WO2024027795A1 - Antibody-drug conjugate containing myc protein degradation agent bioactive compound, preparation method therefor, and use thereof - Google Patents

Antibody-drug conjugate containing myc protein degradation agent bioactive compound, preparation method therefor, and use thereof Download PDF

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WO2024027795A1
WO2024027795A1 PCT/CN2023/111003 CN2023111003W WO2024027795A1 WO 2024027795 A1 WO2024027795 A1 WO 2024027795A1 CN 2023111003 W CN2023111003 W CN 2023111003W WO 2024027795 A1 WO2024027795 A1 WO 2024027795A1
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group
alkyl
aryl
antibody
pharmaceutically acceptable
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PCT/CN2023/111003
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French (fr)
Chinese (zh)
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童友之
薛彤彤
宋帅
肖亮
蔡家强
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苏州开拓药业股份有限公司
苏州宜联生物医药有限公司
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Publication of WO2024027795A1 publication Critical patent/WO2024027795A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems

Definitions

  • the present disclosure belongs to the field of medical technology and relates to antibody-drug conjugates of protein-degrading bioactive compounds, protein-degrading bioactive compounds, drug linker conjugates and their preparation methods, as well as their use in prevention and/or treatment.
  • Diseases related to abnormal cell activity including but not limited to use in the prevention and/or treatment of tumor diseases.
  • TPD Targeted Protein Degradation
  • PROTAC protein degradation targeting chimera
  • MMD molecular glue degrader
  • Myc is an important family of transcription factors, including c-Myc, n-Myc, and l-Myc, which are essential for cell growth, metabolism and tissue development. At the same time, Myc is also an important group of oncogenes. c-Myc is abnormally expressed in about 30-50% of human malignant tumors and plays a role in affecting cell growth, proliferation, differentiation, apoptosis, as well as cell metabolism and malignant transformation. plays an extremely important role. Abnormally high expression of c-Myc is associated with poor prognosis in many human cancers.
  • c-Myc due to the special protein structure of c-Myc, it directly targets inhibitors Therapeutic inhibition of c-Myc has been unsuccessful, and approaches targeting the c-Myc transcriptional machinery have had limited success. Therefore, c-Myc has long been considered an undruggable target.
  • targeted protein degraders and molecular glue degraders can change the target from “undruggable” to “druggable”.
  • drug properties such as low oral bioavailability
  • off-target The resulting dose-limiting toxicity and other problems have greatly restricted the development of this type of molecules.
  • the present invention relates to antibody drug conjugates (ADCs) and their uses.
  • ADC drugs combine the tumor targeting effect of antibodies and the high activity of biologically active compounds, becoming a biological missile with very promising efficacy and safety advantages.
  • the solubility is greatly improved, enabling intravenous administration and effectively solving pharmaceutical problems such as the bioavailability of small molecule compounds.
  • Antibodies guide ADCs to bind to target cells, achieving tumor tissue enrichment, reducing non-target tissue exposure, and reducing possible toxicity caused by systemic administration of bioactive compounds.
  • ADCs that bind to tumor cells can cleave and release bioactive molecules in the tumor microenvironment to kill tumor cells.
  • ADC can also be internalized by tumor cells and undergo enzymatic decomposition under the action of specific enzymes in the cells to release small molecule drugs to treat diseases. Therefore, it is expected that an ADC composed of a protein degrading agent and a tumor-targeting antibody can achieve tumor enrichment, eliminate or reduce the toxic side effects caused by the protein degrading agent acting on non-diseased tissues, and improve the therapeutic effect. Using ADC technology to achieve tumor targeting of protein degradation agents and reduce their toxic and side effects will have high clinical value.
  • the present disclosure provides an antibody drug conjugate represented by Formula I,
  • Tb is an antibody or its antigen-binding fragment
  • S is the sulfur atom on Tb
  • L is the linker, which covalently binds Tb and D;
  • D is the Myc protein degrader fragment
  • q is an integer selected from 1-20.
  • L is a linker that covalently binds Tb and D
  • linker L is covalently bound to the S atom on Tb and is covalently bound to D. Similar expressions below have similar meanings.
  • an antibody drug conjugate represented by Formula II is provided,
  • L 1 is a linker unit, which covalently binds the antibody or its antigen-binding fragment (Tb) to L 2 ;
  • L 2 is a connecting unit, which covalently binds the linker unit (L 1 ) and L 3 ;
  • L 3 is selected from amino acid residues or a short peptide consisting of 2-10 (such as 2, 3, 4, 5, 6, 7, 8, 9 or 10) amino acid residues; the amino acid residues are selected from natural Amino acid residues, unnatural amino acid residues or amino acid residues shown in AA 1 or their stereoisomers;
  • R x or R y is hydrogen, and the other is selected from Alternatively, R x and R y and the carbon atoms to which they are commonly connected form a 4-10 membered heterocyclic ring, which is optionally substituted by one or more R 0 ;
  • R x1 and R y1 are each independently selected from hydrogen and C 1-6 alkyl; alternatively, R x1 and R y1 and the nitrogen atoms commonly connected to them form a 4-10 membered heterocyclic ring, and the 4-10 membered heterocycle
  • the heterocycle is optionally substituted with one or more R 0' ;
  • R 0 and R 0' are each independently selected from C 1-6 alkyl, C 3-6 cycloalkyl, -NR x2 R y2 and 4-10 membered heterocyclyl optionally substituted by C 1-6 alkyl. ;
  • R x2 and R y2 are each independently selected from hydrogen and C 1-6 alkyl;
  • L 4 is a chemical bond or spacer unit, covalently combining D and L 3 ;
  • Tb, q and D are as defined in any aspect of this disclosure.
  • the q is selected from 1-18, 1-16, 1-14, 1-12, 1-10, 1-8, 2-8, or 4-6; for example, is selected from 1-1 An integer of 10.
  • said q is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10.
  • said q is selected from 2, 4, 6 and 8.
  • L is selected from Position 1 is connected to Tb through the S atom, and position 2 is connected to L 2 .
  • L is selected from Position 1 is connected to Tb through the S atom, and position 2 is connected to L 2 .
  • L2 is selected from Bit 1 is connected to L 1 , bit 2 is connected to L 3 ,
  • W is selected from -CH 2 -, -OCH 2 CH 2 -,
  • W' is selected from -CR x3 R y3 - and -NR x3 -,
  • R x3 and R y3 are each independently selected from hydrogen, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 3-6 cycloalkyl, and 3-6 membered heterocyclyl,
  • R x3 and R y3 together with the carbon atoms connected to them form a 3-6 membered carbocyclic ring or a 3-6 membered heterocyclic ring,
  • p is any integer selected from 0-10.
  • W is -CH2- .
  • W' is selected from -CH 2 -, -C(CH 3 ) 2 -, -N(CH 3 )- and -NH-.
  • R x3 and R y3 are each independently selected from hydrogen and methyl.
  • p is selected from 0, 1, 2, 3 and 4.
  • L2 is selected from Bit 1 is connected to L 1 , bit 2 is connected to L 3 .
  • L 1 -L 2 are selected from Position 1 is connected to Tb through an S atom, position 2 is connected to L3 , and W, W' and p are defined as described in any scheme of this disclosure.
  • L 1 -L 2 are selected from
  • Position 1 is connected to Tb through the S atom, and position 2 is connected to L 3 .
  • L 1 -L 2 are selected from Position 1 is connected to Tb through the S atom, and position 2 is connected to L 3 .
  • L 3 is selected from the amino acid residues represented by AA 1 or includes 2-10 of the amino acid residues represented by AA 1 (e.g., 2, 3, 4, 5, 6, 7, 8, 9 or 10 ) A short peptide composed of amino acid residues; the amino acid residues are selected from natural amino acid residues, non-natural amino acid residues or amino acid residues represented by AA 1 or their stereoisomers.
  • L3 is selected from Val-Cit, Val-Ala, Phe-Lys, Val-Lys, Val- AA1 , Ala-Ala-Ala, Ala-Ala-Asn, Val- AA1 -Gly, Gly-Gly-Phe-Gly.
  • L3 is selected from Val- AA1 -Gly.
  • the structure of the amino acid residue represented by AA 1 is as follows:
  • R x or R y is hydrogen, and the other is selected from R x1 and R y1 are each independently selected from hydrogen and C1-6 alkyl.
  • R x1 and R y1 are each independently selected from methyl, ethyl, n-propyl and n-butyl; preferably, R x1 and R y1 are both n-propyl.
