 7 Is a carbon alpha to the carbon number. In some embodiments, it is a fifth R' group. In some embodiments, it is a sixth R' group. In some embodiments, it is a seventh R' group. In some embodiments, it is an eighth R' group.
L AA4
In some embodiments, L AA4 Is or comprises an amino acid residue. In some embodiments, L AA4 Is or comprises the following: amino acid residues comprising side chains containing aromatic groups.
In some embodiments, L AA4 Is L AR Wherein the methylene units are represented by-C (R') (R AS ) -instead, wherein each variable is independently as described herein. In some embodimentsWherein L is AA4 Is an optionally substituted divalent C 1 -C 6 (e.g., C 1 、C 2 、C 3 、C 4 、C 5 Or C 6 ) An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-C(R’)(R AS )-、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') -, C (O) S-, or-C (O) O-, wherein each variable is independently as described herein. In some embodiments, L AA4 Is an optionally substituted divalent C 2 -C 4 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-C(R’)(R AS )-、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') -, C (O) S-, or-C (O) O-, wherein each variable is independently as described herein. In some embodiments, L AA4 is-N (R ') -C (R') (R) AS ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, L AA4 is-NH-C (R') (R) AS ) -C (O) -, wherein each variable is independently as described herein.
In some embodiments, L AS4 Is L AS As described herein. In some embodiments, R AA4 Is optionally substituted C 6-14 Aryl groups. In some embodiments, R AA4 Is an optionally substituted phenyl group. In some embodiments, R AA4 Is phenyl. In some embodiments, R AA4 Is an optionally substituted 10-membered C 10 Bicyclic aryl groups. In some embodiments, R AA4 Is an optionally substituted 5 membered monocyclic heteroaryl having 1 to 4 heteroatoms. In some embodiments, R AA4 Is an optionally substituted 6 membered monocyclic heteroaryl having 1 to 4 heteroatoms. In some embodiments, R AA4 Is an optionally substituted 9 membered bicyclic heteroaryl having 1 to 4 heteroatoms. In some embodiments, R AA4 Is an optionally substituted 10 membered bicyclic heteroaryl having 1 to 4 heteroatoms. In some embodiments, heteroaryl groups have no more than one heteroatom. In some embodiments, the heteroaryl group has two or more heteroatoms. In some embodiments, the heteroatom is oxygen. In some embodiments, the heteroatom is nitrogen. In some embodiments, the heteroatom is sulfur. In some embodiments, R AA4 Is optionally substitutedIn some embodiments, R AA4 Is optionally substituted->In some embodiments, R AA4 Is optionally substitutedIn some embodiments, R AA4 Are aromatic amino acid residues as described herein. In some embodiments, R AA4 Is X 9 As described herein.
L P5
In some embodiments, L P5 Is a covalent bond or an optionally substituted divalent C 2 -C 6 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, L P5 Is 2 to 10 atoms in length. In some embodiments, it is 2 atoms. In some embodiments, it is 3 atoms. In some embodiments, it is 4 atoms. In some embodiments, it is 5 atoms. In some embodiments, it is 6 originsAnd (5) a seed. In some embodiments, it is 7 atoms. In some embodiments, it is 8 atoms. In some embodiments, it is 9 atoms. In some embodiments, it is 10 atoms. In some embodiments, one or more methylene units are independently substituted with-N (R '), -C (R') 2 -C (O) -or-C (O) N (R') -substitution. In some embodiments, the methylene units are replaced with-N (R') -. In some embodiments, the methylene unit is-C (R') 2 -substitution. In some embodiments, the methylene units are replaced with-C (O) -. In some embodiments, the methylene units are replaced by-C (O) N (R') -. In some embodiments, each methylene unit is independently substituted with-N (R '), -C (R') 2 -or-C (O) -substitution. In some embodiments, L P5 Is or comprises an amino acid residue. In some embodiments, L P5 Is or comprises a peptide.
In some embodiments, L P5 Is- [ X ]]pX 11 [X]p' -or comprise- [ X]pX 11 [X]p' -, wherein each variable is independently as described herein. In some embodiments, L P5 is-X 10 X 11 -or comprise-X 10 X 11 -, wherein each X 10 And X 11 Independently as described herein, and X 11 And L is equal to AA5 And (5) bonding.
In some embodiments, L P5 comprising-C (R') 2 -a group wherein one of the R 'groups is a fourth R' group. In some embodiments, L P5 comprising-C (R') 2 -a group wherein one of the R 'groups is a second R' group. In some embodiments, such-C (R') 2 The group belongs to an amino acid residue. In some embodiments, such-C (R') 2 The radical belonging to X 11 . In some embodiments, such carbon atom is the alpha carbon of an amino acid residue. In some embodiments, such carbon atom is X 11 Is a carbon alpha to the carbon number.
In some embodiments, L P5 comprising-C (R') 2 -a group wherein one of the R' groups is fifth, sixth, seventh or seventhAn octar' group. In some embodiments, such-C (R') 2 The group belongs to an amino acid residue. In some embodiments, such-C (R') 2 The radical belonging to X 10 . In some embodiments, such carbon atom is the alpha carbon of an amino acid residue. In some embodiments, such carbon atom is X 10 Is a carbon alpha to the carbon number. In some embodiments, it is a fifth R' group. In some embodiments, it is a sixth R' group. In some embodiments, it is a seventh R' group. In some embodiments, it is an eighth R' group.
L AA5
In some embodiments, L AA5 Is or comprises an amino acid residue. In some embodiments, L AA5 Is or comprises the following: amino acid residues comprising side chains containing aromatic groups.
In some embodiments, L AA5 Is L AR Wherein the methylene units are represented by-C (R') (R AS ) -instead, wherein each variable is independently as described herein. In some embodiments, L AA5 Is an optionally substituted divalent C 1 -C 6 (e.g., C 1 、C 2 、C 3 、C 4 、C 5 Or C 6 ) An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-C(R’)(R AS )-、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') -, C (O) S-, or-C (O) O-, wherein each variable is independently as described herein. In some embodiments, L AA5 Is an optionally substituted divalent C 2 -C 4 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-C(R’)(R AS )-、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') -, C (O) S-, or-C (O) O-, wherein each variable is independently as described herein. In some embodiments, L AA5 is-N (R ') -C (R') (R) AS ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, L AA5 is-NH-C (R') (R) AS ) -C (O) -, wherein each variable is independently as described herein.
In some embodiments, L AS5 Is L AS As described herein. In some embodiments, R AA5 Is optionally substituted C 6-14 Aryl groups. In some embodiments, R AA5 Is an optionally substituted phenyl group. In some embodiments, R AA5 Is phenyl. In some embodiments, R AA5 Is an optionally substituted 10-membered C 10 Bicyclic aryl groups. In some embodiments, R AA5 Is an optionally substituted 5 membered monocyclic heteroaryl having 1 to 4 heteroatoms. In some embodiments, R AA5 Is an optionally substituted 6 membered monocyclic heteroaryl having 1 to 4 heteroatoms. In some embodiments, R AA5 Is an optionally substituted 9 membered bicyclic heteroaryl having 1 to 4 heteroatoms. In some embodiments, R AA5 Is an optionally substituted 10 membered bicyclic heteroaryl having 1 to 4 heteroatoms. In some embodiments, heteroaryl groups have no more than one heteroatom. In some embodiments, the heteroaryl group has two or more heteroatoms. In some embodiments, the heteroatom is oxygen. In some embodiments, the heteroatom is nitrogen. In some embodiments, the heteroatom is sulfur. In some embodiments, R AA5 Is optionally substitutedIn some embodiments, R AA5 Is optionally substituted->In some embodiments, R AA5 Is optionally substituted->In some embodiments, L AA5 Are aromatic amino acid residues as described herein. In some embodiments, L AA5 Is X 12 As described herein.
L P6
In some embodiments, L P6 Is a covalent bond. In some embodiments, L P6 Is an optionally substituted divalent C 2 -C 6 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, L P6 Is 0-10 atoms (e.g., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, etc.). In some embodiments, L P6 Is 2 to 10 atoms in length. In some embodiments, it is 2 atoms. In some embodiments, it is 3 atoms. In some embodiments, it is 4 atoms. In some embodiments, it is 5 atoms. In some embodiments, it is 6 atoms. In some embodiments, it is 7 atoms. In some embodiments, it is 8 atoms. In some embodiments, it is 9 atoms. In some embodiments, it is 10 atoms. In some embodiments, one or more methylene units are independently substituted with-N (R '), -C (R') 2 -C (O) -or-C (O) N (R') -substitution. In some embodiments, the methylene units are replaced with-N (R') -. In some embodiments, the methylene unit is-C (R') 2 -substitution. In some embodiments, the methylene units are replaced with-C (O) -. In some embodiments, the methylene units are replaced by-C (O) N (R') -. In some embodiments, each methylene unit is independently substituted with-N (R '), -C (R') 2 -or-C (O) -substitution. In some embodiments, L P6 Is or comprises an amino acid residue. In some casesIn embodiments, L P6 Is or comprises a peptide.
L AA6
In some embodiments, L AA6 Is or comprises an amino acid residue. In some embodiments, L AA6 Is or comprises the following: amino acid residues comprising side chains containing aromatic groups.
In some embodiments, L AA6 Is L AR Wherein the methylene units are represented by-C (R') (R AS ) -instead, wherein each variable is independently as described herein. In some embodiments, L AA6 Is an optionally substituted divalent C 1 -C 6 (e.g., C 1 、C 2 、C 3 、C 4 、C 5 Or C 6 ) An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-C(R’)(R AS )-、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') -, C (O) S-, or-C (O) O-, wherein each variable is independently as described herein. In some embodiments, L AA6 Is an optionally substituted divalent C 2 -C 4 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-C(R’)(R AS )-、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') -, C (O) S-, or-C (O) O-, wherein each variable is independently as described herein. In some embodiments, L AA6 is-N (R ') -C (R') (R) AS ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, L AA6 is-NH-C (R') (R) AS ) -C (O) -, wherein each variable is independently as described herein.
In some embodiments, L AS6 Is L AS As herein describedSaid method. In some embodiments, R AA6 Is optionally substituted C 6-14 Aryl groups. In some embodiments, R AA6 Is an optionally substituted phenyl group. In some embodiments, R AA6 Is phenyl. In some embodiments, R AA6 Is an optionally substituted 10-membered C 10 Bicyclic aryl groups. In some embodiments, R AA6 Is an optionally substituted 5 membered monocyclic heteroaryl having 1 to 4 heteroatoms. In some embodiments, R AA6 Is an optionally substituted 6 membered monocyclic heteroaryl having 1 to 4 heteroatoms. In some embodiments, R AA6 Is an optionally substituted 9 membered bicyclic heteroaryl having 1 to 4 heteroatoms. In some embodiments, R AA6 Is an optionally substituted 10 membered bicyclic heteroaryl having 1 to 4 heteroatoms. In some embodiments, heteroaryl groups have no more than one heteroatom. In some embodiments, the heteroaryl group has two or more heteroatoms. In some embodiments, the heteroatom is oxygen. In some embodiments, the heteroatom is nitrogen. In some embodiments, the heteroatom is sulfur. In some embodiments, R AA6 Is optionally substitutedIn some embodiments, R AA6 Is optionally substituted->In some embodiments, R AA6 Is optionally substitutedIn some embodiments, L AA6 Are aromatic amino acid residues as described herein. In some embodiments, L AA6 Is X 13 As described herein.
L P7
In some embodiments, L P7 Is a covalent bond. In some embodiments, L P7 Is an optionally substituted divalent C 1 -C 25 (e.g., C 1-20 、C 1-15 、C 1-10 、C 1-5 、C 1 、C 2 、C 3 、C 4 、C 5 、C 6 、C 7 、C 8 、C 9 、C 10 、C 11 、C 12 、C 13 、C 14 、C 15 、C 16 、C 17 、C 18 、C 19 Or C 20 ) Aliphatic or heteroaliphatic groups having from 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, L P7 Is an optionally substituted divalent C 1 -C 25 (e.g., C 1-20 、C 1-15 、C 1-10 、C 1-5 、C 1 、C 2 、C 3 、C 4 、C 5 、C 6 、C 7 、C 8 、C 9 、C 10 、C 11 、C 12 、C 13 、C 14 、C 15 、C 16 、C 17 、C 18 、C 19 Or C 20 ) An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, L P7 Is an optionally substituted divalent C 1 -C 20 Aliphatic or heteroaliphatic groups having from 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, L P7 Is an optionally substituted divalent C 1 -C 15 Aliphatic or heteroaliphatic groups having from 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, L P7 Is an optionally substituted divalent C 1 -C 10 Aliphatic or heteroaliphatic groups having from 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-.
In some embodiments, is-X 14 -[X]p' -or containing-X 14 -[X]p '-, where p' is 0 to 10. In some embodiments, X 14 And L is equal to AA6 And (5) bonding. In some embodiments, L P7 comprising-C (R') 2 -a group wherein one of the R 'groups is a sixth or eighth R' group. In some embodiments, such-C (R') 2 The group belongs to an amino acid residue. In some embodiments, such-C (R') 2 The radical belonging to X 14 . In some embodiments, such carbon atom is the alpha carbon of an amino acid residue. In some embodiments, such carbon atom is X 14 Is a carbon alpha to the carbon number. In some embodiments, it is a sixth R' group. In some embodiments, it is an eighth R' group.
L AS
In some embodiments, L AS Is a covalent bond. In some embodiments, L AS Is an optionally substituted divalent C 1 -C 10 (e.g., C 1-5 、C 1 、C 2 、C 3 、C 4 、C 5 、C 6 、C 7 、C 8 、C 9 Or C 10 ) An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, L AS Is an optionally substituted divalent C 1 -C 10 Aliphatic groups wherein one or more methylene units of the group are optionally and independently substituted with-C (R') 2 -Cy-, -O-, -S-, -N (R'), -C (O) -, -S (O) -or-S (O) 2 -substitution. In some embodiments, L AS Is an optionally substituted divalent C 1 -C 10 Aliphatic groups wherein one or more methylene units of the group are optionally and independently replaced by-O-, -S-, or-N (R') -. In some embodiments, L AS Is an optionally substituted divalent C 1 -C 10 An alkylene group. In some embodiments, L AS Is optionally substituted-CH 2 -. In some embodiments, L AS is-CH 2 -. In some embodiments, L AS Is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 atoms in length. In some embodiments, it is 1 atom. In some embodiments, it is 2 atoms. In some embodiments, it is 3 atoms. In some embodiments, it is 4 atoms. In some embodiments, it is 5 atoms. In some embodiments, it is 6 atoms. In some embodiments, it is 7 atoms. In some embodiments, it is 8 atoms. In some embodiments, it is 9 atoms. In some embodiments, it is 10 atoms.
In some embodiments, the agent of formula I is a stapled peptide as described herein. In some embodiments, the agent of formula I is an agent selected from table E2 or a pharmaceutically acceptable salt thereof. In some embodiments, the agent of formula I is an agent selected from table E3 or a pharmaceutically acceptable salt thereof.
The present disclosure provides, inter alia, agents, such as peptides, that can bind to β -catenin. In some embodiments, the agent is or comprises the following: x is X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 Wherein X is 1 ，X 2 ，X 3 ，X 4 ，X 5 ，X 6 ，X 7 ，X 8 ，X 9 ，X 10 ，X 11 ，X 12 ，X 13 And X 14 Each independently is an amino acid residue. In some embodiments, the agent is or comprises the following: [ X ] 0 ] p0 X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 Wherein p0, p15, p16 and p17 are each independently 0 or 1, and X 0 ，X 1 ，X 2 ，X 3 ，X 4 ，X 5 ，X 6 ，X 7 ，X 8 ，X 9 ，X 10 ，X 11 ，X 12 ，X 13 ，X 14 ，X 15 ，X 16 And X 17 Each independently is an amino acid residue.
A variety of amino acid residues, such as those of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, PA, and the like, may be used in accordance with the present disclosure. Certain useful amino acid residues are described in the present disclosure.
In some embodiments, X 2 And X 5 Each independently is an acidic residue as described herein. In some embodiments, X 2 、X 5 And X 6 Each independently is an acidic residue as described herein. In some embodiments, X 9 、X 12 And X 13 Each independently is a polymer comprising a pendant aromatic-containing group Amino acid residues of the chain.
In some embodiments, X 2 Is an acidic residue. In some embodiments, X 2 Comprising a side chain containing-COOH or a derivative thereof. In some embodiments, X 2 Comprising a side chain containing-COOH. In some embodiments, X 2 Is Asp. X is also described in the present disclosure 2 Is a residue of a different amino acid.
In some embodiments, X 5 Is an acidic residue. In some embodiments, X 5 Comprising a side chain containing-COOH or a derivative thereof. In some embodiments, X 5 Comprising a side chain containing-COOH. In some embodiments, X 5 Is Asp. X is also described in the present disclosure 5 Is a residue of a different amino acid.
In some embodiments, X 6 Is an acidic residue. In some embodiments, X 6 Comprising a side chain containing-COOH or a derivative thereof. In some embodiments, X 6 Comprising a side chain containing-COOH. In some embodiments, X 6 Is Asp. X is also described in the present disclosure 6 Is a residue of a different amino acid.
In some embodiments, X 9 Comprising side chains containing aromatic groups. In some embodiments, X 9 Comprising a side chain comprising-R, wherein R is an optionally substituted group selected from the group consisting of: phenyl, 10 membered bicyclic aryl, 5 membered heteroaryl having 1 to 3 heteroatoms and 9 to 10 membered bicyclic heteroaryl having 1 to 5 heteroatoms. In some embodiments, each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In some embodiments, X 9 Is Phe. X is also described in the present disclosure 9 Is a residue of a different amino acid.
In some embodiments, X 12 Comprising side chains containing aromatic groups. In some embodiments, X 12 Comprising a side chain comprising-R, wherein R is an optionally substituted group selected from the group consisting of: phenyl, 10 membered bicyclic aryl, 5 membered heteroaryl having 1 to 3 heteroatoms and 9 to 10 membered bicyclic heteroaryl having 1 to 5 heteroatomsA base. In some embodiments, each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In some embodiments, X 12 Is 3Thi. In some embodiments, X 12 Is 2F3MeF. In some embodiments, X 12 Is Phe. X is also described in the present disclosure 12 Is a residue of a different amino acid.
In some embodiments, X 13 Comprising side chains containing aromatic groups. In some embodiments, X 13 Comprising a side chain comprising-R, wherein R is an optionally substituted group selected from the group consisting of: phenyl, 10 membered bicyclic aryl, 5 membered heteroaryl having 1 to 3 heteroatoms and 9 to 10 membered bicyclic heteroaryl having 1 to 5 heteroatoms. In some embodiments, each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In some embodiments, X 13 Is BtzA. In some embodiments, X 13 34ClF. In some embodiments, X 13 Is 2NapA. X is also described in the present disclosure 13 Is a residue of a different amino acid.
In some embodiments, the peptide is a stapled peptide, as described herein. In some embodiments, the agent is or comprises a peptide, wherein the peptide is a stapled peptide. In some embodiments, the peptide is a suture peptide. In some embodiments, the peptide comprises three or more staple-like structures as described herein. In some embodiments, the peptide comprises three or more staple-like structures within a region of length, e.g., 11 to 15, e.g., 11, 14, etc., amino acid residues described herein. In some embodiments, such peptides provide improved rigidity, activity, delivery, solubility, and/or other desirable properties as compared to reference peptides that are not stapled or contain fewer staple-like structures.
In some embodiments, the present disclosure provides a medicament, e.g., a peptide, comprising X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 Wherein X is 1 、X 2 、X 3 、X 4 、X 5 、X 6 、X 7 、X 8 、X 9 、X 10 、X 11 、X 12 、X 13 And X 14 Each independently is an amino acid residue and comprises two or more amino acid residue pairs, wherein each amino acid residue pair is independently two amino acid residues suitable for stapling or is stapling. In some embodiments, the present disclosure provides a medicament, e.g., a peptide, comprising X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 Wherein X is 1 、X 2 、X 3 、X 4 、X 5 、X 6 、X 7 、X 8 、X 9 、X 10 、X 11 、X 12 、X 13 And X 14 Each independently is an amino acid residue and comprises two or more amino acid residue pairs, wherein each amino acid residue pair is independently three amino acid residues suitable for stapling or is stapled.
In some embodiments, the present disclosure provides an agent, e.g., a peptide, comprising [ X 0 ] p0 X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 Wherein P0, P15, P16 and P17 are each independently 0 or 1, and X 0 、X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、X 7 、X 8 、X 9 、X 10 、X 11 、X 12 、X 13 、X 14 、X 15 、X 16 And X 17 Each independently is an amino acid residue and comprises two or more amino acid residue pairs, wherein each amino acid residue pair is independently two amino acid residues suitable for stapling or is stapling. In one placeIn some embodiments, the present disclosure provides an agent, e.g., a peptide, comprising [ X 0 ] p0 X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 Wherein P0, P15, P16 and P17 are each independently 0 or 1, and X 0 、X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、X 7 、X 8 、X 9 、X 10 、X 11 、X 12 、X 13 、X 14 、X 15 、X 16 And X 17 Each independently is an amino acid residue and comprises three or more amino acid residue pairs, wherein each amino acid residue pair is independently two amino acid residues suitable for stapling or is stapling. In some embodiments, each amino acid residue in such a pair of amino acid residues is independently selected from X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、X 7 、X 8 、X 9 、X 10 、X 11 、X 12 、X 13 And X 14 。
In some embodiments, there are three such pairs of amino acid residues. In some embodiments, there are four such pairs of amino acid residues. In some embodiments, there are four or more such pairs of amino acid residues. In some embodiments, each pair is independently not stapled. In some embodiments, one or more pairs are independently stapled. In some embodiments, two or more pairs are independently stapled. In some embodiments, three or more pairs are independently stapled. In some embodiments, four or more pairs are independently stapled. In some embodiments, the two pairs are independently stapled. In some embodiments, three pairs are independently stapled. In some embodiments, the four pairs are independently stapled.
In some embodiments, a pair is X 1 And X 4 . In some embodiments, a pair is X 4 And X 11 . In some embodiments, a pair is X 1 And X 3 . In some embodiments, a pair is X 4 And X 11 . In some embodiments, a pair is X 10 And X 14 . In some embodiments, a pair is X 7 And X 10 . In some embodiments, a pair is X 7 And X 14 . In some embodiments, a pair is X 3 And X 7 。
In some embodiments, a pair is X 1 And X 14 And is opposite to X 4 And X 11 . In some embodiments, a pair is X 1 And X 14 For X 4 And X 11 And pair is X 10 And X 14 . In some embodiments, a pair is X 1 And X 14 For X 4 And X 11 And pair is X 7 And X 10 . In some embodiments, a pair is X 1 And X 14 For X 4 And X 11 And pair is X 7 And X 14 . In some embodiments, a pair is X 1 And X 14 For X 4 And X 11 For X 3 And X 7 And pair is X 7 And X 14 . In some embodiments, each pair is independently a pair of amino acid residues suitable for stapling. In some embodiments, each pair is independently stapled.
In some embodiments, a pair is X 1 And X 3 For X 4 And X 11 And pair is X 10 And X 14 . In some embodiments, each pair is independently a pair of amino acid residues suitable for stapling. In some embodiments, each pair is independently stapled.
In some embodiments, the present disclosure provides an agent that is or comprises a peptide comprising:
[X 0 ] p0 X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 ，
wherein:
p0, P15, P16 and P17 are each independently 0 or 1;
X 0 、X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、X 7 、X 8 、X 9 、X 10 、X 11 、X 12 、X 13 、X 14 、X 15 、X 16 and X 17 Each independently is an amino acid residue, wherein the agent binds to β -catenin.
In some embodiments, X 2 Comprising side chains containing acidic or polar groups. In some embodiments, X 2 Comprising a side chain containing an acidic group. In some embodiments, X 2 Comprising a side chain containing a polar group. In some embodiments, X 5 Comprising side chains containing acidic or polar groups. In some embodiments, X 5 Comprising a side chain containing an acidic group. In some embodiments, X 5 Comprising a side chain containing a polar group. In some embodiments, X 13 Comprising a side chain containing an optionally substituted aromatic group. In some embodiments, X 1 、X 3 、X 4 、X 7 、X 10 、X 11 And X 14 Each independently an amino acid residue suitable for stapling, or each independently stapling.
In some embodiments, the present disclosure provides an agent that is or comprises a peptide comprising:
[X 0 ] p0 X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ]p 17 ，
wherein:
p0, p15, p16 and p17 are each independently 0 or 1;
X 0 、X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、X 7 、X 8 、X 9 、X 10 、X 11 、X 12 、X 13 、X 14 、X 15 、X 16 And X 17 Each independently is an amino acid residue, wherein:
X 2 comprising a side chain containing an acidic or polar group;
X 5 comprising a side chain containing an acidic or polar group;
X 13 comprising a side chain containing an optionally substituted aromatic group; and is also provided with
X 1 、X 3 、X 4 、X 7 、X 10 、X 11 And X 14 Each independently is an amino acid residue suitable for stapling, or each independently is stapling. In some embodiments, X 1 、X 3 、X 4 、X 7 、X 10 、X 11 And X 14 Each independently an amino acid residue suitable for stapling, or each independently stapling. In some embodiments, X 1 、X 3 、X 4 、X 7 、X 10 、X 11 And X 14 Each independently is an amino acid residue suitable for stapling, or each independently is stapling. In some embodiments, X 1 、X 3 、X 4 、X 7 、X 10 、X 11 And X 14 Each independently is an amino acid residue suitable for stapling, or each independently is stapling. In some embodiments, X 1 And X 4 Each independently is an amino acid residue suitable for stapling. In some embodiments, X 1 And X 3 Each independently is an amino acid residue suitable for stapling. In some embodiments, X 4 And X 11 Each independently is an amino acid suitable for stapling. In some embodiments, X 1 、X 4 And X 11 Each independently is an amino acid residue suitable for stapling. In some embodiments, X 10 And X 14 Each independently is an amino acid residue suitable for stapling. In some embodiments, X 7 And X 10 Each independently is an amino acid residue suitable for stapling. In some embodiments, X 7 And X 14 Each independently is an amino acid residue suitable for stapling. In some embodiments, X 3 And X 7 Each independently is an amino acid residue suitable for stapling. In some embodiments, X 1 And X 4 Connected by a staple-like structure. In some embodiments, X 1 And X 3 Connected by a staple-like structure. In some embodiments, X 4 And X 11 Connected by a staple-like structure. In some embodiments, X 1 And X 4 Connected by staple-like structure, and X 4 And X 11 Connected by a staple-like structure. In some embodiments, X 10 And X 14 Connected by a staple-like structure. In some embodiments, X 7 And X 10 Connected by a staple-like structure. In some embodiments, X 7 And X 14 Connected by a staple-like structure. In some embodiments, X 3 And X 7 Connected by a staple-like structure.
In some embodiments, the present disclosure provides an agent that is or comprises a peptide comprising:
[X 0 ] p0 X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 ，
wherein:
p0, p15, p16 and p17 are each independently 0 or 1;
X 0 、X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、X 7 、X 8 、X 9 、X 10 、X 11 、X 12 、X 13 、X 14 、X 15 、X 16 and X 17 Each independently is an amino acid residue, wherein:
X 2 comprising a side chain containing an acidic or polar group;
X 5 comprising a side chain containing an acidic or polar group;
X 13 comprising a side chain containing an optionally substituted aromatic group; and wherein:
X 1 and X 4 Connected by staple-like structures, and/or X 4 And X 11 Connected by staple-like structure, and X 10 And X 14 Connected by a staple-like structure.
In some embodiments, the present disclosure provides an agent that is or comprises a peptide comprising:
[X 0 ] p0 X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 ，
wherein:
p0, p15, p16 and p17 are each independently 0 or 1;
X 0 、X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、X 7 、X 8 、X 9 、X 10 、X 11 、X 12 、X 13 、X 14 、X 15 、X 16 and X 17 Each independently is an amino acid residue, wherein:
X 2 comprising a side chain containing an acidic or polar group;
X 5 comprising a side chain containing an acidic or polar group;
X 13 comprising a side chain containing an optionally substituted aromatic group; and wherein:
X 1 and X 4 Connected by staple-like structures, and/or X 4 And X 11 Connected by staple-like structure, and X 7 And X 10 Connected by a staple-like structure.
In some embodiments, the present disclosure provides an agent that is or comprises a peptide comprising:
[X 0 ] p0 X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 ，
wherein:
p0, p15, p16 and p17 are each independently 0 or 1;
X 0 、X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、X 7 、X 8 、X 9 、X 10 、X 11 、X 12 、X 13 、X 14 、X 15 、X 16 and X 17 Each independently is an amino acid residue, wherein:
X 2 comprising a side chain containing an acidic or polar group;
X 5 comprising a side chain containing an acidic or polar group;
X 13 comprising a side chain containing an optionally substituted aromatic group; and wherein:
X 1 and X 4 By staple-like structureConnection, and/or X 4 And X 11 Connected by staple-like structure, and X 7 And X 14 Connected by a staple-like structure.
In some embodiments, the present disclosure provides an agent that is or comprises a peptide comprising:
[X 0 ] p0 X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 ，
wherein:
p0, p15, p16 and p17 are each independently 0 or 1;
X 0 、X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、X 7 、X 8 、X 9 、X 10 、X 11 、X 12 、X 13 、X 14 、X 15 、X 16 and X 17 Each independently is an amino acid residue, wherein:
X 2 comprising a side chain containing an acidic or polar group;
X 5 comprising a side chain containing an acidic or polar group;
X 13 comprising a side chain containing an optionally substituted aromatic group; and wherein:
X 1 and X 4 Connected by staple-like structures, and/or X 4 And X 11 Connected by a staple-like structure; and X is 10 And X 14 Connected by staple-like structures, and/or X 3 And X 7 Connected by a staple-like structure.
In some embodiments, the present disclosure provides an agent that is or comprises a peptide comprising:
[X 0 ] p0 X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 ，
wherein:
p0, p15, p16 and p17 are each independently 0 or 1;
X 0 、X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、X 7 、X 8 、X 9 、X 10 、X 11 、X 12 、X 13 、X 14 、X 15 、X 16 and X 17 Each independently is an amino acid residue, wherein:
X 2 comprising a side chain containing an acidic or polar group;
X 5 comprising a side chain containing an acidic or polar group;
X 13 comprising a side chain containing an optionally substituted aromatic group; and wherein:
X 1 and X 3 Connected by staple-like structure, X 4 And X 11 Connected by a staple-like structure; and X is 10 And X 14 Connected by a staple-like structure.
In some embodiments, X 2 Comprising side chains containing acidic (e.g., -COOH) or polar groups. In some embodiments, X 2 Comprising a side chain containing an acidic group. In some embodiments, X 5 Comprising side chains containing acidic or polar groups. In some embodiments, X 5 Comprising a side chain containing an acidic group. In some embodiments, X 6 Comprising side chains containing acidic or polar groups. In some embodiments, X 6 Comprising a side chain containing an acidic group. In some embodiments, X 9 Comprising a side chain containing an optionally substituted aromatic group. In some embodiments, X 12 Comprising a side chain containing an optionally substituted aromatic group. In some embodiments, X 13 Comprising side chains containing optionally substituted aromatic groups. In some embodiments, X 2 And X 5 Each independently comprising a side chain comprising an acidic or polar group. In some embodiments, X 2 And X 6 Each independently comprising a side chain comprising an acidic or polar group. In some embodiments, X 5 And X 6 Each independently comprising a side chain comprising an acidic or polar group. In some embodiments, X 2 And X 5 Each independently comprising a side chain comprising an acidic group. In some embodiments, X 2 And X 6 Each independently comprising a side chain comprising an acidic group. In some embodiments, X 5 And X 6 Each independently comprising a side chain comprising an acidic group. In some embodiments, X 2 、X 5 And X 6 Each independently comprising a side chain comprising an acidic or polar group. In some embodiments, X 2 、X 5 And X 6 Each independently comprising a side chain comprising an acidic group. In some embodiments, X 9 And X 12 Each independently comprising a side chain comprising an optionally substituted aromatic group. In some embodiments, X 9 And X 13 Each independently comprising a side chain comprising an optionally substituted aromatic group. In some embodiments, X 9 、X 12 And X 13 Each independently comprising a side chain comprising an optionally substituted aromatic group. In some embodiments, X 2 And X 5 Each independently comprises a side chain comprising an acidic group (e.g., -COOH), and X 9 、X 12 And X 13 Each independently comprising a side chain comprising an optionally substituted aromatic group. In some embodiments, X 2 、X 5 And X 6 Each independently comprises a side chain comprising an acidic group (e.g., -COOH), and X 9 、X 12 And X 13 Each independently comprising a side chain comprising an optionally substituted aromatic group.
As described herein, various types of amino acid residues (e.g., those having the structures of formulas a-I, A-II, a-III, a-IV, a-V, A-VI, etc.) may be used in accordance with the present disclosure.X is described herein 0 、X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、X 7 、X 8 、X 9 、X 10 、X 11 、X 12 、X 13 、X 14 、X 15 、X 16 、X 17 Etc.
In some embodiments, p0 is 0. In some embodiments, p0 is 1. Various types of amino acid residues can be used for X 0 . In some embodiments, X 0 Selected from Gly, sar and NMebAla. In some embodiments, X 0 Is Gly. In some embodiments, X 0 Is Sar. In some embodiments, X 0 Is NMebAla. In some embodiments, X 0 Are present in various peptides (e.g., in some embodiments, p0 is 1). In some embodiments, X 0 Are not present in the various peptides (e.g., in some embodiments, p0 is 0).
In some embodiments, X 0 Is an N-terminal residue. In some embodiments, it is bonded to an N-terminal group.
In some embodiments, X 0 Are amino acid residues suitable for stapling.
In some embodiments, amino acid residues suitable for stapling comprise a double bond, e.g., a terminal double bond in a side chain thereof. In some embodiments, it has a structure of-L a -CH＝CH 2 Side chains of the structure. In some embodiments, it is a residue of an amino acid having the structure of formula A-II or A-III, or a salt thereof. In some embodiments, X 0 is-N (R) a1 )-L a1 -C(-L a -CH＝CH 2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 0 is-N (R) a1 )-C(-L a -CH＝CH 2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 0 Is a residue of PL3 and is stapled.
In some embodimentsIn the case of X 0 Is N (-L) a -CH＝CH 2 )(R a1 )-L a1 -C(-L a -CH＝CH 2 )(R a3 )-L a2 -C (O) -, or-N (-L) a -CH＝CH 2 )-L a1 -C(-L a -CH＝CH 2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 0 is-N (-L) a -CH＝CH 2 )-C(-L a -CH＝CH 2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein.
In some embodiments, X 0 Is S5. In some embodiments, X 0 Is S6.
In some embodiments, X 0 Is stapled. Various types of staple-like structures may be used, as described herein. In some embodiments, X 0 And X is 4 Stapling. In some embodiments, X 4 And X is 11 Stapling. In some embodiments, the stapled peptide comprises X 0 -X 4 -X 11 Stapling. In some embodiments, the stapled peptide comprises another staple-like structure, e.g., X 10 -X 14 。
In some embodiments, X 0 Is X 1 As described herein.
Various types of amino acid residues can be used for X 1 For example, amino acids of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or residues of salts thereof, according to the present disclosure. In some embodiments, X 1 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 1 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 1 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some casesIn embodiments, R a3 is-H.
As shown herein (e.g., for a variety of amino acids and residues thereof), in various embodiments, L a Is L, as described herein. For example, in some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments of the present invention, in some embodiments, one or more methylene units of L are independently represented by-C (O) -, -N (R'), -Cy-or-O-substitution. In some embodiments, the methylene units are replaced with-C (O) -. In some embodiments, the methylene units are replaced with-N (R') -. In some embodiments, the methylene units are replaced with-Cy-. In some embodiments, -Cy-is optionally substituted phenylene. In some embodiments, -Cy-is 1, 2-phenylene. In some embodiments, the methylene units are replaced with-O-. In some embodiments, L is-C (O) - (CH) 2 ) n-. In some embodiments, L is-C (O) - (CH) 2 ) 2 -. In some embodiments, L is-C (O) - (CH) 2 ) 3 -. In some embodiments, L is-C (O) -1, 2-phenylene-O-CH 2 -. As will be appreciated by those of skill in the art, the embodiments described for each group or moiety (e.g., L) apply to all groups (e.g., L) that may be such groups or moieties a 、L s1 、L s2 、L s3 Etc.), wherever such embodiments are described.
In some embodiments, X 1 Is a residue comprising an optionally substituted cyclic amino acid. In some embodiments, X 1 Is part of an optionally substituted ring. In some embodiments, X 1 Is an amino acid as described herein (e.g., formula A-I, A-amino acids of II, A-III, etc.). In some embodiments, R a1 And R is a3 Taken together form an optionally substituted ring, e.g., an optionally substituted 3-to 10-membered ring. In some embodiments, R a1 And R is a3 Taken together with intervening atoms, form an optionally substituted 3-to 10-membered saturated or partially saturated ring having 0 to 5 heteroatoms in addition to intervening atoms. In some embodiments, the formed ring is saturated. In some embodiments, the ring formed is monocyclic. In some embodiments, the formed ring has no heteroatoms other than intervening atoms. In some embodiments, L a1 And L a2 Is a covalent bond. In some embodiments, the ring formed is unsubstituted. In some embodiments, the ring formed is substituted. In some embodiments, the substitution includes a double bond that is adapted to double-decompose with another double bond to form a staple-like structure. In some embodiments, X 1 Is MePro.
In some embodiments, X 1 Are amino acid residues suitable for stapling.
In some embodiments, amino acid residues suitable for stapling comprise a double bond, e.g., a terminal double bond, in their side chain. In some embodiments, it has the structure-L a -CH＝CH 2 Is a side chain of (c). In some embodiments, it is a residue of an amino acid having the structure of formula A-II or A-III, or a salt thereof. In some embodiments, X 1 is-N (R) a1 )-L a1 -C(-L a -CH＝CH 2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 1 is-N (R) a1 )-C(-L a -CH＝CH 2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 1 Is a residue of PL3 and is stapled.
In some embodiments, X 1 Is N (-L) a -CH＝CH 2 )(R a1 )-L a1 -C(-L a -CH＝CH 2 )(R a3 )-L a2 -C (O) -or-N (-L) a -CH＝CH 2 )-L a1 -C(-L a -CH＝CH 2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 1 is-N (-L) a -CH＝CH 2 )-C(-L a -CH＝CH 2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein.
In some embodiments, it is PL3. In some embodiments, it is [4 pentenyl ]]MeProIs a residue of (a). In some embodiments, it is [5 hexenyl ]]MeProIs a residue of (a). In some embodiments, it is [ BzAm2OAllyl ]]MeProIs a residue of (a).
In some embodiments, X 1 Is PL3. In some embodiments, X 1 Is S5. In some embodiments, X 1 Is MePro. In some embodiments, X 1 Is Asp. In some embodiments, X 1 Is S6. In some embodiments, X 1 Is Pro. In some embodiments, X 1 Is Ala. In some embodiments, X 1 Is Ser. In some embodiments, X 1 Is ThioPro. In some embodiments, X 1 Is Gly. In some embodiments, X 1 Is NMebAla. In some embodiments, X 1 Is Asn. In some embodiments, X 1 Is TfeGA. In some embodiments, X 1 Is Glu. In some embodiments, X 1 Is an acidic amino acid residue. In some embodiments, X 1 Are polar amino acid residues. In some embodiments, X 1 Comprising hydrophobic side chains.
In some embodiments, the agentComprising an N-terminal group. In some embodiments, X 1 Bonded to the N-terminal group. In some embodiments, X 1 Comprising an N-terminal group. In some embodiments, the N-terminal group is Ac, 4 pentenyl, 5 hexenyl, bzAm2OAllyl, hex, bua, 2PyzCO, 3Phc3, meOPr, lithocholic acid ester, 2FPhc, phC, meSO2, ts, isobutyryl, isovaleryl, etHNCO, tzPyr, 15 pyrapp, 8IAP, 3PydCO, 2PyBu, 2PymCO, 5PymCO, or 4PymCO. In some embodiments, the N-terminal group is Ac,2PyBu,
1Imidac,2F2PyAc,2IAPAc,124TriPr,6QuiAc,3PyAc,123TriAc,1 Pyrazoles, 4PyPrpc,3PyPrpc,5PymAc,1PydoneAc,124TriAc,3IAPAc, me2NAc,4MePipzPrpC, mePipAc, meImid4SO2,8QuiSO2, mPEG4, mPEG8, mPEG16, mPEG24, NPyroR3, C3a, bua, isobutyryl, cpc, bnc, CF3CO,2PyCypCO,Cbc,CypCO,4THPCO,2PyzCO,3Phc3,MeOPr, lithocholic acid ester, 2FPhc, phC, meSO2, ts, isovaleryl, etOH CO,5 hexenyl, tzPyr,15PyraPy,8IAP,3PydCO,2PymCO,5PymCO,4PymCO, or 4 pentenyl.
In some embodiments, the N-terminal group comprises a moiety for stapling, such as a terminal olefin. In some embodiments, the N-terminal group is Ac. In some embodiments, the N-terminal group is NPyroR3. In some embodiments, the N-terminal group is 5 hexenyl. In some embodiments, the N-terminal group is 4-pentenyl.
In some embodiments, X 1 Is that
Ac-PL3, ac-S5, NPyroR3-Asp, ac-MePro,5 hexenyl-MePro, ac-S6,4 pentenyl-MePro, ac-Pro, ac-Ala, bua-PL3, C3a-PL3, cpc-PL3, cbc-PL3, cypCO-PL3,4THPCO-PL3, isobutyryl-PL 3, ac-Asp, ac-Ser, ts-PL3, 15pyraPy-PL3,2PyBu-PL3,4PymCO-PL3,4 pentenyl-ThioPro, 4PyPrpc-PL3,3IAPAc-PL3,4 MePizPrpC-PL 3, mePipAc-PL3, meid 4SO2-PL3, bzam2OAllyl-MePro, ac-Sar, mebAc-Ala, hex-3, NMzCO-3, phc-3, meOPr-PL3, lithocholic acid ester-PL 3,2FPhc-PL3, phC-PL3, meSO2-PL3, isovaleryl-PL 3, etONCO-PL 3, tzPyr-PL3,8IAP-PL3,3PydCO-PL3,2PymCO-PL3,5PymCO-PL3,1Imidac-PL3,2F2PyAc-PL3,2IAPAc-PL3, 124TriPr-PL3,6QuiAc-PL3,3PyAc-PL3, 123TriAc-PL3,1Pyrazoleac-PL3,3PyPrpc-PL3,5PymAc-PL3,1PydoneAc-PL3, 124TriAc-PL3, me 2-PL3, 8QuiSO2-PL3, mPEG4-PL3, mPEG8-PL3, mPEG16-PL3, mPEG-3, mPGR-PL 3, or NPR-NPR 3.
In some embodiments, X 1 Is Ac-PL3. In some embodiments, X 1 Is Ac-S5. In some embodiments, X 1 Is NPyroR3-Asp. In some embodiments, X 1 Is Ac-MePro. In some embodiments, X 1 Is Ac-S6. In some embodiments, X 1 Is 4 pentenyl-MePro. In some embodiments, X 1 Is Ac-Pro. In some embodiments, X 1 Is Ac-Ala. In some embodiments, X 1 Bua-PL3. In some embodiments, X 1 Is C3a-PL3. In some embodiments, X 1 Is Cpc-PL3. In some embodiments, X 1 Is Cbc-PL3. In some embodiments, X 1 Is CypCO-PL3. In some embodiments, X 1 Is 4THPCO-PL3. In some embodiments, X 1 Is isobutyryl-PL 3. In some embodiments, X 1 Bnc-PL3. In some embodiments, X 1 Is CF3CO-PL3.
In some embodiments, X 1 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-I, tables A-II, tables A-III and tables A-IV.
In some embodiments, X 1 Is stapled (staple-like structure and X) 1 Bonding). In some embodiments, X 1 Is a residue of PL3 and is stapled. In some embodiments, X 1 And X is 4 Stapling. In some embodiments, the linking of the amino acid residue pairs (e.g., X 1 And X 4 ) Has a staple-like structure of L s ,L s1 -L s2 -L s3 -structure, wherein L s1 Is an amino acid residue such as X 1 L of (2) a And L is s3 Is another amino acid residue such as X 4 L of (2) a 。
As described herein, in some embodiments, the staple-like structure is L s . In some embodiments, L s1 Is L of one amino acid residue in the stapled amino acid residue pair a And L is s3 Is L of another amino acid residue in the stapled amino acid residue pair a . In some embodiments, L s is-L a -L s2 -L a -wherein each variable is independently as described herein. L is described herein a Is described herein). In some embodiments, L s1 Is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L s3 Is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L s1 And L s3 Each independently is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L s1 And L s3 Each independently is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L s1 is-CH 2 -. In some embodiments, L s3 Is- (CH) 2 ) 3 -。
In some embodiments, L s2 Is L, as described herein. In some embodiments, L is optionally substituted-ch=ch-. In some embodiments, L is an optionally substituted-CH 2 -CH 2 -. In some embodiments, L is-CH 2 -CH 2 -。
In some embodiments, L s is-CH 2 -CH＝CH-(CH 2 ) 3 -. In some embodiments, L s Is- (CH) 2 ) 6 -. In some embodiments, such staple-like structures connect X 1 And X 4 . In some embodiments, such staple-like structures can be linked to other pairs of stapled amino acid residues.
In some embodiments, the staple-like structure (e.g., L s ) Bonded to two backbone atoms. In some embodiments, it is combined with twoAnd the carbon skeleton atoms are bonded. In some embodiments, it is independently bonded to the alpha carbon atom of the amino acid residue at each end. In some embodiments, it is bonded to a nitrogen backbone atom (e.g., α -amino) and a carbon backbone atom (e.g., α -carbon). In some embodiments, it is bonded to two nitrogen backbone atoms (e.g., each independently an a-amino group in some embodiments).
In some embodiments, X 1 Is [ 4-pentenyl ]]MePro, [ 5-pentenyl ]]MePro or [ BzAm2OAllyl ]]MePro. In some embodiments, X 1 And X is 3 Stapling. In some embodiments, linkage X 1 And X 3 Has a staple-like structure with L as described herein s Structure is as follows.
As described herein, in some embodiments, the staple-like structure is L s . In some embodiments, L s1 L being an amino acid residue a As described herein. In some embodiments, L s1 Is L, as described herein. For example, in some embodiments, the first and second substrates, one or more methylene units of L are independently represented by-C (O) -, -N (R'), -Cy-or-O-substitution. In some embodiments, L is-N (R') -C (O) - (CH) 2 ) n -O-CH 2 -, wherein n is 1 to 10. In some embodiments, L is-C (O) - (CH) 2 ) n -O-CH 2 -, wherein n is 1 to 10. In some embodiments, L is-N (R') -C (O) - (CH) 2 ) 2 -O-CH 2 -. In some embodiments, L is-C (O) - (CH) 2 ) 2 -O-CH 2 -. In some embodiments, L is-N (R') -C (O) - (CH) 2 ) 3 -O-CH 2 -. In some embodiments, L is-C (O) - (CH) 2 ) 3 -O-CH 2 -. In some embodiments, L is-N (R') -C (O) - (1, 2-phenylene) -O-CH 2 -. In some embodiments, L is-C (O) - (1, 2-phenylene) -O-CH 2 -. In some embodiments, one or more methylene units of L are substituted with-C (R') 2 -substitution. In some embodiments, one or more methylene units of L are replaced with-CHR'. In some cases In embodiments, R ' (e.g., -N (R '), -C (R ') 2 -etc.) and amino acid residues (e.g. X 1 ) May be R (e.g. R) a1 、R a2 、R a3 Etc.) and intervening atoms together to form an optionally substituted 3-to 10-membered ring having 0-5 heteroatoms, as described herein. In some embodiments, R ' (e.g., -N (R '), -C (R ') 2 -etc.) and amino acid residues bonded to the staple-like structure (e.g., X 1 ) May be R (e.g. R) a1 、R a2 、R a3 Etc.) and intervening atoms together to form an optionally substituted 3-to 10-membered ring having 0-5 heteroatoms, as described herein. In some embodiments, R ' (e.g., -N (R '), -C (R ') 2 -etc.) and amino acid residues bonded to the staple-like structure (e.g., X 1 ) Is a further group R a1 And intervening atoms together form an optionally substituted 3-to 10-membered ring having 0-5 heteroatoms, as described herein. In some embodiments, R ' (e.g., -N (R '), -C (R ') 2 -etc.) and amino acid residues bonded to the staple-like structure (e.g., X 1 ) Is a further group R a2 And intervening atoms together form an optionally substituted 3-to 10-membered ring having 0-5 heteroatoms, as described herein. In some embodiments, R ' (e.g., -N (R '), -C (R ') 2 -etc.) and amino acid residues bonded to the staple-like structure (e.g., X 1 ) Is a further group R a3 And intervening atoms together form an optionally substituted 3-to 10-membered ring having 0-5 heteroatoms, as described herein. In some embodiments, the loop formed is present at an amino acid residue (e.g., X 1 ) Is a ring of (a).
In some embodiments, L s3 Is L, as described herein. In some embodiments, L s3 L being an amino acid residue a As described herein. In some embodiments, L is an optionally substituted divalent straight or branched chainC 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (O) -, -N (R'), -Cy-, or-O-. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (O) -, -N (R'), -Cy-, or-O-. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is-CH 2 -. In some embodiments, L is-CH 2 -N(R’)-CH 2 -. In some embodiments, R' is Bn. In some embodiments, R' is-C (O) R. In some embodiments, R is phenyl. In some embodiments, R is tert-butyl. In some embodiments, R is cyclohexyl.
In some embodiments, L s2 Is optionally substituted-ch=ch-. In some embodiments, L s2 Is optionally substituted-CH 2 -CH 2 -. In some embodiments, L s2 is-CH 2 -CH 2 -。
As indicated herein, in some embodiments, the staple-like structure is bonded to two carbon backbone atoms. In some embodiments, it is independently bonded to the alpha carbon atom of the amino acid residue at each end. In some embodiments, it is bonded to a nitrogen backbone atom (e.g., α -amino) and a carbon backbone atom (e.g., α -carbon). In some embodiments, it is bonded to two nitrogen backbone atoms (e.g., each independently an a-amino group in some embodiments).
In some embodiments, X 1 Is the first amino acid starting from the N-terminus. In some embodiments, X 1 Amino groups of (2) are tertiary amines. In some embodiments, X 1 Amino groups of (2) are primary or secondary amines. In some embodiments, X 1 Is capped. In some embodiments, the capping group is R' as described herein. In some embodimentsIn a scheme, the capping group is-C (O) R, wherein R is as described herein. In some embodiments, R is optionally substituted C 1-6 Aliphatic series. In some embodiments, R is optionally substituted C 1-6 An alkyl group. In some embodiments, R is methyl.
In some embodiments, X 1 Interact with Val349 of β -catenin or its corresponding amino acid residue.
In some embodiments, X 1 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-IV.
Various types of amino acid residues can be used for X 2 For example, amino acids of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or residues of salts thereof, according to the present disclosure. In some embodiments, X 2 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 2 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 2 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H.
In some embodiments, X 2 Are residues of amino acids comprising acidic or polar groups (e.g., amino acids of the formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or salts thereof). In some embodiments, X 2 Are residues of amino acids whose side chains contain acidic groups (which may be referred to as "acidic amino acid residues" in some embodiments).
In some embodiments, the amino acid residue whose side chain comprises an acidic group comprises-COOH in its side chain. In some embodiments, it is a residue of an amino acid having the structure of formulas a-IV or a salt thereof. In some embodiments, it is a residue of an amino acid having the structure of formula PA, PA-a, PA-b, PA-c, and the like. In some embodiments, R PA is-H, and R PS And R is PC is-OH. In some embodiments, it is-N (R a1 )-L a1 -C(-L a -COOH)(R a3 )-L a2 -C (O) -. In some embodiments, it is-NH-L a1 -C(-L a -COOH)(R a3 )-L a2 -C (O) -. In some embodiments, it is-NH-CH (-L) a -COOH)-C(O)-。
L as described herein a Is L, as described herein. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (O) -, -N (R'), -Cy-, or-O-. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (O) -, -N (R'), -Cy-, or-O-. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is- (CH) 2 ) n-. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -。
In some embodiments, the acidic amino acid residue is Asp. In some embodiments, it is Glu. Other acidic amino acid residues are described herein, and may be used at multiple amino acid residue positions.
In some embodiments, X 2 Is a residue of Asp, glu, aad, sbMeAsp, rbMeAsp, aMeDAsp or OAsp. In some embodiments, X 2 Is Asp, glu or Aad residue. In some embodiments, X 2 Is an Asp residue. At the position ofIn some embodiments, X 2 Is a residue of Glu. In some embodiments, X 2 Is a residue of Aad. In some embodiments, X 2 Is a residue of SbMeAsp. In some embodiments, X 2 Is a residue of RbMeAsp. In some embodiments, X 2 Is a residue of aMeDAsp. In some embodiments, X 2 Is a residue of OAsp.
In some embodiments, X 2 Are residues of amino acids whose side chains contain polar groups (e.g., amino acids of the formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or salts thereof) (which may be referred to as "polar amino acid residues" in some embodiments; in some embodiments, amino acid residues whose side chains are charged at, for example, about pH 7.4 are not included).
In some embodiments, the amino acid residue whose side chain comprises a polar group is-N (R a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -. In some embodiments, the amino acid residue whose side chain comprises a polar group is-N (R a1 )-C(R a2 )(R a3 ) -C (O) -. In some embodiments, the side chain comprises an amide group (e.g., -C (O) N (R') 2 For example-CONH 2 ) Amino acid residues of (a) are present. In some embodiments, R a2 is-L a -C(O)N(R’) 2 Wherein each variable is independently as described herein. In some embodiments, R a2 is-L a -C(O)NH 2 Wherein L is independently as described herein. In some embodiments, L a Is L', as described herein. In some embodiments, R a3 Is H. In some embodiments, such a polar amino acid residue is Asn. In some embodiments, it is MeAsn. In some embodiments, the amino acid residue whose side chain comprises a polar group is an amino acid residue whose side chain comprises-OH. In some embodiments, R a2 is-L a -OH, wherein each variable is independently as described herein. In some embodiments, R a2 is-L a -OH, wherein L is independently as described herein. In some embodiments, L a Is L',as described herein. For example, in some embodiments, such amino acid residues are residues of Hse, ser, aThr or Thr. In some embodiments, it is a residue of Hse, ser or agr. In some embodiments, it is a residue of Hse. In some embodiments, it is a residue of Ser. In some embodiments, it is a residue of aThr. In some embodiments, it is a residue of Thr. Other polar amino acid residues are described herein, and may be used at multiple amino acid residue positions.
For example, in some embodiments, X 2 Is a residue of Asn. In some embodiments, X 2 Is a residue of MeAsn. In some embodiments, X 2 Is a residue of Hse, ser, aThr or Thr. In some embodiments, X 2 Is a residue of Hse, ser or aThr. In some embodiments, X 2 Is a residue of Hse. In some embodiments, X 2 Is a residue of Ser. In some embodiments, X 2 Is a residue of aThr. In some embodiments, X 2 Is the residue of Thr.
In some embodiments, X 2 Is that
Asp, ala, asn, glu, npg, ser, hse, val, S5, S6, acLys, tfeGA, aThr, aad, pro, thr, phe, leu, PL3, gln, isoGlu, meAsn, isoDAsp, rbGlu, sbGlu, aspSH, ile, sbMeAsp, rbMeAsp, aMeDAsp, OAsp,3COOHF,NAsp,3Thi,NGlu,isoDGlu,BztA,Tle,Aib,MePro,Chg,Cha, or DipA.
In some embodiments, X 2 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-IV.
In some embodiments, X 2 Interact with Gly307 of beta-catenin or corresponding amino acid residue. In some embodiments, X 2 Interacting with Lys312 of β -catenin or its corresponding amino acid residue. In some embodiments, X 2 Interact with each of Gly307 and Lys312 of β -catenin or its corresponding amino acid residues.
Various types of amino acid residues can be used for X 3 Example (1)Such as amino acids of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or residues of salts thereof, according to the present disclosure. In some embodiments, X 3 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 3 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 3 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H.
In some embodiments, L a Is L, as described herein. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (O) -, -N (R'), -Cy-, or-O-. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (O) -, -N (R'), -Cy-, or-O-. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is-CH 2 -. In some embodiments, L is-CH 2 -N(R’)-CH 2 -. In some embodimentsIn this case, R' is Bn. In some embodiments, R' is-C (O) R. In some embodiments, R is phenyl. In some embodiments, R is tert-butyl. In some embodiments, R is cyclohexyl.
In some embodiments, X 3 Is a hydrophobic amino acid residue.
In some embodiments, a hydrophobic amino acid residue is an amino acid residue whose side chain is an optionally substituted aliphatic group. In some embodiments, the hydrophobic amino acid residue is C whose side chain is optionally substituted 1-10 Residues of amino acids of alkyl groups. In some embodiments, the hydrophobic amino acid residue is one whose side chain is C 1-10 Residues of amino acids of alkyl groups. In some embodiments, the hydrophobic amino acid residue is C whose side chain is optionally substituted with one or more non-polar and non-charged groups 1-10 Residues of aliphatic amino acids. In some embodiments, the hydrophobic amino acid residue is C whose side chain is optionally substituted with one or more non-polar and non-charged groups 1-10 Residues of amino acids of alkyl groups. In some embodiments, a hydrophobic amino acid residue is a C whose side chain is optionally substituted with one or more hydrophobic substituents 1-10 Residues of aliphatic amino acids. In some embodiments, the hydrophobic amino acid residue is one whose side chain is C 1-10 Residues of aliphatic amino acids. In some embodiments, the hydrophobic amino acid residue is one whose side chain is C 1-10 Residues of amino acids of alkyl groups. A variety of hydrophobic amino acid residues may be used in accordance with the present disclosure.
In some embodiments, hydrophobic amino acid residues, e.g., X 3 Has the formula of-NH 2 -C(R a2 )(R a3 ) -C (O) -or-NH-C (R) a2 ) H-C (O) -structures, wherein each variable is independently as described herein. R, as described herein a2 is-L a -R'. In some embodiments, R' is R, as described herein. In some embodiments, R is optionally substituted selected from C 1-10 Aliphatic, phenyl, 10 membered aryl and 5 to 10 membered hetero with 1 to 5 hetero atomsAryl groups. In some embodiments, each substituent (if any) is independently a non-polar group. In some embodiments, R is optionally substituted C 1-10 Aliphatic series. In some embodiments, R is optionally substituted C 1-10 An alkyl group. In some embodiments, R is C 1-10 Aliphatic series. In some embodiments, R is C 1-10 An alkyl group. For example, in some embodiments, R is methyl. In some embodiments, R is isopropyl. In some embodiments, R is 1-methylpropyl. In some embodiments, R is 2-methylpropyl. In some embodiments, R is optionally substituted aryl. In some embodiments, R is aryl. In some embodiments, R is optionally substituted phenyl. In some embodiments, R is phenyl. In some embodiments, R is an optionally substituted 5-to 6-membered heteroaryl having 1-4 heteroatoms. In some embodiments, R is an optionally substituted 5-to 6-membered heteroaryl having 1 heteroatom. In some embodiments, R is a 5-to 6-membered heteroaryl having 1 to 4 heteroatoms. In some embodiments, R is a 5-to 6-membered heteroaryl having 1 heteroatom. In some embodiments, R is an optionally substituted 9-to 10-membered heteroaryl having 1 to 5 heteroatoms. In some embodiments, R is an optionally substituted 9-to 10-membered heteroaryl having 1 heteroatom. In some embodiments, R is a 9 to 10 membered heteroaryl having 1 to 4 heteroatoms. In some embodiments, R is a 9 to 10 membered heteroaryl having 1 heteroatom. In some embodiments, the heteroatom is nitrogen. In some embodiments, the heteroatom is oxygen. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (O) -, -N (R'), -Cy-, or-O-. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (O) -, -N (R'), -Cy-, or-O-. In some embodiments, the hydrophobic amino acid residue is a residue of Ala, val, ile, leu, met, phe, tyr, trp or the like. Other hydrophobic amino acid residues are described herein, and may be used at multiple amino acid residue positions.
In some embodiments, X 3 Comprising a side chain containing a cycloaliphatic group (e.g., a 4, 5, or 6 membered cycloalkyl group). In some embodiments, X 3 Is a residue of Npg, leu, cha, val, nLeu, ile, cypA, cyLeu, chg, diethA, ala, aib, octG or Cba. In some embodiments, X 3 Is a residue of Npg, leu or Cha. In some embodiments, X 3 Is a residue of Npg. In some embodiments, X 3 Is a residue of Leu. In some embodiments, X 3 Is the residue of Cha. In some embodiments, X 3 Is a residue of Val. In some embodiments, X 3 Is a residue of nLeu. In some embodiments, X 3 Is a residue of Ile. In some embodiments, X 3 Is a residue of CypA. In some embodiments, X 3 Is a residue of CyLeu. In some embodiments, X 3 Is a residue of Chg. In some embodiments, X 3 Is a residue of DiethA. In some embodiments, X 3 Is a residue of Ala. In some embodiments, X 3 Are residues of Aib. In some embodiments, X 3 Is a residue of OctG. In some embodiments, X 3 Is a residue of Cba.
In some embodiments, X 3 Comprises a side chain that is or comprises an optionally substituted aromatic group (which may be referred to as an "aromatic amino acid residue" in some embodiments).
In some embodiments, the aromatic amino acid residue has a side chain that is or comprises an optionally substituted aromatic group. In some embodiments, an aromatic amino acid residue, e.g., X 3 Has the formula of-NH 2 -C(R a2 )(R a3 ) -C (O) -or-NH-C (R) a2 ) H-C (O) -structure, wherein each variable is independently as described herein, and R a2 Comprising optionally substituted aromatic groups.
In some embodiments, the aromatic amino acid residue has a side chain that is or comprises an optionally substituted aromatic group, wherein each substituent of the aromatic group is independently halogen. In some embodiments, it comprises a side chain that is or comprises two optionally substituted aromatic groups. In some embodiments, it comprises a side chain that is or comprises an optionally substituted aromatic group, wherein each substituent of the aromatic group is independently selected from halogen or-OH. In some embodiments, the aromatic group is phenyl. In some embodiments, the aromatic group is an optionally substituted 8-to 10-membered bicyclic aryl or heteroaryl having 0 to 5 heteroatoms. In some embodiments, the aromatic group is an optionally substituted 9-to 10-membered bicyclic aryl or heteroaryl having one heteroatom. In some embodiments, it is the residue of an amino acid of formula A-I or a salt thereof. In some embodiments, the amino acid residue has a formula of-NH-C (R a2 )(R a3 ) -C (O) -or-NH-CH (R) a3 ) -C) O) -structure. R, as described herein a3 is-L a -R', wherein each variable is independently as described herein. In some embodiments, R' is an optionally substituted group selected from the group consisting of: phenyl, 10 membered bicyclic aryl, having 1 to 4 heteroatomsA 5 to 6 membered heteroaryl group of a child and a 9 to 10 membered bicyclic heteroaryl group having 1 to 5 heteroatoms. In some embodiments, each substituent is independently halogen or-OH. In some embodiments, R' is optionally substituted phenyl. In some embodiments, R' is phenyl. In some embodiments, R' is optionally substituted aryl. In some embodiments, R' is aryl. In some embodiments, R' is an optionally substituted 5 membered heteroaryl having 1 to 4 heteroatoms. In some embodiments, R' is an optionally substituted 5 membered heteroaryl having 1 heteroatom. In some embodiments, R' is a 5 to 6 membered heteroaryl having 1 to 4 heteroatoms. In some embodiments, R' is a 5-to 6-membered heteroaryl having 1 heteroatom. In some embodiments, R' is an optionally substituted 9-to 10-membered heteroaryl having 1 to 5 heteroatoms. In some embodiments, R' is an optionally substituted 9-to 10-membered heteroaryl having 1 heteroatom. In some embodiments, R' is a 9 to 10 membered heteroaryl having 1 to 4 heteroatoms. In some embodiments, R' is a 9 to 10 membered heteroaryl having 1 heteroatom. In some embodiments, the heteroatom is nitrogen. In some embodiments, the heteroatom is oxygen. In some embodiments, the heteroatom is sulfur. In some embodiments, L a Is a covalent bond. In some embodiments, L a Is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L a Is- (CH) 2 ) n-. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, L a is-CH (Ph) -. In some embodiments, the aromatic amino acid residue is Phe. In some embodiments, the aromatic amino acid residue is Tyr. In some embodiments, the aromatic amino acid residue is Trp. Other aromatic amino acid residues are described herein, and may be used at multiple amino acid residue positions.
In some embodiments, X 3 Is a residue of Phe. In some embodiments, X 3 Is a residue of Pff. In some embodiments, X 3 Is a residue of TyrA base. In some embodiments, X 3 Is a residue of Trp. In some embodiments, X 3 Is the residue of Phg. In some embodiments, X 3 Is a residue of DipA.
In some embodiments, X 3 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-I, tables A-II, tables A-III and tables A-IV.
In some embodiments, X 3 Are residues of amino acids suitable for stapling. In some embodiments, X 3 Are residues of amino acids comprising a double bond (e.g., a terminal olefin) that are suitable for stapling. In some embodiments, X 3 Is a residue of an amino acid having a structure such as A-II or A-III or a salt thereof. In some embodiments, X 3 is-N (R) a1 )-L a1 -C(-L a -CH＝CH 2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein.
In some embodiments, X 3 is-N (R) a1 )-C(-L a -CH＝CH 2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 3 Is the residue of AllylGlyThe residues being). In some embodiments, X 3 Is [ Bn ]][ allyl group]DapIn some embodiments, X 3 Is [ Phc ]][ allyl group]DapIn some embodiments, X 3 Is [ Piv ]][ allyl group]DapIn some embodiments, X 3 Is [ CyCO ]][ allyl group]Dap
In some embodiments, X 3 Is stapled. In some embodiments, X 3 And X is 1 Stapling (e.g., by olefin metathesis, where X 1 And X 3 Both contain-ch=ch 2 ). In some embodiments, the staple-like structure has-L s1 -L s2 -L s3 -a structure, wherein each variable is as described herein. In some embodiments, L s1 Is a stapled amino acid residue (e.g.X 1 ) L of (2) a And L is s3 Is another stapled amino acid residue (e.g.X 3 ) L of (2) a . For example, in some embodiments, L s is-C (O) - (CH) 2 )n-L s2 -(CH 2 ) n-, wherein each variable is independently as described herein. In some embodiments, L s is-C (O) - (CH) 2 )n-L s2 -CH 2 -N(R’)-CH 2 -wherein each variable is independently as described herein. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, L s is-C (O) -Cy-O-CH 2 -L s2 -CH 2 Each variable is independently as described herein. In some embodiments, L s is-C (O) -Cy-O-CH 2 -L s2 -CH 2 -N(R’)-CH 2 Each variable is independently as described herein. In some embodiments, -Cy-is optionally substituted phenylene. In some embodiments, -Cy-is 1, 2-phenylene. In some embodiments, R' is Bn. In some embodiments, R' is-C (O) R. In some embodiments, R is phenyl. In some embodiments, R is tert-butyl. In some embodiments, R is cyclohexyl. In some embodiments, L s2 Is optionally substituted-ch=ch-. In some embodiments, L s2 Is-ch=ch-.In some embodiments, L s2 Is optionally substituted-CH 2 -CH 2 -. In some embodiments, L s2 is-CH 2 -CH 2 -. In some embodiments, one end of the staple-like structure, e.g., L s Bonded to a backbone nitrogen atom (e.g., of an alpha amino group at- -C (O) - -of a staple-like structure), and the other end is bonded to a backbone carbon atom (e.g., of- -CH of a staple-like structure) 2 -an alpha carbon atom at) bond.
In some embodiments, amino acid residues suitable for stapling, e.g., X 3 Is an amino acid of formula V or VI or a salt thereof. In some embodiments, such amino acid residues are-N (R a1 )-L a1 -C(-L a -R SP1 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, such amino acid residues are-N (R a1 )-C(-L a -R SP1 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, the reactive group R SP1 is-COOH. In some embodiments, the amino acid suitable for stapling is an amino acid of formula IV or a salt thereof. In some embodiments, such an amino acid is GlnR. In some embodiments, such an amino acid residue may be combined with another amino acid residue suitable for stapling (e.g., comprising an amino acid residue that is-NH 2 R of (2) SP1 A group (e.g., in Lys)).
In some embodiments, X 3 Is GlnR.
In some embodiments, X 3 And X is 7 Stapling. In some embodiments, X 3 comprises-COOH which forms a staple-like structure with a side chain of another amino acid, e.g. Lys, comprising an amino group.
As described herein, in some embodiments, the staple-like structure, e.g., L s comprising-C (O) N (R ') -, wherein R' is as described herein. In some embodiments, R' is-H. In some embodiments, staple-like structures, e.g., L s having-L s1 -C(O)N(R’)-L s3 -structure wherein each variable is independently as described herein. In some embodiments, L s1 Is L, as described herein. In some embodiments, L s3 Is L, as described herein. In some embodiments, L s1 L as described herein is one amino acid residue in a stapled pair a . In some embodiments, L s1 L as described herein is the other amino acid residue in the stapled pair a . In some embodiments, L s1 Independently an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L s3 Independently an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L s1 And L s3 Each independently is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L s1 And L s3 Each independently is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L s1 is-CH 2 -. In some embodiments, L s3 Is- (CH) 2 ) 3 -。
In some embodiments, L s2 Is L, as described herein. In some embodiments, L is-C (O) N (R ') -or comprises-C (O) N (R ') -, wherein R ' is as described herein. In some embodiments, L is or comprises-C (O) NH-.
In some embodiments, L s Is- (CH) 2 ) n1 -C(O)NH-(CH 2 ) n2 -wherein n1 and n2 are each independently n, as described herein. In some embodiments, L s Is- (CH) 2 ) 2 -C(O)NH-(CH 2 ) 4 -. In some embodiments, such staple-like structures connect X 3 And X 7 . In some embodiments, such staple-like structures can be linked to other pairs of stapled amino acid residues.
In some embodiments, X 3 Is the residue of an amino acid comprising an acidic or polar group. In some casesIn embodiments, X 3 Is a residue of an amino acid whose side chain contains an acidic group (e.g., a-COOH group) or a salt form thereof (e.g., a compound of formula A-IV, PA-a, PA-b, PA-c, etc.). In some embodiments, X 3 is-N (R) a1 )-L a1 -C(-L a -COOH)(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 3 is-N (R) a1 )-C(-L a -COOH)(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 3 Is an Asp residue. In some embodiments, X 3 Is the residue of an amino acid whose side chain contains-OH. For example, in some embodiments, X 3 Is a residue of Tyr. In some embodiments, X 3 Is a residue of Ser.
In some embodiments, X 3 Is a residue selected from the group consisting of: npg, leu, cha, allylGly, glnR, val, nLeu, asp, [ Bn ]][ allyl group]Dap、[Phc][ allyl group]Dap、Ile、Phe、CypA、CyLeu、Chg、Pff、DiethA、Ala、Tyr、Trp、Ser、Aib、Phg、OctG、Cba、MorphNva、F2PipNva、[Piv][ allyl group]Dap and [ CyCO ]][ allyl group]Dap。
In some embodiments, X 3 Is the residue: npg, ile, asp, cha, dipA, chg, leu, B5, cba, S5, ala, glu, allylGly, nLeu, ser, B, asn, B4, glnR, val, [ Phc ]][ allyl group]Dap、Hse、[Bn][ allyl group]Dap、1MeK、R5、Phe、CypA、CyLeu、Pff、DiethA、Tyr、Trp、Aib、Phg、OctG、MorphNva、F2PipNva、[Piv][ allyl group]Dap、[CyCO][ allyl group]Dap, lys or S3. In some embodiments, X 3 Is Npg. In some embodiments, X 3 Is Leu. In some embodiments, npg provides better properties and/or activity than, for example, ala.
In some embodiments, X 3 Interact with Tyr306 of β -catenin or its corresponding amino acid residue.
In some embodiments, X 3 Is or comprises the following: residues of moieties or amino acids selected from tables A-IV 。
Various types of amino acid residues can be used for X 4 For example, amino acids of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or residues of salts thereof, according to the present disclosure. In some embodiments, X 4 Is the residue of an amino acid of the formula A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or a salt thereof. In some embodiments, X 4 Is the residue of an amino acid of formula A-II or a salt thereof. In some embodiments, X 4 Are residues of amino acids of formulae A-III or salts thereof. In some embodiments, X 4 Are residues of amino acids of formulae A-IV or salts thereof. In some embodiments, X 4 Are residues of amino acids of formulae A-V or salts thereof. In some embodiments, X 4 Are residues of amino acids of formulae A-VI or salts thereof. In some embodiments, X 4 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 4 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 4 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently as described herein. In some embodiments, R a2 is-L a -CH＝CH 2 Wherein L is a As described herein. In some embodiments, R a3 is-L a -CH＝CH 2 Wherein L is a As described herein. In some embodiments, X 4 is-N (R) a1 )-L a1 -C(-L a -R SP1 )(-L a -R SP2 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 4 is-N (R) a1 )-C(-L a -R SP1 )(-L a -R SP2 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H.
In some embodiments, R SP1 And R is SP2 Each of which is a single pieceIndependently is optionally substituted-ch=ch 2 Or independently comprises optionally substituted-ch=ch 2 . In some embodiments, R SP1 And R is SP2 Each independently is-ch=ch 2 . In some embodiments, -L a R of the linkage SP1 Or R is SP2 Each independently is L, as described herein. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (O) -, -N (R'), -Cy-, or-O-. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (O) -, -N (R'), -Cy-, or-O-.
In some embodiments, X 4 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-I, tables A-II, tables A-III and tables A-IV.
In some embodiments, X 4 Are residues of amino acids suitable for stapling. In some embodiments, X 4 Is a residue of an amino acid comprising two functional groups suitable for stapling. In some embodiments, X 4 Is a residue of an amino acid comprising one and only one functional group suitable for stapling. In some embodiments, X 4 Is a catalyst comprising two olefins (e.g.,two terminal olefins). In some embodiments, X 4 Is a residue of an amino acid that contains one and only one double bond (e.g., terminal alkene) for stapling. In some embodiments, X 4 Is a residue of an amino acid having the structure of formula A-I, A-II, A-III, etc., wherein R a2 And R is a3 Both are independently-L a -CH＝CH 2 Wherein each L a Independently as described herein. In some embodiments, X 4 Is a residue of an amino acid having the structure of formula A-I, A-II, A-III, etc., wherein R a2 And R is a3 Only one of which is-L a -CH＝CH 2 Wherein each L a Independently as described herein. In some embodiments, each L a Independently an optionally substituted divalent C 1-10 Alkylene or heteroalkylene. In some embodiments, each L a Independently is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 6. In some embodiments, n is 7. In some embodiments, n is 8. In some embodiments, n is 9. In some embodiments, n is 10. In some embodiments, X 4 Is a residue of B5, R4 or R6. In some embodiments, X 4 Is a residue of B5 or R5. In some embodiments, X 4 Is the residue of B5. In some embodiments, X 4 Is a residue of R5. In some embodiments, X 4 Is a residue of R4. In some embodiments, X 4 Is a residue of R6.
In some embodiments, X 4 Is stapled. In some embodiments, X 4 Independently linked to two residues by two staple-like structures (e.g., when X 4 And B5). In some embodiments, X 4 And X is 1 Stapling, and X 4 And X is 11 Stapling.
As described herein in the context of the present application,a variety of staple-like structures are available for linking the stapled amino acid residues. In some embodiments, the staple-like structure is L s As described herein. In some embodiments, with X 4 Each staple-like structure connected is independently L s As described herein.
In some embodiments, L s is-L s1 -L s2 -L s3 -wherein each variable is independently as described herein. In some embodiments, L s1 And L s3 One of which is L of one of two stapled amino acid residues a And the other is L of the other of the two stapled amino acid residues a . In some embodiments, L s3 Is X 4 L of (2) a For example, when X 4 With amino acid residues on the N-terminal side thereof (e.g.X 1 ) When stapling. In some embodiments, L s1 Is X 4 L of (2) a For example, when X 4 With amino acid residues on the C-terminal side thereof (e.g.X 11 ) When stapling. In some embodiments, L s1 Is X 1 L of (2) a And L is s3 Is X 4 L of (2) a . In some embodiments, L s1 Is X 4 L of (2) a And L is s3 Is X 11 L of (2) a . In some embodiments, two staple-like structures are associated with X 4 Bonding, wherein the first staple-like structure bonds X 4 And X is 4 Is stapled (the amino acid residue on the N-terminal side of the reference amino acid residue may be referred to as the "N-directional amino acid residue" of the reference amino acid residue, e.g., X 1 Is X 4 N-directional amino acid residues of (c), wherein the first staple-like structure is of the formula-L s1 -L s2 -L s3 L of the Structure s Wherein L is s1 L is an N-directional amino acid residue a And L is s3 Is X 4 L of (2) a And wherein the second staple-like structure will be X 4 And X is 4 Is stapled to the amino acid residue on the C-terminal side of (the amino acid residue on the C-terminal side of the reference amino acid residue may be referred to as "C" of the reference amino acid residueDirectional amino acid residues ", e.g. X 11 Is X 4 C-directed amino acid residues) of (C), wherein the second staple-like structure is of-L s1 -L s2 -L s3 L of the Structure s Wherein L is s3 L being a C-direction amino acid residue a And L is s1 Is X 4 L of (2) a . L is described herein a And can be used for multiple amino acid residues, including X 4 In the N direction (e.g. X 1 ) And C direction (e.g. X 11 ) Amino acid residues. For example, in some embodiments, for X 4 Each L a Is- (CH) 2 ) 3 -。
In some embodiments, L as described herein s2 Is optionally substituted-ch=ch-. In some embodiments, L s2 Is-ch=ch-. In some embodiments, L s2 Is optionally substituted-CH 2 -CH 2 -. In some embodiments, L s2 is-CH 2 -CH 2 -。
In some embodiments, each staple-like structure is independently bonded to two alpha carbon atoms of two stapled amino acid residues, as described herein.
In some embodiments, X 4 With two amino acid residues, e.g. X 1 And X 11 Stapling. In some embodiments, X 4 With only one residue, e.g. X 11 Stapling (e.g., when X 4 R5, R4 or R6). In some embodiments, X 4 is-N (R) a1 )-L a1 -C(-L a -CH＝CH 2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 4 is-N (R) a1 )-C(-L a -CH＝CH 2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 4 Is a residue of R4. In some embodiments, X 4 Is a residue of R5. In some embodiments, X 4 Is a residue of R6.
In some embodiments, the staple-like structure is L s As described herein. For example, in some embodiments, L s1 A first amino acid residue which is two stapled amino acid residues (e.g.X 4 ) L of (2) a And L is s3 A second amino acid residue which is two stapled amino acid residues (e.g.X 11 ) L of (2) a Wherein the second amino acid residue (e.g., X 11 ) Is a first amino acid residue (e.g., X 4 ) C-oriented amino acid residues of (C).
In some embodiments, X 4 Not stapled (e.g., in pre-stapling agents, when other residues are optionally stapled, etc.).
In some embodiments, X 4 Is B5, npg, asp, R5, ile, ala, cha, chg, ser, leu, R4, R6, phe, or S5.
Various types of amino acid residues can be used for X 5 For example, amino acids of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or residues of salts thereof, according to the present disclosure. In some embodiments, X 5 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 5 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 5 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H.
In some embodiments, X 5 Is the residue of an amino acid comprising an acidic or polar group. In some embodiments, X 5 Is a residue of an amino acid whose side chain contains an acidic group (e.g., -COOH group) or a salt form thereof. In some embodiments, X 5 Are residues of amino acids of formulae A-IV or salts thereof. In some embodiments, X 5 Is an amino acid of the formula PA, PA-a, PA-b, PA-c orResidues of salts thereof. In some embodiments, R PA is-H, and R PS And R is PC is-OH. In some embodiments, X 5 is-N (R) a1 )-L a1 -C(-L a -COOH)(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 5 is-N (R) a1 )-C(-L a -COOH)(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, L a Is L, as described herein. For example, in some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments, L is-CH (CH 3 ) -. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (O) -, -N (R'), -Cy-, or-O-.
In some embodiments, X 5 Is a residue of Asp, glu, aad, sbMeAsp or RbMeAsp. In some embodiments, X 5 Is Asp or Glu residue. In some embodiments, X 5 Is an Asp residue. In some embodiments, X 5 Is a residue of Glu. In some embodiments, X 5 Is a residue of Aad. In some embodiments, X 5 Is a residue of SbMeAsp. In some embodiments, X 5 Is a residue of RbMeAsp.
In some embodimentsIn the case of X 5 Is the residue of an amino acid whose side chain contains a polar group. In some embodiments, X 5 Is a compound whose side chain contains an amide group (e.g., -C (O) N (R') 2 For example-CONH 2 ) Residues of amino acids of (a). In some embodiments, R a2 is-L a -C(O)N(R’) 2 Wherein each variable is independently as described herein. In some embodiments, R a2 is-L a -C(O)NH 2 Wherein L is independently as described herein. In some embodiments, L a Is L', as described herein. For example, in some embodiments, X 5 Is a residue of Asn. In some embodiments, X 5 Is a residue of MeAsn. In some embodiments, X 5 Is the residue of an amino acid whose side chain contains-OH. For example, in some embodiments, X 5 Is a residue of Hse, aThr, ser or Thr. In some embodiments, X 5 Is a residue of Hse or aThr. In some embodiments, X 5 Is a residue of Hse. In some embodiments, X 5 Is a residue of aThr. In some embodiments, X 5 Is a residue of Ser. In some embodiments, X 5 Is the residue of Thr.
In some embodiments, X 5 Asp, B5, 3COOHF, glu, asn, npg, hse, aThr, aad, ser, thr, meAsn, aspSH, sbMeAsp, rbMeAsp. In some embodiments, X 5 Is Asp. In some embodiments, X 5 Is 3COOHF. In some embodiments, X 5 Is Glu. In some embodiments, X 5 Is B5. In some embodiments, X 5 Is DipA. In some embodiments, X 5 Is Chg.
In some embodiments, X 5 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-IV.
In some embodiments, X 5 Interact with Trp383 of beta-catenin or its corresponding amino acid residue. In some embodiments, X 5 Interact with Arg386 of beta-catenin or its corresponding amino acid residue. In some embodimentsIn the case of X 5 Interact with Asn387 of β -catenin or its corresponding amino acid residue. In some embodiments, X 5 Interact with Asn387 and Trp383 of β -catenin or their corresponding amino acid residues.
Various types of amino acid residues can be used for X 6 For example, amino acids of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or residues of salts thereof, according to the present disclosure. In some embodiments, X 6 Is the residue of an amino acid of the formula A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or a salt thereof. In some embodiments, X 6 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 6 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 6 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently as described herein. In some embodiments, X 6 Are residues of amino acids of formulae A-IV or salts thereof. In some embodiments, X 6 Is a residue of an amino acid of the formula PA, PA-a, PA-b, PA-c or a salt thereof. In some embodiments, R PA is-H, and R PS And R is PC is-OH. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H.
In some embodiments, X 6 Is the residue of an amino acid comprising an acidic or polar group. In some embodiments, X 6 Is a residue of an amino acid whose side chain contains an acidic group (e.g., -COOH group) or a salt form thereof. In some embodiments, X 6 Is a residue of an amino acid having the structure of formula A-IV or a salt thereof. In some embodiments, X 6 Is a residue of an amino acid having the structure of formula PA, PA-a, PA-b, PA-c, etc. In some embodiments, R PA is-H, and R PS And R is PC is-OH. In some embodiments, X 6 is-N (R) a1 )-L a1 -C(-L a -COOH)(R a3 )-L a2 -C (O) -. In some embodiments, X 6 is-NH-L a1 -C(-L a -COOH)(R a3 )-L a2 -C (O) -. In some embodiments, X 6 is-NH-CH (-L) a -COOH)-C(O)-。
L as described herein a Is L, as described herein. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (O) -, -N (R'), -Cy-, or-O-. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (O) -, -N (R'), -Cy-, or-O-. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is- (CH) 2 ) n-. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments, the methylene units are replaced with-Cy-. In some embodiments, L is-CH 2 -Cy-CH 2 -. In some embodiments, L is-CH 2 -Cy-. In some embodiments, L is- (CH) 2 ) 4 -Cy-CH 2 -C(CH 3 ) 2 -. In some embodiments, -Cy-is optionally substituted phenylene. In some embodiments, -Cy-is phenylene. In some embodiments, -Cy-is a substituted phenylene group. In some embodiments, -Cy-is a monosubstituted phenylene. In some embodiments, the substituent is-F. In some embodiments, the substituent is optionally substituted C 1-6 Alkyl group. In some embodiments, the substituent is-CF 3 . In some embodiments, the substituent is —oh. In some embodiments, the phenylene group is a 1, 2-phenylene group. In some embodiments, the phenylene group is a 1, 3-phenylene group. In some embodiments, the phenylene group is a 1, 4-phenylene group. In some embodiments, the substituents are ortho to the carbon atom near-COOH. In some embodiments, it is meta. In some embodiments, it is para. In some embodiments, -Cy-is 1, 3-phenylene (e.g., in 3 COOHF). In some embodiments, -Cy-is an optionally substituted divalent 5-to 10-membered heteroaryl group having 1-5 heteroatoms. In some embodiments, -Cy-is an optionally substituted divalent 5 membered heteroaryl group having 1 to 4 heteroatoms. In some embodiments, -Cy-is an optionally substituted divalent 6 membered heteroaryl group having 1 to 4 heteroatoms. In some embodiments, -Cy-is optionally substitutedIn some embodiments, -Cy-is +.>In some embodiments, -Cy-is optionally substituted +.>In some embodiments, -Cy-is +.>In some embodiments, L is in the form of-CH 2 Bonded to a backbone atom (e.g., an alpha carbon atom). In some embodiments, the methylene units are replaced with-N (R ') -, wherein R' is as described herein. In some embodiments, L is-CH 2 -N(R’)-CH 2 -, wherein R' is as described herein. In some embodiments, R' is R, as described herein. In some embodiments, R is optionally substituted C 1-6 An alkyl group. In some embodiments, R is-CH 2 CF 3 。
In one placeIn some embodiments, X 6 Is the residue of an amino acid of the formula PA, PA-a, PA-b, PA-c or a salt thereof, wherein R PA is-H, and R PS And R is PC is-OH. In some embodiments, X 6 Is the residue:
3COOHF, tfeGA, asp, [ CH2CMe2CO2H ] TriAzDap, glu,2OH3COOHF, 4COOHF,2COOHF,5F3Me2COOHF,4F3Me2COOHF,5F3Me3COOHF,4F3Me3COOHF,3F2COOHF, or dGlu.
In some embodiments, X 6 Is 3COOHF, tfeGA, asp or [ CH2CMe2CO2H]Residues of TriAzDap. In some embodiments, X 6 Is a residue of 3 COOHF. In some embodiments, X 6 Is a residue of TfeGA. In some embodiments, X 6 Is an Asp residue. In some embodiments, X 6 Is [ CH2CMe2CO2H ]]Residues of TriAzDap. In some embodiments, X 6 Is a residue of Glu. In some embodiments, X 6 Is a residue of 2OH3 COOHF. In some embodiments, X 6 Is a residue of 4OH3 COOHF. In some embodiments, X 6 Is a residue of 4 COOHF. In some embodiments, X 6 Is a residue of 2 COOHF. In some embodiments, X 6 Is a residue of 5F3Me2 COOHF. In some embodiments, X 6 Is a residue of 4F3Me2 COOHF. In some embodiments, X 6 Is a residue of 5F3Me3 COOHF. In some embodiments, X 6 Is a residue of 4F3Me3 COOHF. In some embodiments, X 6 Is a residue of 3F2 COOHF. In some embodiments, X 6 Is a residue of dGlu.
In some embodiments, X 6 Is the residue of an amino acid whose side chain contains a polar group. Some are useful for X 6 Such amino acid residues of (a) include those directed against, for example, X 2 、X 5 And those described as having a polar group in a side chain thereof. In some embodiments, X 6 Is the residue of an amino acid whose side chain contains-OH. For example, in some embodiments, X 6 Is a residue of Thr, tyr, ser, aThr or hTyr. In some embodiments, X 6 Is the residue of Thr. In some embodimentsIn the scheme, X 6 Is a residue of Tyr. In some embodiments, X 6 Is a residue of Ser. In some embodiments, X 6 Is a residue of aThr. In some embodiments, X 6 Is a residue of hTyr. In some embodiments, X 6 Is a compound whose side chain contains an amide group (e.g., -C (O) N (R') 2 For example-CONH 2 ) Residues of amino acids of (a). In some embodiments, X 6 Is a residue of Asn. In some embodiments, X 6 Is Me2Gln.
In some embodiments, X 6 Is a residue of an amino acid whose side chain is hydrophobic. Some such amino acid residues include those directed against, for example, X 3 Those hydrophobic amino acid residues. In some embodiments, X 6 Is a residue of an amino acid whose side chain is an optionally substituted aliphatic group. In some embodiments, X 6 Is a residue of Val. In some embodiments, X 6 Is a residue of Ala. In some embodiments, X 6 Is a residue of Leu. In some embodiments, X 6 Is a residue of Ile.
Those skilled in the art who review this disclosure will readily appreciate that amino acid residues described for one location as having certain properties, structures, etc. may also be used in other locations where amino acid residues having the same properties, structures, etc. may be used. For example, when the hydrophobic amino acid residue may be at X 3 And X 6 When used in two positions, for X 3 The hydrophobic amino acid residues are useful in X 6 And vice versa. Similarly, when an acidic amino acid residue may be at X 2 、X 5 And X 6 When used at positions, the acidic amino acid residues described for one of them may also be used at the other two positions.
In some embodiments, X 6 Comprising a side chain containing an optionally substituted aromatic group. Some such amino acid residues include, for example, those directed against X 3 Those side chains contain amino acid residues of aromatic groups. In some embodiments, the aromatic group is optionally substituted having 1 to 3 nitrogen atoms5 membered heteroaryl. In some embodiments, the aromatic group is an optionally substituted 8-to 10-membered bicyclic aryl or heteroaryl group having 1 to 5 heteroatoms. In some embodiments, the aromatic group is phenyl. In some embodiments, the aromatic group is an optionally substituted phenyl group. In some embodiments, X 6 Is a residue of His. In some embodiments, X 6 Is a residue of Trp. In some embodiments, X 6 Is a residue of Phe. In some embodiments, X 6 Is a residue of 3cbmf.
In some embodiments, X 6 Is a residue selected from the group consisting of: 3COOHF, tfeGA, asp, [ CH2CMe2CO2H ]]TriAzDap, glu,2OH3COOHF, 4COOHF,2COOHF,5F3Me2COOHF,4F3Me2COOHF,5F3Me3COOHF,4F3Me3COOHF,3F2COOHF, dGlu, thr, tyr, ser, aThr, hTyr, glyn, lys, arg, val, ala, leu, phe, ile, his, trp, or 3cbmf.
In some embodiments, X 6 Is a residue of Gln. In some embodiments, X 6 Is a residue of Lys. In some embodiments, X 6 Is the residue of Arg.
In some embodiments, X 6 Is 3COOHF, asp, tfeGA, aib, glu, npg, gln, [ CH2CMe2CO2H ]]TriAzDap，B5，Thr，Ser，Asn，Ala，Hse，4BOH2F，2OH3COOHF，4OH3COOHF，4COOHF，2COOHF，His，Tyr，5F3Me2COOHF，4F3Me2COOHF，5F3Me3COOHF，4F3Me3COOHF，3F2COOHF，Val，Trp，Arg，dGlu，aThr，hTyr，3cbmf，Leu，Phe，Lys，Ile，SbMeAsp，bMe2Asp，3BOH2F，[Ac]Dap，[CH2CO2H[Acp，[Pfbn]GA，[Tfb]GA, succinic acid ester]Dap [ malonate]Dap，|Me2Mal]Dap，[SaiPrSuc]Dap，[SaMeSuc]Dap, or [ RaiPrSuc ]]Dap。
In some embodiments, X 6 Is 3COOHF. In some embodiments, X 6 Is Asp. In some embodiments, X 6 Is TfeGA. In some embodiments, X 6 Is Glu. In some embodiments, 3COOHF provides better properties and/or activity than, for example, asp.
In some embodiments, X 6 Is made ofAmino acid residues that are stapled, as described herein. In some embodiments, X 6 Is stapled. In some embodiments, X 6 Is the residue of B5.
In some embodiments, X 6 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-IV.
In some embodiments, X 6 Interact with Tyr306 of β -catenin or its corresponding amino acid residue. In some embodiments, X 6 Interacting with Lys345 of β -catenin or its corresponding amino acid residue.
Various types of amino acid residues can be used for X 7 For example, amino acids of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or residues of salts thereof, according to the present disclosure. In some embodiments, X 7 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 7 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 7 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H.
In some embodiments, R a2 Is R, wherein R is C 1-10 Aliphatic series. In some embodiments, R a3 Is R, wherein R is C 1-10 Aliphatic series. In some embodiments, R a2 And R is a3 Each independently is R, as described herein. In some embodiments, R a2 And R is a3 Are identical. In some embodiments, R is C 1-10 An alkyl group. In some embodiments, R is methyl.
In some embodiments, X 7 Is a residue of an amino acid whose side chain is hydrophobic. In some embodiments, X 7 Are hydrophobic amino acid residues as described herein, e.g., directed toX 3 Those described. In some embodiments, X 7 Is C whose side chain is optionally substituted 1-10 Residues of amino acids of alkyl groups. In some embodiments, X 7 Is that its side chain is C 1-10 Residues of amino acids of alkyl groups. In some embodiments, X 7 Is C whose side chain is optionally substituted by one or more non-polar and non-charged groups 1-10 Residues of amino acids of alkyl groups. In some embodiments, X 7 Comprising a side chain containing a cycloaliphatic group (e.g., a 3, 4, 5, or 6 membered cycloalkyl group). In some embodiments, X 7 Is a residue of Aib, ala, nLeu, cha, npg, sAla, val, cyLeu, leu, aMeL, daMeL or aMeV. In some embodiments, X 7 Is Aib, ala, nLeu or Cha. In some embodiments, X 7 Are residues of Aib. In some embodiments, X 7 Is a residue of Ala. In some embodiments, X 7 Is a residue of nLeu. In some embodiments, X 7 Is the residue of Cha. In some embodiments, X 7 Is a residue of Npg. In some embodiments, X 7 Is a residue of sA. In some embodiments, X 7 Is a residue of Val. In some embodiments, X 7 Is a residue of CyLeu. In some embodiments, X 7 Is a residue of Leu. In some embodiments, X 7 Is a residue of Cpg. In some embodiments, X 7 Is a residue of Cbg. In some embodiments, X 7 Is a residue of aMeL. In some embodiments, X 7 Is a residue of DaMeL. In some embodiments, X 7 Is a residue of aMeV.
In some embodiments, X 7 Is the residue of an amino acid whose side chain contains a polar group. A variety of polar amino acid residues as described herein can be used for X 7 . In some embodiments, X 7 Is the residue of an amino acid whose side chain contains-OH. For example, in some embodiments, X 7 Is a residue of Ser. In some embodiments, X 7 Is a residue of Hse. In some embodiments, X 7 Is the residue of Thr. In some implementationsIn embodiments, X 7 Are residues of DaMeS. In some embodiments, X 7 Is a residue of aMeS.
In some embodiments, X 7 Is the residue of an amino acid comprising an acidic or polar group. In some embodiments, X 7 Is a residue of an amino acid whose side chain contains an acidic group (e.g., -COOH group) or a salt form thereof (e.g., a compound of formula a-IV, etc.). A variety of acidic amino acid residues described herein can be used for X 7 . In some embodiments, X 7 Is a residue of 3 COOHF. In some embodiments, X 7 Is a compound whose side chain contains an amide group (e.g., -C (O) N (R') 2 For example-CONH 2 ) Residues of amino acids of (a). In some embodiments, X 7 Is a residue of Asn. In some embodiments, X 7 Is a residue of Gln. In some embodiments, X 7 Is a residue of Me2 Gln. In some embodiments, X 7 Is a residue of AcLys.
In some embodiments, X 7 Comprising a side chain containing an optionally substituted aromatic group. A variety of aromatic amino acid residues as described herein can be used for X 7 . In some embodiments, the aromatic group is an optionally substituted 5-membered heteroaryl group having 1 to 3 nitrogen atoms. In some embodiments, X 7 Is a residue of Phe. In some embodiments, X 7 Is a residue of aMeDF. In some embodiments, X 7 Is a residue of aMeF. In some embodiments, X 7 Is a residue of His.
In some embodiments, X 7 Selected from Aib, ala, morphGln, gln, glnR, ser, iPrLys, nLeu, cha, hse, lys, npg, sAla, triAzLys, val, cyLeu, 3COOHF, thr, phe, [29N2 spiroundecane]GlnR, acp, asn, daMeS, aMeDF, [4 Aminopiperidines]GlnR, leu, cpg, cbg, me2Gln, met2O, acLys, his, aMeL, daMeL, aMeV, aMeS, aMeF, [ isophthalate ] ]Lys, [ succinic acid ester ]]Lys、[Me2Mal]Lys, [ dibenzoate ]]Lys([diphenate]Lys) or [ biphenyl 33COOH]Lys. In some embodiments, X 7 Selected from GlnR, lys, [29N2 spiroundecane]GlnR、[4 Aminopiperidines]GlnR, sAla, triAzLys, [ isophthalate ]]Lys, [ succinic acid ester ]]Lys、[Me2Mal]Lys, [ dibenzoate ]]Lys or [ biphenyl 33COOH]Lys。
In some embodiments, X 7 Are suitable amino acid residues for stapling, as described herein.
In some embodiments, the amino acid residue suitable for stapling is-N (R a1 )-L a1 -C(-L a -R SP1 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, it is-N (R a1 )-C(-L a -R SP1 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, each amino acid residue is independently-N (R a1 )-L a1 -C(-L a -R SP1 )(R a3 )-L a2 -C (O) -or-N (R) a1 )-C(-L a -R SP1 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H. In some embodiments, R a1 And R is a3 Both are-H.
In some embodiments, R is an amino acid residue in a pair SP1 is-NHR, wherein R is as described herein. In some embodiments, R is-H. In some embodiments, R is optionally substituted C 1-6 Aliphatic series. In some embodiments, R is optionally substituted C 1-6 An alkyl group. In some embodiments, R is C 1-6 Aliphatic series. In some embodiments, R is C 1-6 An alkyl group. In some embodiments, R SP1 is-NH 2 . In some embodiments, such amino acid residues may be stapled to another amino acid residue comprising-COOH by amidation to form a staple-like structure comprising-C (O) N (R') -, e.g., L s Wherein L is s2 is-C (O) N (R ') -or comprises-C (O) N (R') -. In some embodiments, at another amino acid residue of the pairR in (B) SP1 is-COOH or an active derivative thereof. In some embodiments, R in another amino acid residue of a pair SP1 is-COOH. In some embodiments, R' is R. In some embodiments, R' is-H. In some embodiments, L s1 Is a first amino acid residue (e.g.X 7 ) L of (2) a . In some embodiments, L s3 L being a second amino acid residue (e.g. the C-direction amino acid residue of the first amino acid residue) a . In some embodiments, the first amino acid residue is X 7 And the second amino acid residue is X 7 C-oriented amino acid residues of (2), e.g. X 10 . In some embodiments, L s1 And L s3 Each independently is L. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In some embodiments, L s1 And L s3 Each independently is L, wherein L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L s1 And L s3 Each independently is L, wherein L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L s -(CH 2 )n1-C(O)N(R’)-(CH 2 ) n2 -wherein each variable is independently as described herein. In some embodiments, n1 and n2 are each independently 1 to 10. In some embodiments, the first amino acid residue has R as an amino group SP1 And the second amino acid residue has R as-COOH or an activated form thereof SP1 . In some embodiments, the second amino acid residue has R as an amino group SP1 And the first amino acid residue has R as-COOH or an activated form thereof SP1 . In some embodiments, the first amino acid residue is X 7 And the second amino acid residue is one of its C-direction amino acid residues, e.g. X 10 . In some embodiments, the second amino acid residue is X 7 And the first amino acid residue is one of its N-directional amino acid residues, e.g. X 3 . In some embodiments, the first amino acid residue is X 7 . In some embodiments, X 7 Is Lys. In some embodiments, the second amino acid residue is X 10 . In some embodiments, X 10 Is GlnR. In some embodiments, n1 is 4, as in Lys. In some embodiments, n2 is 2, as in GlnR. In some embodiments, the first amino acid residue is X 7 Such as GlnR. In some embodiments, n1 is 2. In some embodiments, the second amino acid residue is X 10 Such as Lys. In some embodiments, n2 is 4. In some embodiments, the second amino acid residue is(e.g., X 14 ). In some embodiments, L s3 Is- (CH) 2 ) 2 -C(O)NH-(CH 2 ) 4 -. In some embodiments, the second amino acid residue is(e.g., X 14 ). In some embodiments, L s3 Is- (CH) 2 ) 2 -C (O) -Cy-. In some embodiments, -Cy-is optionally substituted +.>Wherein nitrogen is bonded to-C (O) -. In some embodiments, L s3 Is- (CH) 2 ) 2 -C(O)-N(R’)-(CH 2 ) n-CHR '-, in which two R's together with intervening atoms form an optionally substituted ring, as described herein. In some embodiments, the ring formed is optionally substituted +.>In some embodiments, the second amino acid residue is +.>(e.g., X 14 ). In some embodiments, L s3 Is- (CH) 2 ) 2 -C(O)-N(R’)-(CH 2 ) n-Cy-. In some embodiments, R' is R, as described herein. In some embodiments, R is-H. In some embodiments, R is optionally substituted C 1-6 Aliphatic series. In some embodiments, R is optionally substituted C 1-6 An alkyl group. In some embodiments, R is methyl. In some embodiments, n is 1. In some embodiments, -Cy-is optionally substituted +. >Wherein nitrogen is with L s2 Bonding, L s2 is-C (O) -or comprises-C (O) -. In some embodiments, L s3 Is- (CH) 2 ) 2 -C(O)-N(R’)-CH 2 -CHR’-(CH 2 ) n-. In some embodiments, n is 2. In some embodiments, - (CH) 2 ) n-and L s2 -N (R') -linkage, L s2 is-C (O) -N (R') -. In some embodiments, L s3 R 'and L of-CHR' -, are described s2 R 'of-N (R') -and intervening atoms thereof taken together formAn optionally substituted ring, as described herein. In some embodiments, the ring formed is optionally substituted +.>In some embodiments, the second amino acid residue is +.>(e.g., X 14 ). In some embodiments, the second amino acid residue is +.>(e.g., X 14 ). In some embodiments, L s3 Is- (CH) 2 ) 2 -C(O)-N(R’)-(CH 2 ) n1 -C(R’) 2 -(CH 2 ) n2 -. In some embodiments, n1 and n2 are each independently 1 to 10. In some embodiments, n1 is 1. In some embodiments, n1 is 2. In some embodiments, n2 is 2. In some embodiments, -N (R '), -R ' and-C (R ') 2 -one R' taken together with the intervening atoms thereof forms an optionally substituted ring, as described herein. In some embodiments, the ring formed is an optionally substituted 6 membered monocyclic saturated ring, which ring has no heteroatoms other than the nitrogen atom of-N (R') -. In some embodiments, L s2 is-C (O) N (R') -. In some embodiments, -N (R'), -and- (CH) 2 ) n2 -bonding. In some embodiments, L s3 Is (R') 2 -one R' and L s2 R 'of-N (R') -and intervening atoms thereof taken together form an optionally substituted ring, as described herein. In some embodiments, the ring formed is an optionally substituted 6 membered monocyclic saturated ring, which ring has no heteroatoms other than the nitrogen atom of-N (R') -.
