JPWO2021028686A5

JPWO2021028686A5 -

Info

Publication number: JPWO2021028686A5
Application number: JP2022508564A
Authority: JP
Publication date: 2023-08-25

Description

(多量体結合複合体)
一実施態様において、修飾された多量体結合複合体は、以下の表1に記載されている結合複合体を含む:
表1:例示されている本発明の修飾された多量体結合複合体

(multimeric binding complex)
In one embodiment, the modified multimeric binding complex comprises a binding complex set forth in Table 1 below:
Table 1: Illustrative Modified Multimeric Binding Complexes of the Invention

¹¹C、¹⁸F、¹⁵O、及び¹³Nなどの陽電子放出同位体による置換は、標的占有率を調べるための陽電子放出トモグラフィー(PET)試験において有用であり得る。 Substitution with positron emitting isotopes, such as ¹¹ C, ¹⁸ F, ¹⁵ O, and ¹³ N, can be useful in positron emission tomography (PET) studies for examining target occupancy.

(BCY15985)

化合物1(30.0mg、6.02μmol、1.0当量)、化合物2(8.49mg、9.03μmol、1.5当量)の混合物をDMF(0.5mL)に溶解させた。この溶液のpHを0.1M DIEA(7.7mg、60.17μmol、10.5μL、10.0当量)の滴加により8に調整した。反応混合物を25℃で1.0時間撹拌した。LC-MSにより、BCY15459が完全に消費され、所望のm/zを有する1つの主要なピーク(計算されたMW: 5811.8、観測されたm/z: 1454.3([M/4+H]⁺)、1163.4([M/5+H]⁺)、969.7([M/6+H]⁺)、831.4([M/7+H]⁺))が検出され、BCY15459－2当量のビオチン-PEG12-NHS(計算されたMW: 6637.8、観測されたm/z: 1328.7([M/5+H]⁺))も観察されることが示された。反応混合物を濾過し、減圧下で濃縮すると、残渣が得られた。粗生成物を分取HPLC(TFA条件)により精製すると、BCY15985(21mg、3.53μmol、58.61％収率、97.6％純度)が白色の固形物として得られた。 (BCY15985)

A mixture of compound 1 (30.0 mg, 6.02 μmol, 1.0 eq) and compound 2 (8.49 mg, 9.03 μmol, 1.5 eq) was dissolved in DMF (0.5 mL). The pH of this solution was adjusted to 8 by dropwise addition of 0.1 M DIEA (7.7 mg, 60.17 μmol, 10.5 μL, 10.0 equiv). The reaction mixture was stirred at 25° C. for 1.0 hour. LC-MS showed complete consumption of BCY15459 and one major peak with the desired m/z (MW calculated: 5811.8, m/z observed: 1454.3 ([M/4+H] ⁺ ) , 1163.4 ([M/5+H] ⁺ ), 969.7 ([M/6+H] ⁺ ), 831.4 ([M/7+H] ⁺ )) were detected, and BCY15459-2 equivalents of biotin -PEG12 -NHS (MW calculated: 6637.8, m/z observed: 1328.7 ([M/5+H] ⁺ )) was also shown to be observed. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The crude product was purified by preparative HPLC (TFA conditions) to give BCY15985 (21 mg, 3.53 μmol, 58.61% yield, 97.6% purity) as a white solid.

Claims

A multimeric binding complex comprising at least two bicyclic peptide ligands,
The peptide ligands may be the same or different, each of which is a polypeptide comprising at least three cysteine residues separated by at least two loop sequences and said cysteine residues of said polypeptide. a molecular scaffold that forms a covalent bond such that at least two polypeptide loops are formed on the molecular scaffold;
The multimeric binding complex further comprising a modifier group conjugated thereto, wherein the modifier group comprises a tracer molecule, a detectable moiety, or a lipid.

2. The multimeric binding complex of claim 1, wherein said tracer molecule is a fluorophore selected from fluorescein, Alexa Fluor(TM) 488, cyanine-5, and BODIPY(TM) FL.

Multimer binding according to claim 1, wherein the detectable moiety is a binding detectable moiety, such as a biotin-containing moiety, in particular a biotin-containing and pegylated moiety, such as biotin-Peg4 and biotin-Peg12. Complex.

2. The multimeric binding complex of claim 1, wherein said lipid is a palmitoyl-containing moiety.

A multimeric binding complex according to any one of claims 1 to 4, comprising a compound of formula (I):

(Where CHM represents the central hinge portion;
S ₁ and S ₂ represent spacer groups;
bicyclic ₁ and bicyclic ₂ represent the bicyclic peptide ligands of claim 1;
m represents an integer selected from 1 to 9; and modifier represents a modifier group according to any one of claims 1 to 4).

A multimeric binding complex according to any one of claims 1 to 4, comprising a compound of formula (II):

(Where CHM represents the central hinge portion;
S ₁ and S ₂ represent spacer groups;
bicyclic ₁ represents a bicyclic peptide ligand according to claim 1;
n represents an integer selected from 2 to 10; and modifier represents a modifier group according to any one of claims 1 to 4).

A multimeric binding complex according to claim 5 or claim 6, wherein m and n represent integers selected from 2-9, eg 2 or 3.