  • L4 is selected from chemical bonds
  • Bit 1 is connected to L 3 and bit 2 is connected to D.
  • L 4 is Bit 1 is connected to L 3 and bit 2 is connected to D.
  • D is a Myc protein degrading agent fragment, such as a c-Myc protein degrading agent fragment, an n-Myc protein degrading agent fragment, etc.
  • D is a c-Myc protein degrader fragment.
  • D is a structural unit represented by Formula III, and D is connected to L 4 through the oxygen atom, sulfur atom or nitrogen atom it contains;
  • Q is selected from: -NR 2 -, -O-,
  • Ring A represents a heterocycloalkyl group containing at least one N atom as a heteroatom, which is connected to T or R 1 through an N atom. Ring A is optionally substituted by one or more groups selected from R 9 ;
  • T, U, and Z are each independently selected from: chemical bond, N, O, carbonyl group, C 1 -C 6 alkylene group, C 3 -C 10 cycloalkylene group, arylene group, heteroarylene group, heterocyclic ring Base, the alkylene, cycloalkylene, arylene, heteroarylene, heterocyclylene is optionally substituted by one or more R 9 ;
  • Group substitution of C 1 -C 3 alkyl, C 1 - C 3 alkoxy or -OP( O)(OM) 2 ;
  • M is independently selected from: hydrogen, C 1 -C 4 alkyl
  • R 11 and R 12 are each independently selected from: hydrogen, C 1 -C 4 alkyl, aryl, aryl-alkyl;
  • R 21 is selected from: chemical bond, C 1 -C 4 alkylene
  • R 22 and R 23 are each independently selected from: hydrogen, C 1 -C 4 alkyl, aryl, aryl-alkyl, the C 1 -C 4 alkyl, aryl, aryl-alkyl is optional Substituted with a group selected from halogen, hydroxyl, and amino;
  • a is selected from: 0, 1, 2, 3, 4 or 5;
  • b is selected from: 0, 1, 2, 3, 4 or 5;
  • n is selected from: 0, 1, 2 or 3;
  • n is selected from: 0, 1, 2, 3 or 4;
  • the structural unit represented by formula III contains at least one group selected from R 13 or R 14 .
  • D is a structural unit represented by Formula III, and D is connected to L 4 through the oxygen atom or nitrogen atom it contains;
  • Q is selected from: -NR 2 -, -O-,
  • Ring A represents a heterocycloalkyl group containing at least one N atom as a heteroatom, which is connected to T through an N atom. Ring A is optionally substituted by one or more groups selected from R 9 ;
  • T, U, and Z are each independently selected from: chemical bond, carbonyl group, C 1 -C 6 alkylene group, C 3 -C 10 cycloalkylene group, arylene group, heteroarylene group, heterocyclylene group, the above Alkylene, cycloalkylene, arylene, heteroarylene, heterocyclylene are optionally substituted by one or more R 9 ;
  • M is independently selected from: hydrogen, C 1 -C 4 alkyl
  • R 11 and R 12 are each independently selected from: hydrogen, C 1 -C 4 alkyl, aryl, aryl-alkyl;
  • R 21 is selected from: chemical bond, C 1 -C 4 alkylene
  • R 22 and R 23 are each independently selected from: hydrogen, C 1 -C 4 alkyl, aryl, aryl-alkyl, the C 1 -C 4 alkyl, aryl, aryl-alkyl is optional Substituted with a group selected from halogen, hydroxyl, and amino;
  • a is selected from: 0, 1, 2, 3, 4 or 5;
  • b is selected from: 0, 1, 2, 3, 4 or 5;
  • n is selected from: 0, 1, 2 or 3;
  • n is selected from: 0, 1, 2, 3 or 4;
  • the structural unit represented by formula III contains at least one group selected from R 13 or R 14 .
  • the R n is selected from: hydroxyl-substituted C 1 -C 8 alkyl; more preferably, the R n is selected from: hydroxyl substituted C 1 -C 4 alkyl; most preferably, the R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH or -CH 2 CH 2 CH 2 CH 2 OH.
  • R m is selected from: hydrogen, -C 1 -C 4 alkyl.
  • the R n is selected from: hydroxyl, mercapto or amino-substituted C 1 -C 8 alkyl; preferably, the R n is selected from: hydroxyl, mercapto or amino substituted C 1 -C 4 Alkyl group; more preferably, the R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 CH 2 SH.
  • R m is selected from: hydrogen, -C 1 -C 4 alkyl.
  • D is a structural unit represented by Formula III, and D is connected to L 4 through any hydroxyl group in R 13 or R 14 . In some embodiments, D is a structural unit represented by Formula III, and D is connected to L 4 through any hydroxyl, amino or thiol group in R 13 or R 14 .
  • the R 9 is selected from: hydrogen, C 1 -C 4 alkyl, R 13 ; more preferably, the R 9 is selected from: R 13 .
  • the R 13 is selected from: hydroxyl, R n , -OR n .
  • the R 13 is selected from: hydroxyl, thiol, amino, R n , -OR n .
  • the R n is selected from: hydroxyl-substituted C 1 -C 8 alkyl; more preferably, the R n is selected from: hydroxyl-substituted C 1 -C 4 alkyl; most preferably, the R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH or -CH 2 CH 2 CH 2 CH 2 OH.
  • the R n is selected from: hydroxyl, mercapto or amino-substituted C 1 -C 8 alkyl; preferably, the R n is selected from: hydroxyl, mercapto or amino substituted C 1 -C 4 Alkyl group; more preferably, the R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH.
  • the R a is selected from: phenyl, naphthyl, carbazolyl, 1-azacarbazolyl, 2-azacarbazolyl, 1,8-diazacarbazolyl, Indolyl, 7-azaindolyl, 2,3-dihydroindolyl, 2,3-dihydro-7-azaindolyl, phenoxazinyl, fluorenyl, quinolinyl, iso Quinolyl, naphthyridinyl, tetralinyl, tetrahydroquinolinyl, pyrimidinyl, triazolyl, bicyclo[1.1.1]pentyl, norbornyl, adamantyl.
  • the R a is optionally substituted by one or more R 9 , and the R 9 is selected from hydrogen, R 13 , halogen, cyano, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, C 1 -C 6 alkoxy, phenyl, naphthyl.
  • the R a is selected from: phenyl, naphthyl, carbazolyl, 1-azacarbazolyl, 2-azacarbazolyl, 1,8-diazacarbazolyl, Indolyl, 7-azaindolyl, 2,3-dihydroindolyl, 2,3-dihydro-7-azaindolyl, phenoxazinyl, fluorenyl, quinolinyl, iso Quinolyl, naphthyridinyl, tetralinyl, tetrahydroquinolinyl, pyrimidinyl, pyridyl, quinolyl-pyridyl, triazolyl, bicyclo[1.1.1]pentyl, norbornane base, adamantyl.
  • R 13 halogen, cyano, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, C 1 -C 6 alkoxy, phenyl, naphthyl.
  • the R a is selected from: phenyl, naphthyl-1-yl, naphthyl-2-yl, carbazol-9-yl, 1-azacarbazol-9-yl, 2-azacarbazole- 9-yl, 1,8-diazacarbazol-9-yl, indol-1-yl, 2,3-dihydro-indol-1-yl, 7-azaindol-1-yl , 2,3-dihydro-7-azaindolyl-1-yl, phenoxazin-10-yl, fluoren-9-yl, quinolin-4-yl, quinolin-5-yl, quinoline -8-yl, isoquinolin-1-yl, isoquinolin-4-yl, isoquinolin-5-yl, isoquinolin-8-yl, 1,2,3,4-tetrahydro-1, 8-naphthyridin-1-yl, 1,2,3,4
  • the R a is optionally substituted by one or more R 9 , and the R 9 is selected from: R 13 , F, Cl, Br, cyano, methyl, ethyl, trifluoromethyl, methoxy base, ethoxy, phenyl, naphthyl-1-yl, naphthyl-2-yl.
  • the R a is selected from: phenyl, naphth-1-yl, naphth-2-yl, carbazol-9-yl, 1-azacarbazol-9-yl, 2-aza Carbazol-9-yl, 1,8-diazacarbazol-9-yl, indol-1-yl, 2,3-dihydro-indol-1-yl, 7-azaindolyl- 1-yl, 2,3-dihydro-7-azaindolyl-1-yl, phenoxazin-10-yl, fluoren-9-yl, quinolin-4-yl, quinolin-5-yl , quinolin-8-yl, isoquinolin-1-yl, isoquinolin-4-yl, isoquinolin-5-yl, isoquinolin-8-yl, 1,2,3,4-tetrahydro -1,8-naphthyridin-1-yl, 1,2,3,4-tetra
  • R a is optionally substituted by one or more R 9 , and the R 9 is selected from: R 13 , F, Cl, Br, cyano, methyl, ethyl, trifluoromethyl, methoxy, Ethoxy, phenyl, naphthyl-1-yl, naphthyl-2-yl.