In some embodiments, the first amino acid residue is(e.g., X 7 ). In one placeIn some embodiments, L s1 Is- (CH) 2 ) 2 -C(O)-N(R’)-(CH 2 ) n-CHR '-, in which two R's together with intervening atoms form an optionally substituted ring, as described herein. In some embodiments, the ring formed is optionally substitutedIn some embodiments, the second amino acid residue is GlnR (e.g., X 14 ). In some embodiments, L s3 Is- (CH) 2 ) 2 -。
In some embodiments, the first amino acid residue is(e.g., X 7 ). In some embodiments, L s1 Is- (CH) 2 ) 2 -C(O)-N(R’)-(CH 2 ) n-Cy-. In some embodiments, R' is R, as described herein. In some embodiments, R is-H. In some embodiments, R is optionally substituted C 1-6 Aliphatic series. In some embodiments, R is optionally substituted C 1-6 An alkyl group. In some embodiments, R is methyl. In some embodiments, n is 1. In some embodiments, -Cy-is optionally substituted +.>Wherein nitrogen is with L s2 Bonding, L s2 is-C (O) -or comprises-C (O) -. In some embodiments, L s1 Is- (CH) 2 ) 2 -C(O)-N(R’)-CH 2 -CHR’-(CH 2 ) n-. In some embodiments, n is 2. In some embodiments, - (CH) 2 ) n-and L s2 -N (R') -linkage, L s2 is-C (O) -N (R') -. In some embodiments, L s1 R 'and L of-CHR' -, are described s2 R 'of-N (R') -and intervening atoms thereof taken together form an optionally substituted ring, as described herein. In some embodiments, the ring formed is optionally substitutedIn some embodiments, the first amino acid residue is +.>(e.g., X 7 ). In some embodiments, the first amino acid residue is +.>(e.g., X 7 ). In some embodiments, L s1 Is- (CH) 2 ) 2 -C(O)-N(R’)-(CH 2 ) n1 -C(R’) 2 -(CH 2 ) n2 -. In some embodiments, n1 and n2 are each independently 1 to 10. In some embodiments, n1 is 1. In some embodiments, n1 is 2. In some embodiments, n2 is 2. In some embodiments, -N (R '), -R ' and-C (R ') 2 -one R' taken together with the intervening atoms thereof forms an optionally substituted ring, as described herein. In some embodiments, the ring formed is an optionally substituted 6 membered monocyclic saturated ring, which ring has no heteroatoms other than the nitrogen atom of-N (R') -. In some embodiments, L s2 is-C (O) N (R') -. In some embodiments, -N (R'), -and- (CH) 2 ) n2 -bonding. In some embodiments, L s1 Is (R') 2 -one R' and L s2 R 'of-N (R') -and intervening atoms thereof taken together form an optionally substituted ring, as described herein. In some embodiments, the ring formed is an optionally substituted 6 membered monocyclic saturated ring, which ring has no heteroatoms other than the nitrogen atom of-N (R') -. In some embodiments, the second amino acid residue is GlnR (e.g., X 14 )。
In some embodiments, the first residue is(e.g., X 7 ). In some embodiments, the first residue is +.>(e.g., X 7 ). In some embodiments, the first residue is +.>(e.g., X 7 ). In some embodiments, L s1 Is- (CH) 2 ) N-N (R') -C (O) -Cy-Cy-, wherein each variable is independently as described herein. In some embodiments, L s1 Is- (CH) 2 ) N-N (R') -C (O) -Cy-, wherein each variable is independently as described herein. In some embodiments, the first residue is +.>(e.g., X 7 ). In some embodiments, L s1 Is- (CH) 2 )n-N(R’)-C(O)-CH 2 -, wherein R is as described herein and is bonded to C (O) -, a-CH 2 -is optionally substituted. In some embodiments, L s1 Is- (CH) 2 )n-N(R’)-C(O)-C(R’) 2 -, wherein each R is independently as described herein. In some embodiments, L s1 Is- (CH) 2 )n-N(R’)-C(O)-C(CH 3 ) 2 -, wherein R is as described herein. In some embodiments, the first residue is +.>(e.g., X 7 )。
In some embodiments, L s1 Is- (CH) 2 ) n1 -N(R’)-C(O)-(CH 2 ) n2 -wherein each variable is independently as described herein. In some embodiments, n1 and n2 are each independently n, as described herein. In some embodiments, L s1 Is- (CH) 2 ) 4 -N(R’)-C(O)-(CH 2 ) 2 -, wherein each R is independently as described herein. In some embodiments, n is 1 to 10. In some embodiments, n is 1. In some embodiments, n is 2. At the position ofIn some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, R' is R, as described herein. In some embodiments, R is-H. In some embodiments, -Cy-is optionally substituted phenylene. In some embodiments, -Cy-is optionally substituted 1, 2-phenylene. In some embodiments, -Cy-is optionally substituted 1, 3-phenylene. In some embodiments, each-Cy-is independently optionally substituted 1, 2-phenylene. In some embodiments, each-Cy-is independently optionally substituted 1, 3-phenylene. In some embodiments, L s2 is-C (O) -N (R ') -or comprising-C (O) -N (R') -, as described herein. In some embodiments, R' is R, as described herein. In some embodiments, R is-H. In some embodiments, L s2 is-C (O) NH-. In some embodiments, -C (O) -and L s1 -Cy-bond of (c). In some embodiments, the second residue is X 14 Such as Lys. In some embodiments, L s3 Is for example optionally substituted- (CH) as described herein 2 ) n-. In some embodiments, L s3 Is- (CH) 2 ) n-. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4 (e.g., as in Lys).
In some embodiments, R of the first amino acid residue SP1 Is or comprises-COOH or a protected or activated form thereof. In some embodiments, the first amino acid residue is X 3 Such as GlnR. In some embodiments, R of the second amino acid residue SP1 Is or comprises the following: amino, e.g., -NHR as described herein. In some embodiments, R of the second amino acid residue SP1 is-NH 2 Or comprises-NH 2 . In some embodiments, the second amino acid residue is X 7 Such as Lys. In some embodiments, L s1 And L s3 Each independently is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L s1 Is- (CH) 2 ) 2 -. In some embodiments, L s1 Is- (CH) 2 ) 4 -。
In some embodiments, R is an amino acid residue in a pair SP1 Is the first reactive group of the cycloaddition reaction. In some embodiments, such an amino acid residue may be stapled by a cycloaddition reaction with another amino acid residue comprising a second reactive group of the cycloaddition reaction. In some embodiments, R in another amino acid residue of a pair SP1 Is the second reactive group of the cycloaddition reaction. In some embodiments, the cycloaddition reaction is [3+2 ]]. In some embodiments, the cycloaddition reaction is a click chemistry reaction. In some embodiments, the cycloaddition reaction is [4+2 ]]. In some embodiments, one of the first and second reactive groups is-N 3 Or comprises-N 3 And the other is or comprises an alkyne (e.g., a terminal alkyne or an activated/strained alkyne).
In some embodiments, R of the first amino acid residue SP1 is-N 3 . In some embodiments, L of the first amino acid residue a Is L, as described herein. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -, -C (O) -, -N (R')-, -Cy-, or-O-substitution. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution.
In some embodiments, R of the second amino acid residue SP1 is-C.ident.C-or contains-C.ident.C-. In some embodiments, R of the second amino acid residue SP1 Is thatIn some embodiments, R of the second amino acid residue SP1 Including, for example, strained alkynes in the ring. In some embodiments, L of the first amino acid residue a Is L, as described herein. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution.
In some embodiments, L s is-L s1 -L s2 -L s3 -, wherein L s2 is-Cy-orcomprises-Cy-. In some embodiments, L s2 is-Cy-. In some embodiments, -Cy-is formed by a cycloaddition reaction. In some embodiments, -Cy-is optionally substitutedIn some embodiments, -Cy-is +.>In some embodiments, -Cy-is formed by a cycloaddition reaction. In some embodiments, -Cy-is optionally substituted +.>In some embodiments, -Cy-is +.>In some embodiments, L s1 L being the first amino acid residue a And L is s3 L being a second amino acid residue a . In some embodiments, L s1 L being a second amino acid residue a And L is s3 L being the first amino acid residue a . In some embodiments, L s1 And L s3 Each independently is L, as described herein. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodimentsIn this case, L is an optionally substituted divalent straight-chain or branched C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In some embodiments, L s1 Is optionally substituted- (CH) 2 ) n -, wherein n is 1 to 10. In some embodiments, L s1 Is- (CH) 2 ) n -, wherein n is 1 to 10. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, L s3 Is optionally substituted- (CH) 2 ) n -, wherein n is 1 to 10. In some embodiments, L s3 Is- (CH) 2 ) n -, wherein n is 1 to 10. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4.
In some embodiments, the first amino acid residue is X 7 . In some embodiments, X 7 R of (2) SP1 is-N 3 . In some embodiments, X 7 L of (2) a Is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, X 7 L of (2) a Is- (CH) 2 ) 4 -. In some embodiments, X 7 L of (2) a Is- (CH) 2 ) 3 -. In some embodiments, X 7 L of (2) a Is- (CH) 2 ) 2 -. In some embodiments, X 7 L of (2) a is-CH 2 -. In some embodiments, the second amino acid residue is X 10 . In some embodiments, X 10 R of (2) SP1 Is or comprises an alkyne, for example, a strained/activated alkyne. In some embodiments, X 10 R of (2) SP1 is-C.ident.CH. In some embodiments, X 10 L of (2) a Is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, X 10 L of (2) a Is- (CH) 2 ) 4 -. In some embodiments, X 10 L of (2) a Is- (CH) 2 ) 3 -. In some embodiments, X 10 L of (2) a Is- (CH) 2 ) 2 -. In some embodiments, X 10 L of (2) a is-CH 2 -. In some embodiments, L s3 Is X 10 L of (2) a . In some embodiments, L s3 And L is equal to s2 Is bonded to a carbon atom of (c).
In some embodiments, the first amino acid residue is X 7 . In some embodiments, X 7 R of (2) SP1 Is or comprises an alkyne, for example, a strained/activated alkyne. In some embodiments, X 7 R of (2) SP1 is-C.ident.CH. In some embodiments, X 7 L of (2) a Is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, X 7 L of (2) a Is- (CH) 2 ) 4 -. In some embodiments, X 7 L of (2) a Is- (CH) 2 ) 3 -. In some embodiments, X 7 L of (2) a Is- (CH) 2 ) 2 -. In some embodiments, X 7 L of (2) a is-CH 2 -. In some embodiments, L s1 Is X 7 L of (2) a . In some embodiments, L s1 And L is equal to s2 Is bonded to a carbon atom of (c). In some embodiments, the second amino acid residue is X 10 . In some embodiments, X 10 R of (2) SP1 is-N 3 . In some embodiments, X 10 L of (2) a Is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, X 10 L of (2) a Is- (CH) 2 ) 4 -. In some embodiments, X 10 L of (2) a Is- (CH) 2 ) 3 -. In some embodiments, X 10 L of (2) a Is- (CH) 2 ) 2 -. In some embodiments, X 10 L of (2) a is-CH 2 -。
In some embodiments, R of two amino acid residues of a pair of amino acid residues suitable for stapling SP1 Can each independently react with a linking agent to form a staple-like structure. In some embodiments, suitable linking reagents comprise two reactive groups, each of which may be independently associated with R of each amino acid residue SP1 And (3) reacting. In some embodiments, the linking agent has an H-L "-H structure or salt thereof, wherein the agent comprises two amino groups, and L" is a covalent bond or an optionally substituted divalent C 1 -C 20 An aliphatic group wherein one or more methylene units of the aliphatic group are optionally and independently substituted
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 N (R') -C (O) S-, or-C (O) O-. In some embodiments, such linking reagents can react with two amino acid residues, each independently having R SP1 A group which is-COOH or an activated form thereof.
Some suitable embodiments of L "include those described herein for L that fall within the scope of L". For example, in some embodiments, L "is L, wherein L is an optionally substituted divalent straight or branched chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is- (CH) 2 ) n-, where nIs 1 to 10. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution.
In some embodiments, the linking agent is a diamine or salt thereof. In some embodiments, the agent has a NHR-L "-NHR structure or a salt thereof, wherein each variable is independently as described herein. In some embodiments, each R is independently-H or optionally substituted C 1-6 Aliphatic series. In some embodiments, each R is independently-H or C 1-6 Aliphatic series. In some embodiments, each R is independently-H or optionally substituted C 1-6 An alkyl group. In some embodiments, each R is independently-H or C 1-6 An alkyl group. In some embodiments, the reagent has NH 2 -L”-NH 2 A structure or a salt thereof. In some embodiments, L "is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L "is- (CH) 2 ) 4 -。
In some embodiments, staple-like structure L s is-L s1 -L s2 -L s3 -, wherein L s1 L being the first amino acid residue of a stapled pair a ，L s3 L being the second amino acid residue of a stapled pair a And L is s2 is-C (O) -N (R ') -L "-N (R') -C (O) -, wherein each variable is independently as described herein. In some embodiments, L "is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L "is- (CH) 2 ) 4 -. In some embodiments, L s1 And L s3 Each independently is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, n is 2. In some embodiments, the first amino acid residueThe radical being Gln (e.g. X 7 ). In some embodiments, the second amino acid residue is GlnR (e.g., X 14 ). In some embodiments, two GlnRs can be obtained by [ diaminobutane]Such a staple-like structure is formed.
In some embodiments, the linking reagent has an H-Cy-L "-NHR structure or salt thereof, wherein-Cy-comprises a second amino group. In some embodiments, R is-H or optionally substituted C 1-6 Aliphatic series. In some embodiments, R is-H or C 1-6 Aliphatic series. In some embodiments, R is-H or optionally substituted C 1-6 An alkyl group. In some embodiments, R is-H or C 1-6 An alkyl group. In some embodiments, R is methyl. In some embodiments, the linking reagent has H-Cy-L "-NH 2 A structure or a salt thereof, wherein-Cy-comprises a second amino group. In some embodiments, -Cy-is optionally substitutedIn some embodiments, -Cy-is +.>In some embodiments, L "is a covalent bond. In some embodiments, L "is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L "is- (CH) 2 ) -. In some embodiments, the linking agent is +.>Or a salt thereof. In some embodiments, the linking agent isOr a salt thereof.
In some embodiments, L s2 is-C (O) -Cy-N (R') -C (O) -, wherein each variable is independently as described herein. In some embodiments, R' is-H. In some embodiments, -Cy-isIn some embodiments, L s1 And L s3 Each independently is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, n is 2. In some embodiments, -Cy-is closer to the N-terminus than-N (R') -. In some embodiments, -Cy-is closer to the C-terminus than-N (R') -. In some embodiments, the first amino acid residue is Gln (e.g., X 7 ). In some embodiments, the second amino acid residue is GlnR (e.g., X 14 ). In some embodiments, two GlnRs can be substituted by [4 aminopiperidines]Such a staple-like structure is formed.
In some embodiments, L s2 is-C (O) -Cy- (CH) 2 ) N-N (R') -C (O) -, wherein each variable is independently as described herein. In some embodiments, R' is-H. In some embodiments, R' is R, as described herein, e.g., optionally substituted C 1-6 Aliphatic, C 1-6 Alkyl groups, and the like. In some embodiments, R is methyl. In some embodiments, n is 1. In some embodiments, -Cy-isIn some embodiments, -Cy-is closer to the N-terminus than-N (R') -. In some embodiments, -Cy-is closer to the C-terminus than-N (R') -. In some embodiments, L s1 And L s3 Each independently is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, n is 2. In some embodiments, the first amino acid residue is Gln (e.g., X 7 ). In some embodiments, the second amino acid residue is GlnR (e.g., X 14 ). In some embodiments, two GlnRs can be obtained by [4 mamplatidine ]]Such a staple-like structure is formed.
In some embodiments, the methylene units are replaced with-Cy-. In some embodiments, the linking reagent has an H-Cy-H structure, wherein Cy comprises two secondary amino groups. In some embodiments, -Cy-is an optionally substituted 8-to 20-membered bicyclic ring. In some embodiments, H-Cy-H comprises two-NH-. In some casesIn embodiments, -Cy-is optionally substitutedIn some embodiments, -Cy-is optionally substituted +.>In some embodiments, the meta-position linkage site (relative to the spiro carbon atom) is closer to the N-terminus than the para-position linkage site (relative to the spiro carbon atom). In some embodiments, the meta-position linkage site (relative to the spiro carbon atom) is closer to the C-terminus than the para-position linkage site (relative to the spiro carbon atom).
In some embodiments, L s2 is-C (O) -Cy-C (O) -, wherein-Cy-is as described herein. In some embodiments, L s1 And L s3 Each independently is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, n is 2. In some embodiments, the first amino acid residue is Gln (e.g., X 7 ). In some embodiments, the second amino acid residue is GlnR (e.g., X 14 ). In some embodiments, two GlnRs can be obtained by reacting [29N2 spiroundecane ]Such a staple-like structure is formed. In some embodiments, two GlnRs can be obtained by reacting [39N2 spiroundecane]Such a staple-like structure is formed.
In some embodiments, the pairs of amino acid residues suitable for stapling both independently have-N (R a1 )-L a1 -C(-L a -R SP1 )(R a3 )-L a2 -C (O) -or-N (R) a1 )-C(-L a -R SP1 )(R a3 ) -C (O) -structure, wherein each variable is independently as described herein, and R SP1 Is an amino group. In some embodiments, R SP1 is-NHR, wherein R is as described herein. In some embodiments, R is-H. In some embodiments, R is optionally substituted C 1-6 Aliphatic series. In some embodiments, R is optionally substituted C 1-6 An alkyl group. In some embodiments, R is C 1-6 Aliphatic series. In some embodiments, R isC 1-6 An alkyl group. In some embodiments, R SP1 is-NH 2 . In some embodiments, such two amino acid residues may be linked by a diacid linking reagent.
In some embodiments, the linking agent has the HOOC-L "-COOH structure or a salt or activated form thereof, wherein L" is as described herein. In some embodiments, L "is-Cy-. In some embodiments, L "is-Cy-. In some embodiments, -Cy-is optionally substituted phenylene. In some embodiments, -Cy-is optionally substituted 1, 2-phenylene. In some embodiments, -Cy-is optionally substituted 1, 3-phenylene. In some embodiments, each-Cy-is independently optionally substituted 1, 2-phenylene. In some embodiments, each-Cy-is independently optionally substituted 1, 3-phenylene. In some embodiments, L "is optionally substituted In some embodiments, the linking agent is +.>Or a salt or activated form thereof. In some embodiments, L "is optionally substituted +.>In some embodiments, the linking agent is +.>Or a salt or activated form thereof. In some embodiments, L' is 1, 3-phenylene. In some embodiments, the linking agent isOr a salt or activated form thereof. In some embodiments, L "is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L' is an optionally substituted-CH 2 -. In some embodiments, L 'is-C (R') 2 -. In some embodiments, L' is-C (CH 3 ) 2 -. In some embodiments, the linking agent is (CH 3 ) 2 C(COOH) 2 Or a salt or activated form thereof. In some embodiments, L' is-CH 2 CH 2 -. In some embodiments, the linking agent is hoogch 2 CH 2 COOH or a salt or activated form thereof.
In some embodiments, the staple-like structure is L s Wherein L is s2 is-N (R ') -L ' -N (R ') -and L s1 And L s3 Each independently as described herein. In some embodiments, L "is-Cy-, wherein each-Cy-is independently as described herein. In some embodiments, L "is-Cy-, as described herein. In some embodiments, -Cy-is optionally substituted phenylene. In some embodiments, -Cy-is optionally substituted 1, 2-phenylene. In some embodiments, -Cy-is optionally substituted 1, 3-phenylene. In some embodiments, each-Cy-is independently optionally substituted 1, 2-phenylene. In some embodiments, each-Cy-is independently optionally substituted 1, 3-phenylene. In some embodiments, L "is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L' is an optionally substituted-CH 2 -. In some embodiments, L 'is-C (R') 2 -. In some embodiments, L' is-C (CH 3 ) 2 -. In some embodiments, L' is-CH 2 CH 2 -. In some embodiments, L s1 And L s3 Each independently is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, n is 2. In some embodiments, n is 4. In some embodiments, the first amino acid residue is Lys (e.g., X 7 ). In some embodiments, the second amino acid residue is Lys (e.g., X 14 ). In some embodiments, two lyss may pass through [ biphenyl 33COOH]Such a staple-like structure is formed. In some embodiments, two lyss may be linked by [ dibenzoate]Such a staple-like structure is formed. In some embodiments of the present invention, in some embodiments,two Lyss can pass through [ isophthalate]Such a staple-like structure is formed. In some embodiments, two lyss may be linked by [ Me2Mal]Such a staple-like structure is formed. In some embodiments, two Lys may be performed by [ succinate]Such a staple-like structure is formed.
In some embodiments, X 7 Is stapled. In some embodiments, X 7 And X is 14 Stapling. In some embodiments, X 7 And X is 10 Stapling. In some embodiments, X 10 And X is 7 Stapling. In some embodiments, X 7 And X is 3 Stapling.
In some embodiments, X 7 Is Aib, ala, 3COOHF, cyLeu, phe, asp, nLeu, B5, val, gln, morphGln, glnR, cha, ser, leu, cbg, cyhLeu, iPrLys, aic, lys, lys, hse, glnR, npg, glnR, dpg, gly, sAla, triAzLys, thr, asn, dAla, [ isophthalate ]]Lys, [ succinic acid ester ]]-Lys, [29N2 spiroundecane]GlnR、Acp、DaMeS、aMeDF、DGlnR、[Ac]Acp、[Phc]Acp, [ isovaleryl ]]Acp、[Me2Mal]-Lys, [ dibenzoate ]]-Lys, [ biphenyl 33COOH]-Lys、[Me2Mal]Lys, [ dibenzoate ]]Lys, [ biphenyl 33COOH]Lys, [4 Aminopiperidine]GlnR, cpg, me2Gln, met2O, acLys, his, aMeL, daMeL, aMeV, aMeS, aMeF, dLys, [ ethylenediamine ]]GlnR, [ Me2 ethylenediamine ]]GlnR, [ diaminopropane ]]GlnR, [ diaminopentane ]]GlnR, [ Me2 diaminohexane ]]GlnR、[Ac]PyrSa、[Phc]PyrSa, [ isovaleryl ]]PyrSa、[Ac]PyrRa、[Phc]Pyrra, [ isovaleryl ]]PyrRA, 2COOHF, 4COOHF or Glu. In some embodiments, X 7 Is Aib. In some embodiments, X 7 Is Ala. In some embodiments, X 7 Is 3COOHF. In some embodiments, X 7 Is CyLeu. In some embodiments, X 7 Is Phe. In some embodiments, X 7 Is nLeu. In some embodiments, X 7 Is Val. In some embodiments, X 7 Is Cha. In some embodiments, X 7 Is Leu. In some embodiments, X 7 Is Cbg. In some embodimentsIn the case of X 7 Is CyhLeu. In some embodiments, aib provides better properties and/or activity than, for example, ala. In some embodiments, X 7 Is GlnPDA 3. In some embodiments, X 7 Is GlnBDA 3. In some embodiments, X 7 Is GlnR 3. In some embodiments, X 7 Is GlnMeBDA x 3. In some embodiments, X 7 Is GlnT4CyMe 3. In some embodiments, X 7 Is GlnC4CyMe 3. In some embodiments, X 7 Is Gln3 acppip 3. In some embodiments, X 7 Is GlnPipAz 3. In some embodiments, X 7 Is Gln4pipp 3. In some embodiments, X 7 Is GlnPip4AE 3.
In some embodiments, X 7 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-I, tables A-II, tables A-III and tables A-IV.
Various types of amino acid residues can be used for X 8 For example, amino acids of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or residues of salts thereof, according to the present disclosure. In some embodiments, X 8 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 8 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 8 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H.
In some embodiments, X 8 Is a residue of an amino acid whose side chain is hydrophobic. In some embodiments, X 8 Are hydrophobic amino acid residues as described herein, e.g., for X 3 Those described. In some embodiments, X 8 Is a residue of Ala. In some embodiments, X 8 Are residues of Aib. In some embodiments, X 8 Is a residue of Cpg. In some embodiments, X 8 Is a residue of Val. In some embodiments, X 8 Is a residue of Leu. In some embodiments, X 8 Is a residue of nLeu. In some embodiments, X 8 Is a residue of Cba.
In some embodiments, X 8 Is the residue of an amino acid comprising an acidic or polar group. In some embodiments, X 8 Is the residue of an amino acid whose side chain contains a polar group. In some embodiments, X 8 Are polar amino acid residues as described herein. In some embodiments, X 8 Is the residue of an amino acid whose side chain contains-OH. In some embodiments, X 8 Comprising a side chain containing an optionally substituted aromatic group. For example, in some embodiments, X 8 Is a residue of Ser. In some embodiments, X 8 Is the residue of Thr. In some embodiments, X 8 Is a residue of aThr. In some embodiments, X 8 Is a residue of hTyr. In some embodiments, X 8 Is a compound whose side chain contains an amide group (e.g., -C (O) N (R') 2 For example-CONH 2 ) Residues of amino acids of (a). In some embodiments, X 8 Is a residue of Gln. In some embodiments, X 8 Is a residue of AcLys.
In some embodiments, X 8 Are residues of amino acids whose side chains contain acidic groups (e.g., -COOH groups) or of their salt forms (e.g., compounds of formulae a-IV). In some embodiments, X 8 Are acidic amino acid residues as described herein, e.g., for X 2 、X 5 、X 6 Etc. as described. In some embodiments, X 8 Is an Asp residue. In some embodiments, X 8 Is a residue of Glu. In some embodiments, X 8 Is a residue of Aad.
In some embodiments, X 8 Comprising a side chain containing an optionally substituted aromatic group. In some embodiments, X 8 Are aromatic amino acid residues as described herein. In some embodiments, the aromatic group isPhenyl. In some embodiments, X 8 Is a residue of Phe. In some embodiments, X 8 Is the residue of hpe. In some embodiments, X 8 Is a residue of hTyr.
In some embodiments, X 8 Selected from Ala, aib, cpg, val, leu, gln, lys, asp, glu, aad, nLeu, cba, ser, thr, aThr, morphGLn, phe, hPHE, hTyr, and AcLys.
In some embodiments, X 8 Ala, aib, phe, asp,3COOHF, aThr, gly, ser, nLeu, thr, cpg, val, leu, gln, lys, glu, aad, cba, morphGLn, hPr, hTyr, or AcLys. In some embodiments, X 8 Is Ala. In some embodiments, X 8 Is Aib. In some embodiments, X 8 Is Phe. In some embodiments, X 8 Is Asp. In some embodiments, X 8 Is 3COOHF.
In some embodiments, X 8 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-IV.
In some embodiments, X 8 Interact with Trp383 of beta-catenin or its corresponding amino acid residue.
Various types of amino acid residues can be used for X 9 For example, amino acids of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or residues of salts thereof, according to the present disclosure. In some embodiments, X 9 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 9 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 9 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H.
In some embodiments, X 9 Comprising optionally substituted aromatic groupsSide chains of aromatic groups. In some embodiments, X 9 Are aromatic amino acid residues as described herein. In some embodiments, the aromatic group is an optionally substituted 5-membered heteroaryl group having 1 to 3 heteroatoms. In some embodiments, the aromatic group is an optionally substituted 5-membered heteroaryl group having 1 to 3 nitrogen atoms. In some embodiments, the aromatic group is an optionally substituted 5-membered heteroaryl group having one sulfur atom. In some embodiments, the aromatic group is an optionally substituted phenyl group. In some embodiments, X 9 Comprising a side chain which is OR comprises an optionally substituted aromatic group, wherein each substituent of the aromatic group is independently selected from halogen, -OR, -R, -C (O) OH OR-CN, wherein each R is independently hydrogen OR C 1-4 Alkyl or haloalkyl. In some embodiments, the aromatic group is phenyl. In some embodiments, the aromatic group is an optionally substituted 8-to 10-membered bicyclic aryl or heteroaryl having 1 to 5 heteroatoms. In some embodiments, X 9 Comprising a side chain which is or comprises an optionally substituted aromatic group, wherein each substituent of the aromatic group is independently halogen. In some embodiments, X 9 Comprising a side chain which is or comprises two optionally substituted aromatic groups. In some embodiments, X 9 Comprising a side chain which is or comprises an optionally substituted aromatic group, wherein each substituent of the aromatic group is independently selected from halogen or-OH. In some embodiments, the aromatic group is phenyl. In some embodiments, the aromatic group is an optionally substituted 8-to 10-membered bicyclic aryl or heteroaryl having 0 to 5 heteroatoms. In some embodiments, the aromatic group is an optionally substituted 9-to 10-membered bicyclic aryl or heteroaryl having one heteroatom. In some embodiments, X 9 Are residues of amino acids of the formulae A-I or salts thereof. In some embodiments, the amino acid residue has a formula of-NH-C (R a2 )(R a3 ) -a C (O) -structure or a salt thereof. In some embodiments, the amino acid residue has the formula-NH-CH (R a3 ) -C) O) -structure or salt thereof. R, as described herein a3 is-L a -R', wherein each variable is independently as described herein. In some embodiments, R' is R, as described herein. In some embodiments, R is an optionally substituted group selected from the group consisting of: phenyl, 10 membered bicyclic aryl, 5 to 6 membered heteroaryl having 1 to 4 heteroatoms and 9 to 10 membered bicyclic heteroaryl having 1 to 5 heteroatoms. In some embodiments, each substituent is independently halogen or-OH or C 1-6 Halogenated aliphatic. In some embodiments, each substituent is independently halogen or-OH. In some embodiments, R is optionally substituted phenyl. In some embodiments, R is phenyl. In some embodiments, R is optionally substituted aryl. In some embodiments, R is aryl. In some embodiments, R is an optionally substituted 5 membered heteroaryl having 1 to 4 heteroatoms. In some embodiments, R is an optionally substituted 5 membered heteroaryl having 1 heteroatom. In some embodiments, optionally substituted R is a 6 membered heteroaryl having 1 to 4 heteroatoms. In some embodiments, optionally substituted R is a 6 membered heteroaryl having 1 heteroatom. In some embodiments, R is an optionally substituted 9-membered heteroaryl having 1 to 5 heteroatoms. In some embodiments, R is an optionally substituted 9-membered heteroaryl having 1 heteroatom. In some embodiments, R is an optionally substituted 10 membered heteroaryl having 1 to 5 heteroatoms. In some embodiments, R is an optionally substituted 10 membered heteroaryl having 1 heteroatom. In some embodiments, the heteroatom is nitrogen. In some embodiments, the heteroatom is oxygen. In some embodiments, the heteroatom is sulfur. L as described herein a Is L. In some embodiments, L is a covalent bond. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution.
In some embodiments, X 9 Is a residue of an amino acid selected from the group consisting of: phe,3COOHF,2NapA, tyr,3Thi,4FF,4ClF,4BrF,3FF,3ClF,3BrF,2FF,3OMeF,4CNF,3CNF,4MeF,3MeF, aic, rbipPrF, sbipPrF, rbipPrDF, rbMeXyl A, rbMeXyIDA, bztA,1NapA, trp,2Thi,4TriA,3F3MeF, his, sbMeXyl A, and SbMeXyl DA.
In some embodiments, X 9 Is Phe. In some embodiments, X 9 Is 3COOHF. In some embodiments, X 9 Is 2NapA. In some embodiments, X 9 Is Tyr. In some embodiments, X 9 Is 3Thi. In some embodiments, X 9 Is 4FF. In some embodiments, X 9 Is 4C1F. In some embodiments, X 9 Is 4BrF. In some embodiments, X 9 Is 3FF. In some embodiments, X 9 Is 3ClF. In some embodiments, X 9 Is 3BrF. In some embodiments, X 9 Is 2FF. In some embodiments, X 9 Is 3ome f. In some embodiments, X 9 Is 4CNF. In some embodiments, X 9 Is 3CNF. In some embodiments, X 9 Is 4MeF. In some embodiments, X 9 Is 3MeF. In some embodiments, X 9 Is Aic. In some embodiments, X 9 Is RbiprF. In some embodiments, X 9 Is SbiprF. In some embodiments, X 9 Is RbiprDF. In some embodiments, X 9 Is RbMeXyl A. In some embodiments, X 9 Is RbMeXylDA. In some embodiments, X 9 Is BztA. In some embodiments, X 9 Is 1NapA. In some embodiments, X 9 Is Trp. In some embodiments, X 9 Is 2Thi. In some embodiments, X 9 Is 4TriA. In some embodiments, X 9 Is 3F3MeF. In some embodiments, X 9 Is His. In some embodiments, X 9 Is SbMeXy1A. In some embodiments, X 9 Is SbMeXy1DA.
In some embodiments, X 9 Is a residue of an amino acid whose side chain is hydrophobic. In some embodiments, X 9 Are hydrophobic amino acid residues as described herein. In some embodiments, X 9 Selected from nLeu, ala, cba, cypA, leu, ile, chg, val and 2Cpg.
In some embodiments, X 9 Is the residue of an amino acid comprising an acidic or polar group. In some embodiments, X 9 Is the residue of an amino acid whose side chain contains a polar group. In some embodiments, X 9 Are polar amino acid residues as described herein. In some embodiments, X 9 Is the residue of an amino acid whose side chain contains-OH. For example, in some embodiments, X 9 Is a residue of Ser. In some embodiments, X 9 Is a residue of Hse. In some embodiments, X 9 Is a compound whose side chain contains an amide group (e.g., -C (O) N (R') 2 For example-CONH 2 ) Residues of amino acids of (a). For example, in some embodiments, X 9 Is a residue of Asn. In some embodiments, X 9 Is Gln.
In some embodimentsIn the case of X 9 Phe, ala, lys,3COOHF, aib,2NapA, nLeu,2Thi, tyr,3Thi,4FF,4ClF,4BrF,3FF,3ClF,3BrF,2FF,3OMeF,4CNF,3CNF,4MeF,3MeF, aic, rbipPrF, sbipPrF, rbipPrDF, rbMeXyl A, rbMeXyl DA, cba, cypA, bztA, inapa, trp, leu, ile, ser, chg, hse,4TriA,3F3MeF, thr, his, val, asn, gln,2Cpg, sbMeXyl A, or SbMeXyl DA.
In some embodiments, X 9 Is Phe. In some embodiments, X 9 Is Ala.
In some embodiments, X 9 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-IV.
In some embodiments, X 9 Interacting with Lys345 of β -catenin or its corresponding amino acid residue. In some embodiments, X 9 Interact with Trp383 of beta-catenin or its corresponding amino acid residue. In some embodiments, X 9 Interact with Lys345 and Trp383 of β -catenin or their corresponding amino acid residues.
Various types of amino acid residues can be used for X 10 For example, amino acids of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or residues of salts thereof, according to the present disclosure. In some embodiments, X 10 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 10 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 10 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H.
In some embodiments, X 10 Is that
Lys, glnR, triAzLys, sla, dLys, asnR, hGlnR, iPrLys, triAzOrn, DGlnR, orn,4pipa, sch2s, [8fbb ] Cys, [ mXyl ] Cys, [ ohyl ] Cys, [ pXyl ] Cys, dsorn, dDab, NMeOrn, [2_b-naph ] Cys, or [3_3-biph ] Cys.
In some embodiments, X 10 Is Lys, glnR or TriAzLys. In some embodiments, X 10 Is Lys. In some embodiments, X 10 Is Gln. In some embodiments, X 10 Is TriAzLys. In some embodiments, X 10 Is sA. In some embodiments, X 10 Is dLys. In some embodiments, X 10 Is AsnR. In some embodiments, X 10 Is hGLnR. In some embodiments, X 10 Is iPrLys. In some embodiments, X 10 Is TriAzOrn. In some embodiments, X 10 Is DGlnR. In some embodiments, X 10 Is Orn. In some embodiments, X 10 Is 4PipA. In some embodiments, X 10 sCH2S. In some embodiments, X 10 Is [8FBB ]]Cys. In some embodiments, X 10 Is [4FB ]]Cys. In some embodiments, X 10 Is [ mXyl ]]Cys. In some embodiments, X 10 Is [ oXyl ]]Cys. In some embodiments, X 10 Is [ pXyl ]]Cys. In some embodiments, X 10 Is dOrn. In some embodiments, X 10 Is dDab. In some embodiments, X 10 Is NMeOrn. In some embodiments, X 10 Is [2_6-naph ]]Cys. In some embodiments, X 10 Is [3_3-biph ]]Cys。
In some embodiments, X 10 Are not stapled (e.g., when other residues are optionally stapled). In some embodiments, X 10 Is a residue of Leu or Phe. In some embodiments, X 10 Is a residue of Leu. In some embodiments, X 10 Is a residue of Phe.
In some embodiments, X 10 Are suitable amino acid residues for stapling, as described herein.
In some embodiments, the amino acid residue suitable for stapling is-N (R a1 )-L a1 -C(-L a -R SP1 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, it is-N (R a1 )-C(-L a -R SP1 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, each amino acid residue is independently-N (R a1 )-L a1 -C(-L a -R SP1 )(R a3 )-L a2 -C (O) -or-N (R) a1 )-C(-L a -R SP1 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H. In some embodiments, R a1 And R is a3 Both are-H.
In some embodiments, R is an amino acid residue in a pair SP1 is-NHR, wherein R is as described herein. In some embodiments, R is-H. In some embodiments, R is optionally substituted C 1-6 Aliphatic series. In some embodiments, R is optionally substituted C 1-6 An alkyl group. In some embodiments, R is C 1-6 Aliphatic series. In some embodiments, R is C 1-6 An alkyl group. In some embodiments, R SP1 is-NH 2 . In some embodiments, such amino acid residues may be stapled to another amino acid residue comprising-COOH by amidation to form a staple-like structure comprising-C (O) N (R') -, e.g., L s Wherein L is s2 is-C (O) N (R ') -or comprises-C (O) N (R') -. In some embodiments, L s2 is-C (O) N (R ') -wherein R' is as described herein. In some embodiments, R' is R, as described herein. In some embodiments, R is-H. In some embodiments, R is optionally substituted C 1-6 Aliphatic series. In some embodiments, R is optionally substituted C 1-6 An alkyl group. In some embodiments, R is methyl. In some embodiments, R is ethyl. In some embodiments, R is isopropyl. In some embodiments, -N (R'), -is fromAmino acid residues comprising an amino group prior to stapling. In some embodiments, -C (O) -is from an amino acid residue comprising-COOH or an activated form thereof prior to stapling. In some embodiments, R in another amino acid residue of a pair SP1 is-COOH or an active derivative thereof. In some embodiments, R in another amino acid residue of a pair SP1 is-COOH. In some embodiments, R' is R. In some embodiments, R' is-H. In some embodiments, L s1 Is a first amino acid residue (e.g., X 10 ) L of (2) a . In some embodiments, L s3 L being a second amino acid residue (e.g., the C-direction amino acid residue of the first amino acid residue) a . In some embodiments, the first amino acid residue is X 10 And the second amino acid residue is X 10 C-oriented amino acid residues of (A), e.g. X 14 . In some embodiments, L s1 And L s3 Each independently is L. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -、-C(O) -, -N (R') -, cy-or-O-. In some embodiments, L s1 And L s3 Each independently is L, wherein L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L s1 And L s3 Each independently is L, wherein L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L s -(CH 2 )n1-C(O)N(R’)-L s3 -wherein each variable is independently as described herein. In some embodiments, L s -L s1 -C(O)N(R’)-(CH 2 ) n 2-wherein each variable is independently as described herein. In some embodiments, L s -(CH 2 )n1-C(O)N(R’)-(CH 2 ) n 2-wherein each variable is independently as described herein. In some embodiments, n1 and n2 are each independently 1 to 10. In some embodiments, the first amino acid residue has R as an amino group SP1 And the second amino acid residue has R as-COOH or an activated form thereof SP1 . In some embodiments, the second amino acid residue has R as an amino group SP1 And the first amino acid residue has R as-COOH or an activated form thereof SP1 。
In some embodiments, the first amino acid residue is X 10 And the second amino acid residue is one of its C-direction amino acid residues, e.g., X 14 . In some embodiments, the second amino acid residue is X 10 And the first amino acid residue is one of its N-directional amino acid residues, e.g. X 7 。
In some embodiments, the first amino acid residue is X 10 . In some embodiments, X 10 Is Lys. In some embodiments, X 10 Is dLys. In some embodiments, X 10 Is iPrLys. In some embodiments, X 10 Is NMeOrn. In some embodiments, L s2 R 'of-N (R') -is optionally substituted C 1-6 An alkyl group. In some embodiments, it is methyl. In some embodiments, it is isopropyl. In some embodiments, n1 is 4. In some embodimentsIn which n1 is 3. In some embodiments, X 10 Is Orn. In some embodiments, X 10 Is dOrn. In some embodiments, n1 is 3. In some embodiments, X 10 Is dDab. In some embodiments, n1 is 2. In some embodiments, L s2 -N (R') -and L s1 And (5) bonding. In some embodiments, the second amino acid residue is X 14 . In some embodiments, X 14 Is GlnR. In some embodiments, X 14 Is hGLnR. In some embodiments, n1 is 4, as in Lys. In some embodiments, n2 is 2, as in GlnR. In some embodiments, n2 is 3.
In some embodiments, the first amino acid residue is X 10 It is 4PipA. In some embodiments, L s1 Is- (CH) 2 ) n1 -C(R’) 2 -(CH 2 ) n3 -wherein n1 and n3 are each independently n (e.g., 1 to 10,1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) as described herein, and each R' is independently as described herein. In some embodiments, one R' is-H. In some embodiments, n1 is 1. In some embodiments, n3 is 2. In some embodiments, - (CH) 2 ) n 3-and L s2 -N (R') -linkage. In some embodiments, L s1 Is (R') 2 -one R' and L s2 R 'of-N (R') -together with the intervening atoms thereof form an optionally substituted as described herein. In some embodiments, the ring formed is an optionally substituted 3-to 10-membered saturated ring. In some embodiments, the ring formed is 3-membered. In some embodiments, it is 4-membered. In some embodiments, it is 5-membered. In some embodiments, it is 6 membered. In some embodiments, it is 7-membered. In some embodiments, it is 8-membered. In some embodiments, the ring formed has no additional ring heteroatoms other than the nitrogen to which R' is attached. In some embodiments, L s is-L s1 -Cy-C(O)-L s3 -wherein each variable is independently as described herein. In some embodiments, -Cy-is optionalSubstitutedWherein the nitrogen atom is bonded to-C (O) -. In some embodiments, each L s1 And L s3 Independently is L, as described herein. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In some embodiments, L s Is- (CH) 2 )n1-Cy-C(O)-(CH 2 ) n2-, wherein each variable is independently as described herein. In some embodiments, -Cy-is optionally substituted +.>Wherein the nitrogen atom is bonded to-C (O) -. In some embodiments, n1 is 1. In some embodiments, the second amino acid residue is X 14 . In some embodiments, X 14 Is GlnR. In some embodiments, n2 is 2.
In some embodiments, the firstOne amino acid residue being X 7 Such as GlnR. In some embodiments, n1 is 2. In some embodiments, the second amino acid residue is X 10 Such as Lys. In some embodiments, n2 is 4. In some embodiments, the first amino acid residue is X 7 Such as Lys. In some embodiments, n1 is 4. In some embodiments, the second amino acid residue is X 10 Such as GlnR. In some embodiments, n2 is 2.
In some embodiments, the first amino acid residue is X 10 . In some embodiments, X 10 Is GlnR. In some embodiments, X 10 Is DGlnR. In some embodiments, n1 is 2. In some embodiments, X 10 Is AsnR. In some embodiments, n1 is 1. In some embodiments, L s2 C (O) -and L s1 And (5) bonding. In some embodiments, the first amino acid residue is X 10 Such as hGlnR. In some embodiments, n1 is 3. In some embodiments, the second amino acid residue is X 14 Such as iPrLys. In some embodiments, L s2 R 'of-N (R') -is optionally substituted C 1-6 An alkyl group. In some embodiments, it is isopropyl. In some embodiments, n2 is 4. In some embodiments, the second amino acid residue is X 14 Such as Lys. In some embodiments, the second amino acid residue is X 14 Such as Orn. In some embodiments, n2 is 3.
In some embodiments, the second amino acid residue is X 14 It is 4PipA. In some embodiments, L s3 Is- (CH) 2 ) n2 -C(R’) 2 -(CH 2 ) n3 -wherein n2 and n3 are each independently n (e.g., 1 to 10,1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) as described herein, and each R' is independently as described herein. In some embodiments, one R' is-H. In some embodiments, n2 is 1. In some embodiments, n3 is 2. In some embodiments, - (CH) 2 ) n 3-and L s2 -N (R') -linkage. In some embodimentsIn the case of L s3 Is (R') 2 -one R' and L s2 R 'of-N (R') -together with the intervening atoms thereof form an optionally substituted as described herein. In some embodiments, the ring formed is an optionally substituted 3-to 10-membered saturated ring. In some embodiments, the ring formed is 3-membered. In some embodiments, it is 4-membered. In some embodiments, it is 5-membered. In some embodiments, it is 6 membered. In some embodiments, it is 7-membered. In some embodiments, it is 8-membered. In some embodiments, the ring formed has no additional ring heteroatoms other than the nitrogen to which R' is attached. In some embodiments, L s is-L s1 -C(O)-Cy-L s3 -wherein each variable is independently as described herein. In some embodiments, -Cy-is optionally substitutedWherein the nitrogen atom is bonded to-C (O) -. In some embodiments, each L s1 And L s3 Independently is L, as described herein. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In some embodiments of the present invention, in some embodiments,l is an optionally substituted divalent straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In some embodiments, L s Is- (CH) 2 )n1-C(O)-Cy-(CH 2 ) n2-, wherein each variable is independently as described herein. In some embodiments, -Cy-is optionally substituted +.>Wherein the nitrogen atom is bonded to-C (O) -. In some embodiments, n1 is 2. In some embodiments, n2 is 1.
In some embodiments, the second amino acid residue is(e.g., X 14 ). In some embodiments, L s3 Is- (CH) 2 ) 2 -C(O)NH-(CH 2 ) 4 -. In some embodiments, the second amino acid residue is(e.g., X 14 ). In some embodiments, L s3 Is- (CH) 2 ) 2 -C (O) -Cy-. In some embodiments, -Cy-is optionally substituted +.>Wherein nitrogen is bonded to-C (O) -. In some embodiments, L s3 Is- (CH) 2 ) 2 -C(O)-N(R’)-(CH 2 ) n-CHR '-, in which two R's together with intervening atoms form an optionally substituted ring, as described herein. In some embodiments, the ring formed is optionally substituted +.>In some embodiments, the second amino acid residue is +.>(e.g., X 14 ). In some embodiments, L s3 Is- (CH) 2 ) 2 -C(O)-N(R’)-(CH 2 ) n-Cy-. In some embodiments, R' is R, as described herein. In some embodiments, R is-H. In some embodiments, R is optionally substituted C 1-6 Aliphatic series. In some embodiments, R is optionally substituted C 1-6 An alkyl group. In some embodiments, R is methyl. In some embodiments, n is 1. In some embodiments, -Cy-is optionally substituted +.>Wherein nitrogen is with L s2 Bonding, L s2 is-C (O) -or comprises-C (O) -. In some embodiments, L s3 Is- (CH) 2 ) 2 -C(O)-N(R’)-CH 2 -CHR’-(CH 2 ) n-. In some embodiments, n is 2. In some embodiments, - (CH) 2 ) n-and L s2 -N (R') -linkage, L s2 is-C (O) -N (R') -. In some embodiments, L s3 R 'and L of-CHR' -, are described s2 R 'of-N (R') -and intervening atoms thereof taken together form an optionally substituted ring, as described herein. In some embodiments, the ring formed is optionally substituted +.>In some embodiments, the second amino acid residue is +.>(e.g., X 14 ). In some embodiments, the second amino acid residue is(e.g., X 14 ). In some embodiments L s3 Is- (CH) 2 ) 2 -C(O)-N(R’)-(CH 2 ) n1 -C(R’) 2 -(CH 2 ) n2 -. In some embodimentsN1 and n2 are each independently 1 to 10. In some embodiments, n1 is 1. In some embodiments, n1 is 2. In some embodiments, n2 is 2. In some embodiments, -N (R '), -R ' and-C (R ') 2 -one R' taken together with the intervening atoms thereof forms an optionally substituted ring, as described herein. In some embodiments, the ring formed is an optionally substituted 6 membered monocyclic saturated ring, which ring has no heteroatoms other than the nitrogen atom of-N (R') -. In some embodiments, L s2 is-C (O) N (R') -. In some embodiments, -N (R'), -and- (CH) 2 ) n2 -bonding. In some embodiments, L s3 Is (R') 2 -one R' and L s2 R 'of-N (R') -and intervening atoms thereof taken together form an optionally substituted ring, as described herein. In some embodiments, the ring formed is an optionally substituted 6 membered monocyclic saturated ring, which ring has no heteroatoms other than the nitrogen atom of-N (R') -.
In some embodiments, the first amino acid residue is(e.g., X 10 ). In some embodiments, L s1 Is- (CH) 2 ) 2 -C(O)-N(R’)-(CH 2 ) n-CHR '-, in which two R's together with intervening atoms form an optionally substituted ring, as described herein. In some embodiments, the ring formed is optionally substitutedIn some embodiments, the second amino acid residue is GlnR (e.g., X 14 ). In some embodiments, L s3 Is- (CH) 2 ) 2 -。
In some embodiments, the first amino acid residue is(e.g., X 10 ). In some embodiments, L s1 Is- (CH) 2 ) 2 -C(O)-N(R’)-(CH 2 ) n-Cy-. In some embodiments, R' is R, as described herein. In some embodiments, R is-H. In some embodiments, R is optionally substituted C 1-6 Aliphatic series. In some embodiments, R is optionally substituted C 1-6 An alkyl group. In some embodiments, R is methyl. In some embodiments, n is 1. In some embodiments, -Cy-is optionally substituted +.>Wherein nitrogen is with L s2 Bonding, L s2 is-C (O) -or comprises-C (O) -. In some embodiments, L s1 Is- (CH) 2 ) 2 -C(O)-N(R’)-CH 2 -CHR’-(CH 2 ) n-. In some embodiments, n is 2. In some embodiments, - (CH) 2 ) n-and L s2 -N (R') -linkage, L s2 is-C (O) -N (R') -. In some embodiments, L s1 R 'and L of-CHR' -, are described s2 R 'of-N (R') -and intervening atoms thereof taken together form an optionally substituted ring, as described herein. In some embodiments, the ring formed is optionally substituted +.>In some embodiments, the first amino acid residue is +.>(e.g., X 10 ). In some embodiments, the first amino acid residue is +.>(e.g., X 10 )。
In some embodiments, L s1 Is- (CH) 2 ) 2 -C(O)-N(R’)-(CH 2 ) n1 -C(R’) 2 -(CH 2 ) n2 -. In some embodiments, n1 and n2 are each independently 1 to 10. In some embodiments, n1 is 1. In some embodimentsIn which n1 is 2. In some embodiments, n2 is 2. In some embodiments, -N (R '), -R ' and-C (R ') 2 -one R' taken together with the intervening atoms thereof forms an optionally substituted ring, as described herein. In some embodiments, the ring formed is an optionally substituted 6 membered monocyclic saturated ring, which ring has no heteroatoms other than the nitrogen atom of-N (R') -. In some embodiments, L s2 is-C (O) N (R') -. In some embodiments, -N (R'), -and- (CH) 2 ) n2 -bonding. In some embodiments, L s1 Is (R') 2 -one R' and L s2 R 'of-N (R') -and intervening atoms thereof taken together form an optionally substituted ring, as described herein. In some embodiments, the ring formed is an optionally substituted 6 membered monocyclic saturated ring, which ring has no heteroatoms other than the nitrogen atom of-N (R') -. In some embodiments, the second amino acid residue is GlnR (e.g., X 14 )。
In some embodiments, the first residue is(e.g., X 10 ). In some embodiments, the first residue is +. >(e.g., X 10 ). In some embodiments, the first residue is +.>(e.g., X 10 ). In some embodiments, L s1 Is- (CH) 2 ) N-N (R') -C (O) -Cy-Cy-, wherein each variable is independently as described herein. In some embodiments, L s1 Is- (CH) 2 ) N-N (R') -C (O) -Cy-, wherein each variable is independently as described herein. In some embodiments, the first residue is +.>(e.g., X 10 ). In one placeIn some embodiments, L s1 Is- (CH) 2 )n-N(R’)-C(O)-CH 2 -, wherein R is as described herein and is bonded to C (O) -, a-CH 2 -is optionally substituted. In some embodiments, L s1 Is- (CH) 2 )n-N(R’)-C(O)-C(R’) 2 -, wherein each R is independently as described herein. In some embodiments, L s1 Is- (CH) 2 )n-N(R’)-C(O)-C(CH 3 ) 2 -, wherein R is as described herein. In some embodiments, the first residue is +.>(e.g., X 10 ). In some embodiments, L s1 Is- (CH) 2 ) n1 -N(R’)-C(O)-(CH 2 ) n2 -wherein each variable is independently as described herein. In some embodiments, n1 and n2 are each independently n as described herein. In some embodiments, L s1 Is- (CH) 2 ) 4 -N(R’)-C(O)-(CH 2 ) 2 -, wherein each R is independently as described herein. In some embodiments, n is 1 to 10. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, R' is R, as described herein. In some embodiments, R is-H. In some embodiments, -Cy-is optionally substituted phenylene. In some embodiments, -Cy-is optionally substituted 1, 2-phenylene. In some embodiments, -Cy-is optionally substituted 1, 3-phenylene. In some embodiments, each-Cy-is independently optionally substituted 1, 2-phenylene. In some embodiments, each-Cy-is independently optionally substituted 1, 3-phenylene. In some embodiments, L s2 is-C (O) -N (R ') -or comprising-C (O) -N (R') -, as described herein. In some embodiments, R' is R, as described herein. In some embodiments, R is-H. In some embodiments, L s2 is-C (O) NH-. In some embodiments, -C (O) -and L s1 -Cy-bond of (c). In some implementationsIn embodiments, the second residue is X 14 For example, lys. In some embodiments, L s3 Is, for example, optionally substituted- (CH) as described herein 2 ) n-. In some embodiments, L s3 Is- (CH) 2 ) n-. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4 (e.g., as in Lys).
In some embodiments, R is an amino acid residue in a pair SP1 Is the first reactive group of the cycloaddition reaction. In some embodiments, such an amino acid residue may be stapled by a cycloaddition reaction with another amino acid residue comprising a second reactive group of the cycloaddition reaction. In some embodiments, R in another amino acid residue of a pair SP1 Is the second reactive group of the cycloaddition reaction. In some embodiments, the cycloaddition reaction is [3+2 ] ]. In some embodiments, the cycloaddition reaction is a click chemistry reaction. In some embodiments, the cycloaddition reaction is [4+2 ]]. In some embodiments, one of the first and second reactive groups is-N 3 Or comprises-N 3 And the other is or comprises an alkyne (e.g., a terminal alkyne or an activated/strained alkyne).
In some embodiments, R of the first amino acid residue SP1 is-N 3 . In some embodiments, L of the first amino acid residue a Is L, as described herein. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodimentsWherein L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution.
In some embodiments, R of the second amino acid residue SP1 is-C.ident.C-or contains-C.ident.C-. In some embodiments, R of the second amino acid residue SP1 Is thatIn some embodiments, R of the second amino acid residue SP1 Including, for example, strained alkynes in the ring. In some embodiments, L of the first amino acid residue a Is L, as described herein. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In some embodiments, L is optionally substituted diValence straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution.
In some embodiments, L s is-L s1 -L s2 -L s3 -, wherein L s2 is-Cy-or comprises-Cy-. In some embodiments, L s2 is-Cy-. In some embodiments, -Cy-is formed by a cycloaddition reaction. In some embodiments, -Cy-is optionally substitutedIn some embodiments, -Cy-is +.>In some embodiments, -Cy-is optionally substituted +.>In some embodiments, -Cy-is +.>In some embodiments, L s1 L being the first amino acid residue a And L is s3 L being a second amino acid residue a . In some embodiments, L s1 L being a second amino acid residue a And L is s3 L being the first amino acid residue a . In some embodiments, L s1 And L s3 Each independently is L, as described herein. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is- (CH) 2 ) n-, wherein n is 1 to 10. In some implementationsIn embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In some embodiments, L s1 Is optionally substituted- (CH) 2 ) n -, wherein n is 1 to 10. In some embodiments, L s1 Is- (CH) 2 ) n -, wherein n is 1 to 10. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, L s3 Is optionally substituted- (CH) 2 ) n -, wherein n is 1 to 10. In some embodiments, L s3 Is- (CH) 2 ) n -, wherein n is 1 to 10. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4.
In some embodiments, the first amino acid residue is X 10 . In some embodiments, X 10 R of (2) SP1 is-N 3 . In some embodiments, X 10 L of (2) a Is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, X 10 L of (2) a Is- (CH) 2 ) 4 -. In some embodiments, X 10 L of (2) a Is- (CH) 2 ) 3 -. In some embodiments, X 10 L of (2) a Is- (CH) 2 ) 2 -. In some embodiments, X 10 L of (2) a is-CH 2 -. In some embodiments, the second amino groupThe acid residue being X 14 . In some embodiments, X 14 R of (2) SP1 Is or comprises an alkyne, for example, a strained/activated alkyne. In some embodiments, X 14 R of (2) SP1 is-C.ident.CH. In some embodiments, X 14 L of (2) a Is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, X 14 L of (2) a Is- (CH) 2 ) 4 -. In some embodiments, X 14 L of (2) a Is- (CH) 2 ) 3 -. In some embodiments, X 14 L of (2) a Is- (CH) 2 ) 2 -. In some embodiments, X 14 L of (2) a is-CH 2 -. In some embodiments, the methylene units are replaced with-O-. In some embodiments, X 14 L of (2) a is-CH 2 -O-CH 2 -. In some embodiments, L s3 Is X 14 L of (2) a . In some embodiments, L s3 And L is equal to s2 Is bonded to a carbon atom of (c).
In some embodiments, the first amino acid residue is X 10 . In some embodiments, X 10 R of (2) SP1 Is or comprises an alkyne, for example, a strained/activated alkyne. In some embodiments, X 10 R of (2) SP1 is-C.ident.CH. In some embodiments, X 10 L of (2) a Is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, X 10 L of (2) a Is- (CH) 2 ) 4 -. In some embodiments, X 10 L of (2) a Is- (CH) 2 ) 3 -. In some embodiments, X 10 L of (2) a Is- (CH) 2 ) 2 -. In some embodiments, X 10 L of (2) a is-CH 2 -. In some embodiments, the methylene units are replaced with-O-. In some embodiments, X 10 L of (2) a is-CH 2 -O-CH 2 -. In some embodiments, L s1 Is X 10 L of (2) a . In some embodiments, L s1 And L is equal to s2 Is bonded to a carbon atom of (c). In some embodiments, the second amino acid residue is X 14 . In some embodiments, X 14 R of (2) SP1 is-N 3 . In some embodiments, X 14 L of (2) a Is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, X 14 L of (2) a Is- (CH) 2 ) 4 -. In some embodiments, X 14 L of (2) a Is- (CH) 2 ) 3 -. In some embodiments, X 14 L of (2) a Is- (CH) 2 ) 2 -. In some embodiments, X 14 L of (2) a is-CH 2 -。
In some embodiments, R SP1 Is a nucleophile. In some embodiments, R SP1 is-SH, e.g., as in Cys. In some embodiments, L s2 Is L ", as described herein. In some embodiments, L s2 is-S-CH 2 -L”-CH 2 -S-, wherein L "is as described herein. In some embodiments, the staple-like structure has-L s1 -S-CH 2 -L”-CH 2 -S-L s3 -structure, wherein each variable is independently as described herein, and each-CH 2 -is optionally substituted. In some embodiments, L s2 is-S-C (R') 2 -L”-C(R’) 2 -S-, wherein each variable is independently as described herein. In some embodiments, the staple-like structure has-L s1 -S-C(R’) 2 -L”-C(R’) 2 -S-L s3 -structure wherein each variable is independently as described herein. In some embodiments, each R' is independently R, as described herein. In some embodiments, each R' is-H. In some embodiments, L s2 is-S-Cy-S-, wherein-Cy-is as described herein. In some embodiments, the staple-like structure has-L s1 -S-Cy-S-L s3 -structure wherein each variable is independently as described herein. In some embodiments, L s2 is-S-Cy-Cy-S-, wherein-Cy-is as described herein. In some embodiments, the staplerThe staple-like structure has-L s1 -S-Cy-Cy-S-L s3 -structure wherein each variable is independently as described herein. In some embodiments, L s1 L being the first amino acid residue a . In some embodiments, L s3 L being a second amino acid residue a . In some embodiments, L s1 And L s3 Each is L, as described herein. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In some embodiments, each of the pair of amino acid residues is Cys. In some embodiments, L s1 is-CH 2 -. In some embodiments, L s3 is-CH 2 -. In some embodiments, L "is-Cy-, as described herein. In some embodiments, -Cy-is optionally substituted phenylene. In some embodiments, -Cy-is phenylene. In some embodiments, -Cy-is optionally substituted 1, 2-phenylene. In some embodiments, -Cy-is 1, 2-phenylene. In some embodiments, -Cy-isOptionally substituted 1, 3-phenylene. In some embodiments, -Cy-is 1, 3-phenylene. In some embodiments, -Cy-is optionally substituted 1, 4-phenylene. In some embodiments, -Cy-is tetrafluoro-1, 4-phenylene. In some embodiments, -Cy-is 1, 4-phenylene. In some embodiments, -Cy-is optionally substituted naphthylene. In some embodiments, -Cy-is optionally substituted In some embodiments, L "is-Cy-, wherein each-Cy-is independently as described herein. In some embodiments, each-Cy-is independently optionally substituted phenylene. In some embodiments, each-Cy-is independently phenylene. In some embodiments, each-Cy-is independently optionally substituted 1, 2-phenylene. In some embodiments, each-Cy-is independently 1, 2-phenylene. In some embodiments, each-Cy-is independently optionally substituted 1, 3-phenylene. In some embodiments, each-Cy-is independently 1, 3-phenylene. In some embodiments, each-Cy-is independently optionally substituted 1, 4-phenylene. In some embodiments, each-Cy-is independently 1, 4-phenylene. In some embodiments, each-Cy-is independently tetrafluoro-1, 4-phenylene.
As will be appreciated by those skilled in the art, such staple-like structures may be formed by using a staple-like structure having R x -L s2 -R x The linker reagent of the structure is formed by linking Cys residues, wherein each variable is independently as described herein. In some embodiments, each R x Are all-Br.
In some embodiments, R of two amino acid residues of a pair of amino acid residues suitable for stapling SP1 Can each independently react with a linking agent to form a staple-like structure. In some embodiments, suitable linking reagents comprise two reactive groups, each of which may be independently associated with R of each amino acid residue SP1 And (3) reacting. In some embodiments, the linking agent has an H-L "-H structure or salt thereof, wherein the agent packageContaining two amino groups and L' is a covalent bond or an optionally substituted divalent C 1- C 20 An aliphatic group wherein one or more methylene units of the aliphatic group are optionally and independently substituted
-C(R’) 2 -，-Cy-,-O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -,-S(O) 2 N (R') -C (O) S-, or-C (O) O-. In some embodiments, such linking reagents can react with two amino acid residues, each independently having R SP1 A group which is-COOH or an activated form thereof.
Some suitable embodiments of L "include those described herein for L that fall within the scope of L". For example, in some embodiments, L "is L, wherein L is an optionally substituted divalent straight or branched chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution.
In some embodiments, the linking agent is a diamine or salt thereof. In one placeIn some embodiments, the agent has a NHR-L "-NHR structure or a salt thereof, wherein each variable is independently as described herein. In some embodiments, each R is independently-H or optionally substituted C 1-6 Aliphatic series. In some embodiments, each R is independently-H or C 1-6 Aliphatic series. In some embodiments, each R is independently-H or optionally substituted C 1-6 An alkyl group. In some embodiments, each R is independently-H or C 1-6 An alkyl group. In some embodiments, the reagent has NH 2 -L”-NH 2 A structure or a salt thereof. In some embodiments, L "is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L "is- (CH) 2 ) 4 -。
In some embodiments, staple-like structure L s is-L s1 -L s2 -L s3 -, wherein L s1 L being the first amino acid residue of a stapled pair a ，L s3 L being the second amino acid residue of a stapled pair a And L is s2 is-C (O) -N (R ') -L "-N (R') -C (O) -, wherein each variable is independently as described herein. In some embodiments, L "is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L "is- (CH) 2 ) 4 -. In some embodiments, L s1 And L s3 Each independently is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, n is 2. In some embodiments, the first amino acid residue is Gln (e.g., X 10 ). In some embodiments, the second amino acid residue is GlnR (e.g., X 14 ). In some embodiments, two GlnRs can be obtained by [ diaminobutane]Such a staple-like structure is formed.
In some embodiments, the linking reagent has an H-Cy-L "-NHR structure or salt thereof, wherein-Cy-comprises a second amino group. In some embodiments, R is-H or optionally substituted C 1-6 Aliphatic series. In some embodiments, R is-H or C 1-6 Aliphatic series. In some embodiments, R is-H or optionally substituted C 1-6 An alkyl group. In some embodiments, R is-H or C 1-6 An alkyl group. In some embodiments, R is methyl. In some embodiments, the linking reagent has H-Cy-L "-NH 2 A structure or a salt thereof, wherein-Cy-comprises a second amino group. In some embodiments, -Cy-is optionally substitutedIn some embodiments, -Cy-is +.>In some embodiments, L "is a covalent bond. In some embodiments, L "is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L "is- (CH) 2 ) -. In some embodiments, the linking agent is +.>Or a salt thereof. In some embodiments, the linking agent isOr a salt thereof.
In some embodiments, L s2 is-C (O) -Cy-N (R') -C (O) -, wherein each variable is independently as described herein. In some embodiments, R' is-H. In some embodiments, -Cy-isIn some embodiments, L s1 And L s3 Each independently is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, n is 2. In some embodiments, -Cy-is closer to the N-terminus than-N (R') -. In some embodiments, -Cy-is closer to the C-terminus than-N (R') -. In some embodiments, the first amino acid residue is Gln (e.g., X 10 ). In some embodiments, the second amino acid residue is GlnR (e.g., X 14 ). In some embodiments, two GlnRs can be substituted by [4 aminopiperidines]Formation ofSuch a staple-like structure.
In some embodiments, L s2 is-C (O) -Cy- (CH) 2 ) N-N (R') -C (O) -, wherein each variable is independently as described herein. In some embodiments, R' is-H. In some embodiments, R' is R, as described herein, e.g., optionally substituted C 1-6 Aliphatic, C 1-6 Alkyl groups, and the like. In some embodiments, R is methyl. In some embodiments, n is 1. In some embodiments, -Cy-isIn some embodiments, -Cy-is closer to the N-terminus than-N (R') -. In some embodiments, -Cy-is closer to the C-terminus than-N (R') -. In some embodiments, L s1 And L s3 Each independently is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, n is 2. In some embodiments, the first amino acid residue is Gln (e.g., X 10 ). In some embodiments, the second amino acid residue is GlnR (e.g., X 14 ). In some embodiments, two GlnRs can be obtained by [4 mamplatidine ]]Such a staple-like structure is formed.
In some embodiments, the methylene units are replaced with-Cy-. In some embodiments, the linking reagent has an H-Cy-H structure, wherein Cy comprises two secondary amino groups. In some embodiments, -Cy-is an optionally substituted 8-to 20-membered bicyclic ring. In some embodiments, H-Cy-H comprises two-NH-. In some embodiments, -Cy-is optionally substitutedIn some embodiments, -Cy-is optionally substituted +.>In some embodiments, the meta-position linkage site (relative to the spiro carbon atom) is closer to the N-terminus than the para-position linkage site (relative to the spiro carbon atom). In some embodiments, the meta-position linkage site (relative to the spiroThe ring carbon atom) is closer to the C-terminus than the para-linkage site (relative to the spiro carbon atom).
In some embodiments, L s2 is-C (O) -Cy-C (O) -, wherein-Cy-is as described herein. In some embodiments, L s1 And L s3 Each independently is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, n is 2. In some embodiments, the first amino acid residue is Gln (e.g., X 10 ). In some embodiments, the second amino acid residue is GlnR (e.g., X 14 ). In some embodiments, two GlnRs can be obtained by reacting [29N2 spiroundecane ]Such a staple-like structure is formed. In some embodiments, two GlnRs can be obtained by reacting [39N2 spiroundecane]Such a staple-like structure is formed.
In some embodiments, the pairs of amino acid residues suitable for stapling both independently have-N (R a1 )-L a1 -C(-L a -R SP1 )(R a3 )-L a2 -C (O) -or-N (R) a1 )-C(-L a -R SP1 )(R a3 ) -C (O) -structure, wherein each variable is independently as described herein, and R SP1 Is an amino group. In some embodiments, R SP1 is-NHR, wherein R is as described herein. In some embodiments, R is-H. In some embodiments, R is optionally substituted C 1-6 Aliphatic series. In some embodiments, R is optionally substituted C 1-6 An alkyl group. In some embodiments, R is C 1-6 Aliphatic series. In some embodiments, R is C 1-6 An alkyl group. In some embodiments, R SP1 is-NH 2 . In some embodiments, such two amino acid residues may be linked by a diacid linking reagent.
In some embodiments, the linking agent has the HOOC-L "-COOH structure or a salt or activated form thereof, wherein L" is as described herein. In some embodiments, L "is-Cy-. In some embodiments, L "is-Cy-. In some embodiments, -Cy-is optionally substituted phenylene. In some embodiments, -Cy-is optionally substituted 1, 2-phenylene . In some embodiments, -Cy-is optionally substituted 1, 3-phenylene. In some embodiments, each-Cy-is independently optionally substituted 1, 2-phenylene. In some embodiments, each-Cy-is independently optionally substituted 1, 3-phenylene. In some embodiments, L "is optionally substitutedIn some embodiments, the linking agent is +.>Or a salt or activated form thereof. In some embodiments, L "is optionally substituted +.>In some embodiments, the linking agent is +.>Or a salt or activated form thereof. In some embodiments, L' is 1, 3-phenylene. In some embodiments, the linking agent is +.>Or a salt or activated form thereof. In some embodiments, L "is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L' is an optionally substituted-CH 2 -. In some embodiments, L 'is-C (R') 2 -. In some embodiments, L' is-C (CH 3 ) 2 -. In some embodiments, the linking agent is (CH 3 ) 2 C(COOH) 2 Or a salt or activated form thereof. In some embodiments, L' is-CH 2 CH 2 -. In some embodiments, the linking agent is hoogch 2 CH 2 COOH or a salt or activated form thereof.
In some embodiments, the staple-like structure is L s Wherein L is s2 is-N (R ') -L ' -N (R ') -and L s1 And L s3 Each independently as described herein. In some embodiments, L "is-Cy-, wherein each-Cy-is independently as described herein. In some embodiments, L "is-Cy-, as described herein. In some embodiments, -Cy-is optionally substituted phenylene. In some embodiments, -Cy-is optionally substituted 1, 2-phenylene. In some embodiments, -Cy-is optionally substituted 1, 3-phenylene. In some embodiments, each-Cy-is independently optionally substituted 1, 2-phenylene. In some embodiments, each-Cy-is independently optionally substituted 1, 3-phenylene. In some embodiments, L "is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L' is an optionally substituted-CH 2 -. In some embodiments, L 'is-C (R') 2 -. In some embodiments, L' is-C (CH 3 ) 2 -. In some embodiments, L' is-CH 2 CH 2 -. In some embodiments, L s1 And L s3 Each independently is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, n is 2. In some embodiments, n is 4. In some embodiments, the first amino acid residue is Lys (e.g., X 10 ). In some embodiments, the second amino acid residue is Lys (e.g., X 14 ). In some embodiments, two lyss may pass through [ biphenyl 33COOH]Such a staple-like structure is formed. In some embodiments, two lyss may be linked by [ dibenzoate]Such a staple-like structure is formed. In some embodiments, two lyss may be prepared by [ isophthalate]Such a staple-like structure is formed. In some embodiments, two lyss may be linked by [ Me2Mal]Such a staple-like structure is formed. In some embodiments, two Lys may be performed by [ succinate]Such a staple-like structure is formed.
In some embodiments, X 10 Is stapled. In some embodiments, X 10 And X is 14 Stapling. In some embodiments, X 10 And X is 7 Stapling.
In some embodimentsIn the scheme, X 10 Is Lys, phe, triAzLys, glnR, leu, pyrS2, aib, ala, sAla, asnR, hGLnR, dOrn, pyrS1, dLys, dDab, [ mPyr ]]Cys,PyrS3，iPrLys,[mXyl]Cys，TriAzOrn，1MeK，[C3]Cys，[IsoE]Cys,DGlnR,Orn,[mPyr]hCys,[Red]Cys,[C3]hCys,4PipA,sCH2S，[8FBB]Cys，[pXyl]Cys，[pXyl]hCys，[33Oxe]Cys，[Red]hCys，[IsoE]hCys,[13Ac]hCys,[m5Meb]Cys，[m5Meb]hCys,GlnS3APyr，AsnMeEDA，AsnR3APyr，[m5Pyr]Cys，[m5OMeb]Cys，[4FB]Cys，[oXyl]Cys，NMeOrn，[2_6-naph]Cys，[3_3-biph]Cys，[mXyl]hCys，[3_3-biPh]hCys，[2_6-naph]hCys，[33Oxe]hCys，[13Ac]Cys，GlnR3APyr，AsnS3APyr，[IsoE]hCysOx, or [ m5Pyr]hCys。
In some embodiments, X 10 Is Lys. In some embodiments, X 10 Is Phe. In some embodiments, X 10 Is TriAxLys. In some embodiments, X 10 Is GlnR. In some embodiments, X 10 Is Leu. In some embodiments, X 10 Is PryS2. In some embodiments, X 10 Is Aib. In some embodiments, X 10 Is Ala. In some embodiments, X 10 Is Val.
In some embodiments, X 10 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-I, tables A-II, tables A-III and tables A-IV.
Various types of amino acid residues can be used for X 11 For example, amino acids of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or residues of salts thereof, according to the present disclosure. In some embodiments, X 11 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 11 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 11 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodiments of the present invention, in some embodiments,R a3 is-H.
In some embodiments, X 11 Are residues of amino acids suitable for stapling, as described herein. In some embodiments, the amino acid residue suitable for stapling is-N (R a1 )-L a1 -C(-L a -R SP1 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, it is-N (R a1 )-C(-L a -R SP1 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, each amino acid residue is independently-N (R a1 )-L a1 -C(-L a -R SP1 )(R a3 )-L a2 -C (O) -or-N (R) a1 )-C(-L a -R SP1 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H. In some embodiments, R a1 And R is a3 Both are-H. In some embodiments, R SP1 Comprising optionally substituted-ch=ch-. In some embodiments, R SP1 Is optionally substituted-ch=ch 2 Or comprises optionally substituted-ch=ch 2 . In some embodiments, R SP1 is-CH=CH 2 。
In some embodiments, X 11 Are residues of amino acids, for example, having the structures of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., whose side chains contain functional groups suitable for stapling, such as double bonds. In some embodiments, X 11 Is a residue of an amino acid comprising one and no more than one functional group for stapling. In some embodiments, X 11 Is a residue of an amino acid comprising one and no more than one double bond for stapling. As in X 1 In some embodiments, X 11 Contains a ring structure and the amino group of which is part of the ring. In some embodiments, X 11 Is as followsAmino acids described herein (e.g., amino acids of formulas A-I, A-II, A-III, etc.), wherein R a1 And R is a3 Taken together form an optionally substituted ring, e.g., an optionally substituted 3-to 10-membered ring. In some embodiments, R a1 And R is a3 Taken together with intervening atoms, form an optionally substituted 3-to 10-membered saturated or partially saturated ring having 0 to 5 heteroatoms in addition to intervening atoms.
In some embodiments, R a2 And R is a3 Taken together form an optionally substituted ring, e.g., an optionally substituted 3-to 10-membered ring. In some embodiments, R a2 And R is a3 Taken together with intervening atoms, form an optionally substituted 3-to 10-membered saturated or partially saturated ring having 0 to 5 heteroatoms in addition to intervening atoms.
In some embodiments, as described herein, for example, at X 11 Or another part, by R a1 And R is a3 Together with intervening atoms, through R a2 And R is a3 The ring formed with the intervening atoms or with the intervening atoms by any other two suitable R's is saturated. In some embodiments, the ring formed is monocyclic. In some embodiments, the formed ring has no heteroatoms other than intervening atoms. In some embodiments, the formed ring has at least one heteroatom in addition to the intervening atoms. In some embodiments, the formed ring has at least one nitrogen in addition to the intervening atoms. In some embodiments, L a1 And L a2 Is a covalent bond. In some embodiments, the ring formed is unsubstituted. In some embodiments, the ring formed is substituted. In some embodiments, the substituent comprises a double bond that is suitable for metathesis with another double bond to form a staple-like structure. In some embodiments, the substituents have the formula-C (O) -O- (CH) 2 )n-CH＝CH 2 Structure wherein n is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10. In some embodiments, the substituent is bonded to a nitrogen ring atom (see, e.g., pyrS 2). In some embodiments, X 11 Is PyrS 2.
In some embodiments, L a Is- (CH) 2 ) n1 -N(R’)-C(O)-(CH 2 ) n2 -, wherein each variable is independently as described herein, and each-CH 2 -is optionally substituted. In some embodiments, L a Is- (CH) 2 ) n1 -N(R’)-C(O)-(CH 2 ) n2 -wherein each variable is independently as described herein. In some embodiments, - (CH) 2 ) n1 -and X 11 And (5) bonding. In some embodiments, n1 is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, n1 is 1. In some embodiments, n1 is 2. In some embodiments, n1 is 3. In some embodiments, n2 is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, n2 is 1. In some embodiments, n2 is 2. In some embodiments, n2 is 3. In some embodiments, n2 is 4. In some embodiments, n2 is 5. In some embodiments, R a3 And L a R 'in-N (R') -together with the intervening atoms form an optionally substituted ring. In some embodiments, the ring formed is an optionally substituted 3-to 10-membered monocyclic, saturated or partially unsaturated ring having 0 to 3 heteroatoms in addition to the nitrogen atom to which R' is attached. In some embodiments, the formed ring is saturated. In some embodiments, the ring formed is 3-membered. In some embodiments, the ring formed is 4-membered. In some embodiments, the ring formed is 5-membered. In some embodiments, the ring formed is 6 membered. In some embodiments, the ring formed is 7 membered. In some embodiments, the ring formed is 8-membered. In some embodiments, the formed ring has no ring heteroatoms other than the nitrogen atom to which R' is attached. In some embodiments, X 11 Is a residue of pyrS 2.
In some embodiments, X 11 Is stapled. In some embodiments, X 11 And X is 4 Stapling. In some embodiments, X 11 Is PyrS2 and stapled. In one placeIn some embodiments, X 11 Is Lys and stapled.
In some embodiments, X 11 Is a residue of pyrS2 or Lys.
In some embodiments, X 11 Is a residue of PyrS2 and is stapled.
In some embodiments, staple-like structures, e.g., L s Has a structure of-L s1 -L s2 -L s3 Structure, wherein each variable is independently as described herein. In some embodiments, L s1 Or L s3 Is X 11 L of (2) a As described herein. In some embodiments, L s3 Is X 11 L of (2) a As described herein. In some embodiments, L s1 Is another amino acid residue such as X 4 L of (2) a . In some embodiments, L s1 Is L, as described herein. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In one placeIn some embodiments, L s3 Is L, as described herein. In some embodiments, L s3 Is- (CH) 2 ) n1 -N(R’)-C(O)-(CH 2 ) n2 -, wherein each variable is independently as described herein, and each-CH 2 -is optionally substituted. In some embodiments, L s3 Is- (CH) 2 ) n1 -N(R’)-C(O)-(CH 2 ) n2 -wherein each variable is independently as described herein. In some embodiments, - (CH) 2 ) n1 -and X 11 And (5) bonding. In some embodiments, n1 is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, n1 is 1. In some embodiments, n1 is 2. In some embodiments, n1 is 3. In some embodiments, n2 is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, n2 is 1. In some embodiments, n2 is 2. In some embodiments, n2 is 3. In some embodiments, n2 is 4. In some embodiments, n2 is 5. In some embodiments, R a3 And L a R 'in-N (R') -together with the intervening atoms form an optionally substituted ring. In some embodiments, the ring formed is an optionally substituted 3-to 10-membered monocyclic, saturated or partially unsaturated ring having 0 to 3 heteroatoms in addition to the nitrogen atom to which R' is attached. In some embodiments, the formed ring is saturated. In some embodiments, the ring formed is 3-membered. In some embodiments, the ring formed is 4-membered. In some embodiments, the ring formed is 5-membered. In some embodiments, the ring formed is 6 membered. In some embodiments, the ring formed is 7 membered. In some embodiments, the ring formed is 8-membered. In some embodiments, the formed ring has no ring heteroatoms other than the nitrogen atom to which R' is attached.
In some embodiments, L s2 Is optionally substituted-ch=ch-. In some embodiments, L s2 Is-ch=ch-. In some embodiments, L s2 Is optionally substituted-CH 2 -CH 2 -. In some embodiments, L s2 is-CH 2 -CH 2 -。
In some embodiments, X 11 Is that
PyrS2, lys,3thi, ala, phe, SPIP3, pyrSadNap 3 butene, SPIP2, az3, dapAc7EDA, leu,3 allyloxy PyrSa, pyrSaV butene, az2, pyrS1, pyrSc72SMe3ROMe, pyrSc72RMe3SOMe, pyrSc7O45RMe, pyrSc7O45SMe, pyrSc73Me2, pyrSc7, pyrSaA3 butene, pyrSadA3 butene, dap7Gly, dap7PentDapAc7PDA, dap7Abu,4 vinylPyrSadV 3 butene, pyrSar 3 butene, pyrSap 3 butene, pyrSa4 Vie 2 Phe, or 3 allyl ySa.
In some embodiments, X 11 Is pyrS2. In some embodiments, X 11 Is Lys. In some embodiments, X 11 Is 3Thi. In some embodiments, X 11 Is Ala. In some embodiments, X 11 Is Phe. In some embodiments, X 11 Is S3MePyrSc7. In some embodiments, X 11 Is R3MePyrSc7. In some embodiments, X 11 Is S3iPrPyrSc7. In some embodiments, X 11 Is R3iPrPyrSc7.
In some embodiments, X 11 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-I, tables A-II, tables A-III and tables A-IV.
Various types of amino acid residues can be used for X 12 For example, amino acids of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or residues of salts thereof, according to the present disclosure. In some embodiments, X 12 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 12 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 12 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H.
In some embodiments, X 12 Comprising a side chain comprising an optionally substituted aromatic group. In some embodiments, X 12 Are aromatic amino acid residues as described herein. In some embodiments, the aromatic group is an optionally substituted 5-membered heteroaryl group having 1 to 3 heteroatoms. In some embodiments, the aromatic group is an optionally substituted 5-membered heteroaryl group having 1 to 3 nitrogen atoms. In some embodiments, the aromatic group is an optionally substituted 5-membered heteroaryl group having one oxygen atom. In some embodiments, the aromatic group is an optionally substituted 5-membered heteroaryl group having one sulfur atom. In some embodiments, the aromatic group is an optionally substituted 6 membered heteroaryl group having 1 to 3 heteroatoms. In some embodiments, the aromatic group is an optionally substituted 6 membered heteroaryl group having 1 nitrogen atom. In some embodiments, the aromatic group is an optionally substituted phenyl group. In some embodiments, X 12 Comprising a side chain which is OR comprises an optionally substituted aromatic group, wherein each substituent of the aromatic group is independently selected from halogen, -OR, -R, -C (O) OH, -C (O) NH 2 -CN or-NO 2 Wherein each R is independently C 1-4 Alkyl or haloalkyl. In some embodiments, the aromatic group is phenyl. In some embodiments, the aromatic group is an optionally substituted 8-to 10-membered bicyclic aryl or heteroaryl having 1 to 5 heteroatoms. In some embodiments, X 12 Comprising a side chain which is or comprises an optionally substituted aromatic group, wherein each substituent of the aromatic group is independently halogen. In some embodiments, X 12 Comprising a side chain which is or comprises two optionally substituted aromatic groups. In some embodiments, X 12 Comprising a side chain which is or comprises an optionally substituted aromatic group, wherein each of the aromatic groupsThe substituents are independently selected from halogen or-OH. In some embodiments, the aromatic group is phenyl. In some embodiments, the aromatic group is an optionally substituted 8-to 10-membered bicyclic aryl or heteroaryl having 0 to 5 heteroatoms. In some embodiments, the aromatic group is an optionally substituted 9-to 10-membered bicyclic aryl or heteroaryl having one heteroatom. In some embodiments, X 12 Are residues of amino acids of the formulae A-I or salts thereof. In some embodiments, the amino acid residue has a formula of-NH-C (R a2 )(R a3 ) -a C (O) -structure or a salt thereof. In some embodiments, the amino acid residue has the formula-NH-CH (R a3 ) -C) O) -structure or salt thereof. R, as described herein a3 is-L a -R', wherein each variable is independently as described herein. In some embodiments, R' is R, as described herein. In some embodiments, R is an optionally substituted group selected from the group consisting of: phenyl, 10 membered bicyclic aryl, 5 to 6 membered heteroaryl having 1 to 4 heteroatoms and 9 to 10 membered bicyclic heteroaryl having 1 to 5 heteroatoms. In some embodiments, each substituent is independently halogen or-OH or C 1-6 Halogenated aliphatic. In some embodiments, each substituent is independently halogen or-OH. In some embodiments, R is optionally substituted phenyl. In some embodiments, R is phenyl. In some embodiments, R is optionally substituted aryl. In some embodiments, R is aryl. In some embodiments, R is an optionally substituted 5 membered heteroaryl having 1 to 4 heteroatoms. In some embodiments, R is an optionally substituted 5 membered heteroaryl having 1 heteroatom. In some embodiments, optionally substituted R is a 6 membered heteroaryl having 1 to 4 heteroatoms. In some embodiments, optionally substituted R is a 6 membered heteroaryl having 1 heteroatom. In some embodiments, R is an optionally substituted 9-membered heteroaryl having 1 to 5 heteroatoms. In some embodiments, R is an optionally substituted 9-membered heteroaryl having 1 heteroatom. In some embodiments, R is an optionally substituted 10 membered heteroaryl having 1 to 5 heteroatoms. In some embodiments, R is optionally substituted with 1 hetero An atomic 10 membered heteroaryl. In some embodiments, the heteroatom is nitrogen. In some embodiments, the heteroatom is oxygen. In some embodiments, the heteroatom is sulfur. L as described herein a Is L. In some embodiments, L is a covalent bond. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution.
In some embodiments, X 12 Is a residue of an amino acid selected from the group consisting of: 3Thi,2F3MeF, phe,2COOHF,2ClF,2FurA,2OMeF,2MeF,2BrF,2CNF,2NO2F,2PyrA,3PyrA,4PyrA, his,1NapA,2Thi, and 2cmbF. In some embodiments, X 12 Is a residue of 3Thi,2F3MeF or Phe. In some embodiments, X 12 Is a residue of 3 Thi. In some embodiments, X 12 Is a residue of 2F3 MeF. In some embodiments, X 12 Is a residue of Phe. In some embodiments, X 12 Is a residue of 2 COOHF. In some embodiments, X 12 Is a residue of 2 ClF. In some embodimentsWherein X is 12 Is a residue of 2 FurA. In some embodiments, X 12 Is a residue of 2 OMeF. In some embodiments, X 12 Is a residue of 2 MeF. In some embodiments, X 12 Is a residue of 2 BrF. In some embodiments, X 12 Is a residue of 2 CNF. In some embodiments, X 12 Is a residue of 2NO 2F. In some embodiments, X 12 Is a residue of 2 pyraA. In some embodiments, X 12 Is a residue of 3 PyrA. In some embodiments, X 12 Is a residue of 4 pyrA. In some embodiments, X 12 Is a residue of His. In some embodiments, X 12 Is a residue of 1 NapA. In some embodiments, X 12 Is a residue of 2 Thi. In some embodiments, X 12 Is a residue of 2 cmbF. In some embodiments, 3Thi provides better properties and/or activity than, for example, phe.
In some embodiments, X 12 Is a residue of an amino acid whose side chain is hydrophobic. A variety of hydrophobic amino acid residues described herein can be used for X 12 For example for X 3 、X 7 Etc. as described. In some embodiments, X 12 Is a residue of nLeu, cypA, ala, leu, hLeu, npg, cpa, nva, cba, chA, val, ile, chg, hnLeu, or OctG. In some embodiments, X 12 Is a residue of nLeu or CypA. In some embodiments, X 12 Is a residue of nLeu. In some embodiments, X 12 Is a residue of CypA. In some embodiments, X 12 Is a residue of Ala. In some embodiments, X 12 Is a residue of Leu. In some embodiments, X 12 Is a residue of hLeu. In some embodiments, X 12 Is a residue of Npg. In some embodiments, X 12 Is a residue of Cpa. In some embodiments, X 12 Is a residue of Nva. In some embodiments, X 12 Is a residue of Cba. In some embodiments, X 12 Is a residue of ChA. In some embodiments, X 12 Is a residue of Val. In some embodiments, X 12 Is a residue of Ile. In some embodiments, X 12 Is a residue of Chg. In some embodiments, X 12 Is a residue of hnLeu. In some embodiments, X 12 Is a residue of OctG.
In some embodiments, X 12 Is the residue of an amino acid comprising an acidic or polar group. In some embodiments, X 12 Is a residue of an amino acid whose side chain contains an acidic group (e.g., -COOH group) or a salt form thereof (e.g., a compound of formula a-IV, etc.). A variety of acidic amino acid residues described herein can be used for X 12 For example for X 2 、X 5 、X 6 Etc. as described. In some embodiments, X 12 Is 2COOHF. In some embodiments, X 12 Is the residue of an amino acid whose side chain contains a polar group. In some embodiments, X 12 Is a compound whose side chain contains an amide group (e.g., -C (O) N (R') 2 For example-CONH 2 ) Residues of amino acids of (a). For example, in some embodiments, X 12 Is a residue of 2 cbmfs. A variety of other polar amino acid residues described herein can also be used for X 12 。
In some embodiments, X 12 Is a residue of an amino acid selected from the group consisting of: 3Thi,2F3MeF, phe, nLeu,2COOHF, cypA,2ClF, ala, abu, leu, hLeu, npg, cpa, nva, cba, chA,2FurA,2OMeF,2MeF,2BrF,2CNF,2NO2F,2PyrA,3PyrA,4PyrA, his,1NapA, val, ile, chg, diethA, hnLeu, octG,2Thi, and 2cmbF.
In some embodiments, X 12 Is that
3Thi, phe,2F3MeF, pyrS2,2ClF, hnLeu, bztA,2Thi,2MeF,2FF,34ClF, lys, nLeu,2COOHF,2PhF, hCbA, hCypA, hCHA, cypA, hpe, dipA, hepG, dap7Abu, hhLeu, hhSer, hexG, [2IAPAc ]2NH2F, ala, abu, leu, hleu, npg, cpa, pyrS1, [ Bnc ]2NH2F, [ Phc ]2NH2F, [ Biph ]2NH2F, [3PyAc ]2NH2F, nva, cba, chA,2 OMA, 2F, 2CNF,2 NOF, 2 yrA,3PyrA,4, pyrA, prA, pyrA, 2F, pmA, pyhPa, prA, P2F, pmGac, or [ PmC ]2NH2F, pcOc, pcF.
In some embodiments, X 12 Is 3Thi. In some embodiments, X 12 Is Phe. In some embodiments, X 12 Is 3F3MeF. In some embodiments, X 12 Is pyrS2. In some embodiments, X 12 Is 2ClF. In some embodiments, X 12 Is hnLeu. In some embodiments, X 12 Is BztA. In some embodiments, X 12 Is 2Thi. In some embodiments, X 12 Is 2MeF. In some embodiments, X 12 Is 2FF. In some embodiments, X 12 34ClF. In some embodiments, X 12 Is 2NH2F. In some embodiments, X 12 Is Trp. In some embodiments, X 12 Is 5ClW. In some embodiments, X 12 Is 6ClW. In some embodiments, X 12 Is 2NH2F. In some embodiments, X 12 Is [124TriAc ]]2NH2F. In some embodiments, X 12 Is [124TriPr]2NH2F. In some embodiments, X 12 Is [6QuiAc ]]2NH2F. In some embodiments, X 12 Is [2PyAc ]]2NH2F. In some embodiments, X 12 Is [2PyPrpc ]]2NH2F. In some embodiments, X 12 Is [3PyPrpc ]]2NH2F. In some embodiments, X 12 Is [4PyPrpc ]]2NH2F. In some embodiments, X 12 Is [ MeOPr ]]2NH2F. In some embodiments, X 12 Is [ PhOPr ]]2NH2F. In some embodiments, X 12 Is [ Me2MeOPr ]]2NH2F. In some embodiments, X 12 Is [ Me2NAc ]]2NH2F. In some embodiments, X 12 Is [ Me2NPr ]]2NH2F. In some embodiments, X 12 Is [ NdeMeButC]2NH2F. In some embodiments, X 12 Is [3IAPAc ] ]2NH2F. In some embodiments, X 12 Is [15PyraPy ]]2NH2F. In some embodiments, X 12 Is [ Morphac ]]2NH2F. In some embodiments, X 12 Is [ Nic ]]2NH2F. In some embodiments, X 12 Is [2PyzCO ]]2NH2F. In some embodiments, X 12 Is [5 pyrCO ]]2NH2F. In some embodiments, X 12 Is [3FPyr2c ]]2NH2F. In some embodiments, X 12 Is [4 ]FPyr3c]2NH2F。
In some embodiments, X 12 Are amino acid residues suitable for stapling as described herein. In some embodiments, X 12 For example with X 5 Stapling. In some embodiments, X 12 Is pyrS1. In some embodiments, X 12 Is pyrS2.
In some embodiments, X 12 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-IV.
In some embodiments, X 12 Interact with Trp383 of beta-catenin or its corresponding amino acid residue. In some embodiments, X 12 Interact with Asn415 of β -catenin or its corresponding amino acid residue. In some embodiments, X 12 Interact with Trp383 and Asn415 of β -catenin or their corresponding amino acid residues.
Various types of amino acid residues can be used for X 13 For example, amino acids of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or residues of salts thereof, according to the present disclosure. In some embodiments, X 13 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 13 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 13 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H.
In some embodiments, X 13 Comprising a side chain which is or comprises an optionally substituted aromatic group. In some embodiments, X 13 Are aromatic amino acid residues as described herein.
In some embodiments, X 13 Comprising a side containing an optionally substituted aromatic groupA chain. In some embodiments, X 13 Are aromatic amino acid residues as described herein. In some embodiments, the aromatic group is an optionally substituted 5-membered heteroaryl group having 1 to 3 heteroatoms. In some embodiments, the aromatic group is an optionally substituted 5-membered heteroaryl group having 1 to 3 nitrogen atoms. In some embodiments, the aromatic group is an optionally substituted 5-membered heteroaryl group having one sulfur atom. In some embodiments, the aromatic group is an optionally substituted phenyl group. In some embodiments, X 13 Comprising a side chain which is OR comprises an optionally substituted aromatic group, wherein each substituent of the aromatic group is independently selected from halogen, -OR, -R, -C (O) OH OR-CN, wherein each R is independently hydrogen OR C 1-4 Alkyl or haloalkyl. In some embodiments, the aromatic group is phenyl. In some embodiments, the aromatic group is an optionally substituted 8-to 10-membered bicyclic aryl or heteroaryl having 1 to 5 heteroatoms. In some embodiments, X 13 Comprising a side chain which is or comprises an optionally substituted aromatic group, wherein each substituent of the aromatic group is independently halogen. In some embodiments, X 13 Comprising a side chain which is or comprises two optionally substituted aromatic groups. In some embodiments, X 13 Comprising a side chain which is or comprises an optionally substituted aromatic group, wherein each substituent of the aromatic group is independently selected from halogen or-OH. In some embodiments, the aromatic group is phenyl. In some embodiments, the aromatic group is an optionally substituted 8-to 10-membered bicyclic aryl or heteroaryl having 0 to 5 heteroatoms. In some embodiments, the aromatic group is an optionally substituted 9-to 10-membered bicyclic aryl or heteroaryl having one heteroatom. In some embodiments, X 13 Is a residue of an amino acid of formula A-I or a salt thereof. In some embodiments, the amino acid residue has a formula of-NH-C (R a2 )(R a3 ) -a C (O) -structure or a salt thereof. In some embodiments, the amino acid residue has the formula-NH-CH (R a3 ) -C) O) -structure or salt thereof. R, as described herein a3 is-L a -R', wherein each variable is independently as described herein. In some embodiments, R' is R, as described herein. In some embodiments, R is an optionally substituted group selected from phenyl, 10 membered bicyclic aryl, 5 to 6 membered heteroaryl having 1 to 4 heteroatoms, and 9 to 10 membered bicyclic heteroaryl having 1 to 5 heteroatoms. In some embodiments, each substituent is independently halogen or-OH or C 1-6 Halogenated aliphatic. In some embodiments, each substituent is independently halogen or-OH. In some embodiments, R is optionally substituted phenyl. In some embodiments, R is phenyl. In some embodiments, R is optionally substituted aryl. In some embodiments, R is aryl. In some embodiments, R is an optionally substituted 5 membered heteroaryl having 1 to 4 heteroatoms. In some embodiments, R is an optionally substituted 5 membered heteroaryl having 1 heteroatom. In some embodiments, optionally substituted R is a 6 membered heteroaryl having 1 to 4 heteroatoms. In some embodiments, optionally substituted R is a 6 membered heteroaryl having 1 heteroatom. In some embodiments, R is an optionally substituted 9-membered heteroaryl having 1 to 5 heteroatoms. In some embodiments, R is an optionally substituted 9-membered heteroaryl having 1 heteroatom. In some embodiments, R is an optionally substituted 10 membered heteroaryl having 1 to 5 heteroatoms. In some embodiments, R is an optionally substituted 10 membered heteroaryl having 1 heteroatom. In some embodiments, the heteroatom is nitrogen. In some embodiments, the heteroatom is oxygen. In some embodiments, the heteroatom is sulfur. L as described herein a Is L. In some embodiments, L is a covalent bond. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 A hydrocarbon chain. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is a divalent straight or branched chain C 1-10 A hydrocarbon chain. In some implementationsIn embodiments, L is a divalent straight chain C 1-10 A hydrocarbon chain. In some embodiments, L is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is- (CH) 2 ) n-, wherein n is 1 to 10. In some embodiments, L is-CH 2 -. In some embodiments, L is- (CH) 2 ) 2 -. In some embodiments, L is- (CH) 2 ) 3 -. In some embodiments, L is- (CH) 2 ) 4 -. In some embodiments, L is an optionally substituted divalent straight or branched chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution. In some embodiments, L is an optionally substituted divalent straight chain C 1-10 Hydrocarbon chains in which one or more methylene units of L are independently replaced by-C (R') 2 -C (O) -, -N (R'), -Cy-, or-O-substitution.
In some embodiments, X 13 Is a residue of BztA, 34ClF or 2 NapA. In some embodiments, X 13 Is a residue of BztA. In some embodiments, X 13 Is a residue of 34ClF. In some embodiments, X 13 Is a residue of 2NapA. In some embodiments, X 13 Is a residue of 3 BrF. In some embodiments, X 13 Is a residue of 3Thi. In some embodiments, X 13 Is a residue of 34MeF.
In some embodiments, X 13 Is BztA,34ClF,3Thi,Phe,GlnR,34MeF,2NapA,Lys,PyrS2,3BrF,7FBztA,2BrF,3F3MeF,4F3MeF,RbMe2NapA,RbMeBzta,SbMeBzta,5IndA,7ClBztA,7MeBztA,Leu,2ClF,3ClF,4BrF,4ClF, or 3MeF.
In some embodiments, X 13 Is BztA. In some embodiments, X 13 34ClF. In some embodiments, X 13 Is 3Thi. In some embodiments, X 13 Is Phe. In some embodiments, X 13 Is GlnR. In some embodiments, X 13 34MeF. In some embodiments, X 13 Is 2NapA. In some casesIn embodiments, X 13 Is Lys. In some embodiments, bztA provides better properties and/or activity than, for example, trp.
In some embodiments, X 13 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-IV.
In some embodiments, X 13 Interact with Gln379 of β -catenin or its corresponding amino acid residue. In some embodiments, X 13 Interact with Leu382 of beta-catenin or its corresponding amino acid residue. In some embodiments, X 13 Interact with Val416 of β -catenin or its corresponding amino acid residue. In some embodiments, X 13 Interact with Asn415 of β -catenin or its corresponding amino acid residue. In some embodiments, X 13 Interact with Trp383 of beta-catenin or its corresponding amino acid residue. In some embodiments, X 13 Interact with Gln379, leu382, val416, asn415 and Trp383 of beta-catenin or their corresponding amino acid residues.
Various types of amino acid residues can be used for X 14 For example, amino acids of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or residues of salts thereof, according to the present disclosure. In some embodiments, X 14 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 14 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 14 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H.
In some embodiments, X 14 Are amino acid residues suitable for stapling. In some embodiments, X 14 Is stapled. In some embodiments, X 14 And X is 10 Stapling, as herein describedSaid. In some embodiments, X 14 And X is 7 Stapling, as described herein.
In some embodiments, X 14 Amino acid residues suitable for stapling, e.g. for X 7 、X 10 Etc. as described.
Various types of amino acid residues can be used for X 14 . In some embodiments, X 14 Is GlnR, lys, sA, gln, cys, triAzLys, asnR, hGlnR,4PipA,sAbu,Orn,dGlnR, [4 mammoperidine ]]GlnR, [39N2 spiroundecane]GlnR, [29N2 spiroundecane]GlnR, iPrLys, sCH2S, [ diaminobutane ]]GlnR, or [4 aminopiperidine]GlnR。
In some embodiments, X 14 Is GlnR. In some embodiments, X 14 Is Lys. In some embodiments, X 14 Is sA. In some embodiments, X 14 Is Gln. In some embodiments, X 14 Is Cys. In some embodiments, X 14 Is TriAzLys. In some embodiments, X 14 Is AsnR. In some embodiments, X 14 Is hGLnR. In some embodiments, X 14 Is 4PipA. In some embodiments, X 14 Is sAbu. In some embodiments, X 14 Is Orn. In some embodiments, X 14 Is dGlnR. In some embodiments, X 14 Is [4 mamplatidine ]]GlnR. In some embodiments, X 14 Is [39N2 spiroundecane]GlnR. In some embodiments, X 14 Is [29N2 spiroundecane]GlnR. In some embodiments, X 14 Is iPrLys. In some embodiments, X 14 sCH2S. In some embodiments, X 14 Is [ diaminobutane ]]GlnR. In some embodiments, X 14 Is [4 aminopiperidine]GlnR。
In some embodiments, X 14 Are aromatic amino acid residues as described herein. In some embodiments, X 14 Is BtzA.
In some embodiments, v14 is a polar amino acid residue as described herein. In some implementationsIn embodiments, X 14 Is Gln.
In some embodiments, X 14 Is a C-terminal amino acid residue. In some embodiments, X 14 Having free-COOH or a salt form thereof. In some embodiments, X 14 and-C (O) OH is capped. In some embodiments, X 14 Conversion of-C (O) OH to-C (O) N (R') 2 Wherein each R is independently as described herein. In some embodiments, -C (O) N (R') 2 is-C (O) NHR'. In some embodiments, each R' is independently R. In some embodiments, each R' is-H. In some embodiments, R is H. In some embodiments, R is optionally substituted C 1-6 Aliphatic series. In some embodiments, R is optionally substituted C 1-6 An alkyl group. In some embodiments, R is ethyl. In some embodiments, R isIn some embodiments, R is-CH (CH 3 )CH 2 OH. In some embodiments, R is- (S) -CH (CH) 3 )CH 2 OH. In some embodiments, R is- (R) -CH (CH) 3 )CH 2 OH. In some embodiments, R is-CH (CH 2 OH) 2 。
In some embodiments, two R' groups together with the nitrogen atom to which they are attached form a ring, as described herein. In some embodiments, -N (R') 2 Is that
In some embodiments, X 14 Is that
GlnR, bztA, sAla,34ClF, cys, ala, lys, asnR, aMeC, pyrS2, gln, hGLN R,3Thi, lys, pen, glnR, triAzLys, hCys,4PipA, sAbu, orn,1MeK, [4 maleeridine ] GlnR, [39N2 spiroundecane ] GlnR, [29N2 spiroundecane ] GlnR, iPrLys, sCH S, asnEDA, asnS3APyr, [ diaminobutane ] GlnR, [4 aminopiperidine ] GlnR, dGlnR, glnEDA, asnPpz, glnPpz, glnR3APyr, glnS3APyr, glnME2EDA, asnME2EDA, asnMeEDA, asnR APyr.
In some embodiments, X 14 Is GlnR. In some embodiments, X 14 Is BztA. In some embodiments, X 14 Is sA. In some embodiments, X 14 34ClF. In some embodiments, X 14 Is Cys. In some embodiments, X 14 Is Ala. In some embodiments, X 14 Is Lys. In some embodiments, X 14 Is AsnR. In some embodiments, X 14 Is aMeC. In some embodiments, X 14 Is pyrS2. In some embodiments, X 14 Containing a C-terminal group, such as-NH 2. In some embodiments, X 14 Is Gln. In some embodiments, X 14 Is hGLnR. In some embodiments, X 14 Is 3Thi. In some embodiments, X 14 Is Lys. In some embodiments, X 14 Is GlnR 3. In some embodiments, X 14 Is dLys. In some embodiments, X 14 Is GlnMEPDA. In some embodiments, X 14 Is GlnT4CyMe. In some embodiments, X 14 Is GlnMEBDA. In some embodiments, X 14 Is Gln5DA. In some embodiments, X 14 Is Gln6DA. In some embodiments, X 14 Is TriAzOrn. In some embodiments, X 14 Is Phe. In some embodiments, X 14 Is GlnC4CyMe. In some embodiments, X 14 Is Gln3ACPIP. In some embodiments, X 14 Is GlnPipAz. In some embodiments, X 14 Is GlnPip4AE. In some embodiments, X 14 Forming intramolecular hydrogen bonds.
In some embodiments, X 14 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-I, tables A-II, tables A-III and tables A-IV.
In some embodiments, p15 is 1. In some embodiments, p15 is 0.
Various types of amino acid residues can be used for X 15 For example, the first and second substrates may be coated, for example,residues of amino acids of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or salts thereof, according to the present disclosure. In some embodiments, X 15 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 15 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 15 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H.
Various types of amino acid residues can be used for X 15 . In some embodiments, X 15 Ala, leu, val, aib, morphNva, thr, dAla, dLeu, [ biotin PEG8 ] ]Residues of Lys, glu or AzLys.
In some embodiments, X 15 Is or comprises a label, e.g., a label for detection, binding, etc. In some embodiments, the label is or comprises biotin. In some embodiments, X 15 Is [ biotin PEG8 ]]Lys。
In some embodiments, X 15 Are hydrophobic amino acid residues as described herein, e.g., for X 3 、X 8 Etc. as described herein. In some embodiments, X 15 Is Ala. In some embodiments, X 15 Is Leu. In some embodiments, X 15 Is Val. In some embodiments, X 15 Is Aib. In some embodiments, X 15 Is dAba. In some embodiments, X 15 Is dLeu.
In some embodiments, X 15 Is an amino acid residue whose side chain contains an amino group. In some embodiments, X 15 Is MorphNva.
In some embodiments, X 15 Are amino acid residues for stapling, as described herein. In some embodiments, X 15 Is GlnR. In some embodiments, it is in combination with X 11 Stapling. In some embodiments, X 11 Is Lys.
In some embodiments, X 15 Are polar amino acid residues as described herein, e.g., for X 2 、X 5 、X 6 Etc. as described herein. In some embodiments, X 15 Is Thr. In some embodiments, X 15 is-Ser.
In some embodiments, X 15 Are acidic amino acid residues as described herein, e.g., for X 2 、X 5 、X 6 Etc. as described herein. In some embodiments, X 15 Is Glu.