8. A multimeric binding complex according to claim 7, wherein m and n represent 3 and CHM is the motif of formula (A):

(Wherein, “-----” represents the point of attachment to each spacer group (S ₁ or S ₂ )).

8. A multimeric binding complex according to claim 7, wherein n represents 3 and CHM is the motif of formula (B):

(Wherein, "-----" represents the point of attachment to the spacer group; and

represents the point of attachment to the modifier group).

8. A multimeric binding complex according to claim 7, wherein n represents 2 and CHM is a motif of formula (C):

(Wherein, "-----" represents the point of attachment to the spacer group; and

represents the point of attachment to the modifier group).

8. A multimeric binding complex according to claim 7, wherein n represents 2 and CHM is a motif of formula (D):

(Wherein, "-----" represents the point of attachment to the spacer group; and

represents the point of attachment to the modifier group).

Claims 5-, wherein said spacers (S ₁ and S ₂ ) are selected from any one of spacers S _A , S _B , S _C , S _D , S _E , S _F , S _G and S _H 12. The multimeric binding complex of any one of 11:

(Wherein, "-----" represents the point of attachment to the CHM group; and

represents the point of attachment to said bicyclic or modifier group, eg, S _A , where n is 5, 10, or 23, eg, 10 or 23).

A multimeric binding complex according to any one of claims 5 to 11, wherein said spacers (S ₁ and S ₂ ) are absent.

14. The multimeric binding complex of any one of claims 1-13, wherein at least one of said peptide ligands is specific for CD137, eg each of said peptide ligands is specific for CD137.

15. The multimeric binding complex of any one of claims 1-14, wherein said loop sequence comprises 6 amino acids.

wherein the peptide ligand is

(where C _i , C _ii , and C _iii represent the first, second, and third cysteine residues, respectively, Nle represents norleucine, PYA represents propargylic acid, and tBuAla represents t -butyl-alanine)
16. The multimeric binding complex of any one of claims 1-15, comprising a core amino acid sequence selected from: or a pharmaceutically acceptable salt thereof.

said peptide ligand comprises N- and C-terminal additions, and
Ac-A-(SEQ ID NO: 1)-[Dap(PYA)]- _CONH2 (hereinafter referred to as BCY7741);
Ac-A-(SEQ ID NO: 1)-[Dap(Lys(PYA))]- _CONH2 (hereinafter referred to as BCY12799);
Ac-(SEQ ID NO: ₂ )-A-Pra-CONH2 (hereinafter referred to as BCY7077);
Ac-A-(SEQ ID NO:3)-A- _CONH2 (hereinafter referred to as BCY7744);
Ac-A-(SEQ ID NO:3)-K- _CONH2 (hereinafter referred to as BCY11613);
Ac-[dA]-(SEQ ID NO: 4)-[dA] _-CONH2 (hereinafter referred to as BCY11506);
Ac-[dA]-(SEQ ID NO:5)-[dK] _-CONH2 (hereinafter referred to as BCY12144);
Ac-(SEQ ID NO:6)-A- _CONH2 (hereinafter referred to as BCY8927);
Ac-(SEQ ID NO:6)-K- _CONH2 (hereinafter referred to as BCY12357);
Ac-(SEQ ID NO:7)-A (referred to herein as BCY8928); and
Ac-(SEQ ID NO:7)-K (referred to herein as BCY13389)
(where Dap represents diaminopropionic acid, PYA represents propargylic acid, and Pra represents propargylglycine).
17. The multimeric binding complex of claim 16, comprising an amino acid sequence selected from: or a pharmaceutically acceptable salt thereof.

Claims 1-, wherein said molecular scaffold is 1,1',1''-(1,3,5-triazinane-1,3,5-triyl)triprop-2-en-1-one (TATA) 18. The multimeric binding complex according to any one of 17.

19. The multimeric binding complex of any one of claims 1-18 listed in Table 1, with the exception of the unbound control BCY12374.

at least one (e.g., two) of said peptide ligands is specific for CD137 and at least one (e.g., one) of said peptide ligands is nectin-4, e.g. 14. The multimeric binding complex of any one of claims 1-13, which is specific for the listed.

At least one (e.g., two) of said peptide ligands is specific for CD137 and at least one (e.g., one) of said peptide ligands is EphA2, e.g., listed in Table 3 14. The multimeric binding complex of any one of claims 1-13, which is specific for a

22. The multimeric binding complex of any one of claims 1-21, wherein said pharmaceutically acceptable salt is selected from the free acid or sodium, potassium, calcium, ammonium salts.

23. The multimeric binding complex of any one of claims 12-22, wherein said CD137 is human CD137.

A drug conjugate comprising a multimeric binding complex according to any one of claims 1-23 conjugated to one or more effectors and/or functional groups.

A pharmaceutical composition comprising a multimeric binding complex according to any one of claims 1-23 or a drug conjugate according to claim 24.

A multimeric binding complex according to any one of claims 1 to 23, a drug conjugate according to claim 24, or a claim for use in the prevention, suppression or treatment of a disease or disorder mediated by CD137 Item 26. The pharmaceutical composition according to Item 25.

Use of a multimeric binding complex according to any one of claims 1-23 in an analytical method (ie as tracer or tag).