  • the R n is selected from: hydroxyl-substituted C 1 -C 8 alkyl; more preferably, the R n is selected from: hydroxyl substituted C 1 -C 4 alkyl; most preferably, R n is selected from From: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, or -CH 2 CH 2 CH 2 CH 2 OH.
  • the R n is selected from: hydroxyl hydroxyl, mercapto or amino-substituted C 1 -C 8 alkyl; preferably, the R n is selected from: hydroxyl, mercapto or amino substituted C 1 -C 4 alkyl; more preferably, the R n is selected from : -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH.
  • the R m is selected from: hydrogen, -C 1 -C 4 alkyl.
  • the R 9 is selected from: F, Cl, Br, cyano, methyl, ethyl, trifluoromethyl, methoxy, ethoxy, hydroxyl, -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -OCH 2 OH, -OCH 2 CH 2 OH, -OCH 2 CH 2 CH 2 OH, -NHCH 2 OH, -NHCH 2 CH 2 OH, -NHCH 2 CH 2 CH 2 OH, -N(CH 3 )CH 2 OH, -N(CH 3 )CH 2 CH 2 OH, -N(CH 3 )CH 2 CH 2 CH 2 OH, -N(COCH 3 )CH 2 OH, - N(COCH 3 )CH 2 CH 2 OH, -N(COCH 3 )CH 2 CH 2 CH 2 OH, -N(SO 2 CH 3 )CH 2 OH, -N(SO 2 CH 3 )CH 2 OH
  • said R a is selected from the following groups:
  • the Ra is selected from the following groups:
  • the Q is selected from: -NR2- ,
  • the alkyl moiety is optionally substituted with one or more R 9 selected from: hydrogen, halogen, amino, cyano, carboxyl , C 1 -C 6 alkyl, C 1 -C 6 alkoxy or aromatic group; more preferably, the R 2 is selected from: R 9
  • the R n is selected from: hydroxyl-substituted C 1 -C 8 alkyl; more preferably, the R n is selected from: hydroxyl-substituted C 1 -C 4 alkyl; most preferably, the R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH or -CH 2 CH 2 CH 2 CH 2 OH.
  • the R n is selected from: hydroxyl, mercapto or amino-substituted C 1 -C 8 alkyl; more preferably, the R n is selected from: hydroxyl, mercapto or amino substituted C 1 -C 4 alkyl; most preferably, R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH.
  • the R m is selected from: hydrogen, -C 1 -C 4 alkyl.
  • the R 2 is further selected from: hydrogen, methyl, ethyl, -SO 2 CH 3 , -COCH 3 , -CO-isopropyl, -CO-cyclopropyl, isopropyl, Cyclopropyl, 2-methoxyethyl, 2-cyanoethyl, phenyl, naphthyl, benzyl, 2-phenylethyl, 1-naphthylmethyl, 2-naphthylethyl, -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH
  • each R 9 is independently selected from: hydrogen, R 13 , halogen, cyano, nitro, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, C 1 -C 6 alkoxy;
  • Each a1 is independently selected from: 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, each a2 is independently selected from: 0, 1, 2, 3, 4, 5, 6, 7, 8.
  • Each a3 is independently selected from: 0, 1, 2, 3, 4, 5, 6, 7.
  • Each a4 is independently selected from: 0, 1, 2, 3, 4, 5, 6.
  • Each a5 is independently selected. Selected from: 0, 1, 2, 3, 4, 5.
  • the R 13 is selected from: hydroxyl, R n , - OR n .
  • the R n is selected from: hydroxyl-substituted C 1 -C 8 alkyl; more preferably, R n is selected from: hydroxyl-substituted C 1 -C 4 alkyl; most preferably, R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH or -CH 2 CH 2 CH 2 CH 2 OH.
  • the R n is selected from: hydroxyl, mercapto or amino-substituted C 1 -C 8 alkyl; more preferably, the R n is selected from: hydroxyl, mercapto or amino substituted C 1 -C 4 alkyl; Most preferably, R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH.
  • the R m is selected from: hydrogen, -C 1 -C 4 alkyl.
  • the T and Z are independently selected from: chemical bond, carbonyl group, C 1 -C 6 alkylene group or C 3 -C 10 cycloalkylene group, the C 1 -C 6 alkylene group , C 3 -C 10 cycloalkylene is optionally substituted by one or more R 9 .
  • the T and Z are independently selected from: chemical bonds, carbonyl groups, methylene groups, 1,2-ethylene groups, 1,1-cyclopropylene groups or 2,2-propylene groups.
  • Methyl, 1,2-ethylene, and 1,1-cyclopropylene are optionally substituted by one or more R 9 .
  • the R 9 is selected from: hydrogen, R 13 , C 1 -C 6 alkyl or C 3 -C 8 cycloalkyl.
  • the R 13 is selected from: hydroxyl, R n , -OR n .
  • the R n is selected from: hydroxyl-substituted C 1 -C 8 alkyl; more preferably, R n is selected from: hydroxyl-substituted C 1 -C 4 alkyl; most preferably, the R n is selected from From: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, or -CH 2 CH 2 CH 2 CH 2 OH.
  • the R n is selected from: hydroxyl, mercapto or amino-substituted C 1 -C 8 alkyl; preferably, the R n is selected from: hydroxyl, mercapto or amino substituted C 1 -C 4 Alkyl group; more preferably, the R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH.
  • the U is selected from: C 1 -C 6 alkylene, C 3 -C 10 cycloalkylene, arylene or heteroarylene, the alkylene, cycloalkylene , arylene group and heteroarylene group are optionally substituted by one or more R 9 .
  • the U is selected from: C 2 -C 6 alkylene, C 5 -C 6 cycloalkylene, C 6 -C 10 arylene, 5-6 membered monocyclic heteroarylene, the Alkylene, cycloalkylene, arylene, heteroarylene are optionally substituted by one or more R 9 .
  • the U is selected from: 1,2-ethylene, 1,3-propylene, 1,4-butylene, 1,5-pentylene, 1,6-hexylene, 1,3 -Cyclopentylene, 1,3-cyclohexylene, 1,4-cyclohexylene, 1,2-phenylene, 1,3-phenylene, 1,4-phenylene, 2,5- Pyridinyl, 2,5-pyrimidinyl, 2,5-thiazolyl or 2,4-oxazolyl, the 1,2-ethylene, 1,3-propylene, 1,4 -Butylene, 1,3-cyclopentylene, 1,3-cyclohexylene, 1,4-cyclohexylene, 1,2-phenylene, 1,3-phenylene, 1,4-phenylene Phenyl, 2,5-pyridinylene, 2,5-pyrimidinyl, 2,5-thiazolylene, and 2,4-oxazolylene are optionally substituted by one or more R 9 .
  • the R 9 is selected from: hydrogen, R 13 , halogen, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, C 1 -C 6 alkoxy.
  • the R n is selected from: hydroxyl-substituted C 1 -C 8 alkyl; more preferably, R n is selected from: hydroxyl-substituted C 1 -C 4 alkyl; most preferably, R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH or -CH 2 CH 2 CH 2 CH 2 OH.
  • the R n is selected from: hydroxyl, mercapto or amino-substituted C 1 -C 8 alkyl; preferably, the R n is selected from: hydroxyl, mercapto or amino substituted C 1 -C 4 Alkyl group; more preferably, the R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH.
  • the R m is selected from: hydrogen, -C 1 -C 4 alkyl.
  • the R 9 is selected from: F, Cl, Br, cyano, methyl, ethyl, trifluoromethyl, methoxy, ethoxy, hydroxyl, -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -OCH 2 OH, -OCH 2 CH 2 OH, -OCH 2 CH 2 CH 2 OH, -NHCH 2 OH, -NHCH 2 CH 2 OH, -NHCH 2 CH 2 CH 2 OH, -N(CH 3 )CH 2 OH, -N(CH 3 )CH 2 CH 2 OH, -N(CH 3 )CH 2 CH 2 CH 2 OH, -N(COCH 3 )CH 2 OH, - N(COCH 3 )CH 2 CH 2 OH, -N(COCH 3 )CH 2 CH 2 CH 2 OH, -N(SO 2 CH 3 )CH 2 OH, -N(SO 2 CH 3 )CH 2 OH
  • said U is selected from the following groups:
  • the -TUZ- taken together form a group selected from:
  • the R 9 is selected from: hydrogen, C 1 -C 4 alkyl, R 13 ; more preferably, the R 9 is selected from: R 13 .