In some embodiments, X 15 Is a C-terminal amino acid residue. In some embodiments, X 15 Having free-COOH or a salt form thereof. In some embodiments, X 15 and-C (O) OH is capped. In some embodiments, X 15 Conversion of-C (O) OH to-C (O) N (R') 2 Wherein each R is independently as described herein. In some embodiments, -C (O) N (R') 2 is-C (O) NHR'. In some embodiments, each R' is independently R. In some embodiments, each R' is-H. In some embodiments, R is H. In some embodiments, R is optionally substituted C 1-6 Aliphatic series. In some embodiments, R is optionally substituted C 1-6 An alkyl group. In some embodiments, R is ethyl. In some embodiments, R isIn some embodiments, R is-CH (CH 3 )CH 2 OH. In some embodiments, R is- (S) -CH (CH) 3 )CH 2 OH. In some embodiments, R is- (R) -CH (CH) 3 )CH 2 OH. In some embodiments, R is-CH (CH 2 OH) 2 。
In some embodiments, the agent comprises a C-terminal group. In some embodiments, the C-terminal group is-OH. In some embodiments, the C terminal groupThe radical being-NH 2 。
In some embodiments, X 15 Is Ala, glnR, leu, val, ser, thr,3Thi, bztA, aib, morphNva, dA, dLeu, pro, phe, [ biotin PEG8 ]]Lys，Throl，Glu，AzLys，Npg，Trp，Tyr，Lys，Prool，Alaol，Gly，dPro，Asn，Gln，Ala_D3，[mPEG4]Lys，[mPEG8]Lys，[mPEG16]Lys。
In some embodiments, X 15 Is Ala. In some embodiments, X 15 Containing C-terminal groups, e.g. -NH 2 . In some embodiments, X 15 Is GlnR. In some embodiments, X 15 Is Leu. In some embodiments, X 15 Is Val. In some embodiments, X 15 Is Ser. In some embodiments, X 15 Is Thr. In some embodiments, X 15 Is 3Thi. In some embodiments, X 15 Is BztA. In some embodiments, X 15 Is [ mPEG37 ]]-Lys. In some embodiments, X 15 Is dVal. In some embodiments, X 15 34ClF.
In some embodiments, X 15 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-IV.
In some embodiments, p16 is 1. In some embodiments, p16 is 0.
Various types of amino acid residues can be used for X 16 For example, amino acids of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or residues of salts thereof, according to the present disclosure. In some embodiments, X 16 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 16 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 16 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodimentsWherein R is a3 is-H.
Various types of amino acid residues can be used for X 16 . In some embodiments, X 16 Is a residue of Ser, ala, glu, aib, asp, thr or aThr.
In some embodiments, X 16 Are polar amino acid residues as described herein, e.g., for X 2 、X 5 、X 6 Etc. as described herein. In some embodiments, X 16 Is Thr. In some embodiments, X 16 is-Ser. In some embodiments, X 16 Is aThr.
In some embodiments, X 16 Are hydrophobic amino acid residues as described herein, e.g., for X 3 、X 8 Etc. as described herein. In some embodiments, X 16 Is Ala. In some embodiments, X 16 Is Leu. In some embodiments, X 16 Is Val. In some embodiments, X 16 Is Aib. In some embodiments, X 16 Is dAba. In some embodiments, X 16 Is dLeu.
In some embodiments, X 16 Are acidic amino acid residues as described herein, e.g., for X 2 、X 5 、X 6 Etc. as described herein. In some embodiments, X 16 Is Glu. In some embodiments, X 16 Is Asp.
In some embodiments, X 16 Is Ala, ser, glu, glnR, bztA, thr, aib, asp, lys, aThr, val or Arg. In some embodiments, X 16 Containing C-terminal groups, e.g. NH 2 OH, serol, NHEt, NHMe, dAlaol, etc.
In some embodiments, X 16 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-IV.
In some embodiments, p17 is 1. In some embodiments, p17 is 0.
Various types of amino acid residues can be used for X 17 For example, according to the present disclosure, formula A-I, A-II,Residues of amino acids A-III, A-IV, A-V, A-VI, etc., or salts thereof. In some embodiments, X 17 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 17 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 17 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H.
In some embodiments, X 17 Are hydrophobic amino acid residues as described herein, e.g., for X 3 、X 8 Etc. as described herein. In some embodiments, X 17 Is a residue of Ala or Leu. In some embodiments, X 17 Is a residue of Ala. In some embodiments, X 17 Is a residue of Leu.
In some embodiments, X 17 Ala, leu, glnR, glnR, pro, thr, val, lys, arg, [ Ac ]]Lys、[mPEG4]Lys、[mPEG8]Lys or [ mPEG16 ]]Lys. In some embodiments, X 17 Containing C-terminal groups, e.g. NH 2 NHEt, OH, etc. In some embodiments, X 17 Is [ Ac-dPEG2 ]]-Lys. In some embodiments, X 17 Is [ Ac-PEG8 ]]-Lys. In some embodiments, X 17 Is [ Oct-dPEG2]-Lys. In some embodiments, X 17 Is [ Oct-PEG8 ]]-Lys. In some embodiments, X 17 Is [ C18-dPEG2 ]]-Lys. In some embodiments, X 17 Is [ C18-PEG8 ]]-Lys. In some embodiments, X 17 Is [ Adamant C-dPEG2 ]]-Lys. In some embodiments, X 17 Is [ Adamant C-PEG8 ]]-Lys. In some embodiments, X 17 Is [ lithocholic acid-dPEG 2 ]]-Lys. In some embodiments, X 17 Is [ lithocholic acid-PEG 8 ]]-Lys。
In some embodiments, X 17 Is or comprises the following: selected from tables A-IVResidues of partial or amino acids.
In some embodiments, X 17 Comprising polar side chains. In some embodiments, it is a polar amino acid residue as described herein. In some embodiments, X 17 Comprising nonpolar side chains. In some embodiments, X 17 Comprising hydrophobic side chains. In some embodiments, it is a hydrophobic amino acid residue as described herein. In some embodiments, X 17 Comprising aliphatic side chains. In some embodiments, X 17 Comprising alkyl side chains. In some embodiments, X 17 Is C as a side chain 1-10 An alkyl group. In some embodiments, X 17 Comprising a side chain containing an optionally substituted aromatic group. In some embodiments, it is an aromatic amino acid residue as described herein. In some embodiments, X 17 Comprising a side chain containing an acidic group (e.g., -COOH). In some embodiments, it is an acidic amino acid residue as described herein. In some embodiments, X 17 Comprising groups containing basic groups (e.g., -N (R) 2 ) Is a side chain of (c). In some embodiments, it is a basic amino acid residue as described herein. In some embodiments, X 17 Comprising a detectable moiety, such as a fluorescent moiety. In some embodiments, X 17 Is Ala, dAla or Leu. In some embodiments, X 17 Is Ala. In some embodiments, X 17 Is dAba. In some embodiments, X 17 Is Leu.
In some embodiments, X 17 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-IV.
In some embodiments, p17 is 1. In some embodiments, p17 is 0.
Various types of amino acid residues can be used for X 18 For example, amino acids of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or residues of salts thereof, according to the present disclosure. In some embodiments, X 18 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)-wherein each variable is independently as described herein. In some embodiments, X 18 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 18 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H.
In some embodiments, X 18 Comprising polar side chains. In some embodiments, it is a polar amino acid residue as described herein. In some embodiments, X 18 Comprising nonpolar side chains. In some embodiments, X 18 Comprising hydrophobic side chains. In some embodiments, it is a hydrophobic amino acid residue as described herein. In some embodiments, X 18 Comprising aliphatic side chains. In some embodiments, X 18 Comprising alkyl side chains. In some embodiments, X 18 Is C as a side chain 1-10 An alkyl group. In some embodiments, X 18 Comprising a side chain containing an optionally substituted aromatic group. In some embodiments, it is an aromatic amino acid residue as described herein. In some embodiments, X 18 Comprising a side chain containing an acidic group (e.g., -COOH). In some embodiments, it is an acidic amino acid residue as described herein. In some embodiments, X 18 Comprising groups containing basic groups (e.g., -N (R) 2 ) Is a side chain of (c). In some embodiments, it is a basic amino acid residue as described herein. In some embodiments, X 18 Comprising a detectable moiety, such as a fluorescent moiety. In some embodiments, X 18 Is Aib, ala or Leu. In some embodiments, X 18 Is Ala or Leu. In some embodiments, X 18 Is Aib. In some embodiments, X 18 Is Ala. In some embodiments, X 18 Is Leu. In some embodiments, X 18 Is Pro. In some embodiments, X 18 Is [ Ac ]]Lys. In some embodiments，X 18 Is [ mPEG4 ]]Lys. In some embodiments, X 18 Is [ mPEG8 ]]Lys. In some embodiments, X 18 Is [ mPEG16 ]]Lys. In some embodiments, X 18 Is Thr. In some embodiments, X 18 Is GlnR. In some embodiments, X 18 Is [ mPEG37 ]]Lys. In some embodiments, X 18 Is [ PEG4triPEG16 ]]Lys. In some embodiments, X 18 Is [ PEG4triPEG36 ]]Lys. In some embodiments, X 18 Comprising a C-terminal group as described herein.
In some embodiments, X 18 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-IV.
In some embodiments, p18 is 1. In some embodiments, p18 is 0.
Various types of amino acid residues can be used for X 19 For example, amino acids of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or residues of salts thereof, according to the present disclosure. In some embodiments, X 19 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 19 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 19 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H.
In some embodiments, X 19 Comprising polar side chains. In some embodiments, it is a polar amino acid residue as described herein. In some embodiments, X 19 Comprising nonpolar side chains. In some embodiments, X 19 Comprising hydrophobic side chains. In some embodiments, it is a hydrophobic amino acid residue as described herein. In some embodiments, X 19 Comprising aliphatic side chains. In some embodiments, X 19 IncludedAlkyl side chains. In some embodiments, X 19 Is C as a side chain 1-10 An alkyl group. In some embodiments, X 19 Comprising a side chain containing an optionally substituted aromatic group. In some embodiments, it is an aromatic amino acid residue as described herein. In some embodiments, X 19 Comprising a side chain containing an acidic group (e.g., -COOH). In some embodiments, it is an acidic amino acid residue as described herein. In some embodiments, X 19 Comprising groups containing basic groups (e.g., -N (R) 2 ) Is a side chain of (c). In some embodiments, it is a basic amino acid residue as described herein. In some embodiments, X 19 Comprising a detectable moiety, such as a fluorescent moiety. In some embodiments, X 19 Is Aib, ala or Leu. In some embodiments, X 19 Is Ala or Leu. In some embodiments, X 19 Is Aib. In some embodiments, X 19 Is Ala. In some embodiments, X 19 Is Leu. In some embodiments, X 19 Is Thr. In some embodiments, X 19 Is Val. In some embodiments, X 19 Is Pro.
In some embodiments, X 19 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-IV.
In some embodiments, p19 is 1. In some embodiments, p19 is 0.
Various types of amino acid residues can be used for X 20 For example, amino acids of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or residues of salts thereof, according to the present disclosure. In some embodiments, X 20 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 20 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 20 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently asAs described herein. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H.
In some embodiments, X 20 Comprising polar side chains. In some embodiments, it is a polar amino acid residue as described herein. In some embodiments, X 20 Comprising nonpolar side chains. In some embodiments, X 20 Comprising hydrophobic side chains. In some embodiments, it is a hydrophobic amino acid residue as described herein. In some embodiments, X 20 Comprising aliphatic side chains. In some embodiments, X 20 Comprising alkyl side chains. In some embodiments, X 20 Is C as a side chain 1-10 An alkyl group. In some embodiments, X 20 Comprising a side chain containing an optionally substituted aromatic group. In some embodiments, it is an aromatic amino acid residue as described herein. In some embodiments, X 20 Comprising a side chain containing an acidic group (e.g., -COOH). In some embodiments, it is an acidic amino acid residue as described herein. In some embodiments, X 20 Comprising groups containing basic groups (e.g., -N (R) 2 ) Is a side chain of (c). In some embodiments, it is a basic amino acid residue as described herein. In some embodiments, X 20 Comprising a detectable moiety, such as a fluorescent moiety. In some embodiments, X 20 Is Aib, ala or Leu. In some embodiments, X 20 Is Ala or Leu. In some embodiments, X 20 Is Aib. In some embodiments, X 20 Is Ala. In some embodiments, X 20 Is Leu. In some embodiments, X 20 Is Lys. In some embodiments, X 20 Is nLeu. In some embodiments, X 20 Is Val. In some embodiments, X 20 Is Arg.
In some embodiments, X 20 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-IV.
In some embodiments, p20 is 1. In some embodiments, p20 is 0.
Various types of amino acid residues can be used for X 21 For example, amino acids of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or residues of salts thereof, according to the present disclosure. In some embodiments, X 21 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 21 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 21 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H.
In some embodiments, X 21 Comprising polar side chains. In some embodiments, it is a polar amino acid residue as described herein. In some embodiments, X 21 Comprising nonpolar side chains. In some embodiments, X 21 Comprising hydrophobic side chains. In some embodiments, it is a hydrophobic amino acid residue as described herein. In some embodiments, X 21 Comprising aliphatic side chains. In some embodiments, X 21 Comprising alkyl side chains. In some embodiments, X 21 Is C as a side chain 1-10 An alkyl group. In some embodiments, X 21 Comprising a side chain containing an optionally substituted aromatic group. In some embodiments, it is an aromatic amino acid residue as described herein. In some embodiments, X 21 Comprising a side chain containing an acidic group (e.g., -COOH). In some embodiments, it is an acidic amino acid residue as described herein. In some embodiments, X 21 Comprising groups containing basic groups (e.g., -N (R) 2 ) Is a side chain of (c). In some embodiments, it is a basic amino acid residue as described herein. In some embodiments, X 21 Comprising a detectable moiety, such as a fluorescent moiety. In some embodiments, X 21 Is Aib, ala orLeu. In some embodiments, X 21 Is Ala or Leu. In some embodiments, X 21 Is Aib. In some embodiments, X 21 Is Ala. In some embodiments, X 21 Is Leu. In some embodiments, X 21 Is Lys. In some embodiments, X 21 Is nLeu. In some embodiments, X 21 Is Arg.
In some embodiments, X 21 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-IV.
In some embodiments, p21 is 1. In some embodiments, p21 is 0.
Various types of amino acid residues can be used for X 22 For example, amino acids of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or residues of salts thereof, according to the present disclosure. In some embodiments, X 22 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 22 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 22 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H.
In some embodiments, X 22 Comprising polar side chains. In some embodiments, it is a polar amino acid residue as described herein. In some embodiments, X 22 Comprising nonpolar side chains. In some embodiments, X 22 Comprising hydrophobic side chains. In some embodiments, it is a hydrophobic amino acid residue as described herein. In some embodiments, X 22 Comprising aliphatic side chains. In some embodiments, X 22 Comprising alkyl side chains. In some embodiments, X 22 Is C as a side chain 1-10 An alkyl group. In some embodiments, X 22 Comprises optionally containingSide chains of substituted aromatic groups. In some embodiments, it is an aromatic amino acid residue as described herein. In some embodiments, X 22 Comprising a side chain containing an acidic group (e.g., -COOH). In some embodiments, it is an acidic amino acid residue as described herein. In some embodiments, X 22 Comprising groups containing basic groups (e.g., -N (R) 2 ) Is a side chain of (c). In some embodiments, it is a basic amino acid residue as described herein. In some embodiments, X 22 Comprising a detectable moiety, such as a fluorescent moiety. In some embodiments, X 22 Is Aib, ala or Leu. In some embodiments, X 22 Is Ala or Leu. In some embodiments, X 22 Is Aib. In some embodiments, X 22 Is Ala. In some embodiments, X 22 Is Leu. In some embodiments, X 22 Is Lys.
In some embodiments, X 22 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-IV.
In some embodiments, p22 is 1. In some embodiments, p22 is 0.
Various types of amino acid residues can be used for X 23 For example, amino acids of formulas A-I, A-II, A-III, A-IV, A-V, A-VI, etc., or residues of salts thereof, according to the present disclosure. In some embodiments, X 23 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 23 is-N (R) a1 )-C(R a2 )(R a3 ) -C (O) -, wherein each variable is independently as described herein. In some embodiments, X 23 is-N (R) a1 )-C(R a2 ) H-C (O) -, wherein each variable is independently as described herein. In some embodiments, R a1 is-H. In some embodiments, R a3 is-H.
In some embodiments, X 23 Comprising polar side chains. In some embodiments, it is asPolar amino acid residues as described herein. In some embodiments, X 23 Comprising nonpolar side chains. In some embodiments, X 23 Comprising hydrophobic side chains. In some embodiments, it is a hydrophobic amino acid residue as described herein. In some embodiments, X 23 Comprising aliphatic side chains. In some embodiments, X 23 Comprising alkyl side chains. In some embodiments, X 23 Is C as a side chain 1-10 An alkyl group. In some embodiments, X 23 Comprising a side chain containing an optionally substituted aromatic group. In some embodiments, it is an aromatic amino acid residue as described herein. In some embodiments, X 23 Comprising a side chain containing an acidic group (e.g., -COOH). In some embodiments, it is an acidic amino acid residue as described herein. In some embodiments, X 23 Comprising groups containing basic groups (e.g., -N (R) 2 ) Is a side chain of (c). In some embodiments, it is a basic amino acid residue as described herein. In some embodiments, X 23 Comprising a detectable moiety, such as a fluorescent moiety. In some embodiments, X 23 Is Aib, ala or Leu. In some embodiments, X 23 Is Ala or Leu. In some embodiments, X 23 Is Aib. In some embodiments, X 23 Is Ala. In some embodiments, X 23 Is Leu.
In some embodiments, X 23 Is or comprises the following: residues selected from the group consisting of the moieties or amino acids of tables A-IV.
In some embodiments, p23 is 1. In some embodiments, p23 is 0.
In some embodiments, the agent is or comprises a peptide having the structure or a salt thereof:
R N -[X] p -[X 0 ] p0 X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 -[X] p’ -R C ，
wherein:
each X is independently an amino acid residue;
each p and p' is independently 0 to 10;
R N independently a peptide, amino protecting group or R' -L RN -；
R C Independently a peptide, a carboxyl protecting group, -L RC -R’、-O-L RC -R 'or-N (R') -L RC -R’；
L RN And L RC Each independently is L;
each L is independently a covalent bond or an optionally substituted divalent C 1 -C 25 Aliphatic or heteroaliphatic groups having from 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R')-, -C (O) S-or-C (O) O-;
each-Cy-is independently an optionally substituted divalent 3-to 30-membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms;
each R' is independently-R, -C (O) R, -CO 2 R or-SO 2 R；
Each R is independently-H or an optionally substituted group selected from: c (C) 1-30 Aliphatic, C having 1 to 10 heteroatoms 1-30 Heteroaliphatic, C 6-30 Aryl, C 6-30 Arylaliphatic, C having 1 to 10 heteroatoms 6-30 Aryl heteroaliphatics, 5 to 30 membered heteroaryl groups having 1 to 10 heteroatoms and 3 to 30 membered heterocyclyl groups having 1 to 10 heteroatoms, or
The two R groups optionally and independently together form a covalent bond, or:
two or more R groups on the same atom optionally and independently taken together with the atom form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the atom; or alternatively
Two or more R groups on two or more atoms optionally and independently together with their intervening atoms form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the intervening atoms.
In some embodiments, p is 0. In some embodiments, p is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, p is 1. In some embodiments, p is 2. In some embodiments, p is 3. In some embodiments, p is 4. In some embodiments, p is 5. In some embodiments, p is 6. In some embodiments, p is 7. In some embodiments, p is 8. In some embodiments, p is 9. In some embodiments, p is 10.
In some embodiments, p' is 0. In some embodiments, p' is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, p' is 1. In some embodiments, p' is 2. In some embodiments, p' is 3. In some embodiments, p' is 4. In some embodiments, p' is 5. In some embodiments, p' is 6. In some embodiments, p' is 7. In some embodiments, p' is 8. In some embodiments, p' is 9. In some embodiments, p' is 10.
In some embodiments, R N Is an N-terminal capping group. In some embodiments, R N is-C (O) R, wherein R is as described herein. In some embodiments, R is-H. In some embodiments, R is optionally substituted C 1-6 Aliphatic series. In some embodiments, R is optionally substituted C 1-6 An alkyl group. In some embodiments, R is methyl. In some embodiments, R N Is Ac. In some embodiments, R N Is a group suitable for stapling or stapling.In some embodiments, R N Is 4 pentenyl. In some embodiments, R N Is 5 hexenyl. In some embodiments, R N Is BzAm2OAllyl. In some embodiments, R N Is Ac, NPyroR3,5 hexenyl, 4 pentenyl, bua, C3a, cpc, cbc, cypCO, bnc, CF3CO,2PyCypCO,4THPCO, isobutyryl, TTs,15PyraPy,2PyBu,4PymCO,4PyPrpc,3IAPAc,4MePipzPrpC,MePipAc,MeImid4SO2,BzAm2OAllyl,Hex,2PyzCO,3Phc3,MeOPr, lithocholate, 2FPhc, phC, meSO2, isovaleryl, etHNCO, tzPyr,8IAP,3PydCO,2PymCO,5PymCO,1Imidac,2F2PyAc,2IAPAc,124TriPr,6QuiAc,3PyAc,123TriAc,1PyrazoleAc,3PyPrpc,5PymAc,1PydoneAc,124TriAc,Me2NAc,8QuiSO2,mPEG4,mPEG8,mPEG16 or mPEG24.
In some embodiments, R C Is a C-terminal capping group. In some embodiments, R C is-N (R') 2 Wherein each R' is independently as described herein. In some embodiments, R C is-NHR ', wherein R' is as described herein. In some embodiments, R C is-N (R) 2 Wherein each R is independently as described herein. In some embodiments, R C is-NHR, wherein R is as described herein. In some embodiments, R is-H. In some embodiments, R is optionally substituted C 1-6 Aliphatic series. In some embodiments, R is optionally substituted C 1-6 An alkyl group. In some embodiments, R is methyl. In some embodiments, R is ethyl. In some embodiments, R C is-NH 2 . In some embodiments, R C is-NHEt.
In some embodiments, R C is-NHC (CH) 3 )CH 2 OH. In some embodiments, R C Is- (S) -NHC (CH 3 )CH 2 OH. In some embodiments, R C Is- (R) -NHC (CH) 3 )CH 2 OH. In some embodiments, R C Is thatIn some embodiments, R C Is->In some embodiments, R C Is->In some embodiments, R C Is->In some embodiments, R C Is->
In some embodiments, R C is-Alaol, wherein the amino group of-Alaol is bonded with the last-C (O) -of the peptide skeleton (R C Is that). In some embodiments, R C is-dAlol, wherein the amino group of-dAlol is bonded with the last-C (O) -of the peptide backbone (R C Is->). In some embodiments, R C is-Prool, wherein the amino group of-Prool is bonded to the last-C (O) -of the peptide backbone (R C Is->). In some embodiments, R C is-Throol, wherein the amino group of-Throol is bonded to the last-C (O) -of the peptide backbone (R C Is->). In some embodiments, R C is-Serol, wherein the amino group of-Serol is bonded to the last-C (O) -of the peptide backbone (R C Is->)。
In some embodiments, R C is-OH.
Amino acids
As will be appreciated by those of skill in the art, a variety of amino acids may be used in accordance with the present disclosure. For example, both naturally occurring and non-naturally occurring amino acids may be used in accordance with the present disclosure. In some embodiments, the amino acid is a compound comprising an amino group that can form an amide group with a carboxyl group and a carboxyl group. In some embodiments, the amino acid is an alpha-amino acid. In some embodiments, the amino acid is a β -amino acid. In some embodiments, the amino acid is a D-amino acid. In some embodiments, the amino acid is an L-amino acid. In some embodiments, the amino acid is a naturally encoded amino acid, e.g., an amino acid in a mammalian cell.
In some embodiments, the amino acid is a compound having the structure of formula a-I:
N(R a1 ) 2 -L a1 -C(R a2 )(R a3 )-L a2 -COOH,
A-I
wherein:
R a1 、R a2 、R a3 each independently is-L a -R’；
L a 、L a1 And L a2 Each independently is L;
each L is independently a covalent bond or an optionally substituted divalent C 1 -C 25 Aliphatic or heteroaliphatic groups having from 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R')-, -C (O) S-or-C (O) O-;
each-Cy-is independently an optionally substituted divalent 3-to 30-membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms;
each R' is independently-R, -C (O) R, -CO 2 R or-SO 2 R；
Each R is independently-H or an optionally substituted group selected from: c (C) 1-30 Aliphatic, C having 1 to 10 heteroatoms 1-30 Heteroaliphatic, C 6-30 Aryl, C 6-30 Arylaliphatic, C having 1 to 10 heteroatoms 6-30 Aryl heteroaliphatics, 5 to 30 membered heteroaryl groups having 1 to 10 heteroatoms and 3 to 30 membered heterocyclyl groups having 1 to 10 heteroatoms, or
The two R groups optionally and independently together form a covalent bond, or:
two or more R groups on the same atom optionally and independently taken together with the atom form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the atom; or alternatively
Two or more R groups on two or more atoms optionally and independently together with their intervening atoms form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the intervening atoms.
In some embodiments, the compound having the structure of formula a-I or a salt thereof has NH (R a1 )-L a1 -C(R a2 )(R a3 )-L a2 -COOH structure or a salt thereof.
In some embodiments, the ring moiety, e.g., -Cy-, R (including those formed by R groups taken together), etc., is monocyclic. In some embodiments, the ring moiety is bicyclic or polycyclic. In some embodiments, the monocyclic ring is an optionally substituted 3 to 10 (3, 4, 5, 6, 7, 8, 9 or 10, 3 to 8, 3 to 7, 4 to 6, 5 to 6, etc.) membered saturated, partially unsaturated or aromatic ring having 0 to 5 heteroatoms. In some embodiments, each monocyclic ring unit of a bicyclic or polycyclic ring moiety is independently an optionally substituted 3 to 10 (3, 4, 5, 6, 7, 8, 9 or 10, 3 to 8, 3 to 7, 4 to 6, 5 to 6, etc.) membered saturated, partially unsaturated or aromatic ring having 0 to 5 heteroatoms.
In some embodiments, each heteroatom is independently selected from oxygen, nitrogen, sulfur, phosphorus, and silicon. In some embodiments, each heteroatom is independently selected from oxygen, nitrogen, and sulfur.
In some embodiments, L a1 Is a covalent bond. In some embodiments, the compound of formula A-1 has NH (R a1 )-C(R a2 )(R a3 )-L a2 -COOH structure.
In some embodiments, L a2 Is a covalent bond. In some embodiments, the compound of formula A-1 has NH (R a1 )-C(R a2 )(R a3 )-L a2 -COOH structure.
In some embodiments, L a1 Is a covalent bond, and L a2 Is a covalent bond. In some embodiments, the compound of formula A-1 has NH (R a1 )-C(R a2 )(R a3 ) -COOH structure.
In some embodiments, the amino acid is suitable for stapling. In some embodiments, the amino acid comprises a terminal olefin. Some such amino acids are exemplified herein (e.g., those described or used in peptides of multiple tables).
In some embodiments, the agent comprises a detectable moiety, which can be detected directly or indirectly. For example, in some embodiments, the detectable moiety is or comprises a fluorescent group. In some embodiments, the detectable moiety is or comprises a biotin moiety. In some embodiments, the detectable moiety is attached to the remainder of the agent at the amino acid residue, e.g., through a side chain, optionally through a linker (e.g., L as described herein). In some embodiments, the detectable moiety is-N 3 Which can be detected after a click chemistry reaction with a labeled reagent comprising an alkyne.
In some embodiments, the present disclosure provides a variety of compounds that are particularly useful as amino acids for many applications, such as for the preparation of peptides or other useful compounds.
In some embodiments, the compound (e.g., an amino acid or protected and/or activated form thereof) or salt thereof comprises 1) a first group that is an optionally protected amino group, 2) a second group that is an optionally protected and/or activated carboxyl group, and 3) a side chain (typically bonded to an atom between the first group and the second group ("side chain linking atom") that comprises an optionally protected and/or activated carboxyl group), and a) an optionally substituted ring (typically between the optionally protected and/or activated carboxyl group and the side chain linking atom of the side chain) or b) an amino group (typically between the optionally protected and/or activated carboxyl group and the side chain linking atom of the side chain). In some embodiments, provided compounds are optionally protected and/or activated amino acids or salts thereof, wherein the side chain of the amino acid comprises an optionally protected and/or activated carboxyl group and an optionally substituted ring or amino group, wherein the optionally substituted ring or amino group is between the backbone atoms to which the optionally protected and/or activated carboxyl group is attached to the side chain (e.g., the atoms between the amino group and the carboxyl group, both of which may be optionally and independently protected and/or activated (e.g., the alpha carbon atom in the amino acid)).
In some embodiments, the present disclosure provides compounds having the structure of formula PA or salts thereof:
N(R PA )(R a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)R PC ,
PA
wherein:
R PA is-H or an amino protecting group;
R a1 and R is a3 Each independently is-L a -R’；
R a2 is-L aa -C(O)R PS ；
L a 、L a1 And L a2 Each independently is L;
-C(O)R PS -COOH, optionally protected or activated;
-C(O)R PC -COOH, optionally protected or activated;
each L is independently a covalent bond or an optionally substituted divalent C 1 -C 25 Aliphatic or heteroaliphatic groups having from 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R')-, -C (O) S-or-C (O) O-;
each-Cy-is independently an optionally substituted divalent 3-to 30-membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms;
each R' is independently-R, -C (O) R, -CO 2 R or-SO 2 R is R; and is also provided with
Each R is independently-H or an optionally substituted group selected from: c (C) 1-30 Aliphatic, C having 1 to 10 heteroatoms 1-30 Heteroaliphatic, C 6-30 Aryl, C 6-30 Arylaliphatic, C having 1 to 10 heteroatoms 6-30 Aryl heteroaliphatics, 5 to 30 membered heteroaryl groups having 1 to 10 heteroatoms and 3 to 30 membered heterocyclyl groups having 1 to 10 heteroatoms, or
The two R groups optionally and independently together form a covalent bond, or:
Two or more R groups on the same atom optionally and independently taken together with the atom form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the atom; or alternatively
Two or more R groups on two or more atoms optionally and independently together with their intervening atoms form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the intervening atoms.
In some embodiments, a compound having the structure of formula PA (e.g., an amino acid, such as those of formulas a-I or protected/activated forms thereof) or a salt thereof:
N(R PA )(R a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)R PC ,
PA
wherein:
R PA is-H or an amino protecting group;
R a1 and R is a3 Each independently is-L a -R’；
R a2 is-L aa -C(O)R PS Wherein L is aa Is L, and L aa comprising-N (R') -or-Cy-;
L a1 and L a2 Each independently is L;
-C(O)R PS -COOH, optionally protected or activated;
-C(O)R PC -COOH, optionally protected or activated;
each L is independently a covalent bond or an optionally substituted divalent C 1 -C 25 Aliphatic or heteroaliphatic groups having from 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R')-, -C (O) S-or-C (O) O-;
each-Cy-is independently an optionally substituted divalent 3-to 30-membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms;
each R' is independently-R, -C (O) R, -CO 2 R or-SO 2 R is R; and is also provided with
Each R is independently-H or an optionally substituted group selected from: c (C) 1-30 Aliphatic, C having 1 to 10 heteroatoms 1-30 Heteroaliphatic, C 6-30 Aryl, C 6-30 Arylaliphatic, C having 1 to 10 heteroatoms 6-30 Aryl heteroaliphatics, 5 to 30 membered heteroaryl groups having 1 to 10 heteroatoms and 3 to 30 membered heterocyclyl groups having 1 to 10 heteroatoms, or
The two R groups optionally and independently together form a covalent bond, or:
two or more R groups on the same atom optionally and independently taken together with the atom form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the atom; or alternatively
Two or more R groups on two or more atoms optionally and independently together with their intervening atoms form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the intervening atoms.
In some embodiments, L a1 Is a covalent bond. In some embodiments, L a1 Not a covalent bond.
In some embodiments, L a2 Is a covalent bond. In some embodiments, L a2 Not a covalent bond.
In some embodiments, R a2 is-L aa -C(O)R PS Wherein L is aa Is an optionally substituted divalent C 1 -C 25 Aliphatic or heteroaliphatic groups having from 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R')-, -C (O) S-or-C (O) O-, wherein at least one methylene unit is replaced by-Cy-.
As used herein, in some embodiments, -Cy-is an optionally substituted divalent 3 to 10 (e.g., 3, 4, 5, 6, 7, 8, 9, or 10) membered monocyclic cycloaliphatic group. In some embodiments, -Cy-is an optionally substituted 3-to 10-membered (e.g., 3, 4, 5, 6, 7, 8, 9, or 10) monocyclic cycloalkyl ring. In some embodiments, -Cy-is an optionally substituted 3 to 10 (e.g., 3, 4, 5, 6, 7, 8, 9, or 10) membered monocyclic heteroalicyclic ring having 1 to 5 heteroatoms. In some embodiments, -Cy-is an optionally substituted 3 to 10 (e.g., 3, 4, 5, 6, 7, 8, 9, or 10) membered monocyclic heteroalkyl ring having 1 to 5 heteroatoms. In some embodiments, -Cy-is an optionally substituted divalent 5-15 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15) membered bicyclic or polycyclic cycloaliphatic group. In some embodiments, -Cy-is an optionally substituted divalent 5-15 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15) membered bicyclic or polycyclic cycloalkyl. In some embodiments, -Cy-is an optionally substituted 5 to 15 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15) membered bicyclic or polycyclic heteroalicyclic having 1 to 5 heteroatoms. In some embodiments, -Cy-is an optionally substituted 5 to 15 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15) membered bicyclic or polycyclic heterocyclyl ring having 1 to 5 heteroatoms. In some embodiments, the cycloaliphatic, cycloalkyl, heteroaliphatic, or heteroalkyl ring is 3-membered. In some embodiments, it is 4-membered. In some embodiments, it is 5-membered. In some embodiments, it is 6 membered. In some embodiments, it is 7-membered. In some embodiments, it is 8-membered. In some embodiments, it is 9-membered. In some embodiments, it is 10 membered. In some embodiments, it is 11 membered. In some embodiments, it is 12 membered. In some embodiments, -Cy-is optionally substituted phenylene. In some embodiments, -Cy-is an optionally substituted divalent 10 membered bicyclic aryl ring. In some embodiments, -Cy-is an optionally substituted 5 membered heteroaryl ring having 1 to 4 heteroatoms. In some embodiments, -Cy-is an optionally substituted 6 membered heteroaryl ring having 1 to 4 heteroatoms. In some embodiments, -Cy-is an optionally substituted 9 membered bicyclic heteroaryl ring having 1 to 5 heteroatoms. In some embodiments, -Cy-is an optionally substituted 10 membered bicyclic heteroaryl ring having 1 to 5 heteroatoms. In some embodiments, the heteroaliphatic, heterocyclic, or heteroaryl ring contains no more than 1 heteroatom. In some embodiments, each heteroatom is independently selected from nitrogen, oxygen, and sulfur.
In some embodiments, -Cy-is an optionally substituted 4-to 7-membered ring having 0 to 3 heteroatoms. In some embodiments, -Cy-is an optionally substituted 6 membered aryl ring. In some embodiments, the aryl ring is substituted. In some embodiments, it is substituted with one or more halogens. In some embodiments, it is substituted with one or more-F. In some embodiments, it is not substituted. In some embodiments, it is optionally substitutedIn some embodiments, it is +.>In some embodiments, it is optionally substituted +.>In some embodiments, it is +.>In some embodiments, it is optionally substituted +.>In some embodiments, it is +.>In some embodiments, -Cy-is an optionally substituted 5 membered heteroaryl ring having 1 to 3 heteroatoms. In some embodiments, the heteroatom is nitrogen. In some embodiments, the heteroatom is oxygen. In some embodiments, the heteroatom is sulfur. In some embodiments, -Cy-is optionally substituted +.>In some embodiments, -Cy-is +.>
In some embodiments, L aa is-L am1 -Cy-L am2 -, wherein L am1 And L am2 Each independently is L am Wherein each L am Independently a covalent bond or an optionally substituted divalent C 1 -C 10 An aliphatic group, wherein one or more methylene units of the aliphatic group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-.
In some embodiments, L aa comprises-Cy-. In some embodiments, L aa is-L am1 -Cy-L am2 -, wherein L am1 And L am2 Each independently is L am Wherein each L am Independently a covalent bond or an optionally substituted divalent C 1 -C 10 An aliphatic group, wherein one or more methylene units of the aliphatic group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, -L am2 -and-C (O) R PS And (5) bonding. In some embodiments, L am2 Is a covalent bond. In some embodiments, -Cy-is an optionally substituted 4-to 7-membered ring having 0 to 3 heteroatoms. In some embodiments, -Cy-is an optionally substituted 5-to 7-membered ring having 0 to 3 heteroatoms. In some embodiments, -Cy-is an optionally substituted 6-to 7-membered ring having 0 to 3 heteroatoms. In some embodiments, -Cy-is an optionally substituted 4 membered ring having 0 to 1 heteroatoms. In some embodiments, -Cy-is an optionally substituted 5 membered ring having 0 to 2 heteroatoms. In some embodiments, -Cy-is an optionally substituted 6 membered ring having 0 to 2 heteroatoms. In some embodiments, -C y-is an optionally substituted 7 membered ring having 0 to 3 heteroatoms.
In some embodiments, R a2 is-L aa -C(O)R PS Wherein L is aa Is an optionally substituted divalent C 1 -C 25 Aliphatic or heteroaliphatic groups having from 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R ') -, C (O) S-, or-C (O) O-, wherein at least one methylene unit is replaced by-N (R') -.
In some embodiments, L aa comprising-N (R') -. In some embodiments, L aa is-L am1 -(NR’)-L am2 -, wherein L am1 And L am2 Each independently is L am Wherein each L am Independently a covalent bond or an optionally substituted divalent C 1 -C 10 An aliphatic group, wherein one or more methylene units of the aliphatic group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, -L am2 -and-C (O) R PS And (5) bonding. In some embodiments, L am1 Is optionally substituted C 1-4 An alkylene group. In some embodiments, L am1 Is optionally substituted- (CH) 2 ) m-, wherein m is 1, 2, 3 or 4. In some embodiments, L am1 is-CH 2 -. In some embodiments, L am2 Is an optionally substituted straight chain C 1-2 An alkylene group. In some embodiments, L am2 Is- [ C (R') 2 ]n, where n is 1 or 2. In some embodiments, L am2 Is- [ CHR ]']n, where n is 1 or 2. In some embodiments, each R' is independently-H or optionally takenSubstituted C 1-6 An alkyl group. In some embodiments, L am2 Is optionally substituted-CH 2 -. In some embodiments, L am2 is-CH 2 -. In some embodiments, R' is-R NR Wherein R is NR Is R. In some embodiments, R' is-CH 2 -R NR Wherein R is NR Is R. In some embodiments, R 'in-N (R') -is-C (O) R NR Wherein R is NR Is R. In some embodiments, R 'in-N (R') -is-SO 2 R NR Wherein R is NR Is R. In some embodiments, R is optionally substituted C 1-6 Aliphatic or heteroaliphatic having 1 to 4 heteroatoms. In some embodiments, R NR Is C 1-7 Alkyl or heteroalkyl having 1 to 4 heteroatoms, optionally substituted with one or more C's independently selected from halogen, having 0 to 4 heteroatoms 5-6 An aromatic ring and optionally substituted 3-to 10-membered cycloalkyl or a heteroalkyl ring having 1 to 4 heteroatoms. In some embodiments, R is-CF 3 . In some embodiments, L am2 is-C (R') 2 -or comprises-C (R') 2 -wherein R ' in the R ' group and-N (R ') -together with its intervening atoms form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the intervening atoms.
In some embodiments, L aa is-L am1 -N(R’)-L am2 -, wherein L am1 And L am2 Each independently is L am Wherein each L am Independently a covalent bond or an optionally substituted divalent C 1 -C 10 An aliphatic group, wherein one or more methylene units of the aliphatic group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-.
In some embodimentsIn this case, -N (R'), -is bonded to two carbon atoms which do not form any double bonds with the hetero atom. In some embodiments, -N (R') -is bonded to two sp3 atoms. In some embodiments, -N (R') -is bonded to two sp3 carbon atoms. In some embodiments, -N (R'), -and two-CH 2 Bonding, each-CH 2 Independently and optionally substituted with one or two monovalent substituents. In some embodiments, -N (R'), -and two-CH 2 -bonding.
In some embodiments, L aa comprising-N (R') -. In some embodiments, R 'of-N (R') -is-R NR Wherein R is NR Is R. In some embodiments, R 'of-N (R') -is-CH 2 -R NR Wherein R is NR Is R, and-CH 2 -is optionally substituted. In some embodiments, R 'in-N (R') -is-C (O) R NR Wherein R is NR Is R. In some embodiments, R 'in-N (R') -is-SO 2 R NR Wherein R is NR Is R. In some embodiments, -N (R'), -is-N (Et) -. In some embodiments, -N (R'), -is-N (CH) 2 CF 3 ) -. In some embodiments, R' is optionally substituted C 1-6 Aliphatic or heteroaliphatic having 1 to 4 heteroatoms. In some embodiments, R' is C 1-7 An alkyl group or a heteroalkyl group having 1 to 4 heteroatoms, wherein the alkyl or heteroalkyl group is optionally substituted with one or more C's independently selected from halogen, having 0 to 4 heteroatoms 5-6 An aromatic ring and optionally substituted 3-to 10-membered cycloalkyl or a heteroalkyl ring having 1 to 4 heteroatoms. In some embodiments, R NR is-CF 3 。
In some embodiments, R 'in-N (R') -is R, R a3 Is R and the two R groups together with the intervening atoms form an optionally substituted 3 to 10 membered ring having 0 to 5 heteroatoms in addition to the intervening atoms. In some embodiments, the ring formed is 3-membered. In some embodiments, the ring formed is 4-membered. In some embodiments, the ring formed is 5-membered.In some embodiments, the ring formed is 6 membered. In some embodiments, the ring formed is 7 membered. In some embodiments, the ring formed is monocyclic. In some embodiments, the rings formed are bicyclic or polycyclic. In some embodiments, the formed ring is saturated. In some embodiments, the rings formed are partially unsaturated.
In some embodiments, L am1 Is a covalent bond. In some embodiments, L am1 Not a covalent bond. In some embodiments, L am1 Is optionally substituted C 1-4 An alkylene group. In some embodiments, L am1 Is optionally substituted- (CH) 2 ) m-, wherein m is 1, 2, 3 or 4. In some embodiments, L am1 Is optionally substituted-CH 2 -. In some embodiments, L am1 is-CH 2 -。
In some embodiments, L am2 and-C (O) R PS And (5) bonding.
In some embodiments, L am2 Is a covalent bond. In some embodiments, when L am2 at-Cy-and-C (O) R PS Which are covalent bonds when in between. In some embodiments, L am2 Not a covalent bond. In some embodiments, L am2 Is optionally substituted C 1-4 An alkylene group. In some embodiments, L am2 Is optionally substituted- (CH) 2 ) m-, wherein m is 1, 2, 3 or 4. In some embodiments, L am2 Is an optionally substituted straight chain C 1-2 An alkylene group. In some embodiments, L am2 Is- [ C (R') 2 ]n, where n is 1 or 2. In some embodiments, L am2 Is- [ CHR ]']n, where n is 1 or 2. In some embodiments, each R' is independently-H or optionally substituted C 1-6 An alkyl group. In some embodiments, L am2 Is optionally substituted-CH 2 -. In some embodiments, L am2 is-CH 2 -. In some embodiments, L am2 Is optionally substituted-CH 2 -CH 2 -. In some embodiments, L am2 Is-CH 2 -C(CH 3 ) 2 -。
In some embodiments, L am2 is-C (R') 2 -or comprises-C (R') 2 -, wherein R' is a radical and L aa R 'in (R') -together with the intervening atoms thereof form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the intervening atoms.
In some embodiments, R a2 is-L aa -C(O)R PS Wherein L is aa Is L, as described herein. In some embodiments, L aa Is L am2 As described herein. In some embodiments, L aa Is an optionally substituted branched or straight chain C 1-10 A hydrocarbon chain. In some embodiments, L aa Is optionally substituted C 1-10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) alkylene. In some embodiments, L aa Is optionally substituted-CH 2 -CH 2 -. In some embodiments, L aa is-CH 2 -CH 2 -. In some embodiments, L aa Is optionally substituted-CH 2 -. In some embodiments, L aa is-CH 2 -。
In some embodiments, L a Is L aa As described herein.
In some embodiments, L aa Is L a As described herein.
As described above, each L is independently a covalent bond or an optionally substituted divalent C 1 -C 25 Aliphatic or heteroaliphatic groups having from 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-.
In some embodiments, L is a covalent bond.
In some embodiments, L (or L a 、L aa 、L a1 、L a2 、L s1 、L s2 、L s3 Or another variable or moiety that may be L, or a linker moiety) is an optionally substituted divalent C 1 -C 25 、C 1 -C 20 、C 1 -C 15 、C 1 -C 10 、C 1 -C 9 、C 1 -C 8 、C 1 -C 7 、C 1 -C 6 、C 1 -C 5 、C 1 -C 4 、C 1 -C 3 、C 1 -C 2 Or C 1 、C 2 、C 3 、C 4 、C 5 、C 6 、C 7 、C 8 、C 9 、C 10 、C 11 、C 12 、C 13 、C 14 、C 15 、C 16 、C 17 、C 18 、C 19 Or C 20 Aliphatic, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-.
In some embodiments, L, L a 、L aa 、L a1 、L a2 、L s1 、L s2 、L s3 L ", or another variable or moiety that may be L, or the linker moiety is an optionally substituted divalent C 1 -C 25 、C 1 -C 20 、C 1 -C 15 、C 1 -C 10 、C 1 -C 9 、C 1 -C 8 、C 1 -C 7 、C 1 -C 6 、C 1 -C 5 、C 1 -C 4 、C 1 -C 3 、C 1 -C 2 Or C 1 、C 2 、C 3 、C 4 、C 5 、C 6 、C 7 、C 8 、C 9 、C 10 、C 11 、C 12 、C 13 、C 14 、C 15 、C 16 、C 17 、C 18 、C 19 Or C 20 Aliphatic, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, it is an optionally substituted divalent C 1 -C 10 、C 1 -C 9 、C 1 -C 8 、C 1 -C 7 、C 1 -C 6 、C 1 -C 5 、C 1 -C 4 、C 1 -C 3 、C 1 -C 2 Or C 1 、C 2 、C 3 、C 4 、C 5 、C 6 、C 7 、C 8 、C 9 Or C 10 Aliphatic, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, it is an optionally substituted divalent C 2 Aliphatic, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, it is an optionally substituted divalent C 3 Aliphatic, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, it is an optionally substituted divalent C 4 Aliphatic, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, it is an optionally substituted divalent C 5 Aliphatic, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, it is an optionally substituted divalent C 6 Aliphatic, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, the divalent aliphatic group is saturated. In some embodiments, the divalent aliphatic group is linear. In some embodiments, the divalent aliphatic group is branched. In some embodiments, it is an optionally substituted divalent straight chain saturated C 6 Aliphatic, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, each substitution (if any) is independently replaced with-Cy-, -O-, -S-S-, -N (R '), -C (O) -, -C (S) -, -C (NR '), -C (O) N (R '), -N @R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments, each substitution (if any) is independently replaced with-Cy-, -O-, -S-, -N (R '), -C (O) -, -C (S) -, -C (NR'), -C (O) N (R '), -N (R') C (O) O-, -S (O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments of the present invention, in some embodiments, each substitution, if any, is independently made with-O-, -S-, -N (R'), -C (O) -, -S (O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-. In some embodiments of the present invention, in some embodiments, each substitution, if any, is independently replaced with-O-, -S-, -N (R') -or-C (O) -. In some embodiments, L, L a 、L aa 、L a1 、L a2 、L s1 、L s2 、L s3 L ", or another variable or moiety that may be L, or the linker moiety is an optionally substituted divalent C 1 -C 6 Straight chain saturated aliphatic wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R') -or-C (O) -. In some embodiments, it is an optionally substituted divalent C 1 -C 5 Straight chain saturated aliphatic wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R') -or-C (O) -. In some embodiments, it is an optionally substituted divalent C 1 -C 4 Straight chain saturated aliphatic wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R') -or-C (O) -. In some embodiments, it is an optionally substituted divalent C 1 -C 3 Straight chain saturated aliphatic wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R') -or-C (O) -. In some embodiments, it is an optionally substituted divalent C 1 -C 2 Straight chain saturated aliphatic wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R') -or-C (O) -. In some embodiments, it is a divalent C 1 -C 6 Straight-chain saturated aliphatic wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R') -or-C (O) -substitution. In some embodiments, it is a divalent C 1 -C 5 Straight chain saturated aliphatic wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R') -or-C (O) -. In some embodiments, it is a divalent C 1 -C 4 Straight chain saturated aliphatic wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R') -or-C (O) -. In some embodiments, it is a divalent C 1 -C 3 Straight chain saturated aliphatic wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R') -or-C (O) -. In some embodiments, it is a divalent C 1 -C 2 Straight chain saturated aliphatic wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R') -or-C (O) -. In some embodiments, no substitution of methylene units is present. In some embodiments, there is an alternative. In some embodiments, there are two alternatives. In some embodiments, there are three alternatives. In some embodiments, there are four or more substitutions. In some embodiments, R 'in each moiety (e.g., -N (R') -) used to replace a methylene unit as described herein is hydrogen or optionally substituted C 1-6 Aliphatic or phenyl. In some embodiments, R' in each such moiety is hydrogen or optionally substituted C 1-6 An alkyl group. In some embodiments, R' in each such moiety is hydrogen or C 1-6 An alkyl group. In some embodiments, each-Cy-is an optionally substituted divalent ring selected from 3 to 10, 3 to 9, 3 to 8, 3 to 7, 5 to 10, 5 to 9, 5 to 8, 5 to 7, 5 to 6, or 3, 4, 5, 6, 7, 8, 9, or 10 membered cycloaliphatic and heterocyclylene having 1 to 3 heteroatoms, phenylene, and 5 to 6 membered heteroarylene having 1 to 3 heteroatoms. In some embodiments, -Cy-is an optionally substituted divalent 3 to 10, 3 to 9, 3 to 8, 3 to 7, 5 to 10, 5 to 9, 5 to 8, 5 to 7, 5 to 6, or 3, 4, 5, 6, 7, 8, 9, or 10 membered cycloaliphatic. In some embodiments, -Cy-is an optionally substituted 3 to 10, 3 to 9, 3 to 8, 3 to 7, 5 to 10, 5 to 9, 5 to 8, optionally substituted with 1 to 3 heteroatoms,5 to 7, 5 to 6, or 3, 4, 5, 6, 7, 8, 9, or 10 membered heterocyclylene. In some embodiments, -Cy-is an optionally substituted 3 to 10, 3 to 9, 3 to 8, 3 to 7, 5 to 10, 5 to 9, 5 to 8, 5 to 7, 5 to 6, or 3, 4, 5, 6, 7, 8, 9, or 10 membered heterocyclyl. In some embodiments, -Cy-is optionally substituted phenylene. In some embodiments, -Cy-is phenylene. In some embodiments, -Cy-is an optionally substituted 5-to 6-membered heteroarylene having 1 to 3 heteroatoms. In some embodiments, -Cy-is an optionally substituted 5-to 6-membered heteroarylene having 1 heteroatom. In some embodiments, the heteroatom is nitrogen. In some embodiments, the heteroatom is oxygen. In some embodiments, the heteroatom is sulfur. In some embodiments, L, L a 、L aa 、L a1 、L a2 、L s1 、L s2 、L s3 L ", or another variable or moiety which may be L, or the linker moiety is optionally substituted- (CH) 2 ) n-. In some embodiments, it is- (CH) 2 ) n-. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 6. In some embodiments, n is 7. In some embodiments, n is 8. In some embodiments, n is 9. In some embodiments, n is 10.
In some embodiments, L, L a 、L aa 、L a1 、L a2 、L s1 、L s2 、L s3 Another variable or moiety, or linker moiety, which may be L is an optionally substituted divalent heteroaliphatic group having 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-Cy-、-O-、-S-、-S-S-、-N(R’)-、-C(O)-、-C(S)-、-C(NR’)-、-C(O)N(R’)-、-N(R’)C(O)N(R’)-、-N(R’)C(O)O-、-S(O)-、-S(O) 2 -、-S(O) 2 N (R') - (C (O) S) -or-C (O) O-.
Those skilled in the art understand that for a linker moiety that may be L or L "(e.g., L aa 、L s1 、L s2 、L s3 、L s 、L a 、L a1 、L a2 、L RN Etc.) may also be used for another group which may be L or L "to the extent that such an embodiment falls within the definition of L or L".
As described above, each R' is independently-R, -C (O) R, -CO 2 R or-SO 2 R is defined as the formula. In some embodiments, R' is-L a -R. In some embodiments, R' is R. In some embodiments, R' is-C (O) R. In some embodiments, R' is-CO 2 R is defined as the formula. In some embodiments, R' is-SO 2 R is defined as the formula. In some embodiments, R' is-H.
As described above, each R is independently-H, or an optionally substituted group selected from: c (C) 1-30 Aliphatic, C having 1 to 10 heteroatoms 1-30 Heteroaliphatic, C 6-30 Aryl, C 6-30 Arylaliphatic, C having 1 to 10 heteroatoms 6-30 Aryl heteroaliphatics, 5 to 30 membered heteroaryl groups having 1 to 10 heteroatoms and 3 to 30 membered heterocyclyl groups having 1 to 10 heteroatoms, or
The two R groups optionally and independently taken together form a covalent bond, or
Two or more R groups on the same atom optionally and independently taken together with the atom form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the atom; or alternatively
Two or more R groups on two or more atoms optionally and independently together with their intervening atoms form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the intervening atoms.
In some embodiments, R is-H, as described herein. In some embodiments, R is not-H. In some embodiments of the present invention, in some embodiments,r is optionally substituted C 1-10 Aliphatic series. In some embodiments, R is optionally substituted C 1-10 An alkyl group. In some embodiments, R is methyl. In some embodiments, R is ethyl. In some embodiments, R is isopropyl. In some embodiments, R is-CF 3 . In some embodiments, R is-CH 2 CF 3 . In some embodiments, R is butyl. In some embodiments, R is tert-butyl. In some embodiments, R is optionally substituted C 3-10 Cycloaliphatic. In some embodiments, R is optionally substituted C 3-10 Cycloalkyl groups. In some embodiments, R is optionally substituted cyclopropyl. In some embodiments, R is optionally substituted cyclobutyl. In some embodiments, R is optionally substituted cyclopentyl. In some embodiments, R is optionally substituted cyclohexyl. In some embodiments, R is optionally substituted phenyl. In some embodiments, R is phenyl. In some embodiments, R is an optionally substituted 5 membered heteroaryl having 1 to 3 heteroatoms. In some embodiments, R is an optionally substituted 5 membered heteroaryl having 1 heteroatom. In some embodiments, R is an optionally substituted 6 membered heteroaryl having 1 to 3 heteroatoms. In some embodiments, R is an optionally substituted 6 membered heteroaryl having 1 heteroatom. In some embodiments, R is an optionally substituted bicyclic 8-to 10-membered aromatic ring having 0-5 heteroatoms. In some embodiments, R is an optionally substituted bicyclic 9-membered aromatic ring having 1 to 5 heteroatoms. In some embodiments, R is an optionally substituted bicyclic 10 membered aromatic ring having 1 to 5 heteroatoms. In some embodiments, R is an optionally substituted bicyclic 9-membered aromatic ring having 1 heteroatom. In some embodiments, R is an optionally substituted bicyclic 10 membered aromatic ring having 1 heteroatom. In some embodiments, R is an optionally substituted bicyclic 10-membered aromatic ring having no heteroatoms. In some embodiments, R is an optionally substituted 3-to 10-membered heterocyclyl having 1 to 5 heteroatoms. In some embodiments, R is optionally substituted having 1 to 5 5-to 14-membered bicyclic heterocyclyl groups of heteroatoms.
In some embodiments, two R groups (or two groups that may be R, e.g., two are each independently selected from R', R a1 、R a2 、R a3 、R a5 、R RN Etc.) together with intervening atoms to form an optionally substituted 3-to 30-membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the atoms. In some embodiments, the ring formed is substituted. In some embodiments, the ring formed is unsubstituted. In some embodiments, the formed ring is 3 to 30, 3 to 20, 3 to 15, 3 to 10, 3 to 9, 3 to 8, 3 to 7, 3 to 6, 4 to 10, 4 to 9, 4 to 8, 4 to 7, 4 to 6, 5 to 10, 5 to 9, 5 to 8, 5 to 7, 5 to 6, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 membered. In some embodiments, the ring formed is 3 to 10 membered. In some embodiments, the ring formed is 3 to 7 membered. In some embodiments, the ring formed is 4 to 10 membered. In some embodiments, the ring formed is 4 to 7 membered. In some embodiments, the ring formed is 5 to 10 membered. In some embodiments, the ring formed is 5 to 7 membered. In some embodiments, the ring formed is 3-membered. In some embodiments, the ring formed is 4-membered. In some embodiments, the ring formed is 5-membered. In some embodiments, the ring formed is 6 membered. In some embodiments, the ring formed is 7 membered. In some embodiments, the ring formed is 8-membered. In some embodiments, the ring formed is 9-membered. In some embodiments, the ring formed is 10 membered. In some embodiments, the ring formed is monocyclic. In some embodiments, the ring formed is bicyclic. In some embodiments, the rings formed are polycyclic. In some embodiments, the formed ring has no heteroatoms other than intervening atoms. In some embodiments, the formed ring has 1 to 10, for example 1 to 5, 1 to 3, or 1, 2, 3, 4, in addition to intervening atoms, 5. 6, 7, 8, 9 or 10 heteroatoms. In some embodiments, the formed ring is saturated. In some embodiments, the rings formed are partially unsaturated. In some embodiments, the formed ring comprises one or more aromatic rings. In some embodiments, the rings formed are bicyclic or polycyclic, and each monocyclic unit is independently 3 to 10 membered saturated, partially unsaturated, or aromatic, and has 0 to 5 heteroatoms. In some embodiments, each heteroatom is independently selected from nitrogen, oxygen, and sulfur.
In some embodiments, may be R, e.g., R', R a1 、R a2 、R a3 、R a5 、R RN And the like are R, as described herein. Those skilled in the art will appreciate that the embodiments described for one group that may be R may also be used for another group that may be R to the extent that such embodiments fall within the definition of R.
In some embodiments, the present disclosure provides a compound having the structure:
wherein:
m and n are each independently 1, 2, 3 or 4;
L RN is L;
R RN is R;
R a5 is R'; and is also provided with
Each other variable is independently as described herein.
In some embodiments, m is 1. In some embodiments, m is 2. In some embodiments, m is 3. In some embodiments, m is 4.
In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4.
In some embodiments, L RN is-CH 2 -, -CO-or-SO 2 -. In some embodiments, L RN is-CH 2 -. In some embodiments, L RN is-CO-. In some embodiments, L RN is-SO 2 -. In some embodiments, L RN Is an optionally substituted divalent C 1-4 An alkylene group. In some embodiments, L RN Is an optionally substituted divalent straight chain C 1-4 An alkylene group. In some embodiments, L RN is-CH 2 -CH 2 -. In some embodiments, L RN is-CH 2 -CH 2 -CH 2 -. In some embodiments, L RN is-C (CH) 3 )-。
In some embodiments, R RN Is R, as described herein. In some embodiments, R RN Is C 1-7 An alkyl group or a heteroalkyl group having 1 to 4 heteroatoms, wherein the alkyl or heteroalkyl group is optionally substituted with one or more C's independently selected from halogen, having 0 to 4 heteroatoms 5-6 An aromatic ring and optionally substituted 3-to 10-membered cycloalkyl or a heteroalkyl ring having 1 to 4 heteroatoms.
In some embodiments, R (e.g., R RN R', etc.) is an optionally substituted aliphatic, e.g., C 1-10 Aliphatic series. In some embodiments, R is optionally substituted alkyl, e.g., C 1-10 An alkyl group. In some embodiments, R is optionally substituted cycloalkyl, e.g., C 1-10 Cycloalkyl groups. In some embodiments, R is optionally substituted aryl. In some embodiments, R is optionally substituted heterocyclyl. In some embodiments, R is optionally substituted heteroaryl. In some embodiments, is methyl. In some embodiments, R is-CF 3 . In some embodiments, R is ethyl. In some embodiments, R isIn some embodiments, R is phenyl. In some embodiments, R is pentafluorophenyl. In some embodiments, R is pyridinyl.
In some embodiments, one or more R a5 Independently is-H. In some embodiments, one or more R a5 Independently optionally substituted C 1-6 An alkyl group. In some embodiments, each R a5 is-H.
In some embodiments, -L RN -R RN Is R and is with R a5 And intervening atoms together form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to intervening atoms.
As described in this disclosure, the various rings, including those in multiple moieties (e.g., R or various groups that can be R, various divalent rings, such as those in-Cy) and those formed by two entities (e.g., two groups that can be R) taken together with an intervening form of R, can have multiple sizes, e.g., 3 to 30. In some embodiments, the ring is 3 to 30 membered. In some embodiments, the ring is 3 to 20 membered. In some embodiments, the ring is 3 to 10 membered. In some embodiments, the ring is, for example, 3, 4, 5, 6, 7, 8, 9, or 10 membered. In some embodiments, the ring is 3-membered. In some embodiments, the ring is 4 membered. In some embodiments, the ring is 5 membered. In some embodiments, the ring is 6 membered. In some embodiments, the ring is 7 membered. In some embodiments, the ring is 8 membered. In some embodiments, the ring is 9 membered. In some embodiments, the ring is 10 membered. In some embodiments, the ring is substituted (in addition to the latent groups already shown in the formula). In some embodiments, the ring is unsubstituted. In some embodiments, the ring is saturated. In some embodiments, the ring is partially unsaturated. In some embodiments, the ring is aromatic. In some embodiments, the ring comprises one or more, e.g., 1 to 5 heteroatoms. In some embodiments, one or more heteroatoms is oxygen. In some embodiments, one or more heteroatoms is nitrogen. In some embodiments, one or more heteroatoms is sulfur. In some embodiments, the ring is a cycloaliphatic ring, such as a cycloalkyl ring. In some embodiments, the ring is a heterocyclic aliphatic ring, such as a heterocycloalkyl ring. In some embodiments, the ring is an aryl ring. In some embodiments, the ring is a heteroaryl ring. In some embodiments, the ring is a heteroaryl ring. In some embodiments, the ring is monocyclic. In some embodiments, the ring is bicyclic or polycyclic. In some embodiments, each monocyclic unit in the ring is independently an optionally substituted 3-to 10-membered (e.g., 3, 4, 5, 6, 7, 8, 9, or 10-membered) saturated, partially unsaturated, or aromatic ring having 0 to 5 heteroatoms.
In some embodiments, the heteroatom is selected from nitrogen, oxygen, sulfur, silicon, and phosphorus, as described herein. In some embodiments, as described herein, the heteroatom is selected from nitrogen, oxygen, and sulfur.
In some embodiments, R a1 is-H. In some embodiments, R a1 Is optionally substituted C 1-6 An alkyl group. In some embodiments, R a1 With another group (e.g. R a3 ) And intervening atoms therebetween, together form an optionally substituted ring as described herein.
In some embodiments, -C (O) R PC Is a protected carboxylic acid group. In some embodiments, -C (O) R PC Is an activated carboxylic acid group. Those of skill in the art will appreciate that a variety of groups may be used to protect/activate carboxyl groups, including a variety of groups useful in peptide synthesis, and may be used in accordance with the present disclosure. In some embodiments, -C (O) R PC Is an ester. In some embodiments, -C (O) R PC Is an activated ester for synthesis. In some embodiments, -C (O) R PC is-C (O) OR'. In some embodiments, R' is R. In some embodiments, R' is optionally substituted C 1-10 Aliphatic series. In some embodiments, R' is optionally substituted phenyl. In some embodiments, R' is pentafluorophenyl. In some embodiments, R' is
In some embodiments, -C (O) R PC is-COOH.
In some embodiments, -C (O) R PS Is a protected carboxylic acid group. In some embodiments, if-C (O) R PS To react with another moiety, it is an activated carboxylic acid group. Those of skill in the art will appreciate that a variety of groups may be used to protect/activate carboxyl groups, including a variety of groups useful in peptide synthesis, and may be used in accordance with the present disclosure. In some embodiments, -C (O) R PS Is an ester. In some embodiments, -C (O) R PS Is an ester. In some embodiments, -C (O) R PS is-C (O) OR'. In some embodiments, R' is R. In some embodiments, R is optionally substituted C 1-10 Aliphatic series. In some embodiments, R is optionally substituted phenyl. In some embodiments, R is optionally substituted t-Bu. In some embodiments, R is t-Bu. In some embodiments, R is benzyl. In some embodiments, R is allyl. In some embodiments, -C (O) R PS Is a protected carboxylic acid group compatible with peptide synthesis (e.g., fmoc-based peptide synthesis). In some embodiments, -C (O) R PS Is with-C (O) R PC And R is PA Orthogonal protected carboxylic acid groups, and at-C (O) R PC And/or N (R) PA )(R a1 ) Is protected, deprotected, and/or reacted (e.g., in peptide synthesis, such as Fmoc-based peptide synthesis). In some embodiments, -C (O) R PS Is deprotected later during synthesis, for example, after the peptide backbone is constructed or mostly constructed so that unprotected side chains-COOH do not affect synthesis.
In some embodiments, -C (O) R PS is-COOH.
As described above, R PA is-H or an amino protecting group. In some embodiments, R PA is-H. In some embodiments, R PA Is an amino protecting group. In some embodiments, R PA Is suitable for amino protection of peptide synthesisAnd (3) a protecting base. In some embodiments, R PA is-C (O) -O-R, wherein R is optionally substitutedIn some embodiments, R PA is-Fmoc. In some embodiments, R PA is-Cbz. In some embodiments, R PA is-Boc.
In some embodiments, R PS Is with R PA Orthogonal protecting groups. In some embodiments, R PS Is with R PC Orthogonal protecting groups. In some embodiments, R PS Is compatible with peptide synthesis. In some embodiments, R PS Is optionally substituted C 1-6 Aliphatic series. In some embodiments, R PS Is tert-butyl.
In some embodiments, R PS is-S-L-R', wherein each variable is independently as described herein. In some embodiments, L is an optionally substituted-CH 2 -. In some embodiments, L is-CH 2 -. In some embodiments, R PS is-S-CH 2 -R ', wherein R' is as described herein. In some embodiments, R' is R, as described herein. In some embodiments, R is optionally substituted C 6-30 Aryl groups. In some embodiments, R is optionally substituted C 6-10 Aryl groups. In some embodiments, R is optionally substituted phenyl. In some embodiments, R is phenyl. In some embodiments, R is substituted phenyl, wherein one or more substituents are independently alkoxy. In some embodiments, R is 2,4, 6-trimethoxyphenyl. In some embodiments, R is an optionally substituted 5-to 30-membered heteroaryl having 1 to 10 heteroatoms. In some embodiments, R is an optionally substituted 5-to 10-membered heteroaryl having 1-4 heteroatoms. In some embodiments, R is an optionally substituted 5 membered heteroaryl having 1 to 4 heteroatoms. In some embodiments, R PS is-S-CH 2 -Cy-R', wherein-CH 2 -is optionally substituted, and-Cy-is as described herein. In some embodiments, R PS is-S-CH 2 -Cy-O-R', wherein-CH 2 -is optionally substituted, and-Cy-is as described herein. In some embodiments, -Cy-is an optionally substituted aromatic ring. In some embodiments, -Cy-is optionally substituted phenylene. In some embodiments, -Cy-is 2, 6-dimethoxy-1, 4-phenylene. In some embodiments, -Cy-is 2,4, 6-trimethoxy-1, 3-phenylene. In some embodiments, R PS Is thatIn some embodiments, R PS is-SH.
In some embodiments, R a2 Is thatIn some embodiments, R a2 Is->In some embodiments, R a2 Is->In some embodiments, R a2 Is->In some embodiments, -C (R a2 )(R a3 ) -is->
In some embodiments, provided compounds, e.g., amino acids, are selected from the group consisting of:
in some embodiments, R a2 R in the above-mentioned compound a2 (non-Hydrogen radical attached to the alpha carbon)A bolus).
In some embodiments, the present disclosure provides a compound having the structure:
wherein:
ring a is an optionally substituted 3 to 10 membered ring;
n is 0 to 6;
m is 0 to 6; and is also provided with
Each other variable is independently as described herein.
In some embodiments, m is 0. In some embodiments, m is 1 to 6.
In some embodiments, the present disclosure provides a compound having the structure:
wherein:
ring a is an optionally substituted 3 to 10 membered ring;
n is 0 to 6;
m is 0 to 6; and is also provided with
Each other variable is as described herein.
In some embodiments, m is 0. In some embodiments, m is 1 to 6.
In some embodiments, the present disclosure provides a compound having the structure:
wherein:
ring a is an optionally substituted 3 to 10 membered ring;
n is 0 to 6; and is also provided with
Each other variable is as described herein.
In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 6. In some embodiments, n is 0, 1, or 2.
In some embodiments, m is 0. In some embodiments, m is 1. In some embodiments, m is 2. In some embodiments, m is 3. In some embodiments, m is 4. In some embodiments, m is 5. In some embodiments, m is 6. In some embodiments, m is 1, 2, or 3.
In some embodiments, ring a is a ring as described herein. In some embodiments, ring a is 3-membered. In some embodiments, ring a is 4 membered. In some embodiments, ring a is 5 membered. In some embodiments, ring a is 6 membered. In some embodiments, ring a is 7 membered. In some embodiments, ring a is 8 membered. In some embodiments, ring a is 9 membered. In some embodiments, ring a is 10 membered. In some embodiments, ring a is saturated. In some embodiments, ring a is partially unsaturated. In some embodiments, ring a is aromatic. In some embodiments, ring a has no additional heteroatoms other than nitrogen atoms. In some embodiments, the ring is unsubstituted. In some embodiments, ring a is substituted with one or more halogens. In some embodiments, ring a is substituted with one or more-F. In some embodiments, ring a has carbon substituted with two-F. In some embodiments, -C (O) R PS At the 2' bit (N at the 1 bit). In some embodiments, -C (O) R PS At the 3' position. In some embodiments, -C (O) R PS At the 4' position. In some embodiments, -C (O) R PS Attached to a chiral centre, for example a chiral carbon atom. In some embodiments, the chiral center is R. In some embodiments, the chiral center is S. In some embodiments, ring a is attached to- (CH) at the chiral carbon (which is R) 2 ) n-bonding. In some embodimentsRing a is bound to- (CH) at the chiral carbon (which is S) 2 ) n-bonding. In some embodiments, - (CH) 2 ) N-at 2 bits (N at 1 bit). In some embodiments, - (CH) 2 ) N-at 3 bits (N at 1 bit). In some embodiments, - (CH) 2 ) N-at 4 bits (N at 1 bit).
In some embodiments, ring a is substituted. In some embodiments, the substituents on ring a have suitable properties for a variety of uses, such as volume. In some embodiments, the substituents are independently selected from halogen, -R, -CF 3 、-N(R) 2 -CN and-OR, wherein each R is independently C optionally substituted with one OR more-F 1-6 Aliphatic series. In some embodiments, the substituents are independently selected from halogen, C 1-5 Linear, branched OR cyclic alkyl, -OR (wherein R is C 1-4 Linear, branched or cyclic alkyl), fluorinated alkyl, -N (R) 2 (wherein each R is independently C 1-6 Linear, branched, or cyclic alkyl), or-CN. In some embodiments, the substituents are selected from halogen, C having 0 to 4 heteroatoms 5-6 An aromatic ring, an optionally substituted 3-to 10-membered cycloalkyl or a heteroalkyl ring having 1 to 4 heteroatoms. In some embodiments, the substituent is halogen. In some embodiments, it is-F. In some embodiments, it is-Cl. In some embodiments, it is-Br. In some embodiments, it is-I. In some embodiments, the substituent is optionally substituted C 1-4 An alkyl group. In some embodiments, the substituent is C 1-4 An alkyl group. In some embodiments, it is methyl. In some embodiments, it is ethyl. In some embodiments, it is i-Pr. In some embodiments, the substituent is C 1-4 A haloalkyl group. In some embodiments, the substituent is C optionally substituted with one or more-F 1-4 An alkyl group. In some embodiments, it is-CF 3 . In some embodiments, it is-CN. In some embodiments, it is-OR, wherein R is optionally substituted C 1-4 An alkyl group. In some embodiments, it is-OR, where R is C 1-4 An alkyl group. In some embodiments, it is-OR, where R is C 1-4 A haloalkyl group. In some embodiments, it is-OR, wherein R is C optionally substituted with one OR more-F 1-4 An alkyl group. In some embodiments, it is-OCF 3 。
In some embodiments, ring a is or comprises an optionally substituted saturated monocyclic ring. In some embodiments, ring a is or comprises an optionally substituted partially unsaturated monocyclic ring. In some embodiments, ring a is or comprises an optionally substituted aromatic monocyclic ring. In some embodiments, ring a is optionally substituted phenyl. In some embodiments, ring a is an optionally substituted 5-to 6-membered heteroaryl having 1-3 heteroatoms. In some embodiments, ring a is an optionally substituted 5-to 6-membered heteroaryl having 1 to 3 heteroatoms, wherein at least one heteroatom is nitrogen. In some embodiments, ring a is an optionally substituted 8-to 10-membered bicyclic ring having 1 to 6 heteroatoms. In some embodiments, ring a is an optionally substituted 8-to 10-membered bicyclic aromatic ring having 1 to 6 heteroatoms, wherein each monocyclic unit is independently a 5-to 6-membered aromatic ring optionally having 0 to 3 heteroatoms. In some embodiments, ring A is attached to- (CH) at a carbon atom 2 ) n-bonding. In some embodiments, ring A is attached to- (CH) at the nitrogen atom 2 ) n-bonding. In some embodiments, L aa wherein-Cy-or Ring A is optionally substituted and each substituent is independently selected from halogen, -R, -CF 3 、-N(R) 2 -CN and-OR, wherein each R is independently C optionally substituted with one OR more-F 1-6 Aliphatic series. In some embodiments, L aa In (2) is optionally substituted and each substituent is independently selected from halogen, C 1-5 Linear, branched OR cyclic alkyl, -OR (wherein R is C 1-4 Linear, branched or cyclic alkyl), fluorinated alkyl, -N (R) 2 (wherein each R is independently C 1-6 Linear, branched, or cyclic alkyl), or-CN.
In some embodiments, ring a is optionally substituted phenyl. In some embodiments, the present disclosure provides a compound of formula (la)A compound or salt thereof, wherein ring a is optionally substituted phenyl and each variable is as described herein.
In some embodiments, the present disclosure provides a composition havingA compound of the structure, or a salt thereof, wherein each variable is independently as described herein. In some embodiments, the present disclosure provides a composition havingA compound of the structure, or a salt thereof, wherein each variable is independently as described herein.
In some embodiments, the compound is selected from:
in some embodiments, the present disclosure provides a compound of formula (la)A compound or salt thereof, wherein ring a is optionally substituted phenyl and each variable is as described herein. In some embodiments, the compound is selected from:
in some embodiments, ring a is optionally substituted with 1 to 4A 5 or 6 membered heteroaryl of a heteroatom. In some embodiments, provided compounds haveA structure wherein Z is a carbon or heteroatom, ring Het is an optionally substituted 5 or 6 membered heteroaryl having 1 to 4 heteroatoms, and each other variable is independently as described herein. In some embodiments, the provided compounds are selected from:
in some embodiments, ring a is an 8-to 10-membered bicyclic aryl or heteroaryl ring having 1 to 5 heteroatoms. In some embodiments, ring a is a 10 membered bicyclic aryl ring. In some embodiments, ring a is an 8-membered bicyclic heteroaryl ring having 1 to 5 heteroatoms. In some embodiments, ring a is a 9 membered bicyclic heteroaryl ring having 1 to 5 heteroatoms. In some embodiments, ring a is a 10 membered bicyclic heteroaryl ring having 1 to 5 heteroatoms. In some embodiments, ring a is an optionally substituted 5-or 6-membered heteroaryl having 1 to 4 heteroatoms. In some embodiments, provided compounds have A structure wherein each of rings r1 and r2 is independently an optionally substituted 5 or 6 membered aryl or heteroaryl ring having 1 to 4 heteroatoms, and each other variable is independently as described herein. In some embodiments, the provided compounds have +.>A structure wherein Z is a carbon or heteroatom, rings r1 and r2 are each independently an optionally substituted 5 or 6 membered aryl or heteroaryl ring having 1 to 4 heteroatoms, and each other variable is independently as described herein. In some embodiments, the provided compounds are selected from:
in some embodiments, the present disclosure provides a composition havingA compound of the structure or a salt thereof. In some embodiments, -C (O) R PS is-C (O) -OtBu. In some embodiments, the present disclosure provides a composition havingA compound of the structure, or a salt thereof, wherein each variable is independently as described herein.
In some embodiments, the provided compounds are selected from:
in some embodiments, the present disclosure provides a composition havingA compound of the structure, or a salt thereof, wherein each variable is independently as described herein. In some embodiments, the present disclosure provides a composition having ∈>A compound of the structure, or a salt thereof, wherein each variable is independently as described herein.
In some embodiments, the provided compounds are selected from:
in some embodiments, the provided compounds are amino acids. In some embodiments, the provided compounds are protected amino acids. In some embodiments, the provided compounds are protected and/or activated amino acids. In some embodiments, the provided compounds are suitable.
In some embodiments, the ring moiety, e.g., -Cy-, R (including those formed by R groups taken together), etc., is monocyclic. In some embodiments, the ring moiety is bicyclic or polycyclic. In some embodiments, the monocyclic ring is an optionally substituted 3 to 10 (3, 4, 5, 6, 7, 8, 9 or 10, 3 to 8, 3 to 7, 4 to 6, 5 to 6, etc.) membered saturated, partially unsaturated or aromatic ring having 0 to 5 heteroatoms. In some embodiments, each monocyclic ring unit of a bicyclic or polycyclic ring moiety is independently an optionally substituted 3 to 10 (3, 4, 5, 6, 7, 8, 9 or 10, 3 to 8, 3 to 7, 4 to 6, 5 to 6, etc.) membered saturated, partially unsaturated or aromatic ring having 0 to 5 heteroatoms.
In some embodiments, each heteroatom is independently selected from oxygen, nitrogen, sulfur, phosphorus, and silicon. In some embodiments, each heteroatom is independently selected from oxygen, nitrogen, and sulfur.
In some embodiments, L a1 Is a covalent bond. In some embodiments, the compound of formula PA has the structure NH (R a1 )-C(R a2 )(R a3 )-L a2 -COOH。
In some embodiments, L a2 Is a covalent bond. In some embodiments, the compound of formula PA has the structure NH (R a1 )-C(R a2 )(R a3 )-L a2 -COOH。
In some embodiments, L a1 Is a covalent bond, and L a2 Is a covalent bond. In some embodiments, the compound of formula PA has the structure NH (R a1 )-C(R a2 )(R a3 )-COOH。
In some embodiments, the amino acid is suitable for stapling. In some embodiments, the amino acid comprises a terminal olefin.
In some embodiments, the amino acid has NH (R a1 )-L a1 -C(-L aa -COOH)(R a3 )-L a2 -COOH structure or a salt thereof, wherein each variable is independently as described in the present disclosure. In some embodiments, L aa is-L am1 -N(R’)-L am2 -, wherein each variable is as described herein. In some embodiments, L am1 And L am2 Each is an optionally substituted divalent C 1-6 Aliphatic series. In some embodiments, L am1 And L am2 Each is a divalent C 1-6 Aliphatic series. In some embodiments, L am1 And L am2 Each is an optionally substituted divalent C 1-6 An alkyl group. In some embodiments, L am1 And L am2 Each is a divalent C 1-6 An alkyl group. In some embodiments, L am1 And L am2 Each is an optionally substituted divalent straight chain C 1-6 An alkyl group. In some embodiments, L am1 And L am2 Each is a divalent straight chain C 1-6 An alkyl group. In some embodiments, L am1 is-CH 2 -. In some embodiments, L am2 Is a covalent bond. In some embodiments, L am2 is-CH 2 -. In some embodiments, L am1 And L am2 Both are-CH 2 -. In some embodiments, L am1 is-CH 2 -, and L am2 Is a covalent bond. In some embodiments, -N (R'), -is-N (Et) -. In some embodiments, -N (R'), -is-N (CH) 2 CF 3 ) -. In some embodiments, L aa is-L am1 -Cy-L am2 -, wherein each variable is as described herein. In some embodiments, -Cy-is optionally substituted phenyl. In some embodiments, -Cy-is an optionally substituted 5-to 6-membered heteroaryl having 1 to 4 heteroatoms.
In some embodiments, the compound isOr a salt thereof. In some embodiments, the compound is +.>Or a salt thereof. In some embodiments, the compound isOr a salt thereof. In some embodiments, the compound is +.>Or a salt thereof. In some embodiments, the compound is +.>Or a salt thereof. In some embodiments, the compound isOr a salt thereof. In some embodiments, the compound is +.>Or a salt thereof. In some embodiments, the compound is +.>Or a salt thereof. In some embodiments, the compound is +. >Or a salt thereof. In some embodiments, the compound is +.>Or a salt thereof. Such compounds are particularly useful as amino acid residues in peptides, including stapled peptides.
In some embodiments, the present disclosure provides compounds, e.g., peptides, comprising residues of a compound of formula PA or a salt form thereof. In some embodiments, the residue has the formula-N (R a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C (O) -structure or salt form thereof, wherein each variable is independently as described herein. In some embodiments, the residue has the formula-N (R a1 )-L a1 -C(-L aa -COOH)(R a3 )-L a2 -C (O) -structure or salt form thereof, each of whichThe individual variables are independently as described herein. For example, in some embodiments, the residue isOr a salt form thereof. In some embodiments, the residue is +.>Or a salt form thereof. In some embodiments, the residue is +.>Or a salt form thereof. In some embodiments, the residue is +.>Or a salt form thereof. In some embodiments, the residue is +.>Or a salt form thereof. In some embodiments, the residue is +.>Or a salt form thereof. In some embodiments, the residue is +.>Or a salt form thereof. In some embodiments, the residue is +.>Or a salt form thereof. In some embodiments, the residue is +.>Or a salt form thereof.
Certain amino acids and moieties are described in WO 2022/020651 and WO 2022/020652, wherein the amino acids and moieties of each are independently incorporated herein by reference and can be used in accordance with the present disclosure.
In some implementationsIn embodiments, the amino acid or structural portion of an amino acid or agent (e.g., peptide) is selected from the following. N end cap (N-Term) through R 1 Amino (R) to the first amino acid (AA 1) 1 ) And (5) connection. In some embodiments, the N-Term cap may be considered as part of AA1 as appropriate. From here on, each carboxylate (R 2 ) Amino group (R) of subsequent amino acid 1 ) Linking until the carboxylate of the last amino acid (R 2 ) R with C-terminal group 1 And (5) connection. For any device having a branch point (R 3 ) And the amino acids of the branching monomers are shown in brackets, R of the monomers in brackets 1 R to amino acids 3 And (5) connection. For a device having two potential branch points (R 3 And R is 4 ) If two branches are shown, R of the first branch 1 And R is R 3 Is connected, and R of the second branch 1 And R is R 4 And (5) connection. For any pair of amino acids terminating at 3, R for each of these amino acids 3 The groups are all linked to each other. Likewise, for any pair of amino acids ending in the 3 name, R of these amino acids 3 The groups are linked to each other. For any inclusion having R 3 Branched amino acid pairs of groups and one of which comprises a group comprising R 1 And R is 2 Sequences of branching monomers of both radicals, R 1 To branched amino acids adjacent thereto in the sequence, and R of the branched monomer 2 R of groups with amino acids not provided for by branching monomers 3 And (5) connection. For example, in peptides having one of Cys, hCys, pen or aMeC at position 10 and one of Cys, hCys, pen or aMeC at position 14 as well, and a branching group other than amino acid residue 10, R of the branching group 1 R to an amino acid residue at position 10 3 R of the branching groups being taken together 2 R to an amino acid residue at position 14 3 Tied together. For any having a containing R 3 And no other amino acids linked thereto by the above-described manner, R 3 =h. Typically, all residues with terminal olefins are linked by ring closure metathesis (stapling). Some examples are provided in tables E2 and E3. In some embodiments, the present disclosure provides an agent, e.g., a peptide, e.g., a stapled peptide, comprising one or more amino acid residues selected from the group consisting of.
Tables a-iv certain useful compounds or moieties.
Tables A-IV (follow; some parts may be present in [ ])
Some moieties may be used as, for example, lys analogs, branch point amino acid residues or non-RCM stapled amino acid residues
Some portions may be used, for example, as stapled amino acid residues (e.g., RCM for other stapling techniques)
Some moieties may be used as, for example, aromatic amino acid residues
Certain portions may be used as amino acid residues
Some moieties may be used, for example, as amino acid residues (e.g., D-amino acid residues, homologous amino acid residues, alkyl (e.g., methyl) amino acid residues, etc.)
Some portions may be used as, for example, amino acid residues (e.g., alkyl amino acid residues, hydrophobic amino acid residues, etc.), for example
Some portions may be used as, for example, amino acid residues (e.g., polar amino acid residues, basic amino acid residues, etc.)
Some portions may be used as, for example, amino acid residues (e.g., acidic amino acid residues, non-aromatic amino acid residues, etc.)
Certain parts (e.g. parts used in [ ] in various medicaments)
Certain moieties (e.g., moieties used in [ ] in various agents, amino acid residues, etc.)
In some embodiments, there are two parts in brackets, e.g., [ Ac-dPEG2]R of the former is usually 1 R with the latter 1 And (5) connection. For example, in [ Ac-dPEG2]R of Ac 1 R with dPEG2 1 And (5) connection. R of dPEG2 2 Can be linked to other moieties, e.g. in [ Ac-dPEG2 ]]In Lys, R of dPEG2 2 R of Lys 3 And (5) connection.
In some embodiments, the present disclosure provides an agent, e.g., a peptide agent (in various embodiments, a stapled peptide agent), comprising a moiety selected from the table above. In some embodiments, the residue is stapled, e.g., forms a staple-like structure with another moiety. In some embodiments, the medicament comprises a staple-like structure formed between two portions, each independently selected from the list. In some embodiments, the staple-like structure comprises a double bond. In some embodiments, the staple-like structure comprises an E double bond. In some embodiments, the staple-like structure comprises a Z double bond. In some embodiments, the double bond is converted to another moiety, such as by hydrogenation to a saturated bond, by epoxidation to an epoxide, and the like. In some embodiments, a moiety (e.g., an amino acid residue) comprises two groups that can be used for stapling. In some embodiments, the amino acid residue comprises two groups for stapling, such as B3, B4, B5, B6, dap7Gly, dap7Pent, dapAc7EDA, dapAc7PDA, dap7Abu, and the like. In some embodiments, the N-terminal group (e.g., 4-pentenyl, 5-hexenyl, etc.) can be considered part of the first amino acid residue for stapling. In some embodiments, amino acid residues having an N-terminal group (e.g., 4-pentenyl, 5-hexenyl, etc.), such as 4-pentenyl-PL 3, 5-hexenyl-PL 3, etc., comprise two groups, e.g., two double bonds, for stapling. In some embodiments, the group for stapling is a double bond. In some embodiments, each group for stapling is independently a double bond. In some embodiments, for stapling The radical being a double bond and the other not a double bond (e.g. amino or R 3 Or comprises R 3 Is a group of (2). In some embodiments, the agent comprises two or more residues, each independently comprising two or more groups (e.g., double bonds), for stapling (e.g., 5 hexenyl-PL 3-Asp-alylgy-B5-Asp-3 COOHF-Ala-Phe-Leu-PyrS 2-2F3 MeF-BztA-gin-NH 2 or a salt thereof (ESP-1), 4 pentenyl-PL 3-Asp-alylgy-B5-Asp-3 COOHF-Ala-Phe-Leu-PyrS 2-2F3 MeF-BztA-gin-NH 2 or a salt thereof (ESP-2), and the like). In some embodiments, the agent comprises two or more amino acid residues, each amino acid residue independently bonded to two staple-like structures (e.g., 5 hexenyl-PL 3-Asp-all gly-B5-Asp-3 COOHF-Ala-Phe-Leu-PyrS 2-2F3 MeF-BztA-gin-NH 2 (ESP-1) or a salt thereof, 4 pentenyl-PL 3-Asp-all gly-B5-Asp-3 COOHF-Ala-Phe-Leu-PyrS 2-2F3 MeF-BztA-gin-NH 2 or a salt thereof (ESP 2), etc., wherein double bonds are used to form a staple-like structure; in some embodiments, the staple-like structure is formed by olefin metathesis; in some embodiments double bonds in the staple-like structure are further converted to saturated bonds (e.g., by hydrogenation)). In some embodiments, the agent (e.g., ESP-1, ESP-2, etc.) comprises two or more staple-like structures within a short sequence and provides a high stapling density, e.g., (i, i+2) and (i, i+3) staple-like structures bonded to the same amino acid residue. In some embodiments, the staple-like structures in the provided medicaments are more evenly distributed such that for any amino acid residue bonded to two or more staple-like structures, one and only one is (i, i+2) or (i, i+3). Thus, in some embodiments, the agent is not ESP-1 or ESP-2 (where ESP-1 and ESP-2 are not stapled, are stapled, or are post-stapled modified (e.g., hydrogenated to convert double bonds in a staple-like structure to single bonds)). In some embodiments, the agent comprises one and no more than one residue comprising two or more residues for stapling. In some embodiments, the agent comprises one and no more than one amino acid residue bonded to two staple-like structures. In some embodiments In one embodiment, the medicament comprises staple-like structures having different types of structures and/or staple-like structures formed by different types of transformations. For example, in some embodiments, the pharmaceutical agent comprises a staple-like structure that forms a structure that does not include olefin metathesis conversion and/or modification of carbon-carbon double bonds (e.g., hydrogenation). In some embodiments, such agents may provide improved properties, activity, design flexibility, manufacturing efficiency, and the like.
In some embodiments, the compound has a structure selected from the list above, wherein R 1 is-OH. In some embodiments, the compound has a structure selected from the list above, wherein R 1 is-H. In some embodiments, the compound is a compound having a structure selected from the above table, or a salt thereof, wherein R 1 is-H or an amino protecting group (e.g., fmoc, tBOC, etc.) and R 2 is-OH, a carboxyl protecting or activating group. In some embodiments, the compound is a compound having a structure selected from the above table, or a salt thereof, wherein R 1 is-H or an amino protecting group and R 2 is-OH. In some embodiments, the compound is a compound having a structure selected from the above table, or a salt thereof, wherein R 1 is-H and R 2 is-OH. In some embodiments, the compound is a compound having a structure selected from the above table, or a salt thereof, wherein R 1 is-H, R 2 is-OH and R 3 is-H. In some embodiments, R 3 is-H or a protecting group. In some embodiments, R 3 is-H. In some embodiments, the compound has a structure selected from the list above, wherein R 1 Is an amino protecting group such as Fmoc, tBOC, etc. In some embodiments, the compound has a structure selected from the list above, wherein R 1 Is an amino protecting group such as Fmoc, tBOC, etc., and R 2 is-OH, or-COR 2 Is an optionally substituted, protected or activated carboxyl group. In some embodiments, R 2 is-OH. In some embodiments, the amino acid residue has a structure selected from the list above, wherein R 1 And R is 2 Each independently represents a linking site (e.g., for a structureResidues have the structure->). In some embodiments, the agent, peptide, or stapled peptide comprises such an amino acid residue.
In some embodiments, the peptide comprises one or more residues selected from the amino acids of the above table. In some embodiments, the peptide comprises one or more residues of TfeGA. In some embodiments, the peptide comprises one or more residues of 2 COOHF. In some embodiments, the peptide comprises one or more residues of 3 COOHF.
The present disclosure provides, inter alia, peptides, including stapled peptides, comprising residues of the amino acids described herein. In some embodiments, the present disclosure provides methods comprising utilizing optionally protected and/or activated amino acids, as described herein. In some embodiments, the present disclosure provides methods for preparing peptides comprising utilizing amino acids that are generally protected and/or activated, as described herein. For example, in some embodiments, multiple amino groups are Fmoc protected for peptide synthesis (particularly for formation of backbone peptide bonds). In some embodiments, a plurality of side chain carboxylic acid groups are protected by t-Bu (-C (O) -O-tBu).
In some embodiments, the present disclosure provides methods comprising replacing one or more acidic amino acid residues (e.g., asp, glu, etc.) in a first compound with a provided amino acid residue (e.g., tfeGA, 2COOHF, 3COOHF, etc.) each independently to provide a second compound. In some embodiments, each of the first and second compounds is independently a peptide or independently comprises a peptide. In some embodiments, the second compound provides improved properties and/or activity (e.g., lipophilicity, logD, etc.) as compared to the first compound. In some embodiments, the second compound provides one or more comparable or improved other properties and/or activities (e.g., solubility and/or target binding) in addition to the improved properties (e.g., lipophilicity) as compared to the first compound.
In some embodiments, the agent (e.g., peptide, stapled peptide, suture peptide, etc.) has a mass of less than about 5000 daltons. In some embodiments, the agent has a mass greater than or equal to about 900 daltons and less than about 5000 daltons. In some embodiments, the agent has a mass greater than or equal to about 1500 daltons and less than about 5000 daltons. In some embodiments, the agent has a mass greater than or equal to about 2000 daltons and less than about 5000 daltons. In some embodiments, the agent has a mass greater than or equal to about 2500 daltons and less than about 5000 daltons. In some embodiments, the agent has a mass greater than or equal to about 1000 daltons and less than about 3000 daltons. In some embodiments, the mass of the agent is greater than or equal to about 1500 daltons and less than about 3000 daltons. In some embodiments, the agent has a mass greater than or equal to about 1500 daltons and less than about 2500 daltons. In some embodiments, the agent has a mass greater than or equal to about 1600 daltons and less than about 2200 daltons. In some embodiments, the agent has a mass of no more than about 900 daltons. In some embodiments, the agent has a mass of no more than about 500 daltons. In some embodiments, the agent has a mass of no more than about 300 daltons. In some embodiments, the agent has a mass of no more than about 200 daltons.
Characterization of
In some embodiments, the agent (e.g., peptide) is characterized with respect to, for example, one or more characteristics, such as binding characteristics, e.g., with respect to a particular target of interest (e.g., β -catenin or a portion thereof), stability characteristics (e.g., in solution or in dry form), cell permeability characteristics, solubility, lipophilicity, and the like.
In some embodiments, the binding features may be or include the following: specificity, affinity, binding rate, dissociation rate, etc., optionally under (or within the scope of) specified conditions, such as, for example, concentration, temperature, pH, cell type, presence or level of a particular competitor, etc.
As will be appreciated by those of skill in the art, the evaluation of features described herein may involve comparison to an appropriate reference (e.g., positive or negative control), which may be a contemporaneous reference in some embodiments, or may be a historical reference in some embodiments.
In some embodiments, the desired feature may be, for example: binding to the desired target (e.g., dissociation constant (K D ) Is at least less than about 1. Mu.M, and preferably K D Less than about 50 nM); cell permeability (e.g., as measured by fluorescence-based assay or mass spectrometry of cell fractions, etc.); solubility (e.g., can dissolve less than about 1000uM of an agent, or can dissolve less than about 500uM of an agent, or can dissolve less than about 100uM of an agent, or less than about 50uM of an agent, or less than about 35uM of an agent); activity (e.g., modulating one or more functions of a target), which may be measured at a cellular reporter (e.g., where the IC50 is at a concentration of less than about 1 μM, less than about 500nM, less than about 50nM, less than about 10nM, etc.), an animal model (e.g., a variety of animal models for a condition, disorder, or disease, e.g., a mouse melanoma model Braf) V600E /Pten --/-- And Braf V600E /Pten --/-- CAT-STA) and/or in the subject; stability, which can be assessed using a number of assays (e.g., in a rat pharmacokinetic study (e.g., by oral, iv, ip, etc.) with a terminal half-life greater than a suitable time (e.g., 1 hour); low toxicity, which can be assessed by a number of assays (e.g., standard ADME/toxicity assays); and/or low levels of cytotoxicity (e.g., low levels of lactate dehydrogenase (lactate dehydrogenase, LDH) released from cells when treated with a suitable concentration (e.g., about 10 μm) of peptide). In some embodiments, the agents of the invention comprise an affinity of less than about 10nM, e.g., an IC50 of 7nM.
In some embodiments, provided agents can bind to a target (e.g., β -catenin) with an EC50 of no greater than about 2000nM. In some embodiments, the EC50 is not greater than about 1500nM. In some embodiments, the EC50 is not greater than about 1000nM. In some embodiments, the EC50 is not greater than about 500nM. In some embodiments, the EC50 is not greater than about 300nM. In some embodiments, the EC50 is not greater than about 200nM. In some embodiments, the EC50 is not greater than about 100nM. In some embodiments, the EC50 is not greater than about 75nM. In some embodiments, the EC50 is not greater than about 50nM. In some embodiments, the EC50 is not greater than about 25nM. In some embodiments, the EC50 is not greater than about 10nM. In some embodiments, the EC50 is not greater than about 5nM. In some embodiments, EC50 is measured by fluorescence polarization, as described in the examples.
In some embodiments, the present disclosure provides agents, e.g., stapled peptides, with suitable solubility for a variety of purposes. In some embodiments, the provided agent (e.g., in PBS) has a solubility of about or at least about 5 to 100uM (e.g., about or at least about 5, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, or 100 uM). In some embodiments, the solubility is about or at least about 25uM. In some embodiments, the solubility is about or at least about 30uM. In some embodiments, the solubility is about or at least about 40uM. In some embodiments, the solubility is about or at least about 50uM. In some embodiments, the provided agent (e.g., a stapled peptide) is a protein that binds in serum; in some embodiments, at least about 85%, 90%, or 95% of its proteins are bound in serum. In some embodiments, more than 95% of the protein in the provided agent is bound in serum.
In some embodiments, the provided agent can cross the cell membrane of an animal cell. In some embodiments, the provided agent may cross the cell membrane of a human cell.
The provided agents may be particularly associated with motifs, residues or polypeptides. In some embodiments, the provided agent binds to β -catenin. In some embodiments, the dissociation constant (K D ) From about 1nM to about 1uM. In some embodiments, K D Not greater than about 1uM. In some embodiments, K D No greater than about 500nM. In some embodiments, K D No greater than about 250nM. In some embodiments, K D No greater than about 100nM. In some implementationsIn embodiments, K D No greater than about 50nM. In some embodiments, K D No greater than about 25nM. In some embodiments, K D No greater than about 10nM. In some embodiments, K D No greater than about 5nM. In some embodiments, K D No greater than about 1nM. As will be appreciated by those of skill in the art, a variety of techniques are available and may be used to measure K in accordance with the present disclosure D . In some embodiments, K D Measured by surface plasmon resonance (Surface Plasmon Resonance, SPR) as shown herein.
In some embodiments, the provided agent binds to a polypeptide having the sequence SEQ ID No. 2 or comprising SEQ ID No. 2 or a fragment thereof:
in some embodiments, the provided agent has one or more or all of the following interactions with β -catenin:
direct interaction (), water mediated [ ], non-polar contact { }, and
LQIL { AY } (G) { NQ } ES (K) LIILA (residues 301 to 317 of the Uniprot P35222 sequence) (SEQ ID NO: 3)
SRVL { (K) V } LS { V } CSSN (residues 341 to 353 of the Uniprot P35222 sequence) (SEQ ID NO: 4)
RLV { QN } C { L } (W) TL { R } (N) LSDA (residues 376 to 391 of the Uniprot P35222 sequence) (SEQ ID NO: 5)
LGSD [ D ] I (N) { V } V { TC } AAGI (residues 409 to 423 of Uniprot P35222 sequence) (SEQ ID NO: 6)
In some embodiments, the agent (e.g., peptide) binds to β -catenin and interacts with one or more residues that are or correspond to: at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or at least eight, or at least nine, or at least ten, or at least eleven, or at least twelve, or at least thirteen, or at least fourteen, or at least fifteen, or at least sixteen, or at least seventeen, or at least eighteen, or at least nineteen, or at least twenty of the following amino acid residues at the positions shown in SEQ ID No. 1: a305 Y306, G307, N308, Q309, K312, K345, V346, V349, Q379, N380, L382, W383, R386, N387, D413, N415, V416, T418, and C419. In some embodiments, the agent (e.g., peptide) binds to β -catenin and interacts with one or more residues that are or correspond to: SEQ ID NO:1, or at least three, or at least four, or at least five, or at least six, or seven of the following amino acid residues at the positions shown in 1: g307 K312, K345, W383, R386, N387, D413, and N415. In some embodiments, the agent (e.g., peptide) binds to β -catenin and interacts with one or more residues that are or correspond to: SEQ ID NO:1, or at least three, or at least four, or at least five, or at least six, or seven of the following amino acid residues at the positions shown in 1: g307 K312, K345, W383, N387, D413, and N415.
In some embodiments, the provided agent interacts with β -catenin at one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, or 9) of G307, K312, K345, Q379, L382, W383, N387, N415, and V416. In some embodiments, the provided agent interacts with β -catenin at one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of Y306, G307, K312, K345, Q379, L382, W383, N387, N415, and V416. In some embodiments, the provided agent interacts with β -catenin at one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of G307, K312, K345, Q379, L382, W383, R386, N387, N415, and V416. In some embodiments, the provided agent interacts with β -catenin at one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) of Y306, G307, K312, K345, Q379, L382, W383, R386, N387, N415, and V416. In some embodiments, the provided agent interacts with β -catenin at one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) of Y306, G307, K312, K345, V349, Q379, L382, W383, R386, N387, N415, and V416. In some embodiments, the provided agent interacts with β -catenin at one or more (e.g., 1, 2, 3, 4, 5, 6, or 7) of G307, K312, K345, W383, R386, N387, D413, and N415. In some embodiments, the provided agent interacts with β -catenin at one or more (e.g., 1, 2, 3, 4, 5, 6, or 7) of G307, K312, K345, W383, N387, D413, and N415. In some embodiments, the provided agents interact with β -catenin at one or both of K312 and R386. In some embodiments, the provided agent interacts with G307. In some embodiments, the provided agent interacts with K312. In some embodiments, the provided agents interact with β -catenin at one or more of K345, W383, D413, and N415. In some embodiments, the provided agents interact with β -catenin at one or more of K345 and W383. In some embodiments, the provided agents interact with β -catenin at one or more of D413 and N415. In some embodiments, the provided agent interacts with Y306. In some embodiments, the provided agent interacts with G307. In some embodiments, the provided agent interacts with K312. In some embodiments, the provided agent interacts with K345. In some embodiments, the provided agent interacts with V349. In some embodiments, the provided agent interacts with Q379. In some embodiments, the provided agent interacts with L382. In some embodiments, the provided agent interacts with W383. In some embodiments, the provided agent interacts with R386. In some embodiments, the provided agent interacts with N387. In some embodiments, the provided agent interacts with D413. In some embodiments, the provided agent interacts with N415. In some embodiments, the provided agent interacts with V416.
In some embodiments, the provided agents interact with one or more of the amino acid residues that are or correspond to: k312, R386, K345 and W383 of SEQ ID NO. 1. In some embodiments, the provided agents interact with one or more of the amino acid residues that are or correspond to: k312 and R386 of SEQ ID NO. 1. In some embodiments, interactions with amino acid residues can be assessed by mutation of such amino acid residues (e.g., mutation of K, R, etc. to D, E, etc.).
Those of skill in the art having read the present disclosure will appreciate that in some embodiments, interactions with β -catenin can be assessed by contacting an agent with full-length β -catenin or a portion of β -catenin. In some embodiments, a portion of the β -catenin comprises residues of the interactions described above. In some embodiments, a portion of the β -catenin is SEQ ID NO. 2 or comprises SEQ ID NO. 2. In some embodiments, a portion of the β -catenin is expressed with a tag (e.g., for purification, detection, etc.). In some embodiments, the tag is a fluorescent tag. In some embodiments, the tag is used for detection. In some embodiments, the tag is used for purification and detection. In some embodiments, the tag is a purification tag. In some embodiments, the tag is or comprises biotin. Many other types of labels are available in the art and may be used in accordance with the present disclosure.
In accordance with the present disclosure, a variety of techniques may be used to characterize and/or evaluate the provided techniques (e.g., agents (e.g., various peptides), compositions, methods, etc.)). As described herein, in some embodiments, a useful technique is or includes fluorescence polarization. In some embodiments, available techniques evaluate LogP or LogD. In some embodiments, a useful technique is or comprises a CHI log d assay. In some embodiments, the solubility can be assessed using techniques. In some embodiments, a useful technique is or comprises NanoBRET. In some embodiments, a useful technique is a reporter assay (e.g., DLD1 reporter assay) or comprises a reporter assay (e.g., DLD1 reporter assay). In some embodiments, the available technology is or contains alphascreen. Some of the available schemes are described in the embodiments. Those skilled in the art will appreciate that such schemes (e.g., depending on particular conditions, agents, purposes, etc.) may be appropriately modified.
Production of
Various techniques for producing the provided medicaments are known in the art. For example, various techniques for preparing small molecules, peptides (including stapled peptides) may be utilized in accordance with the present disclosure. Those skilled in the art who review this disclosure will well understand which such techniques are applicable in accordance with this disclosure in which aspects of this disclosure.