  • the R 13 is selected from: hydroxyl, R n , -OR n .
  • the base part is optionally substituted by one or more R 9 selected from: hydrogen, halogen, amino, cyano, carboxyl, C 1 -C 6 alkyl, C 1 -C 6 alkoxy or aryl ; More preferably, the
  • the R n is selected from: hydroxyl-substituted C 1 -C 8 alkyl; more preferably, the R n is selected from: hydroxyl-substituted C 1 -C 4 alkyl; most preferably, the R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH or -CH 2 CH 2 CH 2 CH 2 OH.
  • the R n is selected from: hydroxyl, mercapto or amino-substituted C 1 -C 8 alkyl; more preferably, the R n is selected from: hydroxyl, mercapto or amino substituted C 1 -C 4 alkyl; most preferably, R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH.
  • the R m is selected from: hydrogen, -C 1 -C 4 alkyl.
  • the R 2 is further selected from: hydrogen, methyl, ethyl, -SO 2 CH 3 , -COCH 3 , -CO-isopropyl, -CO-cyclopropyl, isopropyl, Cyclopropyl, 2-methoxyethyl, 2-cyanoethyl, phenyl, naphthyl, benzyl, 2-phenylethyl, 1-naphthylmethyl, 2-naphthylethyl, -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH
  • the R n is selected from: hydroxyl-substituted C 1 -C 8 alkyl; more preferably, R n is selected from: hydroxyl-substituted C 1 -C 4 alkyl; most preferably, R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH or -CH 2 CH 2 CH 2 CH 2 OH.
  • the R n is selected from: hydroxyl, mercapto or amino-substituted C 1 -C 8 alkyl; more preferably, the R n is selected from: hydroxyl, mercapto or amino substituted C 1 -C 4 alkyl; most preferably, R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH.
  • each of R 3 to R 5 is independently selected from: hydrogen, R 13 , halogen, cyano, amino, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkylthio group, C 1 -C 6 alkylamino group, di(C 1 -C 6 alkyl)amine group, C 3 -C 8 cycloalkyl group, the alkyl group, alkoxy group, alkylamino group , alkylthio and cycloalkyl are optionally substituted by 1-3 groups respectively selected from halogen, C 1 -C 3 alkyl or C 1 -C 3 alkoxy; when R 3 -R 5 is multiple In this case, any two adjacent ones can be combined to form a ring.
  • said R 3 is selected from: R 13 .
  • the R n is selected from: hydroxyl-substituted C 1 -C 8 alkyl; more preferably, R n is selected from: hydroxyl-substituted C 1 -C 4 alkyl; most preferably, R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH or -CH 2 CH 2 CH 2 CH 2 OH.
  • the R n is selected from: hydroxyl, mercapto or amino-substituted C 1 -C 8 alkyl; more preferably, the R n is selected from: hydroxyl, mercapto or amino substituted C 1 -C 4 alkyl; most preferably, R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH.
  • R m is selected from: hydrogen, -C 1 -C 4 alkyl.
  • the R 4 to R 5 are each independently selected from: hydrogen, halogen, cyano, C 1 -C 4 alkyl, C 1 -C 4 haloalkyl.
  • the R 6 is selected from: hydrogen, C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl, and the alkyl and cycloalkyl are optionally selected from 1 to 3 respectively.
  • R 23 is selected from: hydrogen, C 1 -C 4 alkyl, so The C 1 -C 4 alkyl group is optionally substituted by 1 to 3 groups respectively selected from hydroxyl and amino groups.
  • R 23 is selected from: hydrogen, C 1 -C 4 alkyl, and the C 1 -C 4 alkyl is optionally substituted by 1-3 groups respectively selected from hydroxyl and amino.
  • the R 6 is selected from: hydrogen, C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl, and the alkyl and cycloalkyl are optionally selected from 1 to 3 respectively.
  • the R 6 is selected from: hydrogen, C 1 -C 4 alkyl, C 1 -C 4 haloalkyl.
  • the R a is selected from the following groups:
  • the Ra is further preferably the following group:
  • Q is selected from: -NR 2 -and
  • R n is selected from: hydroxyl-substituted C 1 -C 8 alkyl; preferably, R n is selected from: hydroxyl, mercapto or amino-substituted C 1 -C 8 alkyl;
  • R m is selected from: hydrogen and -C 1 -C 4 alkyl
  • T and Z are independently selected from: chemical bond, carbonyl group, methylene group, 1,2-ethylene group, 1,1-cyclopropylene group or 2,2-propylene group;
  • R 3 to R 6 are each independently selected from: hydrogen and C 1 -C 6 alkyl
  • X is CH 2 ;
  • a is selected from: 0, 1, 2, 3, 4 or 5;
  • b is selected from: 0, 1, 2, 3, 4 or 5;
  • n is selected from: 0, 1 or 2;
  • n is selected from: 0, 1 or 2.
  • the R a is selected from the following groups:
  • the Ra is further preferably the following group:
  • Q is selected from: -NR 2 -and
  • R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH and -CH 2 CH 2 CH 2 CH 2 OH; preferably, the R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH;
  • R m is hydrogen
  • -TUZ- is not C 1-4 alkylene
  • R 3 to R 6 are each independently hydrogen
  • X is CH 2 ;
  • the R a is selected from the following groups:
  • the Ra is further preferably the following group:
  • Q is selected from: -NR 2 -and
  • R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 CH 2 SH;
  • the R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH and -CH 2 CH 2 CH 2 CH 2 OH;
  • R m is hydrogen
  • R 3 to R 6 are each independently hydrogen
  • X is CH 2 ;
  • the Ra is selected from the following groups:
  • the Ra is further preferably the following group:
  • the structural unit represented by formula III is a structural unit represented by the following formula III-A
  • R 1 , R 3 , R 4 , R 5 , R 6 , Q, T, U, Z, X, Y, m and n are as defined above.
  • D is selected from
  • the antibody drug conjugate structure is shown in Formula IV,
  • Tb, S, L 1 , L 2 , R x1 , R y1 , D and q are defined as described in any scheme of this disclosure.
  • antibodies or antigen-binding fragments thereof suitable for use in the ADCs of the invention are anti-PSMA antibodies or antigen-binding fragments thereof.
  • an antibody or antigen-binding fragment thereof suitable for use in the ADC of the invention is an antibody portion that specifically binds PSMA, such as the PSMA antibodies disclosed by Progenics, or the antibodies in its PSMA ADC, such as those in US9242012B2 or WO2021/190583A1 PSMA antibodies or antigen-binding fragments thereof in ADCs or conjugates disclosed in.
  • PSMA antibodies or antigen-binding fragments thereof suitable for use in the ADC of the invention comprise known antibodies that specifically bind PSMA, such as the PSMA antibodies disclosed by Progenics, or antibody portions thereof in PSMA ADCs, such as those in US9242012B2 Or 1, 2, 3, 4, 5, or 6 CDRs of the antibody in the ADC or conjugate disclosed in WO2021/190583A1.
  • the antigen-binding region comprises an antibody known to specifically bind PSMA, such as the PSMA antibodies disclosed by Progenics, or an antibody in a PSMA ADC thereof, such as the ADC or conjugate disclosed in US9242012B2 or WO2021/190583A1
  • the 1, 2 and 3 heavy chain variable region CDRs of the antibody namely HCDR1, HCDR2 and HCDR3.
  • the antigen-binding region comprises an antibody known to specifically bind PSMA, such as the PSMA antibodies disclosed by Progenics, or the antibodies in its PSMA ADC, such as those disclosed in US9242012B2 Or 1, 2 and 3 light chain variable region CDRs of the antibody in the ADC or conjugate disclosed in WO2021/190583A1, namely LCDR1, LCDR2 and LCDR3.
  • an antibody to PSMA such as a PSMA antibody disclosed by Progenics, or an antibody in its PSMA ADC, such as an ADC disclosed in US9242012B2 or WO2021/190583A1, or the three heavy chain variable regions of an antibody in a conjugate CDRs and 3 light chain variable region CDRs.
  • the antigen-binding region comprises an antibody known to specifically bind PSMA, such as the PSMA antibodies disclosed by Progenics, or an antibody in a PSMA ADC thereof, such as the ADC or conjugate disclosed in US9242012B2 or WO2021/190583A1
  • PSMA antibodies disclosed by Progenics
  • ADC an antibody in a PSMA ADC thereof, such as the ADC or conjugate disclosed in US9242012B2 or WO2021/190583A1
  • the heavy chain variable region of the antibody, and/or the light chain variable region is provided.
  • the antigen-binding region comprises an antibody known to specifically bind PSMA, such as the PSMA antibodies disclosed by Progenics, or an antibody in a PSMA ADC thereof, such as the ADC or conjugate disclosed in US9242012B2 or WO2021/190583A1
  • PSMA antibodies disclosed by Progenics
  • ADC an antibody in a PSMA ADC thereof, such as the ADC or conjugate disclosed in US9242012B2 or WO2021/190583A1
  • the PSMA antibody or antigen-binding fragment thereof is an antibody or antigen-binding fragment thereof containing the following CDRs: HCDR1 shown in SEQ ID No. 7, HCDR2 shown in SEQ ID No. 8, SEQ ID No. HCDR3 shown in .9, LCDR1 shown in SEQ ID No.10, LCDR2 shown in SEQ ID No.11, LCDR3 shown in SEQ ID No.12.
  • an antibody or antigen-binding fragment thereof that specifically binds PSMA comprises a heavy chain variable region (VH), wherein the VH
  • (iii) Comprises one or more (preferably no more than 10, more preferably no more than 5, 4, 3, 2, 1) amino acid changes (preferably amino acid substitutions) compared to the amino acid sequence of SEQ ID NO: 1 , more preferably amino acid conservative substitution) amino acid sequence consists of the amino acid sequence, preferably, the amino acid changes do not occur in the CDR region.
  • an antibody or antigen-binding fragment thereof that specifically binds PSMA comprises a light chain variable region (VL), wherein the VL
  • (iii) Comprises one or more (preferably no more than 10, more preferably no more than 5, 4, 3, 2, 1) amino acid changes (preferably amino acid substitutions) compared to the amino acid sequence of SEQ ID NO:3 , more preferably amino acid conservative substitution) amino acid sequence consists of the amino acid sequence, preferably, the amino acid changes do not occur in the CDR region.
  • the PSMA antibody or antigen-binding fragment thereof contains the VH shown in SEQ ID No. 1 and the VL shown in SEQ ID No. 3.
  • the PSMA antibody or antigen-binding fragment thereof further comprises a heavy chain constant region and/or a light chain constant region, wherein
  • (iii) Comprises one or more (preferably no more than 10, more preferably no more than 5, 4, 3, 2, 1) amino acid changes (preferably amino acid substitutions) compared to the amino acid sequence of SEQ ID NO:2 , more preferably the amino acid sequence consisting of the amino acid sequence consisting of the amino acid conservative substitution);
  • (iii) Comprises one or more (preferably no more than 10, more preferably no more than 5, 4, 3, 2, 1) amino acid changes (preferably amino acid substitutions) compared to the amino acid sequence of SEQ ID NO:4 , more preferably the amino acid sequence consists of the amino acid sequence consisting of the amino acid conservative substitution).
  • the PSMA antibody or antigen-binding fragment thereof contains: VH shown in SEQ ID No. 1 and CH shown in SEQ ID No. 2, and VL shown in SEQ ID No. 3 and CL shown in SEQ ID No.4.
  • the PSMA antibody or antigen-binding fragment thereof contains: the heavy chain shown in SEQ ID No. 5 and the light chain shown in SEQ ID No. 6.
  • the antibody drug conjugate (ADC) is selected from the following:
  • q is selected from 1-10, such as 2-8.
  • the antibody drug conjugate has a drug to antibody ratio (average DAR) of any value selected from the following range: 1.0-20.0, 1.0-18.0, 1.0-16.0, 1.0-10.0, 2.0 -14.0, 3.0-12.0, 4.0-10.0, 5.0-9.0, 6.0-8.0 or 2.0-8.0.
  • a drug to antibody ratio average DAR
  • the average DAR is any value within 2+/-0.4, 4+/-0.4, 6+/-0.4, or 8+/-0.4.
  • the average DAR is about 2.0, 4.0, 6.0, 8.0, 10.0, or 12.0.
  • the present disclosure provides a drug linker conjugate represented by Formula V,
  • L' is a linker precursor
  • D is defined as described in any scheme of this disclosure.
  • a drug linker conjugate represented by Formula VI, its stereoisomer, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate is provided,
  • L 1 is defined as described in any aspect of this disclosure, and L 1 is not When , Lg is the leaving group when reacting with the antibody; L 2 , L 3 , L 4 and D are defined as described in any scheme of this disclosure;
  • Lg-L 1 is L 2 , L 3 , L 4 and D are as defined in any aspect of this disclosure.
  • Lg is selected from halogen, sulfone, tertiary amine (Me 3 N + , Et 3 N + ), diazonium, -OMs, MeSO 2 -, CF 3 SO 3 -, p Tosyl.
  • Lg is selected from F, Cl, Br, MeSO 2 -; more preferably; Lg is MeSO 2 -.
  • a drug linker conjugate represented by Formula VII, its stereoisomer, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate is provided,
  • Lg, L 1 , L 2 , R x1 , R y1 and D are as described in any aspect of this disclosure.
  • the drug linker conjugate is selected from the following:
  • R 1 , Q, T, U, Z, Y, X, R 3 , R 4 , R 5 , R 6 , m, and n are as described in any aspect of this disclosure.
  • the compound of Formula VIII has the structure shown in Formula VIII-A below
  • R 1 , R 3 , R 4 , R 5 , R 6 , Q, T, U, Z, X, Y, m and n are as defined above.
  • the structure of the protein-degrading bioactive compound is as follows:
  • a method for preparing an antibody-drug conjugate represented by Formula I includes: performing a coupling reaction between Tb and a drug linker conjugate represented by Formula V in a solvent to form C-S key steps.
  • the molar ratio of the Tb to the drug linker conjugate represented by Formula V is 1:(1-20), such as 1:2, 1:4, 1:6, 1:8, 1:10, 1:12, 1:14, 1:16, 1:18, 1:(10-20), 1:(12-20), 1:(14-20), 1:(16-20 ) or 1:(18-20).
  • the coupling reaction is performed in water and/or organic solvents.
  • the organic solvent is selected from one of N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, N-methylpyrrolidone or any of them. combination.
  • the method further includes the step of purifying the coupling product.
  • the coupling product is purified by chromatography.
  • the chromatography method includes one or more of ion exchange chromatography, hydrophobic chromatography, reversed phase chromatography, or affinity chromatography.
  • the method is carried out at -20-100°C, such as 0-50°C, preferably room temperature.
  • the disclosure provides a method for preparing a drug linker conjugate
  • the present disclosure provides a pharmaceutical composition
  • a pharmaceutical composition comprising the aforementioned antibody drug conjugate, or a stereoisomer of the antibody drug conjugate, a prodrug thereof, a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable salt thereof.
  • the present disclosure provides the aforementioned antibody drug conjugate, or the stereoisomer of the antibody drug conjugate, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate. or the aforementioned drug linker conjugate, its stereoisomer, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate; or the aforementioned pharmaceutical composition is used in the preparation of Use in medicines for the treatment and/or prevention of diseases associated with abnormal cellular activity, such as cancer diseases.
  • the present disclosure also provides methods for preventing and/or treating diseases related to abnormal cell activity (such as cancer diseases), which include administering to an individual the aforementioned antibody drug conjugate, or a stereoisomer of the antibody drug conjugate. body, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate; or the aforementioned drug linker conjugate, its stereoisomer, its prodrug, its pharmaceutically acceptable salt or pharmaceutically acceptable solvate thereof; or the aforementioned pharmaceutical composition.
  • diseases related to abnormal cell activity such as cancer diseases
  • the present disclosure also provides the aforementioned antibody drug conjugate, or the stereoisomer of the antibody drug conjugate, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate; or Such as the aforementioned drug linker conjugates, their stereoisomers, their prodrugs, their pharmaceutically acceptable salts or their pharmaceutically acceptable solvates; or the aforementioned pharmaceutical compositions, which are used to treat and /or prevent diseases, such as those associated with abnormal cellular activity (e.g., cancer diseases).
  • diseases such as those associated with abnormal cellular activity (e.g., cancer diseases).