Stapling can be performed during and/or after peptide chain synthesis. In some embodiments, the present disclosure provides an unpinned peptide agent whose sequence is the sequence described in table E2 or table E3. In some embodiments, the amino acid residue is optionally protected for peptide synthesis (e.g., peptide synthesis using Fmoc protected amino acids, wherein certain side chains may be protected). In some embodiments, one or more stapling is achieved by olefin metathesis. In some embodiments, two or more staplers are formed by one olefin metathesis process. In some embodiments, the present disclosure provides a stapled peptide agent described in table E2 or table E3, or a salt thereof (e.g., a pharmaceutically acceptable salt thereof). In some embodiments, the present disclosure provides stereoisomers of the stapled peptide agents described in table E2 or table E3, or salts thereof (e.g., pharmaceutically acceptable salts thereof). In some embodiments, the present disclosure provides E/Z stereoisomers of the stapled peptide agents described in table E2 or table E3, or salts thereof (e.g., pharmaceutically acceptable salts thereof). In some embodiments, from the N to C direction, the olefinic double bond in a first staple-like structure comprising such a bond is Z, and the olefinic double bond in a second staple-like structure comprising such a bond is E (Z-E); in some embodiments, it is (Z-Z); in some embodiments, it is (E-Z); in some embodiments, it is (E-E). In some embodiments, from the N to C direction, the olefinic double bond in the first (i, i+2), (i, i+3), or (i, i+4) staple-like structure comprising such a bond is Z, and the olefinic double bond in the first (i, i+7) staple-like structure comprising such a bond is E (Z-E); in some embodiments, it is (Z-Z); in some embodiments, it is (E-Z); in some embodiments, it is (E-E). In some embodiments, the medicament comprises an olefinic double bond in the third staple-like structure, and it is E; in some embodiments, it is Z. In some embodiments, the medicament comprises an olefinic double bond in the fourth staple-like structure, and it is E; in some embodiments, it is Z.
In some embodiments, one or more or all of the staple-like structures are formed after chain extension. In some embodiments, one or more or all of the staple-like structures are formed during chain extension. In some embodiments, one or more or all of the staple-like structures are formed by metathesis after chain extension. In some embodiments, one or more or all of the staple-like structures are formed by metathesis during chain extension.
In some embodiments, the present disclosure provides methods comprising
a) Preparing a first compound comprising two moieties, each independently comprising an olefinic double bond;
b) Stapling the two parts to form a first formed staple-like structure by olefin metathesis between the olefin double bond in one part and the olefin double bond in the other part, thereby providing a second compound;
c) Adding one or more additional moieties to the second compound to provide a third compound comprising two moieties, each independently comprising an olefinic double bond; and
d) Two of the third compounds are stapled to form a second formed staple-like structure by olefin metathesis between an olefin double bond in one moiety and an olefin double bond in the other moiety, thereby providing a fourth compound.
In one placeIn some embodiments, the moiety is an amino acid residue. In some embodiments, each moiety is independently an amino acid residue. In some embodiments, each moiety is independently an amino acid residue comprising a terminal olefin as described herein. In some embodiments, there are two olefinic double bonds in one portion of the first compound, for example. For example, in some embodiments, such moiety is B5. In some embodiments, the two moieties of the first compound are independently X 4 And X 11 . In some embodiments, the first formed staple-like structure is an (i, i+7) staple-like structure. In some embodiments, the first compound comprises-X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 -. In some embodiments, the first compound comprises-X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 -. In some embodiments, the first compound comprises a staple-like structure. In some embodiments, the staple-like structure is an (i, i+4) staple-like structure. In some embodiments, the staple-like structure is at X 10 And X is 14 Between them. In some embodiments, the olefinic double bond in the third compound is present in the first compound (e.g., the non-stapled olefinic double bond of B5). In some embodiments, one and only one amino acid residue comprising an olefinic double bond is added to the second compound. In some embodiments, the third compound is or comprises the following: -X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 -. In some embodiments, the third compound is or comprises the following: -X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 -. In some embodiments, the first and second formed staple-like structures are bonded to the same amino acid residue. In some embodiments, the first and second formed staplesThe spike-like structures are bonded to the same atom. In some embodiments, the second formed staple-like structure is an (i, i+2), (i, i+3), or (i, i+4) staple-like structure. In some embodiments, two moieties in the third compound are independently X 1 And X 4 . In some embodiments, a first formed staple-like structure (e.g., about 1.1:1, 1.2:1, 1.3:1, 1.4:1, 1.5:1, 2:1, 3:1, 4:1, 5:1, 10:1, 20:1, 30:1, 40:1, 50:1, or higher) is formed having E selectivity as described herein. In some embodiments, a second formed staple-like structure (e.g., about 1.1:1, 1.2:1, 1.3:1, 1.4:1, 1.5:1, 2:1, 3:1, 4:1, 5:1, 10:1, 20:1, 30:1, 40:1, 50:1, or higher) is formed having Z selectivity as described herein. In some embodiments, the synthesis may be performed on a solid support (e.g., solid phase peptide synthesis), and the compound or agent may be on a solid support. In some embodiments, stapling during chain extension or separately for one or more staple-like structures may provide advantages, such as increased selectivity, yield, purity, etc.
In some embodiments, two or more staple-like structures are formed in the metathesis reaction. In some embodiments, all of the staple-like structures formed by metathesis are formed in the metathesis reaction. In some embodiments, each such staple-like structure is formed by olefin metathesis of a terminal olefin. In some embodiments, the plurality of staple-like structures are formed after the full length of the peptide has been achieved. In some embodiments, one or more staple-like structures comprising double bonds are formed after the full length of the peptide has been achieved. In some embodiments, all staple-like structures comprising double bonds are formed after the full length of the peptide has been achieved. In some embodiments, the one or more staple-like structures formed by metathesis are formed after the full length of the peptide has been achieved. In some embodiments, all of the staple-like structures formed by metathesis are formed after the full length of the peptide has been achieved.
In some embodiments, step-wise stapling is performed wherein two or more staple-like structures are formed in two or more steps. In some embodiments, progressive stapling provides an improved level of selectivity to form a desired product (e.g., I-66) as compared to other compounds such as stereoisomers (e.g., I-66, I-67). In some embodiments, the improvement is about or at least about 1.5, 2, 2.5, 3, 4, 5, 6, 7, 8, 9, or 10 times. In some embodiments, improvement is assessed by comparing the percentage of the desired product in all relevant stereoisomers. In some embodiments, improvement is assessed by the ratio of the desired product to stereoisomers (e.g., I-66 to I-67). In some embodiments, the two staple-like structures comprising olefinic double bonds are formed in two separate steps. In some embodiments, the two staple-like structures formed by metathesis are formed in two separate steps. In some embodiments, two staple-like structures bonded to the same amino acid residue are formed in two separate steps. In some embodiments, two staple-like structures bonded to the same atom are formed in two separate steps. In some embodiments, two staple-like structures bonded to the same carbon atom are formed in two separate steps. In some embodiments, the two staple-like structures formed by B5 are formed in two separate steps. In some embodiments, the provided techniques include a third step of forming a third staple-like structure. In some embodiments, each staple-like structure is formed in a separate step. In some embodiments, the present disclosure provides a method for preparing a stapled peptide comprising:
1) Reacting the first reactive group with the second reactive group to form a first staple-like structure, wherein the first and second reactive groups are in two different amino acid residues; and
2) Reacting the third reactive group with a fourth reactive group to form a second staple-like structure, wherein the third and fourth reactive groups are in two different amino acid residues.
Alternatively or additionally, in some embodiments, the method includes reacting a fifth reactive group with a sixth reactive group to form a third staple-like structure, wherein the fifth and sixth reactive groups are in two different amino acid residues. In some embodiments, the third staple-like structure is formed before the first and second staple-like structures.
In some embodiments, the first staple-like structure is formed by a metathesis reaction. In some embodiments, the first and second reactive groups are each independently a double bond or comprise a double bond. In some embodiments, the first and second reactive groups are each independently a terminal olefin. In some embodiments, the first staple-like structure is formed by olefin metathesis. In some embodiments, the first staple-like structure is an (i, i+7) staple-like structure. In accordance with the present disclosure, a variety of metathesis techniques may be utilized to form the first staple-like structure. In some embodiments, the metathesis reaction is carried out in the presence of a catalyst. In some embodiments, the catalyst is a Hoveyda-Grubbs M720 catalyst (CAS 301224-40-8). In some embodiments, the first staple-like structure is at X 4 And X is 11 Between them.
In some embodiments, the second staple-like structure is formed by a metathesis reaction. In some embodiments, the third and fourth reactive groups are each independently a double bond or comprise a double bond. In some embodiments, the third and fourth reactive groups are each independently a terminal olefin. In some embodiments, the second staple-like structure is formed by olefin metathesis. In some embodiments, the second staple-like structure is an (i, i+3) staple-like structure. In accordance with the present disclosure, a variety of metathesis techniques may be utilized to form the second staple-like structure. In some embodiments, the metathesis reaction is carried out in the presence of a catalyst. In some embodiments, the catalyst is Grubbs M102 catalyst (CAS 172222-30-9). In some embodiments, the second staple-like structure is at X 1 And X is 4 Between them.
In some embodiments, one of the first and second reactive groups, and one of the third and fourth reactive groups are in the same amino acid residue. In some embodiments, they are independently in side chains, and both side chains are bonded to the same atom. In some embodiments, both side chains are bonded to the same carbon atom (e.g., as in B5). In some embodiments, the first and second staple-like structures are bonded to the same amino acid residue. In some embodiments, they are bonded to the same atom. In some embodiments, they are bonded to the same carbon (e.g., in B5).
In some embodiments, the third staple-like structure comprises an amide group, such as-C (O) N (R ') -, wherein R' is as described herein. In some embodiments, the third staple-like structure comprises-C (O) NH-. In some embodiments, the third staple-like structure is an (i, i+4) staple-like structure. In some embodiments, one of the fifth and sixth reactive groups is or comprises an amino group or an activated form thereof and the other is or comprises an acidic group such as a carboxyl group or an activated form thereof. In some embodiments, the third staple-like structure is formed by an amidation reaction. In some embodiments, the third staple-like structure is not formed by a metathesis reaction. In some embodiments, the third staple structure does not comprise an olefinic double bond. A variety of amidation techniques are available and may be used herein. Other types of staple-like structures may also be used and constructed as described herein. See, e.g., the preparation of I-66, I-335, etc. in the examples. In some embodiments, the third staple-like structure is at X 10 And X is 14 Between them.
In some embodiments, one or more stapling steps are performed independently prior to achieving full length, as described herein. For example, in some embodiments, a third staple-like structure is formed before both of the two amino acid residues comprising the first and second reactive groups are installed. Alternatively or additionally, in some embodiments, the first staple-like structure is formed before both of the two amino acid residues comprising the third and fourth reactive groups are installed. In some embodiments, the composition comprises a firstThe amino acid residue of one of the first and second reactive groups is attached, but before the amino acid residue comprising the other of the first and second reactive groups is attached, to form a third staple-like structure. In some embodiments, the first staple-like structure is formed after the amino acid residue comprising one of the third and fourth reactive groups is installed, but before the amino acid residue comprising the other of the third and fourth reactive groups is installed. In some embodiments, two or more stapling steps are performed based on the location of the relevant staple-like structure and the direction of peptide synthesis, and one or more staple-like structures closer to the starting end are formed before one or more staple-like structures further from the starting end. In some embodiments, peptide synthesis is from the C-terminus to the N-terminus. In some embodiments, for the first and second staple-like structures, the staple-like structure with the two relevant residues installed first is formed first. For example, in C-terminal to N-terminal peptide synthesis, X 4 And X is 11 The staple-like structure between is X 1 And X is 4 The staple-like structure therebetween is formed before.
A variety of metal complexes or catalysts may be used for metathesis. For example, in some embodiments, the metal complex is a Grubbs catalyst. In some embodiments, it is in some embodiments a Hoveyda-Grubbs catalyst. In some embodiments, it is Grubbs im 102.Hoveyda-Grubbs M720 catalyst. In some embodiments, the catalyst provides product E/Z selectivity. As will be appreciated by those skilled in the art, the catalyst may be used at a variety of suitable levels, for example, about 1%, 2%, 3%, 4%, 5%, 10%, 20%, 25%, 30%, 40%, 50% mol or more.
In some embodiments, the present disclosure provides techniques for controlling the ratio of E/Z isomers of one or more or each olefinic double bond formed during olefin metathesis. In some embodiments, one or more or each olefinic double bond is formed at an isomer ratio of about 1.1:1, 1.2:1, 1.3:1, 1.4:1, 1.5:1, 2:1, 3:1, 4:1, 5:1, 10:1, 20:1, 30:1, 40:1, 50:1 or higher. In some embodiments, one or more or each olefinic double bond has an isomer ratio of about 1.1:1, 1.2:1, 1.3:1, 1.4:1, 1.5:1, 2:1, 3:1, 4:1, 5:1, 10:1, 20:1, 30:1, 40:1, 50:1 or higher in the product composition. In some embodiments, independently, it is about 1.5:1 or higher. In some embodiments, independently, it is about 2:1 or higher. In some embodiments, independently, it is about 3:1 or higher. In some embodiments, independently, it is about 4:1 or higher. In some embodiments, independently, it is about 5:1 or higher. In some embodiments, independently, it is about 6:1 or higher. In some embodiments, independently, it is about 7:1 or higher. In some embodiments, independently, it is about 8:1 or higher. In some embodiments, it is independently about 9:1 or higher. In some embodiments, independently, it is about 10:1 or higher. In some embodiments, it is independently about 20:1 or higher. In some embodiments, independently, it is about 30:1 or higher. In some embodiments, it is independently about 40:1 or higher. In some embodiments, independently, it is about 50:1 or higher. In some embodiments, the ratio is E:Z. In some embodiments, the ratio is Z: E.
In some embodiments, stapling produces one or more chiral centers. For example, in some embodiments, a chiral center may be formed when B5 forms two staple-like structures with two other amino acid residues. In some embodiments, the chiral center formed in the agent is R. In some embodiments, the chiral center formed in the agent is S. In some embodiments, the composition comprises two agents that are R and S at the chiral center. In some embodiments, chiral centers are formed that are stereoselective (e.g., diastereoselective when other chiral elements are present in the same molecule in some embodiments). In some embodiments, the selectivity is about or at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% (when the selectivity is 98%, 98% of all product molecules share the same stereochemistry at the chiral center). In some embodiments, in a composition described herein (e.g., a pharmaceutical composition), about or at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% of all molecules and salts thereof having the same composition (structure) share the same stereochemistry at the chiral center, e.g., at the chiral center bonded to two staple-like structures, e.g., in B5. In some embodiments, it is about or at least about 70%. In some embodiments, it is about or at least about 75%. In some embodiments, it is about or at least about 80%. In some embodiments, it is about or at least about 85%. In some embodiments, it is about or at least about 90%. In some embodiments, it is about or at least about 95%. In some embodiments, it is about or at least about 98%. In some embodiments, it is about or at least about 99%.
In some embodiments, the olefinic double bonds in the staple structure may be further modified. In some embodiments, the olefinic double bonds in the staple-like structure are hydrogenated, thus converting them to single bonds. In some embodiments, the modification is epoxidation. In some embodiments, the modification is halogenation. Those skilled in the art will appreciate that a variety of other modifications are suitable for olefinic double bonds and may be used in accordance with the present disclosure.
In some embodiments, the crude product composition is purified, for example, by chromatographic techniques such as liquid chromatography. In some embodiments, one or more product compositions are collected based on separate fractions, such as HPLC peaks, having the correct observed quality. In some embodiments, each product composition independently corresponds to a different peak having the correct observed quality (e.g., in some embodiments, by UV detection at a suitable wavelength, such as 220 nm). In some embodiments, the peak area of one or more or each product composition is independently about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more of the total peak area of all peaks having the correct mass. In some embodiments, it is about 5% or more. In some embodiments, it is about 10% or more. In some embodiments, it is about 20% or more. In some embodiments, it is about 25% or more. In some embodiments, it is about 30% or more. In some embodiments, it is about 40% or more. In some embodiments, it is about 50% or more. In some embodiments, the product composition comprises one isomer. In some embodiments, the product composition comprises two or more isomers (e.g., those that are insufficiently separable). In some embodiments, each product composition independently has a purity and/or a stereochemistry as described herein, e.g., in some embodiments, for one or more (e.g., 1,2, 3, 4, 5, or more) or each olefinic double bond in a staple-like structure, the ratio of the two stereoisomers is independently about 1.1:1, 1.2: 1,1.3: 1,1.4:1, 1.5:1, 2:1, 3:1, 4:1, 5:1, 10:1, 20:1, 30:1, 40:1, 50:1, 60:1, 70:1, 80:1, 90:1, 100:1, or more. In some embodiments, the ratio may be assessed by NMR, HPLC, or the like.
In some embodiments, certain stapled peptides and in particular cysteine stapled peptides may be provided in and/or produced by a biological system and have R with a provided reagent, for example, as described herein x -L s2 -R x Reagent reaction of a structure or salt thereof, wherein R x Can be reacted with-SH groups under suitable conditions. In some embodiments, each R x Is a suitable leaving group. In some embodiments, each R x Independently, -Br.
In some embodiments, the peptides are typically prepared on a solid phase on a synthesizer using Fmoc chemistry. In some embodiments, the present disclosure provides protected and/or activated amino acids for synthesis.
In some embodiments, the staple-like structure is formed by olefin metathesis. In some embodiments, the double bonds of the metathesis products are reduced/hydrogenated. In some embodiments, the CO 2 Extruded from the urethane portion of the staple-like structure. In some embodimentsThe provided stapled peptides are further modified and/or conjugated with other entities. The conditions and/or reagents of these reactions are well known in the art and may be performed in accordance with the present disclosure to provide a stapled peptide.
According to the present disclosure, the characteristics and/or activity of the provided stapled peptides can be readily assessed, for example, by using one or more of the methods described in the examples.
In some embodiments, techniques for preparing and/or evaluating the provided stapled peptides include those described in U.S. Pat. No. 4,9617309, U.S. Pat. No. 5,022,5471, U.S. Pat. No. 2016-0024153, U.S. Pat. No. 5,021 5036, U.S. Pat. No. 2016-024494, WO 2017/062518, and the like.
In some embodiments, the present disclosure provides products manufactured and/or characterized by the processes and/or techniques described herein.
In some embodiments, the provided compounds, e.g., amino acids or protected forms thereof, can be prepared using the following techniques.
In some embodiments, one or more or all of the following steps may be used to prepare the provided compounds:
those skilled in the art will appreciate that other leaving groups (e.g., -Br, -I, -OTs, oms, etc.) may be used in place of-Cl for the first reaction.
In some embodiments, one or more or all of the following steps may be used to prepare the provided compounds:
in some embodiments, one or more or all of the following steps may be used to prepare the provided compounds:
In some embodiments, one or more or all of the following steps may be used to prepare the provided compounds:
in some embodiments, one or more or all of the following steps may be used to prepare the provided compounds:
the provided compounds can be provided in high purity. In some embodiments, the provided compounds are at least about 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% pure. In some embodiments, the provided compounds (e.g., optionally protected/activated amino acids) are substantially free of impurities, including stereoisomers.
In some embodiments, the pharmaceutical agent may have one or more stereoisomers that may independently coexist in the composition or formulation. In some embodiments, the provided pharmaceutical agent has a stereoscopic purity of about or at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, it is about or at least about 80%. In some embodiments, it is about or at least about 85%. In some embodiments, it is about or at least about 90%. In some embodiments, it is about or at least about 95%. In some embodiments, it is about or at least about 96%. In some embodiments, it is about or at least about 97%. In some embodiments, it is about or at least about 98%. In some embodiments, it is about or at least about 99%. In some embodiments, the formulation or composition is substantially free of stereoisomers (e.g., not reliably observed in NMR or HPLC). In some embodiments, the medicament comprises one or more staple-like structures independently containing one or more olefinic double bonds. In some embodiments, the stereochemistry is with respect to the E/Z stereoisomer. In some embodiments, the ratio of the two stereoisomers is independently about 1.1:1, 1.2: 1,1.3: 1,1.4:1, 1.5:1, 2:1, 3:1, 4:1, 5:1, 10:1, 20:1, 30:1, 40:1, 50:1, 60:1, 70:1, 80:1, 90:1, 100:1 or higher for one or more (e.g., 1,2, 3, 4, 5 or more) of the staple-like structures or each olefinic double bond. In some embodiments, independently, it is about 1.5:1 or higher. In some embodiments, independently, it is about 2:1 or higher. In some embodiments, independently, it is about 3:1 or higher. In some embodiments, independently, it is about 4:1 or higher. In some embodiments, independently, it is about 5:1 or higher. In some embodiments, independently, it is about 6:1 or higher. In some embodiments, independently, it is about 7:1 or higher. In some embodiments, independently, it is about 8:1 or higher. In some embodiments, it is independently about 9:1 or higher. In some embodiments, independently, it is about 10:1 or higher. In some embodiments, it is independently about 20:1 or higher. In some embodiments, independently, it is about 30:1 or higher. In some embodiments, it is independently about 40:1 or higher. In some embodiments, independently, it is about 50:1 or higher. In some embodiments, independently, it is about 60:1 or higher. In some embodiments, independently, it is about 70:1 or higher. In some embodiments, it is independently about 80:1 or higher. In some embodiments, it is independently about 90:1 or higher. In some embodiments, independently, it is about 100:1 or higher. In some embodiments, the ratio is E:Z. In some embodiments, the ratio is Z: E. Those skilled in the art will appreciate that the E and Z isomers may be selectively enriched by adjusting the manufacturing process, purification, staple structure location and/or length, and the like.
Composition and method for producing the same
The present disclosure provides, inter alia, compositions comprising or otherwise related to: agents provided as described herein, e.g., small molecule agents, peptide agents (e.g., stapled peptides).
In some embodiments, provided compositions are or comprise the following: an assay system for characterizing (and optionally comprising) a stapled peptide as described herein.
In some embodiments, the provided compositions are pharmaceutical compositions, e.g., comprising or delivering one or more provided agents.
In some embodiments, the agent is a peptide. In some embodiments, the agent is a stapled peptide. In some embodiments, the agent comprises a detectable moiety, e.g., a fluorescent moiety, a radioactive moiety, biotin, and the like. In some embodiments, the detectable moiety is directly detectable. In some embodiments, the detectable antibody is indirectly detectable, e.g., using an antibody, agent, etc., that can react with the detectable moiety to form a detectable product.
In some embodiments, the pharmaceutical composition comprises the provided agent, and a pharmaceutically acceptable excipient (e.g., carrier).
In some embodiments, the peptide composition can comprise or deliver a particular form (e.g., a particular optical isomer, diastereomer, salt form, covalent conjugate form [ e.g., covalently linked to a carrier moiety ], etc., or a combination thereof) of an agent as described herein. In some embodiments, described herein are pharmaceutical compositions comprising or delivered by an agent that is not covalently linked to a carrier moiety.
In some embodiments, the agent comprising a chiral center and/or a double bond is present in a plurality of stereoisomers. In some embodiments, the level of a particular agent in the composition is enriched relative to one or more or all of its stereoisomers. For example, in some embodiments, the specific configuration of the double bond (E/Z) is enriched. In some embodiments, the configuration is enriched independently for each double bond. In some embodiments, one configuration is enriched for chiral elements, such as chiral centers. In some embodiments, one configuration is enriched for chiral centers bonded to two staple-like structures. In some embodiments, the configuration is enriched independently for each chiral element. In some embodiments, one configuration is independently enriched for one or more or all stereochemical elements (e.g., double bonds, chiral elements, etc.). In some embodiments, one configuration is enriched independently for each double bond in each staple-like structure. In some embodiments, one configuration is enriched independently for each double bond in each staple-like structure, and one configuration is enriched for chiral centers bonded to both staple-like structures. In some embodiments, the enrichment of each double bond is independently E or Z. In some embodiments, the enrichment of each chiral element is independently R or S. In some embodiments, the enrichment of each stereochemical element (e.g., double bond, chiral center, etc.) is about or at least about a level, such as 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% (percent of agent). In some embodiments, about or at least about a level of all molecules of the composition of the shared agent or salt thereof in the composition, e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% is the agent or salt thereof. In some embodiments, the level is about or at least about 60%. In some embodiments, it is about or at least about 65%. In some embodiments, it is about or at least about 70%. In some embodiments, it is about or at least about 75%. In some embodiments, it is about or at least about 80%. In some embodiments, it is about or at least about 85%. In some embodiments, it is about or at least about 90%. In some embodiments, it is about or at least about 95%. In some embodiments, it is about or at least about 96%. In some embodiments, it is about or at least about 97%. In some embodiments, it is about or at least about 98%. In some embodiments, it is about or at least about 99%.
In some embodiments, provided therapeutic compositions may comprise one or more additional therapeutic agents and/or one or more stabilizing agents and/or one or more agents that alter (e.g., extend to or limit the rate or extent of delivery to a particular tissue, location or site) over time.
In some embodiments, the composition is a pharmaceutical composition comprising or delivering the provided agent (e.g., a stapled peptide) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. In some embodiments, the composition comprises one and only stereoisomers of the pharmaceutical agent (e.g., a stapled peptide) and/or one or more salts thereof. In some embodiments, the composition comprises two or more stereoisomers of the agent (e.g., a stapled peptide) and/or one or more salts thereof. In some embodiments, two or more stereoisomers of the agent (e.g., stapled peptide) or salt thereof elute as a single peak (e.g., UV and/or MS detection) in a chromatograph (e.g., HPLC).
Use and application
The agents and compositions provided may be used for a variety of purposes. For example, certain compounds may be used directly as amino acids, or may be used as amino acids for the preparation of other compounds, such as peptides. Certain agents (e.g., peptides) may be used to prepare the stapled peptides. Certain agents, and compositions thereof, that are or comprise peptides, particularly stapled peptides, are biologically active and useful for a variety of purposes, e.g., as therapeutic agents against a variety of conditions, disorders, or diseases, as a means for modulating biological function, and the like.
In some embodiments, the present disclosure provides agents and compositions thereof for modulating β -catenin function. In some cases, β -catenin is reported to have a variety of cellular functions, including intercellular adhesion and regulation and coordination of gene transcription. In some embodiments, the agents described herein may inhibit β -catenin activity and/or levels, and may, for example, inhibit tumor growth. In some embodiments, the agents described herein can activate and/or increase the level of β -catenin, and can be used, for example, to treat male pattern baldness or hair loss.
Beta-catenin has been reported to interact with TCF/LEF family members at the TCF site on beta-catenin. In some embodiments, the provided techniques may reduce, inhibit, or block one or more such interactions. In some embodiments, the present disclosure provides methods for modulating the interaction between β -catenin and its binding partner (e.g., a TCF/LEF family member), comprising contacting β -catenin with a provided agent.
In some embodiments, the provided agent competes with or inhibits binding of the additional agent to β -catenin. In some embodiments, the provided agent competes with or inhibits binding of the additional agent to β -catenin. In some embodiments, the provided agents compete with or inhibit the binding of TCF or a fragment thereof to β -catenin.
In some embodiments, the provided agents compete for β -catenin binding with TCF7, LEF1, TCF7L2, axin1, axin2, APC, CDH1, or CDH2, or fragments thereof.
In some embodiments, the provided agent interferes with the interaction of TCF7, LEF1, TCF7L2, axin1, axin2, APC, CDH1, or CDH2, or fragments thereof, with β -catenin.
In some embodiments, the provided techniques can reduce or block the interaction of β -catenin with all TCF family members, E-cadherins, and APCs, but do not significantly affect their interaction with ICAT, AXIN, and BCL 9. In some embodiments, the provided techniques can interrupt β -catenin/TCF interactions at both physical interaction levels (e.g., as determined by NanoBRET, co-IP, etc.) and transcriptional levels (e.g., as determined by a reporter cell line, endogenous gene expression, etc.). In some embodiments, the provided techniques exhibit no effect on β -catenin stability.
In some embodiments, the disclosure provides methods for modulating the interaction of β -catenin with a partner (e.g., TCF7, LEF1, TCF7L2, axin1, axin2, APC, CDH1, or CDH2, or fragments thereof), the methods comprising contacting β -catenin with a provided agent or a composition comprising or delivering a provided agent. In some embodiments, the disclosure provides methods for modulating the interaction of β -catenin with a partner (e.g., TCF7, LEF1, TCF7L2, axin1, axin2, APC, CDH1, or CDH2, or fragments thereof) comprising administering or delivering a provided agent or a composition comprising a provided agent or delivering a provided agent to a system comprising β -catenin and a partner. In some embodiments, the system is an in vitro system. In some embodiments, the system is an in vivo system. In some embodiments, the system is or comprises a cell, tissue or organ. In some embodiments, the system is an object. In some embodiments, the present disclosure provides methods for inhibiting cell growth comprising administering or delivering to a population of cells an effective amount of the provided agents or pharmaceutically acceptable salts thereof. In some embodiments, the present disclosure provides methods for killing cells associated with a condition, disorder, or disease (e.g., cancer), the methods comprising administering or delivering an effective amount of a provided agent, or a pharmaceutically acceptable salt thereof, to such cell populations.
In some embodiments, the present disclosure provides methods for preventing a condition, disorder, or disease associated with β -catenin (e.g., cancer, neurodegenerative disease, etc.), the methods comprising administering or delivering an effective amount of the provided agents, or pharmaceutically acceptable salts thereof, to a subject susceptible to the condition, disorder, or disease. In some embodiments, the present disclosure provides methods for treating a condition, disorder, or disease associated with β -catenin (e.g., aberrant β -catenin activity and/or expression levels), comprising administering or delivering an effective amount of a provided agent, or a pharmaceutically acceptable salt thereof, to a subject suffering from the condition, disorder, or disease. In some embodiments, the provided agent is administered as a pharmaceutical composition comprising or delivering an effective amount of the provided agent or a pharmaceutically acceptable salt thereof. In some embodiments, the condition, disorder or disease and the interaction of β -catenin with a partner (e.g., TCF7, LEF1, TCF7L2, axin1, axin2, APC, CDH1, and/or CDH 2). In some embodiments, the condition, disorder or disease and β -catenin are associated with TCF. In some embodiments, the condition, disorder or disease is cancer. In some embodiments, the provided agents can be administered in combination with additional therapies (e.g., immunotherapy). In some embodiments, the condition, disorder or disease is selected from the group consisting of cancer, heart disease, dilated cardiomyopathy, fetal alcohol syndrome, depression, and diabetes. In some embodiments, the condition, disorder or disease is a cardiac condition, disorder or disease. In some embodiments, the condition, disorder or disease is cancer. In some embodiments, the cancer is selected from: colon cancer, colorectal cancer, rectal cancer, prostate cancer, familial adenomatous polyposis (familial adenomatous polyposis, FAP), wilms Tumor, melanoma, hepatocellular carcinoma, ovarian cancer, endometrial cancer, medulloblastoma (medullobastoma), medulloblastoma (pilomastia), primary hepatocellular carcinoma, ovarian cancer, breast cancer, lung cancer, glioblastoma, medullomastia (pliomastia), medulloblastoma, thyroid Tumor, and ovarian neoplasms. In some embodiments, the condition, disorder or disease is cancer, such as colorectal cancer, hepatocellular carcinoma, melanoma, gastric cancer, bladder cancer, and endometrial cancer. In some embodiments, the cancer is colorectal cancer. In some embodiments, the cancer is hepatocellular carcinoma. In some embodiments, the cancer is prostate cancer. In some embodiments, the cancer is melanoma.
In some embodiments, the present disclosure provides techniques for modulating the expression and/or activity level of a nucleic acid (e.g., a gene), transcript, polypeptide, and/or product thereof in a system comprising administering or delivering a provided agent or a composition comprising or delivering a provided agent to a system. In some embodiments, the expression level of a nucleic acid (e.g., gene) or a product thereof (e.g., transcript, polypeptide, etc.) is modulated. In some embodiments, the level of activity of a nucleic acid (e.g., gene) or a product thereof (e.g., transcript, polypeptide, etc.) is modulated. In some embodiments, the level of the transcript and/or product thereof (e.g., polypeptide) is modulated. In some embodiments, the level of activity of the transcript and/or product thereof (e.g., polypeptide) is modulated. In some embodiments, the transcript is a transcript of a nucleic acid (e.g., a gene) as described herein. In some embodiments, the level of the polypeptide is modulated. In some embodiments, the level of activity of the polypeptide is modulated. In some embodiments, the polypeptide is encoded by a nucleic acid or transcript as described herein. In some embodiments, the level is increased. In some embodiments, the level is reduced. In some embodiments, the system is an in vitro system, as described herein. In some embodiments, the system is an in vivo system. In some embodiments, the system is or comprises a cell, tissue or organ. In some embodiments, the system is or comprises one or more cancer cells. In some embodiments, the system is or comprises a tumor. In some embodiments, the system is or comprises an organism. In some embodiments, the system is an object. In some embodiments, the system is a human. In some embodiments, the system comprises β -catenin. In some embodiments, the system expresses β -catenin. In some embodiments, the system comprises β -catenin and a partner. In some embodiments, the system expresses β -catenin and a partner. In some embodiments, the level is modulated by β -catenin. In some embodiments, the level is modulated by WNT activation. In some embodiments, the level is modulated by β -catenin/WNT signaling. In some embodiments, the level is modulated by the interaction of β -catenin and the partner. In some embodiments, the interaction of β -catenin and a partner is modulated, e.g., reduced, prevented, etc., by an agent (e.g., a stapled peptide) as described herein. For example, in some embodiments, the partner is TCF. In some embodiments, the expression and/or activity level of the nucleic acid and/or product thereof is modulated. In some embodiments, the nucleic acid is AXIN2. In some embodiments, the level of AXIN2 transcript (e.g., mRNA) is reduced. In some embodiments, the level of an AXIN2 polypeptide is reduced. In some embodiments, the nucleic acid is SP5. In some embodiments, the level of SP5 transcript (e.g., mRNA) is reduced. In some embodiments, the level of SP5 polypeptide is reduced. In some embodiments, the nucleic acid is CXCL12. In some embodiments, the level of CXCL12 transcript (e.g., mRNA) is increased. In some embodiments, the level of CXCL12 polypeptide is increased. In some embodiments, the nucleic acid is a member of a negatively enriched genome as observed, for example, in example 17, or identifiable using, for example, the techniques in example 17. In some embodiments, the nucleic acid is a member of the bcat_gds748_up genome. In some embodiments, the nucleic acid is a member of the bcat.100_up.v1_up genome. In some embodiments, the nucleic acid is a member of the hallmark_wnt_beta_catenin_signaling genome. In some embodiments, the nucleic acid is a member of the rashi_response_to_ionizingjradiation_1 genome. In some embodiments, the nucleic acid is a member of the REACTOME_RRNA_PROCESSING genome. In some embodiments, the nucleic acid is a member of the hallmark_myc_target_v1 genome. In some embodiments, the nucleic acid is a member of the hallmark_myc_target_v2 genome. In some embodiments, the nucleic acid is a member of the hallmark_oxidation_PHOSPHORYLATION genome. In some embodiments, the nucleic acid is a member of the hallmark_e2f_target genome. In some embodiments, the nucleic acid is a member of the hallmark_tnfa_signature_via_nfkb genome. Descriptions of various genomes can be found publicly, e.g., https:// www.gsea-msigdb. In some embodiments, one or more or some or most but not all of the nucleic acids or genes in the genome are affected in the same manner, but the genome as a whole may be negatively or positively enriched. In some embodiments, the nucleic acid is selected from table GS1. In some embodiments, the nucleic acid is selected from table GS2. In some embodiments, the nucleic acid is selected from GS3. In some embodiments, the nucleic acid is selected from table GS4. In some embodiments, the nucleic acid is selected from table GS5. In some embodiments, the nucleic acid is selected from table GS6. In some embodiments, the nucleic acid is selected from table GS7. In some embodiments, the nucleic acid is selected from table GS8. In some embodiments, the nucleic acid is selected from table GS9. In some embodiments, the nucleic acid is selected from table GS10. In some embodiments, the nucleic acid is a gene selected from table GS1, table GS2, table GS3, table GS4, table GS5, table GS6, table GS7, table GS8, table GS9, or table GS10. In some embodiments, the gene is CCND2. In some embodiments, the gene is WNT5B. In some embodiments, the gene is AXIN2. In some embodiments, the gene is NKD1. In some embodiments, the gene is WNT6. In some embodiments, the gene is DKK1. In some embodiments, the gene is DKK4. In some embodiments, the expression of such nucleic acids (e.g., genes) is reduced. In some embodiments, the level of a product (e.g., transcript (e.g., mRNA), polypeptide, etc.) of such nucleic acids is reduced. In some embodiments, the level of activity of a product (e.g., transcript (e.g., mRNA), polypeptide, etc.) of such a nucleic acid is reduced.
Table gs1. Some examples of nucleic acids comprising a plurality of members of bcat_gds748_up.
Table gs2. Some examples of nucleic acids comprising a plurality of members of bcat.100_up.v1_up.
Table GS3 some examples of nucleic acids comprising a plurality of members of HALMARK_WNT_BETA_CATENIN_SIGNALING.
Table GS4 some examples of nucleic acids comprising a plurality of members of RASHI_RESPONSE_TO_IONIZING_RADIATION_1.
Table GS5 some examples of nucleic acids comprising members of REACTOME_RRNA_PROCESSING.
Table GS6 some examples of nucleic acids comprising a plurality of members of HALMARK_MYC_TARGETS_V1.
Table GS7 some examples of nucleic acids comprising a plurality of members of HALMARK_MYC_TARGETS_V2.
Table GS8 some examples of nucleic acids comprising members of hallmark_oxidation_PHOSPHORYLATION.
Table GS9 some examples of nucleic acids comprising a plurality of members of HALMARK_E2F_TARGETS.
Table GS10 some examples of nucleic acids comprising a plurality of members of HALMARK_TNFA_SIGNALING_VIA_NFKB.
In some embodiments, the nucleic acid is a member of a positive enriched genome as observed, for example, in example 17, or identifiable using, for example, the techniques in example 17.
In some embodiments, the present disclosure provides techniques for detecting, monitoring, and/or determining the efficacy of an agent (e.g., a stapled peptide) or method (e.g., a method of treating a condition, disorder, or disease, a method of modulating the level of a transcript and/or product and/or activity thereof) comprising assessing the level of expression and/or activity of a nucleic acid (e.g., a gene), transcript, polypeptide, and/or product thereof. In some embodiments, the present disclosure provides techniques for detecting, monitoring, and/or determining the efficacy of an agent (e.g., a stapled peptide) comprising administering the agent to a subject and assessing the expression and/or activity level of a nucleic acid (e.g., a gene), transcript, polypeptide, and/or product thereof in the subject. In some embodiments, the present disclosure provides techniques for detecting, monitoring, and/or determining the efficacy of a method for treating a condition, disorder, or disease in a subject, comprising assessing the expression and/or activity level of a nucleic acid (e.g., gene), transcript, polypeptide, and/or product thereof in the subject. In some embodiments, the method is a method for treating a condition, disorder or disease associated with TCF- β -catenin interactions in a subject. In some embodiments, the condition, disorder or disease is cancer as described herein. In some embodiments, the present disclosure provides techniques for selecting a subject for administration or delivery of an agent (e.g., a stapled peptide agent) described herein (e.g., for preventing or treating a condition, disorder, or disease). In some embodiments, the present disclosure provides techniques for selecting a subject to continue administration or delivery of an agent (e.g., a stapled peptide agent) described herein (e.g., for preventing or treating a condition, disorder, or disease) after one or more administrations or deliveries. In some embodiments, the level of the transcript is assessed. In some embodiments, the level of the polypeptide is assessed. In some embodiments, the evaluation is performed using one or more samples collected from the system or object. In some embodiments, the sample is collected during administration or delivery. In some embodiments, the sample is collected after administration or delivery. As described herein, in some embodiments, the expression and/or activity level of a nucleic acid and/or product thereof is modulated. In some embodiments, the nucleic acid is AXIN2. In some embodiments, the level of AXIN2 transcript (e.g., mRNA) is reduced. In some embodiments, the level of an AXIN2 polypeptide is reduced. In some embodiments, the nucleic acid is SP5. In some embodiments, the level of SP5 transcript (e.g., mRNA) is reduced. In some embodiments, the level of SP5 polypeptide is reduced. In some embodiments, the nucleic acid is CXCL12. In some embodiments, the level of CXCL12 transcript (e.g., mRNA) is increased. In some embodiments, the level of CXCL12 polypeptide is increased. In some embodiments, the nucleic acid is a member of a negatively enriched genome as observed, for example, in example 17, or identifiable using, for example, the techniques in example 17. In some embodiments, the nucleic acid is a member of the bcat_gds748_up genome. In some embodiments, the nucleic acid is a member of the bcat.100_up.v1_up genome. In some embodiments, the nucleic acid is a member of the hallmark_wnt_beta_catenin_signaling genome. In some embodiments, the nucleic acid is a member of the rashi_response_to_ionizingjradiation_1 genome. In some embodiments, the nucleic acid is a member of the REACTOME_RRNA_PROCESSING genome. In some embodiments, the nucleic acid is a member of the hallmark_myc_target_v1 genome. In some embodiments, the nucleic acid is a member of the hallmark_myc_target_v2 genome. In some embodiments, the nucleic acid is a member of the hallmark_oxidation_PHOSPHORYLATION genome. In some embodiments, the nucleic acid is a member of the hallmark_e2f_target genome. In some embodiments, the nucleic acid is a member of the hallmark_tnfa_signature_via_nfkb genome. In some embodiments, the nucleic acid is selected from table GS1. In some embodiments, the nucleic acid is selected from table GS2. In some embodiments, the nucleic acid is selected from table GS3. In some embodiments, the nucleic acid is selected from table GS4. In some embodiments, the nucleic acid is selected from table GS5. In some embodiments, the nucleic acid is selected from table GS6. In some embodiments, the nucleic acid is selected from table GS7. In some embodiments, the nucleic acid is selected from table GS8. In some embodiments, the nucleic acid is selected from table GS9. In some embodiments, the nucleic acid is selected from table GS10. In some embodiments, the nucleic acid is a gene selected from table GS1, table GS2, table GS3, table GS4, table GS5, table GS6, table GS7, table GS8, table GS9, or table GS10. In some embodiments, the gene is CCND2. In some embodiments, the gene is WNT5B. In some embodiments, the gene is AXIN2. In some embodiments, the gene is NKD1. In some embodiments, the gene is WNT6. In some embodiments, the gene is DKK1. In some embodiments, the gene is DKK4. In some embodiments, the expression of such nucleic acids (e.g., genes) is reduced. In some embodiments, the level of a product (e.g., transcript (e.g., mRNA), polypeptide, etc.) of such nucleic acids is reduced. In some embodiments, the level of activity of a product (e.g., transcript (e.g., mRNA), polypeptide, etc.) of such a nucleic acid is reduced. In some embodiments, the nucleic acid is a member of a positive enriched genome as observed, for example, in example 17, or identifiable using, for example, the techniques in example 17. In some embodiments, if one or more desired decreases in expression and/or levels of the transcript and/or product thereof, and/or one or more desired negative and/or positive enriched genomes are observed, administration or delivery is continued. In some embodiments, administration or delivery continues as before. In some embodiments, administration or delivery is continued at the adjusted dosage level and/or regimen. In some embodiments, administration or delivery may be adjusted if no desired reduction in expression and/or level of the transcript and/or its product, and/or one or more desired negative and/or positive enriched genomes is observed, and in some embodiments, administration or delivery may be stopped. In some embodiments, as described herein, the desired reduction in expression and/or level of a transcript and/or product thereof comprises a reduction in expression and/or level of one or more or most or all of the following transcripts and/or products thereof: SP5, CCND2, WNT5B, AXIN2, NKD1, WNT6, DKK1 and DKK4, bcat_gds748_up, bcat.100_up.v1_up, hallmark_wnt_BETA_CATENIN_ SIGNALING, RASHI _response_to_ionZING_radio_1, reactome_rrna_process, hallmark_myc_target_v1, hallmark_myc_target_v2, hallmark_target_v_2, hallmark_oxidation_phosphate_target, hallmark_e2f_target_s, hallmark_tnfa_sign_nfkb and nucleic acids of table GS1, table GS2, table GS3, table GS4, table GS5, table GS6, table 7, table GS8, table GS9 and table GS10. In some embodiments, a desired increase in the expression and/or level of a transcript and/or product thereof as described herein comprises an increase in the expression and/or level of a transcript and/or product thereof of CXCL12. In some embodiments, the desired genomic enrichment comprises negative enrichment of one or more or all of the following: bcat_gds748_up, bcat.100_up.v1_up, hallmark_wnt_beta_catin_ SIGNALING, RASHI _response_to_ioniz_radiation_1, REACTOME_rrna_process_signal, hallmark_myc_target_v1, hallmark_myc_target_v2, hallmark_oxide_phosphate_target, hallmark_e2f_target_s, and hallmark_tnfa_signal_vig_nfkb. In some embodiments, the desired genomic enrichment comprises negative enrichment of one or more or all of the following: the group in table GS1, the group in table GS2, the group in table GS3, the group in table GS4, the group in table GS5, the group in table GS6, the group in table GS7, the group in table GS8, the group in table GS9, and the group in table GS10. Those skilled in the art who review this disclosure will understand how to make decisions in accordance with this disclosure, for example, to those of skill in the relevant clinical arts.
In some embodiments, the comparison is made against a reference. For example, decreasing, increasing, enriching (negative or positive), changing, etc. are typically performed with respect to a suitable reference. In some embodiments, decreasing, increasing, enriching (negative or positive), altering, etc. are in some embodiments assessed against a reference of the reference sample. In some embodiments, the reference assessment is or includes the following: evaluation prior to administration or delivery of the agent. In some embodiments, the reference sample is collected prior to administration or delivery of the agent. In some embodiments, the reference assessment is or includes the following: evaluation during administration or delivery of the agent. In some embodiments, the reference sample is collected during administration or delivery of the agent. In some embodiments, the reference assessment is or includes the following: evaluation performed after administration or delivery of the agent. In some embodiments, the reference sample is collected after administration or delivery of the agent. In some embodiments, the reference assessment is or includes the following: evaluation performed after an earlier administration or delivery of the agent. In some embodiments, the reference sample is collected after an earlier administration or delivery of the agent.
In some embodiments, the sample is an aliquot of a substance obtained from or derived from a source of interest as described herein. In some embodiments, the source of interest is a biological or environmental source. In some embodiments, the source of interest may be or include the following: a cell or organism, such as a microorganism, a plant or an animal (e.g., a human). In some embodiments, the source of interest is or comprises biological tissue or fluid. In some embodiments, the biological tissue or fluid may be or comprise the following: amniotic fluid, aqueous humor, ascites, bile, bone marrow, blood, breast milk, cerebrospinal fluid, cerumen, chyle, chime, jet, endolymph (endomph), exudate, faeces, gastric acid, gastric fluid, lymph, mucus, pericardial fluid, perilymph (perilymph), peritoneal fluid, pleural fluid, pus, thin mucus (rheum), saliva, sebum, semen, serum, vaginal scale (smegma), sputum, synovial fluid, sweat, tears, urine, vaginal secretions, vitreous (vitreus) vomit, and/or combinations or components thereof. In some embodiments, the biological fluid may be or comprise the following: intracellular fluid, extracellular fluid, intravascular fluid (plasma), interstitial fluid, lymph fluid, and/or transcellular fluid. In some embodiments, the biological fluid may be or comprise plant exudates. In some embodiments, the biological tissue or sample may be obtained, for example, by aspiration, biopsy (e.g., fine needle or tissue biopsy), swab (e.g., oral, nasal, skin, or vaginal swab), scraping, surgery, washing, or lavage (e.g., brocheoalvealar, catheter, nose, eye, oral, uterus, vagina, or other washing or lavage). In some embodiments, the biological sample is or comprises cells obtained from an individual. In some embodiments, the sample is a "starting sample" obtained directly from the source of interest by any suitable means. In some embodiments, as will be clear from the context, the term "sample" refers to a preparation obtained by processing an initial sample (e.g., by removing one or more components of the initial sample and/or by adding one or more agents to the initial sample). For example, filtration using a semipermeable membrane. Such "treated sample" may comprise, for example, nucleic acids or proteins extracted from the sample, or obtained by subjecting the initial sample to one or more techniques (e.g., amplification or reverse transcription of nucleic acids, isolation and/or purification of certain components, etc.). In some embodiments, the sample comprises cancer cells. In some embodiments, the sample is obtained from a tumor. In some embodiments, the sample is obtained from a tumor of the patient.
In some embodiments, the level of two or more transcripts and/or products thereof may be assessed. In some embodiments, the evaluation is performed after administering or delivering one or more doses of the agent (e.g., the stapled peptide) to the subject. In some embodiments, if the profile (e.g., decrease, increase, etc.) of one or more transcripts and/or products thereof matches those described herein, administration or delivery to the subject may continue. In some embodiments, if the profile (e.g., decrease, increase, etc.) of one or more transcripts and/or products thereof matches those described herein, administration or delivery to a subject may be stopped and/or continued according to different dosage levels and/or regimens.
In accordance with the present disclosure, the provided technology, e.g., agents, peptides, compounds, compositions, and the like, may be formulated, distributed, administered, or delivered using a variety of techniques. For example, in some embodiments, administration may be ocular, oral, parenteral, topical, and the like. In some embodiments, administration may be transbronchial (e.g., by bronchial instillation), buccal (buccal), transdermal (which may be or include, e.g., surface to one or more of dermal, intradermal (inter-dermal), transdermal, etc.), enteral, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular (intra-ocular), within a particular organ (e.g., intrahepatic), transmucosal, nasal, oral, rectal, subcutaneous, sublingual, surface, tracheal (e.g., by intratracheal instillation), vaginal, vitreous, etc. In some embodiments, administration may involve intermittent administration (e.g., multiple doses separated in time) and/or periodic administration (e.g., separate doses separated by a common period). In some embodiments, administration may involve continuous administration (e.g., infusion) for at least a selected period of time. In some embodiments, the provided technique is intravenous administration.
The present disclosure provides, inter alia, a variety of moieties including engineered amino acid residues that can be used to optimize a variety of properties and activities, stability, delivery, pharmacodynamics, pharmacokinetics, etc., to provide a variety of dosage forms, dosage regimens, therapeutic windows, etc. In some embodiments, the provided agents and compositions thereof may be used with improved dosage regimens and/or unit doses. In some embodiments, the administration of the provided agents is adjusted based on the condition, disorder, or disease and/or subpopulation. In some embodiments, the administration and/or dosage regimen of the provided technology is adjusted according to certain biomarkers and genomic changes.
The provided agents may deliver biological effects, such as therapeutic effects, through a variety of mechanisms. In some embodiments, efficacy may be driven by AUC. In some embodiments, potency may be driven by Cmax.
In some embodiments, the provided agents are used in combination with additional therapies. In some embodiments, the provided agents are used in combination with additional therapeutic agents. In some embodiments, the additional therapeutic or therapeutic agent is administered prior to administration or delivery of the provided agent. In some embodiments, the additional therapeutic or therapeutic agent is administered at about the same time as the provided agent is administered or delivered. In some embodiments, the agent provided and the additional agent are in the same pharmaceutical composition. In some embodiments, the additional therapeutic or therapeutic agent is administered after administration or delivery of the provided agent. In some embodiments, the subject is exposed to both the provided agent and the additional therapeutic agent. In some embodiments, both the provided agent and the additional agent may be detected in the subject. In some embodiments, the provided agent is administered before the additional agent is cleared by the subject, and vice versa. In some embodiments, the provided agent is administered within the half-life of the additional agent or within 2, 3, 4, 5, or 6 times the half-life, or vice versa. In some embodiments, the subject is exposed to the therapeutic effect of the provided agent and the therapeutic effect of the additional therapeutic agent. In some embodiments, the agent may provide an effect after the agent is cleared or metabolized by the subject. In some embodiments, a procedure (e.g., surgery, radiation, etc.) may provide an effect after the procedure is completed.
In some embodiments, the additional treatment is cancer treatment. In some embodiments, the additional treatment is or includes surgery. In some embodiments, the additional treatment is or comprises radiation therapy. In some embodiments, the additional treatment is or comprises an immunotherapy. In some embodiments, the additional therapeutic agent is or comprises a drug. In some embodiments, the additional therapeutic agent is or comprises a cancer drug. In some embodiments, the additional therapeutic agent is or comprises a chemotherapeutic agent. In some embodiments, the additional therapeutic agent is or comprises a hormonal therapeutic agent. In some embodiments, the additional therapeutic agent is or comprises a kinase inhibitor. In some embodiments, the additional therapeutic agent is or comprises a checkpoint inhibitor (e.g., an antibody to PD-1, PD-L1, CTLA-4, etc.). In some embodiments, the provided agents may be administered in lower unit doses and/or total doses than used alone. In some embodiments, the additional agent may be administered in a lower unit dose and/or total dose than if used alone. In some embodiments, one or more side effects associated with the provided agent and/or administration of additional therapeutic or therapeutic agents are reduced. In some embodiments, the combination therapy provides improved results, for example, when compared to each agent used alone. In some embodiments, the combination therapy achieves one or more better results, such as when compared to each agent used alone.
In some embodiments, the additional agent is a checkpoint inhibitor, an EGFR inhibitor, a VEGF inhibitor, a VEGFR inhibitor, a kinase inhibitor, or an anticancer drug.
In some embodiments, the additional agent is a checkpoint inhibitor. In some embodiments, the additional agent is an immunooncology agent. In some embodiments, the additional agent is an antibody to a checkpoint molecule. In some embodiments, the additional agent is an antibody to PD1, PDL-1, CTLA4, A2AR, B7-H3, B7-H4, BTLA, IDO, KIR, LAG3, TIM-s, C10orf54, or the like. In some embodiments, the antibody is an anti-PD 1 antibody. In some embodiments, the antibody is an anti-PD-L1 antibody. In some embodiments, the antibody is anti-CTLA 4.
In some embodiments, the additional agent is an EGFR inhibitor, e.g., erlotinib, gefitinib, lapatinib, panitumumab, vandetanib (vanretanib), cetuximab (cetuximab), and the like. In some embodiments, the additional agent is a VEGF and/or VEGFR inhibitor, e.g., pazopanib (pazopanib), bevacizumab (bevacizumab), sorafenib (sorafenib), sunitinib (sunitinib), axitinib (axitinib), panatinib (ponatinib), regorafenib (regorafenib), vandetanib (vanretetanib), cabozantinib (cabozantinib), ramucirumab (ramucirumab), lenvatinib, ziv-abamectin (ziv-aflibercept), and the like. In some embodiments, the additional agent is a kinase inhibitor. In some embodiments, the additional therapeutic agent is a chemotherapeutic agent. In some embodiments, the additional therapeutic agent is an anticancer agent, e.g., cyclophosphamide, methotrexate (methotrexate), 5-fluorouracil (5-fluorouracil, 5-FU), doxorubicin (doxorubicin), nitrogen mustard (mustine), vincristine (vincristine), procarbazine (procarbazine), prednisolone (prednisolone), dacarbazine (dacarbazine), bleomycin (bleomycin), etoposide (etoposide), cisplatin (cisplatin), epirubicin (epirubicin), capecitabine (capecitabine), leucovorin, actinomycin, all-trans retinoic acid, azacytidine (azacitidine), azathioprine (azathioprine), bortezomib (bortezomib), carboplatin (carboplatin), chlorambucil (chlorombil), cytarabine (cytarabine), daunorubicin (daunorubicin) docetaxel, doxifluridine, fluorouracil, gemcitabine, hydroxyurea, idarubicin, vinblastine, vindesine, vinorelbine, mitoxantrone, paclitaxel, pemetrexed, teniposide, and thioguanine, oxaliplatin (oxaliplatin), and the like.
The present disclosure provides, inter alia, the following embodiments:
1. an agent having the structure of formula I:
R N -L P1 -L AA1 -L P2 -L AA2 -L P3 -L AA3 -L P4 -L AA4 -L P5 -L AA5 -L P6 -L AA6 -L P7 -R C ，
I
wherein:
R N is a peptide, an amino protecting group or R' -L RN -；
L P1 、L P2 、L P3 、L P4 、L P5 、L P6 And L P7 Each independently is L, wherein L P1 、L P2 、L P3 、L P4 、L P5 、L P6 And L P7 Comprising:
a first R 'group and a second R' group taken together to form-L s -, the-L s -bonding to an atom to which a first R 'group is attached and an atom to which a second R' group is attached; and
a third R 'group and a fourth R' group taken together to form-L s -, the-L s -bonding to an atom to which a third R 'group is attached and an atom to which a fourth R' group is attached;
each L s Independently is-L s1 -L s2 -L s3 -, each L s1 、L s2 And L s3 Independently is L;
L AA1 is an amino acid residue comprising a side chain containing an acidic or polar group;
L AA2 is an amino acid residue comprising a side chain containing an acidic or polar group;
L AA3 is an amino acid residue;
L AA4 is an amino acid residue comprising a side chain comprising an optionally substituted aromatic group;
L AA5 is an amino acid residue comprising a side chain comprising an optionally substituted aromatic group;
L AA6 is a bagAmino acid residues containing side chains containing optionally substituted aromatic groups;
R C is a peptide, a carboxyl protecting group, -L RC -R’，-O-L RC -R 'or-N (R') -L RC R’；
L RN And L RC Each independently is L;
each L is independently a covalent bond, or an optionally substituted divalent C 1 -C 25 Aliphatic or heteroaliphatic groups having from 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -，-Cy-,-O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-;
each-Cy-is independently an optionally substituted divalent 3-to 30-membered monocyclic, bicyclic, or polycyclic ring having 0 to 10 heteroatoms;
each R' is independently-L-R, -C (O) R, -CO 2 R, or-SO 2 R；
Each R is independently-H or an optionally substituted group selected from: c (C) 1-30 Aliphatic, C having 1 to 10 heteroatoms 1-30 Heteroaliphatic, C 6-30 Aryl, C 6-30 Arylaliphatic, C having 1 to 10 heteroatoms 6-30 Aryl heteroaliphatics, 5 to 30 membered heteroaryl groups having 1 to 10 heteroatoms, and 3 to 30 membered heterocyclyl groups having 1 to 10 heteroatoms, or
The two R groups are optionally and independently taken together to form a covalent bond, or:
two or more R groups on the same atom optionally and independently join together with the atom to form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the atom; or alternatively
Two or more R groups on two or more atoms optionally and independently together with their intervening atoms form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the intervening atoms.
2. An agent having the structure of formula I:
R N -L P1 -L AA1 -L P2 -L AA2 -L P3 -L AA3 -L P4 -L AA4 -L P5 -L AA5 -L P6 -L AA6 -L P7 -R C ，
I
wherein:
R N is a peptide, an amino protecting group or R' -L RN -；
L P1 、L P2 、L P3 、L P4 、L P5 、L P6 And L P7 Each independently is L, wherein L P1 、L P2 、L P3 、L P4 、L P5 、L P6 And L P7 Comprising:
a first R 'group and a second R' group taken together to form-L s -, the-L s -bonding to an atom to which a first R 'group is attached and an atom to which a second R' group is attached; and
a third R 'group and a fourth R' group taken together to form-L s -, the-L s -bonding to an atom to which a third R 'group is attached and an atom to which a fourth R' group is attached;
each L s Independently is-L s1 -L s2 -L s3 -, each L s1 、L s2 And L s3 Independently is L;
L AA1 is L AR Wherein the methylene units are represented by-C (R') (R AS ) -substitution, wherein R AS is-L AS1 -R AA1 Wherein R is AA1 is-CO 2 R or-SO 2 R；
L AA2 Is L AR Wherein the methylene units are represented by-C (R') (R AS ) -substitution, wherein R AS is-L As2 -R AA2 Wherein R is AA2 is-CO 2 R or-SO 2 R；
L AA3 Is L AR Wherein the methylene units are represented by-C (R') (R AS ) -substitution, wherein R AS is-L AS3 -R AA3 Wherein R is AA3 Is R';
L AA4 is L AR Wherein the methylene units are represented by-C (R') (R AS ) -substitution, wherein R AS is-L AS4 -R AA4 Wherein R is AA4 Is an optionally substituted group selected from: a 6 to 14 membered aryl or a 5 to 14 membered heteroaryl having 1 to 6 heteroatoms;
L AA5 is L AR Wherein the methylene units are represented by-C (R') (R AS ) -substitution, wherein R AS is-L AS5 -R AA5 Wherein R is AA5 Is an optionally substituted group selected from: a 6 to 14 membered aryl or a 5 to 14 membered heteroaryl having 1 to 6 heteroatoms;
L AA6 is L AR Wherein the methylene units are represented by-C (R') (R AS ) -substitution, wherein R AS is-L AS6 -R AA6 Wherein R is AA6 Is an optionally substituted group selected from: a 6 to 14 membered aryl or a 5 to 14 membered heteroaryl having 1 to 6 heteroatoms;
R C is a peptide, a carboxyl protecting group, -L RC -R’，-O-L RC -R 'or-N (R') -L RC -R’；
L RN And L RC Each independently is L;
each L AR Independently an optionally substituted divalent C 1 -C 6 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -，-C(R’)(R AS )-，-Cy-，-O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 N (R') -C (O) S-, or-C (O) O-;
L AS1 、L As2 、L AS3 、L AS4 、L AS5 and L AS6 Each independently is L AS ；
Each R AS Independently is-L AS -R’；
Each L AS Independently a covalent bond, or an optionally substituted divalent C 1 To C 10 Aliphatic or heteroaliphatic groups having from 1 to 5 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -,-Cy-，-O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 N (R') -C (O) S-, or-C (O) O-;
each L is independently a covalent bond, or an optionally substituted divalent C 1 -C 25 Aliphatic or heteroaliphatic groups having from 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-;
each-Cy-is independently an optionally substituted divalent 3-to 30-membered monocyclic, bicyclic, or polycyclic ring having 0 to 10 heteroatoms;
each R' is independently-L-R, -C (O) R, -CO 2 R, or-SO 2 R；
Each R is independently-H or an optionally substituted group selected from: c (C) 1-30 Aliphatic, C having 1 to 10 heteroatoms 1-30 Heteroaliphatic, C 6-30 Aryl, C 6-30 Arylaliphatic, C having 1 to 10 heteroatoms 6-30 Aryl heteroaliphatics, 5 to 30 membered heteroaryl groups having 1 to 10 heteroatoms, and 3 to 30 membered heterocyclyl groups having 1 to 10 heteroatoms, or
The two R groups optionally and independently together form a covalent bond, or:
two or more R groups on the same atom optionally and independently taken together with the atom form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the atom; or alternatively
Two or more R groups on two or more atoms optionally and independently together with their intervening atoms form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the intervening atoms.
3. The agent of any of the preceding embodiments, wherein the second R 'group and the third R' group are attached to the same atom.
4. The agent of any one of the preceding embodiments, wherein the first R ' group, the second R ' group, and the fourth R ' group are each independently attached to a different atom.
5. The agent of any one of the preceding embodiments, wherein L P1 、L P2 、L P3 、L P4 、L P5 、L P6 And L P7 Also contains a fifth R 'group and a sixth R' group, which together form-L s -, the-L s -bonding to the atom to which the fifth R 'group is attached and to the atom to which the sixth R' group is attached.
6. The agent of any one of the preceding embodiments, wherein the first R ' group, the second R ' group, the fourth R ' group, the fifth R ' group, and the sixth R ' group are each independently attached to a different atom.
7. The agent of any one of the preceding embodiments, wherein L P1 、L P2 、L P3 、L P4 、L P5 、L P6 And L P7 Also contains a seventh R 'group and an eighth R' group, which together form-L s -, the-L s -bonding to the atom to which the seventh R 'group is attached and to the atom to which the eighth R' group is attached.
8. The agent of any one of the preceding embodiments, wherein the first R ' group, the second R ' group, the fourth R ' group, the fifth R ' group, the sixth R ' group, the seventh R ' group, and the eighth R ' group are each independently attached to a different atom.
9. The agent of any of the preceding embodiments, wherein L formed by the first R 'group and the second R' group taken together s Is a staple like structure as described herein.
10. The agent of any of the preceding embodiments, wherein L formed by the first R 'group and the second R' group taken together s Is 5 to 20 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) atoms in length.
11. The agent of any of the preceding embodiments, wherein L formed by the third R 'group taken together with the fourth R' group s Is a staple like structure as described herein.
12. The agent of any of the preceding embodiments, wherein L formed by the third R 'group taken together with the fourth R' group s Is 5 to 20 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) atoms in length.
13. The agent of any of the preceding embodiments, wherein L formed by the third R 'group taken together with the fourth R' group s Is 10 to 20 (e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) atoms in length.
14. The agent of any of the preceding embodiments, wherein L is formed by the fifth R 'group taken together with the sixth R' group s Is 5 to 20 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) atoms in length.
15. The agent of any of the preceding embodiments, wherein L formed by the seventh R 'group taken together with the eighth R' group s Is 5 to 20 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) atoms in length。
16. The agent of any one of the preceding embodiments, wherein L P1 Is a covalent bond or an optionally substituted divalent C 2 -C 6 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
17. The agent of any one of the preceding embodiments, wherein L P1 Is 2 to 10 (2, 3, 4, 5, 6, 7, 8, 9 or 10) atoms in length.
18. The agent of any one of the preceding embodiments, wherein L P1 Independently replaced by-N (R') -or-C (O) -.
19. The agent of any one of the preceding embodiments, wherein L P1 Independently replaced by-N (R ') -or-C (O) N (R') -.
20. The agent of any one of the preceding embodiments, wherein L P1 Is independently substituted with-N (R '), -C (R') 2 or-C (O) N (R') -substitution.
21. The agent of any one of the preceding embodiments, wherein L P1 Independently replaced by-N (R') -, and L P1 Independently replaced by-C (O) N (R') -.
22. The agent of any one of the preceding embodiments, wherein L P1 is-C (R') 2 -substitution, wherein one of the R ' groups is the first R ' group of the four R ' groups, or L P1 Is replaced by-N (R ') -, wherein the R' group is the first R 'group of the four R' groups.
23. The agent of any one of the preceding embodiments, wherein L P1 is-C (R') 2 -substitution, wherein one of the R ' groups is the first R ' group of the four R ' groups.
24. The agent of any one of the preceding embodiments, wherein L P1 Is- [ X ]] p -X 1 -or comprise- [ X ]] p -X 1 -, wherein each X and X 1 Independently an amino acid residue, wherein p is 0 to 10, and X 1 And L is equal to AA1 And (5) bonding.
25. The agent of any one of the preceding embodiments, wherein L P1 is-X 1 -or comprise-X 1 -。
26. The agent of any one of the preceding embodiments, wherein X 1 A first R 'group of the four R' groups.
27. The agent of any one of the preceding embodiments, wherein L AA1 Is an optionally substituted divalent C 2 -C 4 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -，-C(R’)(R AS )-,-Cy-，O-,-S-，-S-S-，-N(R’)-，-C(O)-，-C(s)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-,-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
28. The agent of any one of the preceding embodiments, wherein L AA1 is-N (R ') -C (R') (R) AS )-C(O)-。
29. The agent of any one of the preceding embodiments, wherein L AA1 is-NH-C (R') (R) AS )-C(O)-。
30. The agent of any one of the preceding embodiments, wherein L AS1 Is an optionally substituted divalent C 1 -C 10 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
31. The agent of any one of the preceding embodiments, wherein L AS1 Is an optionally substituted divalent C 1 -C 10 Aliphatic groups wherein one or more methylene units of the group are optionally and independently substituted with-C (R') 2 -Cy-, -O-, -S-, -N (R'), -C (O) -, -S (O) -, or-S (O) 2 -substitution.
32. The agent of any one of the preceding embodiments, wherein L AS1 Is an optionally substituted divalent C 1 -C 10 Aliphatic groups wherein one or more methylene units of the group are optionally and independently replaced by-O-, -S-, or-N (R') -.
33. The agent of any one of the preceding embodiments, wherein L AS1 Is an optionally substituted divalent C 1 -C 10 An alkylene group.
34. The agent of any one of the preceding embodiments, wherein L AS1 Is optionally substituted-CH 2 -。
35. The agent of any one of the preceding embodiments, wherein L AS1 is-CH 2 -。
36. The agent of any one of the preceding embodiments, wherein R AA1 is-CO 2 R。
37. The agent of any one of the preceding embodiments, wherein R AA1 is-CO 2 H。
38. The agent of any one of the preceding embodiments, wherein L AA1 Is an amino acid residue comprising a side chain containing an acidic group.
39. The agent of any one of the preceding embodiments, wherein L AA1 Is X 2 。
40. The agent of any one of the preceding embodiments, wherein L P2 Is a covalent bond or an optionally substituted divalent C 2 -C 6 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
41. The agent of any one of the preceding embodiments, wherein L P2 Is 2 to 10 (2, 3, 4, 5, 6, 7, 8, 9 or 10) atoms in length.
42. The agent of any one of the preceding embodiments, wherein L P2 Is 6 atoms in length.
43. The agent of any one of the preceding embodiments, wherein L P2 Independently replaced by-N (R') -or-C (O) -.
44. The agent of any one of the preceding embodiments, wherein L P2 Independently replaced by-N (R ') -or-C (O) N (R') -.
45. The agent of any one of the preceding embodiments, wherein L P2 Is independently substituted with-N (R '), -C (R') 2 or-C (O) N (R') -substitution.
46. The agent of any one of the preceding embodiments, wherein L P2 Independently replaced by-N (R') -, and L P2 Independently replaced by-C (O) N (R') -.
47. The agent of any one of the preceding embodiments, wherein L P2 Is- [ X ]]pX 4 [X]p' -or comprise- [ X]pX 4 [X]p' -, wherein each X and X 4 Independently are amino acid residues, and p' are each independently 0 to 10.
48. The agent of any one of the preceding embodiments, wherein L P2 Is- [ X ]]pX 3 X 4 [X]p' -or comprise- [ X]pX 3 X 4 [X]p' -, each X, X 3 And X 4 Independently are amino acid residues, and p' are each independently 0 to 10.
49. The agent of any one of the preceding embodiments, wherein L P2 is-X 3 X 4 -or comprise-X 3 X 4 -, wherein X 3 And X 4 Each independently is an amino acid residue, and X 4 And L is equal to AA2 And (5) bonding.
50. The agent of any one of the preceding embodiments, wherein L P2 is-C (R') 2 -alternatively, wherein one of the R ' groups is a second R ' group of the four R ' groups and the other is a third R ' group of the four R ' groups.
51. The agent of any one of the preceding embodiments, wherein X 4 comprising-C (R') 2 -wherein one of the R ' groups is a second R ' group of the four R ' groups and the other is a third R ' group of the four R ' groups.
52. The agent of any of the preceding embodiments, wherein L formed by the first R 'group and the second R' group taken together s Having L as described herein s Radicals and X 1 And X 4 And a bonded structure.
53. The agent of any of the preceding embodiments, wherein L formed by the third R 'group taken together with the fourth R' group s Having L as described herein s Radicals and X 4 And X 11 And a bonded structure.
54. The agent of any one of embodiments 1 to 49, wherein L P2 is-C (R') 2 -substitution, wherein one of the R 'groups is a second R' group.
55. The agent of any one of embodiments 1 to 49, wherein X 3 comprising-C (R') 2 -, wherein one of the R 'groups is a second R' group.
56. The agent of any one of embodiments 1 to 49 and 54 to 55, wherein L P2 is-C (R') 2 -substitution, wherein one of the R 'groups is a third R' group.
57. The agent of any one of embodiments 1 to 49 and 54 to 55, wherein X 4 comprising-C (R') 2 -, wherein one of the R 'groups is a third R' group。
58. The agent of any one of embodiments 1 to 49, wherein L P2 is-C (R') 2 -substitution, wherein one of the R 'groups is a fifth R' group.
59. The agent of any one of embodiments 1 to 49, wherein X 3 comprising-C (R') 2 -, wherein one of the R 'groups is a fifth R' group.
60. The agent of any one of embodiments 1 to 49, wherein L P2 is-C (R') 2 -substitution, wherein one of the R 'groups is a seventh R' group.
61. The agent of any one of embodiments 1 to 49, wherein X 3 comprising-C (R') 2 -, wherein one of the R 'groups is a seventh R' group.
62. The agent of any one of embodiments 1 to 49, wherein L P2 is-C (R') 2 -substitution, wherein one of the R 'groups is a first R' group.
63. The agent of any one of embodiments 1 to 49, wherein X 4 comprising-C (R') 2 -, wherein one of the R 'groups is a first R' group.
64. The agent of any one of the preceding embodiments, wherein L AA2 Is an optionally substituted divalent C 2 -C 4 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -，-C(R’)(R AS )-，-Cy-，-O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
65. The agent of any one of the preceding embodiments, wherein L AA2 is-N (R ') -C (R') (R) AS )-C(O)-。
66. The agent of any one of the preceding embodiments, wherein L AA2 is-NH-C (R') (R) AS )-C(O)-。
67. The agent of any one of the preceding embodiments, wherein L As2 Is an optionally substituted divalent C 1 -C 10 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 N (R') -C (O) S-, or-C (O) O-.
68. The agent of any one of the preceding embodiments, wherein L As2 Is an optionally substituted divalent C 1 -C 10 Aliphatic groups wherein one or more methylene units of the group are optionally and independently substituted with-C (R') 2 -Cy-, -O-, -S-, -N (R'), -C (O) -, -S (O) -, or-S (O) 2 -substitution.
69. The agent of any one of the preceding embodiments, wherein L As2 Is an optionally substituted divalent C 1 -C 10 Aliphatic groups wherein one or more methylene units of the group are optionally and independently replaced by-O-, -S-, or-N (R') -.
70. The agent of any one of the preceding embodiments, wherein L As2 Is an optionally substituted divalent C 1 -C 10 An alkylene group.
71. The agent of any one of the preceding embodiments, wherein L As2 Is optionally substituted-CH 2 -。
72. The agent of any one of the preceding embodiments, wherein L As2 is-CH 2 -。
73. The agent of any one of the preceding embodiments, wherein R AA2 is-CO 2 R。
74. The agent of any one of the preceding embodiments, wherein R AA2 is-CO 2 H。
75. The agent of any one of the preceding embodiments, wherein L AA2 Is an amino acid residue comprising a side chain containing an acidic group.
76. The agent of any one of the preceding embodiments, wherein L AA2 Is X 5 。
77. The agent of any one of the preceding embodiments, wherein L P3 Is 0 to 10 (0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) atoms in length.
78. The agent of any one of the preceding embodiments, wherein L P3 Is a covalent bond or an optionally substituted divalent C 2 -C 6 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
79. The agent of any one of the preceding embodiments, wherein L P3 Is 2 to 10 (2, 3, 4, 5, 6, 7, 8, 9 or 10) atoms in length.
80. The agent of any one of the preceding embodiments, wherein L P3 Is 6 atoms in length.
81. The agent of any one of the preceding embodiments, wherein L P3 Independently replaced by-N (R') -or-C (O) -.
82. The agent of any one of the preceding embodiments, wherein L P3 Independently replaced by-N (R ') -or-C (O) N (R') -.
83. The agent of any one of the preceding embodiments, wherein L P3 Is independently substituted with-N (R '), -C (R') 2 -or-C (O) N (R') -substitution.
84. The agent of any one of the preceding embodiments, wherein L P3 Independently replaced by-N (R') -, and L P3 Independently replaced by-C (O) N (R') -.
85. The foregoing embodimentThe agent of any one of the schemes, wherein L P3 Is- [ X ]]pX 6 X 7 [X]p' -or comprise- [ X]pX 6 X 7 [X]p' -, X, X 6 And X 7 Each independently is an amino acid residue, and p' are each independently 0 to 10.
86. The agent of any one of the preceding embodiments, wherein L P3 is-X 6 X 7 -or comprise-X 6 X 7 -, wherein X 6 And X 7 Each independently is an amino acid residue, and X 7 And L is equal to AA3 And (5) bonding.
87. The agent of any one of the preceding embodiments, wherein L P3 is-C (R') 2 -alternatively, wherein one of the R ' groups is a fifth R ' group, a sixth R ' group, a seventh R ' group or an eighth R ' group.
88. The agent of any one of the preceding embodiments, wherein X 7 comprising-C (R') 2 -, wherein one of the R ' groups is a fifth R ' group, a sixth R ' group, a seventh R ' group, or an eighth R ' group.
89. The agent of any of embodiments 87-88, wherein the R 'group is a fifth R' group.
90. The agent of any of embodiments 87-88, wherein the R 'group is a sixth R' group.
91. The agent of any one of embodiments 87-88, wherein the R 'group is a seventh R' group.
92. The agent of any of embodiments 87-88, wherein the R 'group is an eighth R' group.
93. The agent of any one of the preceding embodiments, wherein L P3 Is a covalent bond.
94. The agent of any one of the preceding embodiments, wherein L AA3 Is an optionally substituted divalent C 2 -C 4 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -，-C(R’)(R AS )-，-Cy-，-O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
95. The agent of any one of the preceding embodiments, wherein L AA3 is-N (R ') -C (R') (R) AS )-C(O)-。
96. The agent of any one of the preceding embodiments, wherein L AA3 is-NH-C (R') (R) AS )-C(O)-。
97. The agent of any one of the preceding embodiments, L AS3 Is an optionally substituted divalent C 1 -C 10 Aliphatic groups wherein one or more methylene units of the group are optionally and independently substituted with-C (R') 2 -，-Cy-,-O-，-S-，-S-S-,-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 N (R') -C (O) S-, or-C (O) O-.
98. The agent of any one of the preceding embodiments, wherein R AS is-L AS3 -R AA3 Wherein L is AS3 Is an optionally substituted divalent C 1 -C 10 Aliphatic groups wherein one or more methylene units of the group are optionally and independently substituted with-C (R') 2 -Cy-, -O-, -S-, -N (R'), -C (O) -, -S (O) -, or-S (O) 2 -substitution.
99. The agent of any one of the preceding embodiments, wherein L AS3 Is an optionally substituted divalent C 1 -C 10 Aliphatic groups wherein one or more methylene units of the group are optionally and independently replaced by-O-, -S-, or-N (R') -.
100. The agent of any one of the preceding embodiments, wherein L AS3 Is an optionally substituted divalent C 1 -C 10 An alkylene group.
101. The agent of any one of the preceding embodiments, wherein L AS3 Is optionally substituted-CH 2 -。
102. The agent of any one of the preceding embodiments, wherein L AS3 is-CH 2 -。
103. The agent of any one of the preceding embodiments, wherein R AA3 is-CO 2 R。
104. The agent of any one of the preceding embodiments, wherein R AA3 is-CO 2 H。
105. The agent of any one of the preceding embodiments, wherein L AA3 Is an amino acid residue comprising a side chain containing an acidic group.
106. The agent of any one of the preceding embodiments, wherein L AA3 Is X 6 。
107. The agent of any one of embodiments 1 to 102, wherein L AA3 Are amino acid residues comprising hydrophobic side chains.
108. The agent of any one of embodiments 1 to 102, wherein R AA3 Is a hydrophobic group.
109. The agent of any one of embodiments 1 to 102, wherein R AA3 Is optionally substituted C 1-6 An aliphatic group.
110. The agent of any one of embodiments 1 to 102, wherein R AA3 Is C 1-6 An aliphatic group.
111. The agent of any one of embodiments 1 to 102, wherein R AA3 Is C 1-6 An alkyl group.
112. The agent of any one of embodiments 1 to 102, wherein L AA3 Is X 8 。
113. The agent of any one of the preceding embodiments, wherein L P4 Is a covalent bond or an optionally substituted divalent C 2 -C 6 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -,-Cy-,-O-，-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
114. The agent of any one of the preceding embodiments, wherein L P4 Is 0 to 10 (0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) atoms in length.
115. The agent of any one of the preceding embodiments, wherein L P4 Is 2 to 10 (2, 3, 4, 5, 6, 7, 8, 9 or 10) atoms in length.
116. The agent of any one of the preceding embodiments, wherein L P4 Is 6 atoms in length.
117. The agent of any one of the preceding embodiments, wherein L P4 Independently replaced by-N (R') -or-C (O) -.
118. The agent of any one of the preceding embodiments, wherein L P4 Independently replaced by-N (R ') -or-C (O) N (R') -.
119. The agent of any one of the preceding embodiments, wherein L P4 Is independently substituted with-N (R '), -C (R') 2 or-C (O) N (R') -substitution.
120. The agent of any one of the preceding embodiments, wherein L P4 Independently replaced by-N (R') -, and L P4 Independently replaced by-C (O) N (R') -.
121. The agent of any one of the preceding embodiments, wherein L P4 Is- [ X ]]pX 7 X 8 [X]p' -or comprise- [ X]pX 7 X 8 [X]p' -, X, X 7 And X 8 Each independently is an amino acid residue, and p' are each independently 0 to 10.
122. The agent of any one of the preceding embodiments, wherein L P4 is-X 7 X 8 -or comprise-X 7 X 8 -, wherein X 7 And X 8 Each independently is an amino acid residue, and X 8 And L is equal to AA4 And (5) bonding.
123. The foregoing embodimentThe agent of any one of the schemes, wherein L P4 is-C (R') 2 -alternatively, wherein one of the R ' groups is a fifth R ' group, a sixth R ' group, a seventh R ' group or an eighth R ' group.
124. The agent of any one of the preceding embodiments, wherein X 7 comprising-C (R') 2 -, wherein one of the R ' groups is a fifth R ' group, a sixth R ' group, a seventh R ' group, or an eighth R ' group.
125. The agent of any one of embodiments 123 to 124, wherein the R 'group is a fifth R' group.
126. The agent of any one of embodiments 123 to 124, wherein the R 'group is a sixth R' group.
127. The agent of any one of embodiments 123 to 124, wherein the R 'group is a seventh R' group.
128. The agent of any one of embodiments 123-124, wherein the R 'group is an eighth R' group
129. The agent of any one of embodiments 1 to 112, wherein L P4 Is a covalent bond.
130. The agent of any one of the preceding embodiments, wherein L AA4 Is an optionally substituted divalent C 2 -C 4 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -，-C(R’)(R AS )-,-Cy-，-O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
131. The agent of any one of the preceding embodiments, wherein L AA4 is-N (R ') -C (R') (R) AS )-C(O)-。
132. The agent of any one of the preceding embodiments, wherein L AA4 is-NH-C (R') (R) AS )-C(O)-。
133. Any of the foregoing embodimentsThe medicament, wherein L AS4 Is an optionally substituted divalent C 1 -C 10 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
134. The agent of any one of the preceding embodiments, wherein L AS4 Is an optionally substituted divalent C 1 -C 10 Aliphatic groups wherein one or more methylene units of the group are optionally and independently substituted with-C (R') 2 -Cy-, -O-, -S-, -N (R'), -C (O) -, -S (O) -, or-S (O) 2 -substitution.
135. The agent of any one of the preceding embodiments, L AS4 Is an optionally substituted divalent C 1 -C 10 Aliphatic groups wherein one or more methylene units of the group are optionally and independently replaced by-O-, -S-, or-N (R') -.
136. The agent of any one of the preceding embodiments, wherein L AS4 Is an optionally substituted divalent C 1 -C 10 An alkylene group.
137. The agent of any one of the preceding embodiments, wherein L AS4 Is optionally substituted-CH 2 -。
138. The agent of any one of the preceding embodiments, wherein L AS4 is-CH 2 -。
139. The agent of any one of the preceding embodiments, wherein R AA4 Is an optionally substituted 6 to 14 membered aryl.
140. The agent of any one of the preceding embodiments, wherein R AA4 Is an optionally substituted phenyl group.
141. The agent of any one of the preceding embodiments, wherein R AA4 Is phenyl.
142. Embodiment 1 to138, wherein R AA4 Is an optionally substituted 5-to 14-membered heteroaryl having 1 to 6 heteroatoms.
143. The agent of any one of embodiments 1 to 138, wherein R AA4 Is an optionally substituted 5 membered monocyclic heteroaryl having 1 to 4 heteroatoms.
144. The agent of any one of embodiments 1 to 138, wherein R AA4 Optionally substituted
145. The agent of any one of embodiments 1 to 138, wherein R AA4 Is an optionally substituted 9 membered bicyclic heteroaryl having 1 to 4 heteroatoms.
146. The agent of any one of embodiments 1 to 138, wherein R AA4 Is an optionally substituted 10 membered bicyclic heteroaryl having 1 to 4 heteroatoms.
147. The agent of any one of embodiments 1 to 138, wherein R AA4 Is optionally substituted
148. The agent of any one of embodiments 1 to 138, wherein R AA4 Is optionally substituted
149. The agent of any one of the preceding embodiments, wherein L AA4 Is an amino acid residue.
150. The agent of any one of the preceding embodiments, wherein L AA4 Is X 9 。
151. The agent of any one of the preceding embodiments, wherein L P5 Is a covalent bond or an optionally substituted divalent C 2 -C 6 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
152. The agent of any one of the preceding embodiments, wherein L P5 Is 2 to 10 (2, 3, 4, 5, 6, 7, 8, 9 or 10) atoms in length.
153. The agent of any one of the preceding embodiments, wherein L P5 Is 6 atoms in length.
154. The agent of any one of the preceding embodiments, wherein L P5 Independently replaced by-N (R') -or-C (O) -.
155. The agent of any one of the preceding embodiments, wherein L P5 Independently replaced by-N (R ') -or-C (O) N (R') -.
156. The agent of any one of the preceding embodiments, wherein L P5 Is independently substituted with-N (R '), -C (R') 2 or-C (O) N (R') -substitution.
157. The agent of any one of the preceding embodiments, wherein L P5 Independently replaced by-N (R') -, and L P5 Independently replaced by-C (O) N (R') -.
158. The agent of any one of the preceding embodiments, wherein L P5 is-C (R') 2 -substitution, wherein one of the R ' groups is a second R ' group or a fourth R ' group.
159. The agent of any one of the preceding embodiments, wherein L P5 Is- [ X ]]pX 10 X 11 [X]p' -or comprise- [ X]pX 10 X 11 [X]p' -, X, X 10 And X 11 Each independently is an amino acid residue, and p' are each independently 0 to 10.
160. The agent of any one of the preceding embodiments, wherein L P5 is-X 10 X 11 -or comprise-X 10 X 11 -, wherein X 10 And X 11 Each independently is an amino acid residue, and X 11 And L is equal to AA5 And (5) bonding.
161. The agent of any one of the preceding embodiments, wherein X 11 comprising-C (R') 2 -, wherein one of the R ' groups is a second R ' group or a fourth R ' group.
162. The agent of embodiment 158 or 161 wherein one of the R 'groups is a second R' group.
163. The agent of embodiment 158 or 161 wherein one of the R 'groups is a fourth R' group.
164. The agent of any one of the preceding embodiments, wherein L P5 is-C (R') 2 -alternatively, wherein one of the R ' groups is a fifth R ' group, a sixth R ' group, a seventh R ' group or an eighth R ' group.
165. The agent of any one of the preceding embodiments, wherein X 10 comprising-C (R') 2 -, wherein one of the R ' groups is a fifth R ' group, a sixth R ' group, a seventh R ' group, or an eighth R ' group.
166. The agent of any one of embodiments 164 to 165 wherein the R 'group is a fifth R' group.
167. The agent of any one of embodiments 164 to 165 wherein the R 'group is a sixth R' group.
168. The agent of any one of embodiments 164 to 165 wherein the R 'group is a seventh R' group.
169. The agent of any one of embodiments 164 to 165 wherein the R 'group is an eighth R' group.
170. The agent of any one of the preceding embodiments, wherein L AA5 Is an optionally substituted divalent C 2 -C 4 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -，-C(R’)(R AS )-，-Cy-，-O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
171. The agent of any one of the preceding embodiments, wherein L AA5 is-N (R ') -C (R') (R) AS )-C(O)-。
172. The agent of any one of the preceding embodiments, wherein L AA5 is-NH-C (R') (R) AS )-C(O)-。
173. The agent of any one of the preceding embodiments, wherein L AS5 Is an optionally substituted divalent C 1 -C 10 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) ２ n (R') -C (O) S-, or-C (O) O-.
174. The agent of any one of the preceding embodiments, wherein L AS5 Is an optionally substituted divalent C 1 -C 10 Aliphatic groups wherein one or more methylene units of the group are optionally and independently substituted with-C (R') 2 -Cy-, -O-, -S-, -N (R'), -C (O) -, -S (O) -, or-S (O) 2 -substitution.
175. The agent of any one of the preceding embodiments, wherein L AS5 Is an optionally substituted divalent C 1 -C 10 Aliphatic groups wherein one or more methylene units of the group are optionally and independently replaced by-O-, -S-, or-N (R') -.
176. The agent of any one of the preceding embodiments, wherein L AS5 Is an optionally substituted divalent C 1 -C 10 An alkylene group.
177. The agent of any one of the preceding embodiments, wherein L AS5 Is optionally substituted-CH 2 -。
178. The agent of any one of the preceding embodiments, wherein L AS5 is-CH 2 -。
179. The agent of any one of the preceding embodiments, wherein R AA5 Is an optionally substituted 6 to 14 membered aryl.
180. The agent of any one of the preceding embodiments, wherein R AA5 Is an optionally substituted phenyl group.
181. The agent of any one of the preceding embodiments, wherein R AA5 Is phenyl.
182. The agent of any one of embodiments 1 to 169, wherein R AA5 Is an optionally substituted 5-to 14-membered heteroaryl having 1 to 6 heteroatoms.
183. The agent of any one of embodiments 1 to 169, wherein R AA5 Is an optionally substituted 5 membered monocyclic heteroaryl having 1 to 4 heteroatoms.
184. The agent of any one of embodiments 1 to 169, wherein R AA5 Is optionally substituted
185. The agent of any one of embodiments 1 to 169, wherein R AA5 Is an optionally substituted 9 membered bicyclic heteroaryl having 1 to 4 heteroatoms.
186. The agent of any one of embodiments 1 to 169, wherein R AA5 Is an optionally substituted 10 membered bicyclic heteroaryl having 1 to 4 heteroatoms.
187. The agent of any one of embodiments 1 to 169, wherein R AA5 Is optionally substituted
188. The agent of any one of embodiments 1 to 169, wherein R AA5 Is optionally substituted
189. The agent of any one of the preceding embodiments, wherein L AA5 Is an amino acid residue.
190. The agent of any one of the preceding embodiments, wherein L AA5 Is X 12 。
191. The agent of any one of the preceding embodiments, wherein L P6 Is a covalent bond or an optionally substituted divalent C 2 -C 6 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -，-Cy-,-O-，-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)- , -S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
192. The agent of any one of the preceding embodiments, wherein L P6 Is 0 to 10 (0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) atoms in length.
193. The agent of any one of the preceding embodiments, wherein L P6 Is a covalent bond.
194. The agent of any one of the preceding embodiments, wherein L AA6 Is an optionally substituted divalent C 2 -C 4 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -，-C(R’)(R AS )-，-Cy-，-O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-,-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
195. The agent of any one of the preceding embodiments, wherein L AA6 The methylene units of which are-C (R') (R AS ) -substitution, wherein R AS is-L AS -R AA6 Wherein L is AS Is an optionally substituted divalent C 1 -C 10 Aliphatic groups, wherein one or more of the groupsThe individual methylene units are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
196. The agent of any one of the preceding embodiments, wherein L AA6 The methylene units of which are-C (R') (R AS ) -substitution, wherein R AS is-L AS -R AA6 Wherein L is AS Is an optionally substituted divalent C 1 -C 10 Aliphatic groups wherein one or more methylene units of the group are optionally and independently substituted with-C (R') 2 -Cy-, -O-, -S-, -N (R'), -C (O) -, -S (O) -, or-S (O) 2 -substitution.
197, wherein L is AA6 The methylene units of which are-C (R') (R AS ) -substitution, wherein R AS is-L AS -R AA6 Wherein L is AS Is an optionally substituted divalent C 1 -C 10 Aliphatic groups wherein one or more methylene units of the group are optionally and independently replaced by-O-, -S-, or-N (R') -.
198. The agent of any one of the preceding embodiments, wherein L AA6 The methylene units of which are-C (R') (R AS ) -substitution, wherein R AS is-L AS -R AA6 Wherein L is AS Is an optionally substituted divalent C 1 -C 10 An alkylene group.
199. The agent of any one of the preceding embodiments, wherein L AA6 The methylene units of which are-C (R') (R AS ) -substitution, wherein R AS is-CH 2 -R AA6 。
200. The agent of any one of the preceding embodiments, wherein R AA6 Is an optionally substituted 6 to 14 membered aryl.
201. The agent of any one of the preceding embodiments, wherein R AA6 Is optionally substitutedPhenyl groups of (a).
202. The agent of any one of the preceding embodiments, wherein R AA6 Is phenyl.
203. The agent of any one of embodiments 1 to 193, wherein R AA6 Is an optionally substituted 5-to 14-membered heteroaryl having 1 to 6 heteroatoms.
204. The agent of any one of embodiments 1 to 193, wherein R AA6 Is an optionally substituted 5 membered monocyclic heteroaryl having 1 to 4 heteroatoms.
205. The agent of any one of embodiments 1 to 193, wherein R AA6 Is optionally substituted
206. The agent of any one of embodiments 1 to 193, wherein R AA6 Is an optionally substituted 9 membered bicyclic heteroaryl having 1 to 4 heteroatoms.
207. The agent of any one of embodiments 1 to 193, wherein R AA6 Is an optionally substituted 10 membered bicyclic heteroaryl having 1 to 4 heteroatoms.
208. The agent of any one of embodiments 1 to 193, wherein R AA6 Is optionally substituted
209. The agent of any one of embodiments 1 to 193, wherein R AA6 Is optionally substituted
210. The agent of any one of the preceding embodiments, wherein L AA6 Is an amino acid residue.
211. The agent of any one of the preceding embodiments, wherein L AA6 Is X 13 。
212. The agent of any one of the preceding embodiments, wherein L P7 Is a covalent bond or anyOptionally substituted divalent C 1 -C 10 Aliphatic or heteroaliphatic groups having from 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -,-Cy-,-O-,-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
213. The agent of any one of the preceding embodiments, wherein L P7 Is 0 to 20 (e.g., 0 to 15, 0 to 10, 0 to 5, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) atoms in length.
214. The agent of any one of the preceding embodiments, wherein L P7 is-X 14 -[X]p' -or containing-X 14 -[X]p '-, where p' is 0 to 10, X and X 14 Each independently is an amino acid residue, and X 14 And L is equal to AA6 And (5) bonding.
215. The agent of any one of the preceding embodiments, wherein L P7 is-C (R') 2 -substitution, wherein one of the R ' groups is a sixth R ' group or an eighth R ' group.
216. The agent of any one of the preceding embodiments, wherein X 14 comprising-C (R') 2 -, wherein one of the R ' groups is a fifth R ' group, a sixth R ' group, a seventh R ' group, or an eighth R ' group.
217. The agent of any one of embodiments 215-216, wherein the R 'group is a sixth R' group.
218. The agent of any one of embodiments 215-216, wherein the R 'group is an eighth R' group.
219. The agent of any one of the preceding embodiments, wherein L RN Is a covalent bond, or an optionally substituted divalent C 1 -C 10 Aliphatic or heteroaliphatic groups having 1 to 10 heteroatoms, wherein one or more methylene units of the groupThe elements are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
220. The agent of any one of the preceding embodiments, wherein L RN Is 0 to 20 (0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20) atoms in length.
221. The agent of any one of the preceding embodiments, wherein R N Is R' -L RN -, wherein R' is-C (O) R, -CO 2 R or-SO 2 R。
222. The agent of any one of the preceding embodiments, wherein R N Is R ', wherein R' is-C (O) R, -CO 2 R or-SO 2 R。
223. The agent of any one of the preceding embodiments, wherein L RC Is a covalent bond or an optionally substituted divalent C 1 -C 10 Aliphatic or heteroaliphatic groups having from 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -,-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
224. The agent of any one of the preceding embodiments, wherein L RC Is 0 to 20 (0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20) atoms in length.
225. The agent of any one of the preceding embodiments, wherein R C is-O-L RC -R 'or-N (R') -L RC -R’。
226. The agent of any one of the preceding embodiments, wherein R C is-OR 'OR-N (R') 2 WhereinEach R' is independently R.
227. A medicament comprising one or more of:
a first acidic group (e.g., belonging to a first acidic amino acid residue);
a second acidic group (e.g., belonging to a second acidic amino acid residue);
a first aromatic group (e.g., belonging to a first aromatic amino acid residue);
a second aromatic group (e.g., belonging to the first aromatic amino acid residue); and
A third aromatic group (e.g., belonging to a third aromatic amino acid residue).
228. A medicament, comprising:
a first acidic group (e.g., belonging to a first acidic amino acid residue);
a second acidic group (e.g., belonging to a second acidic amino acid residue);
a first aromatic group (e.g., belonging to a first aromatic amino acid residue);
a second aromatic group (e.g., belonging to the first aromatic amino acid residue); and
a third aromatic group (e.g., belonging to a third aromatic amino acid residue).
229. A medicament, comprising:
a first acidic group (e.g., belonging to a first acidic amino acid residue);
a second acidic group (e.g., belonging to a second acidic amino acid residue);
a third acidic group (e.g., belonging to a third acidic amino acid residue);
a first aromatic group (e.g., belonging to a first aromatic amino acid residue);
a second aromatic group (e.g., belonging to the first aromatic amino acid residue); and
a third aromatic group (e.g., belonging to a third aromatic amino acid residue).
230. A medicament, comprising:
a first acidic group (e.g., belonging to a first acidic amino acid residue);
a second acidic group (e.g., belonging to a second acidic amino acid residue);
Hydrophobic groups (e.g., belonging to hydrophobic amino acid residues);
a first aromatic group (e.g., belonging to a first aromatic amino acid residue);
a second aromatic group (e.g., belonging to the first aromatic amino acid residue); and
a third aromatic group (e.g., belonging to a third aromatic amino acid residue).
231. A medicament, comprising:
a first acidic group (e.g., belonging to a first acidic amino acid residue);
a second acidic group (e.g., belonging to a second acidic amino acid residue);
a third acidic group (e.g., belonging to a third acidic amino acid residue);
hydrophobic groups (e.g., belonging to hydrophobic amino acid residues);
a first aromatic group (e.g., belonging to a first aromatic amino acid residue);
a second aromatic group (e.g., belonging to the first aromatic amino acid residue); and
a third aromatic group (e.g., belonging to a third aromatic amino acid residue).
232. The agent of any of embodiments 227 to 231 wherein the first acidic group belongs to a first acidic amino acid residue.
233. The agent of any one of embodiments 227 to 231 wherein the first acidic group belongs to L of any one of the preceding embodiments AA1 。
234. The agent of any of embodiments 227 to 231 wherein the first acidic group belongs to a first acidic amino acid residue, which is X 2 。
235. The agent of any of embodiments 227-234 wherein the second acidic group belongs to a second acidic amino acid residue.
236. The agent of any one of embodiments 227 to 234 wherein the second acidic group belongs to L of any one of the preceding embodiments AA2 。
237. The agent of any of embodiments 227 to 234 wherein the second acidic group belongs to a second acidic amino acid residue, which is X 5 。
238. The agent of any of embodiments 227 to 237 wherein the third acidic group belongs to a third acidic amino acid residue.
239. The agent of any one of embodiments 227 to 237 wherein the third acidic group belongs to L of any one of the previous embodiments AA3 Wherein L is AA3 Comprising an acidic group.
240. The agent of any of embodiments 227 to 237 wherein the third acidic group belongs to a third acidic amino acid residue, which is X 6 。
241. The agent of any of embodiments 227 to 240 wherein the hydrophobic group belongs to a hydrophobic acidic amino acid residue.
242. The agent of any of embodiments 227 to 240 wherein the hydrophobic group belongs to L of any of the preceding embodiments AA3 Wherein L is AA3 Comprising hydrophobic groups.
243. The agent of any of embodiments 227 to 240 wherein the hydrophobic group is a hydrophobic acidic amino acid residue, which is X 8 。
244. The agent of any of embodiments 227 to 243 wherein the first aromatic group belongs to a first aromatic amino aromatic residue.
245. The agent of any one of embodiments 227 to 243 wherein the first aromatic group belongs to L of any one of the preceding embodiments AA4 。
246. The medicament of any of embodiments 227 to 243, wherein the first aromatic group is a first aromatic amino aromatic residueThe radical being X 9 。
247. The agent of any of embodiments 227-246 wherein the second aromatic group belongs to a second aromatic amino aromatic residue.
248. The agent of any one of embodiments 227 to 246 wherein the second aromatic group belongs to L of any one of the preceding embodiments AA5 。
249. The medicament of any of embodiments 227 to 246, wherein the second aromatic group is a second aromatic amino aromatic residue, which is X 12 。
250. The agent of any of embodiments 227-249 wherein the third aromatic group is a third aromatic amino aromatic residue.
251. The agent of any of embodiments 227 through 249 wherein the third aromatic group is L as in any of the previous embodiments AA3 Wherein L is AA6 Comprising aromatic groups.
252. The medicament of any of embodiments 227 to 249, wherein the third aromatic group is a third aromatic amino aromatic residue, which is X 13 。
253. The agent of any one of the preceding embodiments, wherein the distance between the first acidic group and the second acidic group is about the distance between the acidic groups of two acidic amino acid residues of the peptide motif, wherein there are two amino acid residues between the two acidic amino acid residues.
254. The agent of any one of the preceding embodiments, wherein the first acidic amino acid residue is at position N and the second acidic amino acid residue is at position n+3.
255. The agent of any one of the preceding embodiments, wherein the distance between the first acidic group and the third acidic group is about the distance between the acidic groups of two acidic amino acid residues of the peptide motif, wherein there are three amino acid residues between the two acidic amino acid residues.
256. The agent of any one of the preceding embodiments, wherein the first acidic amino acid residue is at position N and the third acidic amino acid residue is at position n+4.
257. The agent of any one of the preceding embodiments, wherein the distance between the first acidic group and the hydrophobic group is about the distance between the acidic group of the acidic amino acid residue of the peptide motif and the hydrophobic group of the hydrophobic amino acid residue, wherein there are five amino acid residues between the first acidic amino acid residue and the hydrophobic amino acid residue.
258. The agent of any one of the preceding embodiments, wherein the first acidic amino acid residue is at position N and the hydrophobic amino acid residue is at position n+6.
259. The agent of any one of the preceding embodiments, wherein the distance between the first acidic group and the first aromatic group is about the distance between the acidic group of the first acidic amino acid residue and the aromatic group of the aromatic amino acid residue of the peptide motif, wherein there are six amino acid residues between the first acidic amino acid residue and the first aromatic amino acid residue.
260. The agent of any one of the preceding embodiments, wherein the first acidic amino acid residue is at position N and the first aromatic amino acid residue is at position n+7.
261. The agent of any one of the preceding embodiments, wherein the distance between the first aromatic group and the second aromatic group is about the distance between the aromatic groups of two aromatic amino acid residues of the peptide motif, wherein there are two amino acid residues between the two aromatic amino acid residues.
262. The agent of any one of the preceding embodiments, wherein the first aromatic amino acid residue is at position M and the second aromatic amino acid residue is at position m+3.
263. The agent of any one of the preceding embodiments, wherein the distance between the first aromatic group and the third aromatic group is about the distance between the aromatic groups of two aromatic amino acid residues of the peptide motif, wherein there are three amino acid residues between the two aromatic amino acid residues.
264. The agent of any one of the preceding embodiments, wherein the first aromatic amino acid residue is at position N and the third aromatic amino acid residue is at position m+4).
265. The agent of any one of the preceding embodiments, wherein N is 1 to 7.
266. The agent of any one of the preceding embodiments, wherein N is 1, 2, 3, 4, or 5.
267. The agent of any one of the preceding embodiments, wherein N is 1.
268. The agent of any one of the preceding embodiments, wherein N is 2.
269. The agent of any one of the preceding embodiments, wherein N is 3.
270. The agent of any one of the preceding embodiments, wherein N is 4.
271. The agent of any one of the preceding embodiments, wherein N is 5.
272. The agent of any one of the preceding embodiments, wherein M is n+7.
273. The agent of any one of the preceding embodiments, wherein M is 8 to 16.
274. The agent of any one of the preceding embodiments, wherein M is 8.
275. The agent of any one of the preceding embodiments, wherein M is 9.
276. The agent of any one of the preceding embodiments, wherein M is 10.
277. The agent of any one of the preceding embodiments, wherein M is 11.
278. The agent of any one of the preceding embodiments, wherein M is 12.
279. The agent of any one of the preceding embodiments, wherein M is 13.
280. The agent of any one of embodiments 253-279, wherein the peptide motif is an alpha-helical motif, wherein each amino acid residue is independently an alpha amino acid residue.
281. The agent of embodiment 280, wherein the peptide motif is stapled.
282. The agent of embodiment 281 wherein there are two staple-like structures in the peptide motif.
283. The agent of embodiment 281 wherein there are three staple-like structures in the peptide motif.
284. The agent of embodiment 281 wherein the peptide motif has four staple-like structures therein.
285. The agent of any one of embodiments 253-279, wherein the peptide motif is or comprises an agent described in the tables herein (e.g., I-xxxx, wherein xxxx is a number (e.g., I-1, I-10, I-100, I-1000, etc.)).
286. The agent of any one of the preceding embodiments, wherein the first acidic group interacts with Lys312 or with the corresponding amino acid residue thereof when the agent is contacted with a β -catenin polypeptide.
287. The agent of any one of the preceding embodiments, wherein when the agent is contacted with a β -catenin polypeptide, the first acidic group interacts with Gly307 or with the corresponding amino acid residue.
288. The agent of any one of the preceding embodiments, wherein the second acidic group interacts with Asn387 or with a corresponding amino acid residue thereof when the agent is contacted with a β -catenin polypeptide.
289. The agent of any one of the preceding embodiments, wherein the second acidic group interacts with Trp383 or with the corresponding amino acid residue thereof when the agent is contacted with the β -catenin polypeptide.
290. The agent of any one of the preceding embodiments, wherein the third acidic group interacts with Tyr306 or with the corresponding amino acid residue thereof when the agent is contacted with a β -catenin polypeptide.
291. The agent of any one of the preceding embodiments, wherein the hydrophobic group interacts with Trp383 or with the corresponding amino acid residue thereof when the agent is contacted with a β -catenin polypeptide.
292. The agent of any one of the preceding embodiments, wherein the first aromatic group interacts with Lys345 or with the corresponding amino acid residue thereof when the agent is contacted with a β -catenin polypeptide.
293. The agent of any one of the preceding embodiments, wherein the first aromatic group interacts with Trp383 or with the corresponding amino acid residue thereof when the agent is contacted with a β -catenin polypeptide.
294. The agent of any one of the preceding embodiments, wherein the second aromatic group interacts with Trp383 or with the corresponding amino acid residue thereof when the agent is contacted with the β -catenin polypeptide.