  • the cancer disease is a solid tumor or a hematological tumor.
  • the cancer disease is selected from the group consisting of esophageal cancer (eg, esophageal adenocarcinoma and esophageal squamous cell carcinoma), brain tumors, lung cancer (eg, small cell lung cancer and non-small cell lung cancer), squamous cell carcinoma , bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, colorectal cancer, liver cancer, renal cancer, urothelial cancer, solid tumors, non-Hodge cancer Gold lymphoma, central nervous system tumors (such as glioma, glioblastoma multiforme, glioma, or sarcoma), prostate cancer, or thyroid cancer.
  • esophageal cancer eg, esophageal adenocarcinoma and esophageal squamous cell carcinoma
  • brain tumors eg, e
  • protein degrader refers to a class of compounds, preferably small molecule compounds, that can induce and ubiquitylate a protein of interest through E3 ligases and then degrade it through the proteasome.
  • Myc protein degrading agent refers to a protein degrading agent that has the ability to degrade Myc family proteins, such as a protein degrading agent that has the ability to degrade c-Myc protein and a protein degrading agent that has the ability to degrade n-Myc protein.
  • c-Myc protein degrading agent refers to a protein degrading agent that has the ability to degrade c-Myc protein.
  • protein degradation targeting chimera generally includes three parts: an E3 ubiquitin ligase ligand and a target protein ligand, two active ligands through a specially designed Linker structure Linked together, by recruiting E3 ubiquitin ligase to the target protein, it ubiquitinates the target protein and then degrades the target protein.
  • MLD molecular glue degrader
  • the term "pharmaceutically acceptable salt” means a salt that retains the biological effects and properties of the antibody-drug conjugate or drug-linker conjugate of the invention, and the salt is biologically or Other aspects are not undesirable.
  • the conjugates of the present invention may exist in the form of their pharmaceutically acceptable salts, including acid addition salts and base addition salts.
  • a pharmaceutically acceptable acid addition salt means that the conjugate in the present invention is formed with an organic or inorganic acid.
  • Salts, organic or inorganic acids include but are not limited to hydrochloric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, nitric acid, perchloric acid, acetic acid, oxalic acid, maleic acid, fumaric acid, tartaric acid, benzenesulfonic acid, formazan Sulfonic acid, salicylic acid, succinic acid, citric acid, lactic acid, propionic acid, benzoic acid, p-toluenesulfonic acid, malic acid, etc.
  • Pharmaceutically acceptable base addition salts refer to salts formed by the conjugate in the present invention and organic or inorganic bases, including but not limited to alkali metal salts, such as lithium, sodium or potassium salts; alkaline earth metal salts, such as calcium or Magnesium salts; organic base salts, such as ammonium salts formed with organic bases containing N groups.
  • alkali metal salts such as lithium, sodium or potassium salts
  • alkaline earth metal salts such as calcium or Magnesium salts
  • organic base salts such as ammonium salts formed with organic bases containing N groups.
  • compositions can be obtained using standard procedures well known in the art, for example, by reacting a sufficient amount of a basic compound with a suitable acid that provides a pharmaceutically acceptable anion.
  • stereoisomer means an isomer formed due to at least one asymmetric center.
  • compounds with one or more (e.g., one, two, three or four) asymmetric centers they can give rise to racemic mixtures, single enantiomers, diastereomeric mixtures and individual of diastereomers.
  • Certain individual molecules may also exist as geometric isomers (cis/trans).
  • compounds of the present invention may exist as mixtures of two or more structurally distinct forms in rapid equilibrium (often referred to as tautomers).
  • tautomers include keto-enol tautomers, phenol-ketone tautomers, nitroso-oxime tautomers, and imine-enamine tautomers. wait. It is to be understood that the scope of the present disclosure encompasses all such products in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99 %) isomers or mixtures thereof.
  • the carbon-carbon bonds of the compounds of the invention may be depicted in this disclosure using solid lines (—), solid wedges, or dashed wedges.
  • the use of a solid line to depict a bond to an asymmetric carbon atom is intended to indicate that all possible stereoisomers at that carbon atom are included (eg, a specific enantiomer, a racemic mixture, etc.).
  • the use of solid or imaginary wedges to depict bonds to asymmetric carbon atoms is intended to demonstrate that the stereoisomers shown exist. When present in a racemic mixture, solid and imaginary wedges are used to define relative stereochemistry rather than absolute stereochemistry.
  • the compounds of the present invention are intended to exist as stereoisomers (which includes cis and trans isomers, optical isomers (e.g., R and S enantiomers), diastereomers, They exist in the form of geometric isomers, rotamers, conformational isomers, atropisomers and mixtures thereof).
  • the compounds of the present invention may exhibit more than one type of isomerism and consist of mixtures thereof (eg, racemic mixtures and pairs of diastereoisomers).
  • the present disclosure also includes all pharmaceutically acceptable isotopic compounds that are identical to the compounds of the invention except that one or more atoms are assigned the same atomic number but an atomic mass or mass number that is different from the atomic mass or mass that predominates in nature Atomic substitution of numbers.
  • isotopes suitable for inclusion in the compounds of the present disclosure include, but are not limited to, isotopes of hydrogen (e.g. 2 H, 3 H); isotopes of carbon (e.g. 11 C, 13 C and 14 C); isotopes of chlorine (e.g. 36 Cl); isotopes of fluorine (e.g.
  • Iodine isotopes such as 123 I and 125 I
  • Nitrogen isotopes such as 13 N and 15 N
  • Oxygen isotopes such as 15 O, 17 O and 18 O
  • Phosphorus isotopes such as 32 P
  • isotopes of sulfur e.g. 35 S.
  • the compounds of the present disclosure may exist in the form of solvates, preferably hydrates, wherein the compounds of the present disclosure comprise a polar solvent as a structural element of the crystal lattice of the compound, in particular such as water, methanol or ethanol.
  • a polar solvent as a structural element of the crystal lattice of the compound, in particular such as water, methanol or ethanol.
  • the amount of polar solvent, especially water, may be present in stoichiometric or non-stoichiometric ratios.
  • metabolites of the compounds of the invention ie substances formed in the body upon administration of the compounds of the invention. Such products may result, for example, from oxidation, reduction, hydrolysis, amidation, deamidation, esterification, delipidation, enzymatic hydrolysis, etc. of the administered compound.
  • the invention includes metabolites of the compounds of the invention, including compounds prepared by contacting a compound of the invention with a mammal for a time sufficient to produce metabolites thereof.
  • the present disclosure further includes within its scope prodrugs of the compounds of the invention.
  • prodrugs will be functional group derivatives of the compound that are readily converted in vivo to the desired therapeutically active compound. Therefore, in these instances, the term “administering" as used in the treatment methods of the present disclosure shall include the treatment of various diseases or conditions with prodrug forms of one or more of the claimed compounds, but in The prodrug form is converted in vivo to the compound described above upon administration to an individual.
  • “Design of Prodrug” ed. H. Bundgaard, Elsevier, 1985, general methods for selecting and preparing suitable prodrug derivatives are described.
  • the pharmaceutical excipients refer to the excipients and additives used in the production of drugs and preparation of prescriptions. They refer to excipients and additives that have been reasonably evaluated in terms of safety in addition to active ingredients and are included in pharmaceutical preparations. substance in.
  • pharmaceutical excipients also have important functions such as solubilization, dissolution, and sustained and controlled release. They are important ingredients that may affect the quality, safety, and effectiveness of drugs. According to their origin, they can be divided into natural products, semi-synthetic products and fully synthetic products.
  • solvents can be divided into: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, Glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesive agents, antioxidants, chelating agents, penetration enhancers, pH regulators, buffers, plasticizers, surface active agents Agents, foaming agents, defoaming agents, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants and deflocculants, filter aids, release retardants, etc.; can be divided according to their route of administration for mouth Oral administration, injection, mucous membrane, transdermal or topical administration, nasal or oral inhalation administration and ocular administration, etc.
  • the same pharmaceutical excipients can be used in pharmaceutical preparations with different
  • the pharmaceutical composition can be prepared into various suitable dosage forms according to the route of administration.
  • suitable dosage forms for example, tablets, capsules, granules, oral solutions, oral suspensions, oral emulsions, powders, tinctures, syrups, injections, suppositories, ointments, creams, pastes, ophthalmic preparations, pills, implants agents, aerosols, powder mist, sprays, etc.