295. The agent of any one of the preceding embodiments, wherein the second aromatic group interacts with Asn415 or with the corresponding amino acid residue thereof when the agent is contacted with a β -catenin polypeptide.
296. The agent of any one of the preceding embodiments, wherein the third aromatic group interacts with Gln379 or with the corresponding amino acid residue thereof when the agent is contacted with a β -catenin polypeptide.
297. The agent of any one of the preceding embodiments, wherein the third aromatic group interacts with Leu382 or with the corresponding amino acid residue when the agent is contacted with a β -catenin polypeptide.
298. The agent of any one of the preceding embodiments, wherein the third aromatic group interacts with Val416 or with the corresponding amino acid residue when the agent is contacted with a β -catenin polypeptide.
299. The agent of any one of the preceding embodiments, wherein the third aromatic group interacts with Asn415 or with the corresponding amino acid residue thereof when the agent is contacted with a β -catenin polypeptide.
300. The agent of any one of the preceding embodiments, wherein the third aromatic group interacts with Trp383 or with the corresponding amino acid residue thereof when the agent is contacted with the β -catenin polypeptide.
301. The agent of any one of the preceding embodiments, wherein the agent is or comprises a peptide.
302. The agent of any one of the preceding embodiments, wherein the agent is a peptide.
303. The agent of any one of the preceding embodiments, wherein the peptide is a stapled peptide comprising two or more staple-like structures.
304. The agent of any one of the preceding embodiments, wherein the peptide is a stapled peptide comprising three or more staple-like structures.
305. The agent of any one of the preceding embodiments, wherein the peptide is a stapled peptide comprising three and no more than three staple-like structures.
306. The agent of any one of the preceding embodiments, wherein the peptide is a stapled peptide comprising four and no more than four staple-like structures.
307. The agent of any one of the preceding embodiments, wherein the first acidic group, the second acidic group, the third acidic group, the hydrophobic group, the first aromatic group, the second aromatic group, and the third aromatic group (if present) are present in the N-to-C direction of the peptide.
308. The agent of any one of the preceding embodiments, wherein the agent is or comprises a helical structure.
309. A medicament, comprising:
X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 ，
wherein:
X 1 ，X 2 ，X 3 ，X 4 ，X 5 ，X 6 ，X 7 ，X 8 ，X 9 ，X 10 ，X 11 ，X 12 ，X 13 and X 14 Each independently is an amino acid residue, wherein:
X 2 comprising a side chain containing an acidic or polar group;
X 5 comprising a side chain containing an acidic or polar group; and is also provided with
X 9 、X 12 And X 13 Each comprising a side chain containing an optionally substituted aromatic group.
310. An agent, wherein the agent is or comprises a peptide comprising:
[X 0 ] p0 X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 ，
wherein:
p0, p15, p16 and p17 are each independently 0 or 1;
X 0 ，X 1 ,X 2 ，X 3 ，X 4 ，X 5 ，X 6 ，X 7 ，X 8 ，X 9 ，X 10 ，X 11 ，X 12 ，X 13 ，X 14 ，X 15 ，X 16 and X 17 Each independently is an amino acid residue, wherein:
X 2 comprising a side chain containing an acidic or polar group;
X 5 comprising a side chain containing an acidic or polar group;
X 9 、X 12 and X 13 Each comprising a side chain containing an optionally substituted aromatic group.
311. A medicament, comprising:
X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 ，
wherein:
X 1 ，X 2 ，X 3 ，X 4 ，X 5 ，X 6 ，X 7 ，X 8 ，X 9 ，X 10 ，X 11 ，X 12 ，X 13 and X 14 Each independently of the otherIs an amino acid residue, wherein:
X 2 comprising a side chain containing an acidic or polar group;
X 5 comprising a side chain containing an acidic or polar group;
X 13 comprising a side chain comprising an optionally substituted aromatic group; and is also provided with
X 1 、X 3 、X 4 、X 7 、X 10 、X 11 And X 14 Each independently is an amino acid residue suitable for stapling, or each independently is stapling.
312. An agent, wherein the agent is or comprises:
X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 [X 14 ] p14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 [X 18 ] p18 [X 19 ] p19 [X 20 ] p20 [X 21 ] p21 [X 22 ] p22 [X 23 ] p23 ，
Wherein p14, p15, p16, p17, p18, p19, p20, p21, p22 and p23 are each independently 0 or 1, and X 1 ，X 2 ，X 3 ，X 4 ，X 5 ，X 6 ，X 7 ，X 8 ，X 9 ，X 10 ，X 11 ，X 12 ，X 13 ，X 14 ，X 15 ，X 16 ，X 17 ，X 18 ，X 19 ，X 20 ，X 21 ，X 22 And X 23 Each independently is an amino acid residue.
313. An agent, wherein the agent is or comprises:
[X] p X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 [X] p’ ，
wherein:
p15, p16 and p17 are each independently 0 or 1;
p and p' are each independently 0 to 10;
X，X 1 ，X 2 ，X 3 ，X 4 ，X 5 ，X 6 ，X 7 ，X 8 ，X 9 ，X 10 ，X 11 ，X 12 ，X 13 ，X 14 ，X 15 ，X 16 and X 17 Each independently is an amino acid residue.
314. An agent, wherein the agent is or comprises a peptide comprising:
[X 0 ] p0 X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 ，
wherein:
p0, p15, p16 and p17 are each independently 0 or 1;
X 0 ,X 1 ,X 2 ,X 3 ,X 4 ,X 5 ,X 6 ,X 7 ,X 8 ,X 9 ,X 10 ,X 11 ，X 12 ，X 13 ，X 14 ，X 13 ，X 16 and X 17 Each independently is an amino acid residue, wherein:
X 2 comprising a side chain containing an acidic or polar group;
X 5 comprising a side chain containing an acidic or polar group;
X 13 comprising side chains containing optionally substituted aromatic groups, and
X 1 、X 3 、X 4 、X 7 、X 10 、X 11 and X 14 Each independently is an amino acid residue suitable for stapling, or each independently is stapling.
315. The agent of any one of the preceding embodiments, comprising three or more staple-like structures within 10 to 20 amino acid residues.
316. The agent of any one of the preceding embodiments, comprising three or more staple-like structures within 10 to 15 amino acid residues.
317. The agent of any one of the preceding embodiments, comprising three or more staple-like structures within 15 amino acid residues.
318. The agent of any one of the preceding embodiments, comprising three or more staple-like structures within 14 amino acid residues.
319. The agent of any one of the preceding embodiments, comprising three or more staple-like structures within 11 amino acid residues.
320. The agent of any one of the preceding embodiments, wherein there are three staple-like structures in the peptide.
321. The agent of any one of embodiments 1 to 319, wherein there are four staple-like structures in the peptide.
322. The agent of any one of the preceding embodiments, wherein X 0 、X 1 、X 3 、X 4 、X 7 、X 10 、X 11 And X 14 Each independently an amino acid residue suitable for stapling, or each independently stapling.
323. The agent of any one of the preceding embodiments, wherein X 0 、X 1 、X 3 、X 4 、X 7 、X 10 、X 11 And X 14 Each independently is an amino acid residue suitable for stapling, or each independently is stapling.
324. The agent of any one of the preceding embodiments, wherein X 0 、X 1 、X 3 、X 4 、X 7 、X 10 、X 11 And X 14 Each independently is an amino acid residue suitable for stapling, or each independently is stapling.
325. The agent of any one of the preceding embodiments, wherein X 1 、X 3 、X 4 、X 7 、X 10 、X 11 And X 14 Each independently an amino acid residue suitable for stapling, or each independently stapling.
326. The agent of any one of the preceding embodiments, wherein X 1 、X 3 、X 4 、X 7 、X 10 、X 11 And X 14 Each independently is an amino acid residue suitable for stapling, or each independently is stapling.
327. The agent of any one of the preceding embodiments, wherein X 1 、X 3 、X 4 、X 7 、X 10 、X 11 And X 14 Each independently is an amino acid residue suitable for stapling, or each independently is stapling.
328. The agent of any one of the preceding embodiments, wherein X 0 And X 4 Each independently is an amino acid residue suitable for stapling.
329. The agent of any one of embodiments 1 to 327, wherein X 0 And X 4 Connected by a staple-like structure.
330. The agent of any one of the preceding embodiments, wherein X 1 And X 4 Each independently is an amino acid residue suitable for stapling.
331. The agent of any one of embodiments 1 to 327, wherein X 1 And X 4 Connected by a staple-like structure.
332. Any one of embodiments 1 to 327The medicament, wherein X 1 And X 3 Each independently is an amino acid residue suitable for stapling.
333. The agent of any one of embodiments 1 to 327, wherein X 1 And X 3 Connected by a staple-like structure.
334. The agent of any one of the preceding embodiments, wherein X 4 And X 11 Each independently is an amino acid residue suitable for stapling.
335. The agent of any one of embodiments 1 to 333, wherein X 4 And X 11 Connected by a staple-like structure.
336. The medicament of embodiment 309, wherein X 1 、X 4 And X 11 Each independently is an amino acid residue suitable for stapling.
337. The medicament of embodiment 309, wherein X 1 And X 4 Connected by staple-like structure, and X 4 And X 11 Connected by a staple-like structure.
338. The agent of any one of embodiments 1 to 308, wherein the agent is the agent of any one of embodiments 309 to 337.
339. The agent of any one of the preceding embodiments, wherein X 10 And X 14 Each independently is an amino acid residue suitable for stapling.
340. The agent of any one of embodiments 1 to 337, wherein X 10 And X 14 Connected by a staple-like structure.
341. The agent of any one of the preceding embodiments, wherein X 7 And X 10 Each independently is an amino acid residue suitable for stapling.
342. The agent of any one of embodiments 1 to 340, wherein X 7 And X 10 Connected by a staple-like structure.
343. The agent of any one of the preceding embodiments, wherein X 7 And X 14 Each independently is an amino acid residue suitable for stapling.
344. The agent of any one of embodiments 1 to 342, wherein X 7 And X 14 Connected by a staple-like structure.
345. The agent of any one of embodiments 1 to 331 and 334 to 340, wherein X 3 And X 7 Each independently is an amino acid residue suitable for stapling.
346. The agent of any one of embodiments 1 to 331 and 334 to 340, wherein X 3 And X 7 Connected by a staple-like structure.
347. The agent of any one of the preceding embodiments, wherein the agent comprises an N-terminal group.
348. The agent of any one of the preceding embodiments, wherein the N-terminal group is an acyl group.
349. The medicament of embodiment 347, wherein the N-terminal group comprises a moiety for stapling.
350. The medicament of embodiment 347, wherein the N-terminal group comprises a terminal alkene.
351. The agent of any one of the preceding embodiments, wherein the agent comprises an N-terminal group that is
Ac, NPyroR3,5 hexenyl, 4 pentenyl, bua, C3a, cpc, cbc, cypCO, bnc, CF3CO,2PyCypCO,4THPCO, isobutyryl, ts,15pyraPy,2PyBu,4PymCO,4PyPrpc,3IAPAc,4MePipzPrpC, mePipAc, meImid4SO2, bzAm2OAllyl, hex,2PyzCO,3Phc3, meOPr, lithocholic acid, 2FPhc, phC, meSO2, isovaleryl, etHNCO, tzPyr,8IAP,3PydCO,2PymCO,5PymCO,1Imidac,2F2PyAc,2IAPAc,124TriPr,6QuiAc,3PyAc,123TriAc,1 pyrazole Ac,3PyPrpc,5PymAc,1PydoneAc, 124Ac, me2NAc,8QuiSO2, mPEG4, mPEG8, mPEG16, or mPEG24.
352. The agent of embodiment 347, wherein the N-terminal group is Ac.
353. The agent of any one of the preceding embodiments, wherein X 1 Is an amino acid residue having the structure of formula A-I, A-II or A-III, wherein R a1 And R is a3 Together with intervening atoms to formOptionally substituted 3-to 10-membered rings having 0 to 5 heteroatoms in addition to the intervening atoms.
354. The agent of any one of the preceding embodiments, wherein X 1 is-N (R) a1 )-L a1 -C(-L a -R SP1 )(R a3 )-L a2 -C(O)-。
355. The agent of embodiment 354 wherein R a1 is-H.
356. The agent of any one of embodiments 354 to 355, wherein R a3 is-H.
357. The agent of any one of embodiments 354 to 355, wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
358. The agent of any one of the preceding embodiments, wherein X 1 Is N (R) a1 )(-L a -R SP1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)-。
359. The agent of embodiment 358 wherein R a2 is-H.
360. The agent of embodiment 358 wherein R a2 Is optionally substituted C 1-6 Aliphatic series.
361. The agent of embodiment 358 wherein R a2 Is methyl.
362. The agent of embodiments 354 and 358 through 361, wherein R a1 And R is a3 Taken together with intervening atoms form an optionally substituted 3-to 10-membered ring having 0 to 3 heteroatoms in addition to the intervening atoms.
363. The agent of embodiment 362, wherein R a1 And R is a3 Taken together with intervening atoms to form a member other than R a1 A 5 membered saturated ring having no heteroatoms other than the nitrogen to which it is attached.
364. The agent of any one of embodiments 354 to 363, wherein L a1 Is a covalent bond.
365. The agent of any one of embodiments 354 to 364, wherein L a Is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, itOptionally and independently represented by-O-, -S-, -Cy-, -and-N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
366. The agent of any one of embodiments 354 to 364, wherein L a Is an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
367. The agent of any one of embodiments 354 to 364, wherein L a Is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
368. The agent of any one of embodiments 354 to 367, wherein L a Is optionally substituted- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
369. The agent of any one of embodiments 354 to 368, wherein L a Is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
370. The agent of any one of embodiments 354 to 369, wherein L a2 Is a covalent bond.
371. The agent of any one of embodiments 354 to 370, wherein R SP1 Is optionally substituted-ch=ch 2 。
372. The agent of any one of embodiments 354 to 370, wherein R SP1 is-CH=CH 2 。
373. The agent of any one of embodiments 354 to 370, wherein R SP1 is-COOH.
374. The agent of any one of embodiments 354 to 370, wherein R SP1 Is or comprises an amino group.
375. The agent of any one of embodiments 354 to 370, wherein R SP1 is-NHR wherein R is hydrogen or optionally substituted C 1-6 Aliphatic series.
376. The agent of any one of embodiments 354 to 370, whereinR SP1 is-NHR, wherein R is C 1-6 An alkyl group.
377. The agent of any one of embodiments 354 to 370, wherein R SP1 is-NH 2 Wherein R is C 1-6 An alkyl group.
378. The agent of any one of embodiments 354 to 370, wherein R SP1 is-N 3 。
379. The agent of any one of embodiments 354 to 370, wherein R SP1 Is a terminal or activated alkyne.
380. The agent of any one of embodiments 354 to 370, wherein R SP1 is-C.ident.CH.
381. The agent of any one of embodiments 354 to 370, wherein R SP1 is-SH.
382. The agent of any one of embodiments 1 to 352 wherein X 1 Is PL3, S5, mePro, asp, S6, pro, ala, ser, thioPro, gly, NMebAla, tfeGA, or Asn.
383. The agent of any one of embodiments 1 to 352 wherein X 1 Is that
Ac-PL3, ac-S5, NPyroR3-Asp, ac-MePro,5 hexenyl-MePro, AC-S6,4 pentenyl-MePro, ac-Pro, ac-Ala, bua-PL3, C3a-PL3, cpc-PL3, cbc-PL3, cypCO-PL3,4THPCO-PL3, isobutyryl-PL 3, ac-Asp, ac-Ser, ts-PL3, 15pyraPy-PL3,2PyBu-PL3,4PymCO-PL3,4 pentenyl-ThioPro, 4PyPrpc-PL3,3IAPAc-PL3,4 MePizPrpC-PL 3, mePipAc-PL3, meid 4SO2-PL3, bzam2OAllyl-MePro, ac-Sar, mebAc-Ala, hex-3, NMzCO-3, phc-3, meOPr-PL3, lithocholic acid-PL 3,2FPhc-PL3, phC-PL3, meSO2-PL3, isovaleryl-PL 3, etONCO-PL 3, tzpyr-PL3,8IAP-PL3,3PydCO-PL3,2PymCO-PL3,5PymCO-PL3,1Imidac-PL3,2F2PyAc-PL3,2IAPAc-PL3, 124TriPr-PL3,6QuiAc-PL3,3PyAc-PL3, 123TriAc-PL3,1 pyrazole Ac-PL3,3PyPrpc-PL3,5PymAc-PL3,1 pyrone Ac-PL3, 124TriAc-PL3, me2NAc-PL3,8QuiSO2-PL3, mPEG4-PL3, mPEG8-PL3, mPEG16-PL3, mPEG24-PL3, asro-R3 or NPr-NPyR 3.
384. Any one of embodiments 1 to 352Wherein X is 1 Is PL3, [ 4-pentenyl ] ]MePro, [5 hexenyl ]]MEPro, or [ BzAm2OAllyl ]]MePro。
385. The agent of any one of embodiments 1 to 352 wherein X 1 Is PL3.
386. The agent of any one of embodiments 1 to 352 wherein X 1 Is [ 4-pentenyl ]]MePro, or [5 hexenyl ]]MePro。
387. The agent of any one of the preceding embodiments, wherein X 1 Interact with Va1349 of β -catenin or with its corresponding amino acid residue.
388. The agent of any one of the preceding embodiments, wherein X 3 Is an amino acid residue comprising a carboxyl group.
389. The agent of any one of the preceding embodiments, wherein X 3 Is an amino acid residue having the structure of formula A-I, A-II or A-III, wherein R a2 Or R is a3 Comprising a carboxyl group.
390. The agent of any one of the preceding embodiments, wherein R a2 Or R is a3 is-L a -CO 2 R。
391. The agent of any one of the preceding embodiments, wherein X 3 Is GlnR.
392. The agent of any one of embodiments 1 to 387, wherein X 3 Is the residue of an amino acid comprising an alkene.
393. The agent of any one of embodiments 1 to 387 and 392, wherein X 3 Is comprised of-CH=CH 2 Residues of amino acids of (a).
394. The agent of any one of embodiments 1 through 387 and 392 through 393, wherein X 3 Is comprised of-CH=CH 2 And forms a staple-like structure with another amino acid residue by olefin metathesis.
395. The agent of any one of embodiments 1 to 387, wherein X 3 Is a residue of an amino acid having the structure of formula A-I, A-II or A-III, wherein R a2 Or R is a3 Comprising olefins.
396. Any one of embodiments 1 through 387 and 395The medicament, wherein X 3 Is an amino acid residue having the structure of formula A-I, A-II or A-III, wherein R a2 Or R is a3 is-L a CH＝CH 2 。
397. The agent of any one of the preceding embodiments, wherein X 3 is-N (R) a1 )-L a1 -C(-L a -R SP1 )(R a3 )-L a2 -C(O)-。
398. The agent of embodiment 397, wherein R a1 is-H.
399. The agent of any one of embodiments 397 to 398, wherein R a3 is-H.
400. The agent of any one of embodiments 397 to 398, wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
401. The agent of any one of embodiments 397 to 400, wherein L a1 Is a covalent bond.
402. The agent of any one of embodiments 397 to 401, wherein L a Is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
403. The agent of any one of embodiments 397 to 401, wherein L a Is an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
404. The agent of any one of embodiments 397 to 401, wherein L a Is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
405. The agent of any one of embodiments 397 to 402, wherein L a Is optionally substituted- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
406. Embodiment 397The agent of any one of claims 402, wherein L a Is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
407. The agent of any one of embodiments 397 to 406, wherein L a2 Is a covalent bond.
408. The agent of any one of embodiments 397 to 407, wherein R SP1 Is optionally substituted-ch=ch 2 。
409. The agent of any one of embodiments 397 to 407, wherein R SP1 is-CH=CH 2 。
410. The agent of any one of embodiments 397 to 407, wherein R SP1 is-COOH.
411. The agent of any one of embodiments 397 to 407, wherein R SP1 Is or comprises an amino group.
412. The agent of any one of embodiments 397 to 407, wherein R SP1 is-NHR wherein R is hydrogen or optionally substituted C 1-6 Aliphatic series.
413. The agent of any one of embodiments 397 to 407, wherein R SP1 is-NHR, wherein R is C 1-6 An alkyl group.
414. The agent of any one of embodiments 397 to 407, wherein R SP1 is-NH 2 。
415. The agent of any one of embodiments 397 to 407, wherein R SP1 is-N 3 。
416. The agent of any one of embodiments 397 to 407, wherein R SP1 Is a terminal or activated alkyne.
417. The agent of any one of embodiments 397 to 407, wherein R SP1 is-C.ident.CH.
418. The agent of any one of embodiments 397 to 407, wherein R SP1 is-SH.
419. The agent of any one of embodiments 1 to 387 and 392 to 396, wherein X 3 Is AllilGly, [ Bn ]][ allyl group]Dap、[Phc][ allyl group]Dap、[Piv][ allyl group]Dap or [ CyCO ]][ allyl group]Dap。
420. The agent of any one of the preceding embodiments, wherein X 4 Is an amino acid residue comprising an alkene.
421. The agent of any one of the preceding embodiments, wherein X 4 Is comprised of-CH=CH 2 Amino acid residues of (a) are present.
422. The agent of any one of the preceding embodiments, wherein X 4 Is comprised of-CH=CH 2 And forms a staple-like structure with another amino acid residue by olefin metathesis.
423. The agent of any one of the preceding embodiments, wherein X 4 Is an amino acid residue having the structure of formula A-I, A-II or A-III, wherein R a2 Or R is a3 Comprising olefins.
424. The agent of any one of the preceding embodiments, wherein X 4 Is an amino acid residue having the structure of formula A-I, A-II or A-III, wherein R a2 Or R is a3 is-L a -CH＝CH 2 。
425. The agent of any one of the preceding embodiments, wherein X 4 Is R5, R4 or R6.
426. The agent of any one of embodiments 1 to 419, wherein X 4 Is the residue of an amino acid comprising two olefins.
427. The agent of any one of embodiments 1 to 419 and 426 wherein X 4 Is comprised of two-ch=ch 2 Amino acid residues of (a) are present.
428. The agent of any one of embodiments 1 to 419 and 426 to 427 wherein X 4 Is comprised of two-ch=ch 2 And each forms a staple-like structure with another amino acid residue by olefin metathesis.
429. The agent of any one of embodiments 1 to 419, wherein X 4 Is an amino acid residue having the structure of formula A-I, A-II or A-III, wherein R a2 And R is a3 Each independently comprising an olefin.
430. The agent of any one of embodiments 1 to 419 and 429 wherein X 4 Is of the formula A-I, A-II or A-IIIResidues of amino acids, wherein R a2 And R is a3 Each independently is-L a -CH＝CH 2 。
431. The agent of any one of the preceding embodiments, wherein X 4 is-N (R) a1 )-L a1 -C(-L a -R SP1 )(R a3 )-L a2 -C(O)-。
432. The agent of embodiment 431, wherein R a1 is-H.
433. The agent of any one of embodiments 431 to 432, wherein R a3 is-H.
434. The agent of any one of embodiments 431 to 432, wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
435. The agent of any one of the preceding embodiments, wherein X 4 is-N (R) a1 )-L a1 -C(-L a -R SP1 )(-L a -R SP2 )-L a2 -C(O)-。
436. The agent of any one of embodiments 431 to 435, wherein L a1 Is a covalent bond.
437. The agent of any one of embodiments 431 to 436, wherein L a Is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
438. The agent of any one of embodiments 431 to 436, wherein L a Is an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
439. The agent of any one of embodiments 431 to 436, wherein L a Is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
440. In embodiments 431 to 437The agent of any one of claims, wherein R is attached to SP1 Bonded L a Is optionally substituted- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
441. The agent of any one of embodiments 431 to 437, wherein with R SP1 Bonded L a Is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
442. The agent of any one of embodiments 431 to 441, wherein L a2 Is a covalent bond.
443. The agent of any one of embodiments 431 to 442, wherein R SP1 Is optionally substituted-ch=ch 2 。
444. The agent of any one of embodiments 431 to 442, wherein R SP1 is-CH=CH 2 。
445. The agent of any one of embodiments 431 to 442, wherein R SP1 is-COOH.
446. The agent of any one of embodiments 431 to 442, wherein R SP1 Is or comprises an amino group.
447. The agent of any one of embodiments 431 to 442, wherein R SP1 is-NHR wherein R is hydrogen or optionally substituted C 1-6 Aliphatic series.
448. The agent of any one of embodiments 431 to 442, wherein R SP1 is-NHR, wherein R is C 1-6 An alkyl group.
449. The agent of any one of embodiments 431 to 442, wherein R SP1 is-NH 2 。
450. The agent of any one of embodiments 431 to 442, wherein R SP1 is-N 3 。
451. The agent of any one of embodiments 431 to 442, wherein R SP1 Is a terminal or activated alkyne.
452. The agent of any one of embodiments 431 to 442, wherein R SP1 is-C.ident.CH.
453. The agent of any one of embodiments 431 to 442, wherein R SP1 is-SH.
454. The agent of any one of embodiments 431 to 453, wherein R SP2 Is optionally substituted-ch=ch 2 。
455. The agent of any one of embodiments 431 to 453, wherein R SP2 is-CH=CH 2 。
456. The agent of any one of embodiments 431 to 453, wherein R SP2 is-COOH.
457. The agent of any one of embodiments 431 to 453, wherein R SP2 Is or comprises an amino group.
458. The agent of any one of embodiments 431 to 453, wherein R SP2 is-NHR wherein R is hydrogen or optionally substituted C 1-6 Aliphatic series.
459. The agent of any one of embodiments 431 to 453, wherein R SP2 is-NHR, wherein R is C 1-6 An alkyl group.
460. The agent of any one of embodiments 431 to 453, wherein R SP2 is-NH 2 。
461. The agent of any one of embodiments 431 to 453, wherein R SP2 is-N 3 。
462. The agent of any one of embodiments 431 to 453, wherein R SP2 Is a terminal or activated alkyne.
463. The agent of any one of embodiments 431 to 453, wherein R SP2 is-C.ident.CH.
464. The agent of any one of embodiments 431 to 453, wherein R SP2 is-SH.
465. The agent of any one of embodiments 431 to 464 wherein R is attached to SP1 Bonded L a Is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
466. The agent of any one of embodiments 431 to 464 wherein R is attached to SP1 Bonded L a Is optionally substituted diValence C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
467. The agent of any one of embodiments 431 to 464 wherein R is attached to SP1 Bonded L a Is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
468. The agent of any one of embodiments 431 to 464 wherein R is attached to SP1 Bonded L a Is optionally substituted- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
469. The agent of any one of embodiments 431 to 464 wherein R is attached to SP1 Bonded L a Is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
470. The agent of any one of embodiments 435 to 469, wherein R is attached to SP2 Bonded L a Is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
471. The agent of any one of embodiments 435 to 469, wherein R is attached to SP2 Bonded L a Is an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
472. The agent of any one of embodiments 435 to 469, wherein R is attached to SP2 Bonded L a Is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R '), -C (O) -, -C (O) N (R '), -N (R ') C (O) O-substitution.
473. The agent of any one of embodiments 435 to 469, wherein R is attached to SP2 Bonded L a Is optionally substituted- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
474. The agent of any one of embodiments 435 to 469, wherein R is attached to SP2 Bonded L a Is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
475. The agent of any one of embodiments 1 to 419 and 426 to 430 wherein X 4 Is B5.
476. The agent of any one of embodiments 1 to 419, wherein X 4 Is B5, npg, asp, R5, ile, ala, cha, chg, ser, leu, R4, R6, phe, or S5.
477. The agent of any one of the preceding embodiments, wherein X 7 Is an amino acid residue comprising an optionally substituted carboxy group, an optionally substituted amino group or an azido group.
478. The agent of any one of the preceding embodiments, wherein X 7 Is a residue of an amino acid having the structure of formula A-I, A-II or A-III, wherein R a2 Or R is a3 Comprising carboxyl, amino or azido groups.
479. The agent of any one of the preceding embodiments, wherein X 7 Is a residue of an amino acid having the structure of formula A-I, A-II or A-III, wherein R a2 Or R is a3 is-L a -CO 2 R,-L a -N 3 or-L a -L-R。
480. The agent of any one of the preceding embodiments, wherein X 7 is-N (R) a1 )-L a1 -C(-L a -R SP1 )(R a3 )-L a2 -C(O)-。
481. The agent of embodiment 480 wherein R a1 is-H.
482. The agent of any one of embodiments 480 to 481, wherein R a3 is-H.
483. The agent of any one of embodiments 480 to 481, wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
484. The agent of any one of embodiments 480 to 483, wherein L a1 Is a covalent bond.
485. The agent of any one of embodiments 480 to 484, wherein L a Is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
486. The agent of any one of embodiments 480 to 485, wherein L a Is optionally substituted- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
487. The agent of any one of embodiments 480 to 486, wherein L a Is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
488. The agent of any one of embodiments 480 to 487, wherein L a2 Is a covalent bond.
489. The agent of any one of embodiments 480 to 488, wherein R SP1 Is optionally substituted-ch=ch 2 。
490. The agent of any one of embodiments 480 to 489, wherein R SP1 is-CH=CH 2 。
491. The agent of any one of embodiments 480 to 488, wherein R SP1 is-COOH.
492. The agent of any one of embodiments 480 to 488, wherein R SP1 Is or comprises an amino group.
493. The agent of any one of embodiments 480 to 488, wherein R SP1 is-NHR wherein R is hydrogen or optionally substituted C 1-6 Aliphatic series.
494. The agent of any one of embodiments 480 to 488, wherein R SP1 is-NHR, wherein R is C 1-6 An alkyl group.
495. The agent of any one of embodiments 480 to 488, wherein R SP1 is-NH 2 。
496. The agent of any one of embodiments 480 to 488, wherein R SP1 is-N 3 。
497. The agent of any one of embodiments 480 to 488, wherein R SP1 Is a terminal or activated alkyne.
498. The agent of any one of embodiments 480 to 488, wherein R SP1 is-C.ident.CH.
499. The agent of any one of embodiments 480 to 488, wherein R SP1 is-SH.
500. The agent of any one of the preceding embodiments, wherein X 7 Is GlnR, lys, [29N2 spiro undecane ]GlnR, [4 aminopiperidine]GlnR, sAla, triAzLys, [ isophthalate ]]Lys, [ succinic acid ester ]]Lys、[Me2Mal]Lys, [ dibenzoate ]]Lys or [ biphenyl 33COOH]Lys。
501. The agent of any one of the preceding embodiments, wherein X 7 Is GlnR, [29N2 spiroundecane]GlnR or [4 aminopiperidine]GlnR。
502. The agent of any one of embodiments 1 to 500, wherein X 7 Is Lys.
503. The agent of any one of embodiments 1 to 500, wherein X 7 Is TriAzLys.
504. The agent of any one of the preceding embodiments, wherein X 10 Is a residue comprising the amino acid: optionally substituted carboxy, optionally substituted amino, azido, optionally substituted alkynyl or optionally substituted thiol.
505. The agent of any one of the preceding embodiments, wherein X 10 Is a residue of an amino acid having the structure of formula A-I, A-II or A-III, wherein R a2 Or R is a3 Comprising carboxyl, amino, azido, alkynyl or thiol groups.
506. The agent of any one of the preceding embodiments, wherein X 10 Is a residue of an amino acid having the structure of formula A-I, A-II or A-III, wherein R a2 Or R is a3 is-L a -CO 2 R,-L a -N 3 or-L a -L-R。
507. The agent of any one of the preceding embodiments, wherein X 10 is-N (R) a1 )-L a1 -C(-L a -R SP1 )(R a3 )-L a2 -C(O)-。
508. The drug of embodiment 507An agent, wherein R is a1 is-H.
509. The agent of any one of embodiments 507-508 wherein R a3 is-H.
510. The agent of any one of embodiments 507-508 wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
511. The agent of any one of embodiments 507-510 wherein L a1 Is a covalent bond.
512. The agent of any one of embodiments 507-511 wherein L a Is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
513. The agent of any one of embodiments 507-512 wherein L a Is optionally substituted- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
514. The agent of any one of embodiments 507-513 wherein L a Is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
515. The agent of any one of embodiments 507-514 wherein L a2 Is a covalent bond.
516. The agent of any one of embodiments 507-515 wherein R SP1 Is optionally substituted-ch=ch 2 。
517. The agent of any one of embodiments 507-515 wherein R SP1 is-CH=CH 2 。
518. The agent of any one of embodiments 507-515 wherein R SP1 is-COOH.
519. The agent of any one of embodiments 507-515 wherein R SP1 Is or comprises an amino group.
520. The agent of any one of embodiments 507-515 wherein R SP1 is-NHR wherein R is hydrogen or optionally substituted C 1-6 Aliphatic series.
521. The agent of any one of embodiments 507-515 wherein R SP1 is-NHR, wherein R is C 1-6 An alkyl group.
522. The agent of any one of embodiments 507-515 wherein R SP1 is-NH 2 。
523. The agent of any one of embodiments 507-515 wherein R SP1 is-N 3 。
524. The agent of any one of embodiments 507-515 wherein R SP1 Is a terminal or activated alkyne.
525. The agent of any one of embodiments 507-515 wherein R SP1 is-C.ident.CH.
526. The agent of any one of embodiments 507-515 wherein R SP1 is-SH.
527. The agent of any one of the preceding embodiments, wherein X 10 Is that
Lys, glnR, triAzLys, sla, dLys, asnR, hGlnR, iPrLys, triAzOrn, DGlnR, orn,4pipa, sch2s, [8fbb ] Cys, [4fb ] Cys, [ mXyl ] Cys, [ ohyl ] Cys, [ pXyl ] Cys, dlan, dDab, NMeOrn, [2_6-naph ] Cys, or [3_3-biph ] Cys.
528. The agent of any one of the preceding embodiments, wherein X 10 Is Lys, glnR or TriAzLys.
529. The agent of any one of embodiments 1 to 528, wherein X 10 Is Lys.
530. The agent of any one of embodiments 1 to 528, wherein X 10 Is GlnR.
531. The agent of any one of embodiments 1 to 528, wherein X 10 Is TriAzLys.
532. The agent of any one of the preceding embodiments, wherein X 11 Is the residue of an amino acid comprising an alkene.
533. The agent of any one of the preceding embodiments, wherein X 11 Is comprised of-CH=CH 2 Residues of amino acids of (a).
534. The agent of any one of the preceding embodiments, wherein X 11 Is comprised of-CH=CH 2 Residues of amino acids of (2) and by olefinsMetathesis forms a staple-like structure with another amino acid residue.
535. The agent of any one of the preceding embodiments, wherein X 11 Is a residue of an amino acid having the structure of formula A-I, A-II or A-III, wherein R a2 Or R is a3 Comprising olefins.
536. The agent of any one of the preceding embodiments, wherein X 11 Is a residue of an amino acid having the structure of formula A-I, A-II or A-III, wherein R a2 Or R is a3 is-L a -CH＝CH 2 。
537. The agent of any one of the preceding embodiments, wherein X 11 is-N (R) a1 )-L a1 -C(-L a -R SP1 )(R a3 )-L a2 -C(O)-。
538. The agent of embodiment 537 wherein R a1 is-H.
539. The agent of any one of embodiments 537-538, wherein L a1 Is a covalent bond.
540. The agent of any one of embodiments 537-539 wherein L a Is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
541. The agent of any one of embodiments 537-539 wherein L a Is an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
542. The agent of any one of embodiments 537-539 wherein L a Is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
543. The agent of any one of embodiments 537-540, wherein L a Is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
544. The agent of any one of embodiments 537-540, wherein L a2 Is a covalent bond.
545. The agent of any one of embodiments 537-544, wherein R SP1 Is optionally substituted-ch=ch 2 。
546. The agent of any one of embodiments 537-544, wherein R SP1 is-CH=CH 2 。
547. The agent of any one of embodiments 537-544, wherein R SP1 is-COOH.
548. The agent of any one of embodiments 537-544, wherein R SP1 Is or comprises an amino group.
549. The agent of any one of embodiments 537-544, wherein R SP1 is-NHR wherein R is hydrogen or optionally substituted C 1-6 Aliphatic series.
550. The agent of any one of embodiments 537-544, wherein R SP1 is-NHR, wherein R is C 1-6 An alkyl group.
551. The agent of any one of embodiments 537-544, wherein R SP1 is-NH 2 。
552. The agent of any one of embodiments 537-544, wherein R SP1 is-N 3 。
553. The agent of any one of embodiments 537-544, wherein R SP1 Is a terminal or activated alkyne.
554. The agent of any one of embodiments 537-544, wherein R SP1 is-C.ident.CH.
555. The agent of any one of embodiments 537-544, wherein R SP1 is-SH.
556. The agent of any one of embodiments 537-555 wherein one methylene unit of L is replaced by-N (R') -.
557. The agent of any one of embodiments 537-555 wherein one methylene unit of L is replaced by-N (R') C (O) O-.
558. The agent of any one of embodiments 556 to 557 wherein R' is-H.
559. The agent of any of embodiments 556-557 wherein R' is C 1-6 Aliphatic series.
560. The agent of any one of embodiments 556 to 557 wherein R' and R a3 Taken together with intervening atoms, form an optionally substituted 3-to 14-membered ring having 0 to 5 heteroatoms in addition to the nitrogen atom to which R' is attached.
561. The agent of any one of embodiments 556 to 557 wherein R' and R a3 Taken together with intervening atoms, form an optionally substituted 3-to 8-membered ring having 0 to 5 heteroatoms in addition to the nitrogen atom to which R' is attached.
562. The agent of any one of embodiments 556 to 557 wherein R' and R a3 Taken together with intervening atoms, form an optionally substituted 3-to 7-membered ring having no heteroatoms other than the nitrogen atom to which R' is attached.
563. The agent of any one of embodiments 560 to 562, wherein the ring is monocyclic.
564. The agent of any one of embodiments 560 to 563 wherein the ring is saturated.
565. The agent of any one of embodiments 560 to 564, wherein the ring is 5-membered.
566. The agent of any one of embodiments 537-559, wherein R a3 is-H.
567. The agent of any one of embodiments 537-559, wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
568. The agent of any one of embodiments 537-555 and 566-567, wherein L a Is optionally substituted- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
569. The agent of embodiment 568 wherein L a Is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
570. The agent of any one of the preceding embodiments, wherein X 11 Is that
PyrS2, lys,3thi, ala, phe, SPIP3, pyrSadNap 3 butene, SPIP2, az3, dapAc7EDA, leu,3 allyloxy PyrSa, pyrSaV3 butene, az2, pyrS1, pyrSc72SMe3ROMe, pyrSc72RMe3SOMe, pyrSc7O45RMe, pyrSc7O45SMe, pyrSc73Me2, pyrSc7, pyrSaA3 butene, pyrSadA3 butene, dap7Gly, dap7Pent, dapAc7PDA, dap7Abu,4 VinyPyrSasa, pyrSadV3 butene, pyrSar 3 butene, pyrSap 3 butene, pyrPro 3 butene, pyrSap 2 Phase3, or PySala 3 Ac.
571. The agent of any one of the preceding embodiments, wherein X 11 Is pyrS2.
572. The agent of any one of the preceding embodiments, wherein X 14 Is the residue of an amino acid comprising a carboxyl, amino, azido, alkynyl or thiol group.
573. The agent of any one of the preceding embodiments, wherein X 14 Is a residue of an amino acid having the structure of formula A-I, A-II or A-III, wherein R a2 Or R is a3 Comprising carboxyl, amino, azido, alkynyl or thiol groups.
574. The agent of any one of the preceding embodiments, wherein X 14 is-N (R) a1 )-L a1 -C(-L a -R SP1 )(R a3 )-L a2 -C(O)-。
575. The agent of embodiment 574, wherein R a1 is-H.
576. The agent of embodiments 574 to 575, wherein R a3 is-H.
577. The agent of embodiments 574 to 575, wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
578. The agent of embodiments 574 to 577, wherein L a1 Is a covalent bond.
579. The agent of embodiments 574 to 578, wherein L a Is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
580. Implementation prescriptionThe pharmaceutical preparation of cases 574 to 578, wherein L a Is an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
581. The agent of embodiments 574 to 578, wherein L a Is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
582. The agent of embodiments 574 to 579, wherein L a Is optionally substituted- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
583. The agent of embodiments 574 to 579, wherein L a Is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
584. The agent of embodiments 574 to 583, wherein L a2 Is a covalent bond.
585. The agent of embodiments 574 to 584, wherein R SP1 Is optionally substituted-ch=ch 2 。
586. The agent of embodiments 574 to 584, wherein R SP1 is-CH=CH 2 。
587. The agent of embodiments 574 to 584, wherein R SP1 is-COOH.
588. The agent of embodiments 574 to 584, wherein R SP1 Is or comprises an amino group.
589. The agent of embodiments 574 to 584, wherein R SP1 is-NHR wherein R is hydrogen or optionally substituted C 1-6 Aliphatic series.
590. The agent of embodiments 574 to 584, wherein R SP1 is-NHR, wherein R is C 1-6 An alkyl group.
591. The agent of embodiments 574 to 584, wherein R SP1 is-NH 2 。
592. The agent of embodiments 574 to 584, wherein R SP1 is-N 3 。
593. The agent of embodiments 574 to 584, wherein R SP1 Is a terminal or activated alkyne.
594. The agent of embodiments 574 to 584, wherein R SP1 is-C.ident.CH.
595. The agent of embodiments 574 to 584, wherein R SP1 is-SH.
596. The agent of any one of the preceding embodiments, wherein X 14 Is a residue of an amino acid having the structure of formula A-I, A-II or A-III, wherein R a2 Or R is a3 is-L a -CO 2 R，-L a -N 3 or-L a -L-R。
597. The agent of any one of the preceding embodiments, wherein X 14 Is GlnR, lys, sA, gln, cys, triAzLys, asnR, hGlnR,4PipA,sAbu,Orn,GlnR, [4mam piperidine]GlnR, [39N2 spiroundecane]GlnR, [29N2 spiroundecane]GlnR, iPrLys, sCH2S, [ diaminobutane ]]GlnR, [4 aminopiperidine]GlnR、dGlnR。
598. The agent of any one of embodiments 1 to 597, wherein X 14 Is GlnR, lys or sA.
599. The agent of any one of embodiments 1 to 598, wherein X 14 Is GlnR.
600. The agent of any one of embodiments 1 to 598, wherein X 14 Is Lys.
601. The agent of any one of embodiments 1 to 598, wherein X 14 Is sA.
602. The agent of any one of the preceding embodiments, wherein the pairs of amino acid residues suitable for stapling each independently comprise an acidic group.
603. The agent of any one of the preceding embodiments, wherein the pairs of amino acid residues suitable for stapling each independently comprise-COOH or an activated form thereof.
604. The agent of any one of embodiments 602-603, wherein the pair is stapled by reaction with a linking agent that is a diamine or salt thereof.
605. The agent of any one of the preceding embodiments, wherein pairs of amino acid residues suitable for stapling each independently comprise an amino group.
606. The agent of embodiment 605, wherein the pair is stapled by reaction with a linking agent that is a diacid or salt thereof.
607. The medicament of embodiment 605, wherein the pair is stapled by reaction with a linking reagent comprising two-COOHs or salts thereof.
608. The agent of any one of the preceding embodiments, wherein the pairs of amino acid residues suitable for stapling each independently comprise a reactive group, and the reactive group of one is reactive with the other by a cycloaddition reaction.
609. The agent of any one of the preceding embodiments, wherein one of the pair of amino acid residues suitable for stapling comprises-N 3 The other comprises alkynes.
610. The medicament of embodiment 609, wherein the pair is stapled by a click reaction.
611. The agent of any one of the preceding embodiments, wherein the pairs of amino acid residues suitable for stapling each independently comprise a nucleophilic group.
612. The agent of any one of the preceding embodiments, wherein the pairs of amino acid residues suitable for stapling each independently comprise-SH.
613. The agent of any one of embodiments 611 to 612, wherein the pair is stapled by reaction with a linking reagent comprising two leaving groups.
614. The agent of any one of embodiments 611 to 613, wherein the pair of passes has R x -L”-R x The structured linking reagent reacts to become stapled, wherein each R x Independently a leaving group.
615. The agent of any one of embodiments 613 to 614, wherein each leaving group is-Br.
616. The agent of any one of the preceding embodiments, wherein X 10 And X 14 One of which is a residue of an amino acid comprising a carboxyl group and the other is a residue of an amino acid comprising an amino group.
617. The agent of any one of the preceding embodiments, wherein X 10 And X 14 Connected by a staple-like structure, wherein the staple-like structure comprises-C (O) N (R') -.
618. The agent of any one of the preceding embodiments, wherein X 7 And X 10 One of which is a residue of an amino acid comprising a carboxyl group and the other is a residue of an amino acid comprising an amino group.
619. The agent of any one of the preceding embodiments, wherein X 7 And X 10 Connected by a staple-like structure, wherein the staple-like structure comprises-C (O) N (R') -.
620. The agent of any one of the preceding embodiments, wherein X 7 And X 14 One of which is a residue of an amino acid comprising a carboxyl group and the other is a residue of an amino acid comprising an amino group.
621. The agent of any one of the preceding embodiments, wherein X 7 And X 14 Connected by a staple-like structure, wherein the staple-like structure comprises-C (O) N (R') -.
622. The agent of any one of the preceding embodiments, wherein X 3 And X 7 One of which is a residue of an amino acid comprising a carboxyl group and the other is a residue of an amino acid comprising an amino group.
623. The agent of any one of the preceding embodiments, wherein X 3 And X 7 Connected by a staple-like structure, wherein the staple-like structure comprises-C (O) N (R') -.
624. The agent of any one of the preceding embodiments, wherein X 10 And X 14 One of which is a residue of an amino acid comprising an azido group and the other of which is a residue of an amino acid comprising an alkynyl group.
625. The agent of any one of the preceding embodiments, wherein X 10 And X 14 Connected by a staple-like structure, wherein the staple-like structure comprises optionallySubstituted triazolene rings.
626. The agent of any one of the preceding embodiments, wherein X 7 And X 10 One of which is a residue of an amino acid comprising an azido group and the other of which is a residue of an amino acid comprising an alkynyl group.
627. The agent of any one of the preceding embodiments, wherein X 7 And X 10 Is connected by a staple-like structure, wherein the staple-like structure comprises an optionally substituted triazolene ring.
628. The agent of any one of the preceding embodiments, wherein X 10 And X 14 Are residues of amino acids each independently comprising a thiol group.
629. The agent of any one of the preceding embodiments, wherein X 10 And X 14 Connected by a staple-like structure, wherein the staple-like structure comprises-S-Cy-S-.
630. A medicament which is a stapled peptide comprising three staple-like structures, wherein a first staple-like structure and a second staple-like structure are bonded to the same amino acid residue and a third staple-like structure is bonded to two amino acid residues, none of which are bonded to the first staple-like structure or the second staple-like structure.
631. A medicament which is a stapled peptide comprising three staple-like structures, wherein a first staple-like structure and a second staple-like structure are bonded to the same amino acid residue and a third staple-like structure is bonded to two amino acid residues, none of which are bonded to the first staple-like structure or the second staple-like structure.
632. The agent of any one of the preceding embodiments, comprising a polypeptide having L s Staple-like structure of the structure, L s The structure is-L s1 -L s2 -L s3 -。
633. The medicament of any one of the preceding embodiments, comprising three staple-like structures, each independently having L s Structure of the L s The structure is-L s1 -L s2 -L s3 -。
634. The medicament of any of the preceding embodiments, wherein there are three staple-like structures in the medicament, each independently having L s Structure of the L s The structure is-L s1 -L s2 -L s3 -。
635. The medicament of any one of embodiments 1 to 632, comprising four staple-like structures, each independently having L s Structure of the L s The structure is-L s1 -L s2 -L s3 -。
636. The agent of any one of embodiments 1 to 632, wherein there are four staple-like structures in the agent, each independently having L s Structure of the L s The structure is-L s1 -L s2 -L s3 -。
637. The agent of any one of the preceding embodiments, comprising a polypeptide having L s Staple-like structure of the structure, L s The structure is-L s1 -L s2 -L s3 -, wherein the staple-like structure is aligned with X 1 And X 3 And (5) bonding.
638. The agent of any one of the preceding embodiments, comprising a polypeptide having L s Staple-like structure of the structure, L s The structure is-L s1 -L s2 -L s3 -, wherein the staple-like structure is aligned with X 1 And X 4 And (5) bonding.
639. The agent of any one of the preceding embodiments, comprising a polypeptide having L s Staple-like structure of the structure, L s The structure is-L s1 -L s2 -L s3 -, wherein the staple-like structure is aligned with X 4 And X 11 And (5) bonding.
640. The agent of any one of the preceding embodiments, comprising a polypeptide having L s Staple-like structure of the structure, L s The structure is-L s1 -L s2 -L s3 -, wherein the staple-like structure is aligned with X 3 And X 7 And (5) bonding.
641. Any of the foregoing embodimentsThe medicament comprises a compound having L s Staple-like structure of the structure, L s The structure is-L s1 -L s2 -L s3 -, wherein the staple-like structure is aligned with X 7 And X 10 And (5) bonding.
642. The agent of any one of the preceding embodiments, comprising a polypeptide having L s Staple-like structure of the structure, L s The structure is-L s1 -L s2 -L s3 -, wherein the staple-like structure is aligned with X 7 And X 14 And (5) bonding.
643. The agent of any one of the preceding embodiments, comprising a polypeptide having L s Staple-like structure of the structure, L s The structure is-L s1 -L s2 -L s3 -, wherein the staple-like structure is aligned with X 10 And X 14 And (5) bonding.
644. The agent of any one of embodiments 632 to 643, wherein L s1 Is a covalent bond or an optionally substituted divalent linear or branched, saturated or partially unsaturated C 1-10 Hydrocarbon chains in which one or more methylene units are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-，-S-Cy-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
645. The agent of embodiment 644, wherein L s1 Is an optionally substituted divalent linear or branched, saturated or partially unsaturated C 1-10 Hydrocarbon chains in which one or more methylene units are optionally and independently replaced by:
-C(R’) 2 -，-Cy-,-O-，-S，-S-S，-S-Cy-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
646. The agent of embodiment 644, wherein L s1 Is optionally takenSubstituted divalent linear or branched, saturated or partially unsaturated C 1-6 Hydrocarbon chains in which one or more methylene units are optionally and independently replaced by-O-, -Cy-, -S-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-.
647. The agent of embodiment 644, wherein L s1 Is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
648. The agent of any of embodiments 645 through 647, wherein L s1 comprising-N (R') -.
649. The agent of any of embodiments 645 through 647, wherein L s1 comprising-N (R') C (O) O-.
650. The agent of embodiment 649, wherein-N (R') -is closer to L s2 。
651. The agent of embodiment 649, wherein-O-is closer to L s2 。
652. The agent of any of embodiments 645 through 647, wherein L s1 Is- (CH) 2 )m-N(R’)-(CH 2 ) n-, wherein m and n are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.
653. The agent of any of embodiments 645 through 647, wherein L s1 Is- (CH) 2 )m-N(R’)-C(O)-O-(CH 2 ) n-, wherein m and n are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.
654. The agent of any one of embodiments 652 to 653, wherein- (CH) 2 ) m-and L s2 And (5) bonding.
655. The agent of any one of embodiments 652 to 653, wherein- (CH) 2 ) n-and L s2 And (5) bonding.
656. The agent of any one of embodiments 652 to 655, wherein m is 1.
657. The agent of any one of embodiments 652 to 655, wherein m is 2.
658. The agent of any one of embodiments 652 to 657 wherein n is 3.
659. The agent of any one of embodiments 648-658 wherein R' is-H.
660. The agent of any one of embodiments 648-658 wherein R' is optionally substituted C 1-6 Aliphatic series.
661. The agent of any one of embodiments 648-658 wherein R' is methyl.
662. The agent of any one of embodiments 648 to 658 wherein R' and L s1 R of bonded amino acid residue a3 And intervening atoms together form an optionally substituted 3-to 10-membered ring having 0 to 5 heteroatoms in addition to the intervening atoms.
663. The agent of embodiment 662 wherein R' and L s3 R of bonded amino acid residue a3 And intervening atoms together form a 3 to 10 membered monocyclic ring having 0 to 5 heteroatoms in addition to the intervening atoms.
664. The agent of any one of embodiments 662 to 663 wherein the formed ring is saturated.
665. The agent of any one of embodiments 662 to 664 wherein the ring formed is 4-membered.
666. The agent of any one of embodiments 662 to 664 wherein the ring formed is 5-membered.
667. The agent of any one of embodiments 662-666 wherein the formed ring has no heteroatoms other than the intervening atoms.
668. The agent of embodiment 664, wherein L s1 Is optionally substituted- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
669. The agent of embodiment 664, wherein L s1 Is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
670. The agent of embodiment 664, wherein L s1 is-CH 2 -。
671. The agent of embodiment 664, wherein L s1 Is optionally substituted- (CH) 2 ) n-C (O) -, where n is 1, 2, 3, 4,5 or 6.
672. The agent of embodiment 664, wherein L s1 Is- (CH) 2 ) n-C (O) -, wherein n is 1, 2, 3, 4, 5 or 6.
673. The agent of embodiment 664, wherein L s1 Is- (CH) 2 ) n-C (O) -, where n is 2 or 3.
674. The agent of any one of embodiments 632 to 673, wherein L s1 Ratio of AND L s3 -the amino acid residue to which the bond is closer to the N-terminal amino acid residue.
675. The agent of any one of embodiments 632 to 674, wherein L s1 Bonded to carbon atoms of the peptide backbone.
676. The agent of any one of embodiments 632 to 675, wherein L s1 Bonded to the alpha carbon atom of the amino acid residue.
677. The agent of any one of embodiments 632 to 674, wherein L s1 Bonded to the nitrogen atom of the peptide backbone.
678. The agent of any one of embodiments 632 to 674, wherein L s1 Is bonded to a nitrogen atom of the peptide backbone, wherein the nitrogen atom belongs to an amino group bonded to an alpha carbon atom of an amino acid residue.
679. The agent of any of embodiments 677 to 678, wherein said nitrogen atom is attached to L s1 -C (O) -bonding.
680. The agent of any one of embodiments 632 to 679, wherein L s2 Is a covalent bond or an optionally substituted divalent linear or branched, saturated or partially unsaturated C 1-10 Hydrocarbon chains in which one or more methylene units are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-，-S-Cy-S-,-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-,-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
681. The agent of any one of embodiments 632 to 679, wherein L s2 Is an optionally substituted divalent linear or branched,Saturated or partially unsaturated C 1-10 Hydrocarbon chains in which one or more methylene units are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-，-S-Cy-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
682. The agent of embodiment 681, wherein L s2 Is optionally substituted-ch=ch-.
683. The agent of embodiment 681, wherein L s2 Is-ch=ch-.
684. The agent of embodiment 681 wherein the double bond is E.
685. The agent of embodiment 681 wherein the double bond is Z.
686. The agent of embodiment 681, wherein L s2 Is optionally substituted-CH 2 -CH 2 -。
687. The agent of embodiment 681, wherein L s2 is-CH 2 -CH 2 -。
688. The agent of embodiment 681, wherein L s2 is-Cy-.
689. The agent of embodiment 688, wherein-Cy-is an optionally substituted saturated or partially unsaturated 5-to 6-membered ring having 0 to 4 heteroatoms.
690. The agent of embodiment 688 wherein-Cy-is an optionally substituted benzene ring.
691. The agent of embodiment 688, wherein-Cy-is an optionally substituted 5-to 6-membered aromatic ring having 1-4 heteroatoms.
692. The agent of embodiment 688, wherein-Cy-is optionally substituted
693. The agent of embodiment 688, wherein-Cy-is
694. The agent of embodiment 688, wherein-Cy-is optionally substituted
695. The agent of embodiment 688, wherein-Cy-is
696. The agent of any one of embodiments 694-695 wherein carbon atoms are bonded to L s1 And (5) bonding.
697. The agent of any one of embodiments 694-695 wherein carbon atoms are bonded to L s3 And (5) bonding.
698. The agent of embodiment 681, wherein L s2 is-C (O) N (R') -.
699. The agent of embodiment 698, wherein R' is-H.
700. The agent of embodiment 698, wherein R' is optionally substituted C 1-6 Aliphatic series.
701. The agent of any one of embodiments 698 to 700, wherein-N (R') -is identical to L s1 And (5) bonding.
702. The agent of any one of embodiments 698 to 700, wherein-N (R') -is identical to L s3 And (5) bonding.
703. The agent of embodiment 681 wherein one or more methylene units are independently replaced by-C (O) N (R ') -or-N (R ') -and one or more methylene units are independently replaced by-C (R ') 2 -substitution, wherein one or more-C (R') 2 -one or more R ' of (C) N (R ') -or-N (R ') -each independently together with an intervening atom of R ' of-C (O) N (R ') -form an optionally substituted 3-to 10-membered ring having 0 to 5 heteroatoms in addition to the intervening atom.
704. The agent of embodiment 703 wherein the formed ring is saturated.
705. The agent of any one of embodiments 703-704, wherein the formed ring is 4-membered.
706. The agent of any one of embodiments 703-705 wherein the ring formed is 5-membered.
707. The agent of any one of embodiments 703-706, wherein the formed ring has no heteroatoms other than the intervening atoms.
708. The agent of embodiment 681, wherein L s2 is-S-L' -S-.
709. The agent of embodiment 708 wherein L "is a covalent bond or an optionally substituted divalent linear or branched, saturated or partially unsaturated C 1-10 Hydrocarbon chains in which one or more methylene units are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-，-S-Cy-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
710. The agent of embodiment 708 wherein L "is an optionally substituted divalent linear or branched, saturated or partially unsaturated C 1-10 Hydrocarbon chains in which one or more methylene units are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-，-S-Cy-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
711. The agent of embodiment 708 wherein L "is-Cy-or comprises-Cy-.
712. The agent of embodiment 708 wherein L "is- (CH) 2 )m-Cy-(CH 2 ) n-or comprises- (CH) 2 )m-Cy-(CH 2 ) n-, wherein m and n are each optionally substituted 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and each-CH 2 Optionally substituted.
713. The agent of embodiment 712 wherein m and n are each independently 1.
714. The agent of any one of embodiments 711 to 713, wherein is optionally substituted phenyl.
715. The agent of any one of embodiments 711 to 713 wherein is an optionally substituted 5-to 6-membered aromatic ring having 1 to 4 heteroatoms.
716. The agent of any one of embodiments 632 to 715, wherein L s3 Is a covalent bond or an optionally substituted divalent linear or branched, saturated or partially unsaturated C 1-10 Hydrocarbon chains in which one or more methylene units are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-，-S-Cy-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
717. The agent of embodiment 716 wherein L s3 Is an optionally substituted divalent linear or branched, saturated or partially unsaturated C 1-10 Hydrocarbon chains in which one or more methylene units are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-，-S-Cy-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
718. The agent of embodiment 716 wherein L s3 Is an optionally substituted divalent linear or branched, saturated or partially unsaturated C 1-6 Hydrocarbon chains in which one or more methylene units are optionally and independently replaced by-O-, -Cy-, -S-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-.
719. The agent of embodiment 716 wherein L s3 Is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
720. The agent of any one of embodiments 717 to 719, wherein L s3 comprising-N (R') -.
721. The agent of any one of embodiments 717 to 719, wherein L s3 comprising-N (R') C (O) O-.
722. The agent of embodiment 721, wherein-N (R') -is closer to L s2 。
723. The agent of embodiment 721 wherein-O-is closer to L s2 。
724. The agent of any one of embodiments 717 to 719, wherein L s3 Is- (CH) 2 )m-N(R’)-(CH 2 ) n-, wherein m and n are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.
725. The agent of any one of embodiments 717 to 719, wherein L s3 Is- (CH) 2 )m-N(R’)-C(O)-O-(CH 2 ) n-, wherein m and n are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.
726. The agent of any one of embodiments 724 to 725, wherein- (CH) 2 ) n-and L s2 And (5) bonding.
727. The agent of any one of embodiments 724 to 725, wherein- (CH) 2 ) m-and L s2 And (5) bonding.
728. The agent of any one of embodiments 724 to 727, wherein m is 1.
729. The agent of any one of embodiments 724 to 727, wherein m is 2.
730. The agent of any one of embodiments 724 to 729, wherein n is 3.
731. The agent of any one of embodiments 720 to 730, wherein R' is-H.
732. The agent of any one of embodiments 720 through 730, wherein R' is optionally substituted C 1-6 Aliphatic series.
733. The agent of any one of embodiments 720 to 730, wherein R' is methyl.
734. The agent of any one of embodiments 720 through 730, wherein R' and L s3 R of bonded amino acid residue a3 And intervening atoms together form an optionally substituted 3-to 10-membered ring having 0 to 5 heteroatoms in addition to the intervening atoms.
735. The agent of any one of embodiments 720 through 730, wherein R' and L s3 R of bonded amino acid residue a3 And intervening atoms together form a 3 to 10 membered monocyclic ring having 0 to 5 heteroatoms in addition to the intervening atoms.
736. The agent of any one of embodiments 734 to 735, wherein the formed ring is saturated.
737. The agent of any one of embodiments 734 to 736, wherein the ring formed is 4 membered.
738. The agent of any one of embodiments 734 to 737, wherein the ring formed is 5 membered.
739. The agent of any one of embodiments 734 to 738, wherein the formed ring has no heteroatoms other than the intervening atoms.
740. The agent of embodiment 716 wherein L s3 Is optionally substituted- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
741. The agent of embodiment 716 wherein L s3 Is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
742. The agent of embodiment 716 wherein L s3 Is- (CH) 2 ) 3 -。
743. The agent of embodiment 716 wherein L s3 Is- (CH) 2 ) 2 -。
744. The agent of embodiment 716 wherein L s3 is-CH 2 -。
745. The agent of embodiment 716 wherein L s3 Is optionally substituted- (CH) 2 ) n-C (O) -, wherein n is 1, 2, 3, 4, 5 or 6.
746. The agent of embodiment 716 wherein L s3 Is- (CH) 2 ) n-C (O) -, wherein n is 1, 2, 3, 4, 5 or 6.
747. Implementation prescriptionThe medicament of case 716 wherein L s3 Is- (CH) 2 ) n-C (O) -, where n is 2 or 3.
748. The agent of any one of embodiments 632 to 747, wherein L s3 Ratio of AND L s3 -the amino acid residue to which the bond is closer to the N-terminal amino acid residue.
749. The agent of any one of embodiments 632 to 748, wherein L s3 Bonded to carbon atoms of the peptide backbone.
750. The agent of any one of embodiments 632 to 749, wherein L s3 Bonded to the alpha carbon atom of the amino acid residue.
751. The agent of any one of embodiments 632 to 748, wherein L s3 Bonded to the nitrogen atom of the peptide backbone.
752. The agent of any one of embodiments 632 to 748, wherein L s3 Is bonded to a nitrogen atom of the peptide backbone, wherein the nitrogen atom belongs to an amino group bonded to an alpha carbon atom of an amino acid residue.
753. The agent of any one of embodiments 751-752, wherein the nitrogen atom is attached to L s3 -C (O) -bonding.
754. The agent of any one of embodiments 632-753 wherein the staple-like structure is an optionally substituted-CH 2 -CH＝CH-(CH 2 ) 3 -。
755. The agent of any one of embodiments 632-753 wherein the staple-like structure is-CH 2 -CH＝CH-(CH 2 ) 3 -。
756. The agent of embodiments 754-755, wherein-ch=ch-is E.
757. The agent of embodiments 754-755, wherein-ch=ch-is Z.
758. The agent of any one of embodiments 632-753 wherein the staple-like structure is an optionally substituted-CH 2 -CH＝CH-(CH 2 ) 3 -C(O)-。
759. The agent of any one of embodiments 632-753 wherein the staple-like structure is-CH 2 -CH＝CH-(CH 2 ) 3 -C(O)-。
760. The agent of any one of embodiments 632-753 wherein the staple-like structure is an optionally substituted-CH 2 -CH＝CH-(CH 2 ) 2 -C(O)-。
761. The agent of any one of embodiments 632-753 wherein the staple-like structure is-CH 2 -CH＝CH-(CH 2 ) 2 -C(O)-。
762. The agent of embodiments 758 through 761, wherein-ch=ch-is E.
763. The agent of embodiments 758 through 761, wherein-ch=ch-is Z.
764. The agent of any one of embodiments 632-753 wherein the staple-like structure is optionally substituted- (CH) 2 ) n-, wherein n is 1 to 20.
765. The agent of any one of embodiments 632-753 wherein the staple-like structure is- (CH) 2 ) n-, wherein n is 1 to 20.
766. The agent of any one of embodiments 632-753 wherein the staple-like structure is optionally substituted- (CH) 2 ) n-CO-where n is 1 to 20.
767. The agent of any one of embodiments 632-753 wherein the staple-like structure is- (CH) 2 ) n-C (O) -, wherein n is 1 to 20.
768. The agent of embodiments 764-767 wherein n is 4 to 10.
769. The agent of embodiments 764-767 wherein n is 5-8.
770. The agent of embodiments 764-767 wherein n is 6.
771. The agent of any one of embodiments 754-770, wherein optionally substituted- (CH) 2 ) 3 -or-C (O) -is bonded to an amino acid residue closer to the N-terminus of another amino acid residue bonded to the same staple-like structure.
772. The agent of any one of embodiments 754-771, wherein optionally substituted- (CH) 2 ) 3 -or-C (O) -is bonded to an alpha carbon atom of an amino acid residue.
773. The agent of any one of embodiments 754-771, wherein optionally substituted- (CH) 2 ) 3 -or-C (O) -bonded to the nitrogen atom of the amino acid residue.
774. The agent of any one of embodiments 754-771, wherein optionally substituted- (CH) 2 ) 3 -or-C (O) -is bonded to a nitrogen atom, said nitrogen atom being bonded to an alpha carbon atom of an amino acid residue.
775. The agent of any one of embodiments 754-774, wherein optionally substituted- (CH) 2 ) 3 -or-C (O) -and X 1 And (5) bonding.
776. The agent of embodiment 775, wherein the additional amino acid residue bonded to the staple-like structure is X 3 。
777. The agent of embodiment 775, wherein the additional amino acid residue bonded to the staple-like structure is X 4 。
778. The medicament of any of embodiments 632 to 777, wherein the staple-like structure is- (CH) 2 )m-N(R’)-(CH 2 )n-CH＝CH-(CH 2 ) n '-, where m, n and n' are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and each-CH 2 -independently optionally substituted.
779. The medicament of any of embodiments 632 to 777, wherein the staple-like structure is- (CH) 2 )m-N(R’)-(CH 2 )n-CH＝CH-(CH 2 ) n '-, where m, n, and n' are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
780. The medicament of any of embodiments 632 to 779, wherein the staple-like structure is- (CH) 2 )m-N(R’)-C(O)-O-(CH 2 )n-CH＝CH-(CH 2 ) n '-, where m, n and n' are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and each-CH 2 -independently optionally substituted.
781. The medicament of any of embodiments 632 to 779, wherein the staple-like structure is- (CH) 2 )m-N(R’)-C(O)-O-(CH 2 )n-CH＝CH-(CH 2 ) n' -, where m,n and n' are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.
782. The agent of any one of embodiments 778-781, wherein-ch=ch-is E.
783. The agent of any one of embodiments 778-781, wherein-ch=ch-is Z.
784. The agent of any one of embodiments 632-783, wherein the staple-like structure is- (CH) 2 )m-N(R’)-(CH 2 )n-CH 2 -CH 2 -(CH 2 ) n '-, where m, n and n' are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and each-CH 2 -independently optionally substituted.
785. The medicament of any of embodiments 632 to 784, wherein the staple-like structure is- (CH) 2 )m-N(R’)-(CH 2 )n-CH 2 -CH 2 -(CH 2 ) n '-, where m, n, and n' are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
786. The agent of any one of embodiments 632-785, wherein the staple-like structure is- (CH) 2 )m-N(R’)-C(O)-O-(CH 2 )n-CH 2 -CH 2 -(CH 2 ) n '-, where m, n and n' are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and each-CH 2 -independently optionally substituted.
787. The agent of any one of embodiments 632-786, wherein the staple-like structure is- (CH) 2 )m-N(R’)-C(O)-O-(CH 2 )n-CH 2 -CH 2 -(CH 2 ) n '-, where m, n, and n' are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
788. The agent of any one of embodiments 778-787, wherein- (CH) 2 ) m-is bonded to an amino acid residue closer to the N-terminus of another amino acid residue bonded to the same staple-like structure.
789. The agent of any one of embodiments 778-787, wherein- (CH) 2 ) m-is bonded to an amino acid residue closer to the C-terminus of another amino acid residue bonded to the same staple-like structure.
790. The agent of any one of embodiments 778-789, wherein m is 1.
791. The agent of any one of embodiments 778-789, wherein m is 2.
792. The agent of any one of embodiments 778-791, wherein n is 3.
793. The agent of any one of embodiments 778-792, wherein n' is 3.
794. The agent of any one of embodiments 778-793, wherein R' is-H.
795. The agent of any one of embodiments 778-793, wherein R' is optionally substituted C 1-6 Aliphatic series.
796. The agent of any one of embodiments 778-793, wherein R' is methyl.
797. The agent of any one of embodiments 778-793, wherein R' and L s3 R of bonded amino acid residue a3 And intervening atoms together form an optionally substituted 3-to 10-membered ring having 0 to 5 heteroatoms in addition to the intervening atoms.
798. The agent of any one of embodiments 778-793, wherein R' and L s3 R of bonded amino acid residue a3 And intervening atoms together form a 3 to 10 membered monocyclic ring having 0 to 5 heteroatoms in addition to the intervening atoms.
799. The agent of any one of embodiments 797-798, wherein the ring formed is saturated.
800. The agent of any one of embodiments 797-799, wherein the ring formed is 4-membered.
801. The agent of any one of embodiments 797-800, wherein the ring formed is 5-membered.
802. The agent of any one of embodiments 797-801, wherein the formed ring has no heteroatoms other than the intervening atoms.
803. The agent of any one of embodiments 632-802, wherein the staple-like structure is optionally substituted-CH 2 -N(-CH 2 -**CH 2 -)-C(O)O-(CH 2 ) 3 -CH＝CH-(CH 2 ) 3 -, where-CH 2 -and-CH 2 -bonding to the same amino acid residue.
804. The agent of any one of embodiments 632-802, wherein staple-like structure is CH 2 -N(-CH 2 -**CH 2 -)-C(O)O-(CH 2 ) 3 -CH＝CH-(CH 2 ) 3 -, where-CH 2 -and-CH 2 -bonding to the same amino acid residue.
805. The agent of embodiments 803-804, wherein-ch=ch-is E.
806. The agent of embodiments 803-804, wherein-ch=ch-is Z.
807. The agent of any one of embodiments 632-802, wherein the staple-like structure is optionally substituted-CH 2 -N(-CH 2 -**CH 2 -)-C(O)O-(CH 2 ) 3 -CH 2 -CH 2 -(CH 2 ) 3 -, where-CH 2 -and-CH 2 -bonding to the same amino acid residue.
808. The agent of any one of embodiments 632-802, wherein staple-like structure is CH 2 -N(-CH 2 -**CH 2 -)-C(O)O-(CH 2 ) 3 -CH 2 -CH 2 -(CH 2 ) 3 -, where-CH 2 -and-CH 2 -bonding to the same amino acid residue.
809. The agent of any of embodiments 803 to 808, wherein CH 2 -and-CH 2 -bonding to the same atom.
810. The agent of any one of embodiments 778-809, wherein optionally substituted- (CH) 2 ) m -or-CH 2 -to an amino acid residue closer to the C-terminus of another amino acid residue bonded to the same staple-like structure.
811. The agent of any one of embodiments 778-810, wherein optionally substituted- (CH) 2 ) m -or-CH 2 Alpha carbon atom to amino acid residueSub-bonding.
812. The agent of any one of embodiments 778-811, wherein optionally substituted- (CH) 2 ) m -or-CH 2 -and X 11 And (5) bonding.
813. The agent of embodiment 812, wherein the additional amino acid residue bonded to the staple-like structure is X 4 。
814. The medicament of any of embodiments 632 to 813, wherein the staple-like structure is optionally substituted- (CH) 2 )m-CH＝CH-(CH 2 )n-。
815. The medicament of any of embodiments 632 to 813, wherein the staple-like structure is- (CH) 2 )m-CH＝CH-(CH 2 )n-。
816. The medicament of any of embodiments 632 to 813, wherein the staple-like structure is optionally substituted- (CH) 2 )m-CH 2 -CH 2 -(CH 2 )n-。
817. The medicament of any of embodiments 632 to 813, wherein the staple-like structure is- (CH) 2 )m-CH 2 -CH 2 -(CH 2 )n-。
818. The agent of any one of embodiments 814-817, wherein m is 1.
819. The agent of any one of embodiments 814-817, wherein m is 2.
820. The agent of any one of embodiments 814-817, wherein m is 3.
821. The agent of any one of embodiments 814-817, wherein m is 4.
822. The agent of any one of embodiments 814-817, wherein m is 5.
823. The agent of any one of embodiments 814-817, wherein m is 6.
824. The agent of any one of embodiments 814-817, wherein m is 7.
825. The agent of any one of embodiments 814-817, wherein m is 8.
826. The agent of any one of embodiments 814-825, wherein n is 1.
827. The agent of any one of embodiments 814-825, wherein n is 2.
828. The agent of any one of embodiments 814-825, wherein n is 3.
829. The agent of any one of embodiments 814-825, wherein n is 4.
830. The agent of any one of embodiments 814-825, wherein n is 5.
831. The agent of any one of embodiments 814-825, wherein n is 6.
832. The agent of any one of embodiments 814-825, wherein n is 7.
833. The agent of any one of embodiments 814-825, wherein n is 8.
834. The medicament of any one of embodiments 814 to 833, wherein the staple-like structure is associated with X 4 And X 11 And (5) bonding.
835. The medicament of any of embodiments 632 to 834, wherein the staple-like structure is- (CH) 2 )m-N(R’)-(CH 2 ) n-, wherein m and n are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and each-CH 2 -independently optionally substituted.
836. The medicament of any of embodiments 632 to 835, wherein the staple-like structure is- (CH) 2 )m-N(R’)-(CH 2 ) n-, wherein m and n are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.
837. The medicament of any of embodiments 632 to 836, wherein the staple-like structure is- (CH) 2 )m-N(R’)-C(O)-O-(CH 2 ) n-, wherein m and n are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and each-CH 2 -independently optionally substituted.
838. The agent of any one of embodiments 632 to 837 wherein the staple-like structure is- (CH) 2 )m-N(R’)-C(O)-O-(CH 2 ) n-, wherein m and n are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.
839. The agent of any of embodiments 835 to 838 wherein R' is-H.
840. The agent of any of embodiments 835 to 838 wherein R' is optionally substituted C 1-6 Aliphatic series.
841. The agent of any of embodiments 835 to 838 wherein R' is methyl.
842. The medicament of any of embodiments 632 to 834, wherein the staple-like structure is- (CH) 2 )m-L s2 -(CH 2 ) n-, wherein m and n are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.
843. The agent of embodiment 842, wherein L s2 Is optionally substituted
844. The agent of embodiment 842, wherein L s2 Is that
845. The agent of embodiment 842, wherein L s2 Is optionally substituted
846. The agent of embodiment 842, wherein L s2 Is that
847. The agent of any of embodiments 842-845 wherein the carbon atom is bonded to- (CH) 2 ) m-bonding.
848. The agent of embodiment 842, wherein L s2 is-C (O) -N (R') - (CH) 2 ) n -N (R') -C (O) -, wherein N is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and each-CH 2 -independently optionally substituted.
849. The agent of embodiment 842, wherein L s2 is-C (O) -N (R') - (CH) 2 ) n -N (R') -C (O) -, wherein N is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.
850. The agent of embodiment 842, wherein L s2 is-N (R') - (CH) 2 ) n -N (R') -, wherein N is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and each-CH 2 -independently optionally substituted.
851. The agent of embodiment 842, wherein L s2 is-N (R') - (CH) 2 ) n -N (R') -, wherein N is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.
852. The agent of embodiment 842, wherein L s2 is-C (O) -N (R') - (CH) 2 ) n1 -C(R’) 2 -(CH 2 ) n2 -N (R') -C (O) -, wherein N1 and N2 are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and each-CH 2 -independently optionally substituted.
853. The agent of embodiment 842, wherein L s2 is-C (O) -N (R') - (CH) 2 ) n1 -C(R’) 2 -(CH 2 ) n2 -N (R') -C (O) -, wherein N1 and N2 are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.
854. The agent of embodiment 842, wherein L s2 is-N (R') - (CH) 2 ) n1 -C(R’) 2 -(CH 2 ) n2 -N (R') -, wherein N1 and N2 are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and each-CH 2 -independently optionally substituted.
855. The agent of embodiment 842, wherein L s2 is-N (R') - (CH) 2 ) n1 -C(R’) 2 -(CH 2 ) n2 -N (R') -, wherein N1 and N2 are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.
856. The agent of any one of embodiments 848-855, wherein each R' is independently-H or optionally substituted C 1-6 Aliphatic series.
857. The agent of any of embodiments 848-855, wherein two R' together with an intervening atom form an optionally substituted 3-to 10-membered ring having 0-5 heteroatoms in addition to the intervening atom.
858. The agent of embodiment 857, wherein-C (R') 2 -one R 'and one R' of-N (R ') -or-N (R') C (O) O-together with the intervening atoms form an optionally substituted 3-to 10-membered ring having 0 to 5 heteroatoms in addition to the intervening atoms.
859. The agent of embodiment 857, wherein-C (R') 2 -one R 'and one R' of-N (R ') -or-N (R') C (O) O-together with the intervening atoms form a single ring 3 to 10 membered ring having 0 to 5 heteroatoms in addition to the intervening atoms.
860. The agent of any one of embodiments 857 to 859 wherein the ring formed is saturated.
861. The agent of any one of embodiments 857 to 860 wherein the ring formed is 4-membered.
862. The agent of any one of embodiments 857 to 860 wherein the ring formed is 5 membered.
863. The agent of any one of embodiments 857 to 860 wherein the ring formed is 6 membered.
864. The agent of any one of embodiments 857 to 863 wherein the formed ring has no heteroatoms other than the intervening atoms.
865. The agent of any of embodiments 859 to 864 wherein-C (R') 2 -another R 'and one R' of-N (R ') -or-N (R') C (O) O-together with the intervening atom form an optionally substituted 3-to 10-membered ring having 0 to 5 heteroatoms in addition to the intervening atom.
866. The agent of any one of embodiments 859 to 865 wherein-C (R') 2 -another R ' and-N (R ') -or-N (R ') C (O) O-together with the intervening atoms form a single ring 3 to 10 membered ring having 0 to 5 heteroatoms in addition to the intervening atoms.
867. The agent of any one of embodiments 865 to 866, wherein the ring formed is saturated.
868. The agent of any one of embodiments 865-867 wherein the ring formed is 4-membered.
869. The agent of any one of embodiments 865-867 wherein the ring formed is 5-membered.
870. The agent of any one of embodiments 865-867 wherein the ring formed is 6 membered.
871. The agent of any one of embodiments 865-870, wherein the formed ring has no heteroatoms other than the intervening atoms.
872. The agent of any one of embodiments 852 to 871, wherein n1 is 1.
873. The agent of any one of embodiments 852 to 871, wherein n1 is 2.
874. The agent of any one of embodiments 852 to 873, wherein n2 is 1.
875. The agent of any one of embodiments 852 to 873, wherein n2 is 2.
876. The medicament of any of embodiments 632 to 834, wherein the staple-like structure is-S-CH 2 -Cy-CH 2 S-, each of which is-CH 2 -independently optionally substituted.
877. The agent of embodiment 876, wherein L s2 is-S-CH 2 -Cy-CH 2 -S-。
878. The medicament of any of embodiments 632 to 834, wherein the staple-like structure is-S-CH 2 -Cy-Cy-CH 2 S-, each of which is-CH 2 -independently optionally substituted.
879. The agent of embodiment 878, wherein L s2 is-S-CH 2 -Cy-Cy-CH 2 -S-。
880. The agent of any of embodiments 876-879 wherein-Cy-is optionally substituted phenylene.
881. The agent of embodiment 880 wherein-Cy-is 1, 2-phenylene.
882. The agent of embodiment 880 wherein-Cy-is 1, 3-phenylene.
883. The agent of embodiment 880, wherein-Cy-is 1, 4-phenylene.
884. The agent of any one of embodiments 632-834 wherein the staple-like structure is-S-Cy-S-.
885. The agent of any one of embodiments 632-834, wherein the staple-like structure is-S-Cy-S-.
886. The agent of any one of embodiments 884-885, wherein each-Cy-is optionally substituted phenylene.
887. The agent of any one of embodiments 884-885, wherein each-Cy-is 1, 4-tetrafluorophenylene.
888. The agent of any one of embodiments 632-834, wherein the staple-like structure is-C (O) -Cy-C (O) -.
889. The agent of embodiment 888, wherein-Cy-is an optionally substituted monocyclic or bicyclic 5-to 12-membered ring, wherein each-C (O) -is independently bonded to a nitrogen atom.
890. The agent of any one of embodiments 632 to 834, wherein the staple-like structure is-N (R ') -C (O) -L "-C (O) -N (R') -.
891. The agent of embodiment 890 wherein L "is-Cy-.
892. The agent of embodiment 890 wherein L "is optionally substituted phenylene.
893. The agent of embodiment 890 wherein L "is optionally substituted 1, 3-phenylene.
894. The agent of embodiment 890 wherein L "is an optionally substituted divalent C 1-6 Aliphatic series.
895. The agent of embodiment 890 wherein L "is optionally substituted- (CH) 2 ) n-, wherein n is 1,2, 3, 4, 5, 6, 7, 8, 9 or 10.
896. The agent of embodiment 890 wherein L "is-CH 2 -CH 2 -。
897. The agent of embodiment 890 wherein L "is-C (CH 3 ) 2 -。
898. The agent of embodiment 890 wherein L "is-Cy-.
899. The agent of embodiment 898 wherein each-Cy-is independently optionally substituted phenylene.
900. The agent of embodiment 898 wherein each-Cy-is independently 1, 2-phenylene.
901. The agent of embodiment 898 wherein each-Cy-is independently 1, 3-phenylene.
902. The agent of any one of embodiments 890-901, wherein each R 'in-N (R') -is independently-H or optionally substituted C 1-6 Aliphatic series.
903. The agent of any one of embodiments 890-901, wherein each R 'in-N (R') -is independently-H.
904. The medicament of any of embodiments 632 to 834, wherein the staple-like structure is- (CH) 2 )m-O-CH 2 -L s2 -(CH 2 ) n-, wherein each-CH 2 -independently optionally substituted.
905. The medicament of embodiment 889, wherein the staple-like structure is- (CH) 2 )m-O-CH 2 -L s2 -(CH 2 )n-。
906. The agent of any one of embodiments 889-905, wherein L s2 is-Cy-.
907. The agent of embodiment 906 wherein-Cy-is optionally substituted
908. The agent of embodiment 906, wherein-Cy-is
909. The agent of embodiment 906 wherein-Cy-is optionally substituted
910. The agent of embodiment 906, wherein-Cy-is
911. The agent of any of embodiments 909 to 910 wherein the carbon atom is attached to- (CH) 2 ) n-bonding, the- (CH) 2 ) n-is bonded to an amino acid residue.
912. The agent of any of embodiments 909 through 910 wherein the carbon atom is bonded to-CH 2 -bonding, said-CH 2 -bonding with-O-.
913. The agent of any of embodiments 835 to 912 wherein- (CH) 2 ) m-is bonded to an amino acid residue closer to the N-terminus of another amino acid residue bonded to the same staple-like structure.
914. The agent of any of embodiments 835 to 912 wherein- (CH) 2 ) m-is bonded to an amino acid residue closer to the C-terminus of another amino acid residue bonded to the same staple-like structure.
915. The agent of any one of embodiments 835 to 914, wherein m is 1.
916. The agent of any one of embodiments 835 to 914, wherein m is 2.
917. The agent of any one of embodiments 835 to 914, wherein m is 3.
918. The agent of any one of embodiments 835 to 914, wherein m is 4.
919. The agent of any one of embodiments 835 to 918 wherein n is 1.
920. The agent of any one of embodiments 835 to 918 wherein n is 2.
921. The agent of any one of embodiments 835 to 918 wherein n is 3.
922. The agent of any one of embodiments 835 to 918 wherein n is 4.
923. The agent of any one of embodiments 632-922, wherein the agent comprises an optionally substituted- (CH) 2 ) 2 C(O)NH(CH 2 ) 4 -a staple-like structure.
924. Embodiments 632 to 923The agent of any one of claims, wherein the agent comprises a compound that is- (CH) 2 ) 2 C(O)NH(CH 2 ) 4 -a staple-like structure.
925. The medicament of any of embodiments 632 to 924, wherein the staple-like structure is optionally substituted
926. The medicament of any of embodiments 632 through 925, wherein the staple-like structure is
927. The medicament of any of embodiments 632 through 925, wherein the staple-like structure is
928. The medicament of any of embodiments 632 to 927, wherein the staple-like structure is optionally substituted
929. The medicament of any of embodiments 632 through 928, wherein the staple-like structure is
930. The medicament of any of embodiments 632 through 928, wherein the staple-like structure is
931. The agent of any one of embodiments 923 to 930, wherein optionally substituted- (CH) 2 ) 4 -to an amino acid residue closer to the N-terminus of another amino acid residue bonded to the same staple-like structure.
932. The agent of any one of embodiments 923 to 930, wherein optionally viaSubstituted- (CH) 2 ) 4 -to an amino acid residue closer to the C-terminus of another amino acid residue bonded to the same staple-like structure.
933. The agent of any of embodiments 632-932 wherein the staple-like structure is an optionally substituted-S-CH 2 - (1, 3-phenylene) -CH 2 -S-。
934. The agent of any one of embodiments 632 to 933 wherein the staple-like structure is-S-CH 2 - (1, 3-phenylene) -CH 2 -S-。
935. The staple-like structure of any one of embodiments 835-934, wherein the staple-like structure is aligned with X 3 And X 7 And (5) bonding.
936. The staple-like structure of any one of embodiments 835-934, wherein the staple-like structure is aligned with X 3 And X 10 And (5) bonding.
937. The staple-like structure of any one of embodiments 835-934, wherein the staple-like structure is aligned with X 7 And X 10 And (5) bonding.
938. The staple-like structure of any one of embodiments 835-934, wherein the staple-like structure is aligned with X 7 And X 14 And (5) bonding.
939. The staple-like structure of any one of embodiments 835-934, wherein the staple-like structure is aligned with X 10 And X 14 And (5) bonding.
940. The agent of any of the preceding embodiments, wherein the staple-like structure is 5 to 10 chain atoms in length.
941. The agent of embodiment 940, wherein the length is 5 chain atoms.
942. The agent of embodiment 940, wherein the length is 6 chain atoms.
943. The agent of embodiment 940, wherein the length is 7 chain atoms.
944. The medicament of any one of embodiments 940 to 943, wherein the staple-like structure is an (i, i+2) staple-like structure.
945. The medicament of any one of embodiments 940 to 943, wherein the staple-like structure is an (i, i+3) staple-like structure.
946. The agent of any of the preceding embodiments, wherein the staple-like structure is 7 to 12 chain atoms in length.
947. The agent of embodiment 946 wherein said length is 7 chain atoms.
948. The agent of embodiment 946 wherein said length is 8 chain atoms.
949. The agent of embodiment 946 wherein said length is 9 chain atoms.
950. The medicament of any one of embodiments 946 to 949, wherein the staple-like structure is an (i, i+3) staple-like structure.
951. The agent of any of the preceding embodiments, wherein the staple-like structure is 10 to 25 chain atoms in length.
952. The agent of embodiment 951, wherein the length is 12 chain atoms.
953. The agent of embodiment 951, wherein the length is 13 chain atoms.
954. The agent of embodiment 951, wherein the length is 14 chain atoms.
955. The agent of any one of embodiments 951 to 954, wherein the staple-like structure is an (i, i+7) staple-like structure.
956. The agent of any one of embodiments 630 to 955, wherein the three staple-like structures are within 10 to 20 consecutive amino acid residues.
957. The agent of any one of embodiments 630 to 955, wherein the three staple-like structures are within 14 consecutive amino acid residues.
958. The agent of any one of embodiments 630 to 955, wherein the three staple-like structures are within 11 consecutive amino acid residues.
959. The agent of any one of embodiments 630-958, wherein the first staple-like structure connects two residues at positions i and i+2.
960. The agent of any one of embodiments 630-958, wherein the first staple-like structure connects two residues at positions i and i+3.
961. The agent of any one of embodiments 630-960 wherein the second staple-like structure connects two residues at positions i+3 and i+10.
962. The agent of any one of embodiments 630-961 wherein the third staple-like structure connects two residues at positions i+9 and i+13.
963. The agent of any one of embodiments 630-962, wherein the third staple-like structure connects two residues at positions i+6 and i+9.
964. The agent of any one of embodiments 630-963 wherein the third staple-like structure connects two residues at positions i+6 and i+13.
965. The agent of any one of embodiments 630-964, wherein the peptide comprises a fourth staple-like structure.
966. The agent of any one of embodiments 630-965 wherein the fourth staple structure connects two residues at positions i+2 and i+6.
967. The medicament of any one of embodiments 630 through 966, wherein the first staple-like structure has-L s1 -L s2 -L s3 -a structure.
968. The medicament of any one of embodiments 630 through 967, wherein the second staple-like structure has-L s1 -L s2 -L s3 -a structure.
969. The medicament of any one of embodiments 630 through 968, wherein the third staple-like structure has-L s1 -L s2 -L s3 -a structure.
970. The medicament of any one of embodiments 630 through 969, wherein the fourth staple-like structure has-L s1 -L s2 -L s3 -a structure.
971. The agent of any one of the preceding embodiments, comprising a first staple-like structure comprising (E) -double bonds.
972. The agent of any one of the preceding embodiments, comprising a first staple-like structure comprising a (Z) -double bond.
973. The agent of any one of the preceding embodiments comprising a second staple-like structure comprising (E) -double bonds.
974. The agent of any one of the preceding embodiments comprising a second staple-like structure comprising a (Z) -double bond.
975. The agent of any one of the preceding embodiments comprising a third staple-like structure comprising (E) -double bonds.
976. The agent of any one of the preceding embodiments comprising a third staple-like structure comprising a (Z) -double bond.
977. The agent of any one of the preceding embodiments, wherein X 1 And X is 4 The staple-like structure therebetween has-L s1 -L s2 -L s3 -structure, wherein L s1 、L s2 And L s3 Each independently is a covalent bond or an optionally substituted divalent linear or branched, saturated or partially unsaturated C 1-10 Hydrocarbon chains in which one or more methylene units are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-，-S-Cy-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
978. The agent of any one of the preceding embodiments, wherein X 4 And X is 11 The staple-like structure therebetween has-L s1 -L s2 -L s3 -structure, wherein L s1 、L s2 And L s3 Each independently is a covalent bond or an optionally substituted divalent linear or branched, saturated or partially unsaturated C 1-10 Hydrocarbon chains in which one or more methylene units are optionally and independently replaced by:
-C(R’) 2 -，-Cy-,-O-，-S-，-S-S-，-S-Cy-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
979. The agent of any one of the preceding embodiments, wherein X 10 And X is 14 The staple-like structure therebetween has-L s1 -L s2 -L s3 -structure, wherein L s1 、L s2 And L s3 Each independently is a covalent bond or an optionally substituted divalent linear or branched, saturated or partially unsaturated C 1-10 Hydrocarbon chains in which one or more methylene units are optionally and independently replaced by:
-C(R’) 2 -，-Cy-,-O-，-S-，-S-S-，-S-Cy-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
980. The agent of any one of the preceding embodiments, wherein X 7 And X is 10 The staple-like structure therebetween has-L s1 -L s2 -L s3 -structure, wherein L s1 、L s2 And L s3 Each independently is a covalent bond or an optionally substituted divalent linear or branched, saturated or partially unsaturated C 1-10 Hydrocarbon chains in which one or more methylene units are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-，-S-Cy-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
981. The agent of any one of the preceding embodiments, wherein X 7 And X is 14 The staple-like structure therebetween has-L s1 -L s2 -L s3 -structure, wherein L s1 、L s2 And L s3 Each independently is a covalent bond or an optionally substituted divalent linear or branched, saturated or partially unsaturated C 1-10 Hydrocarbon chains in which one or more methylene units are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-，-S-Cy-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
982. The agent of any one of the preceding embodiments, wherein X 3 And X is 7 The staple-like structure therebetween has-L s1 -L s2 -L s3 -structure, wherein L s1 、L s2 And L s3 Each independently is a covalent bond or an optionally substituted divalent linear or branched, saturated or partially unsaturated C 1-10 Hydrocarbon chains in which one or more methylene units are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-，-S-Cy-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
983. The agent of any one of the preceding embodiments, wherein L s1 Is a covalent bond or an optionally substituted divalent linear or branched, saturated or partially unsaturated C 1-10 Hydrocarbon chains in which one or more methylene units are optionally and independently replaced by-O-.
984. The agent of any one of the preceding embodiments, wherein L s1 Is an optionally substituted divalent linear or branched, saturated or partially unsaturated C 1-10 A hydrocarbon chain.
985. The agent of any one of the preceding embodiments, wherein L s2 is-Cy-.
986. The agent of any one of the preceding embodiments, wherein L s2 Is an optionally substituted triazolylene ring.
987. The agent of any one of embodiments 1 to 984, wherein L s2 is-C (O) -or comprises-C (O) -.
988. The agent of any one of embodiments 1 to 984, wherein L s2 is-C (O) N%R ') -or-C (O) N (R') -.
989. The agent of any one of embodiments 1 to 984 and 988, wherein L s2 is-C (O) NH-.
990. The agent of any one of embodiments 1 to 984 and 988, wherein L s2 is-C (O) N (R ') -wherein R' is C 1-6 Aliphatic series.
991. The agent of any one of embodiments 1 to 984, wherein L s2 is-S-Cy-S-.
992. The agent of any one of embodiments 1 to 984 and 991, wherein L s2 is-S-Cy-S-, wherein-Cy-is an optionally substituted monocyclic or bicyclic arylene ring.
993. The agent of any one of embodiments 1 to 984 and 991 to 992, wherein L s2 is-S-Cy-S-, wherein-Cy-is an optionally substituted phenylene ring.
994. The agent of any one of embodiments 1 to 984 and 991 to 992, wherein L s2 is-S-Cy-S-, wherein-Cy-is an optionally substituted biphenylene ring.
995. The agent of any one of the preceding embodiments, wherein L s3 Is a covalent bond or an optionally substituted divalent linear or branched, saturated or partially unsaturated C 1-10 Hydrocarbon chains in which one or more methylene units are optionally and independently replaced by-O-.
996. The agent of any one of the preceding embodiments, wherein L s3 Is an optionally substituted divalent linear or branched, saturated or partially unsaturated C 1-10 The hydrocarbon chain alternatively comprising optionally substituted divalent linear or branched, saturated or partially unsaturated C 1-10 A hydrocarbon chain.
997. The agent of any one of the preceding embodiments, wherein L s1 Bonded to atoms of the peptide backbone.
998. The agent of any one of the preceding embodiments, wherein L s1 Bonded to the alpha carbon of the amino acid residue.
999. The agent of any one of the preceding embodiments, wherein L s1 Bonded to ring atoms of the ring, whichWherein the ring comprises one or more ring atoms which are atoms of the peptide backbone.
1000. The agent of any one of the preceding embodiments, wherein L s1 Bonded to a ring atom of a ring, wherein the ring comprises an alpha carbon of an amino acid residue.
1001. The agent of any one of the preceding embodiments, wherein L s1 is-C (R') 2 -substitution, wherein-C (R') 2 -one R 'and the R' attached to the backbone together with the intervening atoms thereof form an optionally substituted 3-to 10-membered ring having 0-5 heteroatoms.
1002. The agent of any one of the preceding embodiments, wherein L s1 Wherein one R ' of-N (R ') -and R ' attached to the backbone together with the intervening atoms form an optionally substituted 3-to 10-membered ring having 0-5 heteroatoms.
1003. The agent of any one of the preceding embodiments, wherein R' attached to the backbone is R of an amino acid a2 。
1004. The agent of any of the preceding embodiments, wherein R' and L s1 The atoms of the same residue to which they are bonded are linked.
1005. The agent of any one of the preceding embodiments, wherein L s3 Bonded to atoms of the peptide backbone.
1006. The agent of any one of the preceding embodiments, wherein L s3 Bonded to the alpha carbon of the amino acid residue.
1007. The agent of any one of the preceding embodiments, wherein L s3 Is bonded to a ring atom of a ring, wherein the ring comprises one or more ring atoms, which are atoms of a peptide backbone.
1008. The agent of any one of the preceding embodiments, wherein L s3 Bonded to a ring atom of a ring, wherein the ring comprises an alpha carbon of an amino acid residue.
1009. The agent of any one of the preceding embodiments, wherein L s3 is-C (R') 2 -substitution, wherein-C (R') 2 -one R 'and the R' attached to the backbone together with the intervening atoms thereof form an optionally substituted 3-to 10-membered ring having 0-5 heteroatoms.
1010. The agent of any one of the preceding embodiments, wherein L s3 Wherein one R ' of-N (R ') -and R ' attached to the backbone together with the intervening atoms form an optionally substituted 3-to 10-membered ring having 0-5 heteroatoms. (e.g., R a2 Or another group attached to the alpha carbon is R').
1011. The agent of any one of the preceding embodiments, wherein R' attached to the backbone is R of an amino acid a2 。
1012. The agent of any of the preceding embodiments, wherein R' and L s3 The atoms of the same residue to which they are bonded are linked.
1013. The agent of any of the preceding embodiments, wherein R' is linked to L s3 The same atoms.
1014. The agent of any one of the preceding embodiments, wherein p0 is 1.
1015. The agent of any one of the preceding embodiments, wherein X 0 Is the residue of an amino acid comprising an alkene.
1016. The agent of any one of the preceding embodiments, wherein X 0 Is comprised of-CH=CH 2 Residues of amino acids of (a).
1017. The agent of any one of the preceding embodiments, wherein X 0 Is comprised of-CH=CH 2 And forms a staple-like structure with another amino acid residue by olefin metathesis.
1018. The agent of any one of the preceding embodiments, wherein X 0 Is a residue of an amino acid having the structure of formula A-I, A-II or A-III, wherein R a2 Or R is a3 Comprising olefins
1019. The agent of any one of the preceding embodiments, wherein X 0 Is a residue of an amino acid having the structure of formula A-I, A-II or A-III, wherein R a2 Or R is a3 is-L a -CH＝CH 2 。
1020. The agent of any one of the preceding embodiments, wherein X 0 Is S5 or S6.
1021. The agent of any one of the preceding embodiments, wherein X 0 And X is 4 Stapling.
1022. The agent of any one of embodiments 1 through 1014, wherein X 0 Selected from Gly, sar, and NMebAla.
1023. The agent of any one of embodiments 1-1013 wherein p0 is 0.
1024. The agent of any one of the preceding embodiments, wherein X 2 Selected from Asp, asn, hse, glu, aad, ser, aThr, thr, meAsn, sbMeAsp, rbMeAsp, aMeDAsp, and OAsp.
1025. The agent of any one of the preceding embodiments, wherein X 2 Selected from Asp, asn, hse, glu, aad, ser, and aThr.
1026. The agent of any one of the preceding embodiments, wherein X 2 Comprising a side chain containing an acidic group.
1027. The agent of any one of the preceding embodiments, wherein X 2 Comprising a side chain containing-COOH or a salt form thereof.
1028. The agent of any one of the preceding embodiments, wherein X 2 Is Asp.
1029. The agent of any one of embodiments 1 through 1025, wherein X 2 Comprising a side chain containing a polar group.
1030. The agent of any one of embodiments 1 through 1025 and 1029, wherein X 2 Comprising amide group-containing side chains.
1031. The agent of any one of the preceding embodiments, wherein X 2 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)-。
1032. The agent of embodiment 1031, wherein R a1 is-H.
1033. The agent of any one of embodiments 1031 to 1032, wherein R a3 is-H.
1034. The agent of any one of embodiments 1031 to 1032, wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
1035. The agent of any one of embodiments 1031 to 1034, wherein L a1 Is a covalent bond.
1036. The agent of any one of embodiments 1031 to 1035, wherein L a2 Is a covalent bond.
1037. The agent of any one of embodiments 1031 to 1036, wherein R a2 Is or comprises an acidic or polar group.
1038. The agent of any one of embodiments 1031 to 1037, wherein R a2 is-L' -COOH.
1039. The agent of any one of embodiments 1031 to 1037, wherein R a2 is-L' -Cy-COOH.
1040. The agent of embodiment 1039, wherein-Cy-is optionally substituted phenylene.
1041. The agent of any one of embodiments 1031 to 1037, wherein R a2 is-L '-C (O) N (R') 2 。
1042. The agent of any one of embodiments 1038 to 1041, wherein L "is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1043. The agent of any one of embodiments 1038 to 1041, wherein L "is an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1044. The agent of any one of embodiments 1038 to 1041, wherein L "is a divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1045. The agent of any one of embodiments 1038 to 1042, wherein L "is optionally substituted- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1046. The agent of any one of embodiments 1038 to 1045, wherein L "is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1047. The agent of any one of embodiments 1 through 1025 and 1029 through 1030, wherein X 2 Is Asn.
1048. The agent of any one of embodiments 1 through 1025 and 1029, wherein X 2 Comprising a side chain containing-OH.
1049. The agent of any one of embodiments 1 to 1025, 1029, and 1048, wherein X 2 Is Hse.
1050. The agent of any one of embodiments 1 to 1023, wherein X 2 Is that
Asp, ala, asn, glu, npg, ser, hse, val, S5, S6, acLys, tfeGA, aThr, aad, pro, thr, phe, leu, PL3, gln, isoGlu, meAsn, isoDAsp, rbGlu, sbGlu, aspSH, ile, sbMeAsp, rbMeAsp, aMeDAsp, OAsp,3COOHF,NAsp,3Thi,NGlu,isoDGlu,BztA,Tle,Aib,MePro,Chg,Cha, or DipA.
1051. The agent of any one of the preceding embodiments, wherein X 2 Interact with Gly307 of beta-catenin or corresponding amino acid residue.
1052. The agent of any one of the preceding embodiments, wherein X 2 Interacting with Lys312 of β -catenin or with its corresponding amino acid residue.
1053. The agent of any one of the preceding embodiments, wherein X 3 Selected from the group consisting of
Npg, leu, cha, val, nLeu, ile, phe, cypA, cyLeu, chg, pff, diethA, ala, tyr, trp, ser, aib, phg, dipA, octG, cba, morphNva, and F2PipNva.
1054. The agent of any one of the preceding embodiments, wherein X 3 Comprising one or two hydrophobic side chains.
1055. The agent of any one of the preceding embodiments, wherein X 3 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)-。
1056. The agent of embodiment 1055, wherein X 3 is-N (R) a1 )-C(R a2 )(R a3 )-C(O)-。
1057. The agent of embodiment 1055, wherein X 3 is-NH-C (R) a2 )(R a3 )-C(O)-。
1058. The agent of any one of embodiments 1055-1057, wherein R a2 And R is a3 Independently hydrogen or optionally substituted C 1-10 Aliphatic series.
1059. The agent of any one of embodiments 1055-1057, wherein R a2 And R is a3 One of which is hydrogen and the other is C 1-10 Aliphatic series.
1060. The agent of any one of embodiments 1055-1057, wherein R a2 And R is a3 Taken together with the carbon atoms to which they are attached form an optionally substituted 3-to 8-membered ring having 1 to 3 heteroatoms.
1061. The agent of any one of embodiments 1055-1057, wherein R a2 And R is a3 Taken together with the carbon atom to which it is attached form a 3-to 8-membered cycloalkyl.
1062. The agent of any one of the preceding embodiments, wherein X 3 Is C optionally substituted with one OR more substituents independently selected from-Cy-and-OR 1-10 Alkyl, wherein-Cy-is an optionally substituted divalent 3-to 10-membered monocyclic, bicyclic or polycyclic ring having 0 to 5 heteroatoms;
r is independently C 1-4 An alkyl group; or alternatively
Two C 1-10 The alkyl groups together with the intervening atoms form an optionally substituted 3-to 10-membered ring having 0 to 5 heteroatoms in addition to the intervening atoms.
1063. The agent of any one of the preceding embodiments, wherein X 3 Is C as a side chain 1-10 An alkyl group.
1064. The medicament of any of the preceding embodimentsAn agent, wherein X 3 Are not stapled.
1065. The agent of any one of embodiments 1 to 1052, wherein X 3 Is that
Npg, ile, asp, cha, dipA, chg, leu, B5, cba, S5, ala, glu, allylGly, nLeu, ser, B6, asn, B4, glnR, val, [ Phc ] [ allyl ] Dap, hse, [ Bn ] [ allyl ] Dap, IMeK, R5, phe, cypA, cyLeu, pff, diethA, tyr, trp, aib, phg, octG, morpnva, F2Pipnva, [ Piv ] [ allyl ] Dap, [ CyCO ] [ allyl ] Dap, lys, or S3.
1066. The agent of any one of embodiments 1 to 1052, wherein X 3 Is Npg.
1067. The agent of any one of embodiments 1 to 1052, wherein X 3 Is Ile.
1068. The agent of any one of embodiments 1 to 1052, wherein X 3 Is Cha.
1069. The agent of any one of embodiments 1 to 1052, wherein X 3 Is DipA.
1070. The agent of any one of embodiments 1 to 1052, wherein X 3 Is Chg.
1071. The agent of any one of embodiments 1 to 1052, wherein X 3 Is Leu.
1072. The agent of any one of embodiments 1 to 1052, wherein X 3 Is B5.
1073. The agent of any one of embodiments 1 to 1052, wherein X 3 Is Asp.
1074. The agent of any one of embodiments 1 to 1052, wherein X 3 Is Cba.
1075. The agent of any one of embodiments 1 to 1052, wherein X 3 Is S5.
1076. The agent of any one of embodiments 1 to 1052, wherein X 3 Is Ala.
1077. The agent of any one of the preceding embodiments, wherein X 3 Interacts with Tyr306 of β -catenin or with its corresponding amino acid residue.