  • the pharmaceutical composition or suitable dosage form may contain 0.01 mg to 1000 mg of the compound of the present disclosure (including conjugates) or a pharmaceutically acceptable salt thereof, suitably 0.1 mg to 800 mg, preferably 0.5-500 mg. , preferably contains 0.5 to 350 mg, particularly preferably 1 to 250 mg.
  • the pharmaceutical composition can be administered in the form of injection, including injection liquid, sterile powder for injection and concentrated solution for injection.
  • carriers and solvents that can be used include water, Ringer's solution and isotonic sodium chloride solution.
  • sterile fixed oils may be used as solvents or suspending media, such as mono- or diglycerides.
  • the pharmaceutical composition may be administered in the form of an infusion.
  • treatment generally refers to obtaining a desired pharmacological and/or physiological effect.
  • the effect may be prophylactic in terms of completely or partially preventing the disease or its symptoms; and/or may be therapeutic in terms of partially or completely stabilizing or curing the disease and/or side effects due to the disease.
  • treatment encompasses any treatment of a disease in a patient, including: (a) preventing the disease or symptoms in a patient who is susceptible to the disease or condition but has not yet been diagnosed with the disease; (b) suppressing the symptoms of the disease , i.e., arresting its progression; or (c) alleviating the symptoms of a disease, i.e., causing regression of the disease or symptoms.
  • the term "individual” includes humans or non-human animals.
  • Exemplary human subjects include human subjects (referred to as patients) suffering from a disease, such as those described in this disclosure, or normal subjects.
  • non-human animals in this disclosure includes all vertebrate animals, such as non-mammals (e.g., birds, amphibians, reptiles) and mammals, such as non-human primates, domestic animals, and/or domesticated animals (e.g., sheep, dogs, cats, cows, pigs, etc.).
  • the term "effective dose” refers to an amount of an antibody-drug conjugate, drug-linker conjugate, compound or composition that when administered will alleviate to a certain extent one or more symptoms of the condition being treated. .
  • the terms "antibody drug conjugate” and "ADC” refer to a substance obtained by connecting a biologically active compound fragment (drug molecule) to an antibody or its antigen-binding fragment portion.
  • the bioactive compound fragment is linked to the targeting moiety via a linker.
  • the linker can be cleaved in a specific environment (such as an intracellular low pH environment) or under a specific action (such as the action of a lysosomal protease), thereby allowing the bioactive compound (such as a c-Myc protein degrader) fragment to interact with the target. Separation into parts or antibodies or antigen-binding fragments thereof.
  • the linker comprises a cleavable or non-cleavable units such as peptides or disulfide bonds.
  • the bioactive compound fragment is directly connected to the targeting moiety or antibody or antigen-binding fragment thereof by a covalent bond, which can be broken under specific circumstances or effects, thereby rendering the bioactive compound The fragment is partially separated from the antibody or antigen-binding fragment thereof.
  • the 1-position of L is connected to Tb through an S atom
  • the 1-position of L is responsible for opening the disulfide bond (for example, the disulfide bond can be opened by reducing the disulfide bond through the reducing agent TCEP).
  • the disulfide bond which generates the sulfhydryl group -SH), is connected to the sulfhydryl group contained in Tb (such as an antibody) itself.
  • the -S- between L and Tb is not an additional external sulfur atom.
  • -S- is not another external sulfur atom, but the thiol group and L contained in Tb itself after opening the disulfide bond, for example 1 bits are connected to form -S-.
  • unsubstituted means that the referenced group is not substituted with a substituent, it being understood that the referenced group will contain an appropriate number of hydrogen atoms to comply with valence rules.
  • the term "substituted by” refers to the independent presence of one or more substituents at any carbon (or nitrogen) position of the molecule, preferably 1-5 (such as 1, 2, 3, 4 or 5 ) substituent, most preferably 1-3 substituents, the substituent may be: hydroxyl, mercapto, carboxyl, cyano, nitro, halogen (preferably, 1, 2 or 3 halogen, especially on the alkyl group, especially on the alkyl group Methyl, for example trifluoromethyl), alkyl (preferably C 1 -C 10 , more preferably C 1 -C 6 ), haloalkyl, alkenyl, alkynyl, cycloalkyl, aryl (especially phenyl ), heteroaryl, heterocyclyl, alkoxy (preferably C 1 -C 6 alkoxy), aryloxy (preferably phenoxy), thioether (C 1 -C 6 alkylthio or arylthio) Such as
  • alkyl refers to a straight-chain, branched-chain fully saturated hydrocarbon group that may be optionally substituted, preferably C 1 -C 10 , more preferably C 1 -C 8 , C 1 -C 6 , or C 1 -C 4 alkyl.
  • alkyl groups are methyl, ethyl, Propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl or n-decyl, etc.
  • alkylene refers to a divalent group formed by further removing one hydrogen atom from an alkyl group.
  • cycloalkyl refers to a cyclic alkyl group that may be optionally substituted, preferably C 3 -C 20 , more preferably C 3 -C 15 , or C 3 -C 8 cycloalkyl.
  • Examples of cycloalkyl groups are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexyl or cycloheptyl, among others.
  • cycloalkylene refers to a divalent group formed by further removing one hydrogen atom from a cycloalkyl group.
  • bridged cyclic group refers to a cyclic structure formed by two or more cyclic structures that may be optionally substituted and share two non-adjacent carbon atoms with each other, preferably C 5 -C 15 , more preferably C 6 -C 12 , or C 7 -C 10 bridged ring group.
  • Examples of bridged cyclic groups are bicyclo[1.1.1]pentyl, norbornyl, adamantyl and the like.
  • alkynyl refers to a linear, branched or cyclic C 2 -C 10 (preferably C 2 -C 8 ) hydrocarbon group containing at least one C ⁇ C bond.
  • aryl refers to C 6 -C 16 having a single ring (such as phenyl) or a condensed ring (such as naphthyl, anthracenyl, phenanthrenyl, fluorenyl, etc.) that may be optionally substituted.
  • Aromatic hydrocarbon groups preferably C 6 -C 10 aromatic hydrocarbon groups.
  • arylene refers to a divalent group formed by further removing one hydrogen atom from an aryl group.
  • heteroaryl refers to an optionally substituted 5- containing one or more (eg, 1, 2, 3, or 4) heteroatoms selected from N, O, S, or P.
  • a 16-membered aromatic group is preferably a 5- to 10-membered aromatic group, and more preferably a 5- to 6-membered aromatic group.
  • heteroaryl groups include imidazolyl, pyrazolyl, triazolyl, tetrazolyl, pyrrolyl, furyl, thienyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyridyl Azinyl, pyrimidinyl, pyridazinyl, indolyl, azaindolyl (such as 7-azaindolyl), benzimidazolyl, benzopyrazolyl, benzofuranyl, benzothienyl , benzothiazolyl, dibenzofuranyl, dibenzothienyl, quinolyl, isoquinolinyl, naphthyridinyl, carbazolyl, azacarbazolyl (such as 1-azacarbazolyl, 2-azacarbazolyl, 1,8-diazacarbazolyl), indolaziny
  • heteroaryl refers to a bivalent group formed by further removing one hydrogen atom from a heteroaryl group.
  • heterocyclyl refers to an optionally substituted group containing one or more (eg, 1, 2, 3 or 4) selected from N, O, S, SO, SO 2 or P Partially unsaturated or completely unsaturated 3-20-membered cyclic group of heteroatoms, preferably 3-10-membered cyclic group, more preferably 3-6-membered cyclic group; more preferably 5-6-membered cyclic group group; the heterocyclic group contains 1-19 carbon atoms, preferably 2-10 carbon atoms, and more preferably 3-5 carbon atoms.
  • heterocyclyl examples include: aziridinyl, oxetanyl, azetidinyl, oxetanyl, 1,4-benzodioxanyl, 1,3-benzo Dioxolyl, dihydroimidazolyl, dihydropyranyl, dihydrofuryl, dioxanyl, ethyleneureido, 1,3-dioxolyl, 1, 3-dioxanyl, 1,4-dioxanyl, imidazolinyl, indolinyl, morpholinyl, pyridone, 2-pyrrolidone, piperazinyl, homopiperazinyl, piperidinyl, homo Piperidinyl, phthalimide, succinimide, pyrazinyl, pyrazolinyl, pyrrolinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydroquinolinyl, tetra
  • heterocyclylene refers to a bivalent group formed by further removing one hydrogen atom from the heterocyclyl group.
  • heterocycloalkyl refers to a fully saturated heterocyclyl group.
  • fused arylcycloalkyl refers to a group formed by the fusion of the above-mentioned aryl group and cycloalkyl group.