1078. The foregoing embodimentThe agent of any one of the schemes, wherein X 5 Selected from Asp, glu, asn, hse, aThr, aad, ser, thr, meAsn, sbMeAsp, and RbMeAsp.
1079. The agent of any one of the preceding embodiments, wherein X 5 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)-。
1080. The agent of embodiment 1079, wherein R a1 is-H.
1081. The agent of any one of embodiments 1079-1080, wherein R a3 is-H.
1082. The agent of any one of embodiments 1079-1080, wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
1083. The agent of any one of embodiments 1079-1082, wherein L a1 Is a covalent bond.
1084. The agent of any one of embodiments 1079-1083, wherein L a2 Is a covalent bond.
1085. The agent of any one of embodiments 1079-1084, wherein R a2 Is or comprises an acidic or polar group.
1086. The agent of any one of embodiments 1079-1085, wherein R a2 is-L' -COOH.
1087. The agent of any one of embodiments 1079-1085, wherein R a2 is-L' -Cy-COOH.
1088. The agent of embodiment 1087, wherein-Cy-is optionally substituted phenylene.
1089. The agent of any one of embodiments 1079-1085, wherein R a2 is-L '-C (O) N (R') 2 。
1090. The agent of any one of embodiments 1086-1089, wherein L "is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1091. The agent of any one of embodiments 1086-1089, wherein L "is an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1092. The agent of any one of embodiments 1086-1089, wherein L "is a divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1093. The agent of any one of embodiments 1086-1090, wherein L "is optionally substituted- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1094. The agent of any one of embodiments 1086-1090, wherein L "is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1095. The agent of any one of the preceding embodiments, wherein X 5 Comprising a side chain containing an acidic group.
1096. The agent of any one of the preceding embodiments, wherein X 5 Comprising a side chain containing-COOH or a salt form thereof.
1097. The agent of any one of the preceding embodiments, wherein X 5 Is Asp.
1098. The agent of any one of embodiments 1 to 1078, wherein X 5 Comprising a side chain containing a polar group.
1099. The agent of any one of embodiments 1 to 1078 and 1098, wherein X 5 Comprising a side chain containing-OH.
1100. The agent of any one of embodiments 1 to 1078 and 1098, wherein X 5 Comprising amide group-containing side chains.
1101. The agent of any one of embodiments 1 to 1077, wherein X 5 Selected from the group consisting of
3COOHF, tfeGA, asp, gln, [ CH2CMe2CO2H ] TriAzDap, thr, glu,2OH3COOHF, 4COOHF,2COOHF, his, tyr,5F3Me2COOHF,4F3Me2COOHF,5F3Me3COOHF,4F3Me3COOHF,3F2COOHF, val, ser, trp, asn, ala, arg, dGlu, aThr, hTyr,3cbmf, leu, phe, lys, and Ile.
1102. The agent of any one of embodiments 1 to 1077, wherein X 5 Asp, B5,3COOHF, glu, asn, npg, hse, aThr, aad, ser, thr, meAsn, aspSH, sbMeAsp or RbMeAsp.
1103. The agent of any one of embodiments 1 to 1077, wherein X 5 Is B5.
1104. The agent of any one of embodiments 1 to 1077, wherein X 5 Is 3COOHF.
1105. The agent of any one of embodiments 1 to 1077, wherein X 5 Is Glu.
1106. The agent of any one of the preceding embodiments, wherein X 5 Interact with Trp383 of beta-catenin or with its corresponding amino acid residue.
1107. The agent of any one of the preceding embodiments, wherein X 5 Interact with Arg386 of beta-catenin or with its corresponding amino acid residue.
1108. The agent of any one of the preceding embodiments, wherein X 5 Interact with Asn387 of β -catenin or with its corresponding amino acid residue.
1109. The agent of any one of the preceding embodiments, wherein X 6 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)-。
1110. The agent of embodiment 1109, wherein R a1 is-H.
1111. The agent of any one of embodiments 1109 to 1110, wherein R a3 is-H.
1112. The agent of any one of embodiments 1109 to 1110, wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
1113. The agent of any one of embodiments 1109 to 1112, wherein L a1 Is a covalent bond.
1114. The agent of any one of embodiments 1109 to 1113, whichMiddle L a2 Is a covalent bond.
1115. The agent of any one of embodiments 1109 to 1114, wherein R a2 Is or comprises an acidic or polar group.
1116. The agent of any one of embodiments 1109 to 1115, wherein R a2 is-L' -COOH.
1117. The agent of any one of embodiments 1109 to 1115, wherein R a2 is-L' -Cy-COOH.
1118. The agent of embodiment 1117, wherein-Cy-is optionally substituted phenylene.
1119. The agent of any one of embodiments 1109 to 1115, wherein R a2 is-L '-C (O) N (R') 2 。
1120. The agent of any one of embodiments 1116 to 1119, wherein L "is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1121. The agent of any one of embodiments 1116 to 1119, wherein L "is an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1122. The agent of any of embodiments 1116 to 1119, wherein L "is a divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1123. The agent of any one of embodiments 1116 to 1120 wherein L "is optionally substituted- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1124. The agent of any one of embodiments 1116 to 1123, wherein L "is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1125. The agent of any one of embodiments 1116 to 1122 wherein the methylene unit is replaced with-N (R') -.
1126. The agent of embodiment 1125 wherein R' is-H.
1127. The agent of embodiment 1125 wherein R' is optionally substituted C 1-6 An alkyl group.
1128. The agent of any one of the preceding embodiments, wherein X 6 Comprising side chains containing acidic or polar groups.
1129. The agent of any one of the preceding embodiments, wherein X 6 Comprising a side chain containing an acidic group.
1130. The agent of any one of the preceding embodiments, wherein X 6 Comprising a side chain containing-COOH or a salt form thereof.
1131. The agent of any one of the preceding embodiments, wherein X 6 Is 3COOHF.
1132. The agent of any one of embodiments 1 to 1130, wherein X 6 Is TfeGA.
1133. The agent of any one of embodiments 1 to 1130, wherein X 6 Is Asp.
1134. The agent of any one of embodiments 1 to 1130, wherein X 6 Is [ CH2CMe2CO2H ]]TriAzDap。
1135. The agent of any one of embodiments 1 to 1109, wherein X 6 Comprising a side chain containing a polar group.
1136. The agent of any one of embodiments 1 to 1109 and 1135, wherein X 6 Comprising a side chain containing-OH.
1137. The agent of any one of embodiments 1 to 1109 and 1135, wherein X 6 Comprising amide group-containing side chains.
1138. The agent of any one of embodiments 1 to 1109, 1135 and 1137, wherein X 6 Is Gln.
1139. The agent of any one of the preceding embodiments, wherein X 6 Interacts with Tyr306 of β -catenin or with its corresponding amino acid residue.
1140. The foregoing embodimentThe agent according to any one of the above, wherein X 6 Interacting with Lys345 of β -catenin or with its corresponding amino acid residue.
1141. The agent of any one of the preceding embodiments, wherein X 7 Is a hydrophobic amino acid residue.
1142. The agent of any one of the preceding embodiments, wherein X 7 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)-。
1143. The agent of any one of the preceding embodiments, wherein X 7 is-N (R) a1 )-C(R a2 )(R a3 )-C(O)-。
1144. The agent of any one of the preceding embodiments, wherein X 7 is-NH-C (R) a2 )(R a3 )-C(O)-。
1145. The agent of any one of embodiments 1142 to 1144, wherein R a2 And R is a3 Independently hydrogen or optionally substituted C 1-10 Aliphatic series.
1146. The agent of any one of embodiments 1142 to 1144, wherein R a2 And R is a3 One of which is hydrogen and the other is C 1-10 Aliphatic series.
1147. The agent of any one of embodiments 1142 to 1144, wherein R a2 And R is a3 Taken together with the carbon atoms to which they are attached form an optionally substituted 3-to 8-membered ring having 1 to 3 heteroatoms.
1148. The agent of any one of embodiments 1142 to 1144, wherein R a2 And R is a3 Taken together with the carbon atom to which it is attached form a 3-to 8-membered cycloalkyl.
1149. The agent of any one of the preceding embodiments, wherein X 7 Selected from the group consisting of
Aib, ala, morphGLn, gln, ser, iPrLys, nLeu, cha, hse, npg, val, cyLeu, thr, phe, acp, asn, daMeS, aMeDF, leu, cpg, cbg, me2Gln, met2O, acLys, his, aMeL, daMeL, aMeV, aMeS, and aMeF.
1150. Any of the foregoing embodimentsThe agent of item, wherein X 7 Selected from Aib, ala, morphGLn, gln, ser, iPrLys, nLeu, cha, hse, npg, val, and CyLeu.
1151. The agent of any one of the preceding embodiments, wherein X 7 Selected from Aib, ala, morphGLn, gln, ser, iPrLys, nLeu, cha, and Hse.
1152. The agent of any one of the preceding embodiments, wherein X 7 Is Aib.
1153. The agent of any one of embodiments 1 to 1140, wherein X 7 Is Ala.
1154. The agent of any one of embodiments 1 to 1140, wherein X 7 Is CyLeu.
1155. The agent of any one of embodiments 1 to 1140, wherein X 7 Is Phe.
1156. The agent of any one of embodiments 1 to 1140, wherein X 7 Is nLeu.
1157. The agent of any one of embodiments 1 to 1140, wherein X 7 Is Val.
1158. The agent of any one of the preceding embodiments, wherein X 8 Is a hydrophobic amino acid residue.
1159. The agent of any one of the preceding embodiments, wherein X 8 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)-。
1160. The agent of any one of the preceding embodiments, wherein X 8 is-N (R) a1 )-C(R a2 )(R a3 )-C(O)-。
1161. The agent of any one of the preceding embodiments, wherein X 8 is-NH-C (R) a2 )(R a3 )-C(O)-。
1162. The agent of any one of embodiments 1159-1161, wherein R a2 And R is a3 Independently hydrogen or optionally substituted C 1-10 Aliphatic series.
1163. The agent of any one of embodiments 1159-1161, wherein R a2 And R is a3 One of which is hydrogen, andand the other is C 1-10 Aliphatic series.
1164. The agent of any one of embodiments 1159-1161, wherein R a2 And R is a3 Taken together with the carbon atoms to which they are attached form an optionally substituted 3-to 8-membered ring having 1 to 3 heteroatoms.
1165. The agent of any one of embodiments 1159-1161, wherein R a2 And R is a3 Taken together with the carbon atom to which it is attached form a 3-to 8-membered cycloalkyl.
1166. The agent of any one of the preceding embodiments, wherein X 8 Selected from Ala, aib, cpg, val, leu, gln, lys, asp, glu, aad, nLeu, cba, ser, thr, aThr, morphGLn, phe, hPHE, hTyr, and AcLys.
1167. The agent of any one of embodiments 1 through 1157, wherein X 8 Ala, aib, phc, asp,3COOHF, aThr, gly, ser, nLeu, thr, cpg, val, leu, gln, lys, glu, aad, cba, morphGLn, hPr, hTyr, or AeLys.
1168. The agent of any one of the preceding embodiments, wherein X 8 Is Ala.
1169. The agent of any one of embodiments 1 through 1157, wherein X 8 Is Aib.
1170. The agent of any one of embodiments 1 through 1157, wherein X 8 Is Phe.
1171. The agent of any one of embodiments 1 through 1157, wherein X 8 Is Asp.
1172. The agent of any one of embodiments 1 through 1157, wherein X 8 Is 3COOHF.
1173. The agent of any one of the preceding embodiments, wherein X 8 Interact with Trp383 of beta-catenin or with its corresponding amino acid residue.
1174. The agent of any one of the preceding embodiments, wherein X 9 Selected from the group consisting of
Phe,3COOHF,2NapA, nLeu, tyr,3Thi,4FF,4ClF,4BrF,3FF,3ClF,3BrF,2FF,3OMeF,4CNF,3CNF,4MeF,3MeF, aic, rbipPrF, sbipPrF, rbipPrDF, rbMeXyl A, rbMeXyl DA, cba, cypA, bztA,1NapA, trp, leu, ile, ser,2Thi, chg, hse,4TriA,3F3MeF, thr, his, val, asn, gln,2Cpg, sbMeXyl A, and SbMeXyl DA.
1175. The agent of any one of the preceding embodiments, wherein X 9 Comprising a side chain which is or comprises an optionally substituted aromatic group.
1176. The agent of any one of the preceding embodiments, wherein X 9 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)-。
1177. The agent of embodiment 1176 wherein R a1 is-H.
1178. The agent of any one of embodiments 1176-1177, wherein R a3 is-H.
1179. The agent of any one of embodiments 1176-1177, wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
1180. The agent of any one of embodiments 1176-1179, wherein L a1 Is a covalent bond.
1181. The agent of any one of embodiments 1176-1180, wherein R a2 is-L a -R, wherein R is or comprises an aromatic group.
1182. The agent of embodiment 1181 wherein R is an optionally substituted 6-to 10-membered aryl.
1183. The agent of embodiment 1181 wherein R is optionally substituted phenyl.
1184. The agent of embodiment 1181 wherein R is phenyl.
1185. The agent of embodiment 1181, wherein R is optionally substituted naphthyl.
1186. The agent of embodiment 1181 wherein R is naphthyl.
1187. The agent of embodiment 1181 wherein R is an optionally substituted 5-membered heteroaryl having 1 to 4 heteroatoms.
1188. The agent of embodiment 1181 wherein R is an optionally substituted 6-membered heteroaryl having 1 to 4 heteroatoms.
1189. The agent of embodiment 1181 wherein R is an optionally substituted 9-membered bicyclic heteroaryl having 1 to 4 heteroatoms.
1190. The agent of embodiment 1181 wherein R is an optionally substituted 10 membered bicyclic heteroaryl having 1 to 4 heteroatoms.
1191. The agent of any one of embodiments 1187-1190, wherein the heteroatom is nitrogen.
1192. The agent of any one of embodiments 1187-1191 wherein the heteroatom is oxygen.
1193. The agent of any one of embodiments 1187-1192, wherein the heteroatom is sulfur.
1194. The agent of any one of embodiments 1187-1190, wherein the heteroaryl has only one heteroatom.
1195. The agent of embodiment 1194, wherein the heteroatom is nitrogen.
1196. The agent of embodiment 1194, wherein the heteroatom is oxygen.
1197. The agent of embodiment 1194, wherein the heteroatom is sulfur.
1198. The agent of any one of embodiments 1181 to 1197, wherein L a Is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1199. The agent of any one of embodiments 1181 to 1197, wherein L a Is an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1200. The agent of any one of embodiments 1181 to 1197, wherein L a Is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independentlyThe site is-O-, -S-, -and-N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-, substitution.
1201. The agent of embodiment 1198, wherein L a Is optionally substituted- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1202. The agent of embodiment 1198, wherein L a Is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1203. The agent of embodiment 1198, wherein L a is-CH 2 -。
1204. The agent of any one of the preceding embodiments, wherein X 9 Comprising a side chain which is OR comprises an optionally substituted aromatic group, wherein each optional substituent of the aromatic group is independently selected from the group consisting of halogen, -OR, -R, -C (O) OR, and-CN, wherein each R is independently-H, C 1-4 Alkyl or haloalkyl.
1205. The agent of any one of the preceding embodiments, wherein X 9 Comprising a side chain which is OR comprises an optionally substituted aromatic group, wherein each optional substituent of the aromatic group is independently selected from the group consisting of halogen, -OR, -R, -C (O) OH, and-CN, wherein each R is independently C 1-4 Alkyl or haloalkyl.
1206. The agent of any one of the preceding embodiments, wherein X 9 Comprising a side chain which is OR comprises an optionally substituted aromatic group, wherein each optional substituent of the aromatic group is independently selected from the group consisting of halogen, -OR, -R, -C (O) OH, and-CN, wherein each R is independently C 1-2 Alkyl or haloalkyl.
1207. The agent of any one of the preceding embodiments, wherein X 9 Comprising a side chain which is OR comprises an optionally substituted aromatic group, wherein each optional substituent of the aromatic group is independently selected from halogen, -OR, -R, -C (O) OH and-CN, wherein each R is independently Optionally methyl optionally substituted with one or more halogens.
1208. The agent of any one of the preceding embodiments, wherein X 9 Comprising a side chain that is OR comprises an optionally substituted aromatic group, wherein each optional substituent of the aromatic group is independently selected from the group consisting of halogen, -OR, -R, -C (O) OH, and-CN, wherein each R is independently methyl optionally substituted with one OR more F.
1209. The agent of any one of the preceding embodiments, wherein X 9 Comprising a side chain which is or comprises an optionally substituted aromatic group, wherein each optional substituent of the aromatic group is independently selected from-F, -Cl, -Br, -OCH 3 、-CH 3 、-CF 3 -C (O) OH and-CN.
1210. The agent of any one of the preceding embodiments, wherein X 9 Comprising a side chain which is or comprises an unsubstituted aromatic group.
1211. The agent of any one of embodiments 1 to 1173, wherein X 9 Is that
AA9, phe, ala, lys,3COOHF, aib,2NapA, nLeu,2Thi, tyr,3Thi,4FF,4ClF,4BrF,3FF,3ClF,3BrF,2FF,3OMeF,4CNF,3CNF,4MeF,3MeF, aic, rbipPrF, sbipPrF, rbipPrDF, rbMeXyl A, rbMeXyl DA, cba, cypA, bztA,1NapA, trp, leu, ile, scrChg, hsc,4TriA,3F3McF, thr, his, val, asn, gln,2Cpg, sbMcXyl A, or SbMcXyl DA.
1212. The agent of any one of embodiments 1 to 1173, wherein X 9 Is Phe.
1213. The agent of any one of embodiments 1 to 1173, wherein X 9 Is Ala.
1214. The agent of any one of embodiments 1 to 1173, wherein X 9 Is Lys.
1215. The agent of any one of embodiments 1 to 1173, wherein X 9 Is 3COOHF.
1216The agent of any one of embodiments 1 to 1173, wherein X 9 Is Aib.
1217. The agent of any one of the preceding embodiments, wherein X 9 Interacting with Lys345 of β -catenin or with its corresponding amino acid residue.
1218. The agent of any one of the preceding embodiments, wherein X 9 Interact with Trp383 of beta-catenin or with its corresponding amino acid residue.
1219. The agent of any one of the preceding embodiments, wherein X 10 Are not stapled.
1220. The agent of any one of the preceding embodiments, wherein X 10 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)-。
1221. The agent of embodiment 1220, wherein R a1 is-H.
1222. The agent of any one of embodiments 1220 to 1221, wherein R a3 is-H.
1223. The agent of any one of embodiments 1220 to 1221, wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
1224. The agent of any one of embodiments 1220 to 1223, wherein L a1 Is a covalent bond.
1225. The agent of any one of embodiments 1220-1224, wherein L a2 Is a covalent bond.
1226. The agent of any one of embodiments 1220 to 1225, wherein R a2 is-L' -R.
1227. The agent of any one of embodiments 1220 to 1225, wherein R a2 is-L' -Cy-R.
1228. The agent of any one of embodiments 1226-1227, wherein R is hydrogen or optionally substituted C 1-10 Aliphatic series.
1229. The agent of any one of embodiments 1226-1227, wherein R is optionally substituted C 1-10 Aliphatic series.
1230. Embodiment 1226 to1227 wherein R is C 1-10 Aliphatic series.
1231. The agent of any one of embodiments 1226-1227, wherein R is C 1-10 An alkyl group.
1232. The agent of any one of embodiments 1226-1227, wherein R is optionally substituted phenyl.
1233. The agent of any one of embodiments 1220-1225, R a2 is-L '-C (O) N (R') 2 。
1234. The agent of any one of embodiments 1220-1225, R a2 is-L' -OH.
1235. The agent of any of embodiments 1220-1234, wherein L "is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1236. The agent of any of embodiments 1220-1234, wherein L "is an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1237. The agent of any of embodiments 1220-1234, wherein L "is a divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1238. The agent of any of embodiments 1220-1234, wherein L "is optionally substituted- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1239. The agent of any of embodiments 1220-1234, wherein L "is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1240. The agent of any one of embodiments 1 to 1218, wherein X 10 Is that
Lys, phe, triAzLys, glnR, leu, pyrS2, aib, ala, sAla, asnR, hGLnR, dOrn, pyrS1, dLys, dDab, [ mPyr ] Cys, pyrS3, iPrLys, [ mXyl ] Cys, triAzOrn,1MeK, [ C3] Cys, [ IsoE ] Cys, DGlnR, orn, [ mPyr ] hCys, [ Red ] Cys, [ C3] hCys,4PipA, sCH2S, [ FBB ] Cys, [ pXyl ] hCys, [33Oxe ] Cys, [ Red ] hCys, [ IsoE ] hCys, [13Ac ] hCys, [ m5Meb ] Cys, [ m5Meb ] hCys, ginS3APyr, asnMeEDA, asnR APyr, [ m5Pyr ] Cys, [ m5OMeb ] Cys, [4FB ] Cys, [ oXyl ] Cys, NMeOrn, [2_6-naph ] Cys, [3_3-biPh ] Cys, [ mXyl ] hCys, [3_3-biPh ] hCys, [2_6-naph ] hCys, [33Oxe ] hCys, [13Ac ] Cys, glnR3APyr, asnS3APyr, [ IsoE ] hCys Ox, or [ m5Pyr ] hCys.
1241. The agent of any one of embodiments 1 to 1218, wherein X 10 Is Lys.
1242. The agent of any one of embodiments 1 to 1218, wherein X 10 Is Phe.
1243. The agent of any one of embodiments 1 to 1218, wherein X 10 Is TriAzLys.
1244. The agent of any one of embodiments 1 to 1218, wherein X 10 Is GlnR.
1245. The agent of any one of embodiments 1 to 1218, wherein X 10 Is Leu.
1246. The agent of any one of embodiments 1 to 1218, wherein X 10 Is pyrS2.
1247. The agent of any one of embodiments 1 to 1218, wherein X 10 Is Aib.
1248. The agent of any one of embodiments 1 to 1218, wherein X 10 Is Ala.
1249. The agent of any one of embodiments 1 to 1218, wherein X 10 Is Leu.
1250. The agent of any one of the preceding embodiments, wherein X 12 Selected from the group consisting of
3Thi,2F3MeF, phe, nLeu,2COOHF, cypA,2ClF, ala, abu, leu, hLeu, npg, cpa, nva, cba, chA,2FurA,2OMeF,2MeF,2BrF,2CNF,2NO2F,2PyrA,3PyrA,4PyrA, his,1NapA, val, ile, chg, diethA, hnLeu, octG,2Thi, and 2cbmF.
1251. The agent of any one of the preceding embodiments, wherein X 12 Comprising a side chain which is or comprises an optionally substituted aromatic group.
1252. The agent of any one of the preceding embodiments, wherein X 12 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)-。
1253. The agent of embodiment 1252, wherein R a1 is-H.
1254. The agent of any one of embodiments 1252 through 1253, wherein R a3 is-H.
1255. The agent of any one of embodiments 1252 through 1253, wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
1256. The agent of any one of embodiments 1252 through 1255, wherein L a1 Is a covalent bond.
1257. The agent of any one of embodiments 1252 through 1256, wherein R a2 is-L a -R, wherein R is or comprises an aromatic group.
1258. The agent of embodiment 1257 wherein R is an optionally substituted 6-to 10-membered aryl.
1259. The agent of embodiment 1257 wherein R is optionally substituted phenyl.
1260. The agent of embodiment 1257 wherein R is phenyl.
1261. The agent of embodiment 1257 wherein R is optionally substituted naphthyl.
1262. The agent of embodiment 1257 wherein R is naphthyl.
1263. The agent of embodiment 1257 wherein R is an optionally substituted 5-membered heteroaryl having 1 to 4 heteroatoms.
1264. The agent of embodiment 1257 wherein R is an optionally substituted 6-membered heteroaryl having 1 to 4 heteroatoms.
1265. The agent of embodiment 1257 wherein R is an optionally substituted 9-membered bicyclic heteroaryl having 1 to 4 heteroatoms.
1266. The agent of embodiment 1257 wherein R is an optionally substituted 10 membered bicyclic heteroaryl having 1 to 4 heteroatoms.
1267. The agent of any one of embodiments 1263-1266, wherein the heteroatom is nitrogen.
1268. The agent of any one of embodiments 1263-1267, wherein the heteroatom is oxygen.
1269. The agent of any one of embodiments 1263-1268, wherein the heteroatom is sulfur.
1270. The agent of any one of embodiments 1263-1266, wherein the heteroaryl has only one heteroatom.
1271. The agent of embodiment 1270, wherein said heteroatom is nitrogen.
1272. The agent of embodiment 1270, wherein said heteroatom is oxygen.
1273. The agent of embodiment 1270, wherein said heteroatom is sulfur.
1274. The agent of any one of embodiments 1257 through 1273, wherein L a Is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1275. The agent of any one of embodiments 1257 through 1273, wherein L a Is an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1276. The agent of any one of embodiments 1257 through 1273, wherein L a Is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1277. The agent of embodiment 1274, wherein L a Is optionally substituted-(CH 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1278. The agent of embodiment 1274, wherein L a Is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1279. The agent of embodiment 1274, wherein L a is-CH 2 -。
1280. The agent of any one of the preceding embodiments, wherein X 12 Comprising a side chain which is OR comprises an optionally substituted aromatic group, wherein each optional substituent of the aromatic group is independently selected from halogen, -OR, -R, -C (O) OR, -C (O) N (R) 2 -CN and-NO 2 Wherein each R is independently-H, C 1-4 Alkyl or haloalkyl.
1281. The agent of any one of the preceding embodiments, wherein X 12 Comprising a side chain which is OR comprises an optionally substituted aromatic group, wherein each optional substituent of the aromatic group is independently selected from halogen, -OR, -R, -C (O) OH, -C (O) NH 2 -CN and-NO 2 Wherein each R is independently C 1-4 Alkyl or haloalkyl.
1282. The agent of any one of the preceding embodiments, wherein X 12 Comprising a side chain which is OR comprises an optionally substituted aromatic group, wherein each optional substituent of the aromatic group is independently selected from halogen, -OR, -R, -C (O) OH, -C (O) NH 2 -CN and-NO 2 Wherein each R is independently C 1-2 Alkyl or haloalkyl.
1283. The agent of any one of the preceding embodiments, wherein X 12 Comprising a side chain which is OR comprises an optionally substituted aromatic group, wherein each optional substituent of the aromatic group is independently selected from halogen, -OR, -R, -C (O) OH, -C (O) NH 2 -CN and-NO 2 Wherein each R is independently optionally substituted with one or moreA halogen substituted methyl group.
1284. The agent of any one of the preceding embodiments, wherein X 12 Comprising a side chain which is OR comprises an optionally substituted aromatic group, wherein each optional substituent of the aromatic group is independently selected from halogen, -OR, -R, -C (O) OH, -C (O) NH 2 -CN and-NO 2 Wherein each R is independently methyl optionally substituted with one or more F.
1285. The agent of any one of the preceding embodiments, wherein X 12 Comprising a side chain which is or comprises an optionally substituted aromatic group, wherein each optional substituent of the aromatic group is independently selected from-Br, -OCH 3 、-CH 3 、-CF 3 、-C(O)OH、-C(O)NH 2 -CN or-NO 2 。
1286. The agent of any one of the preceding embodiments, wherein X 12 Comprising a side chain which is an optionally substituted aromatic group optionally substituted in the 2 '-position or comprises an optionally substituted aromatic group optionally substituted in the 2' -position.
1287. The agent of any one of the preceding embodiments, wherein X 12 Comprising a side chain which is or comprises an unsubstituted aromatic group.
1288. The agent of any one of embodiments 1251 through 1287, wherein the aromatic group is a 5-membered heteroaryl.
1289. The agent of any one of the preceding embodiments, wherein X 12 Is 3Thi.
1290. The agent of any one of embodiments 1251 through 1287, wherein the aromatic group is phenyl.
1291. The agent of any one of embodiments 1290, wherein X 12 Is 2F3MeF.
1292. The agent of any one of embodiments 1290, wherein X 12 Is Phe.
1293The agent of any one of embodiments 1290, wherein X 12 Is Phe, wherein the phenyl group is 2' -substituted.
1294. The agent of any one of embodiments 1290, wherein X 12 Is 2F3MeF,2COOHF,2ClF,2OMeF,2MeF,2BrF,2CNF,2NO2F, or 2cbmF.
1295. The agent of any of embodiments 1 through 1249, wherein X 12 Is that
3Thi, phe,2F3MeF, pyrS2,2ClF, hnLeu, bztA,2Thi,2MeF,2FF,34ClF, lys, nLeu,2COOHF,2PhF, hCbA, hCypA, hCHA, cypA, hpe, dipA, hepG, dap7Abu, hhLeu, hhSer, hexG, [2IAPAc ]2NH2F, ala, abu, leu, hleu, npg, cpa, pyrS1, [ Bnc ]2NH2F, [ Phc ]2NH2F, [ Biph ]2NH2F, [3PyAc ]2NH2F, nva, cba, chA,2 OMA, 2F, 2CNF,2 NOF, 2 yrA,3PyrA,4, pyrA, prA, pyrA, 2F, pmA, pyhPa, prA, P2F, pmGac, or [ PmC ]2NH2F, pcOc, pcF.
1296. The agent of any of embodiments 1 through 1249, wherein X 12 Is 3Thi.
1297. The agent of any of embodiments 1 through 1249, wherein X 12 Is Phe.
1298. The agent of any of embodiments 1 through 1249, wherein X 12 Is 2F3MeF.
1299. The agent of any of embodiments 1 through 1249, wherein X 12 Is pyrS2.
1300. The agent of any of embodiments 1 through 1249, wherein X 12 Is 2ClF.
1301. The agent of any of embodiments 1 through 1249, wherein X 12 Is hnLeu.
1302. The agent of any of embodiments 1 through 1249, wherein X 12 Is BztA.
1303. The agent of any of embodiments 1 through 1249, wherein X 12 Is 2Thi.
1304. The agent of any of embodiments 1 through 1249, wherein X 12 Is 2MeF.
1305. The agent of any of embodiments 1 through 1249, wherein X 12 Is 2FF.
1306. The agent of any of embodiments 1 through 1249, wherein X 12 34ClF.
1307. The agent of any one of the preceding embodiments, wherein X 12 Interact with Trp383 of beta-catenin or with its corresponding amino acid residue.
1308. The agent of any one of the preceding embodiments, wherein X 12 Interact with Asn415 of β -catenin or with its corresponding amino acid residue.
1309. The agent of any one of the preceding embodiments, wherein X 13 Comprises optionally substituted aromatic groups.
1310. The agent of any one of the preceding embodiments, wherein X 13 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)-。
1311. The agent of embodiment 1310, wherein R a1 is-H.
1312. The agent of any one of embodiments 1310 to 1311, wherein R a3 is-H.
1313. The agent of any one of embodiments 1310 to 1311, wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
1314. The agent of any one of embodiments 1310 to 1313, wherein L a1 Is a covalent bond.
1315. The agent of any one of embodiments 1310 to 1314, wherein R a2 is-L a -R, wherein R is or comprises an aromatic group.
1316. The agent of embodiment 1315, wherein R is an optionally substituted 6-to 10-membered aryl.
1317. The agent of embodiment 1315, wherein R is optionally substituted phenyl.
1318. The agent of embodiment 1315, wherein R is phenyl.
1319. The agent of embodiment 1315, wherein R is optionally substituted naphthyl.
1320. The agent of embodiment 1315, wherein R is naphthyl.
1321. The agent of embodiment 1315, wherein R is an optionally substituted 5-membered heteroaryl having 1 to 4 heteroatoms.
1322. The agent of embodiment 1315, wherein R is an optionally substituted 6 membered heteroaryl having 1 to 4 heteroatoms.
1323. The agent of embodiment 1315, wherein R is an optionally substituted 9-membered bicyclic heteroaryl having 1 to 4 heteroatoms.
1324. The agent of embodiment 1315, wherein R is an optionally substituted 10 membered bicyclic heteroaryl having 1 to 4 heteroatoms.
1325. The agent of any one of embodiments 1321-1324, wherein the heteroatom is nitrogen.
1326. The agent of any one of embodiments 1321-1324, wherein the heteroatom is oxygen.
1327. The agent of any one of embodiments 1321-1324, wherein the heteroatom is sulfur.
1328. The agent of any one of embodiments 1321-1324, wherein the heteroaryl has only one heteroatom.
1329. The agent of embodiment 1328, wherein the heteroatom is nitrogen.
1330. The agent of embodiment 1328, wherein the heteroatom is oxygen.
1331. The agent of embodiment 1328, wherein the heteroatom is sulfur.
1332. The agent of any one of embodiments 1315-1331, wherein L a Is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1333. The agent of any one of embodiments 1315-1331, wherein L a Is an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1334. The agent of any one of embodiments 1315-1331, wherein L a Is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1335. The agent of embodiment 1332, wherein L a Is optionally substituted- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1336. The agent of embodiment 1332, wherein L a Is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1337. The agent of embodiment 1332, wherein L a is-CH 2 -。
1338. The agent of any one of the preceding embodiments, wherein X 13 Comprises an optionally substituted 8-to 10-membered bicyclic aromatic group.
1339. The agent of any one of the preceding embodiments, wherein X 13 Comprises an optionally substituted 9 membered bicyclic heteroaryl group having 1 to 3 heteroatoms.
1340. The agent of any one of the preceding embodiments, wherein X 13 Is BtzA.
1341. The agent of any one of embodiments 1 to 1338, wherein X 13 Is 2NapA.
1342. The agent of any one of embodiments 1309, wherein the aromatic group is phenyl.
1343. The agent of any of embodiments 1342, wherein X 13 34ClF.
1344. The agent of any one of embodiments 1 to 1308, wherein X 13 Selected from BztA,34ClF,2NapA,3BrF, and 34MeF.
1345. The agent of any one of embodiments 1 to 1308, wherein X 13 Is 3Thi.
1346. The agent of any one of embodiments 1 to 1308, wherein X 13 Is Phe.
1347. The agent of any one of embodiments 1 to 1308, wherein X 13 Is GlnR.
1348. The agent of any one of embodiments 1 to 1308, wherein X 13 34MeF.
1349. The agent of any one of embodiments 1 to 1308, wherein X 13 Is 2NapA.
1350. The agent of any one of embodiments 1 to 1308, wherein X 13 Is Lys.
1351. The agent of any one of the preceding embodiments, wherein X 13 Interact with Gln379 of beta-catenin or with its corresponding amino acid residue.
1352. The agent of any one of the preceding embodiments, wherein X 13 Interactions with Leu382 of β -catenin or with its corresponding amino acid residue.
1353. The agent of any one of the preceding embodiments, wherein X 13 Interact with Val416 of β -catenin or with its corresponding amino acid residue.
1354. The agent of any one of the preceding embodiments, wherein X 13 Interact with Asn415 of β -catenin or with its corresponding amino acid residue.
1355. The agent of any one of the preceding embodiments, wherein X 13 Interact with Trp383 of beta-catenin or with its corresponding amino acid residue.
1356. The agent of any one of the preceding embodiments, wherein X 14 Are not stapled.
1357. The agent of any one of the preceding embodiments, wherein X 14 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)-。
1358. The agent of embodiment 1357, wherein R a1 is-H.
1359. The agent of any one of embodiments 1357 to 1358, wherein R a3 is-H.
1360. The agent of any one of embodiments 1357 to 1358, wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
1361. The agent of any one of embodiments 1357 to 1360, wherein L a1 Is a covalent bond.
1362. The agent of any one of embodiments 1357 to 1361, wherein L a2 Is a covalent bond.
1363. The agent of any one of embodiments 1357 through 1362, wherein R a2 is-L' -R.
1364. The agent of any one of embodiments 1357 through 1362, wherein R a2 is-L' -Cy-R.
1365. The agent of any one of embodiments 1357 through 1362, wherein R a2 is-L' -C (O) OR.
1366. The agent of any one of embodiments 1357 through 1362, wherein R a2 is-L '-C (O) N (R') 2 。
1367. The agent of any one of embodiments 1357 through 1362, wherein R a2 is-L "-C (O) N (R) 2 。
1368. The agent of any one of embodiments 1359-1367, wherein R is hydrogen or optionally substituted C 1-10 Aliphatic series.
1369. The agent of any one of embodiments 1359-1367, wherein R is hydrogen.
1370. The agent of any one of embodiments 1359-1367, wherein R is optionally substituted C 1-10 Aliphatic series.
1371. The agent of any one of embodiments 1359-1367, wherein R is C 1-10 Aliphatic series.
1372. The agent of any one of embodiments 1359-1367, wherein R is C 1-10 An alkyl group.
1373. The agent of any one of embodiments 1357 through 1362, wherein R a2 is-L' -OH.
1374. The agent of any one of embodiments 1357 to 1373, wherein L "is a covalent bond or optionally is takenSubstituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1375. The agent of any one of embodiments 1357 through 1373, wherein L "is an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1376. The agent of any one of embodiments 1357 through 1373, wherein L "is a divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1377. The agent of any one of embodiments 1357 through 1373, wherein L "is optionally substituted- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1378. The agent of any one of embodiments 1357 through 1373, wherein L "is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1379. The agent of any one of embodiments 1 to 1355, wherein X 14 Is GlnR.
1380. The agent of any one of embodiments 1 to 1355, wherein X 14 Is BztA.
1381. The agent of any one of embodiments 1 to 1355, wherein X 14 Is sA.
1382. The agent of any one of embodiments 1 to 1355, wherein X 14 34ClF.
1383. The agent of any one of embodiments 1 to 1355, wherein X 14 Is Cys.
1384. The agent of any one of embodiments 1 to 1355, wherein X 14 Is Ala.
1385. The agent of any one of embodiments 1 to 1355, wherein X 14 Is Lys.
1386. The agent of any one of embodiments 1 to 1355, whereinX 14 Is AsnR.
1387. The agent of any one of embodiments 1 to 1355, wherein X 14 Is aMeC.
1388. The agent of any one of embodiments 1 to 1355, wherein X 14 Is pyrS2.
1389. The agent of any one of embodiments 1 to 1355, wherein X 14 Is hGLnR.
1390. The agent of any one of embodiments 1 to 1355, wherein X 14 Is 3Thi.
1391. The agent of any one of embodiments 1 to 1355, wherein X 14 Is Lys.
1392. The agent of any one of embodiments 1 to 1355, wherein X 14 Is Gln.
1393. The agent of any one of the preceding embodiments, wherein X 14 Comprising a C-terminal group.
1394. The agent of any one of the preceding embodiments, wherein X 15 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)-。
1395. The medicament of embodiment 1394, wherein R a1 is-H.
1396. The agent of any one of embodiments 1394 to 1395, wherein R a3 is-H.
1397. The agent of any one of embodiments 1394 to 1395, wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
1398. The agent of any one of embodiments 1394 to 1397, wherein L a1 Is a covalent bond.
1399. The medicament of any one of embodiments 1394 to 1398, wherein L a2 Is a covalent bond.
1400. The agent of any one of embodiments 1394 to 1399, wherein R a2 is-L' -R.
1401. The agent of any one of embodiments 1394 to 1399, wherein R a2 is-L' -Cy-R.
1402. Embodiments 1394 to 1399, wherein R a2 is-L' -C (O) OR.
1403. The agent of any one of embodiments 1394 to 1399, wherein R a2 is-L '-C (O) N (R') 2 。
1404. The agent of any one of embodiments 1394 to 1399, wherein R a2 is-L "-C (O) N (R) 2 。
1405. The agent of any of embodiments 1400-1404 wherein R is hydrogen or optionally substituted C 1-10 Aliphatic series.
1406. The agent of any one of embodiments 1400-1404, wherein R is hydrogen.
1407. The agent of any of embodiments 1400-1404, wherein R is optionally substituted C 1-10 Aliphatic series.
1408. The agent of any of embodiments 1400-1404, wherein R is C 1-10 Aliphatic series.
1409. The agent of any of embodiments 1400-1404, wherein R is C 1-10 An alkyl group.
1410. The agent of any one of embodiments 1394 to 1399, wherein R a2 is-L' -OH.
1411. The agent of any one of embodiments 1394-1410, wherein L "is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1412. The agent of any one of embodiments 1394-1410, wherein L "is an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1413. The agent of any of embodiments 1394-1410, wherein L "is a divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R '), -C (O) -, -C (O) N (R') -, or-N (R) ') C (O) O-substitution.
1414. The agent of any of embodiments 1394-1410, wherein L "is a divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1415. The agent of any of embodiments 1394-1410, wherein L "is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1416. The agent of any one of the preceding embodiments, wherein p15 is 1.
1417. The agent of any one of the preceding embodiments, wherein X 15 Selected from Ala, lcu, val, aib, morphNva, thr, dA, dLeu, [ biotin PEG8]]Lys, glu, and AzLys.
1418. The agent of any one of the preceding embodiments, wherein X 15 Comprising hydrophobic side chains.
1419. The agent of any one of the preceding embodiments, wherein X 15 Is C as a side chain 1-10 An alkyl group.
1420. The agent of any one of the preceding embodiments, wherein X 15 Is Ala.
1421. The agent of any one of the preceding embodiments, wherein X 15 Is optionally substituted or labelled Lys.
1422. The agent of any one of embodiments 1 to 1393, wherein X 15 Is that
Ala, glnR, leu, val, ser, thr,3Thi, bztA, aib, morphNva, dA, dLeu, pro, phe, [ biotin PEG8] Lys, throl, glu, azLys, npg, trp, iyr, lys, prool, alaol, gly, dPro, asn, gln, ala_D3, [ mPEG4] Lys, [ mPEG8] Lys, or [ mPEG16] Lys.
1423. The agent of any one of embodiments 1 to 1393, wherein X 15 Is Ala.
1424. The agent of any one of embodiments 1 to 1393, wherein X 15 Is optionally substituted or labelled Lys.
1425. Implementation of the embodimentsThe agent of any one of schemes 1-1393, wherein X 15 Is GlnR.
1426. The agent of any one of embodiments 1 to 1393, wherein X 15 Is Leu.
1427. The agent of any one of embodiments 1 to 1393, wherein X 15 Is Val.
1428. The agent of any one of embodiments 1 to 1393, wherein X 15 Is Ser.
1429. The agent of any one of embodiments 1 to 1393, wherein X 15 Is Thr.
1430. The agent of any one of embodiments 1 to 1393, wherein X 15 Is 3Thi.
1431. The agent of any one of embodiments 1 to 1393, wherein X 15 Is BztA.
1432. The agent of any one of the preceding embodiments, wherein X 15 Comprising a C-terminal group.
1433. The agent of any one of embodiments 1-1393, wherein p15 is 0.
1434. The agent of any one of the preceding embodiments, wherein p16 is 1.
1435. The agent of any one of the preceding embodiments, wherein X 16 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)-。
1436. The agent of embodiment 1435 wherein R a1 is-H.
1437. The agent of any one of embodiments 1435 through 1436 wherein R a3 is-H.
1438. The agent of any one of embodiments 1435 through 1436 wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
1439. The agent of any one of embodiments 1435-1438 wherein L a1 Is a covalent bond.
1440. The agent of any one of embodiments 1435 to 1439 wherein L a2 Is a covalent bond.
1441. Embodiments 1435 through 1440, wherein R a2 is-L' -R.
1442. The agent of any one of embodiments 1435 through 1440 wherein R a2 is-L' -Cy-R.
1443. The agent of any one of embodiments 1435 through 1440 wherein R a2 is-L' -C (O) OR.
1444. The agent of any one of embodiments 1435 through 1440 wherein R a2 is-L '-C (O) N (R') 2 。
1445. The agent of any one of embodiments 1435 through 1440 wherein R a2 is-L "-C (O) N (R) 2 。
1446. The agent of any one of embodiments 1441-1445, wherein R is hydrogen or optionally substituted C 1-10 Aliphatic series.
1447. The agent of any one of embodiments 1441-1445, wherein R is hydrogen.
1448. The agent of any one of embodiments 1441-1445, wherein R is optionally substituted C 1-10 Aliphatic series.
1449. The agent of any one of embodiments 1441-1445, wherein R is C 1-10 Aliphatic series.
1450. The agent of any one of embodiments 1441-1445, wherein R is C 1-10 An alkyl group.
1451. The agent of any one of embodiments 1435 through 1440 wherein R a2 is-L' -OH.
1452. The agent of any one of embodiments 1435-1451 wherein L "is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1453. The agent of any one of embodiments 1435 through 1451 wherein L "is an optionally substituted divalent C 1-10 Aliphatic wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-And (3) replacing.
1454. The agent of any one of embodiments 1435 through 1451 wherein L "is a divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1455. The agent of any one of embodiments 1435 through 1451 wherein L "is a divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1456. The agent of any one of embodiments 1435-1451 wherein L "is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1457. The agent of any one of embodiments 1 to 1434 wherein X 16 Selected from Ser, ala, glu, aib, asp, thr, and aThr.
1458. The agent of any one of embodiments 1 to 1434 wherein X 16 Is Ala, ser, glu, glnR, bztA, thr, aib, asp, lys, aThr, val, or Arg.
1459. The agent of any one of the preceding embodiments, wherein X 16 Comprising a C-terminal group.
1460. The agent of any one of embodiments 1-1433 wherein p16 is 0.
1461. The agent of any one of the preceding embodiments, wherein p17 is 1.
1462. The agent of any one of the preceding embodiments, wherein X 17 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)-。
1463. The agent of embodiment 1462, wherein R a1 is-H.
1464. The agent of any one of embodiments 1462-1463, wherein R a3 is-H.
1465. The agent of any one of embodiments 1462-1463, wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
1466. The agent of any one of embodiments 1462-1465, wherein L a1 Is a covalent bond.
1467. The agent of any one of embodiments 1462-1466, wherein L a2 Is a covalent bond.
1468. The agent of any one of embodiments 1462-1467, wherein R a2 is-L' -R.
1469. The agent of any one of embodiments 1462-1467, wherein R a2 is-L' -Cv-R.
1470. The agent of any one of embodiments 1462-1467, wherein R a2 is-L' -C (O) OR.
1471. The agent of any one of embodiments 1462-1467, wherein R a2 is-L '-C (O) N (R') 2 。
1472. The agent of any one of embodiments 1462-1467, wherein R a2 is-L "-C (O) N (R) 2 。
1473. The agent of any one of embodiments 1468-1472, wherein R is hydrogen or optionally substituted C 1-10 Aliphatic series.
1474. The agent of any one of embodiments 1468-1472, wherein R is hydrogen.
1475. The agent of any one of embodiments 1468-1472, wherein R is optionally substituted C 1-10 Aliphatic series.
1476. The agent of any one of embodiments 1468-1472, wherein R is C 1-10 Aliphatic series.
1477. The agent of any one of embodiments 1468-1472, wherein R is C 1-10 An alkyl group.
1478. The agent of any one of embodiments 1462-1467, wherein R a2 is-L' -OH.
1479. The agent of any one of embodiments 1462-1478, wherein L "is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1480. The agent of any one of embodiments 1462-1478, wherein L "is an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1481. The agent of any one of embodiments 1462-1478, wherein L "is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1482. The agent of any one of embodiments 1462-1478, wherein L "is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1483. The agent of any one of embodiments 1462-1478, wherein L "is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1484. The agent of any one of embodiments 1 to 1460, wherein X 17 Is Ala, leu, glnR, glnR, pro, thr, val, lys, arg, [ Ac ]]Lys，[mPEG4]Lys，[mPEG8]Lys, or [ mPWG16]Lys。
1485. The agent of any one of embodiments 1 to 1460, wherein X 17 Selected from Ala and Leu.
1486. The agent of any one of the preceding embodiments, wherein X 17 Comprising a C-terminal group.
1487. The agent of any one of embodiments 1-1460, wherein p17 is 0.
1488. The agent of any one of the preceding embodiments, wherein p18 is 1.
1489. The agent of any one of the preceding embodiments, wherein X 18 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)-。
1490. The agent of embodiment 1489 wherein R a1 is-H.
1491. Description of the embodimentsThe agent of any one of 1489-1490 wherein R a3 is-H.
1492. The agent of any one of embodiments 1489-1490 wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
1493. The agent of any one of embodiments 1489-1492 wherein L a1 Is a covalent bond.
1494. The agent of any one of embodiments 1489-1493 wherein L a2 Is a covalent bond.
1495. The agent of any one of embodiments 1489-1494 wherein R a2 is-L' -R.
1496. The agent of any one of embodiments 1489-1494 wherein R a2 is-L' -Cy-R.
1497. The agent of any one of embodiments 1489-1494 wherein R a2 is-L' -C (O) OR.
1498. The agent of any one of embodiments 1489-1494 wherein R a2 is-L '-C (O) N (R') 2 。
1499. The agent of any one of embodiments 1489-1494 wherein R a2 is-L "-C (O) N (R) 2 。
1500. The agent of any one of embodiments 1495-1499 wherein R is hydrogen or optionally substituted C 1-10 Aliphatic series.
1501. The agent of any one of embodiments 1495 to 1499 wherein R is hydrogen.
1502. The agent of any one of embodiments 1495-1499 wherein R is optionally substituted C 1-10 Aliphatic series.
1503. The agent of any one of embodiments 1495-1499 wherein R is C 1-10 Aliphatic series.
1504. The agent of any one of embodiments 1495-1499 wherein R is C 1-10 An alkyl group.
1505. The agent of any one of embodiments 1489-1494 wherein R a2 is-L' -OH.
1506. Embodiments 1489 to 1505, wherein L "is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1507. The agent of any of embodiments 1489-1505 wherein L "is an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1508. The agent of any of embodiments 1489-1505 wherein L "is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1509. The agent of any of embodiments 1489-1505 wherein L "is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1510. The agent of any of embodiments 1489-1505 wherein L "is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1511. The agent of any one of embodiments 1 through 1487 wherein X 18 Is Ala, pro, leu, [ Ac ]]Lys，[mPEG8]Lys，[mPEG4]Lys，[mPEG16]Lys，Thr，[mPEG37]Lys，[PEG4triPEG16]Lys，[PEG-4triPEG36]Lys, or GinR.
1512. The agent of any one of embodiments 1 through 1487 wherein X 18 Is Ala.
1513. The agent of any one of the preceding embodiments, wherein X 18 Comprising a C-terminal group.
1514. The agent of any one of embodiments 1-1487 wherein p18 is 0.
1515. The agent of any one of the preceding embodiments, wherein p19 is 1.
1516. Any of the foregoing embodimentsThe agent of claim, wherein X 19 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)-。
1517. The agent of embodiment 1516 wherein R a1 is-H.
1518. The agent of any one of embodiments 1516-1517, wherein R a3 is-H.
1519. The agent of any one of embodiments 1516-1517, wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
1520. The agent of any one of embodiments 1516-1519, wherein L a1 Is a covalent bond.
1521. The agent of any one of embodiments 1516-1520, wherein L a2 Is a covalent bond.
1522. The agent of any one of embodiments 1516-1521 wherein R a2 is-L' -R.
1523. The agent of any one of embodiments 1516-1521 wherein R a2 is-L' -Cy-R.
1524. The agent of any one of embodiments 1516-1521 wherein R a2 is-L' -C (O) OR.
1525. The agent of any one of embodiments 1516-1521 wherein R a2 is-L '-C (O) N (R') 2 。
1526. The agent of any one of embodiments 1516-1521 wherein R a2 is-L "-C (O) N (R) 2 。
1527. The agent of any one of embodiments 1522-1526 wherein R is hydrogen or optionally substituted C 1-10 Aliphatic series.
1528. The agent of any one of embodiments 1522-1526 wherein R is hydrogen.
1529. The agent of any one of embodiments 1522-1526 wherein R is optionally substituted C 1-10 Aliphatic series.
1530. The agent of any one of embodiments 1522-1526 wherein R is C 1-10 Aliphatic series.
1531. The agent of any one of embodiments 1522-1526 wherein R is C 1-10 An alkyl group.
1532. The agent of any one of embodiments 1516-1521 wherein R a2 is-L' -OH.
1533. The agent of any one of embodiments 1516-1532, wherein L "is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1534. The agent of any one of embodiments 1516-1532, wherein L "is an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1535. The agent of any one of embodiments 1516-1532, wherein L "is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or tri-N (R ') C (O) O-substitution.
1536. The agent of any one of embodiments 1516-1532, wherein L "is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1537. The agent of any one of embodiments 1516-1532, wherein L "is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1538. The agent of any one of embodiments 1 to 1514, wherein X 19 Is Ala, leu, thr, val, or Pro.
1539. The agent of any one of embodiments 1 through 1487 wherein X 19 Is Ala.
1540. The agent of any one of the preceding embodiments, wherein X 19 Comprising a C-terminal group.
1541. The agent of any one of embodiments 1-1487 wherein p19 is 0.
1542. The agent of any one of the preceding embodiments, wherein p20 is 1.
1543. The agent of any one of the preceding embodiments, wherein X 20 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)-。
1544. The agent of embodiment 1543 wherein R a1 is-H.
1545. The agent of any one of embodiments 1543-1544, wherein R a3 is-H.
1546. The agent of any one of embodiments 1543-1544, wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
1547. The agent of any one of embodiments 1543-1546, wherein L a1 Is a covalent bond.
1548. The agent of any one of embodiments 1543-1547, wherein L a2 Is a covalent bond.
1549. The agent of any one of embodiments 1543-1548, wherein R a2 is-L' -R.
1550. The agent of any one of embodiments 1543-1548, wherein R a2 is-L' -Cy-R.
1551. The agent of any one of embodiments 1543-1548, wherein R a2 is-L' -C (O) OR.
1552. The agent of any one of embodiments 1543-1548, wherein R a2 is-L '-C (O) N (R') 2 。
1553. The agent of any one of embodiments 1543-1548, wherein R a2 is-L "-C (O) N (R) 2 。
1554. The agent of any one of embodiments 1549 through 1553, wherein R is hydrogen or optionally substituted C 1-10 Aliphatic series.
1555. The agent of any one of embodiments 1549 to 1553, wherein R is hydrogen.
1556. The agent of any one of embodiments 1549 through 1553, wherein R is optionally substitutedC 1-10 Aliphatic series.
1557. The agent of any one of embodiments 1549 through 1553, wherein R is C 1-10 Aliphatic series.
1558. The agent of any one of embodiments 1549 through 1553, wherein R is C 1-10 An alkyl group.
1559. The agent of any one of embodiments 1543-1548, wherein R a2 is-L' -OH.
1560. The agent of any one of embodiments 1543 through 1559, wherein L "is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1561. The agent of any one of embodiments 1543 through 1559, wherein L "is an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1562. The agent of any of embodiments 1543 through 1559, wherein L "is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1563. The agent of any of embodiments 1543 through 1559, wherein L "is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1564. The agent of any of embodiments 1543 through 1559, wherein L "is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1565. The agent of any one of embodiments 1 to 1541, wherein X 20 Is Ala, leu, lys, nLeu, val, or Arg.
1566. The agent of any one of embodiments 1 to 1541, wherein X 20 Is Ala.
1567. The agent of any one of the preceding embodiments, wherein X 20 Comprising a C-terminal group.
1568. The agent of any one of embodiments 1 to 1541, wherein p20 is 0.
1569. The agent of any one of the preceding embodiments, wherein p21 is 1.
1570. The agent of any one of the preceding embodiments, wherein X 21 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)-。
1571. The agent of embodiment 1570, wherein R a1 is-H.
1572. The agent of any one of embodiments 1570 to 1571, wherein R a3 is-H.
1573. The agent of any one of embodiments 1570 to 1571, wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
1574. The agent of any one of embodiments 1570 to 1573, wherein L a1 Is a covalent bond.
1575. The agent of any one of embodiments 1570 to 1574, wherein L a2 Is a covalent bond.
1576. The agent of any one of embodiments 1570 to 1575, wherein R a2 is-L' -R.
1577. The agent of any one of embodiments 1570 to 1575, wherein R a2 is-L' -Cy-R.
1578. The agent of any one of embodiments 1570 to 1575, wherein R a2 is-L' -C (O) OR.
1579. The agent of any one of embodiments 1570 to 1575, wherein R a2 is-L '-C (O) N (R') 2 。
1580. The agent of any one of embodiments 1570 to 1575, wherein R a2 is-L "-C (O) N (R) 2 。
1581. The agent of any one of embodiments 1576-1580, wherein R is hydrogen or optionally substituted C 1-10 Aliphatic series.
1582. The agent of any one of embodiments 1576-1580, wherein R is hydrogen.
1583. The agent of any one of embodiments 1576-1580, wherein R is optionally substituted C 1-10 Aliphatic series.
1584. The agent of any one of embodiments 1576-1580, wherein R is C 1-10 Aliphatic series.
1585. The agent of any one of embodiments 1576-1580, wherein R is C 1-10 An alkyl group.
1586. The agent of any one of embodiments 1570 to 1575, wherein R a2 is-L' -OH.
1587. The agent of any one of embodiments 1570-1586, wherein L "is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1588. The agent of any one of embodiments 1570-1586, wherein L "is an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1589. The agent of any one of embodiments 1570-1586, wherein L "is a divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1590. The agent of any one of embodiments 1570-1586, wherein L "is a divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or tri-N (R ') C (O) O-substitution.
1591. The agent of any one of embodiments 1570-1586, wherein L "is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1592. The agent of any one of embodiments 1 to 1568 wherein X 21 Is Ala, leu, lys, nLeu, val, or Arg.
1593. The agent of any one of embodiments 1 to 1568 wherein X 21 Is Ala.
1594. The agent of any one of the preceding embodiments, wherein X 21 Comprising a C-terminal group.
1595. The agent of any one of embodiments 1-1568, wherein p21 is 0.
1596. The agent of any one of the preceding embodiments, wherein p22 is 1.
1597. The agent of any one of the preceding embodiments, wherein X 22 is-N (R) a1 )-L a1 -C(R a2 )(Ra3)-L a2 -C(O)-。
1598. The agent of embodiment 1597, wherein R a1 is-H.
1599. The agent of any one of embodiments 1597 to 1598, wherein R a3 is-H.
1600. The agent of any one of embodiments 1597 to 1598, wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
1601. The agent of any one of embodiments 1597 to 1600, wherein L a1 Is a covalent bond.
1602. The agent of any one of embodiments 1597 to 1601, wherein L a2 Is a covalent bond.
1603. The agent of any one of embodiments 1597 to 1602, wherein R a2 is-L' -R.
1604. The agent of any one of embodiments 1597 to 1602, wherein R a2 is-L' -Cy-R.
1605. The agent of any one of embodiments 1597 to 1602, wherein R a2 is-L' -C (O) OR.
1606. The agent of any one of embodiments 1597 to 1602, wherein R a2 is-L '-C (O) N (R') 2 。
1607. The agent of any one of embodiments 1597 to 1602, wherein R a2 is-L "-C (O) N (R) 2 。
1608. The agent of any of embodiments 1603 to 1607, wherein R is hydrogen or optionally substituted C 1-10 Aliphatic series.
1609. The agent of any one of embodiments 1603 to 1607, wherein R is hydrogen.
1610. The agent of any of embodiments 1603 to 1607, wherein R is optionally substituted C 1-10 Aliphatic series.
1611. The agent of any of embodiments 1603 to 1607, wherein R is C 1-10 Aliphatic series.
1612. The agent of any of embodiments 1603 to 1607, wherein R is C 1-10 An alkyl group.
1613. The agent of any one of embodiments 1597 to 1602, wherein R a2 is-L' -OH.
1614. The agent of any one of embodiments 1597 to 1613, wherein L "is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1615. The agent of any one of embodiments 1597 to 1613, wherein L "is an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1616. The agent of any one of embodiments 1597 to 1613, wherein L "is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1617. The agent of any one of embodiments 1597 to 1613, wherein L "is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1618. Any of embodiments 1597 to 1613The agent, wherein L' is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1619. The agent of any one of embodiments 1 to 1595, wherein X 22 Is Lys.
1620. The agent of any one of the preceding embodiments, wherein X 22 Comprising a C-terminal group.
1621. The agent of any one of embodiments 1 to 1595, wherein p22 is 0.
1622. The agent of any one of the preceding embodiments, wherein p23 is 1.
1623. The agent of any one of the preceding embodiments, wherein X 23 is-N (R) a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)-。
1624. The agent of embodiment 1623, wherein R a1 is-H.
1625. The agent of any one of embodiments 1623-1624, wherein R a3 is-H.
1626. The agent of any one of embodiments 1623-1624, wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
1627. The agent of any one of embodiments 1623-1626, wherein L a1 Is a covalent bond.
1628. The agent of any one of embodiments 1623-1627, wherein L a2 Is a covalent bond.
1629. The agent of any one of embodiments 1623-1628, wherein R a2 is-L' -R.
1630. The agent of any one of embodiments 1623-1628, wherein R a2 is-L' -Cy-R.
1631. The agent of any one of embodiments 1623-1628, wherein R a2 is-L' -C (O) OR.
1632. The agent of any one of embodiments 1623-1628, wherein R a2 is-L '-C (O) N (R') 2 。
1633. The agent of any one of embodiments 1623-1628, wherein R a2 is-L "-C (O) N (R) 2 。
1634. The agent of any one of embodiments 1629-1633, wherein R is hydrogen or optionally substituted C 1-10 Aliphatic series.
1635. The agent of any one of embodiments 1629-1633, wherein R is hydrogen.
1636. The agent of any one of embodiments 1629-1633, wherein R is optionally substituted C 1-10 Aliphatic series.
1637. The agent of any one of embodiments 1629-1633, wherein R is C 1-10 Aliphatic series.
1638. The agent of any one of embodiments 1629-1633, wherein R is C 1-10 An alkyl group.
1639. The agent of any one of embodiments 1623-1628, wherein R a2 is-L' -OH.
1640. The agent of any one of embodiments 1623-1639, wherein L "is a covalent bond or an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1641. The agent of any one of embodiments 1623-1639, wherein L "is an optionally substituted divalent C 1-10 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -Cy-, -N (R '), -C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1642. The agent of any one of embodiments 1623-1639, wherein L "is a divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1643. The agent of any one of embodiments 1623-1639, wherein L "is a divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1644. The agent of any one of embodiments 1623-1639, wherein L "is- (CH) 2 ) n-, wherein n is 1, 2, 3, 4, 5 or 6.
1645. The agent of any one of the preceding embodiments, wherein X 23 Comprising a C-terminal group.
1646. The agent of any one of embodiments 1 to 1621, wherein p23 is 0.
1647. The agent of any of the preceding embodiments, wherein the C-terminal group is R C 。
1648. The agent of any one of embodiments 1 to 1646, wherein the C-terminal group is-OH.
1649. The agent of any one of embodiments 1 to 1646, wherein the C-terminal group is-N (R) 2 。
1650. The agent of any one of embodiments 1 to 1646, wherein the C-terminal group is-N (R) 2 Wherein each R is independently-H or optionally substituted C 1-6 Aliphatic series.
1651. The agent of any one of embodiments 1 to 1646, wherein the C-terminal group is-NH 2 。
1652. The agent of any one of embodiments 1 to 1646, wherein the C-terminal group is-NHMe.
1653. The agent of any one of embodiments 1 to 1646, wherein the C-terminal group is-NHEt.
1654. The agent of any one of embodiments 1 to 1646, wherein the C-terminal group is Serol.
1655. The agent of any one of embodiments 1 to 1646, wherein the C-terminal group is dalacol.
1656. The agent of any one of the preceding embodiments, wherein the peptide comprises a hydrocarbon staple-like structure.
1657. The agent of any one of the preceding embodiments, wherein the peptide comprises a non-hydrocarbon staple-like structure.
1658. The agent of any one of the preceding embodiments, wherein the peptide comprises a staple-like structure, the chain of the staple-like structure comprising-N (R ') -or-O-C (O) -N (R') -.
1659. The agent of any one of the preceding embodiments, wherein the peptide has the following structure or salt thereof:
R N -[X] p -[X 0 ] p0 X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 -[X] p’ -R C ，
wherein:
each X is independently an amino acid residue;
each p and p' is independently 0 to 10;
R N independently a peptide, amino protecting group or R' -L RN -；
R C Independently a peptide, a carboxyl protecting group, -L RC -R’、-O-L RC -R 'or-N (R') -L RC -R’；
L RN And L RC Each independently is L;
each L is independently a covalent bond, or an optionally substituted divalent C 1 -C 25 Aliphatic or heteroaliphatic groups having from 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -，-Cy-,-O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-,-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-;
each-Cy-is independently an optionally substituted divalent 3-to 30-membered monocyclic, bicyclic, or polycyclic ring having 0 to 10 heteroatoms;
each R' is independently-R, -C (O) R, -CO 2 R or-SO 2 R；
Each R is independently-H or an optionally substituted group selected from: c (C) 1-30 Aliphatic, havingC of 1 to 10 hetero atoms 1-30 Heteroaliphatic, C 6-30 Aryl, C 6-30 Arylaliphatic, C having 1 to 10 heteroatoms 6-30 Aryl heteroaliphatics, 5 to 30 membered heteroaryl groups having 1 to 10 heteroatoms, and 3 to 30 membered heterocyclyl groups having 1 to 10 heteroatoms, or
The two R groups optionally and independently together form a covalent bond, or:
two or more R groups on the same atom optionally and independently taken together with the atom form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the atom; or alternatively
Two or more R groups on two or more atoms optionally and independently together with their intervening atoms form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the intervening atoms.
1660. The agent of any one of the preceding embodiments, wherein p is 0.
1661. The agent of any one of the preceding embodiments, wherein p is 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10).
1662. The agent of any one of embodiments 1 to 1659, wherein p is 1.
1663. The agent of any one of embodiments 1 to 1659, wherein p is 2.
1664. The agent of any one of embodiments 1 to 1659, wherein p is 3.
1665. The agent of any one of embodiments 1 to 1659, wherein p is 4.
1666. The agent of any one of embodiments 1 to 1659, wherein p is 5.
1667. The agent of any one of the preceding embodiments, wherein p' is 0.
1668. The agent of any one of embodiments 1-1666, wherein p' is 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10).
1669. The agent of any one of embodiments 1 to 1666 wherein p' is 2.
1670. The agent of any one of embodiments 1 to 1666 wherein p' is 3.
1671. The agent of any one of embodiments 1 to 1666 wherein p' is 4.
1672. The agent of any one of embodiments 1 to 1666 wherein p' is 5.
1673. The agent of any one of the preceding embodiments, wherein R N is-C (O) R.
1674. The agent of any one of the preceding embodiments, wherein R N Is Ac.
1675. The agent of any one of embodiments 1 to 1672, wherein R N Is that
Ac, NPyroR3,5 hexenyl, 4 pentenyl, bua, C3a, cpc, cbc, cypCO, bnc, CF3CO,2PyCypCO,4THPCO, isobutyryl, ts,15pyraPy,2PyBu,4PymCO,4PyPrpc,3lAPAc,4MePipzPrpC, mePipAc, meImid4SO2, bzAm2O allyl, hex,2PyzCO,3Phc3, meOPr, lithocholic acid, 2FPhc, phC, meSO2, isovaleryl, etHNCO, tzPyr,8IAP,3PydCO,2PymCO,5PymCO,1Imidac,2F2PyAc,21APAc,124TriPr,6QuiAc,3PyAc,123TriAc,1 pyrazole Ac,3PyPrpc,5PymAc,1 pyrone Ac,124TriAc, me2NAc,8QuiSO2, mPEG4, mPEG8, mPEG16 or mPEG24.
1676. The agent of any one of embodiments 1 to 1672, wherein R N Is 4 pentenyl.
1677. The agent of any one of embodiments 1 to 1672, wherein R N Is 5 hexenyl.
1678. The agent of any one of embodiments 1 to 1672, wherein R N Is BzAm2O allyl.
1679. The agent of any one of the preceding embodiments, wherein R C is-N (R') 2 。
1680. The agent of any one of the preceding embodiments, wherein R C is-N (R) 2 。
1681. Any of embodiments 1 through 1680The agent of claim, wherein R C is-NH 2 。
1682. The agent of any one of embodiments 1 to 1680, wherein R C is-NHEt.
1683. The agent of any one of embodiments 1 to 1680, wherein R C is-Alaol, wherein the amino group of-Alaol is bonded with the last-C (O) -bond of the peptide skeleton (R C Is that)。
1684. The agent of any one of embodiments 1 to 1680, wherein R C is-dAlol, wherein the amino group of-dAlol is bonded to the last-C (O) -bond of the peptide backbone (R C Is that)。
1685. The agent of any one of embodiments 1 to 1680, wherein R C is-Prool, wherein the amino group of-Prool is bonded to the last-C (O) -bond of the peptide backbone (R C Is that)。
1686. The agent of any one of embodiments 1 to 1680, wherein R C is-Throol, wherein the amino group of-Throol is bonded to the last-C (O) -bond of the peptide backbone (R C Is that)。
1687. The agent of any one of embodiments 1 to 1680, wherein R C is-Serol, wherein the amino group of-Serol is bonded to the last-C (O) -of the peptide backbone (R C Is that)。
1688. The agent of any one of embodiments 1 to 1678, wherein R C is-OH.
1689. In the foregoing embodimentThe agent of any one of claims, wherein each amino acid residue is independently-N (R a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)-。
1690. The agent of embodiment 1689, wherein R a1 is-H.
1691. The agent of any one of embodiments 1 to 1689, wherein R a1 And R is R a2 Or R is a3 And intervening atoms together form an optionally substituted 3 to 10 (e.g., 3, 4, 5, 6, 7, 8, 9, or 10) membered ring having 0 to 5 heteroatoms in addition to the intervening atoms.
1692. The agent of any one of embodiments 1 to 1689, wherein R a1 And R is R a2 Or R is a3 And intervening atoms together form an optionally substituted 5-to 7-membered ring having no heteroatoms other than the intervening atoms.
1693. The agent of any one of the preceding embodiments, wherein L a1 Is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -Cy-, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1694. The agent of any one of embodiments 1 to 1692, wherein L a1 Is a covalent bond.
1695. The agent of any one of the preceding embodiments, wherein R a2 is-L a -R', wherein L a Is a covalent bond or a divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -Cy-, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1696. The agent of any one of the preceding embodiments, wherein R a3 is-L a -R', wherein L a Is a covalent bond or a divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -Cy-, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1697. The agent of any one of embodiments 1 to 1695, wherein R a3 Is-H。
1698. The agent of any one of embodiments 1 to 1695, wherein R a3 Is optionally substituted C 1-6 Aliphatic series.
1699. The agent of any one of the preceding embodiments, wherein L a2 Is divalent C 1-6 Aliphatic, wherein one or more methylene units are optionally and independently replaced by-O-, -S-, -N (R ') -, C (O) -, -Cy-, -C (O) N (R ') -, or-N (R ') C (O) O-substitution.
1700. The agent of any one of embodiments 1 to 1698, wherein L a2 Is a covalent bond.
1701. The agent of any one of the preceding embodiments, wherein the agent is or comprises a stapled peptide containing a stapled residue at a position referred to as position P.
1702. The agent of embodiment 1701, wherein the stapled residue at position P is stapled with a residue at position p+7.
1703. The agent of any one of embodiments 1701-1702, wherein the stapling residue at position P is stapled with a residue at position P-4.
1704. The agent of any one of embodiments 1701-1702, wherein the stapling residue at position P is stapled with a residue at position P-3.
1705. The agent of any one of embodiments 1701-1702, wherein the stapling residue at position P is stapled with a residue at position P-2.
1706. The agent of any one of embodiments 1701-1705, wherein the stapled peptide comprises a two residue stapled, staple-like structure at positions p+6 and p+10.
1707. The agent of any one of embodiments 1701-1705, wherein the stapled peptide comprises a two residue stapled, staple-like structure at positions p+3 and p+10.
1708. The agent of any one of embodiments 1701-1707, wherein there are three staple-like structures in the stapled peptide.
1709. The agent of any one of embodiments 1701-1707, wherein the stapled peptide comprises a two residue stapled, staple-like structure at positions P-1 and p+3.
1710. The agent of any one of embodiments 1701-1707 and 1709, wherein there are four staple-like structures in the stapled peptide.
1711. The agent of any one of embodiments 1701-1710, wherein the stapled peptide comprises an acidic amino acid residue at position P-2.
1712. The agent of any one of embodiments 1701-1711, wherein the stapled peptide comprises an acidic amino acid residue at position p+1.
1713. The agent of any one of embodiments 1701-1712, wherein the stapled peptide comprises an acidic amino acid residue at position p+2.
1714. The agent of any one of embodiments 1701-1713, wherein the stapled peptide comprises a hydrophobic amino acid residue at position p+4.
1715. The agent of any one of embodiments 1701-1714, wherein the stapled peptide comprises an aromatic amino acid residue at position p+5.
1716. The agent of any one of embodiments 1701-1715, wherein the stapled peptide comprises an aromatic amino acid residue at position p+8.
1717. The agent of any one of embodiments 1701-1716, wherein the stapled peptide comprises an aromatic amino acid residue at position p+9.
1718. The agent of any one of embodiments 1701-1717, wherein position P is position 3.
1719. The agent of any one of embodiments 1701-1717, wherein position P is position 4.
1720. The agent of any one of embodiments 1701-1717, wherein position P is position 5.
1721. The agent of any one of embodiments 1701-1717, wherein position P is position 6.
1722. The agent of any one of embodiments 1701-1717, wherein position P is position 7.
1723. The agent of any one of the preceding embodiments, wherein the peptide forms a structure comprising a helix.
1724. The agent of any one of the preceding embodiments, wherein the peptide binds to β -catenin.
1725. The agent of any one of the preceding embodiments, wherein the peptide binds to β -catenin with the following EC 50: no greater than about 2000nM, or no greater than about 1500nM, or no greater than about 1000nM, or no greater than about 500nM, or no greater than about 300nM, or no greater than about 200nM, or no greater than about 100nM, or no greater than about 75nM, or no greater than about 50nM, or no greater than about 25nM, or no greater than about 10nM, as measured by fluorescence polarization.
1726. The agent of any one of the preceding embodiments, wherein the peptide can compete for β -catenin binding with TCF7, LEF1, TCF7L2, axin1, axin2, or APC, or fragments thereof.
1727. The agent of any one of the preceding embodiments, wherein the peptide and sequence is SEQ ID NO:2 or a sequence comprising SEQ ID NO:2 or a fragment thereof:
1728. the agent of any one of the preceding embodiments, wherein the peptide binds to β -catenin and interacts with one or more residues that are or correspond to: SEQ ID NO:1, or at least three, or at least four, or at least five, or at least six, or at least seven, or at least eight, or at least nine, or at least ten, or at least eleven, or at least twelve, or at least thirteen, or at least fourteen, or at least fifteen, or at least sixteen, or at least seventeen, or at least eighteen, or at least nineteen, or at least twenty of the following amino acid residues at the positions indicated in 1:
a305, Y306, G307, N308, Q309, K312, R342, K345, V346, V349, Q375, R376, Q379, N380, L382, W383, R386, N387, D413, N415, V416, T418, and C419.
1729. The agent of any one of the preceding embodiments, wherein the peptide binds to β -catenin and interacts with one or more residues that are or correspond to: SEQ ID NO:1, or at least three, or at least four, or at least five, or at least six, or at least seven, or at least eight, or at least nine, or at least ten, or at least eleven, or at least twelve, or at least thirteen, or at least fourteen, or at least fifteen, or at least sixteen, or at least seventeen, or at least eighteen, or at least nineteen, or at least twenty of the following amino acid residues at the positions indicated in 1:
a305 Y306, G307, N308, Q309, K312, R342, K345, V346, V349, Q375, Q379, N380, L382, W383, R386, N387, D413, N415, V416, T418, and C419.
1730. The agent of any one of the preceding embodiments, wherein the peptide binds to β -catenin and interacts with one or more residues that are or correspond to: SEQ ID NO:1, or at least three, or at least four, or at least five, or at least six, or at least seven, or at least eight, or at least nine, or at least ten, or at least eleven, or at least twelve, or at least thirteen, or at least fourteen, or at least fifteen, or at least sixteen, or at least seventeen, or at least eighteen, or at least nineteen, or at least twenty of the following amino acid residues at the positions indicated in 1:
A305, Y306, G307, N308, Q309, K312, K345, V346, V349, Q379, N380, L382, W383, R386, N387, D413, N415, V416, T418, and C419.
1731. The agent of any one of the preceding embodiments, wherein the peptide binds to β -catenin and interacts with one or more residues that are or correspond to: SEQ ID NO:1, or at least three, or at least four, or at least five, or at least six, or at least seven of the following amino acid residues at the positions shown in 1: g307 K312, K345, W383, N387, D413, and N415.
1732. The agent of any one of the preceding embodiments, wherein the agent interacts with Y306 of β -catenin or with its corresponding amino acid residue.
1733. The agent of any one of the preceding embodiments, wherein the agent interacts with G307 of β -catenin or with its corresponding amino acid residue.
1734. The agent of any one of the preceding embodiments, wherein the agent interacts with K312 of β -catenin or with the corresponding amino acid residue thereof.
1735. The agent of any one of the preceding embodiments, wherein the agent interacts with K345 of β -catenin or with the corresponding amino acid residue thereof.
1736. The agent of any one of the preceding embodiments, wherein the agent interacts with Q379 of β -catenin or with its corresponding amino acid residue.
1737. The agent of any one of the preceding embodiments, wherein the agent interacts with L382 of β -catenin or with the corresponding amino acid residue thereof.
1738. The agent of any one of the preceding embodiments, wherein the agent interacts with W383 of β -catenin or with the corresponding amino acid residue thereof.
1739. The agent of any one of the preceding embodiments, wherein the agent interacts with N387 of β -catenin or with the corresponding amino acid residue thereof.
1740. The agent of any one of the preceding embodiments, wherein the agent interacts with D413 of β -catenin or with the corresponding amino acid residue thereof.
1741. The agent of any one of the preceding embodiments, wherein the agent interacts with N415 of β -catenin or with the corresponding amino acid residue thereof.
1742. The agent of any one of the preceding embodiments, wherein the agent interacts with V416 of β -catenin or with the corresponding amino acid residue thereof.
1743. The agent of any one of the preceding embodiments, wherein the agent binds to β -catenin at a site other than an axin binding site.
1744. The agent of any one of the preceding embodiments, wherein the agent binds to β -catenin at a site other than a Bcl9 binding site.
1745. The agent of any one of the preceding embodiments, wherein the agent binds to β -catenin at a site other than a TCF binding site.
1746. The agent of any one of the preceding embodiments, wherein the agent is the peptide.
1747. An agent having a structure selected from table E2 or a salt thereof.
1748. An agent having a structure selected from E3 or a salt thereof.
1749. A medicament havingA structure or a salt thereof.
1750. A medicament havingA structure or a salt thereof.
1751. A medicament havingA structure or a salt thereof.
1752. A medicament havingA structure or a salt thereof. />
1753. A medicament havingA structure or a salt thereof.
1754. A medicament havingA structure or a salt thereof.
1755. A medicament havingA structure or a salt thereof. />
1756. A medicament havingA structure or a salt thereof.
1757. A medicament havingA structure or a salt thereof.
1758. A medicament havingA structure or a salt thereof. />
1759. A medicament havingA structure or a salt thereof.
1760. A medicament havingA structure or a salt thereof.
1761. A medicament havingA structure or a salt thereof. />
1762. A medicament having A structure or a salt thereof.
1763. A medicament havingA structure or a salt thereof.
1764. The agent of any one of the preceding embodiments, wherein the double bond of the staple-like structure bonded to the first stapled amino acid counted from the N-terminus is E, the first stapled amino acid being bonded to the staple-like structure having a double bond.
1765. The agent of any one of the preceding embodiments, wherein the double bond of the staple-like structure bonded to the first stapled amino acid counted from the N-terminus is Z, the first stapled amino acid being bonded to the staple-like structure having a double bond.
1766. The agent of any one of the preceding embodiments, wherein the double bond of the staple-like structure bonded to the first stapled amino acid counted from the C-terminus is E, the first stapled amino acid being bonded to the staple-like structure having a double bond.
1767. The agent of any one of the preceding embodiments, wherein the double bond of the staple-like structure bonded to the first stapled amino acid counted from the C-terminus is Z, the first stapled amino acid being bonded to the staple-like structure having a double bond.
1768. The agent of any of the preceding embodiments, wherein the double bond of the (i, i+7) staple-like structure is E.
1769. The agent of any of the preceding embodiments, wherein the double bond of the (i, i+7) staple-like structure is Z.
1770. The agent of any one of the preceding embodiments, wherein the double bond of the (i, i+2), (i, i+3) or (i, i+4) staple-like structure is E.
1771. The agent of any one of the preceding embodiments, wherein the double bond of the (i, i+2), (i, i+3), or (i, i+4) staple structure is Z.
1772. The agent of any one of embodiments 1747-1763, wherein the double bond of the staple-like structure bonded to the first amino acid from the N-terminus is Z.
1773. The agent of any one of embodiments 1747-1771, wherein the double bond of the staple-like structure bonded to amino acid 11 from the N-terminus is E.
1774. The agent of any one of embodiments 1747-1771, wherein the double bond of the staple-like structure bonded to amino acid 11 from the N-terminus is Z.
1775. The agent of any of the preceding embodiments, wherein the carbon atom bonded to both staple-like structures (e.g., in B5) is in the R configuration.
1776. The agent of any of the preceding embodiments, wherein the carbon atom bonded to both staple-like structures (e.g., in B5) is in the S configuration.
1777. An agent having the structure SP-1-1 or a salt thereof.
1778. An agent having the structure SP-1-2 or a salt thereof.
1779. An agent having the structure SP-1-3 or a salt thereof.
1780. An agent having the structure SP-1-4 or a salt thereof.
1781. An agent having the structure SP-1-5 or a salt thereof.
1782. An agent having the structure SP-1-6 or a salt thereof.
1783. An agent having the structure SP-1-7 or a salt thereof.
1784. An agent having the structure SP-1-8 or a salt thereof.
1785. An agent having the structure SP-2-1 or a salt thereof.
1786. An agent having the structure SP-2-2 or a salt thereof.
1787. An agent having the structure SP-2-3 or a salt thereof.
1788. An agent having the structure SP-2-4 or a salt thereof.
1789. An agent having the structure SP-2-5 or a salt thereof.
1790. An agent having the structure SP-2-6 or a salt thereof.
1791. An agent having the structure SP-2-7 or a salt thereof.
1792. An agent having the structure SP-2-8 or a salt thereof.
1793. An agent having the structure SP-3-1 or a salt thereof.
1794. An agent having the structure SP-3-2 or a salt thereof.
1795. An agent having the structure SP-4-1 or a salt thereof.
1796. An agent having the structure SP-4-2 or a salt thereof.
1797. An agent having the structure SP-4-3 or a salt thereof.
1798. An agent having the structure SP-4-4 or a salt thereof.
1799. An agent having the structure SP-4-5 or a salt thereof.
1800. An agent having the structure SP-4-6 or a salt thereof.
1801. An agent having the structure SP-4-7 or a salt thereof.
1802. An agent having the structure SP-4-8 or a salt thereof.
1803. An agent having the structure SP-5-1 or a salt thereof.
1804. An agent having the structure SP-5-2 or a salt thereof.
1805. An agent having the structure SP-5-3 or a salt thereof.
1806. An agent having the structure SP-5-4 or a salt thereof.
1807. An agent having the structure SP-5-5 or a salt thereof.
1808. An agent having the structure SP-5-6 or a salt thereof.
1809. An agent having the structure SP-5-7 or a salt thereof.
1810. An agent having the structure SP-5-8 or a salt thereof.
1811. An agent having an SP-6 structure or a salt thereof.
1812. An agent having the structure SP-7-1 or a salt thereof.
1813. An agent having the structure SP-7-2 or a salt thereof.
1814. An agent having the structure SP-7-3 or a salt thereof.
1815. An agent having the structure SP-7-4 or a salt thereof.
1816. An agent having the structure SP-7-5 or a salt thereof.
1817. An agent having the structure SP-7-6 or a salt thereof.
1818. An agent having the structure SP-7-7 or a salt thereof.
1819. An agent having the structure SP-7-8 or a salt thereof.
1820. An agent having the structure SP-8-1 or a salt thereof.
1821. An agent having the structure SP-8-2 or a salt thereof.
1822. An agent having the structure SP-8-3 or a salt thereof.
1823. An agent having the structure SP-8-4 or a salt thereof.
1824. An agent having the structure SP-8-5 or a salt thereof.
1825. An agent having the structure SP-8-6 or a salt thereof.
1826. An agent having the structure SP-8-7 or a salt thereof.
1827. An agent having an SP-8-8 structure or a salt thereof.
1828. An agent having the structure SP-9-1 or a salt thereof.
1829. An agent having the structure SP-9-2 or a salt thereof.
1830. An agent having the structure SP-9-3 or a salt thereof.
1831. An agent having the structure SP-9-4 or a salt thereof.
1832. An agent having the structure SP-9-5 or a salt thereof.
1833. An agent having the structure SP-9-6 or a salt thereof.
1834. An agent having the structure SP-9-7 or a salt thereof.
1835. An agent having the structure SP-9-8 or a salt thereof.
1836. An agent having the structure SP-10-1 or a salt thereof.
1837. An agent having the structure SP-10-2 or a salt thereof.
1838. An agent having the structure SP-10-3 or a salt thereof.
1839. An agent having the structure SP-10-4 or a salt thereof.
1840. An agent having the structure SP-10-5 or a salt thereof.
1841. An agent having the structure SP-10-6 or a salt thereof.
1842. An agent having the structure SP-10-7 or a salt thereof.
1843. An agent having the structure SP-10-8 or a salt thereof.
1844. An agent having the structure SP-11-1 or a salt thereof.
1845. An agent having the structure SP-11-2 or a salt thereof.
1846. An agent having the structure SP-11-3 or a salt thereof.
1847. An agent having the structure SP-11-4 or a salt thereof.
1848. An agent having the structure SP-11-5 or a salt thereof.
1849. An agent having the structure SP-11-6 or a salt thereof.
1850. An agent having the structure SP-11-7 or a salt thereof.
1851. An agent having the structure SP-11-8 or a salt thereof.
1852. An agent having the structure SP-12-1 or a salt thereof.
1853. An agent having the structure SP-12-2 or a salt thereof.
1854. An agent having the structure SP-12-3 or a salt thereof.
1855. An agent having the structure SP-12-4 or a salt thereof.
1856. An agent having the structure SP-12-5 or a salt thereof.
1857. An agent having the structure SP-12-6 or a salt thereof.
1858. An agent having the structure SP-12-7 or a salt thereof.
1859. An agent having the structure SP-12-8 or a salt thereof.
1860. An agent having the structure SP-13-1 or a salt thereof.
1861. An agent having the structure SP-13-2 or a salt thereof.
1862. An agent having the structure SP-13-3 or a salt thereof.
1863. An agent having the structure SP-13-4 or a salt thereof.
1864. An agent having the structure SP-13-5 or a salt thereof.
1865. An agent having the structure SP-13-6 or a salt thereof.
1866. An agent having the structure SP-13-7 or a salt thereof.
1867. An agent having the structure SP-13-8 or a salt thereof.
1868. An agent having the structure SP-14-1 or a salt thereof.
1869. An agent having the structure SP-14-2 or a salt thereof.
1870. An agent having the structure SP-14-3 or a salt thereof.
1871. An agent having the structure SP-14-4 or a salt thereof.
1872. An agent having the structure SP-14-5 or a salt thereof.
1873. An agent having the structure SP-14-6 or a salt thereof.
1874. An agent having the structure SP-14-7 or a salt thereof.
1875. An agent having the structure SP-14-8 or a salt thereof.
1876. An agent having the structure SP-15-1 or a salt thereof.
1877. An agent having the structure SP-15-2 or a salt thereof.
1878. An agent having the structure SP-15-3 or a salt thereof.
1879. An agent having the structure SP-15-4 or a salt thereof.
1880. An agent having the structure SP-15-5 or a salt thereof.
1881. An agent having the structure SP-15-6 or a salt thereof.
1882. An agent having the structure SP-15-7 or a salt thereof.
1883. An agent having the structure SP-15-8 or a salt thereof.
1884. A medicament havingA structure or a salt thereof.
1885. A medicament having A structure or a salt thereof.
1886. A medicament havingA structure or a salt thereof. />
1887. The agent of any one of embodiments 1884-1886, wherein the agent has the same retention time under HPLC conditions as I-66 prepared as described in example 9, wherein the HPLC conditions can separate I-66 and I-67 prepared as described in example 9.
1888. The agent of any one of embodiments 1884-1886, wherein the agent exhibits a retention time of about 15.3 minutes under the following HPLC conditions: agilent Poroshell 120EC-C18; 4.6X100 mm; solvent a = 0.1% TFA in water; solvent B = 0.075% TFA in acetonitrile; gradient 10% B to 95% B in 30 minutes; the UV absorbance at 220nM is detected.
1889. The agent of any one of embodiments 1884-1888, wherein the agent elutes in a single peak with I-66 prepared as described in example 9 under the following HPLC conditions: agilent Poroshell 120EC-C18; 4.6X100 mm; solvent a = 0.1% TFA in water; solvent B = 0.075% TFA in acetonitrile; gradient 10% B to 95% B in 30 minutes; the UV absorbance at 220nM is detected.
1890. The agent of any one of embodiments 1884-1889, wherein the agent exhibits overlap with a peak of about 5.1-5.7 in fig. 6 under the same or equivalent conditions 1 H NMR peaks.
1891. The agent of any one of embodiments 1884-1889, wherein the agent exhibits the same about 5.1 to 5.7 as in fig. 6 under the same or equivalent conditions 1 H NMR peaks.
1892. The agent of any one of embodiments 1884 to 1889, characterized in that it is a pharmaceutical composition 1 In the H NMR spectrum, correspond to bonding to carbon atoms 1 The peak of H overlaps with the peak in fig. 6 under the same or equivalent conditions.
1893. The agent of any one of embodiments 1884 to 1889, characterized in that it 1 The H NMR spectrum overlaps with the peaks in fig. 6 under the same or comparable conditions.
1894. The agent of any one of embodiments 1884-1886, wherein the agent has the same retention time under HPLC conditions as I-67 prepared as described in example 9, wherein the HPLC conditions can separate I-66 and I-67 prepared as described in example 9.
1895. The agent of any one of embodiments 1884-1886, wherein the agent exhibits a retention time of about 16.2 minutes under the following HPLC conditions: agilent Poroshell 120EC-C18; 4.6X100 mm; solvent a = 0.1% TFA in water; solvent B = 0.075% TFA in acetonitrile; gradient 10% B to 95% B in 30 minutes; the UV absorbance at 220nM is detected.
1896. The agent of any one of embodiments 1884-1888, wherein the agent elutes in a single peak with I-67 prepared as described in example 9 under the following HPLC conditions: agilent Poroshell 120EC-C18; 4.6X100 mm; solvent a = 0.1% TFA in water; solvent B = 0.075% TFA in acetonitrile; gradient 10% B to 95% B in 30 minutes; the UV absorbance at 220nM is detected.
1897. The agent of any one of embodiments 1884-1888 and 1894-1896, wherein the agent exhibits no overlap with a peak between about 5.1 and 5.7 in fig. 6 under the same or comparable conditions 1 H NMR peaks.
1898. The agent of any one of embodiments 1884-1888 and 1894-1896, wherein the agent does not exhibit a concentration of between about 5.1 and 5.7 that is the same as in fig. 6 under identical or equivalent conditions 1 H NMR peaks.
1899. The agent of any one of embodiments 1884 to 1888 and 1894 to 1896, characterized in that it is a pharmaceutical composition 1 In the H NMR spectrum, correspond to bonding to carbon atoms 1 The peaks of H do not all overlap with the peaks in fig. 6 under the same or comparable conditions.
1900. The agent of any one of embodiments 1884 to 1889, characterized in that it 1 The H NMR spectrum does not overlap with the peaks in fig. 6 under the same or comparable conditions.
1901. A compound having the formula PA structure or a salt thereof:
N(R PA )(R a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)R PC ，
PA
wherein:
R PA is-H or an amino protecting group;
R a1 and R is a3 Each independently of the otheris-L a -R’；
R a 2 is-L aa -C(O)R PS ；
L a 、L a1 And L a2 Each independently is L;
-C(O)R PS -COOH, optionally protected or activated;
-C(O)R PC -COOH, optionally protected or activated;
each L is independently a covalent bond or an optionally substituted divalent C 1 -C 25 Aliphatic or heteroaliphatic groups having from 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-;
each-Cy-is independently an optionally substituted divalent 3-to 30-membered monocyclic, bicyclic, or polycyclic ring having 0 to 10 heteroatoms;
each R' is independently-R, -C (O) R, -CO 2 R or-SO 2 R is R; and is also provided with
Each R is independently-H or an optionally substituted group selected from: c (C) 1-30 Aliphatic, C having 1 to 10 heteroatoms 1-30 Heteroaliphatic, C 6-30 Aryl, C 6-30 Arylaliphatic, C having 1 to 10 heteroatoms 6-30 Aryl heteroaliphatics, 5 to 30 membered heteroaryl groups having 1 to 10 heteroatoms, and 3 to 30 membered heterocyclyl groups having 1 to 10 heteroatoms, or
The two R groups optionally and independently together form a covalent bond, or:
two or more R groups on the same atom optionally and independently taken together with the atom form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the atom; or alternatively
Two or more R groups on two or more atoms optionally and independently together with their intervening atoms form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the intervening atoms.
1902. A compound of embodiment 1901 wherein R a2 is-L aa -C(O)R PS Wherein L is aa Is L, and L aa comprising-N (R') -or-Cy-.
1903. The compound of any one of the preceding embodiments, wherein L a1 Is a covalent bond.
1904. The compound of any one of the preceding embodiments, wherein L a2 Is a covalent bond.
1905. The compound of any one of the preceding embodiments, wherein L aa Is an optionally substituted divalent C 1 -C 25 Aliphatic or heteroaliphatic groups having from 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 N (R') - (C (O) S) -or-C (O) O-,
wherein at least one methylene unit is replaced by-Cy-.
1906. The compound of any one of the preceding embodiments, wherein L aa is-L am1 -Cy-L am2 -, wherein L am1 And L am2 Each independently is L am Wherein each L am Independently a covalent bond or an optionally substituted divalent C 1 -C 10 An aliphatic group, wherein one or more methylene units of the aliphatic group are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-,-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
1907. The compound of any one of the preceding embodiments, wherein-L am2 -and-C (O) R PS And (5) bonding.
1908. The compound of any one of the preceding embodiments, wherein L am2 Is a covalent bond.
1909. The compound of any one of the preceding embodiments, wherein-Cy-is an optionally substituted 4-to 7-membered ring having 0 to 3 heteroatoms.
1910. A compound according to any one of the preceding embodiments, wherein-Cy-is an optionally substituted 6-to 10-membered aryl ring or an optionally substituted 5-to 10-membered heteroaryl ring having 1-5 heteroatoms.
1911. A compound according to any one of the preceding embodiments, wherein-Cy-is an optionally substituted benzene ring.
1912. The compound of any one of the preceding embodiments, wherein-Cy-is optionally substituted
1913. The compound of any one of the preceding embodiments, wherein-Cy-is
1914. The compound of any one of embodiments 1901 to 1908, wherein-Cy-is optionally substituted
1915. The compound of any one of embodiments 1901 to 1908, wherein-Cy-is
1916. The compound of any one of embodiments 1901 to 1908, wherein-Cy-is optionally substituted
1917. The compound of any one of embodiments 1901 to 1908, wherein-Cy-is
1918. The compound of any one of embodiments 1901 to 1910, wherein-Cy-is an optionally substituted 5-to 10-membered heteroaryl ring having 1 to 5 heteroatoms.
1919. The compound of any one of embodiments 1901 to 1910, wherein-Cy-is an optionally substituted 5 membered heteroaryl ring having 1 to 5 heteroatoms.
1920. The compound of any one of embodiments 1901 to 1910, wherein-Cy-is optionally substituted
1921. The compound of any one of embodiments 1901 to 1910, wherein-Cy-is
1922. The compound of any one of the preceding embodiments, wherein L aa comprising-N (R') -.
1923. The compound of embodiment 1922, wherein L aa is-L am1 -(NR’)-L am2 -, wherein L am1 And L am2 Each independently is L am Wherein each L am Independently a covalent bond or an optionally substituted divalent C 1 -C 10 An aliphatic group, wherein one or more methylene units of the aliphatic group are optionally and independently replaced by:
-C(R’) 2 -，-Cy-，-O-，-S-，-S-S-,-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 n (R') -C (O) S-, or-C (O) O-.
1924. The compound of any one of embodiments 1922 to 1923, wherein R 'and R of-N (R') -are a3 And intervening atoms together form an optionally substituted 3-to 10-membered ring having 0 to 5 heteroatoms in addition to the intervening atoms.
1925. The compound of any one of embodiments 1922 to 1924, wherein-N (R') -is bonded to two carbon atoms that do not form any double bonds with the heteroatom.
1926. The compound of any one of embodiments 1922 to 1925, wherein-L am2 -and-C (O) R PS And (5) bonding.
1927. The compound of any one of embodiments 1922 to 1926, wherein L am1 Is optionally substituted C 1-4 An alkylene group.
1928. The compound of any one of embodiments 1922 to 1926, wherein L am1 Is optionally substituted- (CH) 2 ) m-, wherein m is 1, 2, 3 or 4.
1929. The compound of any one of embodiments 1922 to 1926, wherein L am1 Is optionally substituted-CH 2 -。
1930. The compound of any one of embodiments 1922 to 1926, wherein L am1 is-CH 2 -。
1931. The compound of any one of embodiments 1922 to 1930 wherein L am2 Is optionally substituted linear C 1-2 An alkylene group.
1932. The compound of any one of embodiments 1922 to 1930 wherein L am2 Is- [ C (R') 2 ]n, where n is 1 or 2.
1933. The compound of any one of embodiments 1922 to 1930 wherein L am2 Is- [ CHR ]']n, where n is 1 or 2.
1934. The compound of any one of embodiments 1932-1933 wherein each R' is independently-H or optionally substituted C 1-6 An alkyl group.
1935. The compound of any one of embodiments 1922 to 1930 wherein L am2 Is optionally substituted-CH 2 -。
1936. The compound of any one of embodiments 1922 to 1935 wherein L am2 is-CH 2 -。
1937. The compound of any one of embodiments 1922 to 1936 wherein L aa comprising-N (R ') -, wherein R ' of-N (R ') -is-R NR Wherein R is NR Is R.
1938. The compound of any one of embodiments 1922 to 1936 wherein L aa comprising-N (R ') -, wherein R ' of-N (R ') -is-CH 2 -R NR Wherein R is NR Is R.
1939. The compound of any one of embodiments 1922 to 1936 wherein L aa comprising-N (R ') -, wherein R ' of-N (R ') -is-C (O) R NR Wherein R is NR Is R.
1940. The compound of any one of embodiments 1922 to 1936 wherein L aa comprising-N (R ') -, wherein R ' of-N (R ') -is-SO 2 R NR Wherein R is NR Is R.
1941. The compound of any one of embodiments 1937 to 1940 wherein R NR Is optionally substituted C 1-6 Aliphatic or heteroaliphatic having 1 to 4 heteroatoms.
1942. The compound of any one of embodiments 1937 to 1941 wherein R NR Is C 1-7 An alkyl group or a heteroalkyl group having 1 to 4 heteroatoms, wherein the alkyl or heteroalkyl group is optionally substituted with one or more groups independently selected from the group consisting of: halogen, C having 0 to 4 hetero atoms 5-6 An aromatic ring, an optionally substituted 3-to 10-membered cycloalkyl or a heteroalkyl ring having 1 to 4 heteroatoms.
1943. The compound of any one of embodiments 1937 to 1942 wherein R NR is-CF 3 。
1944. The compound of any one of embodiments 1937 to 1941 wherein L am2 is-C (R') 2 -or comprises-C (R') 2 -, wherein R' is a radical and L aa In (C) of (C) N (R')Taken together with the intervening atoms, form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the intervening atoms.
1945. The compound of any one of embodiments 1901 to 1905, wherein L aa Is optionally substituted C 1-4 An alkylene group.
1946. The compound of embodiment 1945 wherein L aa Is optionally substituted-CH 2 -CH 2 -。
1947. The compound of embodiment 1945 wherein L aa Is optionally substituted-CH 2 -。
1948. A compound of embodiment 1901 having the structure:
wherein:
m and n are each independently 1, 2, 3 or 4;
L RN is L;
R RN is R; and is also provided with
R a5 Is R'.
1949. A compound of embodiment 1948 wherein m is 1.
1950. The compound of any one of embodiments 1948 to 1949, wherein L RN is-CH 2 -, -CO-or-SO 2 -。
1951. The compound of any one of embodiments 1948 to 1949, wherein L RN is-CH 2 -。
1952. The compound of any one of embodiments 1948 to 1951, wherein R NR Is C 1-7 An alkyl group or a heteroalkyl group having 1 to 4 heteroatoms, wherein the alkyl or heteroalkyl group is optionally substituted with one or more groups independently selected from the group consisting of: halogen, C having 0 to 4 hetero atoms 5-6 An aromatic ring, optionally substituted 3-to 10-membered cycloalkyl orA heteroalkyl ring having 1 to 4 heteroatoms.
1953. The compound of any one of embodiments 1948 to 1952, wherein one or more R a5 Independently is-H.
1954. The compound of any one of embodiments 1948 to 1953, wherein one or more R a5 Independently optionally substituted C 1-6 An alkyl group.
1955. The compound of any one of embodiments 1948 to 1953, wherein-L RN -R RN Is R and is with R a5 And intervening atoms together form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the intervening atoms.
1956. The compound of embodiment 1951, wherein R RN Is methyl.
1957. The compound of embodiment 1951, wherein R RN is-CF 3 。
1958. The compound of any one of the preceding embodiments, wherein R a1 is-H.
1959. The compound of any one of embodiments 1901 to 1944, wherein R a1 Is optionally substituted C 1-6 An alkyl group.
1960. The compound of any one of the preceding embodiments, wherein-C (O) R PC Is a protected carboxylic acid group.
1961. The compound of any one of embodiments 1901 to 1959, wherein-C (O) R PC Is an activated carboxylic acid group.
1962. The compound of any one of embodiments 1901 to 1959, wherein-C (O) R PC is-C (O) OR'.
1963. A compound of embodiment 1962 wherein R' is-H.
1964. A compound of embodiment 1962 wherein R' is pentafluorophenyl.
1965. The compound of embodiment 1962 wherein R' is
1966. The compound of any one of the preceding embodiments, wherein-C (O) R PS is-C (O) OR'.
1967. A compound of embodiment 1966 wherein R' is-H.
1968. The compound of embodiment 1966 wherein R' is optionally substituted C 1-6 Aliphatic series.
1969. A compound of embodiment 1966 wherein R' is t-butyl.
1970. A compound of embodiment 1966 wherein R' is benzyl.
1971. A compound of embodiment 1966 wherein R' is allyl.
1972. A compound of embodiment 1901 wherein the compound hasA structure or a salt thereof, wherein ring a is an optionally substituted 3-to 7-membered saturated, partially unsaturated or aromatic ring.
1973. A compound of embodiment 1901 wherein the compound hasA structure or a salt thereof, wherein ring a is an optionally substituted 3-to 7-membered saturated, partially unsaturated or aromatic ring.
1974. A compound of any one of embodiments 1972 or 1973 wherein-C (O) OtBu is bonded to a chiral carbon atom having the R configuration.
1975. The compound of any one of embodiments 1972 or 1973 wherein-C (O) OtBu is bonded to a chiral carbon atom having an S configuration.
1976. A compound of embodiment 1901 wherein the compound hasA structure or salt thereof, wherein:
ring a is an optionally substituted 3 to 10 membered ring;
n is 0, 1 or 2;
m is 0, 1, 2 or 3.
1977. A compound of embodiment 1901 wherein the compound hasA structure or salt thereof, wherein:
ring a is an optionally substituted 3 to 10 membered ring;
n is 0, 1 or 2;
m is 0, 1, 2 or 3.
1978. The compound of any one of embodiments 1976 through 1977 wherein ring a is an optionally substituted 4-to 10-membered ring.
1979. A compound of any one of embodiments 1976 through 1978 wherein n is 1.
1980. The compound of any of embodiments 1976 through 1979 wherein ring a is attached to- (CH) 2 ) n-is bonded at the chiral carbon which is R.
The compound of any one of embodiments 1976 through 1979 wherein ring a is attached to- (CH) 2 ) n-is bonded at a chiral carbon that is S.
1982. A compound of embodiment 1901 wherein the compound hasA structure or salt thereof, wherein:
ring a is an optionally substituted 3 to 10 membered ring;
n is 0, 1 or 2;
m is 0, 1, 2 or 3.
1983. A compound of embodiment 1901 wherein the compound hasA structure or salt thereof, wherein: />
Ring a is an optionally substituted 3 to 10 membered ring;
n is 0, 1 or 2;
m is 0, 1, 2 or 3.
1984. A compound of embodiment 1901 wherein the compound hasA structure or salt thereof, wherein:
ring a is an optionally substituted 3 to 10 membered ring;
n is 0, 1 or 2;
m is 0, 1, 2 or 3.
1985. A compound of any one of embodiments 1976 through 1984 wherein n is 1.
1986. A compound of any one of embodiments 1976 through 1985 wherein m is 0.
1987. A compound of any one of embodiments 1976 through 1985 wherein m is 1, 2 or 3.
1988. A compound of any one of embodiments 1976 through 1985 wherein m is 1.
1989. The compound of any one of embodiments 1976 through 1988, wherein ring a is or comprises an optionally substituted saturated monocyclic ring.
1990. The compound of any one of embodiments 1976 through 1989 wherein ring a is or comprises an optionally substituted partially unsaturated monocyclic ring.
1991. The compound of any of embodiments 1976 through 1990 wherein ring a is or comprises an optionally substituted aromatic monocyclic ring.
1992. The compound of any one of embodiments 1982 to 1988 wherein ring a is optionally substituted phenyl.
1993. The compound of any one of embodiments 1976 through 1988 wherein ring a is an optionally substituted 5-to 6-membered heteroaryl having 1-3 heteroatoms.
1994. The compound of any one of embodiments 1976 through 1988 wherein ring a is an optionally substituted 5-to 6-membered heteroaryl having 1-3 heteroatoms, wherein at least one heteroatom is nitrogen.
1995. A compound of embodiment 1994 wherein ring a is an optionally substituted triazole ring.
1996. The compound of any one of embodiments 1976 through 1988 wherein ring a is an optionally substituted 8-to 10-membered bicyclic ring having 1 to 6 heteroatoms.
1997. The compound of any one of embodiments 1976 through 1979 wherein ring a is an optionally substituted 8-to 10-membered bicyclic aromatic ring having 1 to 6 heteroatoms, wherein each monocyclic unit is independently an optional 5-to 6-membered aromatic ring having 0 to 3 heteroatoms.
1998. The compound of any of embodiments 1993 to 1997, wherein ring a is attached to- (CH) 2 ) n-is bonded at a carbon atom.
1999. The compound of any of embodiments 1993 to 1997, wherein ring a is attached to- (CH) 2 ) n-is bonded at the nitrogen atom.
2000. The compound of any one of the preceding embodiments, wherein L aa wherein-Cy-or Ring A is optionally substituted and each substituent is independently selected from halogen, -R, -CF 3 、-N(R) 2 -CN and-OR, wherein each R is independently C optionally substituted with one OR more-F 1-6 Aliphatic series.
2001. The compound of any one of the preceding embodiments, wherein L aa Is optionally substituted, and each substituent is independently selected from halogen; c (C) 1-5 Linear, branched or cyclic alkyl; -OR, wherein R is C 1-4 Linear, branched or cyclic alkyl; fluorinated alkyl groups; -N (R) 2 -, wherein each R is independently C 1-6 Linear, branched or cyclic alkyl; or-CN.
2002. The compound of any one of the preceding embodiments, wherein R a3 is-H or optionally substituted C 1-6 Aliphatic series.
2003. The compound of any one of the preceding embodiments, wherein R a3 is-H.
2004. The chemistry of any one of embodiments 1901 to 2002 Compounds, wherein R a3 Is methyl.
2005. A compound having the following structure or a salt thereof:
wherein:
R PA is-H or an amino protecting group;
-C(O)R PS -COOH, optionally protected or activated; and is also provided with
-C(O)R PC Is an optionally protected or activated-COOH.
2006. A compound having the following structure or a salt thereof:
wherein:
R PA is-H or an amino protecting group;
-C(O)R PS -COOH, optionally protected or activated; and is also provided with
-C(O)R PC Is an optionally protected or activated-COOH.
2007. The compound of any one of the preceding embodiments, wherein R PA Is an amino protecting group suitable for peptide synthesis.
2008. The compound of any one of the preceding embodiments, wherein R PA is-C (O) -O-R.
2009. The compound of embodiment 2008, wherein R is optionally substituted
2010. The compound of any one of the preceding embodiments, wherein R PA is-Fmoc.
2011. The compound of any one of the preceding embodiments, wherein R PA is-Cbz.
2012. In the foregoing embodimentThe compound of any one of claims, wherein R PA is-Boc.
2013. The compound of any one of the preceding embodiments, wherein R PS Is with R PA Orthogonal protecting groups.
2014. The compound of any one of the preceding embodiments, wherein R PS Is with R PC Orthogonal protecting groups.
2015. The compound of any one of the preceding embodiments, wherein R PS Is compatible with peptide synthesis.
2016. The compound of any one of the preceding embodiments, wherein-C (O) R PS is-C (O) OR'.
2017. A compound of embodiment 1966 wherein R' is-H.
2018. The compound of embodiment 1966 wherein R' is optionally substituted C 1-6 Aliphatic series.
2019. A compound of embodiment 1966 wherein R' is t-butyl.
2020. A compound of embodiment 1966 wherein R' is benzyl.
2021. A compound of embodiment 1966 wherein R' is allyl.
2022. The compound of any one of embodiments 1901 to 2015, wherein-C (O) R PS is-C (O) S-L-R'.
2023. The compound of embodiment 2022, wherein L is optionally substituted-CH 2 -。
2024. The compound of embodiment 2022, wherein L is-CH 2 -。
2025. The compound of any one of embodiments 2022 to 2024, wherein R' is optionally substituted phenyl.
2026. A compound of any one of embodiments 2022 to 2024 wherein R' is 2,4, 6-trimethoxyphenyl.
2027. The compound of embodiment 2022, wherein R PS is-SH.
2028. The compound of any one of the preceding embodiments, wherein-C (O) R PC Is a protected carboxylic acid group.
2029. The compound of any one of embodiments 1901 to 2026, wherein-C (O) R PC Is an activated carboxylic acid group.
2030. The compound of any one of embodiments 1901 to 2026, wherein-C (O) R PC is-C (O) OR'.
2031. A compound of embodiment 2030 wherein R' is-H.
2032. A compound of embodiment 2030 wherein R' is pentafluorophenyl.
2033. A compound of embodiment 2030 wherein R' is
2034. A compound according to any one of the preceding embodiments wherein each heteroatom is independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon.
2035. A compound according to any one of the preceding embodiments wherein each heteroatom is independently selected from oxygen, nitrogen and sulfur.
2036. A compound, wherein the compound isOr a salt thereof.
2037. A compound, wherein the compound isOr a salt thereof. />
2038. A compound, wherein the compound isOr a salt thereof.
2039. A compound, wherein the compound isOr a salt thereof.
2040. A compound, wherein the compound isOr a salt thereof.
2041. A compound, wherein the compound is Or a salt thereof.
2042. A compound, wherein the compound isOr a salt thereof.
2043. A compound, wherein the compound isOr a salt thereof.
2044. A compound, wherein the compound isOr a salt thereof.
2045. A compound, wherein the compound isOr a salt thereof.
2046. A compound, wherein the compound isOr a salt thereof.
2047. A compound, wherein the compound isOr a salt thereof. />
2048. A compound, wherein the compound isOr a salt thereof.
2049. A compound, wherein the compound isOr a salt thereof.
2050. A compound, wherein the compound isOr a salt thereof.
2051. A compound, wherein the compound isOr a salt thereof.
2052. A compound, wherein the compound isOr a salt thereof.
2053. A compound, wherein the compound isOr a salt thereof.
2054. A compound, wherein the compound isOr a salt thereof.
2055. A compound, wherein the compound isOr a salt thereof.
2056. The compound of any one of the preceding embodiments, wherein the compound has a purity of at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%.
2057. A compound comprising a residue according to any one of the preceding embodiments.
2058. A compound comprising the residues in tables a-IV.
2059. A compound comprising a compound havingResidues of the structure or salt form thereof.
2060. A compound comprising a compound havingResidues of the structure or salt form thereof. />
2061. A compound comprising a compound havingResidues of the structure or salt form thereof.
2062. A compound comprising a compound havingResidues of the structure or salt form thereof.
2063. A compound comprising a compound havingResidues of the structure or salt form thereof.
2064. A compound comprising a compound havingResidues of the structure or salt form thereof.
2065. A compound comprising a compound havingResidues of the structure or salt form thereof.
2066. A compound comprising a compound havingResidues of the structure or salt form thereof.
2067. A compound comprising a compound havingResidues of the structure or salt form thereof.
2068. A compound comprising a compound havingResidues of the structure or salt form thereof.
2069. The compound of any one of embodiments 2057 to 2068, wherein the compound is or comprises a peptide.
2070. The compound of any one of embodiments 2057 to 2068, wherein the compound is the agent of any one of the preceding embodiments.
2071. The compound of any one of embodiments 2057 to 2068, wherein the compound is or comprises a stapled peptide.
2072. A method for preparing a compound of any one of embodiments 2057 to 2071, comprising using a compound of any one of embodiments 1901 to 2056.
2073. An agent comprising the residue of an amino acid according to any of the preceding embodiments.
2074. The agent of any one of embodiments 1 to 1900, wherein the agent comprises a residue of an amino acid of any one of the preceding embodiments.
2075. The medicament of any of the preceding embodiments, wherein each olefinic double bond in the staple-like structure is independently and optionally converted to a single bond.
2076. The medicament of any of the preceding embodiments, wherein each olefinic double bond in the staple-like structure is converted to a single bond.
2077. The agent of any one of the preceding embodiments, wherein each olefinic double bond is converted to a single bond.
2078. The agent of any one of the preceding embodiments, wherein each olefinic double bond is independently and optionally converted to-CHR' -, wherein each R is independently-H, -R, -OR, -OH, -N (R) 2 or-SR.
2079. The agent of any one of the preceding embodiments, wherein each olefinic double bond is converted to-CHR' -, wherein each R is independently-H, -R, -OR, -OH, -N (R) 2 or-SR.
2080. The agent of any one of the preceding embodiments, wherein eachThe olefinic double bonds being independently and optionally converted to optionally substituted-CH 2 -CH 2 -。
2081. The agent of any one of the preceding embodiments, wherein each olefinic double bond is converted to-CH 2 -CH 2 -。
2082. The agent of any one of the preceding embodiments, having a diastereomeric purity of about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more.
2083. The agent of any one of the preceding embodiments, having a diastereomeric purity of about 90% or greater.
2084. The agent of any one of the preceding embodiments, having a diastereomeric purity of about 95% or greater.
2085. The agent of any one of the preceding embodiments, having a diastereomeric purity of about 98% or greater.
2086. The agent of any one of the preceding embodiments having a purity of about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more.
2087. The agent of any one of the preceding embodiments having a purity of about 90% or greater.
2088. The agent of any one of the preceding embodiments having a purity of about 95% or greater.
2089. The agent of any one of the preceding embodiments having a purity of about 98% or greater.
2090. A composition comprising the agent of any one of the preceding embodiments or a salt thereof.
2091. A pharmaceutical composition comprising or delivering the agent or amino acid of any of the preceding embodiments, and a pharmaceutically acceptable carrier.
2092. A composition selected from table E2.
2093. A pharmaceutical composition comprising or delivering one or more or all of the peptide agents selected from the compositions of table E2, and a pharmaceutically acceptable carrier.
2094. A composition selected from table E3.
2095. A pharmaceutical composition comprising or delivering one or more or all of the peptide agents selected from the compositions of table E3, and a pharmaceutically acceptable carrier.
2096. The composition of any of the preceding embodiments, comprising a medicament comprising one or more staple-like structures, each independently comprising one or more olefinic double bonds.
2097. The composition of any of the preceding embodiments, wherein the ratio of the two stereoisomers of the olefinic double bond in the staple-like structure is about 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 30:1, 40:1, 50:1 or higher.
2098. The composition of embodiment 2097, wherein the ratio is about 5:1 or greater.
2099. The composition of embodiment 2097, wherein the ratio is about 10:1 or greater.
2100. The composition of embodiment 2097, wherein the ratio is about 20:1 or greater.
2101. The composition of embodiment 2097, wherein the ratio is about 50:1 or greater.
2102. The composition of any of the preceding embodiments, wherein each ratio of the two stereoisomers of each olefinic double bond in each staple-like structure is independently about 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 30:1, 40:1, 50:1 or higher.
2103. The composition of embodiment 2102, wherein each ratio is independently about 5:1 or higher.
2104. The composition of embodiment 2102, wherein each ratio is independently about 10:1 or higher.
2105. The composition of embodiment 2102, wherein each ratio is independently about 20:1 or higher.
2106. The composition of embodiment 2102, wherein each ratio is independently about 50:1 or higher.
2107. The composition of any of the preceding embodiments, wherein the selectivity is biased towards the E configuration.
2108. The composition of any of the preceding embodiments, wherein the selectivity is biased toward the Z configuration.
2109. A method for preparing the agent or composition of any of the preceding embodiments, comprising incorporating the residue of the amino acid of any of the preceding embodiments.
2110. The method of embodiment 2109 comprising preparing a compound comprising one or more terminal olefin-containing amino acid residues, wherein one or more or all of such amino acid residues are not stapled.
2111. A method, which comprises
a) Preparing a first compound comprising two moieties, each independently comprising an olefinic double bond;
b) Stapling one portion of the olefinic double bond with another portion of the olefinic double bond by olefin metathesis of the two portions to form a first formed staple-like structure, thereby providing a second compound;
c) Adding one or more additional moieties to the second compound to provide a third compound comprising two moieties, each independently comprising an olefinic double bond; and
d) The two portions of the third compound are stapled by olefin metathesis of one portion of the olefin double bond with the other portion of the olefin double bond to form a second formed staple-like structure, thereby providing a fourth compound.
2112. The method of embodiment 2111, wherein each moiety is independently an amino acid residue comprising a terminal olefin of any of the preceding embodiments.
2113. The method of any of the preceding embodiments, wherein there are two olefinic double bonds in one portion of the first compound.
2114. The method of any of the preceding embodiments, wherein the moiety in the first compound is an amino acid residue comprising two olefinic double bonds.
2115. The method of any one of the preceding embodiments, wherein one moiety in the first compound is B5.
2116. The method of any of the embodiments, wherein the two moieties in the first compound are independently X 4 And X 11 。
2117. The method of any of the preceding embodiments, wherein the first formed staple-like structure is an (i, i+7) staple-like structure.
2118. The method of any of the preceding embodiments, wherein the first compound comprises-X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 -。
2119. The method of any of the preceding embodiments, wherein the first compound comprises-X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 -。
2120. The method of any of the preceding embodiments, wherein the first compound comprises a staple-like structure.
2121. The method of embodiment 2120, wherein the staple-like structure is an (i, i+4) staple-like structure.
2122. The method of embodiment 2120, wherein the staple-like structure is at X 10 And X is 14 Between them.
2123. The method of any of the preceding embodiments, wherein the olefinic double bond in the third compound is present in the first compound.
2124. The method of any one of the preceding embodiments, wherein one and only one amino acid residue comprises an olefinic double bond, which is added to the second compound.
2125. The method of any one of the preceding embodiments, wherein the third compound is-X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 -or comprise-X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 -。
2126. The method of any one of the preceding embodiments, wherein the third compound is-X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 -or comprise-X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 -。
2127. The method of any of the preceding embodiments, wherein the first formed staple-like structure and the second formed staple-like structure are bonded to the same amino acid residue.
2128. The method of any of the preceding embodiments, wherein the first formed staple-like structure and the second formed staple-like structure are bonded to the same atom.
2129. The method of any of the preceding embodiments, wherein the second formed staple-like structure is an (i, i+2), (i, i+3), or (i, i+4) staple-like structure.
2130. The method of any one of the preceding embodiments, wherein the two moieties in the third compound are independently X 1 And X 4 。
2131. The method of any of the preceding embodiments, wherein the first formed staple-like structure is selectively formed with E.
2132. The method of any of the preceding embodiments, wherein the second formed staple-like structure is formed with Z-selectivity.
2133. The method of any one of embodiments 2111 to 2132, wherein the agent of any one of the preceding embodiments is prepared.
2134. The method of any one of the preceding embodiments, comprising preparing a compound having an amino acid sequence of table E2 or table E3 but one or more or all of the amino acid residues comprising a terminal olefin that are not stapled.
2135. The method of any one of embodiments 2109 to 2134, comprising stapling two or more amino acid residues, each independently comprising one or more olefins, to form one or more staple-like structures, each independently comprising a carbon-carbon double bond.
2136. The method of embodiment 2135, wherein the stapling is performed by olefin metathesis of terminal olefins.
2137. The method of any one of embodiments 2109 to 2136, wherein the double bond in the staple-like structure is formed with a stereoselectivity of about 1.1:1, 1.2:1, 1.5:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1 or more.
2138. The method of embodiment 2137, wherein the selectivity is about 1.5:1 or greater.
2139. The method of embodiment 2137, wherein the selectivity is about 2:1 or greater.
2140. The method of embodiment 2137, wherein the selectivity is about 3:1 or greater.
2141. The method of embodiment 2137, wherein the selectivity is about 4:1 or greater.
2142. The method of embodiment 2137, wherein the selectivity is about 9:1 or greater.
2143. The method of embodiment 2137, wherein the selectivity is about 10:1 or greater.
2144. The method of embodiments 2137-2143, wherein the staple-like structure is a first formed staple-like structure.
2145. The method of any one of embodiments 2109 to 2144, wherein each double bond in each staple-like structure is independently formed with a stereoselectivity of about 1.1:1, 1.2:1, 1.5:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1 or more.
2146. The method of embodiment 2145, wherein the selectivity is independently about 1.5:1 or higher.
2147. The method of embodiment 2145, wherein the selectivity is independently about 2:1 or higher.
2148. The method of embodiment 2145, wherein the selectivity is independently about 3:1 or higher.
2149. The method of embodiment 2145, wherein the selectivity is independently about 4:1 or higher.
2150. The method of embodiment 2145, wherein the selectivity is independently about 9:1 or higher.
2151. The method of embodiment 2145, wherein the selectivity is independently about 10:1 or higher.
2152. The method of any one of embodiments 2137 to 2151, wherein selectivity is biased towards the E isomer.
2153. The method of any one of embodiments 2137 to 2151, wherein selectivity is biased towards the Z isomer.
2154. The method of any one of embodiments 2109 to 2153, comprising purifying the composition to enrich for one or more E/Z stereoisomers.
2155. The method of embodiment 2154, wherein one configuration of olefinic double bonds in the staple-like structure is enriched.
2156. The method of embodiment 2155, wherein the ratio after enrichment is about 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 30:1, 40:1, 50:1, or higher.
2157. The method of embodiment 2156, wherein the ratio is about 5:1 or higher.
2158. The method of embodiment 2156, wherein the ratio is about 10:1 or higher.
2159. The method of embodiment 2156, wherein the ratio is about 20:1 or higher.
2160. The method of embodiment 2156, wherein the ratio is about 50:1 or higher.
2161. The method of embodiment 2154, wherein the configuration of each olefinic double bond in each staple-like structure is enriched independently.
2162. The method of embodiment 2161, wherein the ratio of each olefinic double bond after enrichment is independently about 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 30:1, 40:1, 50:1 or higher.
2163. The method of embodiment 2162, wherein each ratio is independently about 5:1 or higher.
2164. The method of embodiment 2162, wherein each ratio is independently about 10:1 or higher.
2165. The method of embodiment 2162, wherein each ratio is independently about 20:1 or higher.
2166. The method of embodiment 2162, wherein each ratio is independently about 50:1 or higher.
2167. The method of any of embodiments 2154 through 2166, wherein the selectivity is biased towards the E configuration.
2168. The method of any of embodiments 2154 to 2167, wherein the selectivity is biased towards the Z configuration.
2169. The method of any of the preceding embodiments, wherein the chiral center is formed with a stereoselectivity of about or at least about: 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99%.
2170. The method of any of the preceding embodiments, wherein the chiral center is formed with a stereoselectivity of about 80% or at least about 80% or more.
2171. The method of any of the preceding embodiments, wherein the chiral center is formed with a stereoselectivity of about 85% or at least about 85% or more.
2172. The method of any of the preceding embodiments, wherein the chiral center is formed with a stereoselectivity of about 90% or at least about 90% or more.
2173. The method of any of the preceding embodiments, wherein the chiral center is formed with a stereoselectivity of about 95% or at least about 95% or more.
2174. The method of any one of embodiments 2169-2173, wherein the chiral center is bonded to two staple-like structures.
2175. The method of any one of embodiments 2109 to 2174, comprising modifying double bonds in the staple-like structure.
2176. The method of any one of embodiments 2109 to 2174, comprising hydrogenating double bonds in the staple-like structure.
2177. The method of any one of embodiments 2109 to 2174, comprising hydrogenating each carbon-carbon double bond in each staple-like structure.
2178. The method of any of the preceding embodiments, comprising purifying the composition by chromatography and providing one or more compositions based on peaks observed during purification.
2179. The method of any of the preceding embodiments, comprising purifying the composition by liquid chromatography and providing one or more compositions based on peaks observed during purification.
2180. The method of any one of embodiments 2178 to 2179, wherein the chromatographic purification uses the same or similar conditions as those described in table E2 or table E3 in isolating peaks with correct mass.
2181. The method of any one of embodiments 2178 to 2180, wherein the chromatographic purification uses the same or similar conditions as those described in table E2 or table E3 in terms of the elution order of peaks with correct mass.
2182. The method of any of the preceding embodiments, comprising collecting as a product composition a first peak having the correct mass.
2183. The method of any of the preceding embodiments, comprising collecting a second peak having the correct mass as a product composition.
2184. The method of any of the preceding embodiments, comprising collecting a third peak having the correct mass as a product composition.
2185. The method of any of the preceding embodiments, comprising collecting a fourth peak having the correct mass as a product composition.
2186. The method of any of the preceding embodiments, comprising collecting each peak having the correct mass as a product composition.
2187. The method of any of the preceding embodiments, wherein the peak area of the product composition is about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more of the total peak area of all peaks having the correct mass.
2188. The method of embodiment 2187, wherein the percentage is 10% or greater.
2189. The method of embodiment 2187, wherein the percentage is 20% or greater.
2190. The method of embodiment 2187, wherein the percentage is 50% or greater.
2191. The method of embodiment 2187, wherein the percentage is 60% or greater.
2192. The method of any of the preceding embodiments, wherein the peak area of each product composition is about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more of the total peak area of all peaks having the correct mass.
2193. The method of embodiment 2192, wherein each percentage is independently 10% or greater.
2194. The method of embodiment 2192, wherein each percentage is independently 20% or higher.
2195. The method of embodiment 2192, wherein each percentage is independently 50% or greater.
2196. The method of embodiment 2192, wherein each percentage is independently 60% or higher.
2197. The method of any one of embodiments 2187 to 2196, wherein the peak is from MS detection.
2198. The method of any one of embodiments 2187 to 2197, wherein the peak is from UV detection.
2199. The method of any one of embodiments 2187 to 2197, wherein the peak is from UV detection at 220 nm.
2200. A composition produced by the method of any one of the preceding embodiments.
2201. A pharmaceutical composition comprising or delivering the composition of embodiment 2200 and a pharmaceutically acceptable carrier.
2202. A method for modulating β -catenin interaction with a partner in a system comprising contacting β -catenin with the agent or composition of any of the preceding embodiments.
2203. A method for modulating β -catenin interaction with a partner in a system comprising administering or delivering to the system the agent or composition of any of the preceding embodiments.
2204. The method of any one of embodiments 2202 to 2203, wherein said partner is TCF7, LEF1, TCF7L2, axinl, axin2, or APC.
2205. A method for modulating TCF- β -catenin interaction in a system comprising contacting β -catenin with the agent or composition of any of the preceding embodiments.
2206. A method for modulating TCF- β -catenin interaction in a system comprising administering or delivering to the system an agent or composition according to any of the preceding embodiments.
2207. A method for inhibiting β -catenin dependent cell proliferation comprising administering or delivering to the system the agent or composition of any of the preceding embodiments.
2208. A method for modulating the WNT/β -catenin pathway in a system, comprising administering or delivering to the system the agent or composition of any one of the preceding embodiments, wherein expression of a nucleic acid is modulated.
2209. A method comprising administering or delivering the agent or composition of any of the preceding embodiments to the system, wherein the level of nucleic acid transcripts and/or products thereof is modulated.
2210. A method comprising administering or delivering the agent or composition of any of the preceding embodiments to the system, wherein expression of the nucleic acid is modulated.
2211. The method of any one of embodiments 2208 to 2210, wherein the nucleic acid is or comprises a gene.
2212. The method of any one of embodiments 2208 to 2211, wherein the nucleic acid is selected from the gene set bcat_gds748_up or table GS1.
2213. The method of any one of embodiments 2208 to 2212, wherein the nucleic acid is selected from the gene set bcat.100_up.v1_up or table GS2.
2214. The method of any one of embodiments 2208 to 2213, wherein the nucleic acid is selected from the gene set hallmark_wnt_beta_catenin_signature or table GS3.
2215. The method of any one of embodiments 2208 TO 2214, wherein the nucleic acid is selected from the gene set rashi_response_to_ionjradiation_1 or table GS4.
2216. The method of any one of embodiments 2208 to 2215, wherein the nucleic acid is selected from the gene set REACTOME_RRNA_PROCESSING or table GS5.
2217. The method of any one of embodiments 2208 to 2216, wherein the nucleic acid is selected from the gene set hallmark_myc_target_v1 or table GS6.
2218. The method of any one of embodiments 2208 to 2217, wherein the nucleic acid is selected from the gene set hallmark_myc_target_v2 or table GS7.
2219. The method of any one of embodiments 2208 to 2218, wherein the nucleic acid is selected from the gene set hallmark_oxidation_PHOSPHORYLATION or table GS8.
2220. The method of any one of embodiments 2208 to 2219, wherein the nucleic acid is selected from the gene set hallmark_e2f_target or table GS9.
2221. The method of any one of embodiments 2208 to 2220, wherein the nucleic acid is selected from the gene set hallmark_tnfa_signaling_via_nfkb or table GS10.
2222. The method of any one of embodiments 2208 to 2221, wherein the nucleic acid is SP5.
2223. The method of any one of embodiments 2208 to 2222, wherein the nucleic acid is CCND2.
2224. The method of any one of embodiments 2208 to 2223, wherein the nucleic acid is WNT5B.
2225. The method of any one of embodiments 2208 to 2224, wherein the nucleic acid is AXIN2.
2226. The method of any one of embodiments 2208 to 2225, wherein the nucleic acid is NKD1.
2227. The method of any one of embodiments 2208 to 2226, wherein the nucleic acid is WNT6.
2228. The method of any one of embodiments 2208 to 2227, wherein the nucleic acid is DKK1.
2229. The method of any one of embodiments 2208 to 2228, wherein the nucleic acid is DKK4.
2230. The method of any one of embodiments 2208 to 2229, wherein the expression of the nucleic acid is reduced.
2231. The method of any one of embodiments 2208 to 2230, wherein bcat_gds748_up is negatively enriched.
2232. The method of any of embodiments 2208 to 2231, wherein bcat.100_up.v1_up is negatively enriched.
2233. The method of any of embodiments 2208 to 2232, wherein hallmark_wnt_beta_catenin_signaling is negatively enriched.
2234. The method of any one of embodiments 2208 TO 2233, wherein rashi_response_to_ionjradial_1 is negatively enriched.
2235. The method of any one of embodiments 2208 to 2234, wherein REACTOME_RRNA_PROCESSING is negatively enriched.
2236. The method of any one of embodiments 2208 to 2235, wherein hallmark_myc_target_v1 is negatively enriched.
2237. The method of any one of embodiments 2208 to 2236, wherein hallmark_myc_target_v2 is negatively enriched.
2238. The method of any of embodiments 2208 to 2237, wherein hallmark_oxidation_PHOSPHORYLATION is negatively enriched.
2239. The method of any one of embodiments 2208 to 2238, wherein hallmark_e2f_target is negatively enriched.
2240. The method of any one of embodiments 2208 to 2239, wherein hallmark_tnfa_signaling_via_nfkb is negatively enriched.
2241. The method of any one of embodiments 2208 to 2240, wherein the expression of the nucleic acid is reduced.
2242. The method of any one of embodiments 2208 to 2241, wherein the level of the transcript and/or product thereof is reduced.
2243. The method of any one of embodiments 2208 to 2242, wherein expression of the nucleic acid is increased.
2244. The method of any one of embodiments 2208 to 2243, wherein the level of the nucleic acid transcript or product thereof is increased.
2245. The method of any one of embodiments 2243 to 2244, wherein said nucleic acid is or comprises a CXCL12 gene.
2246. The method of any one of embodiments 2208 to 2245, wherein one or more gene sets are independently being enriched.
2247. The method of any one of embodiments 2202 to 2246, wherein the system is an in vitro system.
2248. The method of any one of embodiments 2202 to 2246, wherein the system is an in vivo system.
2249. The method of any one of embodiments 2202 to 2246, wherein the system is or comprises a sample.
2250. The method of any one of embodiments 2202 to 2249, wherein the system is or comprises a cell, tissue or organ.
2251. The method of any one of embodiments 2202 to 2250, wherein the system is or comprises a cancer cell.
2252. The method of any one of embodiments 2202 to 2251, wherein the system is or comprises colorectal cancer cells.
2253. The method of any one of embodiments 2202 to 2253, wherein the system is or comprises COLO320DM cells.
2254. The method of any one of embodiments 2202 to 2253, wherein the system is or comprises a tumor.
2255. The method of any one of embodiments 2202 to 2254, wherein the system is a subject.
2256. A method for treating or preventing a condition, disorder or disease associated with β -catenin in a subject comprising administering or delivering to the subject an effective amount of the agent or composition of any of the preceding embodiments.
2257. A method for treating cancer in a subject comprising administering or delivering to the subject an effective amount of the agent or composition of any of the preceding embodiments.
2258. A method for treating or preventing a condition, disorder or disease associated with β -catenin interaction with a partner in a subject, comprising administering or delivering to the subject an effective amount of the agent or composition of any of the preceding embodiments.
2259. The method of embodiment 2258, wherein said partner is TCF7, LEF1, TCF7L2, axin1, axin2, or APC.
2260. A method for treating or preventing a condition, disorder or disease associated with TCF- β -catenin interaction in a subject comprising administering or delivering to the subject an effective amount of the agent or composition of any of the preceding embodiments.
2261. The method of any one of the preceding embodiments, wherein the condition, disorder or disease is melanoma.
2262. The method of any of the preceding embodiments, comprising administering or delivering a second therapeutic agent to the subject.
2263. The method of any of the preceding embodiments, comprising administering or delivering a second treatment to the subject.
2264. The method of embodiment 2262 or 2263, wherein the second therapeutic agent or treatment is administered prior to the agent of any of the preceding embodiments.
2265. The method of embodiment 2262 or 2263, wherein the second therapeutic agent or treatment is administered about or no more than about 1, 2, 3, 4, 5, 6, or 7 days, or 1, 2, 3, or weeks, or 1, 2, 3, 4, 5, or 6 months prior to the agent of any of the preceding embodiments.
2266. The method of embodiment 2262 or 2263, wherein the second therapeutic agent or treatment is administered simultaneously with the agent of any of the preceding embodiments.
2267. The method of embodiment 2262 or 2263, wherein the second therapeutic agent or treatment is administered after the agent of any of the preceding embodiments.
2268. The method of embodiment 2262 or 2263, wherein the second therapeutic agent or treatment is administered about or no more than about 1, 2, 3, 4, 5, 6, or 7 days, or 1, 2, 3, or weeks, or 1, 2, 3, 4, 5, or 6 months after the agent of any of the preceding embodiments.
2269. The method of any one of the preceding embodiments, wherein the subject is exposed to a second therapeutic agent or treatment and the agent of any one of the preceding embodiments.
2270. The method of any one of the preceding embodiments, wherein the subject is exposed to the therapeutic effect of the second therapeutic agent or treatment and the therapeutic effect of the agent of any one of the preceding embodiments.
2271. The method of any one of the preceding embodiments, wherein the second therapeutic agent is or comprises a chemotherapeutic agent.
2272. The method of any one of the preceding embodiments, wherein the second therapeutic agent is or comprises a hormonal therapeutic agent.
2273. The method of any one of the preceding embodiments, wherein the second therapeutic agent is or comprises an immunotherapeutic agent.
2274. The method of any one of the preceding embodiments, wherein the second therapeutic agent is or comprises a checkpoint inhibitor.
2275. The method of any one of the preceding embodiments, wherein the second therapeutic agent is or comprises an antibody.
2276. The method of any one of the preceding embodiments, wherein the second therapeutic agent is a CTLA-4, PD-1, or PD-L1 inhibitor, or comprises a CTLA-4, PD-1, or PD-L1 inhibitor.
2277. The method of any one of the preceding embodiments, wherein the second therapeutic agent is or comprises a cell.
2278. The method of any one of the preceding embodiments, wherein the second therapeutic agent reduces one or more side effects of the agent or composition of any one of the preceding embodiments.
2279. The method of any one of the preceding embodiments, wherein the agent or composition reduces one or more side effects of the second therapeutic agent.
2280. The method of any one of the preceding embodiments, wherein the second treatment is or comprises surgery.
2281. The method of any one of the preceding embodiments, wherein the second treatment is or comprises chemotherapy.
2282. The method of any of the preceding embodiments, wherein the second treatment is or comprises radiation therapy.
2283. The method of any one of the preceding embodiments, wherein the second treatment is or comprises hormonal therapy.
2284. The method of any one of the preceding embodiments, wherein the second treatment is or comprises a stem cell or bone marrow transplant.
2285. The method of any one of the preceding embodiments, wherein the second treatment is or comprises an immunotherapy.
2286. The method of any one of the preceding embodiments, wherein the second treatment is or comprises a T cell treatment.
2287. The method of any one of the preceding embodiments, wherein the second treatment is CAR T cell treatment or comprises CART cell treatment.
2288. The method of any one of the preceding embodiments, wherein the second treatment is or comprises administering to the subject a population of immune cells.
2289. The method of any one of the preceding embodiments, wherein the agent or composition reduces one or more side effects of the second treatment.
2290. The method of any of the preceding embodiments, wherein the unit dose of the second therapeutic or therapeutic agent is reduced compared to when it is administered alone.
2291. The method of any of the preceding embodiments, wherein the total dose of the second treatment or therapeutic agent is reduced compared to when it is administered alone.
2292. The method of any of the preceding embodiments, wherein the unit dose of the agent or composition of any of the preceding embodiments is reduced as compared to when administered alone.
2293. The method of any of the preceding embodiments, wherein the total dose of the agent or composition of any of the preceding embodiments is reduced as compared to when administered alone.
2294. The method of any of the preceding embodiments, wherein the combination therapy provides greater efficacy than administration or delivery of the agent or composition alone.
2295. The method of any of the preceding embodiments, wherein the combination therapy provides greater efficacy than when the second therapeutic agent or therapy is administered or delivered alone.
2296. The method of any one of the preceding embodiments, comprising assessing expression of the nucleic acid.
2297. The method of any one of the preceding embodiments, wherein expression of the nucleic acid is modulated.
2298. The method of any one of the preceding embodiments, comprising assessing the level of a nucleic acid transcript and/or product thereof.
2299. The method of any one of the preceding embodiments, wherein the level of the nucleic acid transcript and/or product thereof is modulated.
2300. The method of any one of the preceding embodiments, comprising collecting a sample from a subject and assessing expression of nucleic acids in the sample.
2301. The method of any one of the preceding embodiments, comprising collecting a sample from a subject, wherein expression of nucleic acid in the sample is modulated.
2302. The method of any one of the preceding embodiments, comprising collecting a sample from the system and assessing expression of nucleic acid in the sample.
2303. The method of any one of the preceding embodiments, comprising collecting a sample from the system, wherein expression of nucleic acid in the sample is modulated.
2304. The method of any one of the preceding embodiments, comprising collecting a sample from a subject and assessing the level of nucleic acid transcripts and/or products thereof in the sample.
2305. The method of any one of the preceding embodiments, comprising collecting a sample from a subject, and the level of nucleic acid transcripts and/or products thereof in the sample is modulated.
2306. The method of any one of the preceding embodiments, comprising collecting a sample from the system and assessing the level of nucleic acid transcripts and/or products thereof in the sample.
2307. The method of any one of the preceding embodiments, comprising collecting a sample from the system, and the level of nucleic acid transcripts and/or products thereof in the sample is modulated.
2308. The method of any one of embodiments 2296 to 2307, wherein the sample is or comprises a cell, tissue or organ.
2309. The method of any one of embodiments 2296 to 2308, wherein the sample is or comprises a cancer cell.
2310. The method of any one of embodiments 2296 to 2309, wherein the sample is or comprises colorectal cancer cells.
2311. The method of any one of embodiments 2296 to 2310, wherein the sample is or comprises COLO320DM cells.
2312. The method of any one of embodiments 2296 to 2311, wherein the sample comprises cells from a tumor.
2313. The method of any one of embodiments 2296 to 2312, wherein the sample comprises tissue from a tumor.
2314. The method of any one of embodiments 2296 to 2313, wherein the sample is or comprises a tumor.
2315. The method of any one of embodiments 2296 to 2311, wherein the sample is from a tumor.
2316. The method of any one of embodiments 2296 to 2315, wherein the sample is from a biopsy.
2317. The method of any one of embodiments 2296 to 2316, wherein the sample is collected after one or more applications or deliveries.
2318. The method of any one of embodiments 2296 to 2317, wherein evaluation is performed after one or more administrations or deliveries.
2319. The method of any one of embodiments 2296 to 2318, wherein the nucleic acid is or comprises a gene.
2320. The method of any one of embodiments 2296 to 2319, wherein the nucleic acid is selected from the gene set bcat_gds748_up or table GS1.
2321. The method of any one of embodiments 2296 to 2320, wherein the nucleic acid is selected from the gene set bcat.100_up.v1_up or table GS2.
2322. The method of any one of embodiments 2296 to 2321, wherein the nucleic acid is selected from the gene set hallmark_wnt_beta_catenin_signature or table GS3.
2323. The method of any one of embodiments 2296 TO 2322, wherein the nucleic acid is selected from the gene set rashi_response_to_ionizingjradiation_1 or table GS4.
2324. The method of any one of embodiments 2296 to 2323, wherein the nucleic acid is selected from the gene set read_rrna_process or table GS5.
2325. The method of any one of embodiments 2296 to 2324, wherein the nucleic acid is selected from the gene set hallmark_myc_target_v1 or table GS6.
2326. The method of any one of embodiments 2296 to 2325, wherein the nucleic acid is selected from the gene set hallmark_myc_target_v2 or table GS7.
2327. The method of any one of embodiments 2296 to 2326, wherein the nucleic acid is selected from the gene set hallmark_oxidation_PHOSPHORYLATION or table GS8.
2328. The method of any one of embodiments 2296 to 2327, wherein the nucleic acid is selected from the gene set hallmark_e2f_target or table GS9.
2329. The method of any one of embodiments 2296 to 2328, wherein the nucleic acid is selected from the gene set hallmark_tnfa_signaling_via_nfkb or table GS10.
2330. The method of any one of embodiments 2296 to 2329, wherein the nucleic acid is CCND2.
2331. The method of any one of embodiments 2296 to 2330, wherein the nucleic acid is WNT5B.
2332. The method of any one of embodiments 2296 to 2331, wherein the nucleic acid is AXIN2.
2333. The method of any one of embodiments 2296 to 2332, wherein the nucleic acid is NKD1.
2334. The method of any one of embodiments 2296 to 2333, wherein the nucleic acid is WNT6.
2335. The method of any one of embodiments 2296 to 2334, wherein the nucleic acid is DKK1.
2336. The method of any one of embodiments 2296 to 2335, wherein the nucleic acid is DKK4.
2337. The method of any one of embodiments 2296 to 2336, wherein the expression of the nucleic acid is reduced.
2338. The method of any one of embodiments 2296 to 2337, wherein bcat_gds748_up is negatively enriched.
2339. The method of any one of embodiments 2296 to 2338, wherein bcat.100_up.v1_up is negatively enriched.
2340. The method of any one of embodiments 2296 to 2339, wherein hallmark_wnt_beta_catenin_signaling is negatively enriched.
2341. The method of any one of embodiments 2296 TO 2340, wherein rashi_response_to_ioniz_radiation_1 is negatively enriched.
2342. The method of any one of embodiments 2296 to 2341, wherein REACTOME_RRNA_PROCESSING is negatively enriched.
2343. The method of any one of embodiments 2296 to 2342, wherein hallmark_myc_target_v1 is negatively enriched.
2344. The method of any one of embodiments 2296 to 2343, wherein hallmark_myc_target_v2 is negatively enriched.
2345. The method of any of embodiments 2296 to 2344, wherein hallmark_oxidation_PHOSPHORYLATION is negatively enriched.
2346. The method of any one of embodiments 2296 to 2345, wherein hallmark_e2f_target is negatively enriched.
2347. The method of any one of embodiments 2296 to 2346, wherein hallmark_tnfa_signaling_via_nfkb is negatively enriched.
2348. The method of any one of embodiments 2296 to 2347, wherein the expression of the nucleic acid is reduced.
2349. The method of any one of embodiments 2296 to 2348, wherein the level of the transcript and/or product thereof is reduced.
2350. The method of any one of embodiments 2296 to 2349, wherein expression of the nucleic acid is increased.
2351. The method of any one of embodiments 2296 to 2350, wherein the level of a nucleic acid transcript or product thereof is increased.
2352. The method of any one of embodiments 2350 to 2351, wherein the nucleic acid is or comprises a CXCL12 gene.
2353. The method of any one of embodiments 2296 to 2352, wherein one or more gene sets are independently positive for enrichment.
2354. The method of any one of embodiments 2296 to 2353, wherein administration or delivery is continued one or more times after evaluation.
2355. The method of any one of embodiments 2296 to 2354, comprising evaluating and continuing administration or delivery.
2356. The method of any one of embodiments 2296 to 2355, wherein administration or delivery is adjusted after evaluation.
2357. The method of any one of embodiments 2296 to 2356, comprising evaluating and adjusting the administration or delivery.
2358. The method of any one of embodiments 2296 to 2319, wherein administration or delivery is discontinued after assessment.
2359. The method of any one of embodiments 2296 to 2318 and 2358, comprising evaluating and suspending the administration or delivery.
2360. The method of any one of embodiments 2356 to 2359, wherein the expression of SP5 is maintained about the same or is increased.
2361. The method of any one of embodiments 2356 to 2360, wherein expression of CCND2 is maintained about the same or increased.
2362. The method of any one of embodiments 2356 to 2361, wherein the expression of WNT5B remains about the same or is increased.
2363. The method of any one of embodiments 2356 to 2362, wherein the expression of AXIN2 is maintained about the same or is increased.
2364. The method of any one of embodiments 2356 to 2363, wherein the expression of NKD1 is maintained about the same or increased.
2365. The method of any one of embodiments 2356 to 2364, wherein the expression of WNT6 remains about the same or is increased.
2366. The method of any one of embodiments 2356-2365, wherein the expression of DKK1 is maintained about the same or increased.
2367. The method of any one of embodiments 2356-2366, wherein the expression of DKK4 is maintained about the same or increased.
2368. The method of any one of embodiments 2356 to 2367, wherein the expression of bcat_gds748_up or one or more nucleic acids in table GS1 independently remains about the same or is increased.
2369. The method of any one of embodiments 2356 to 2368, wherein the expression of one or more nucleic acids in bcat.100_up.v1_up or table GS2 independently remains about the same or is increased.
2370. The method of any one of embodiments 2356 to 2369, wherein the expression of one or more nucleic acids of hallmark_wnt_beta_catenin_signaling or table GS3 independently remains about the same or is increased.
2371. The method of any one of embodiments 2356 TO 2370, wherein the expression of one or more nucleic acids of rashi_response_to_ionjradiation_1 or table GS4 independently remains about the same or is increased.
2372. The method of any one of embodiments 2356 to 2371, wherein the expression of one or more nucleic acids in REACTOME_RRNA_PROCESSING or Table GS5 independently remains about the same or is increased.
2373. The method of any one of embodiments 2356 to 2372, wherein the expression of one or more nucleic acids in hallmark_myc_target_v1 or table GS6 independently remains about the same or is increased.
2374. The method of any one of embodiments 2356 to 2373, wherein the expression of one or more nucleic acids in hallmark_myc_target_v2 or table GS7 independently remains about the same or is increased.
2375. The method of any one of embodiments 2356 to 2374, wherein the expression of one or more nucleic acids of hallmark_oxidation_PHOSPHORYLATION or table GS8 independently remains about the same or is increased.
2376. The method of any one of embodiments 2356 to 2375, wherein the expression of one or more nucleic acids in hallmark_e2f_target or table GS9 independently remains about the same or is increased.
2377. The method of any one of embodiments 2356 to 2376, wherein the expression of one or more nucleic acids in hallmark_tnfa_signaling_via_nfkb or table GS10 independently remains about the same or is increased.
2378. The method of any one of embodiments 2356-2377, wherein the expression of CXCL12 independently remains about the same or is reduced.
2379. The method of any one of embodiments 2356 to 2378, wherein bcat_gds748_up is not enriched or is being enriched.
2380. The method of any one of embodiments 2356 to 2379, wherein bcat.100_up.v1_up is not enriched or is being enriched.
2381. The method of any of embodiments 2356 to 2380, wherein hallmark_wnt_beta_catenin_signaling is not enriched or is being enriched.
2382. The method of any one of embodiments 2356 TO 2381, wherein rashi_response_to_ioniz_radiation_1 is not enriched or is being enriched.
2383. The method of any one of embodiments 2356 to 2382, wherein REACTOME_RRNA_PROCESSING is not enriched or is being enriched.
2384. The method of any one of embodiments 2356 to 2383, wherein hallmark_myc_target_v1 is not enriched or is being enriched.
2385. The method of any one of embodiments 2356 to 2384, wherein hallmark_myc_target_v2 is not enriched or is being enriched.
2386. The method of any of embodiments 2356 to 2385, wherein hallmark_oxidation_PHOSPHORYLATION is not enriched or is being enriched.
2387. The method of any one of embodiments 2356 to 2386, wherein hallmark_e2f_target is not enriched or is being enriched.
2388. The method of any one of embodiments 2356 to 2387, wherein hallmark_tnfa_signaling_via_nfkb is not enriched or is being enriched.
2389. The method of any of the preceding embodiments, wherein the comparison (e.g., decreasing, increasing, enriching, negative enriching, positive enriching, etc.) is a reference assessment prior to any administration or delivery.
2390. The method of any of the preceding embodiments, wherein the comparison (e.g., decreasing, increasing, enriching, negative enriching, positive enriching, etc.) is a reference assessment of the sample prior to any administration or delivery.
2391. The method of any of the preceding embodiments, wherein the comparison (e.g., decreasing, increasing, enriching, negative enriching, positive enriching, etc.) is a reference assessment at or during administration or delivery.
2392. The method of any of the preceding embodiments, wherein the comparison (e.g., decreasing, increasing, enriching, negative enriching, positive enriching, etc.) is a reference assessment of the collected sample at or during administration or delivery.
2393. The method of any of the preceding embodiments, wherein the comparison (e.g., decreasing, increasing, enriching, negative enriching, positive enriching, etc.) is a reference assessment after early administration or delivery.
2394. The method of any of the preceding embodiments, wherein the comparison (e.g., decreasing, increasing, enriching, negative enriching, positive enriching, etc.) is a reference assessment of the collected sample after early administration or delivery.
2395. The method of any of the preceding embodiments, wherein the comparison (e.g., decreasing, increasing, enriching, negative enriching, positive enriching, etc.) is a reference assessment after administration or delivery of the reference agent.
2396. The method of any of the preceding embodiments, wherein the comparison (e.g., decreasing, increasing, enriching, negative enriching, positive enriching, etc.) is a reference assessment of the collected sample after administration or delivery of the reference agent.
2397. The method of any one of embodiments 2395 to 2396, wherein the reference agent is a therapeutic agent.
2398. The method of any one of embodiments 2395 to 2396, wherein the reference agent is an inactive control agent.
2399. The method of any one of embodiments 2395 to 2398, wherein the administering, delivering, and/or assessing is performed under comparable conditions.
2400. A pharmaceutical agent, compound or composition prepared and/or characterized by the method of any one of the preceding embodiments.
2401. The agent, compound or composition of any of the preceding embodiments, prepared and/or characterized by the method of any of the preceding embodiments.
Examples
Those of skill in the art will appreciate that a variety of techniques may be used to prepare and evaluate the provided agents, including a variety of peptides (e.g., stapled peptides) according to the present disclosure, for example, a number of techniques for preparing small molecules and peptides may be utilized to prepare the provided agents, and a variety of assays may be used to evaluate the characteristics and/or activity of the provided agents. Some of these available techniques are described below. As demonstrated herein, in some embodiments, it was determined that the provided techniques can exhibit nanomolar cell-based activity in protein-protein interactions (PPI), transcriptional regulation, proliferation assays, and the like. In some embodiments, it is determined that the provided technology has favorable pharmacokinetic properties. In some embodiments, in vivo administration of the provided technology determines a mid-target (on-target) pharmacodynamic modulation of β -catenin activity and strong anti-tumor activity in various human xenograft models, which determines that the provided technology is useful for treating various conditions, disorders, or diseases as described herein.
Example 1 peptide synthesis.
Peptides can be prepared in particular according to the present disclosure using a variety of peptide synthesis techniques. In many embodiments, the peptides are generally prepared using Fmoc-based synthesis on a suitable solid phase. For a variety of stapled peptides, the amino acid residues are stapled by a suitable chemical reaction (e.g., olefin metathesis of an amino acid comprising an olefin group). Those skilled in the art will appreciate that other suitable techniques may also be used for stapling in accordance with the present disclosure, such as those described in WO/2019/051327, WO/2020/04270, etc., peptide staple-like structures and techniques for preparing peptides are incorporated herein by reference.
For example, in some embodiments, peptides are synthesized on a Liberty Blue peptide synthesizer using standard Liberty Blue conditions on Rink Protide amide resin (primary formamide), ethylindole AM resin (ethylamide), amino alcohol 2-chlorotrityl resin (amino alcohol), or Wang resin with preloaded C-terminal amino acid (carboxylic acid), with 1M DIC in DMF and 1M Oxyma in DMF. All amino acids were coupled singly except that residues after stapling the amino acids and B5 were coupled doubly. The N-terminal residue was subjected to final Fmoc deprotection and endcapped (e.g., acetate endcapped) at room temperature by treating the resin with a suitable endcapping agent (e.g., 5% acetic anhydride, 2.5% diisopropylethylamine, and 92.5% NMP for acetate endcapping) for 30 minutes. The non-acetate amide end caps have a suitable amount of reagent attached, such as 5 equivalents of carboxylic acid, 5 equivalents of DIC, and 5 equivalents of Oxyma in a suitable solvent (e.g., DMF).
The lactam staple structure and the triazole staple structure are blocked prior to olefin metathesis. The lactam staple structure is produced by incorporating an amino-containing residue as an Alloc-protected amino acid and a carboxylate-containing residue as an allyl-protected amino acid. By using 10mol% Pd (Ph) 3 P) 4 The peptide was treated with 10 equivalents of morpholine, phenylsilane or dimethylbarbituric acid for 1 hour at room temperature for Alloc/allyl deprotection. Lactam formation was performed by treating the resin with 10 equivalents of Oxyma and 10 equivalents of DIC at 40℃for 2 hours, then draining and washing the resin with DMF.
During the linear synthesis of peptides, triazole staple-like structures are created by incorporating both azide-containing amino acids and alkyne-containing amino acids. Triazole ring closure was performed by treating acylated linear peptide with copper (II) sulfate (2 equivalents) and sodium ascorbate (2 equivalents) in a t-butanol/water (2/1) mixture. The mixture was heated in the microwave at 80 ℃ for 30 minutes and the resin was then filtered off, followed by washing with DMF and methanol.
Olefin metathesis is carried out by treating the peptide under suitable conditions with a suitable metathesis catalyst, in some embodiments optionally with 30mol% grubb's first generation catalyst in dichloroethane (CAS 172222-30-9) treating multiple cycles (4 cycles) at 40 ℃ for 2 hours, and washing the resin with dichloroethane after each treatment.
The hydrogenation of the peptide staple was carried out by treating the resin with fresh 30mol% Grubb's first generation catalyst (CAS 172222-30-9) in 1, 2-dichlorobenzene. Triethylsilane (50 eq.) was added and the resin was placed in a heated shaker at 50 ℃ overnight and then washed with dichloroethane.
Peptide cleavage was performed by treating the resin with 95% trifluoroacetic acid and 5% triisopropylsilane for 1 hour, and the crude peptide was precipitated in diethyl ether. Purification was performed by preparative HPLC with MS detection and Waters XSelect CSH C18 column using water with 0.1% formic acid and acetonitrile with 0.1% formic acid. In general, if the isomers are identified and separated by HPLC purification, they are separated and tested separately by elution peaks (e.g., UV at 220 nm), otherwise the peptides are separated as a combination (typically based on HPLC peaks) and tested (all peptides within a single HPLC peak are typically tested together in a single composition).
Amino acids suitable for synthesis are commercially available or may be prepared according to the present disclosure. Certain amino acids and their preparation/formulations are described below: the priority applications WO 2022/020651 or WO 2022/020652, for example, the preparation of: (S) -2- ((((9H-fluoren-9-yl) methoxy) carbonyl) amino) -3- (2- (tert-butoxycarbonyl) phenyl) propanoic acid, (S) -3- (2- ((((9H-fluoren-9-yl) methoxy) carbonyl) amino) -3- (benzyloxy) -3-oxypropyl) benzoic acid tert-butyl ester, tfeGA, and the like, amino acids, and their preparation/formulations (including methods, reagents, intermediates, and the like), each of which is independently incorporated herein by reference.
Some peptide formulations are shown below as examples.
Compounds having a staple-like structure bridging the substituted glutamine residues between AA7 and AA14 were synthesized in the following manner: fmoc-BztA-Glu (OAllyl) -protide resin was synthesized on Liberty Blue as described above. By using allyl in DCE 10% Pd (Ph 3 P) 4 And 10 equivalents of phenylsilane were treated at room temperature for 1 hour to effect deprotection. The mono-alloc protected diamine was coupled with the deprotected Glu residue by treating the resin with 4 equivalents of the protected diamine, 4 equivalents of DIC and 4 equivalents of Oxyma in DMF at 40℃for 2 hours. The resin was then washed with DMF and loaded back into Liberty Blue and the linear peptide sequence with Glu (OAllyl) at position 7 was completed. The resin was acetyl-terminated as described above. Alloc/allyl deprotection was performed by treating the peptide with 10mol% Pd (Ph 3P) 4 plus 10 equivalents of morpholine, andthe lactamization was performed by treating the resin with 10 equivalents DIC and 10 equivalents Oxyma in DMF at 40 ℃. Ring closure metathesis, cleavage and purification were performed as described above.
I-45, I-46, I-47, I-48, I-49, I-50, I-51, I-52, I-53, I-54: compounds having a staple-like structure between lysine residues at positions 7 and 14 are produced in the following manner: the linear sequence was synthesized on Liberty Blue as described above, incorporating Fmoc-Lys (ivDde) -OH at positions 7 and 14. Following acetyl capping and olefin metathesis, the ivDde groups were removed by treatment with 5% hydrazine in DMF at 40 ℃ for two cycles (30 min), followed by washing with DMF. The resin was then treated with 2 equivalents of diacid in DMF, 5 equivalents of DIC and 5 equivalents of Oxyma at 40℃for 2 hours. The resin was then washed with DMF and DCE, and cleaved and purified as described above.
I-303, I-517, I-518: biotinylated peptides were generated by incorporating Fmoc-Lys (ivDde) -OH in the linear sequence. Following acetyl capping and olefin metathesis, the ivDde groups were removed by treatment with 5% hydrazine in DMF at 40 ℃ for two cycles (30 min), followed by washing with DMF. The resin was then treated with 3 equivalents of biotin-PEG 8-acid (CAS 2143964-62-7), 3 equivalents of HATU, 10 equivalents of diisopropylethylamine in DMF for 2 hours at 40 ℃. The resin was then washed with DMF and DCE, and cleaved and purified as described above.
I-606, I-607: peptides with azido lysines in the final sequence were generated by incorporating Fmoc-Lys (ivDde) -OH in the linear sequence. After olefin metathesis, the ivDde group was removed by treatment with 5% hydrazine in DMF at 40 ℃ for two cycles (30 min) followed by washing with DMF. The resin was then treated with 3 equivalents of 1H-imidazole-1-sulfonyl azide sulfate in DMF (CAS 1357503-23-1), 9 equivalents of diisopropylethylamine and 0.5 equivalents of copper (II) sulfate pentahydrate at 40℃for 3 hours. The resin was then washed with DMF, water, DMF and DCE, and cleaved and purified as described above.
The cysteine-containing staple structure is blocked after olefin metathesis, peptide cleavage and purification. In a vial, the purified biscysteine peptide was dissolved in DMF and 5 equivalents of dibromo linker were added followed by 100mM ammonium bicarbonate pH 8 buffer followed by DTT (10 mM). After stapling was completed, the crude reaction mixture was purified by preparative HPLC as described above.
I-469: synthesis on Rink amide resin
Ac-PL3-OAsp-Npg-B5-Asp-3COOHF-Aib-Ala-Phe-Lys 3-PyrS2-2F3MeF-BztA-GlnR 3-Ala-protide resin and a lactam staple structure was mounted as above. A plastic syringe containing 200mg of the resin-bound peptide containing the free N-terminal amine was swollen in 0.5mL DMF. To the swollen resin was added a solution of (2S) -4- (tert-butoxy) -2-hydroxy-4-oxobutanoic acid (85.5 mg,0.45 mmol) in 0.5mL DMF, 450. Mu.L 1M DIC and 450. Mu.L 1M Oxyma. The syringe was shaken for 90 minutes at room temperature. The resin was then washed with DMF, DCM, meOH and then with DCM and subsequently dried under vacuum. The resin bound peptide was swollen in 0.5mL DCM. To the swollen resin was added 500. Mu.L of 0.1M DMAP in DCM followed by a solution of PL3-Ac (88.75 mg 0.45 mmol) and DCC (92.8 mg 0.45 mmol) in DCM. The syringe was shaken at 40℃for 3 hours. The resin was then washed with DMF, DCM, meOH and then with DCM and subsequently dried under vacuum. Ring closure metathesis, peptide cleavage and purification were then performed as described above.
I-427: synthesis on Rink amide resin
Fmoc-R5-Asp-3COOHF-Aib-Ala-Phe-Lys 3-pyrS2-3Thi-BztA-GlnR 3-Ala-protide resin and mounting a lactam staple structure as described above. On a polypropylene syringe equipped with a porous polypropylene disk at the bottom, 0.05mmol (about 0.18 g) of intermediate 1 on resin was swollen on DCE for 15 minutes. The ring closure metathesis between R5 and the side chain of pyrS2 (ring closing metathesis, RCM) was carried out under standard protocols (30 mol% Grubbs I catalyst, 2X 2 hours at 40 ℃, in DCE). The resin was then washed with DCE 2×, DMF 2×, DCM, meOH and DCM. Next, the resin was swollen in DMF for 15 min, treated with 20% piperidine in DMF 2 x for 25 min and washed with DMF 5 x and NMP 2 x. To the swollen resin was added a pre-activated mixture of Fmoc-AllylGly-OH (5 eq), oxyma (5 eq) and DIC (5 eq) in NMP (0.4M). The mixture was shaken at room temperature for 2 to 3 hours. The chloranil test indicated complete coupling. The resin was then washed with DMF 5X and the above cycle was repeated with Fmoc-Asp (OtBu) -OH, fmoc- αMePro-OH and 4-pentenoic acid. The second RCM between the N-terminal end-capping of 4-pentenoic acid and the side chain of AllylGly was now carried out under standard protocol (30 mol% Grubbs I catalyst, 40 ℃, 2X 2 hours in DCE). The resin was then washed with DMF 4×, DCM 3×, meOH, DCM and dried under high vacuum.
The peptide was cleaved from the resin with 3ml TFA/H2O/TIPS (95:2.5:2.5) at room temperature for 2 hours, then precipitated by cold diethyl ether, and the obtained residue was applied to a reverse phase HPLC column to give the title compound (1.5 mg) as a white powder after lyophilization of the pure fractions.
I-429: the same experimental procedure as described for I-427 was used in this synthesis. The only difference is the coupling of 5-hexenoic acid at the last step of the linear peptide synthesis. The peptide was cleaved from the resin with 3ml TFA/H2O/TIPS (95:2.5:2.5) at room temperature for 2 hours, then precipitated by cold diethyl ether, and the obtained residue was applied to a reverse phase HPLC column to give the title compound (1.3 mg) as a white powder after lyophilization of the pure fractions.
I-428: starting from Fmoc-R5-Asp-3COOHF-Aib-Ala-Phe-Lys 3-pyrS2-3Thi-BztA-GlnR 3-Ala-protide resin, 0.05mmol (about 0.18 g) of intermediate 1 on resin was swollen on DCE for 15 minutes in a polypropylene syringe equipped with a porous polypropylene disk at the bottom. The Ring Closure Metathesis (RCM) between R5 and the side chain of pyrS2 was carried out by standard protocols (30 mol% Grubbs I catalyst, 2X 2 hours at 40℃in DCE). After the resin was washed with DCE 2×, DMF 2×, DCM, meOH and DCM, it was swollen in 1, 2-Dichlorobenzene (DCB) for 15 min. The solvent was drained and about 15mg of Grubbs I catalyst as a solid was added to the resin. The syringe was closed with a moving plunger (pluringer) and triethylsilane (50 eq) and DCB (0.6 mL) were then added to the mixture through the needle. The syringe was shaken at 50 ℃ for 18 hours to produce the corresponding i+7 reduced staple-like structure. The resin was then washed with DMF 4×, DCM 3×, meOH, DCM and DMF. Next, the resin was swollen in DMF for 15 min, treated with 20% piperidine in DMF 2 x for 25 min and washed with DMF 5 x and NMP 2 x. To the swollen resin was added a pre-activated mixture of Fmoc-AllylGly-OH (5 eq), oxyma (5 eq) and DIC (5 eq) in NMP (0.4M). The mixture was shaken at room temperature for 2 to 3 hours. The chloranil test indicated complete coupling. The resin was then washed with DMF 5X and the above cycle was repeated with Fmoc-Asp (OtBu) -OH, fmoc- αMePro-OH and 4-pentenoic acid. The second RCM between the N-terminal end-capping of 4-pentenoic acid and the side chain of AllylGly was now carried out by standard protocol (30 mol% Grubbs I catalyst, 40 ℃, 2X 2 hours in DCE). The resin was then washed with DMF 4×, DCM 3×, meOH, DCM and dried under high vacuum. The peptide was cleaved from the resin with 3ml TFA/H2O/TIPS (95:2.5:2.5) at room temperature for 2 hours, then precipitated by cold diethyl ether, and the obtained residue was applied to a reverse phase HPLC column to give the title compound (5.2 mg) as a white powder after lyophilization of the pure fractions.
I-431: the same experimental procedure as described for I-428 was used in this synthesis. The only difference is the coupling of 5-hexenoic acid at the last step of the linear peptide synthesis. The peptide was cleaved from the resin with 3ml TFA/H2O/TIPS (95:2.5:2.5) at room temperature for 2 hours, then precipitated by cold diethyl ether, and the obtained residue was applied to a reverse phase HPLC column to give the title compound (0.85 mg) as a white powder after lyophilization of the pure fractions.
I-425: starting from Fmoc-R5-Asp-3COOHF-Aib-Ala-Phe-Lys 3-PyrS2-3Thi-BztA-GlnR 3-Ala-protide resin, 0.05mmol (about 0.18 g) of the resin upper intermediate precursor, in which no staple structure (e.g., (i, i+7) between R5 and PyrS 2) had formed, was swollen in DMF for 15 minutes, treated with 20% piperidine in DMF 2 x, and washed with DMF 5 x and NMP 2 x in a polypropylene syringe equipped with a porous polypropylene disk at the bottom. To the swollen resin was added a pre-activated mixture of Fmoc-AllylGly-OH (5 eq), oxyma (5 eq) and DIC (5 eq) in NMP (0.4M). The mixture was shaken at room temperature for 2 to 3 hours. The chloranil test indicated complete coupling. The resin was then washed with DMF 6X and the above cycle was repeated with Fmoc-Asp (OtBu) -OH, fmoc- αMePro-OH and 4-pentenoic acid. The simultaneous RCM between the N-terminal end-capping of 4-pentenoic acid and the side chain of AllilGly and between the side chain of amino acid R5 and pyrS2 was carried out by standard protocols (30 mol% Grubbs I catalyst, 40 ℃, 4X 2 hours in DCE). Thereafter, the resin was washed thoroughly with DCE, DCM, DMF, meOH and DCM and dried under high vacuum for 3 to 4 hours. The resin was swollen in 1, 2-Dichlorobenzene (DCB) for 15 minutes. The solvent was drained and about 15mg of Grubbs I catalyst as a solid was added to the resin. The syringe was closed with a moving plunger, and triethylsilane (50 eq) and DCB (0.6 mL) were then added to the mixture through the needle. The syringe was shaken at 50 ℃ for 18 hours to yield the corresponding fully reduced analog. The resin was then washed with DMF 4×, DCM 3×, meOH, DCM and dried under high vacuum. The peptide was cleaved from the resin with 3ml TFA/H2O/TIPS (95:2.5:2.5) at room temperature for 2 hours, then precipitated by cold diethyl ether, and the obtained residue was applied to a reverse phase HPLC column to give the title compound (2 mg) as a white powder after lyophilization of the pure fractions.
I-426: the same experimental procedure as described for I-425 was used in this synthesis. The only difference is the coupling of 5-hexenoic acid in the last step of the linear peptide synthesis. The peptide was cleaved from the resin with 3ml TFA/H2O/TIPS (95:2.5:2.5) at room temperature for 2 hours, then precipitated by cold diethyl ether, and the obtained residue was applied to a reverse phase HPLC column to give the title compound (1.5 mg) as a white powder after lyophilization of the pure fractions.
I-471: starting from Fmoc-R5-Asp-3COOHF-Aib-Ala-Phe-Lys 3-PyrS2-3Thi-BztA-GlnR 3-Ala-protide resin, 0.05mmol (about 0.18 g) of the resin upper intermediate precursor, in which no staple structure (e.g., (i, i+7) between R5 and PyrS 2) had formed, was swollen in DMF for 15 minutes, treated with 20% piperidine in DMF 2 x, and washed with DMF 5 x and NMP 2 x in a polypropylene syringe equipped with a porous polypropylene disk at the bottom. To the swollen resin was added a pre-activated mixture of Fmoc-AllylGly-OH (5 eq), oxyma (5 eq) and DIC (5 eq) in NMP (0.4M). The mixture was shaken at room temperature for 2 to 3 hours. The chloranil test indicated complete coupling. The resin was then washed with DMF 6 x. The above cycle was repeated with Fmoc-Asp (OtBu) -OH, fmoc- αMePro-OH and 2- (prop-2-en-1-yloxy) -benzoic acid. Simultaneous Ring Closure Metathesis (RCM) between the N-terminal end cap of benzoyl-O-allyl and the side chain of AllylGly and between amino acid R5 and the side chain of pyrS2 was carried out by standard protocols (30 mol% Grubbs I catalyst, 40 ℃,3x 3 hours in DCE). The resin was washed thoroughly with DCE, DCM, DMF, meOH and DCM and dried under high vacuum for 3 to 4 hours. The peptide was cleaved from the resin with 3ml TFA/H2O/TIPS (95:2.5:2.5) at room temperature for 2 hours, then precipitated by cold diethyl ether, and the obtained residue was applied to a reverse phase HPLC column to give the title compound (0.83 mg) as a white powder after lyophilization of the pure fractions.
I-519: starting from Fmoc-R5-Asp-3COOHF-Aib-Ala-Phe-Lys 3-PyrS2-3Thi-BztA-GlnR 3-Ala-protide resin, 0.05mmol (about 0.18 g) of the resin upper intermediate precursor, in which no staple like structure (e.g., (i, i+7) between R5 and PyrS 2) had been formed, was swollen in DCE for 15 minutes in a polypropylene syringe equipped with a porous polypropylene disc at the bottom. The ring-closing metathesis (RCM) between R5 and the side chain of Pyrs2 was carried out by standard protocols (30 mol% Grubbs I catalyst, 40 ℃,2X 2 hours in DCE). The resin was then washed with dce3×and DMF 3×. Thereafter, the resin was swelled in DMF for 15 min, treated with 20% piperidine in DMF for 2X 25 min and washed with DMF 5X and NMP 2X. To the swollen resin was added a pre-activated mixture of Fmoc-Dap (ivDde) -OH (5 eq), oxyma (5 eq) and DIC (5 eq) in NMP (0.4M). The mixture was shaken at room temperature for 2 to 3 hours. The resin was then washed with DMF 6X and the above cycle was repeated with Fmoc-Asp (OtBu) -OH, fmoc- αMePro-OH and 5-hexenoic acid. The ivDde protecting group on the diaminopropionic acid (Dap) side chain was removed by treating the DMF swollen resin with a 5% solution of hydrazine in DMF at 40 ℃ for 2x 20 min. After that, the resin was washed thoroughly with DMF and 2 XNMP. O-nitrobenzenesulfonyl chloride (4 eq) and 2,4, 6-collidine (4 eq) in NMP were added to the swollen resin and the reaction was shaken at room temperature for 30 minutes to give the desired N-activated intermediate. The resin was washed thoroughly with DMF. N-alkylation of activated primary amine was performed with allyl bromide (15 eq) and DBU (15 eq) in DMF and the resin was shaken at room temperature for 2 days. The resin was washed thoroughly with DMF and 2 XNMP. To drive the N-alkylation reaction to completion, the resin was treated with allyl bromide (15 eq.) and 2, 6-lutidine (15 eq.) in NMP under microwave conditions at 110℃for 30 minutes. The resin was washed thoroughly with DMF and 2×nmp and cleavage by LCMS and analysis showed complete reaction. The N-activating group (oNBS) was removed by treating the resin (2X 20 min at room temperature) with mercaptoethanol (10 eq.) and DBU (5 eq.) in NMP. The secondary amine obtained at the Dap side chain was alkylated with benzyl bromide (10 eq.) and 2, 6-lutidine (15 eq.) at 110℃for 2X 25 min under microwave conditions. The second RCM between the N-terminal end-capping of 5-hexenoic acid and the side chain of Dap (allyl) was performed by standard protocol (30 mol% Grubbs I catalyst, 40 ℃,2X 2 hours in DCE). The resin was washed thoroughly with DCE, DCM, DMF, meOH and DCM and dried under high vacuum for 3 to 4 hours. The peptide was cleaved from the resin with 3ml TFA/H2O/TIPS (95:2.5:2.5) at room temperature for 2 hours, then precipitated by cold diethyl ether, and the obtained residue was applied to a reverse phase HPLC column to give the title compound (0.55 mg) as a white powder after lyophilization of the pure fractions.
I-520: the same experimental procedure as described for I-519 was used in this synthesis. The only difference was that the secondary amine generated at the Dap side chain was acylated (instead of alkylated) at room temperature for 2 to 3 hours using benzoic acid (5 equivalents), oxyma (5 equivalents) and DIC (5 equivalents) in NMP. The peptide was cleaved from the resin with 3ml TFA/H2O/TIPS (95:2.5:2.5) for 2 hours at room temperature, then precipitated by cold diethyl ether, and the obtained residue was applied to a reverse phase HPLC column to give the title compound (0.62 mg) as a white powder after lyophilization of the pure fractions.
I-564: the same experimental procedure as described in I-519 was used in this synthesis, with two important changes. The first is to acylate (instead of alkylating) the secondary amine generated at the Dap side chain with pivaloyl chloride (7 eq.) and NMM (10 eq.) under microwave conditions at 77℃for 15 minutes. Second, the resin was swollen in 1, 2-Dichlorobenzene (DCB) for 15 minutes, followed by the addition of about 15mg Grubbs I catalyst as a solid after the solvent was drained. The syringe was closed with a moving plunger and triethylsilane (50 eq.) and about 0.6mL of DCB were added to the mixture through the needle. The syringe was shaken at 50 ℃ for 18 hours to yield the corresponding fully reduced analog. The resin was then washed with DMF 4×, DCM 3×, meOH and DCM and dried under high vacuum. The peptide was cleaved from the resin with 3ml TFA/H2O/TIPS (95:2.5:2.5) at room temperature for 2 hours, then precipitated by cold diethyl ether, and the obtained residue was applied to a reverse phase HPLC column to give the title compound (1.36 mg) as a white powder after lyophilization of the pure fractions.
I-565: the same experimental procedure as described for I-564 was used in this synthesis. The only difference was that the secondary amine generated at the Dap side chain was acylated for 2 to 3 hours at room temperature using cyclohexane carboxylic acid (5 equivalents), oxyma (5 equivalents) and DIC (5 equivalents) in NMP. The peptide was cleaved from the resin with 3ml TFA/H2O/TIPS (95:2.5:2.5) at room temperature for 2 hours, then precipitated by cold diethyl ether, and the obtained residue was applied to a reverse phase HPLC column to give the title compound (1.19 mg) as a white powder after lyophilization of the pure fractions.
I-562: an experimental procedure similar to that described in I-519 was used in this synthesis, but with three important variations. First, 4-pentenoic acid was used as the N-terminal end capping group. Second, the secondary amine at the Dap side chain was N-alkylated with benzyl bromide (10 eq.) and 2, 6-lutidine (15 eq.) at 110℃for 2X 25 minutes under microwave conditions. Third, both olefin staple-like structures are reduced simultaneously. The resin was swollen in 1, 2-Dichlorobenzene (DCB) for 15 minutes, the solvent was drained, and about 15mg of Grubbs I catalyst as a solid was added to the resin. The syringe was closed with a moving plunger and triethylsilane (50 eq) and DCB (0.6 mL) were added to the mixture through the needle. The syringe was shaken at 50 ℃ for 18 hours to yield the corresponding fully reduced analog. The resin was then washed with DMF 4×, DCM 3×, meOH and DCM and dried under high vacuum. The peptide was cleaved from the resin with 3ml TFA/H2O/TIPS (95:2.5:2.5) at room temperature for 2 hours, then precipitated by cold diethyl ether, and the obtained residue was applied to a reverse phase HPLC column to give the title compound (0.44 mg) as a white powder after lyophilization of the pure fractions.
I-563: the same experimental procedure as described for I-562 was used in this synthesis. The only difference was that the secondary amine generated at the Dap side chain was acylated (instead of alkylated) at room temperature for 2 to 3 hours using benzoic acid (5 equivalents), oxyma (5 equivalents) and DIC (5 equivalents) in NMP. The peptide was cleaved from the resin with 3ml TFA/H2O/TIPS (95:2.5:2.5) at room temperature for 2 hours, then precipitated by cold diethyl ether, and the obtained residue was applied to a reverse phase HPLC column to give the title compound (1.19 mg) as a white powder after lyophilization of the pure fractions.
Mass spectrometry was performed as follows: 2uL of 200uM peptide solution in DMSO was injected over a gradient of 95/5 water/acetonitrile to 5/95 water/acetonitrile over a Waters Acquity UPLC-MS system with a 2.1×50mm, 1.7 μm CSH C18 column at 40 ℃ at a flow = 0.6 mL/min. Product peaks were analyzed in both positive and negative ionization modes.
Example 2 the techniques provided may provide improved properties and/or activity.
In some embodiments, solubility is assessed. In some embodiments, the following illustrates a useful scheme as an example: 50uM peptides were incubated in 99.5% PBS/0.5% DMSO at 37℃for 15 min. After ultracentrifugation of the PBS solution, the supernatant was analyzed by HPLC and compared to HPLC injection of 50uM peptide DMSO solution. The solubility is determined by: [ (PBS peak area)/(DMSO peak area) ]. In some embodiments, the provided agent (e.g., stapled peptide) has a solubility of about or at least about 1 to 50, 10, 20, 30, 40, or 50uM as measured using such a protocol.
In some embodiments, the log d of the provided agent (e.g., stapled peptide) is assessed. In some embodiments, shake flask LogD is evaluated using the following procedure as an example. In some embodiments, the shake flask LogD of certain medicaments (e.g., stapled peptides) is about 0 to 3, 0.1 to 2.5, 0.5 to 2, 1 to 2, 1.5 to 2, or about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, or 3.
The required instrument:
EP pipettor (EP Motion)
4titude board sealer
Sample scanner
Hamilton cap remover
Eppendorf centrifuge
Eppendorf vibrator
Acoustic wave instrument
Agilent single quadrupole HPLC-MS
The required materials are as follows:
eppendorf 384 well 100uL volume plate
Eppendorf 96 well 1mL plate
Eppendorf 96 well 500uL plate
EP pipettor
EP pipettes 50uL, 300uL, 1000uL tips (tips).
Preparation for plate generation:
1. a tray aliquot was removed from compound management.
2. Up to 45 compounds were used and the last 3 spots were left as aliquots of the standard.
3. An aliquot plate on a sample scan (SampleScan) was scanned.
4. And obtaining a scanning value and generating an excel file.
5. Generating a csv file from the exported excel
a. The csv should comprise two columns
i. The first column: sample position
Second column: sample name.
6. The compound was spun in a centrifuge at 3000rpm for 15 seconds.
7. The peer-to-peer aliquots were uncapped using a Hamilton uncapping machine.
8. An aliquot in the tray was placed into an EP pipette.
Plate generation:
1. selection on EP pipettes:
a.Home＞Chemistry＞logDPart1v1_100mL_NoSTD。
2. aliquots, tips, plates (96 well 1mL Eppendorf plates) and reservoirs were placed according to the instrument.
a. The reservoir contained presaturated octanol at pH 7.4 and presaturated buffer at pH 7.4.
3. Ensure that the total number of samples read out to 48.
4. Select run and ensure that "test volume" is selected, you can deselect "check tip" and "laboratory instrument placement".
5. The operation method.
6. The completed 96-well plate was removed. The EP pipettor was cleaned.
7. The completed plate was placed into compound management and the 4titude plate sealer was opened.
a. Wait until the plate sealer shows a temperature of 170C.
b. The silver flake is placed on a plate and held in place with a gold holder.
c. A selection operation, placing the plate in the holder, and the holder in place. The operation is pressed again.
d. Once the plate is ejected, it is firmly sealed using rollers.
8. The plate was turned over and placed on an Eppendorf shaker at 2000rpm for 1 hour.
9. The plate was removed and sonicated for 10 minutes.
10. Centrifuge at 3000rpm for 10 minutes.
Plate generation: final result
1. Selection on EP pipettes:
a.Home＞Chemistry＞logDPart2v1_80mL
2. aliquots, tips, plates (96-well 1mL Eppendorf plates, 96-well 500mL Eppendorf plates, 384-well 100 μl plates (final)) and reservoirs were placed according to the instrument.
a. The reservoir contained (50/50) pre-saturated octanol/DMSO at pH 7.4, DMSO, acetonitrile and pre-saturated buffer at pH 7.4.
3. Ensure that the total number of samples read out to 48.
4. Select run and ensure that "test volume" is selected, you can deselect "check tip" and "laboratory instrument placement".
5. The operation method.
6. The completed 96-well plates and the final 384-well plates were removed. The EP pipettor was cleaned.
7. Two 96-well plates were sealed with a rubber plate sealer and stored in a 4C freezer.
8. The finished final plate was placed into compound management and the 4titude plate sealer was opened.
a. Wait until the plate sealer shows a temperature of 170C.
b. The pierceable silver flake is placed on the plate and the gold holder is used to secure the silver flake in place.
c. A selection operation, placing the plate in the holder, and the holder in place. The operation is pressed again.
d. Once the plate is ejected, it is sealed firmly using a roller (roll).
9. Go to Agilent HPLCMS.
Plate programming:
1. the chemical workstation (Chemstation) on Agilent HPLCMS was opened.
2. Buffer C (water with 0.1% formic acid), D (acetonitrile with 0.1% formic acid) and wash solution (MeOH) were ensured to be full.
3. The green on button is clicked to allow the instrument sufficient time to equilibrate while the sequence is being programmed.
4. Clicking the sequence button in the top drop down menu:
a. selection of a New sequence
b. The sequence was saved as yyymmdd_sol.
5. From the list of sequences:
a. the sample is selected for introduction.
b. Click browse and find the previously created csv file.
c. Clicking on the next step and then clicking is completed.
6. Opening sequence:
a. the columns that only need to be filled are the positions:
b. and sample name:
7. double-click method box:
a. selection of 10_mincd 60-95
8. For injection volume input 10, 10uL will be injected.
9. 2 is entered in the number of injections, representing 2 injections/well.
10. The column containing the method, injection volume and number of injections is now highlighted and dragged down to the bottom of the sequence. Pressing Ctrl and right clicking selects the down fill.
11. Blank samples are added in the first and last slots (slots).
a. D1B-D1 is entered for the sample location.
b. Blanks are entered for sample names.
c. The same method 10_mincd 60-95 is entered.
d. Input 10 is taken as the injection volume.
e. Input 4 is the number of injections.
12. Right click on a widget (widget) of the mass spectrometer and enter the mass range of the compound selected for operation.
13. Clicking starts.
And (3) data processing:
1. an offline version of the chemical workstation software is opened and the desired sequence is selected by date.
2. Double-clicking on the row containing the compound of interest. This will give a chromatogram.
a. A delimiting tool is selected to remove any automatic integration (automatic integration).
b. The mass average chromatographic tool is selected and dragged onto the peak of interest.
c. The expected quality is found.
d. The peak bounding tool is returned again and dragged over the peak containing the quality of interest to integrate the peak.
e. The same procedure was performed for the second preparation of the same compound.
3. The integrated compound results were derived as pdf.
4. The integrated area is obtained and entered into an entry table in excel.
5. Once all results are entered, the function in excel will automatically calculate the average log and standard deviation.
a. To illustrate the dilution during the entire plate creation process, the final calculation of Logd is as follows:
log = log ((octanol peak x 40)/(buffer peak x 2)).
The average log is the average of the log values calculated for peaks 1 and 2 and is also the standard deviation of the log values calculated for peaks 1 and 2.
Special cases: logd seen only in octanol phase is indicated as > 0, logd seen only in buffer phase is indicated as < 0.
if no compound is visible in either phase, it may indicate that there is a solubility problem. Generally labeled Div/0-! And the observation is contained in the comment section.
Some results are shown herein as examples.
Example 3. A variety of provided peptides can bind to β -catenin.
As will be appreciated by those of skill in the art, many techniques may be utilized in accordance with the present disclosure to assess binding to a target (e.g., β -catenin). Some useful techniques and results are described below as examples.
In some embodiments, the assay is fluorescence polarization. Usable schemes are described below as examples.
Fluorescence polarization IC50: peptide solutions were serially diluted 3-fold in 90% DMSO using moquito (SPT) and 40nL of titrated peptide was added to 20uL buffer (50mM HEPES,pH 7.5, 125mM NaCl,2% glycerol, 0.5mM EDTA,0.05%v/v pluronic acid) to a final concentration of 10uM to 5nM by multitrop TM Combi (Thermo Scientific) it was plated into black polystyrene 384 well plates (Corning). Preparation of a Probe solution (in buffer with 10nM of 5 FAM-labeled TCF4 No. 10 to10nM full-length beta-catenin (Uniprot ID P35222) mixed with 53-residue (Uniprot ID Q9NQB 0) peptide and using Multidrop TM Combi (Thermo Scientific) plating was performed at 20uL per well. Plates were incubated at 20℃for 60 min in the absence of light and subsequently read. Reads were performed in duplicate on a CLARIOstar reader (BMG Labtech) and data were fitted to a 1:1 binding model with hill slope using internal script. All provided concentrations are final concentrations. Some of the results are shown as examples in table E1 below.
In some embodiments, binding to β -catenin may be measured by surface plasmon resonance (surface plasmon resonance, SPR). Usable schemes are described below as examples. Various agents (such as those shown as examples in E2) exhibit binding to β -catenin, which in some embodiments has a low or sub-nM Kd; other values (e.g. t 1/2 ) It can also be evaluated and in many cases evaluated.
Dilution of peptides at 10mM concentration in DMSO to Biacore TM Buffer (50mM Tris pH 8.0, 300mM NaCl,2% glycerol, 0.5mM TCEP,0.5mM EDTA,0.005% tween-20,0.09% DMSO) was run to give the appropriate dilution range. Then use Biacore TM Biotin CAPTURE kit (GE Healthcare) in Biacore TM These diluted peptide samples were analyzed on S200, the kit having been functionalized with biotinylated β -catenin at residues 134 to 665 (Uniprot ID P35222). Biacore fitted to 1:1 binding model was used TM The results were analyzed by the weight evaluation software.
Example 4. The techniques provided can modulate interactions with beta-catenin in cells.
The characteristics and/or activity of a provided compound (e.g., a stapled peptide) can be evaluated in a cell using a variety of techniques. In some embodiments, a useful assay is a Nano-BRET target binding assay that evaluates β -catenin/TCF 4 binding. Usable schemes are described below as examples.
On day 1 HEK293 cells were seeded. Cells were used at about 70% confluence. Trypsin elimination without PBS washCells are lysed (e.g., 5ml trypsin/75 flasks, for 2 to 5 minutes at room temperature). Trypsin was quenched with 10 mM MEM medium. Cells were transferred to falcon tubes. Spin down (spin down) at 250g for 5 minutes at room temperature. The supernatant was discarded. Cells were gently resuspended in 10mLMEM medium. Cells were counted twice and how many cells were needed were calculated. The parental HEK293 cell line was plated using MEM medium in 7M cells/12 ml/75cm2 flasks. The plate was shaken several times to disperse the cells evenly. At 37℃with 5% CO 2 Incubate for 5 hours. Cells should spread evenly and meet approximately 70% after, for example, 5 hours.
Transfection of Nano-BRET construct (B-cat-Halo & TCF 4-Luc): fugen-HD transfection reagent was brought to room temperature. Mixing was done by inversion tube, if the precipitate was visible, then warmed to 37 ℃ and allowed to cool to room temperature. Cell confluence (70% to 80%) in the flasks was examined under a microscope. LiCl (LiCl 30mM working concentration, liCl can be GSK3 inhibitor and reduce β -catenin degradation) was added to the vials containing the cells. Transfection mixtures were prepared in tubes containing assay medium based on manufacturer's instructions (see examples of table below):
transfection mixture preparation
FuGene was added last and mixed gently. No swirling was performed. The transfection mixture was incubated at room temperature for 10 to 15 minutes. If more than one target pair is to be tested, the amount of transfection mixture is calculated using the table above for the other construct pairs. 700uL of transfection mixture was gently added per vial and the plate gently shaken several times. Cells were incubated at 37℃for 18 to 24 hours at 5% CO 2.
On day 2, transfected cells were harvested and re-plated in 384-well plates and pre-distributed medium and compounds in the wells. mu.L of assay medium containing 30mM LiCl was dispensed in all wells of 384 well plates. In some embodiments, a liquid handling system is used to prepare compound plates with a maximum concentration of 10mM and serially diluted 1:3 to a minimum concentration of 13uM. 80nL of these compound series were distributed into 20uL of medium pre-distributed in plates. This produced a 2 x concentration in the wells, which concentration was further diluted once cells were added.
Culture medium from transfected cell flasks was collected in Falcon tubes while compound dilution and partitioning was performed. This is to harvest the floats, as they may still be viable and transfected. Cells were digested with trypsin without washing with PBS (5 ml trypsin/bottle). Trypsin was quenched with 5mL MEM medium. Cells were collected and added to falcon tubes. The flask was washed with 5 to 10mL MEM medium and added to a falcon tube. Rotate at 250g for 5 minutes at room temperature. The supernatant was discarded. The cells were gently resuspended in 5mL of assay medium (optionally containing LiCl). Cells were counted twice and the average count was calculated. Dilution in cell dilution at 1:500NanoBRET TM 618 ligand. In addition to a column of 384 well plates, 20 μl of cell suspension (5,000 cells/40 μl/well) was dispensed per well (plates were used, e.g. Corning Solid White Flat Bottom TC treatment plates). For the last column, 20. Mu.L of cells containing an equal amount of DMSO was added. LiCl concentration was 30mM. The cells were distributed to 20. Mu.L of compound-containing medium to achieve the final working dilution desired. Incubate overnight at 37℃at 5% CO 2.
On day 3, fluorescence was read with a Nano-BRET substrate. The plates were removed from the incubator to room temperature (30 minutes). CTG reagent was also equilibrated to room temperature. The Nano-BRET substrate is diluted 1:100 in assay medium. 10 μl of diluted substrate was added to each well and shaken for 30 seconds. Immediately (within 10 minutes) read on ClarioSTAR or GloMAX. Donor emission at 460 nm. Acceptor emission at 618 nm. The same plate was used to measure Cell viability (Cell Titer-Glo-2.0 (Cell Titer-Glo, CTG) viability test). After reading the BRET signal, CTG reagent was added to each well at a ratio of 1:2 and shaken on an orbital shaker for 2 minutes. Incubate at room temperature for 10 to 30 minutes. Luminescence was read on ClarioSTAR or GloMAX. Analysis was performed using a two-parameter Hill function, and high control (ligand-bearing cells) and low control (ligand-free cells) at R, log (inhibitor) versus response using nonlinear regression to measure absolute IC50 (absic50=x50 ]) for each compound.
Some of the results are shown in table E1 as examples.
Reporter IC50: the activity of the provided technology was also determined in the TCF reporter assay described below. Those skilled in the art will appreciate that other suitable reagents may be used and that various parameters may be adjusted.
On day 1, cells cultured in flasks (e.g., DLD 1) that were no more than about 60% to 70% confluent were washed with PBS and trypsinized in 3ml/T75 until the cells were free floating. Cells were spun at 1100RPM for 5 minutes. After rotation, the supernatant was gently aspirated and the cells were resuspended in 10mL assay medium (4% FBS RPMI or 20% FBS RPMI, depending on the desired serum concentration). Cells were counted twice using a Countess cell counter, the counts were averaged, and cell concentrations were adjusted. The desired seeding density was 2500 cells/well in 40uL of assay medium. Cells were plated in columns 1 to 22 of 384 well white solid bottom plates using a multittop Combi. Cell-free assay medium was added to columns 23 and 24. Assay plates were incubated overnight at 37℃with 5% CO2 at the top rack (back side) of the incubator.
On the next day, the compound was added. Stock solution was 10mM. The liquid handling system is used to prepare a compound dilution and dispense the compound into an assay plate. Compounds were serially diluted 1/2 or 1/3 (depending on the desired assay conditions) in 90% DMSO to generate 7-point dose curves. From the compound plate, 80nL of compound was dispensed directly into the wells of the assay plate to generate a dose curve starting at 20uM and ending at 313nM (1/2 dilution) or 27nM (1/3 dilution). Untreated control wells received only 90% DMSO. Assay plates were incubated overnight at 37℃with 5% CO2 at the top rack (back side) of the incubator.
On day 3, viability was read using cell-titer Fluor (CTF, promega) and TCF activity was read using BrightGlo (Promega). CTF was mixed to 5 x concentration using 35uL substrate with 14mL buffer. Warmed CTF was added directly to columns 1 to 23 of uncooled assay plates using a multittop Combi, 10 uL/well. The assay plates were incubated at 37℃for 2 hours at 5% CO2 on the top plate (back side) of the incubator and then removed. The time interval for removal of the assay plates from the incubator was 5 minutes. The plates were cooled for 40 min, protected from light, and read using the GloMax CTF program (high sensitivity).
After reading CTF, room temperature bright glo was added to columns 1 to 23 of the room temperature assay plate using a multittop Combi, 35 uL/well. Plates were incubated at room temperature for 2 min, protected from light. The plates were then read using a ClarioStar endpoint luminescence reader.
The Log (inhibitor) vs. response (Hill function with two parameters), as well as high control (DMSO-treated cells) and low control (cell-free wells) were analyzed using non-linear regression in R to measure the absolute IC50 (AbsIC 50 = X [50 ]) for each compound.
For a variety of agents (e.g., certain stapled peptides in table E2 or table E3), low or sub-uM IC50 was observed. Some of the results are shown in table E1 as examples.
COLO320DM proliferation assay IC50: in some embodiments, the inhibition of cell proliferation by the provided technology is assessed using a cell line associated with or from certain conditions, disorders, or diseases. In some embodiments, cell proliferation is assessed in COLO320DM cells. In some embodiments, the following procedure is used for evaluation: on day 1, COLO320DM cells cultured in T75 flasks were trypsinized in 3mL 0.25% trypsin/EDTA for 5 min and quenched with 10mLRPMI-1640+4% HI FBS assay medium. Cells were spun at 1200rpm for 5 minutes, cell pellet was collected and resuspended in assay medium at 5000 cells/mL. Cells were dispensed (50 ul,250 cells/well) into three 384 well plates using a Combi liquid handler. Plates were incubated at 37℃for 18 to 22 hours at 5% CO 2. On day 2, the compound was added. A liquid handling system was used to prepare a compound dilution and dispense the compound into an assay plate. Compounds were serially diluted 1/2 in 90% dmso to generate 7 point dose curves. 100nL of compound was dispensed directly from the compound plate into the wells of the assay plate to generate a dose curve starting at 20uM and ending at 313 nM. The assay plates were incubated at 37℃for 96 hours at 5% CO 2. On day 6, the assay plates were removed from the incubator and allowed to stand at room temperature for 30 minutes. Using a liquid processor, 20uL of CellTiter Glo reagent was added to each well. The assay plate was shaken for 2 minutes and allowed to rest on the bench for 10 to 15 minutes. Assay plates were read on a GloMax microplate reader using the CellTiter Glo protocol and data was analyzed using GraphPad Prism. The activity of various agents, including various stapled peptides in table E2, was determined. Some of the results are shown in table E1 below.
Table e1. As examples, certain data for various compositions.
Structural information and compositions of the stapled peptides are described in table E2.
1. Compound ID
2. Beta-catenin FP IC50 (nM):
A≤50nM；50nM＜B≤200nM；200nM＜C≤750nM；750nM＜D≤1000nM；E＞1000nM
3.NanoBRET Abs IC50(uM)：
A≤1.5uM；1.5uM＜B≤3.0uM；3.0uM＜C≤10.0uM；D＞10.0uM
4.DLD1 4％Abs IC50(uM)：
“+”≤1.0uM；1.0uM＜“++”≤5.0uM；“+++”＞5.0uM
colo320dm proliferation Abs IC50 (uM):
“+”≤10.0uM；10.0uM＜“++”≤20.0uM；“+++”＞20.0uM
6. calculated mass
7. Measured value m/z (positive mode)
8. Measured value m/z (negative mode)
9.C reduction of a double bond of C (e.g., -CH=CH-) to a single bond (e.g., -CH) 2 -CH 2 -). A: reduction of-ch=ch-to-CH in each staple-like structure 2 -CH 2 -; b: reduction of-ch=ch-to-CH in the C-terminal side staple-like structure 2 -CH 2 - (see, for example, see the preparation of I-428 and I-432 as examples).
Table e2 certain peptides and compositions thereof as examples.
Unless otherwise indicated, peptides are stapled (the present disclosure also provides, inter alia, non-stapled forms of such peptides, optionally protected by one or more protecting groups (e.g., protection of the N-terminus, C-terminus, side chain, etc.), and intermediates thereof). As understood by those skilled in the art, stapling can provide more than one stereoisomer (e.g., double bond E/Z and/or diastereoisomer). In some embodiments, the double bond in the staple-like structure is E. In some embodiments, the double bond in the staple-like structure is Z. In some embodiments, the isomers (or combinations thereof) are listed separately (typically based on reversed phase HPLC peaks in elution order (e.g., by UV (e.g., at 220 nm) and/or MS detection) in that each earlier eluting peak is assigned a smaller ID number than each later eluting peak (if any), in some cases the peak may comprise two or more isomers, in some cases the isomers are not separated (or a single isomer), e.g., when there is one peak on HPLC). The compositions used in the various assays are typically stapled peptides; the present disclosure also provides peptides and compositions thereof prior to stapling. General HPLC method: xselect CSH C18 column 1.7um 2.1X150 mm Column temperature 40 ℃; flow 0.6 mL/min; 0.1% formic acid in both acetonitrile and water, from 5% to 95% acetonitrileIs 7.2 minutes. In some embodiments, different gradients and/or C8 columns are used. />
Some of the results of various additional evaluations of various additional agents and compositions are shown in table E3 below. For illustration, see tables E1 and E2. These data particularly determine that the techniques of this disclosure may provide a variety of activities and/or benefits.
Table E3. is given as an example for certain data for various compositions.
For the agents described in the tables, as previously described, in various embodiments, the N-terminus is blocked (N-terminus) by R 1 Amino (R) to the first amino acid (AA 1) 1 ) And (5) connection. In some embodiments, the N-terminal end cap may be considered as part of AA1 as appropriate. From there, each carboxylate (R 2 ) Amino group (R) of subsequent amino acid 1 ) Linking until the carboxylate radical of the last amino acid (R 2 ) R with C-terminal group 1 And (5) connection. For any compounds having a branching point (R 3 ) And the branching monomers are indicated in brackets, R of the monomers in brackets 1 R to amino acids 3 And (5) connection. For a polymer having two potential branching points (R 3 And R is 4 ) If two branches are indicated, R of the first branch 1 And R is R 3 Linked, and R of the second branching 1 And R is R 4 And (5) connection. For any pair of amino acids ending in the 3 designation, the R of each of these amino acids 3 The groups are all linked to each other. Likewise, for any pair of amino acids ending in the nomenclature 3, R of these amino acids 3 The groups are all linked to each other. For any inclusion having R 3 Agents of branched amino acid pairs of groups and comprising a group comprising R 1 And R is R 2 Agents of branched monomers of both groups, R 1 To branched amino acids adjacent thereto in the sequence, and R of the branched monomer 2 R of groups with amino acids not specified as branching monomers 3 And (5) connection. For example, in various peptides having one of Cys, hCys, pen or aMeC at position 10 and one of Cys, hCys, pen or aMeC also at position 14, and a branching group other than the amino acid residue at position 10, R of the branching group 1 R to amino acid residue at position 10 3 To which R is attached, and the branching groups 2 R to amino acid residue at position 14 3 Are connected. For having a structure containing R 3 And not through any of the amino acids mentioned above attached thereto, R 3 =h. In various embodiments (e.g., the agents described in table E1, table E2, and table E3), pyrS2 is attached to one arm of R4, R5, R6, or B5, and if PL3 is present, it is typically attached to the other arm of B5. In various embodiments, if the N-terminal group comprises an olefin, it is attached to AA3 or a branching group of AA 3. If the peptide has been reduced as shown, then after olefin metathesis the olefin has been hydrogenated to-CH 2 -CH 2 -; if "C-only" is indicated, only the C-terminal side staple structure (e.g., in many cases, the pyrS2/R5 olefin staple structure) is hydrogenated to-CH 2 -CH 2 -. For non-hydrogenated peptides, each olefin metathesis can produce two possible staple structural isomers, yielding 2 n The possible isomers (four if n=2). For peptides having the same description and different designation numbers, the peptides are two separable isomers or a composition comprising one or more isomers. In various embodiments, for peptides comprising an amino acid residue beginning with "Dap7" or "DapAc7", the alkene of the amino acid residue is attached to one arm of B5 by alkene metathesis, while the R of the stapled amino acid residue 3 R of a group with another amino acid residue elsewhere in the peptide (e.g., glnR 3 residue) 3 Are connected. Special cases: for I-1484 and I-1485, PL3 is stapled to S5, while R5 residues are stapled to pyrS 2.
In some embodiments, it was determined that a variety of peptides (e.g., stapled peptides) comprising the amino acid residues described herein can provide higher affinity than a reference peptide comprising a reference amino acid (e.g., a natural amino acid such as Asp or Glu), but otherwise be identical.
Example 5 preparation of amino acids for peptide synthesis.
In some embodiments, the present disclosure provides a variety of compoundsAnd (3) an object. In some embodiments, such compounds may be used to incorporate related amino acids into peptides. In some embodiments, such compounds are compounds 2-2Or salts thereof, including methods, reagents, intermediates, and the like, are described in priority applications WO 2022/020651 or WO 2022/020652 and incorporated herein by reference.
Example 6 preparation of amino acids for peptide synthesis.
In some embodiments, the present disclosure provides a plurality of compounds. In some embodiments, such compounds may be used to incorporate related amino acids into peptides. In some embodiments, such compounds areOr a salt thereof, the preparation and use (including methods, reagents, intermediates, etc.) of which are described in priority applications WO 2022/020651 or WO 2022/020652, and incorporated herein by reference.
Example 7 preparation of amino acids for peptide synthesis.
In some embodiments, the present disclosure provides a plurality of compounds. In some embodiments, such compounds may be used to incorporate related amino acids into peptides. In some embodiments, such compounds are Or a salt thereof, the preparation and use (including methods, reagents, intermediates, etc.) of which are described in priority applications WO 2022/020651 or WO 2022/020652, and incorporated herein by reference. In some embodiments, the present disclosure provides a plurality of compounds. In some embodiments, such compounds areOr a salt thereof, its preparation and use (including methods, reagents, intermediates, etc.) in the priority application WO 2022/020651 or WO 2022/020652, and incorporated herein by reference.
Example 8. Additional examples of manufacturing techniques.
Compounds having a substitution at the 2-aminophenylalanine residue (e.g., I-1660 to I-1672) were synthesized in the following manner: ac-PL3-Asp-Npg-B5-Asp-3COOHF-Aib-Ala-Phe-Lys 3-pyrS2-2NO2F-BztA-GlnR 3-Ala-protide resin was synthesized on Liberty Blue as above and lactam cyclisation and olefin metathesis were performed as above. The nitro group was reduced by treatment with 30 equivalents of tin (II) chloride (2M solution in DMF) at 100 ℃ for 10 minutes. The resin was drained and washed with DMF. The resulting peptide was treated with the corresponding carboxylic acid (7 eq), HATU (7 eq) and diisopropylethylamine (14 eq) at 40 ℃ for 2 hours. The coupling reaction was repeated in the case of incomplete reaction. The resin was washed with DMF and dichloromethane and the peptide was cleaved and purified as above.
(R) -N-Fmoc-2- (2' -propenyl) alanine (Fmoc-R3-OH, CAS 288617-76-5) (10.0 g,30 mmol) was dissolved in dichloromethane (90 mL) and diisopropylethylamine (30.5 mL,180 mmol) and 2-chlorotrityl resin (28.1 g,30 mmol) were added. The resin was stirred at room temperature for 2 hours, and methanol (30 mL) was added and the resin was stirred for an additional 30 minutes. The resin was washed with DMF (3X 60 mL) and then treated with 20% piperidine in DMF (60 mL). The resin was stirred at room temperature for 30 min, then the resin was washed with DMF (4X 60 mL) and methanol (3X 60 mL). The resin was then treated with a mixture of hexafluoroisopropanol (18 mL) and dichloromethane (72 mL) and the mixture was stirred for 40 minutes. The resin was filtered off and the resulting solution was concentrated to give R3-OH.
R3-OH (7.88 g,55.5 mmol) was dissolved in methanol (100 mL) and thionyl chloride (13.2 g,111 mmol) was added at 0deg.C and the reaction was warmed to reflux and stirred for 14 hours. All volatiles were removed under vacuum to give the R3-OMe HCl salt (13.2 g) which was directly used in the next step.
To a solution of R3-OMe HCl salt (6.20 g,28.6 mmol) in THF (100 mL) and triethylamine (10.0 mmol,71.7 mmol) was added 4-bromobutyryl chloride (5.0 mL,43.0 mmol) at room temperature. The reaction was stirred at room temperature for 4 hours, then saturated ammonium chloride (100 mL) was added. The mixture was extracted with ethyl acetate (3×100 mL), and the combined organic layers were washed with 1M HCl (200 mL), brine (150 mL), dried over sodium sulfate and concentrated in vacuo. The residue was purified by silica gel chromatography (10% to 50% ethyl acetate in petroleum ether) to give 4-bromobutyric acid R3-OMe (3.90 g,13.3mmol,46.5% yield).
To a solution of R3-OMe (3.90 g,13.3 mmol) of 4-bromobutyric acid in THF (70 mL) was added sodium hydride (961 mg,24 mmol) and the reaction was stirred at room temperature for 3 hours. The mixture was diluted with ethyl acetate (20 mL) and quenched with saturated ammonium chloride (30 mL). The mixture was extracted with ethyl acetate (3×25 mL), and the combined organic layers were dried over sodium sulfate and concentrated. The remaining crude residue was purified by silica gel chromatography (20% to 50% ethyl acetate in petroleum ether) to give a yellow oil. The oil was dissolved in methanol (50 mL) and water (50 mL), and lithium hydroxide hydrate (1.27 g,30 mmol) was added. The reaction was stirred at room temperature for 1 hour. Methanol was removed under vacuum and the residue was extracted with ethyl acetate (30 mL). The aqueous layer was acidified to ph=3 with 1N HCl and extracted with dichloromethane (5×30 mL). The combined dichloromethane layers were concentrated under vacuum to give NPyroR3-OH (2.54 g,12.8mmol,96% yield).
To a solution of compound 1 (25.0 g,113 mmol) in THF (500 mL) was added potassium hydroxide (38.0 g,678 mmol) and propargyl bromide (101 g,678 mmol) in portions. The reaction was stirred at room temperature for 14 hours and the mixture was filtered and the filtrate concentrated in vacuo. Silica gel chromatography (1% to 10% ethyl acetate in petroleum ether) yielded compound 2 (23.2 g,69.2mmol,61% yield).
The mixture of 2 (23.2 g,69.2 mmol) was stirred in HCl solution (4M in ethyl acetate) for 30 min at room temperature. All volatiles were removed under vacuum to give compound 3 (18.4 g,67.7mmol,98% yield) as a pale yellow solid.
To a solution of PEG 4-diacid (7.74 g,26.3 mmol) in DMF (100 mL) was added HATU (10.0 g,26.3 mmol) and diisopropylethylamine (8.33 mL,47.8 mmol). The mixture was stirred at room temperature for 30 minutes, then compound 3 (6.5 g,23.9 mmol) was added. The reaction was stirred at room temperature for 2.5 hours, and the reaction was diluted with water (9500 mL) and extracted with ethyl acetate (3×200 mL). The combined organic layers were washed with brine (200 mL) and dried over sodium sulfate. The residue was purified by reverse phase HPLC to give compound 4 (3.5 g,6.84mmol,29% yield). LCMS M/z=512 (m+h).
I-1525, I-1526: at X 18 The compounds with branched PEG at them were synthesized in the following manner: synthesis by solid phase peptide Synthesis as above
Ac-PL3-Asp-Npg-B5-Asp-3COOHF-Aib-Ala-Phe-Lys 3-PyrS2-3Thi-BztA-GlnR 3-Ala-Ala-Lys (ivDde) -protide resin and lactam cyclisation and olefin metathesis were carried out as above. The ivDde groups were removed by treating the resin with 5% hydrazine in DMF at 40 ℃ for 30 min, and the resin was drained and washed with DMF. The resin was treated with compound 4 (3 eq), HATU (3 eq) and diisopropylethylamine (10 eq) at 40 ℃ for 3 hours. The peptide was cleaved as above and purified by reverse phase HPLC. The purified peptide (500 mg) was dissolved in 1:1 acetonitrile in water, and 4 equivalents of mPEG 16-azide (for I-1525) or mPEG36 (for I-1526) were dissolved in 1:1 acetonitrile in water and added to the peptide solution. The pH was adjusted to about 8 with ammonium bicarbonate, and copper sulfate (4 equivalents) and sodium ascorbate (5 equivalents) were added and again adjusted to about 8 with ammonium bicarbonate, if desired. The reaction was stirred at 40 ℃ for 2 hours and the final peptide was purified by preparative HPLC to give I-1525 (65% yield) or I-1526 (55% yield).
Example 9 the provided techniques may provide high selectivity.
The present disclosure provides, inter alia, various techniques for preparing stapled peptides, including those containing various staple-like structures. As described herein, in some embodiments, two or more staple-like structures are formed in one step. For example, in some embodiments, two or more staple-like structures are formed in the metathesis reaction. In some embodiments, all of the staple-like structures formed by metathesis are formed in the metathesis reaction. In some embodiments, each such staple-like structure is formed by olefin metathesis of a terminal olefin. In some embodiments, the plurality of staple-like structures are formed after the full length of the peptide has been reached. In some embodiments, one or more staple-like structures comprising double bonds are formed after the full length of the peptide has been reached. In some embodiments, all staple-like structures comprising double bonds are formed after the full length of the peptide has been reached. In some embodiments, the one or more staple-like structures formed by metathesis are formed after the full length of the peptide has been reached. In some embodiments, all of the staple-like structures formed by metathesis are formed after the full length of the peptide has been reached.
For example, in some embodiments, to prepare I-66 and I-67, the full-length peptide (in some embodiments, prepared on a solid phase, as shown below) is subjected to olefin metathesis:
in some embodiments, a ratio of about 3:1 (I-66:I-66) is observed.
In some embodiments, the staple-like structures are formed in two or more staple-like structures. In some embodiments, two or more olefin-containing staple-like structures are formed in the two or more staple-like structures. In some embodiments, two or more staple-like structures are formed in two or more metathesis steps. In some embodiments, two or more metathesis steps utilize different conditions, e.g., different catalysts. In some embodiments, each staple-like structure is formed in a separate step. In some embodiments, each staple-like structure comprising a double bond is formed in a separate step. In some embodiments, each staple-like structure comprising an olefin is formed in a separate step. In some embodiments, each staple-like structure formed by olefin metathesis is formed in a separate metathesis step. In some embodiments, progressive stapling provides improved levels of selectivity for formation of the desired product (e.g., I-66) as compared to other compounds (e.g., stereoisomers (e.g., for I-66, I-67)). For example, in some embodiments, I-66 is prepared as described below, and I-66:I-67 ratios in excess of 10:1 are observed. In some embodiments, the present disclosure provides compositions comprising I-66, wherein the ratio of I-66 to I-67 is about or at least about 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 25:1, 30:1, 40:1, 50:1, 60:1, 70:1, 80:1, 90:1, or 100:1. In some embodiments, the present disclosure provides compositions comprising I-66 and I-67, wherein the ratio of I-66 to I-67 is about or at least about 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 25:1, 30:1, 40:1, 50:1, 60:1, 70:1, 80:1, 90:1, or 100:1. In some embodiments, the ratio is about or at least about 5:1. In some embodiments, the ratio is about or at least about 10:1. In some embodiments, the ratio is about or at least about 20:1. In some embodiments, the ratio is about or at least about 30:1. In some embodiments, the ratio is about or at least about 50:1. In some embodiments, the ratio is about or at least about 80:1. In some embodiments, the ratio is about or at least about 90:1. In some embodiments, the ratio is about or at least about 100:1. In some embodiments, I-66 is provided in salt form (e.g., pharmaceutically acceptable salt form). In some embodiments, I-66 is provided in a variety of forms, including a variety of salt forms. In some embodiments, I-67 is provided in salt form (e.g., pharmaceutically acceptable salt form). In some embodiments, I-67 is provided in a variety of forms (including a variety of salt forms).
In the formulation, I-66 was synthesized by manual SPPS on Rinkamide MBHA resin (98 g,0.51mmol/g load, 50mmol total). The deprotection step was performed by treating the resin with 20% piperidine in DMF (v/v, 1000 mL) with agitation by nitrogen bubbling for 30 min. The resin was drained and washed four times with DMF. The amino acid to be coupled was dissolved in DMF (800 mL) and the coupling agents shown below and Diisopropylethylamine (DIEA) or HOAt were added in the equivalent amounts listed below. The coupling was performed at room temperature with nitrogen bubbling for 30 minutes and the amino acid solution was drained and the resin was washed four times with DMF.
After Aib addition, the resin was washed five times with DMF and five times with dichloromethane before Fmoc deprotection. A solution of phenylsilane (54 g,500 mmol) and tetrakis (triphenylphosphine) palladium (0) (5.77 g,5 mmol) in dichloromethane (500 mL) was added. The reaction was carried out at room temperature with nitrogen bubbling for 15 minutes and the palladium solution was discharged. The palladium/phenylsilane treatment was repeated two more times and then the resin was drained and washed five times with DMF. The lactam was blocked by bubbling the resin with HOAt (400 mmol) and DIC (400 mmol) in DMF (1000 mL) with nitrogen at room temperature for 2 hours. The resin was drained and washed four times with DMF. The cycles of Fmoc deprotection and amino acid addition were continued as above. The Fmoc-Npg-OH was subjected to repeated coupling steps.
Amino acids used Coupling agent/base and amount used
Fmoc-3COOHF(tBu)-OH(65mmol) HATU(61.5mmol)，DIEA(65mmol)
Fmoc-Asp(tBu)-OH(80mmol) HBTU (75 mmol) and DIEA (80 mmol)
Fmoc-B5-OH(65mmol) HATU (61.5 mmol and DIEA (65 mmol)
Fmoc-Npg-OH(75mmol)(x2) HATU (70 mmol) and DIEA (75 mmol) (x 2)
Fmoc-Asp(tBu)-OH(75mmol) HBTU (70 mmol) and DIEA (75 mmol)
After coupling Asp2, the B5/pyrS2 staple like structure was blocked by treating the resin with Hoveyda-Grubbs M720 catalyst (15.7 g,25 mmol) and 1, 4-benzoquinone (13.5 g,125 mmol) in dichloroethane. The reaction was bubbled with nitrogen at room temperature for 2 hours, the catalyst was vented and the treatment with M720 catalyst and 1, 4-benzoquinone was repeated once more, and then continued with linear peptide synthesis.
Amino acids/reagents used Coupling agent/base and amount used
Fmoc-PL3-OH(75mmol) HBTU(75mmol)，DIC(75mmol)
Ac2O(200mmol) DIEA(100mmol)
After N-terminal acetate capping, the PL3/B5 staple structure was blocked by bubbling the resin with Grubbs catalyst M102 (20.6 g,25 mmol) in dichloroethane for 2 hours at room temperature with nitrogen. The catalyst solution was drained and the treatment with Grubbs catalyst M102 was repeated two additional times. The peptide was cleaved by treating the resin with 95:5 TFA: water (800 mL, v/v) for 2 hours, and the peptide was precipitated by pouring the cleavage mixture into cold methyl tert-butyl ether. The precipitated peptide was filtered, washed twice with cold MTBE, and dried under vacuum. The peptide was first purified by dissolution in DMF and loaded to Luna C8 10um in 50 minutes The gradient was 45% to 75% acetonitrile in water (containing 0.075% TFA) on a column (flow: 20 mL/min). Drying the product-containing fraction and subjecting the isolated peptide toSecond purification and dissolution in 30% acetonitrile in water and loading into Kromasil C8 5 μm->On the column (20 mL/min), 0.4M ammonium acetate was first passed through the column for 25 min, followed by a gradient elution with 50% to 70% acetonitrile in water (containing 0.5% acetic acid) over 50 min. The product-containing fractions were lyophilized to provide I-66 (40:1I-66:I-67, 4997mg,2.41mmol,4.8% yield) plus a second crop of I-66 (8:1I-66:I-67, 2015mg,0.97mmol,1.9% yield). The ratio of I-66 to I-67 was evaluated using HPLC: agilent Poroshell 120 EC-C18; 4.6X100 mm; solvent a = 0.1% tfa in water; solvent B = 0.075% tfa in acetonitrile; gradient 10% b to 95% b over 30 minutes; the detection is UV absorbance at 220 nM; and calculates the ratio based on the peak area. As an example, in one run, the retention time of I-66 was 15.3 minutes and the retention time of I-67 was 16.2 minutes. In some embodiments, such schemes provide improved resolution compared to reference schemes by which I-66 and I-67 may be eluted as one peak or otherwise insufficiently separable. For example, by the general procedure used in Table E2, I-66 and I-67 may be eluted together as a second peak, and the mixture may be named I-67). Alternatively or additionally, other techniques (e.g., NMR) may also be used to evaluate the ratio. In some embodiments, such I-66 formulations, or formulations corresponding thereto, are evaluated in a variety of bioassays and are determined to have a variety of properties and activities; see, for example, examples 11 to 18. Such I-66 formulations 1 H NMR is shown in fig. 6. Those skilled in the art who review this disclosure will appreciate that FIG. 6 may contain some impurities and/or residues in the NMR solvent 1 A peak of H. In some embodiments, NOE is observed between about 5.45 to 5.6 and peaks at about 5.2 to 5.35. The fractions may be further purified to provide increased purity.
In some embodiments, I-66 and/or I-67 prepared herein can be used as a standard/reference to evaluate and/or characterize other compounds and/or other formulations of I-66 and/or I-67 (e.g., different batches prepared by the same or different methods). In some embodiments, I-470 is similarly prepared. In some embodiments, I-470 differs from I-66 in that I-470 has Glu2 and Glu5, and I-66 has Asp2 and Asp5.
In some embodiments, the present disclosure provides a composition havingA compound of the structure or a salt thereof. In some embodiments, the present disclosure provides a composition having ∈>A compound of the structure or a salt thereof. In some embodiments, the present disclosure provides a composition having ∈>A compound of the structure or a salt thereof. In some embodiments, the compound has the same retention time as I-66 prepared above under the same or comparable HPLC conditions. For example, in some embodiments, the HPLC conditions are Agilent Poroshell EC-C18; 4.6X100 mm; solvent a = 0.1% TFA in water; solvent B = 0.075% TFA in acetonitrile; gradient 10% B to 95% B in 30 minutes; the detection is UV absorbance at 220 nM; and the retention time of I-66 was about 15.3 minutes. In some embodiments, HPLC conditions separate I-66 and I-67. In some embodiments, the compound elutes as a single peak of I-66 when co-injected with an I-66 formulation described herein. In some embodiments, the compound is characterized in that it is 1 In the H NMR spectrum, it shows peaks overlapping with peaks between about 5.1 and 5.7 in fig. 6. In some embodiments, the compound is characterized in that it is 1 In the H NMR spectrum, it has the same peak pattern as in fig. 6 between about 5.1 and 5.7. In some embodiments, the compound has the same NMR spectrum as I-66 under the same or equivalent conditions. In some embodiments, the compounds have the same I-66 under the same or equivalent conditions (e.g., DMSO-d6, 373K) 1 H NMR spectrum.Those skilled in the art will appreciate that for the same compound, certain of the following are during different evaluations 1 Peaks of H (e.g., those bound to nitrogen and oxygen) may be present 1 Offset in H NMR. In some embodiments, the compound has a structure that, under the same or equivalent conditions (DMSO-d 6, 373K), is 1 H NMR, carbon-bonded 1 The peak of H is shown in fig. 6. In some embodiments, the compound has a structure that, under the same or equivalent conditions (DMSO-d 6, 373K), is 1 HNMR peaks are shown in fig. 6. In some embodiments, the compound has a structure that is bonded to a carbon atom under the same or equivalent conditions (DMSO-d 6, 373K) 1 H is its 1 The H NMR peaks are shown in fig. 6. In some embodiments, each of the compounds in FIG. 6 corresponds to a carbon-bonded group when the integral of the triplet state at about 5.45 to about 5.6 is set to 1 1 The integral of the peaks of H is independently about 1 (e.g., about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 to about 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9 or 2.0, about 0.2 to 1.8, about 0.5 to 1.5, about 0.7 to 1.5, 0.8 to 1.2, etc.). In some embodiments, each of the compounds corresponds to in fig. 6 when the integral of the triplet state at about 5.45 to about 5.6 is set to 1 1 The integral of the peaks of H is independently about 1 (e.g., about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 to about 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9 or 2.0, about 0.2 to 1.8, about 0.5 to 1.5, about 0.7 to 1.5, 0.8 to 1.2, etc.). As an example, the integration of fig. 6 is shown in fig. 7. Those skilled in the art will understand 1 H NMR results (e.g., chemical shifts, peak integration, etc.) can have typical error ranges. In some embodiments, corresponding to two or more 1 The peaks of H may overlap. In some embodiments, such peaks may be integrated together for evaluation 1 Number of H. In some embodiments, the NMR of the above-described I-66 formulation is the same or equivalent, with or without the addition of a detectable level of a compound (e.g., the same amount of I-66), under the same or equivalent conditions. In some embodiments, with or without the addition of a detectable agentMeasuring the level of a compound (e.g., I-66 or about 0.1 to 10, 0.2 to 5, 0.5 to 2, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 2, 3, 4, 5, 6, 7, 8, 9 or 10, etc.) of I-66 in the same amount 1 H NMR is the same or equivalent. In some embodiments, when corresponding to bonding to carbon 1 Where the peak of H has a comparable chemical shift, peak shape and/or integral, 1 h NMR is considered to be the same or equivalent. In some embodiments, peaks from impurities and solvent are properly excluded when comparing NMR. In some embodiments, from impurities, solvents, oxygen, nitrogen-bonded 1 The peak of H, etc. is appropriately excluded when comparing NMR. In some embodiments, the compound has the same retention time as I-67 prepared above under the same or comparable HPLC conditions. For example, in some embodiments, the HPLC conditions are Agilent Poroshell EC-C18; 4.6X100 mm; solvent a = 0.1% TFA in water; solvent B = 0.075% TFA in acetonitrile; gradient 10% B to 95% B in 30 minutes; the detection is UV absorbance at 220 nM; and a retention time of I-67 of about 16.2 minutes. In some embodiments, HPLC conditions separate I-66 and I-67. In some embodiments, the compound elutes as a single peak of I-67 when co-injected with an I-67 formulation described herein. In some embodiments, the compound has the same I-67 as between about 5.0 and 6.0 under the same or equivalent conditions 1 H NMR peaks. In some embodiments, the compound has the same NMR spectrum as I-67 under the same or equivalent conditions. In some embodiments, the compound has the same as I-67 under the same or equivalent conditions (e.g., DMSO-d6, 373K) 1 H NMR spectrum. In some embodiments, the NMR of the above-described I-67 formulation is the same or equivalent, with or without the addition of a detectable level of a compound (e.g., the same amount of I-67), under the same or equivalent conditions. In some embodiments, the I-67 formulation described above is formulated with or without the addition of a detectable level of a compound (e.g., the same amount of I-67) 1 H NMR is the same or equivalent. In some embodiments, the composition comprisesIn a composition of compounds, the ratio of the compound to the stereoisomer of the compound is about or at least about 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 25:1, 30:1, 40:1, 50:1, 60:1, 70:1, 80:1, 90:1, or 100:1. In some embodiments, in a composition comprising a compound, the ratio of the compound to each stereoisomer of the compound is independently about or at least about 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 25:1, 30:1, 40:1, 50:1, 60:1, 70:1, 80:1, 90:1, or 100:1. In some embodiments, the ratio of all compounds that are compounds or salts thereof to all compounds that are stereoisomers of the compounds or salts of the stereoisomers in a composition comprising the compounds is about or at least about 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 25:1, 30:1, 40:1, 50:1, 60:1, 70:1, 80:1, 90:1, or 100:1. In some embodiments, the ratio of all compounds that are the compounds or salts thereof to all compounds that are stereoisomers of the compounds or salts thereof in a composition comprising the compounds is about or at least about 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 25:1, 30:1, 40:1, 50:1, 60:1, 70:1, 80:1, 90:1, or 100:1 for each stereoisomer of the compounds. In some embodiments, the ratio is about or at least about 2:1. In some embodiments, the ratio is about or at least about 3:1. In some embodiments, the ratio is about or at least about 4:1. In some embodiments, the ratio is about or at least about 5:1. In some embodiments, the ratio is about or at least about 10:1. In some embodiments, the ratio is about or at least about 20:1. In some embodiments, the ratio is about or at least about 30:1. In some embodiments, the ratio is about or at least about 50:1. In some embodiments, the ratio is about or at least about 80:1. In some embodiments, the ratio is about or at least about 90:1. In some embodiments, the ratio is about or at least about 100:1.
In some embodiments, the I-66 formulation comprisesOr a salt thereof. In some embodiments, the I-67 formulation comprises +.>Or a salt thereof. In some embodiments, the I-66 formulation or I-67 formulation comprises the first compound +.>Or a salt thereof and a second compound +>Or a salt thereof. In some embodiments, the ratio of the first compound to the second compound in the I-66 formulation is about or at least about 2:1, 3:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 25:1, 30:1, 40:1, 50:1, 60:1, 70:1, 80:1, 90:1, or 100:1. In some embodiments, the ratio of all compounds that are the first compound or salt thereof to all compounds that are the second compound or salt thereof in the I-66 formulation is about or at least about 2:1, 3:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 25:1, 30:1, 40:1, 50:1, 60:1, 70:1, 80:1, 90:1, or 100:1. In some embodiments, the ratio of the second compound to the first compound in the I-67 formulation is about or at least about 2:1, 3:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 25:1, 30:1, 40:1, 50:1, 60:1, 70:1, 80:1, 90:1, or 100:1. In some embodiments, the ratio of all compounds that are the second compound or salt thereof to all compounds that are the first compound or salt thereof in the I-67 formulation is about or at least about 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 25:1, 30:1, 40:1, 50:1, 60:1, 70:1, 80:1, 90:1, or 100:1. In some embodiments, the ratio is about or at least about 2:1. In some embodiments, the ratio is about or at least about 3:1. In some embodiments, the ratio is about or at least about 4:1. In some embodiments, the ratio is about or at least about 5:1. In some implementations In embodiments, the ratio is about or at least about 10:1. In some embodiments, the ratio is about or at least about 20:1. In some embodiments, the ratio is about or at least about 30:1. In some embodiments, the ratio is about or at least about 50:1. In some embodiments, the ratio is about or at least about 80:1. In some embodiments, the ratio is about or at least about 90:1. In some embodiments, the ratio is about or at least about 100:1. As used in the present disclosure, depending on the context, in some embodiments, the ratio is a molar ratio; in some embodiments, the ratio is a weight ratio; in some embodiments, the ratio is a volume ratio; and in some embodiments, the ratio is based on the evaluation. For example, in some embodiments, when the ratio of compounds is assessed using HPLC/UV, the ratio is the peak area of the UV trace at a certain wavelength (e.g., 220 nm).
Example 10 the techniques provided may provide high selectivity.
As determined below, in some embodiments, the present disclosure provides techniques for forming staple-like structures comprising olefinic double bonds with high selectivity.
Fmoc-azido lysine-pyrS 2-3 Thi-BztA-propargylglycine-Ala-protide resin was synthesized using standard solid phase peptide synthesis procedures. The triazole staple structure was blocked by treating the resin with one equivalent of copper (I) iodide, one equivalent of sodium ascorbate, ten equivalents of diisopropylethylamine and ten equivalents of 2, 6-lutidine in dichloromethane for 48 hours at room temperature. Resin was washed with DCM 2X, meOH 1X, H 2 O 2×、50％ H 2 O/MeOH 2X and MeOH 2X washes for 5 minutes. In some embodiments, a small layer of insoluble material was observed to float on top of the reactor, which was removed by suction through a hose connected to a pump. Then, washing with NMP 2×, DCM 1×, and MeOH 1× was continued.
The cyclized product was extended to Fmoc-Asp (OtBu) -Npg-B5-Asp (OtBu) -3COOHF (OtBu) -Aib-Ala-Phe-TriAzLys 3-pyrS2-3Thi-Bzta-sA 3-Ala-protide resin using standard solid phase peptide synthesis procedures. After that, the resin was thoroughly washed with DCM 2×, NMP 1×, DCM 2×, meOH 2×, DCM 1×, meOH 1×, 5 minutes each time, and then dried under a nitrogen stream for 24 hours to give a gold resin. The first staple structure was blocked by treating the resin with 5mol% Hoveyda-Grubbs M720 catalyst (CAS 301224-40-8) and 10mol% benzoquinone in methylene chloride at reflux for 48 hours. After 48 hours, the catalyst solution was drained off, the resin was washed 3X with dichloromethane, dried, and then treated again with 5mol% Hoveyda-Grubbs M720 catalyst (CAS 301224-40-8) and 10mol% benzoquinone in dichloromethane under reflux for 48 hours.
After analysis by LCMS, a complete reaction was observed with no identifiable starting material. The desired product was detected > 95% and no other isomeric byproducts were observed. The double bond configuration is specified based on NMR data analysis, reported selectivity, etc., and can also be assessed by other techniques (e.g., crystallography).
The above product was extended to Ac-PL3-Asp (OtBu) -Npg-B5-Asp (OtBu) -3COOHF (OtBu) -Aib-Ala-Phe-TriAzLys 3-pyrS2-3Thi-Bzta-sA 3-Ala-protide resin using standard solid phase peptide synthesis procedures. After N-terminal acetyl capping, the resin was thoroughly washed with DCM 2×, NMP 1×, DCM 2×, meOH 2×, DCM 1×, meOH 1×, 5 minutes each, then dried under a nitrogen stream for 24 hours. The second staple structure was blocked by treating the resin with 30mol%Grubbs I M102 (CAS 172222-30-9) and 60mol% benzoquinone in methylene chloride at reflux for 24 hours. After 24 hours, the catalyst solution was drained off, the resin was washed 3X with dichloromethane, dried, and then treated again with 30mol%Grubbs I M102 in dichloromethane (CAS 172222-30-9) and 60mol% benzoquinone at reflux for 24 hours. The crude product was cleaved and deprotected and analyzed by LCMS and showed I-335 to be 82% (UV, e.g., at 210 to 400 nm). More than two peaks of olefin isomers were detected by HPLC at 13% and 5% of the total area, respectively. The double bond configuration is specified based on NMR data analysis, reported selectivity, etc., and can also be assessed by other techniques (e.g., crystallography).
Example 11 the techniques provided may provide a variety of advantages.
The techniques provided may provide a number of advantages, among others. In some embodiments, the provided techniques may provide improved target binding and/or activity profiles. As determined below, stapled peptides (particularly I-66) can provide strong binding to β -catenin and modulation of gene expression. Available schemes for various evaluations are described in the examples.
Peptide a: stapled peptides
Ac-PL3-Asp-Npg-B5-Asp-3COOHF-Ala-Ala-Phe-Leu-pyrS2-2F3MeF-BztA-Gln-NH2.PL3 and B5, and B5 and PyrS2 are stapled.
* : for I-66, T 1/2 =12.6 minutes.
* *: reduction of AXIN2 transcripts after administration of agents to COLO320DM cells.
In some embodiments, the provided technology (e.g., I-66) can inhibit the binding of TCF/LEF transcription factors to β -catenin, as determined, for example, by biochemical competition assays. In some embodiments, the provided technology (e.g., I-66) is observed to compete for β -catenin interactions with TCF1, TCF3, TCF4, LEF1, pAPC, mouse ECAD, human ECAD, and the like. In some embodiments, it was determined that the provided techniques (e.g., I-66) can significantly reduce phosphate-APC binding. In some embodiments, it was determined that the provided techniques (e.g., I-66) can significantly reduce E-cadherin binding. In some embodiments, little competition is observed for certain provided techniques (e.g., I-66) for ICAT, axin, or Bcl 9. In some embodiments, the interaction is dependent on phosphorylation, e.g., binding of E-cadherin to β -catenin is reported to be highly dependent on phosphorylation of up to 8 Ser residues on E-cadherin.
In some embodiments, the ability of the provided technology, e.g., to bind to β -catenin and/or disrupt its interaction (or lack thereof) with multiple partners, is assessed and determined in cells, e.g., in HEK293 cells using NanoBRET-based assays. In some embodiments, it is observed that the provided techniques (e.g., I-66) can potently inhibit such interactions without affecting cell viability.
Direct inhibition of endogenous β -catenin/TCF interactions was determined inter alia by co-immunoprecipitation (co-IP) assay as described herein.
Example 12 the techniques provided may regulate transcription.
The present disclosure particularly identifies that the provided technology can inhibit transcription of endogenous Wnt pathway target genes driven by β -catenin/TCF interactions. In particular, it was determined that peptides A and I-66 inhibited the expression of two true downstream genes AXIN2 and SP5 of β -catenin/TCF in DLD1 cells in a dose-dependent manner (peptide A: AXIN2 IC 50 ＝9.3uM，SP5 IC 50 ＝9uM；I-66：AXIN2 IC 50 ＝1.6uM，SP5 IC 50 =1.3 uM). In some embodiments, no effect on CTNNB1 expression of peptides a and I-66 in DLD1 cells was observed under the test conditions. A decrease in the expression level of the standard β -catenin target AXIN2 was also observed in COLO320DM cells (peptide A: IC 50 ＝1.4uM；I-66：IC 50 =0.3 uM), whereas I-470 has little or no significant effect.
The provided technology can particularly regulate transcription and levels of a variety of transcripts, in some embodiments, with certain types and/or levels of selectivity. For example, in a number of systems (e.g., HAP1 isogenic line (+/-CTNNB 1 knockdown)), the provided techniques can selectively modulate the expression level and/or activity of a nucleic acid (e.g., a gene, transcript, polypeptide, and/or product thereof) in a system comprising or expressing β -catenin. For example, in some embodiments, the provided technology selectively inhibits β -catenin driven transcription in HAP1 WT cells. Some data is shown as an example in fig. 1. In WT beta-catenin (WT) expressing cells, 24 hours CHIR treatment increased beta-catenin levels by more than twice, and peptide a and I-66 treatment significantly reduced the expression of AXIN2 and SP5 as measured by qPCR (to about one third and one eighth by I-66). Inhibition of RNF43 expression by peptides A and I-66 was also observed. In beta-catenin KO cells, neither CHIR nor peptides A and I-66 affected AXIN2, SP5 or RNF43 expression. No reduction in transcription of I-470 in WT cells was observed. In some embodiments, it is observed by western blotting that treatment with the provided peptide (e.g., 10um I-66) for 72 or 144 hours does not significantly affect β -catenin stability in cells with functional β -catenin disruption complexes.
Example 13 the techniques provided can reduce β -catenin levels in the nucleus.
In some embodiments, the provided techniques can reduce β -catenin levels in the nucleus. In some embodiments, the provided techniques may block β -catenin nuclear localization. In some embodiments, the provided techniques may reduce the level of beta-catenin nuclear translocation. For example, as determined in fig. 2, the provided techniques may reduce the level of nuclear β -catenin in a variety of cells, including COLO320DM cells (10 um,24 hours). The decrease in nuclear localization was also determined by immunofluorescence imaging. In some embodiments, a reduction in nuclear β -catenin levels of more than 70% is observed after 24 hours of I-66 treatment as compared to untreated cells. Similar results were obtained after 24 hours and 48 hours of treatment.
Example 14 the techniques provided can inhibit proliferation and induce cell cycle arrest.
As described herein, the provided technology can inhibit proliferation of a variety of cells, including a variety of cancer cells, among others. In some embodiments, provided techniques modulate WNT specific transcription. In some embodiments, the provided techniques induce cell cycle arrest. In some embodiments, the provided techniques induce G1 cell cycle arrest. In some embodiments, the provided techniques increase the proportion of cells in the G1 phase of the cell cycle. As determined in fig. 3, the provided techniques can inhibit proliferation of COLO320DM, which is a colorectal cell line that contains multiple mutations (e.g., APC, TP53, etc.), modulate WNT-specific transcription (e.g., AXIN2 and CXCL 12), and induce G1 cell cycle arrest. In some embodiments, it is determined that the provided techniques can reduce the proportion of cells in the S phase of the cell cycle. In some embodiments, it was determined that the provided techniques can significantly down-regulate cyclin D2 and up-regulate p27. In some embodiments, changes in various genes (e.g., AXIN2, CXCL12, etc.) are observed to be consistent with changes in shRNA knockouts. Two separate doxycycline (dox) inducible shRNA were used to Knock Down (KD) CTNNB1 in COLO320DM cells. A decrease in AXIN2 expression was observed, and an increase in CXCL12 expression was observed. CTNNB1-KD also significantly reduced proliferation of COLO320DM cells.
In some embodiments, the evaluation is performed as follows. On day 0, cells were seeded at the desired density (typically 1000 cells/well) in cell culture medium (RPMI 1640,4% FBS) in 96-well plates. On day 1, a 10mM stock solution of agent (in DMSO) was first serially diluted in DMSO at a ratio of 1:2, followed by dilution with twice the final concentration of cell culture medium. Finally, the medium containing the agent is introduced into the cell culture well already having the same volume of cell culture medium. Incubating the cells with the agent for a desired number of days and then lysing for useDetermination of luminous cell viability (according to the manufacturer's instructions (Promega, G7570)). Luminescence signals were obtained from a microplate reader (GloMax, promega). Cell viability data are expressed as% relative to DMSO control wells.
Example 15 the provided technology can provide robust anti-tumor effects in vivo.
As described herein, the provided techniques may be used to treat a variety of conditions, disorders, or diseases, including cancer. The present examples particularly confirm that the provided techniques can provide in vivo efficacy, as demonstrated in a variety of animal models. Some useful models and/or schemes are described below as examples. Those of skill in the art will appreciate from this disclosure that a variety of models of a variety of cancers may be utilized in accordance with this disclosure to evaluate the provided technology and determine its effect.
COLO320DM human colorectal cancer cells (ATCC, CCL-220) (which contain various mutations such as APC and TP53, etc.) were expanded in RPMI 1640 medium (10% FBS) and at 10 in a 100uLPBS/Matrigel (1:1) mixture 7 Individual cells/animals were inoculated subcutaneously into male NU/J mice (JAX # 2019) that were 8 weeks old. When the average tumor size reaches 150mm 3 At this time, mice were randomly divided into 3 groups (n=10) and treated with vehicle (1% tween 80/99%10mM PBS pH 7.4), I-66 (30 mg/kg) and I-66 (75 mg/kg) by intraperitoneal injection, once every 4 days for a total of 5 doses.
Tumor volume was measured by electronic calipers every 2 to 3 days until tumor volume reached 2000mm 3 And is estimated as (length×width 2 )/2. The body weight was weighed every 2 to 3 days and expressed as% body weight = (BWi-BW 0)/BW 0 x 100% (BWi: body weight at day i, BW0: body weight at day 0). Tumor growth inhibition was calculated as: TGI% = [1- (TVi-TV 0)/(TVvi-TVv 0)]X 100% (TVi: average tumor volume of the group administered at day i, TV0: average tumor volume of the group administered at day 0, TVI: average tumor volume of the group vehicle at day i, TVv0: average tumor volume of the group vehicle at day 0). On the designated end day of each study, by CO 2 The animals were euthanized by asphyxiation and plasma, tumors, tissues, etc. were removed for further analysis. Some data is shown as an example in fig. 4.
As determined, the techniques of the present disclosure may provide robust anti-tumor efficacy. For example, in some embodiments, I-66 was administered once every four days in the COLO320DM xenograft model, and the treatment resulted in significant tumor growth inhibition of 66% and 89% at 30 and 75mg/kg on day 14, respectively (tumor growth inhibition, TGI). At 75mg/kg, after the first administration, a weight loss was observed, but recovered over time.
In some embodiments, transcription of in vivo pathway inhibition is assessed. For example, in some embodiments, several PD markers of COLO320DM tumors obtained at the end of the efficacy study (e.g., day 18) were evaluated. Consistent with in vitro and single dose in vivo data, in tumors, both AXIN2 and CXCL12 were dose-dependently regulated (down-regulated for AXIN2 and up-regulated for CXCL 12) by the provided techniques (e.g., I-66), confirming durable target gene regulation. A decrease in mouse NOTUM levels in plasma was also observed. In some embodiments, NOTUM can be used as a biomarker, e.g., for evaluating treatment, selecting patient populations, determining whether to continue treatment, and the like. In some embodiments, evaluation of human plasma samples from normal humans and patients (e.g., colorectal cancer patients) determines that NOTUM levels are associated with stages of disease, and may be suitable for clinical applications, e.g., as target-engaging biomarkers.
Example 16. The provided technology can be delivered in vivo.
In particular, a number of suitable in vivo pharmacokinetic and/or pharmacodynamic properties and/or activities are determined. For example, as determined in fig. 5 (a), the provided techniques may be effectively delivered to a tumor. As shown, prolonged tumor exposure to I-66 was observed after a single dose, and tumor exposure was about 2 to 10 times higher than in vitro proliferation IC50. I-66 tumor PK was also observed to exceed plasma PK at the 96 hour time point. Furthermore, I-66 provides a significantly longer period of time than, for example, peptide a, during which the tumor is exposed to about or above the in vitro proliferation IC50.
The experiments were conducted under the protocol approved by the institutional animal care and use committee (Institutional Animal Care and Use Committee) and followed institutional guidelines for proper and humane use of animals.
For COLO320DM, male NU/J mice (6 to 8 weeks old) were used and when the average tumor volume reached 300mm 3 Mice were randomized at this time. For IP administration, the formulation was formulated as 10mg/mL arginine and6% PEG400 phosphate (pH 7.4) formulation.
The concentration of the agent in the biological sample was measured by LC-MS/MS (triple quadrupole 6500+). Using analytical grade chemicals and solvents, 25ng/ml tolbutamide in acetonitrile (ACN, LS120-4,Fisher Scientific) was used as an internal standard. 8. Mu.L of plasma or tissue lysates were used in LC method, mobile phase A (H 2 1% formic acid in O (FA, LS118-4,Fisher Scientific)) and mobile phase B (0.1% FA in ACN), flow was 0.6 ml/min in a Waters ACQUITY UPLC BEH C18.1 x 50mm,1.7 μm column. Calibration curves were generated using 5 to 5000ng/mL of agent (e.g., I-66) in mouse plasma and tissue homogenates. MS was performed by electrospray ionization and multiple reaction monitoring scans. PK parameters such as plasma maximum concentration (Cmax) and AUC were analyzed by the non-compartmental model 200 of Phoenix WinNonlin 8.3.3 using a linear/logarithmic trapezoidal method.
Additional data confirm the good Pharmacokinetic (PK) profile of the provided technology. See, for example, fig. 5 (B) and the following data.
Certain PK parameters of I-66 in mice were analyzed by the 2-compartment model.
PK parameters IV IP
Cmax(ng/mL) 498654 152000
T 1/2 (hours) 28.71 41.7
T max (hours) NA 6.67
Vd ss (L/kg) 0.452 NA
Cl (mL/min/kg) 0.30 NA
T last (hours) 168.00 168.00
AUC 0-last (ng, hours/mL) 2741991 2971069
AUC 0-inf (ng, hours/mL) 2826191 3124730
Bioavailability (%) NA 105.0
In some embodiments, a broad tissue distribution is observed. For example, as shown in FIG. 5, (C), I-66 was detected in all of the samples shown. In some embodiments, persistent tissue residence is determined at least between 24 and 96 hours after injection. In some embodiments, a single Intraperitoneal (IP) dose of 100mg/kg of I-66 and I-470 exhibited comparable plasma AUC in mice.
Robust and durable anti-tumor effects of the provided technology were determined in additional tumor models. In some embodiments, such effects are observed in a Patient-derived xenograft (PDX) cancer model. In some embodiments, the model is a mouse PDX colon cancer model. In some embodiments, the model has APC mutation (Tyr 935Ter His1490LeufsTer 20) and high AXIN2 expression. In some embodiments, the LogCPM is about 2.5 or greater for AXIN2 expression. In particular, strong antitumor activity and durable tumor growth inhibition were determined. For example, 50mg/kg of animals dosed were observed to have tgi=103% at day 45. No significant weight loss was observed. As an example, some data is shown in fig. 8 (a). In some embodiments, the provided techniques are evaluated in a mouse model carrying a patient-derived xenograft colorectal tumor. Also, robust anti-tumor effects were determined. As an example, some data is shown in fig. 8 (B). Vehicle vs. i-66, p=0.008. Model mutation characterization: APC mutant, KRAS WT. IP dosing Q4D, n=10/group. In some embodiments, the animal is administered and/or observed for a longer period of time, e.g., more than 24 days. No significant weight loss was observed. For both PDX evaluations, the vehicle was 10mM disodium hydrogen phosphate, 6% w/w PEG-400, 10mg/mL L-arginine.
The data in the various embodiments determine, among other things, that the provided techniques can provide robust PK properties, strong anti-tumor efficacy, and targeted transcriptional regulation in vivo.
Example 17 the provided technology modulates expression in vivo.
As described herein, provided techniques can modulate the expression of a variety of nucleic acids and/or the level of products thereof (e.g., RNA transcripts, polypeptides, etc.). For example, tumor RNA sequencing analysis determined that I-66 can provide strong targeted Wnt/β -catenin pathway modulation, particularly in COLO320DM tumors. Some negatively enriched gene sets are shown below as examples. In some embodiments, the negative enrichment gene is CCND2, WNT5B, AXIN, NKD1, WNT6, DKK1, or DKK4. Notably, both negative and positive enriched gene sets were observed. The present disclosure provides, inter alia, techniques for assessing the efficacy of a method (e.g., a treatment) comprising assessing the expression of one or more negatively and/or positively enriched genes. In some embodiments, if the expression profile of one or more genes is negatively and/or positively enriched as identified herein, the method can be considered to be effective, and/or administration (e.g., provided techniques, such as stapled peptides, compositions, etc.) to a subject can continue.
The pre-negative enrichment gene set comprises
BCAT_GDS748_UP，BCAT.100_UP.V1_UP，HALLMARK_WNT_BETA_CATENIN_SIGNALING，RASHI_RESPONSE_TO_IONIZING_RADIATION_1，REACTOME_RRNA_PROCESSING，HALLMARK_MYC_TARGETS_V1，HALLMARK_MYC_TARGETS_V2，HALLMARK_OXIDATIVE_PHOSPHORYLATION，HALLMARK_E2F_TARGETS，HALLMARK_TNFA_SIGNALING_VIA_NFKB.
I-66 vs. I-470. P.30mg/kg, 48 hours after a single dose. NES-1.7 or less. FDR q value is 0.02 or less.
At 48 hours post-dose, comparable concentrations of I-66 and I-470 (e.g., 4266 and 5181ng/g, respectively, in the evaluation) were determined in the tumor. As determined, GSEA revealed multiple Wnt/β -catenin and MYC-related gene sets ordered as top hits in the negative-enriched gene set. Consistent with the cell-based data, the results confirm that the provided techniques (e.g., I-66) can provide strong targeted Wnt/β -catenin pathway modulation in tumors, as shown herein in COLO320DM tumors.
In some embodiments, the present disclosure provides techniques for identifying regulated nucleic acids and/or products thereof (including gene sets) and how they are regulated. In some embodiments, one or more nucleic acids and/or products thereof, or a set (e.g., a gene set) of nucleic acids and/or products thereof, modulation patterns can be used to select a patient population for treatment or to continue or adjust treatment (e.g., dose levels, regimens, etc.).
Usable schemes are described below as examples.
RNAseq preparation. For RNA-seq of cell line engrafting tumors, library preparation and sequencing is performed with a suitable kit (e.g., truSeq strand mRNA library kit on the Novaseq S4 platform), in some embodiments enriched with PolyA.
RNAseq data analysis. In some embodiments, trimmings of sequence are used to remove possible aptamer sequences and poor quality nucleotides. The trimmed reads were mapped to the homo sapiens GRCh38 reference genome available on ENSEMBL using STAR aligner v.2.7.7a. For transplanted tumor samples, host reads were removed with XenofilteR. Unique gene hit counts were calculated by using featureCounts in the R test package v.2.4.2. Readout filtration, normalization and differential expression analysis were performed with the edge package v.4.0.2 in R. For each comparison, genes with adjusted p-values < 0.01 and absolute log2 fold change > 1 were referred to as differentially expressed genes. Genes differentially expressed in at least one comparison were used for heat map and cluster analysis. Gene expression was normalized to fold change compared to a simultaneous reference (e.g., DMSO) control. The R pheeatmap package v.1.0.12 was used to make heat maps and hierarchical clustering of genes with relevance as a similarity measure. For enrichment analysis, GSEA v4.1.0 was run with a fold-change gene list in MSigDB database v 7.3. In some embodiments, a Venn plot is generated with ggvenn v.0.1.9, wherein overlapping p-values are calculated using the hypergeometric test in rv4.1.2. Those skilled in the art will appreciate that other software, programs, and/or algorithms may be utilized.
The time and dose dependent effect of the provided technology (e.g., I-66) on expression was also observed in COLO320DM cells by RNA seq. It was determined that treatment by the provided techniques (e.g., I-66) resulted in both time-and dose-dependent effects on the COLO320DM transcriptional profile. In some embodiments, at 1uM, 0, 107, and 359 differentially expressed genes (differentially expressed gene, DEG) were detected at 6 hours, 24 hours, and 48 hours, respectively, after treatment. At 10uM 73, 876 and 1271 DEG were found at three time points, respectively. RNAseq data from shRNA expressing cells after 3 days dox treatment was also assessed. In some embodiments, CTNNB1-KD produced by shRNA and provided techniques (e.g., I-66) was observed to result in consistent transcriptome changes in COLO320DM (r2=0.68, p < 2.2E-16).
To assess the impact of the provided technology at the pathway level, a gene set enrichment analysis (Gene Set Enrichment Analysis, GSEA) was used to identify the Hallmark gene set (FDR < 0.05) significantly enriched in cells treated by the provided technology (e.g., I-66). The dox-induced CTNNB1-KD and shRNA-resistant CTNNB1 cDNA (shR-cDNA) were included as comparison for rescuing cell lines. GSEA identified the Hallmark Wnt/β -CATENIN gene set, including AXIN2, DKK4, NDK1 and other canonical Wnt target genes, that were significantly down-regulated at 10uM at 6 hours (fdr=0.001) and significantly down-regulated at all 3 doses (1, 3 and 10 uM) at 24 hours and 48 hours (e.g., wnt_beta_catenin_signaling). MYC-targeted and cell cycle-related gene sets (E2F and G2M) were also significantly down-regulated in treated cells by the provided techniques (e.g., I-66), which were first observed at 24 hours and also found at 48 hours (e.g., myc_target_v1, myc_target_v2, e2f_target, g2m_chekpoint, etc.). Changes in these gene sets were determined by dox-induced CTNNB1-KD and reversed by expression of shR-cDNA, indicating that it is indeed a downstream effect of β -catenin. For those enriched in gene sets by CTNNB1-KD (i.e., clotting, myogenesis, interferon), treatment by the provided techniques (e.g., I-66) showed largely consistent trends at 24 hours and 48 hours. In some embodiments, in some evaluations, certain dose/time point combinations may not reach statistical significance. In some embodiments, the present disclosure provides techniques for modulating the expression level and/or function of one or more nucleic acids (e.g., genes) and/or products encoded thereby in one or more such gene sets and/or pathways. In some embodiments, the present disclosure provides techniques for modulating expression and/or function of such gene sets and/or pathways. In some embodiments, the level is reduced. In some embodiments, the level and/or function of expression can be used as a biomarker as described herein, e.g., for assessing treatment, for monitoring treatment progress, for selecting a patient for treatment or continuing treatment, and the like. In some embodiments, negative enrichment of glycolysis and cholesterol gene sets is observed by genetic perturbation rather than by I-66 treatment. In particular, it was determined in various embodiments that the disruption of the interaction of β -catenin with TCF/LEF transcription factors by the provided techniques targets inhibition of β -catenin signaling.
Example 18. Additional characterization and evaluation of the provided techniques.
As described herein, in accordance with the present disclosure, the provided techniques may be characterized and evaluated using a variety of techniques. Certain techniques and results are described herein as examples. Those skilled in the art will appreciate that these exemplary techniques may be adapted or modified.
Crystallization. In some embodiments, structures, interactions, and the like are characterized and evaluated using X-ray crystallography and structural assays. The following schemes are provided as examples. In some embodiments, the β -catenin (human Armadillo) repeat domain 1 to 12 (aa 146 to aa 665))/I-66 complex is concentrated to 9.9mg/mL and a drop tray (drop tray) is set to 4 ℃. In some embodiments, with 0.49M (NH 4 ) 2 SO 4 、0.38M Li 2 SO 4 、0.10M Na 3 Cit, ph=6.00 at 4 ℃. The crystals were then freeze-protected by flash freezing in liquid nitrogen. The 100K diffraction dataset was collected at beam lines PXII and X10SA of the SLS. The molecular replacement solution was obtained using PHASER. In some embodiments, the complete model is built by an iterative loop using both structural refinement of REFMAC and phix, and manual model building in COOT. In some embodiments, the atomic coordinates and the structural factors are stored in a protein database. This structure in particular determines the interaction of various amino acid residues in I-66 with various amino acid residues in β -catenin, for example: PL3-1 and Val349, asp2 and Lys312 and Gly307, npg and Tyr306, asp5 and Asn387 and Trp383,3COOHF-6 and Lys345, ala8 and Trp383, phe9 and Lys345 and Trp383,3Thi-12 and Trp-383 and Asn-415, and BztA-13 and Gln-379, leu-382, val-416, asn-415 and Trp-383 interact.
Competitive fluorescence polarization. In some embodiments, competitive fluorescence polarization is used to assess interactions. The following schemes are described as examples. In some embodiments, 10mM compound in DMSO is serially diluted at 1:3 for a total of 11 concentrations in DMSO using Mosquito LV (SPT Labtech, covina, calif.), then diluted 1000-fold in duplicate in buffer (50mM HEPES,pH 7.5, 125mM NaCl,2% glycerol, 0.5mM EDTA,0.05%v/v pluronic acid) by Mosquito LV, into black polystyrene 384 well plates (Corning, corning, N.Y.). The probe solution was prepared by mixing 10nM full-length beta-catenin (Uniprot ID P35222) with 10nM fluorescently labeled (5 FAM) peptide representing residues 10-53 of TCF4 (Uniprot ID Q9NQB 0) peptide. Plates were incubated at room temperature for 1 hour in the dark prior to reading. The reading was performed on a CLARIOstar reader (BMG Labtech, cary, NC) with excitation at 485nm, emission at 525nm and cut-off at 504 nm. Data were fit to a 1:1 binding model with hill slope using internal script.
SPR. In some embodiments, SPR may be used to characterize or evaluate interactions, binding, etc. The following schemes are described as examples. In some embodiments, at 25 ℃ at Biacore TM SPR experiments were performed on an 8K (Cytiva, marlborough, mass.) instrument. The compounds were diluted into running buffer (50mM Tris pH 8.0, 300mM NaCl,2% glycerol, 0.5mM TCEP,0.5mM EDTA,0.005% tween-20, 1% DMSO). The compounds were diluted to 1uM or 10uM (e.g., peptides A, I-66, etc.) and serially diluted 1:3 to 9 concentrations and two blanks. The series S (cytova) was used to fix biotinylated β -catenin residues 134 to 665 (Uniprot ID P35222) to the active surface of the sensor chip at 10 mL/min for 25 seconds, and compounds were injected to the reference and active surface at 65 mL/min for 180 seconds, then allowed to dissociate for 400 seconds. Using Biacore TM The results were analyzed by the weight evaluation software, double referenced and adapted to the 1:1 binding affinity model.
ABA competition assay. In some embodiments, the provided technology is characterized or assessed using an ABA competition assay. The following schemes are described as examples. In some embodiments, at 25 ℃ at Biacore TM SPR experiments were performed on an S200 (Cytiva) instrument. The β -catenin binding region of APC, E-cadherin and AXIN1, ica t was expressed and purified from E coli (E coli). In some embodiments, BCL9 used is a synthetic peptide comprising an amino acid sequence that interacts with β -catenin. In some embodiments, APCs are treated with a kinase to produce phosphorylated-APCs (papcs) as reported. In some embodiments, the peptide sequence is obtained from the protein database (Protein Data Bank, PDB) or Uniprot: TCF1 (uniprot#P36402, aa 15 to 60), TCF3 (PDB: 1G 3J), TCF4 (PDB: 1 JDH), LEF1 (uniprot#Q UJU2, aa 14 to 62), pAPC (PDB: 1TH 1), mouse E-cadherin (PDB: 1I 7X), human E-cadherin (uniprot#12830, aa 732 to 882), ICAT (PDB: 1 LUJ), AXIN1 (PDB: 1QZ 7), BCL9 (PDB: 2GL 7). The β -catenin binding partner (protein or peptide) was diluted into running buffer (50mM Tris pH 8.0, 300mM NaCl,2% glycerol, 0.5mM TCEP,0.5mM EDTA,0.005% tween-20,0.09% DMSO). Biotinylated β -catenin residues 134 to 665 (uniprot#idp 35222) were immobilized at 10 mL/min to the active surface of the sensor chip for 25 seconds using the biotin CAPture kit, series S (cytova), at a level of about 200RU. The compounds (e.g., I-66, I-470 (as controls), etc.) were diluted to 500nM in running buffer and injected at 90 mL/min for 30 seconds on the surface. In some embodiments, the appropriate concentration of each β -catenin binding partner is selected to ensure > 90% fraction occupancy of the compound (e.g., I-66), and they are injected at 90 uL/min on the surface with increasing or decreasing compound (e.g., I-66) for 67 seconds using the SPR ABA injection protocol. In some embodiments, biacore is used TM The weight evaluation software double referenced and analyzed the results to evaluate competition.
Cell lines and cell cultures. As will be appreciated by those skilled in the art, cell lines are available from a variety of sources, including commercial suppliers. For example, the HAP1 isogenic pair (HZGHC 001062c 011) is available from Horizon Discovery (waters, uk), and many cell lines are available from the american type culture collection (American Type Culture Collection, ATCC). In accordance with the present disclosure, cells may be cultured using a variety of techniques. Cells are routinely cultured in their preferred medium according to the supplier's recommendations. In some embodiments, cells containing the inducible shRNA construct are maintained in a suitable medium with tetracycline-free fetal bovine serum (631101,Clontech Laboratories). A variety of reagents are available from commercial sources. For example, CHIR99021 can be purchased from R & D System (# 4423). In various embodiments, the experiments are typically performed under 4% FBS conditions. In some embodiments, experiments performed at other FBS concentrations are indicated.
NanoBRET. In some embodiments, nanoBRET is used to characterize or evaluate the provided technology. The following schemes are described as examples. In some embodiments, bioluminescence resonance energy transfer (bioluminescence resonance energy transfer, BRET) -based assays were established in HEK293 cells, and β -catenin/TCF 4 interactions (Promega, madison, WI) were assessed using the NanoBRET construct according to the manufacturer's protocol. Fusion of TCF4 to luminescence donor nanoLuc TM And fusing beta-catenin to the receptorNanoBRET TM 618 ligand (HL). Briefly, cells were transfected with NanoBRET plasmid according to the manufacturer's protocol and 30mM (LiCl (L7026, sigma) was added to the cell culture medium to stabilize β -catenin on day 1 fresh medium containing compounds and LiCl was added to the cells on day 2. Nanobretase substrate (N157B, promega) was added to the cells on day 3 and fluorescence emission from HL was measured using GloMAX instrument (Promega) with emissions at 460nM (donor) and 618nM (receiver) cell titer-Glo (CTG) (G7570, promega) was used to monitor cell viability of these cells while NanoBRET analysis.
TCF reporter and negative reporter assay: in some embodiments, the provided technology is characterized or evaluated using a TCF reporter assay. TCF reporter assays including kits have been reported, andmay be used in accordance with the present disclosure. In some embodiments, in a TCF reporter assay, a reporter cell line is generated by reporting lentivirus (79787,BPS Bioscience) using TCF/LEF luciferase and a negative control reporter line is generated using a control luciferase lentivirus (79578,BPS Bioscience). Parental DLD1 cells were transfected with lentivirus and subsequently subjected to 3 days puromycin selection. Monoclonal was selected for both reporter assays. The compounds are incubated with the reporter cells for a suitable period of time, for example 24 hours. The luciferase activity was then measured using the Bright-Glo luciferase assay system (E2620, promega). Cell viability was monitored using a luminescence-based cell viability assay CTG (G7570, promega). Both peptides a and I-66 inhibited luciferase activity in a dose-dependent manner (IC 50 1.5. Mu.M and 0.7. Mu.M, respectively) without affecting cell viability. In the negative control reporter assay, neither showed any activity, with the luciferase under the control of the minimal TATA promoter.
Western blot. A variety of techniques can be used to detect or quantify polypeptides. In some embodiments, western blotting is utilized. The following schemes are described as examples. Cells were harvested in 1 XRIPA buffer (BP-115,Boston Bioproducts) containing a mixture of phosphatase and protease inhibitors (5872S,Cell Signaling Technologies). Tumors were homogenized in 4% SDS buffer using a polytron homogenizer (P000062-PEVO 0-A, bertin). Equivalent amounts of protein were separated on a pre-prepared 4% to 20% SDS-PAGE gel (5671093, bio-Rad) and subsequently transferred to nitrocellulose membrane for detection. In some embodiments, the primary antibody is probed overnight at 4 ℃, and the membrane is washed with TBST and incubated with the appropriate secondary antibody for 1 hour. The membranes were then washed with TBST and visualized using Odyssey imaging system (LI-COR). In some embodiments, the primary antibodies used are β -catenin (8480,Cell Signaling Technology), anti-focal adhesion protein mouse antibody (V9131, sigma-Aldrich), anti-cyclin D2 (3741,Cell Signaling Technology), anti-p 27 (3686,Cell Signaling Technology), anti-HDAC 2 (5113,Cell Signaling Technology). Other antibodies may be used depending on the polypeptide to be evaluated. In some embodiments, the secondary antibodies used are Alexa Fluor 680 secondary antibody (a 32734, thermo Fisher Scientific) and anti-mouse Alexa Fluor 800 secondary antibody (a 32730, thermo Fisher Scientific). In some embodiments, protein bands are visualized and quantified using an Odyssey CLx imaging system (Li-Cor) and ImageStudio software (Li-Cor).
RT-qPCR. In some embodiments, RT-qPCR is used to evaluate transcripts or RNA. The following schemes are described as examples. In some embodiments, the tumor is homogenized in RLT buffer, followed by isolation of total RNA using RNAeasy kit (74104, qiagen) according to the manufacturer's protocol. Cells were washed with ice-cold PBS and total RNA was extracted with RNeasy kit (74104, qiagen). cDNA conversion was performed immediately after RNA extraction using a High capacity cDNA reverse transcription kit (High-Capacity cDNA Reverse Transcription Kit) (4394966, thermoFisher). The cDNA was stored at-20℃until use. qPCR was performed with technical replication on Quantum studio 7Flex real-time PCR system (ThermoFisher) using TaqMan Universal PCR Master Mix (ThermoFisher) and TaqMan Probes (ThermoFisher). The following relative gene expression levels were monitored using Taqman gene expression probes: AXIN2 (Hs 00610344_m1, thermoFisher), SP5 (Hs 01370227_mH, thermoFisher), RNF43 (Hs 00214886_m1, thermoFisher), NOTIUM (Hs 00991061_m1, thermoFisher), CXCL12 (Hs 03676656_mH, thermoFisher). The reactions were performed using Advanced Fast Master Mix (4444557, thermo fisher) and CT values were normalized to ACTB (4325788,Thermo fisher) as an endogenous control. Other suitable probes may be utilized in accordance with the present disclosure.
Co-immunoprecipitation. In some embodiments, co-immunoprecipitation is used to assess interactions, complexation, and the like. The following schemes are described as examples. In some embodiments, peptide A and I-66, but not I-470, dose-dependently block β -catenin/TCF 4 interactions in cells (e.g., DLD1 cells), as detected by Western blotting. In some embodiments, the provided peptides cross cell membranes and/or inhibit β -catenin/TCF interactions. In some embodiments, the provided peptides bind directly to intracellular β -catenin. In some embodiments, it is observed that the plurality of peptides (e.g., I-66) do not affect the β -catenin/E-cadherin interaction, such as in DLD1 cells. In some embodiments, DLD1 cells are treated with the compound for a period of time, e.g., 4 hours, for co-IP experiments. The cell pellet was washed twice with PBS and resuspended in IP-MS cell lysis buffer supplied by Pierce MS-compatible magnetic IP kit (Pierce MS-Compatible Magnetic IP Kit) (90409, thermoFisher (containing a Halt protease/phosphatase inhibitor (78440, thermoFisher)) and sonicated for 2X 10 seconds (30% amplitude) followed by incubation on ice for 10 minutes to effect cell lysis, then the lysate was centrifuged for 10 minutes at 14000 Xg to pellet debris, the final protein concentration was adjusted to about 1mg/mL using lysis buffer (Pierce MS-3725, thermoFisher), 1mL of lysate was added to 96-deep well plates and incubated with rabbit monoclonal β -catenin antibody (8480,Cell Signaling Technology) 1:50 dilution or rabbit isotype control in a thermal mixer at 300rpm for 16 hours at 4℃using a Kifiex magnetic particle processor (Flex 35) and beads (Pierce 35B-35) were eluted at 5X 5 bead 35B-88, beads (Mega) were then eluted at 5X 5 bead 35M, 5 bead 35B-35 bead 35 (Mega) and bead 35 (Mega) was added to the magnetic buffer solution was eluted at 90X 5 bead 35, and the bead 35 bead solution was eluted at 90 bead 35 bead solution (Pierce 35X 35 bead solution) for each condition, thermo fisher) was dried and resuspended in 50 μl of Preomics LYSE buffer and digested according to Preomics iST 96 x kit (P.O.00027, preOimics).
shRNA. In some embodiments, shRNA is used for gene knockdown. In some embodiments, the shRNA construct is prepared in a pLKO-Tet-On lentiviral vector backbone. In some embodiments, the specific sequence targeted is: shNT:5'-CAACAAGATGAAGAGCACCAA-3'; sh637:5'-CTATCAAGATGATGCAGAACT-3'; and sh1787:5'-TCTAACCTCACTTGCAATAAT-3'. The cDNA construct directing the overexpression of CTNNB1 was prepared in a pLVX-EF1 a-IRES-novel lentiviral vector derived from the pLVXEF1a-IRES-puro vector (Clontech, 631988) by replacing the selection cassette. The cDNA construct is not labeled. All constructs were determined by sequencing.
Lentiviral technology. In some embodiments, lentiviral-based constructs (e.g., reporter, shRNA, cDNA overexpression, etc.) are performed using standard protocols from RNAi Consortium (TRC), e.g., from the read Institute (http:// ports. Broadinstitute. Org/gpp/public/resources/protocols). In some embodiments, the shRNA virus is titrated on a single target cell line and infected at an MOI of no greater than 0.7. In some embodiments, the cDNA over-expressed virus is infected, where possible, at a higher MOI. For infection, cells were centrifuged at 2,250rpm for 1 hour in the presence of virus and 8. Mu.g/mL polybrene (H9268, sigma). The medium is pre-placed after rotation and after 24 hours a drug selection (e.g., puromycin or neomycin, as the case may be) is added. Selection is typically performed until all uninfected control cells die.
2D colony formation. In some embodiments, 2D colony formation is used to assess cell growth or proliferation. In some embodiments, COLO320DM cells are plated into 6-well tissue culture plates at 6000 cells/well. The following day, the cells received fresh medium with or without 200ng/mL doxycycline (dox, S5159, selleck). The medium with or without dox was changed every 3 days until the dox-free cells reached 50% to 70% confluence. Cells were fixed with glyoxal (411, ANATECH) at 4℃for 24 hours and then stained with 0.5% crystal violet (031-04852, WAKO) for 1 hour at room temperature. Additional stain was removed with multiple water washes and then imaged by Odyssey CLx imaging system (Li-Cor) and ImageStudio software (Li-Cor).
Proliferation assay. In some embodiments, the provided technology is characterized or assessed using a variety of proliferation assays. In some embodiments, on day 0, the cells are seeded at a desired density (typically 1000 cells/well) in cell culture medium in 96-well plates. On day 1, a 10mM stock solution of compound was first serially diluted into DMSO, followed by dilution with cell culture medium at twice the final concentration. Finally, the culture medium containing the compound is introduced into a cell culture well that already has the same volume of cell culture medium. Cells were incubated with the compounds for the desired days and then lysed against CTG according to the manufacturer's instructions (G7570, promega). Luminescence signals were obtained from a microplate reader (GloMax, promega).
Cell cycle analysis. The effect on the cell cycle by the techniques provided in accordance with the present disclosure can be assessed using a variety of techniques. For example, in some embodiments, COLO320DM cells are prepared for cell cycle analysis using the following protocol: click-iT EdU kit (Thermo Fisher C10337) for monitoring cell proliferation and FxCycle Violet (Thermo Fisher R37166) for DNA quantification. In some embodiments, the flow analysis is performed on a BD LSRFortessa flow cytometer. In some embodiments, compensation is performed between FITC and BV421 channels. In some embodiments, DNA undergoing active synthesis incorporates an EdU dye and is visualized in the FITC channel. In some embodiments, the DNA content incorporates FxCycle dye and is visualized in BV421 channels. Cells were gated into three different populations: low FITC and low BV421 signals (G1 population), high FITC (S population), low FITC and high BV421 (G2 population). Data analysis was performed using FlowJo software (BD Life Sciences).
RNAseq preparation. In some embodiments, RNAseq is used to assess expression of a plurality of nucleic acids including genes. The following describes the procedure as an example. In some embodiments, library preparation and sequencing reactions are performed at GENEWIZ, LLC (South Plainfield, NJ) for RNA-seq of COLO320DM cell line treated with the compound. An RNA-seq library was prepared using the Illumina TruSeqstranded total RNA protocol followed by poly a enrichment. With Illumina HiSeq, an average of 2500 thousands of 2 x 150 base pair reads were generated per sample. For RNA-seq of shRNA treated samples, library preparation and sequencing was performed by Mingma Technologies (Shanghai, china) with a TruSeq chain mRNA library kit on the Novaseq S4 platform with poly a enrichment. On average, each sample produced over 6000 ten thousand 2×150 base pair reads. For RNA-seq of cell line transplanted tumors, library preparation and sequencing was performed by Fulgent Gentetics (Houston, TX) with the Truseq chain mRNA library kit on the Novaseq S4 platform with PolyA enrichment.
RNAseq data analysis. In accordance with the present disclosure, RNAseq data can be analyzed using a variety of techniques. In some embodiments, trimmings of sequence are used to remove possible aptamer sequences and poor quality nucleotides. The trimmed reads were mapped to the homo sapiens GRCh38 reference gene set available on ENSEMBL using STAR aligner v.2.7.7a. For transplanted tumor samples, host reads were removed with XenofilteR. Unique gene hit counts were calculated by using the featureCount from the R sub packet v.2.4.2. Readout filtration, normalization and differential expression analysis were performed with the edge package v.4.0.2 in R. In some embodiments, for each comparison, the gene with an adjusted p-value < 0.01 and absolute log2 fold change > 1 is referred to as a differentially expressed gene. Genes differentially expressed in at least one comparison were used for heat map and cluster analysis. In some embodiments, gene expression is normalized to fold change compared to a simultaneous DMSO control. In some embodiments, the R pheeatmap package v.1.0.12 is used to make a heat map and for hierarchical clustering of genes with relevance as a similarity measure.
In some embodiments, for enrichment analysis, GSEA v4.1.0 is run with the MSigDB database v7.3 by fold change arranged gene list. A Venn plot was generated with ggvenn v.0.1.9, where overlapping p-values were calculated using the hypergeometric test in rv4.1.2.
And extracting nucleoprotein. In some embodiments, the nucleoprotein is extracted for evaluation. The following schemes are described as examples. Using a supplement with Hall TM NE-PER of protease and phosphatase inhibitor cocktail (78442,Thermo Fisher Scientific) TM Nuclear and cytoplasmic extraction reagents (78833,Thermo Fisher Scientific) cytoplasmic and nuclear proteins were performed according to manufacturer's protocolAnd (5) extracting quality. The cytoplasmic and nuclear extracts were stored at-80 ℃ until use.
Immunofluorescent staining. In some embodiments, immunofluorescent staining is used to detect, quantify, characterize, or evaluate the polypeptide. The following schemes are described as examples. In some embodiments, COLO320DM cells are seeded at an initial density of 40,000 cells/chamber in RPMI and 10% FBS with Nunc TM Lab-Tek TM IIChamber Slide TM Overnight in system (154534PK,Thermo Fisher Scientific). The next day, the medium was replaced with RPMI with 4% FBS containing 0.1% DMSO, 10uM I-66 or I-470. After 24 hours of compound treatment, cells were washed with PBS and fixed with 10% neutral buffered formalin (HT 501128-4L, sigma-Aldrich) for 15 minutes at room temperature. Cells were then simultaneously permeabilized and blocked with 0.1% Triton X-100 (X100-100 ML, sigma-Aldrich) and 10% donkey serum (D9663-10 ML, sigma-Aldrich) in PBS for 1 hour at room temperature. Thereafter, cells were incubated overnight at 4℃with anti- β -catenin rabbit primary antibody (8480,Cell Signaling Technology) diluted 1:100 (v/v) in 0.1% Triton x-100/10% donkey serum/PBS permeation/blocking buffer. Cells were then incubated simultaneously with anti-rabbit Alexa Fluor 488 secondary antibody (a 32790, thermo Fisher Scientific) diluted 1:1000 (v/v) and phalloidin Alexa Fluor 647 (a 30107, thermo Fisher Scientific) diluted 1:200 (v/v) in 0.1% Triton x-100/10% donkey serum/PBS permeation/blocking buffer for 1 hour at room temperature. Those skilled in the art will appreciate that other primary and/or secondary antibodies may also be used. Thereafter, the cells were then incubated with DAPI (D3571, thermo Fisher Scientific) diluted 1:10000 in PBS for 15 minutes at room temperature. After each step the cells were washed with PBS for 3 x 5 min. The chamber walls were then removed and the ProLong was used at room temperature TM Glass anti-blocking agent (P36980, thermo Fisher Scientific) cells were blocked overnight with cover glass. Cells were imaged using a Zeiss LSM 710 confocal laser scanning system. Confocal images were analyzed using FIJI/ImageJ.
Animal studies. In some embodiments, the provided techniques are evaluated using animal models. Experiments are generally in institutionsThe animal care and use committee approved protocol and followed institutional guidelines for proper and humane use of animals. The following schemes are described as examples. For example, for COLO320DM xenograft assessment, male NU/J mice (6 to 8 weeks old) were used and when the average tumor volume reached 300mm 3 Mice were randomized at this time. For IP administration, the compounds were formulated as 10mg/mL arginine and 6% PEG400 phosphate (pH 7.4) formulations. In some embodiments, the PDX murine model is established in athymic nude-Foxn 1 nu female mice, e.g., in some embodiments, CRC patient tumors with APC mutations (Tyr 935 Ter), amplified HER2, wild-type KRAS and β -catenin, and high AXIN2 expression therein. Tumor volume was measured by electronic calipers every 2 to 3 days until tumor volume reached 2000mm 3 And is estimated as (length×width 2 )/2. Weigh every 2 to 3 days. Tumor Growth Inhibition (TGI) was calculated as: TGI% = [1 (TVi-TV 0)/(Tvi-TVv 0)]X 100% (TVi: average tumor volume of the group administered at day i, TV0: average tumor volume of the group administered at day 0, TVI: average tumor volume of the group vehicle at day i, TVv0: average tumor volume of the group vehicle at day 0). On the last day specified, by CO 2 Animals were euthanized by asphyxiation and plasma, tumors, tissues, etc. were removed for further analysis.
Quantification of the compounds. In some embodiments, LC-MS is used to quantify a variety of compounds, including stapled peptides. In some embodiments, the concentration of a compound (e.g., a stapled peptide) in a biological sample is measured by LC-MS/MS (triple quadrupole 6500+). Using analytical grade chemicals and solvents, 25ng/mL tolbutamide in acetonitrile (ACN, LS120-4,Fisher Scientific) was used as an internal standard. 8. Mu.L of plasma or tissue lysates were used in LC method, mobile phase A (H 2 1% formic acid in O (FA, LS118-4,Fisher Scientific)), mobile phase B (0.1% FA in ACN), flow 0.6 ml/min, column at Waters ACQUITY UPLC BEH C, 2.1 x 50mm,1.7 um. Calibration curves were generated using 5 to 5000ng/mL stapled peptide (e.g., I-66, I-470, etc.) in mouse plasma and tissue homogenates. In some embodiments, ionization and multiple reactions are achieved by electrospray The monitoring scan proceeds to the MS. In some embodiments, PK parameters (e.g., plasma maximum concentration (Cmax) and AUC) are analyzed by the non-compartmental model 200 of Phoenix WinNonlin 8.3.3 using a linear/logarithmic trapezoidal method.
Plasma NOTUM by mass spectrometry. The following protocol is described as one example of plasma NOTUM by mass spectrometry. In some embodiments, plasma samples are collected from mice transplanted with COLO320DM tumors for bird gun proteome analysis. In some embodiments, the highest abundance of proteins, e.g., the first 3, in a plasma sample is first removed using a multiple affinity removal column (Multiple Affinity Removal Column) Mouse-3 (4.6x50 mm,5188-4217, agilent) (immunoaffinity HPLC-based method). In some embodiments, the removal of highly abundant proteins allows for medium to low abundance proteins to be detected by the bird gun method proteomics. UltiMate TM A 3000 quick disconnect quaternary system (Rapid Separation Quaternary System) (thermo fisher) is configured as suggested in the operating guidelines. For each sample 45uL was added to 180uL of Agilent buffer A (5185-5987) and centrifuged at 16,000Xg for 1 min in a 0.22um spin filter (5185-5990, agilent). 180uL of each sample was injected onto a Mouse-3 column. The low/high abundance proteins eluted from the Mouse-3 column were monitored by UV detector at 280 nm. The low abundance proteins were collected by fraction collectors. The final volume of each low abundance fraction is about 1mL. Each fraction was concentrated at 3,400 Xg for 60 minutes using a 5kDa MWCO spin column concentrator (5185-8991, agilent). After this step is completed, the sample volume is about 50 to 80uL. Samples were digested with trypsin (25200114, thermo fisher) using the PreOmics iST 96 x digestion kit (p.o.00027) protocol.
For LC-MS/MS analysis of peptide mixtures, separation was performed using the UltiMate 3000RSLCnano system (thermo fisher). Peptides were isolated based on hydrophobicity using the following: EASY-Spray PepMap RSLC C, 2um,500mm x 75 μm i.d. column, under constant temperature control at 50 ℃ and flow rate of 300 nL/min, and linear gradientThe total duration was 90 minutes for from 2% to 30% acetonitrile (0.1% FA). After the gradient portion of the chromatogram, the column was washed with 99% acetonitrile containing 0.1% FA for 14 minutes and equilibrated with 2% acetonitrile containing 0.1% FA for 26 minutes. In some embodiments, the MS analysis was performed on Q exact HF-X (Thermo Fisher) in positive ion mode using an EASY-spray source (ES 903, thermoFisher). The instrument was operated at a spray voltage of 1.9kV, an ion transfer capillary temperature of 250 ℃ and an S-lens RF level of 40%. A 120,000 resolution high resolution FTMS scan comprising 1 microscanning with a maximum injection time of 200 MS followed by 18 15,000 resolution dependent FTMS MS/MS scans with a maximum injection time of 28 MS. The dependent MS/MS scan was performed using a separation width of 1.4m/z of the target electrophile. The separated multiple charged ions (2, 3, 4) are activated using HCD normalized collision energy of 28 eV. To prevent ions from triggering a subsequent data dependent scan after the data dependent scan has been triggered, a 30 second dynamic exclusion is enabled.
In some embodiments, the proteome discoverer v2.5.0.400 is used for protein identification and quantification using the sequence HT algorithm. For plasma proteomics experiments, database searches were performed using the Homo sapiens (sp_clinical taxid=9606) (v 2021-07-30) and the mouse (Mus musculus) database (sp_clinical taxid=10090) (v 2021-09-30). Database searches were performed with the following settings: trypsin digestion, 20ppm precursor mass tolerance, 0.02Da fragment mass tolerance, static modification: ureidomethyl, dynamic modification: oxidation/N-terminal Met-loss. Protein abundance in each sample was normalized to total protein mass and normalized protein abundance of NOTUM was extracted. Comparison of average normalized NOTUM abundance between groups was performed by one-way ANOVA followed by Tukey HSD. For the co-immunoprecipitation experiments, database searches were performed using the homo sapiens database and the same settings as the plasma proteomes described above. Average normalized abundance of β -catenin binding partners was compared between conditions by one-way ANOVA followed by Tukey HSD. In some embodiments, the mass spectrometry proteomics data is deposited, for example, by PRIDE partner reservoirs to ProteomeXchange alliance (http:// Proteomexchange. Org).
Example 19 a compound comprising a thioether staple like structure may provide a variety of activities.
As described herein, a variety of staple-like structures may be used in accordance with the present disclosure. In some embodiments, the staple-like structure comprises-S-. In some embodiments, the staple-like structure comprises two-S-. In some embodiments, the two-S-are not bonded to each other. In some embodiments, the staple-like structure is a thioether staple-like structure. A variety of such staple-like structures are described herein, e.g., having-L s1 -S-L s2 -S-L s3 -a staple-like structure of structure, wherein L s1 、L s2 And L s3 Each independently as described herein. In some embodiments, L s1 And L s3 Independently from amino acid residues such as cysteine, homocysteine, alpha-methyl cysteine, penicillamine, and the like. In some embodiments, each is-CH 2 -. In some embodiments, two thiol groups are formed by reacting a thiol group with a thiol having LG-L s2 -compounds of the LG structure or salts thereof are linked by reaction, wherein LG and L s2 Each independently as described herein. A variety of such compounds are as described herein. In some embodiments, such staple-like structures are (i, i+4) staple-like structures. In some embodiments, such staple-like structures are closer to the C-terminus. In some embodiments, such staple-like structure is at X 10 And X is 14 Between them. The present disclosure particularly determines that stapled peptides comprising such staple-like structures can provide a variety of activities, e.g., binding to a target (e.g., β -catenin), inhibiting tumor growth, etc. Some of the stapled peptides and data are shown below as examples. C-1 is Ac-PL3-Asp-Npg-B5-Asp-3COOHF-Ala-Ala-Phe-Leu-pyrS2-2F3MeF-BztA-Gln-NH2, wherein PL3 and B5, and B5 and pyrS2 are stapled. In some embodiments, C-1 is the second generating peak/fraction on HPLC of the formulation Ac-PL3-Asp-Npg-B5-Asp-3COOHF-Ala-Ala-Phe-Leu-pyrS2-2F3MeF-BztA-Gln-NH2 (see, e.g., table E2).
It was determined that various stapled peptides inhibited cell proliferation. For example, in an assay to evaluate COLO320 viability, the IC50 s of I-1271, I-1274, I-1278 and C-1 showed IC50 s of 900nM, 3.4uM, 2.4uM and 4.1uM, respectively.
As determined herein, certain amino acid residues (e.g., cys/Cys) and/or staple-like structures (e.g., as in I-1271, I-1272, I-1274, I-1275, etc.) provide stronger binding and activity (e.g., inhibit cell proliferation) than other stapled peptides.
Although various embodiments have been described and illustrated herein, a variety of other means and/or structures for performing the functions and/or obtaining the results and/or one or more advantages described in this disclosure will be readily apparent to those of ordinary skill in the art, and each of such variations and/or modifications is deemed to be included. More generally, those skilled in the art will readily appreciate that all parameters, dimensions, materials, and configurations described herein are meant to be exemplary and that the actual parameters, dimensions, materials, and/or configurations will depend upon the specific application or applications for which the teachings of the present disclosure are used. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the disclosure described in the present disclosure. It is, therefore, to be understood that the foregoing embodiments are presented by way of example only, and that the provided techniques (including those claimed) may be practiced otherwise than as specifically described and claimed. In addition, if two or more features, systems, articles, substances, kits, and/or methods are not mutually inconsistent, any combination of two or more such features, systems, articles, substances, kits, and/or methods is included within the scope of the present disclosure.

Claims (87)

1. An agent having the following structure or a salt thereof:
2. an agent, wherein the agent is or comprises:
[X] p X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 [X] p’ ，
wherein:
p15, p16 and p17 are each independently 0 or 1;
p and p' are each independently 0 to 10; and is also provided with
X，X 1 ，X 2 ，X 3 ，X 4 ，X 5 ,X 6 ,X 7 ，X 8 ，X 9 ，X 10 ，X 11 ，X 12 ，X 13 ，X 14 ，X 15 ，X 16 And X 17 Each independently is an amino acid residue.
3. The agent of claim 4, wherein the agent is or comprises a peptide comprising:
[X 0 ] p0 X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 ，
wherein:
p0, p15, p16 and p17 are each independently 0 or 1;
X 0 ，X 1 ，X 2 ，X 3 ，X 4 ，X 5 ，X 6 ，X 7 ，X 8 ，X 9 ，X 10 ，X 11 ，X 12 ，X 13 ，X 14 ，X 15 ，X 16 and X 17 Each independently is an amino acid residue, wherein:
X 2 comprising a side chain containing an acidic or polar group;
X 5 comprising a side chain containing an acidic or polar group; and is also provided with
X 9 、X 12 And X 13 Each comprising a side chain containing an optionally substituted aromatic group.
4. The agent of any one of the preceding claims comprising three or more staple-like structures within 10 to 20 amino acid residues.
5. The agent of any one of the preceding claims wherein X 0 ，X 1 ，X 3 ，X 4 ，X 7 ，X 10 ，X 11 And X 14 Each independently is an amino acid residue suitable for stapling, or each independently is stapling.
6. The agent of any one of claims 1 to 5, wherein X 1 And X 4 Connected by a staple-like structure.
7. The agent of any one of claims 1 to 5, wherein X 0 And X 4 Connected by a staple-like structure.
8. The agent of any one of claims 1 to 7, wherein X 4 And X 11 Connected by a staple-like structure.
9. The agent of any one of claims 1 to 8, wherein X 10 And X 14 Connected by a staple-like structure.
10. The agent of any one of claims 1 to 9, wherein X 7 And X 10 Connected by a staple-like structure.
11. The agent of any one of claims 1 to 10, wherein X 7 And X 14 Connected by a staple-like structure.
12. The agent of any one of the preceding claims, wherein the agent comprises an N-terminal group.
13. The agent of any one of the preceding claims wherein X 1 Is a residue of an amino acid having the structure of formula A-I, A-II or A-III, wherein R a1 And R is a3 Taken together with intervening atoms, form an optionally substituted 3-to 10-membered ring having 0 to 5 heteroatoms in addition to the intervening atoms.
14. The agent of any one of the preceding claims wherein X 1 Is PL3.
15. Any of the preceding claimsThe medicament, wherein X 4 Is the residue of an amino acid comprising an alkene.
16. The agent of any one of the preceding claims wherein X 4 Is B5.
17. The agent of any one of the preceding claims wherein X 10 Is a residue of an amino acid comprising an optionally substituted carboxy, an optionally substituted amino, an azide, an optionally substituted alkynyl or an optionally substituted thiol.
18. The agent of any one of the preceding claims wherein X 10 Is Lys.
19. The agent of any one of the preceding claims wherein X 11 Is the residue of an amino acid comprising an alkene.
20. The agent of any one of the preceding claims wherein X 11 Is pyrS2.
21. The agent of any one of the preceding claims wherein X 14 Is the residue of an amino acid comprising a carboxyl, amino, azido, alkynyl or thiol group.
22. The agent of any one of the preceding claims wherein X 14 Is GlnR.
23. The agent of any one of the preceding claims wherein X 10 And X 14 One of which is a residue of an amino acid comprising a carboxyl group and the other is a residue of an amino acid comprising an amino group.
24. The agent of any one of the preceding claims wherein X 10 And X 14 Connected by a staple-like structure, wherein the staple-like structure comprises-C (O) N (R') -.
25. The agent of any one of the preceding claims wherein X 2 Comprising a side chain containing an acidic group.
26. The agent of any one of claims 1 to 24 wherein X 2 Is that
Asp, ala, asn, glu, npg, ser, hse, val, S5, S6, aeLys, tfeGA, aThr, aad, pro, thr, phe, leu, PL3, gln, isoGlu, meAsn, isoDAsp, rbGlu, sbGlu, aspSH, ile, sbMeAsp, rbMeAsp, aMeDAsp, OAsp,3COOHF,NAsp,3Thi,NGlu,isoDGlu,BztA,Tle,Aib,MePro,Chg,Cha, or DipA.
27. The agent of any one of the preceding claims wherein X 3 Comprising one or two hydrophobic side chains.
28. The agent of any one of claims 1 to 26 wherein X 3 Npg, ile, asp, cha, dipA, chg, leu, B5, cba, S5,
ala, glu, allylGly, nLeu, ser, B6, asn, B4, glnR, val, [ Phc ] [ allyl ] Dap, hse, [ Bn ] [ allyl ] Dap,1MeK, R5, phe, cypA, cyLeu, pff, diethA, tyr, trp, aib, phg, octG, morphNva, F2PipNva, [ Piv ] [ allyl ] Dap, [ CyCO ] [ allyl ] Dap, lys, or S3.
29. The agent of any one of the preceding claims wherein X 5 Comprising a side chain containing an acidic group.
30. The agent of any one of claims 1 to 28 wherein X 5 Selected from Asp,3COOHF, tfeGA, gln,
[ CH2CMe2CO2H ] TriAzDap, thr, glu,2OH3COOHF, 4COOHF,2COOHF, his, tyr,5F3Me2COOHF,4F3Me2COOHF,5F3Me3COOHF,4F3Me3COOHF,3F2COOHF, val, ser, trp, asn, ala, arg, dGlu, aThr, hTyr,3cbmf, leu, phe, lys, and Ile.
31. The agent of any one of the preceding claims wherein X 6 Comprising a side chain containing an acidic group.
32. The agent of any one of the preceding claims wherein X 6 Is 3COOHF, tfEGA or Asp.
33. The agent of any one of the preceding claims wherein X 7 Is a hydrophobic amino acid residue.
34. The agent of any one of claims 1 to 32 wherein X 7 Selected from Aib, ala, morphGLn, gln, ser,
iprllys, nLeu, cha, hse, npg, val, cyLeu, thr, phe, acp, asn, daMeS, aMeDF, leu, cpg, cbg, me2 gin, met2O, acLys, his, aMeL, daMeL, aMeV, aMeS, and aMeF.
35. The agent of any one of the preceding claims wherein X 8 Is a hydrophobic amino acid residue.
36. The agent of any one of claims 1 to 34 wherein X 8 Selected from Ala, aib, cpg, val, leu, gln,
lys, asp, glu, aad, nLeu, cba, ser, thr, agr, morph, phe, hPhe, hTyr, and AcLys.
37. The agent of any one of the preceding claims wherein X 9 Comprising such side chains: the side chain is or comprises an optionally substituted aromatic group.
38. The agent of any one of claims 1 to 36 wherein X 9 AA9, phe, ala, lys,3COOHF, aib,2NapA,
nLeu,2Thi, tyr,3Thi,4FF,4ClF,4BrF,3FF,3C1F,3BrF,2FF,3OMeF,4CNF,3CNF,4MeF,3MeF, aic, rbipPrF, sbipPrF, rbipPrDF, rbMeXyleA, rbMeXyleDA, cba, cypA, bztA, INapA, trp, leu, ile, ser, chg, hse,4TriA,3F3MeF, thr, his, val, asn, gln,2Cpg, sbMeXyleA, or SbMeXyleDA.
39. The agent of any one of the preceding claims wherein X 12 Comprising such side chains: the side chain is or comprises an optionally substituted aromatic group.
40. The agent of any one of claims 1 to 38 wherein X 12 Is that
3Thi, phe,2F3MeF, pyrS2,2ClF, hnLeu, bztA,2Thi,2MeF,2FF,34ClF, lys, nLeu,2COOHF,2PhF, hCBA, hCypA, hpe, dipA, hepG, dap7Abu, hhLeu, hhSer, hexG, [2IAPAc ]2NH2F, ala, abu, leu, hleu, npg, cpa, pyrS1, [ Bnc ]2NH2F, [ Phc ]2NH2F, [ Biph ]2NH2F, [3PyAc]2NH2F,Nva,Cba,ChA,2FurA,2OMeF,2BrF,2CNF,2NO2F,2PyrA,3PyrA,4PyrA,His,1NapA,Val,Ile,Chg,DiethA,OctG,2cbm F,c6Phe, [ MePipAc ]2NH2F, or [2PyCypCO ]2NH2F.
41. The agent of any one of the preceding claims wherein X 13 Comprises optionally substituted aromatic groups.
42. The agent of any one of claims 1 to 40 wherein X 13 Selected from BztA, 34ClF, 2NapA, 3BrF, 34MeF, 3Thi, phe, glnR, 34MeF, 2NapA and Lys.
43. The agent of any one of the preceding claims, wherein p15 is 1.
44. The agent of any one of the preceding claims wherein X 15 Comprising hydrophobic side chains.
45. The agent of any one of the preceding claims, wherein the peptide forms a structure comprising a helix.
46. The agent of any one of the preceding claims, wherein the peptide binds to β -catenin.
47. The agent of any one of the preceding claims, wherein the peptide and sequence is seq id NO:2 or a sequence comprising SEQ ID NO:2 or a fragment thereof:
SVLFYAITTLHNLLLHQEGAKMAVRLAGGLQKMVALLNKTNVKFLAITTDCLQILAYGNQESKLIILASGGPQALVNIMRTYTYEKLLWTTSRVLKVLSVCS SNKPAIVEAGGMQALGLHLTDPSQRLVQNCLWTLRNLSDAATKQEGMEGLLGTLVQLLGSDDINVVTCAAGILSNLTCNNYKNKMMVCQVGGIEALVRT(SEQ ID NO：2)。
48. the agent of any one of the preceding claims, wherein the peptide binds to β -catenin and interacts with one or more residues that are or correspond to: SEQ ID NO:1, or at least three, or at least four, or at least five, or at least six, or at least seven, or at least eight, or at least nine, or at least ten, or at least eleven, or at least twelve, or at least thirteen, or at least fourteen, or at least fifteen, or at least sixteen, or at least seventeen, or at least eighteen, or at least nineteen, or at least twenty of the following amino acid residues at the positions indicated in 1: a305 Y306, G307, N308, Q309, K312, R342, K345, V346, V349, Q375, R376, Q379, N380, L382, W383, R386, N387.D413, N415, V416, T418, and C419.
49. The medicament of any of the preceding claims, wherein the double bond of the (i, i+7) staple-like structure is E.
50. The medicament of any of the preceding claims, wherein the double bond of the (i, i+7) staple-like structure is Z.
51. The medicament of any of the preceding claims, wherein the double bond of the (i, i+2), (i, i+3) or (i, i+4) staple-like structure is E.
52. The medicament of any of the preceding claims, wherein the double bond of the (i, i+2), (i, i+3) or (i, i+4) staple-like structure is Z.
53. The agent of any one of the preceding claims, wherein the carbon atom bonded to both staple-like structures (e.g., in B5) is in the R configuration.
54. The agent of any one of the preceding claims, wherein the carbon atom bonded to both staple-like structures (e.g., in B5) is in the S configuration.
55. An agent having the following structure or a salt thereof:
SP-1-1，SP-1-2，SP-1-3，SP-1-4，SP-I-5，SP-1-6，SP-1-7，SP-1-8，SP-2-1，SP-2-2，SP-2-3,SP-2-4，SP-2-5，SP-2-6，SP-2-7，SP-2-8，SP-3-1，SP-3-2，SP-4-1，SP-4-2，SP-4-3，SP-4-4，SP-4-5，SP-4-6，SP-4-7，SP-4-8，SP-5-1，SP-5-2，SP-5-3，SP-5-4，SP-5-5，SP-5-6，SP-5-7，SP-5-8，SP-6，SP-7-1，SP-7-2,SP-7-3，SP-7-4,SP-7-5,SP-7-6,SP-7-7，SP-7-8，SP-8-1，SP-8-2，SP-8-3，SP-8-4，SP-8-5，SP-8-6，SP-8-7，SP-8-8，SP-9-1，SP-9-2，SP-9-3，SP-9-4，SP-9-5，SP-9-6，SP-9-7，SP-9-8，SP-10-1，SP-10-2，SP-10-3，SP-10-4，SP-10-5，SP-10-6，sP-10-7，SP-10-8，SP-11-1，SP-11-2，SP-11-3，SP-11-4，SP-11-5，SP-11-6，SP-11-7，SP-11-8，SP-12-1，SP-12-2，SP-12-3，SP-12-4,SP-12-5，SP-12-6,SP-12-7,SP-12-8，SP-13-1，SP-13-2,SP-13-3,SP-13-4,SP-13-5，SP-13-6，SP-13-7，SP-13-8，SP-14-1，SP-14-2，SP-14-3，SP-14-4，SP-14-5，SP-14-6，SP-14-7，SP-14-8，SP-15-1，SP-1 5-2，SP-15-3，SP-15-4，SP-15-5，SP-15-6，SP-15-7，SP-15-8。
56. an agent having the following structure or a salt thereof:
57. an agent having the following structure or a salt thereof:
58. the agent of any one of claims 56 to 57, wherein the agent has the same retention time as I-66 prepared as described in example 9 under HPLC conditions that separate I-66 and I-67 prepared as described in example 9.
59. The agent of any one of claims 56 to 57, wherein the agent exhibits a retention time of about 15.3 minutes under the following HPLC conditions: agilent Poroshell 120EC-C18; 4.6X100 mm; solvent a = 0.1% tfa in water; solvent B = 0.075% tfa in acetonitrile; gradient 10% b to 95% b over 30 minutes; the UV absorbance at 220nM is detected.
60. The agent of any one of claims 56 to 59, wherein the agent elutes in a single peak with I-66 prepared as described in example 9 under the following HPLC conditions: agilent Poroshell 120EC-C18; 4.6X100 mm; solvent a = 0.1% tfa in water; solvent B = 0.075% tfa in acetonitrile; gradient 10% b to 95% b over 30 minutes; the UV absorbance at 220nM is detected.
61. The agent of any one of claims 56 to 60, wherein said agent exhibits an overlap with a peak between about 5.1 to 5.7 in figure 6 under the same or equivalent conditions 1 H NMR peaks.
62. The pharmaceutical formulation of any one of claims 56 to 60, which exhibits a concentration of between about 5.1 and about 5.7 identical to figure 6 under identical or equivalent conditions 1 H NMR peaks.
63. The pharmaceutical formulation of any one of claims 56 to 60, wherein under identical or equivalent conditions, at which 1 Corresponding to bonding to carbon atoms in the H NMR spectrum 1 The peak of H overlaps with the peak in fig. 6.
64. The pharmaceutical formulation of any one of claims 56 to 60, which is under identical or equivalent conditions 1 The H NMR spectrum overlaps with the peaks in fig. 6.
65. The agent of any one of claims 56 to 57, wherein the agent has the same retention time as I-67 prepared as described in example 9 under HPLC conditions that separate I-66 and I-67 prepared as described in example 9.
66. The agent of any one of claims 56 to 57, wherein the agent exhibits a retention time of about 16.2 minutes under the following HPLC conditions: agilent Poroshell 120EC-C18; 4.6X100 mm; solvent a = 0.1% tfa in water; solvent B = 0.075% tfa in acetonitrile; gradient 10% b to 95% b over 30 minutes; the UV absorbance at 220nM is detected.
67. The agent of any one of claims 56 to 59, wherein the agent elutes in a single peak with I-67 prepared as described in example 9 under the following HPLC conditions: agilent Poroshell 120EC-C18; 4.6X100 mm; solvent a = 0.1% tfa in water; solvent B = 0.075% tfa in acetonitrile; gradient 10% b to 95% b over 30 minutes; the UV absorbance at 220nM is detected.
68. The agent of any one of claims 56 to 59 and 65 to 67, wherein said agent is shown to be the same orUnder comparable conditions, do not overlap with peaks between about 5.1 and 5.7 in FIG. 6 1 H NMR peaks.
69. The agent of any one of claims 56 to 59 and 65 to 67, wherein said agent does not exhibit a concentration of between about 5.1 and 5.7 that is the same as figure 6 under the same or equivalent conditions 1 H NMR peaks.
70. The pharmaceutical formulation of any one of claims 56 to 59 and 65 to 67, wherein under identical or equivalent conditions, under which 1 Corresponding to bonding to carbon atoms in the H NMR spectrum 1 The peaks of H do not all overlap with the peaks in fig. 6.
71. The pharmaceutical formulation of any one of claims 56 to 60, which is under identical or equivalent conditions 1 The H NMR spectrum does not overlap with the peaks in fig. 6.
72. The agent of any one of the preceding claims, wherein the carbon atom bonded to both staple-like structures (e.g., in B5) is in the S configuration.
73. An agent having the structure of formula I:
R N -L P1 -L AA1 -L P2 -L AA2 -L P3 -L AA3 -L P4 -L AA4 -L PS -L AaS -L P6 -L AA6 -L P7 -R C ，
I
wherein:
R N is a peptide, an amino protecting group or R' -L RN -；
L P1 、L P2 、L P3 、L P4 、L P5 、L P6 And L P7 Each independently is L, wherein L P1 、L P2 、L P3 、L P4 、L P5 、L P6 And L P7 Comprising:
a first R' groupAnd a second R' group taken together to form-L s -, the-L s -bonding to an atom to which a first R 'group is attached and an atom to which a second R' group is attached; and
a third R 'group and a fourth R' group taken together to form-L s -, the-L s -bonding to an atom to which a third R 'group is attached and an atom to which a fourth R' group is attached;
each L s Independently is-L s1 -L s2 -L s3 -, each L s1 、L s2 And L s3 Independently is L;
L AA1 is an amino acid residue comprising a side chain containing an acidic or polar group;
L AA2 is an amino acid residue comprising a side chain containing an acidic or polar group;
L AA3 is an amino acid residue;
L AA4 is an amino acid residue comprising a side chain comprising an optionally substituted aromatic group;
L AA5 is an amino acid residue comprising a side chain comprising an optionally substituted aromatic group;
L AA6 is an amino acid residue comprising a side chain comprising an optionally substituted aromatic group;
R C is a peptide, a carboxyl protecting group, -L RC -R’、-O-L RC -R 'or-N (R') -L RC -R’；
L RN And L RC Each independently is L;
each L is independently a covalent bond or an optionally substituted divalent C 1 -C 25 Aliphatic or heteroaliphatic groups having from 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -，Cy-，-O-，-S-，-S-S-，-N(R’)-，C(O)-，C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 N (R') -C (O) S-, or-C (O) O-;
each-Cy-is independently an optionally substituted divalent 3-to 30-membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms;
each R' is independently-L-R, -C (O) R, -CO 2 R or-SO 2 R；
Each R is independently-H or an optionally substituted group selected from: c (C) 1-30 Aliphatic, C having 1 to 10 heteroatoms 1-30 Heteroaliphatic, C 6-30 Aryl, C 6-30 Arylaliphatic, C having 1 to 10 heteroatoms 6-30 Aryl heteroaliphatics, 5 to 30 membered heteroaryl groups having 1 to 10 heteroatoms and 3 to 30 membered heterocyclyl groups having 1 to 10 heteroatoms, or
The two R groups optionally and independently together form a covalent bond, or:
two or more R groups on the same atom optionally and independently taken together with the atom form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the atom; or alternatively
Two or more R groups on two or more atoms optionally and independently taken together with their intervening atoms form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the intervening atoms; or alternatively
An agent having the structure of formula I:
R N -L P1 -L A^1 -L P2 -L AA2 -L P3 -L AA3 -L P4 -L AA4 -L P5 -L AA5 -L P6 -L AA6 -L P7 -R C ，
I
wherein:
R N is a peptide, an amino protecting group or R' -L RN -；
L P1 、L P2 、L P3 、L P4 、L P5 、L P6 And L P7 Each independently is L, wherein L P1 、L P2 、L P3 、L P4 、L P5 、L P6 And L P7 Comprising:
a first R 'group and a second R' group taken together to form-L s -, the-L s -bonding to an atom to which a first R 'group is attached and an atom to which a second R' group is attached; and
a third R 'group and a fourth R' group taken together to form-L s -, the-L s -bonding to an atom to which a third R 'group is attached and an atom to which a fourth R' group is attached;
each L s Independently is-L s1 -L s2 -L s3 -, each L s1 、L s2 And L s3 Independently is L;
L AA1 is L AR Wherein the methylene units are represented by-C (R') (R AS ) -substitution, wherein R AS is-L AS1 -R AA1 Wherein R is AA1 is-CO 2 R or-SO 2 R；
L AA2 Is L AR Wherein the methylene units are represented by-C (R') (R AS ) -substitution, wherein R AS is-L AS2 -R AA2 Wherein R is AA2 is-CO 2 R or-SO 2 R；
L AA3 Is L AR Wherein the methylene units are represented by-C (R') (R AS ) -substitution, wherein R AS is-L AS3 -R AA3 Wherein R is AA3 Is R';
L AA4 is L AR Wherein the methylene units are represented by-C (R') (R AS ) -substitution, wherein R AS is-L AS4 -R AA4 Wherein R is AA4 Is an optionally substituted group selected from: a 6 to 14 membered aryl or a 5 to 14 membered heteroaryl having 1 to 6 heteroatoms;
L AA5 is L AR Wherein the methylene units are represented by-C (R') (R AS ) -substitution, wherein R AS is-L AS5 -R AA5 Wherein R is AA5 Is an optionally substituted group selected from: a 6 to 14 membered aryl or a 5 to 14 membered heteroaryl having 1 to 6 heteroatoms;
L AA6 is L AR Wherein the methylene units are represented by-C (R') (R AS ) -substitution, wherein R AS is-L AS6 -R AA6 Wherein R is AA6 Is an optionally substituted group selected from: a 6 to 14 membered aryl or a 5 to 14 membered heteroaryl having 1 to 6 heteroatoms;
R C is a peptide, a carboxyl protecting group, -L RC -R’、-O-L RC -R 'or-N (R') -L RC -R’；
L RN And L RC Each independently is L;
each L AR Independently an optionally substituted divalent C 1 -C 6 An aliphatic group wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -、-C(R’)(R AS )-，-Cy-，-O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 N (R') -C (O) S-, or-C (O) O-;
L AS1 、L AS2 、L AS3 、L AS4 、L AS5 and L AS6 Each independently is L AS ；
Each R AS Independently is-L AS -R’；
Each L AS Independently a covalent bond or an optionally substituted divalent C 1 -C 10 Aliphatic or heteroaliphatic groups having from 1 to 5 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -，-Cy-，O-，-S-，-S-S-，-N(R’)-，-C(O)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O) 2 -，-S(O) 2 N (R') -C (O) S-, or-C (O) O-;
each L is independently a covalent bond or an optionally substituted divalent C 1 -C 25 Aliphatic or heteroaliphatic groups having 1 to 10 heteroatoms, wherein one or more methylene units of the group are any Optionally and independently replaced by: -C (R') 2 -Cy-, -O-, -S-, -N (R '), -C (O) -, -C (S) -, -C (NR '), -C (O) N (R '), -N (R ') C (O) O-, -S (O) 2N (R '), -C (O) S-, or-C (O) O-;
each-Cy-is independently an optionally substituted divalent 3-to 30-membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms;
each R' is independently-L-R, -C (O) R, -CO 2 R or-SO 2 R；
Each R is independently-H or an optionally substituted group selected from: c (C) 1-30 Aliphatic, C having 1 to 10 heteroatoms 1-30 Heteroaliphatic, C 6-30 Aryl, C 6-30 Arylaliphatic, C having 1 to 10 heteroatoms 6-30 Aryl heteroaliphatics, 5 to 30 membered heteroaryl groups having 1 to 10 heteroatoms and 3 to 30 membered heterocyclyl groups having 1 to 10 heteroatoms, or
The two R groups optionally and independently together form a covalent bond, or:
two or more R groups on the same atom optionally and independently taken together with the atom form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the atom; or alternatively
Two or more R groups on two or more atoms optionally and independently together with their intervening atoms form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the intervening atoms.
74. The medicament of any one of the preceding claims, wherein each olefinic double bond in the staple-like structure is independently and optionally converted to a single bond.
75. The agent of any one of the preceding claims having a diastereomeric purity of about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more, or a purity of about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more.
76. A pharmaceutical composition comprising or delivering the agent or amino acid of any one of the preceding claims, and a pharmaceutically acceptable carrier.
77. A composition selected from table E2 or table E3; or a pharmaceutical composition comprising or delivering one or more or all of the peptide agents selected from the compositions of table E2 or table E3, and a pharmaceutically acceptable carrier.
78. The composition of any one of the preceding claims, comprising a medicament comprising one or more staple-like structures, each independently comprising one or more olefinic double bonds, wherein the ratio of the two stereoisomers of olefinic double bonds in the staple-like structures is about 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 30:1, 40:1, 50:1 or higher.
79. A method, comprising:
a) Preparing a first compound comprising two moieties, each independently comprising an olefinic double bond;
b) Stapling the two parts to form a first formed staple-like structure by olefin metathesis between the olefin double bond in one part and the olefin double bond in the other part, thereby providing a second compound;
c) Adding one or more additional moieties to the second compound to provide a third compound comprising two moieties, each independently comprising an olefinic double bond; and
d) The two portions of the third compound are stapled to form a second formed staple-like structure by olefin metathesis between an olefin double bond in one portion and an olefin double bond in the other portion, providing a fourth compound.
80. A method for modulating β -catenin interaction with a partner in a system comprising contacting β -catenin with the agent or composition of any of the preceding claims; or alternatively
A method for modulating β -catenin interaction with a partner in a system comprising administering or delivering to the system the agent or composition of any of the preceding claims; or alternatively
A method for modulating TCF- β -catenin interactions in a system comprising contacting β -catenin with the agent or composition of any of the preceding claims; or alternatively
A method for modulating TCF- β -catenin interactions in a system comprising administering or delivering to the system the agent or composition of any of the preceding claims; or alternatively
A method for inhibiting β -catenin dependent cell proliferation comprising administering or delivering to the system the agent or composition of any of the preceding claims; or alternatively
A method for modulating the WNT/β -catenin pathway in a system, comprising administering or delivering to the system the agent or composition of any one of the preceding claims, wherein expression of a nucleic acid is modulated; or alternatively
A method comprising administering or delivering the agent or composition of any one of the preceding claims to the system, wherein the level of nucleic acid transcripts and/or products thereof is modulated; or alternatively
A method comprising administering or delivering the agent or composition of any one of the preceding claims to the system, wherein expression of the nucleic acid is modulated.
81. A method for treating or preventing a condition, disorder or disease associated with β -catenin and a partner interaction in a subject, comprising administering or delivering to the subject an effective amount of the agent or composition of any of the preceding claims, preferably wherein the partner is TCF7, LEF1, TCF7L2, axin1, axin2 or APC.
82. A method for treating cancer in a subject comprising administering or delivering to the subject an effective amount of the agent or composition of any one of the preceding claims.
83. The method of any one of the preceding claims, comprising administering or delivering a second therapeutic agent or treatment to the subject.
84. The method of any one of the preceding claims, wherein the second therapeutic agent is or comprises: a chemotherapeutic agent, a hormonal therapeutic agent, an immunotherapeutic agent, a checkpoint inhibitor, an antibody, CTLA-4, PD-1 or PD-L1 inhibitor, or a cell; or the second treatment is or comprises the following: surgery, chemotherapy, radiation therapy, hormone therapy, stem cell or bone marrow transplantation, immunotherapy, T cell therapy, or CAR T cell therapy.
85. The method of any one of the preceding claims, comprising assessing expression of the nucleic acid.
86. A compound having the formula PA structure or a salt thereof:
N(R PA )(R a1 )-L a1 -C(R a2 )(R a3 )-L a2 -C(O)R PC ，
PA
wherein:
R PA is-H or an amino protecting group;
R a1 and R is a3 Each independently is-L a -R’；
R a2 is-L aa -C(O)R PS ；
L a 、L a1 And L a2 Each independently is L;
-C(O)R PS -COOH, optionally protected or activated;
-C(O)R PC -COOH, optionally protected or activated;
each L is independently a covalent bond or an optionally substituted divalent C 1 -C 25 Aliphatic or heteroaliphatic groups having from 1 to 10 heteroatoms, wherein one or more methylene units of the group are optionally and independently replaced by: -C (R') 2 -，-Cy-，-O-，-S-，-S-S-，-N(R’)-，-C(o)-，-C(S)-，-C(NR’)-，-C(O)N(R’)-，-N(R’)C(O)N(R’)-，-N(R’)C(O)O-，-S(O)-，-S(O)2-，-S(O) 2 N (R') -C (O) S-, or-C (O) O-;
each-Cy-is independently an optionally substituted divalent 3-to 30-membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms;
each R' is independently-R, -C (O) R, -CO 2 R or-SO 2 R is R; and is also provided with
Each R is independently-H or an optionally substituted group selected from: c (C) 1-30 Aliphatic, C having 1 to 10 heteroatoms 1-30 Heteroaliphatic, C 6-30 Aryl, C 6-30 Arylaliphatic, C having 1 to 10 heteroatoms 6-30 Aryl heteroaliphatics, 5 to 30 membered heteroaryl groups having 1 to 10 heteroatoms and 3 to 30 membered heterocyclyl groups having 1 to 10 heteroatoms, or
The two R groups optionally and independently together form a covalent bond, or:
two or more R groups on the same atom optionally and independently taken together with the atom form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the atom; or alternatively
Two or more R groups on two or more atoms optionally and independently taken together with their intervening atoms form an optionally substituted 3 to 30 membered monocyclic, bicyclic or polycyclic ring having 0 to 10 heteroatoms in addition to the intervening atoms; or alternatively
A compound having the following structure or a salt thereof:
wherein:
R PA is-H or an amino protecting group;
-C(O)R PS -COOH, optionally protected or activated; and is also provided with
-C(O)R PC -COOH, optionally protected or activated; or alternatively
A compound having the following structure or a salt thereof:
wherein:
R PA is-H or an amino protecting group;
-C(O)R PS -COOH, optionally protected or activated; and is also provided with
-C(O)R PC Is an optionally protected or activated-COOH.
87. The agent, compound, method or composition of any one of embodiments 1 to 2401.
CN202280053893.9A 2021-06-08 2022-06-08 Stapled peptides and methods thereof Pending CN117794560A (en)

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