  • fused arylheterocyclyl refers to a group formed by the fusion of the above-mentioned aryl group and heterocyclyl.
  • fused heteroarylcycloalkyl refers to a group formed by the fusion of the above-mentioned heteroaryl and cycloalkyl.
  • fused heteroarylheterocyclyl refers to a group formed by the fusion of the above-mentioned heteroaryl and heterocyclyl.
  • aryl-alkyl means that the above-mentioned aryl group and alkyl group are connected through a single bond, and other parts are connected through the alkyl group.
  • heterocycle refers to an optionally substituted heterocyclic ring containing one or more (eg, 1, 2, 3, or 4) selected from N, O, S, SO, SO 2, or P Atomically partially unsaturated or fully unsaturated 3-20-membered cyclic groups, preferably 4-10-membered cyclic groups, 3-7-membered cyclic groups, more preferably 3-6-membered cyclic groups, including But it is not limited to pyrrolidine, dihydropyrrolidine, tetrahydrofuran, piperidine, piperazine, tetrahydropyran, pyrrolidone and other rings.
  • the term "carbocyclic ring” refers to a partially unsaturated or fully unsaturated 3-10-membered cyclic group in which all ring atoms are carbon atoms, preferably a 3-7-membered cyclic group, and more preferably a 3-6-membered cyclic group. cyclic group.
  • the carbon atoms in the cyclic structure can be oxo-substituted, including but not limited to cyclopropane, cyclobutane and other rings.
  • halogen refers to F, Cl, Br, I.
  • drug refers to a substance that inhibits or prevents the function of a cell and/or causes cell death or destruction.
  • linker refers to a fragment that connects a biologically active compound fragment (drug molecule) to an antibody moiety.
  • the linker prior to being attached to the antibody or antigen-binding fragment thereof (i.e., the linker precursor), has functional groups that can form bonds with functional groups of the antibody or antigen-binding fragment thereof.
  • the "... bioactive compound fragment” refers to the antibody-drug conjugate (or antibody-drug conjugate (ADC)) known in the art, which is used in tumors.
  • ADC antibody-drug conjugate
  • biologically active drugs such as small molecule cytotoxic drugs, which include groups after losing one atom or atomic group
  • derivatives thereof For example, its precursor part (fragment or group).
  • drug do not only refer to “drugs” that have been approved by the medical regulatory authorities, but also include any compounds with potential therapeutic biological activity in clinical practice, or in R&D and academic research.
  • the biologically active compound includes the Myc protein degrading agent of the present invention.
  • linker unit is a component of an antibody-drug conjugate or a drug-linker conjugate or a linker, and its function is to connect the antibody or antigen-binding fragment thereof that binds to the target with the antibody-drug conjugate. connected to the rest.
  • the linker unit can connect the Tb unit to L 2. Specific examples include but are not limited to (where 1 position is connected to the antibody or its antigen-binding fragment, and 2 position is connected to L 2 or L 3 ):
  • linking unit is a component of an antibody-drug conjugate or a drug-linker conjugate or a linker, which functions to combine the linker unit with an amino acid residue or consists of 2-10 amino acid residues. composed of short peptides.
  • the presence of the connecting unit can connect L 1 to L 3 . Specific examples include but are not limited to (where 1 is connected to the connector unit and 2 is connected to L 3 ):
  • spacer unit is a component of an antibody-drug conjugate or a drug-linker conjugate or a linker, which is used to bind the Myc protein degrader fragment to an amino acid residue or consists of 2-10 A short peptide consisting of amino acid residues, or which may provide additional structural components to further facilitate the release of the Myc protein-degrading agent from the remainder of the antibody drug conjugate.
  • the presence of a spacer unit can connect D to L 3 . Specific examples include but are not limited to (where 1 bit is connected to L 3 and 2 bits are connected to D):
  • antibody is taken in its broadest interpretation and includes intact monoclonal antibodies, polyclonal antibodies, and multispecific antibodies (e.g., bispecific antibodies) formed from at least two intact antibodies, as long as they have the required required biological activity.
  • antibody and “immunoglobulin” are used interchangeably.
  • PSMA antibody refers to an antibody that is capable of binding (human) PSMA with sufficient affinity such that the antibody can be used as a therapeutic targeting (human) PSMA.
  • the (human) PSMA antibody binds (human) PSMA with high affinity in vitro or in vivo.
  • the binding is measured, for example, by radioimmunoassay (RIA), biofilm thin layer interferometry (BLI), MSD assay, or surface plasmon resonance (SPR), or flow cytometry.
  • antibody fragment includes a portion of an intact antibody.
  • the antibody fragment is an antigen-binding fragment.
  • Antigen-binding fragment refers to a molecule distinct from an intact antibody that contains a portion of an intact antibody and binds the antigen to which the intact antibody binds.
  • antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2; dAb (domain antibody); linear antibodies; single chain antibodies (such as scFv); single domain antibodies such as VHH ; Diavalent antibodies or fragments thereof; or camelid antibodies.
  • the term "monoclonal antibody” refers to an antibody derived from a population of antibodies that is substantially homogeneous, that is, the individual antibodies that make up the population are identical except for the possible presence of a small number of natural mutations.
  • Monoclonal antibodies have high specificity for one determinant (epitope) of an antigen, whereas polyclonal antibodies contain different antibodies directed against different determinants (epitope).
  • epitopope epitope
  • the advantage of monoclonal antibodies is that they can be synthesized without contamination from other antibodies.
  • the modifier "monoclonal” here indicates that the antibody is characterized by being derived from a substantially homogeneous population of antibodies and should not be construed as requiring that it be prepared by a specific method.
  • monoclonal antibodies also specifically include chimeric antibodies, that is, a part of the heavy chain and/or light chain is identical or homologous to a certain type, type or subtype of antibody, and the remaining part is identical to another type of antibody.
  • a sort of, Antibodies of another class or subclass are the same or homologous as long as they have the desired biological activity (see, eg, US 4,816,567; and Morrison et al., 1984, PNAS, 81:6851-6855).
  • Chimeric antibodies useful in the present disclosure include primatized antibodies comprising variable region antigen-binding sequences and human constant region sequences from non-human primates (eg, monkeys, orangutans, etc.).
  • a "humanized" form of a non-human (eg, murine) antibody refers to a chimeric antibody that contains a minimal amount of non-human immunoglobulin sequence.
  • Most humanized antibodies are human recipient immunoglobulins whose hypervariable region residues have been replaced with non-human ones (e.g., mouse, rat, rabbit, or non-human primate) that have the desired specificity, affinity, and function. Hypervariable region residues (donor antibody).
  • donor antibody In some embodiments, framework region (FR) residues of human immunoglobulins are also replaced with non-human residues.
  • humanized antibodies may also contain residues that are not found in either the recipient antibody or the donor antibody. These modifications are intended to further optimize the antibody's performance.
  • Humanized antibodies generally contain at least one, and usually two, variable regions, in which all or nearly all hypervanable loops correspond to those of non-human immunoglobulins, while the FRs are entirely or almost entirely human immunoglobulins. Protein sequence.
  • the humanized antibody may also comprise at least a portion of an immunoglobulin constant region (Fc, typically a human immunoglobulin Fc).
  • Fc immunoglobulin constant region
  • Intact antibodies can be divided into different "classes” based on the amino acid sequence of the heavy chain constant region.
  • the five main classes are IgA, IgD, IgE, IgG and IgM, several of which can also be divided into different "subclasses” (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2.
  • the heavy chain constant regions of different classes of antibodies are called ⁇ , ⁇ , ⁇ , ⁇ , and ⁇ .
  • the subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known in the art.
  • Monoclonal antibodies used in the present disclosure can be produced by a number of methods.
  • monoclonal antibodies useful in the present disclosure can be obtained by hybridoma methods using cells from a number of species, including mouse, hamster, rat, and human (see, e.g., Kohler et al., 1975, Nature, 256:495). Either prepared by recombinant DNA technology (see, e.g., US 4,816,567), or isolated from a phage antibody library (see, e.g., Clackson et al., 1991, Nature, 352:624-628; and Marks et al., 1991, Journal of Molecular Biology , 222:581-597).
  • DAR drug to antibody ratio
  • the DAR of an ADC can range from 1 to 20, but depending on the number of attachment sites on the antibody, higher loadings are also possible.
  • the term DAR may be used when referring to the amount of drug loaded onto a single antibody, or alternatively, when referring to the average or mean DAR of a group of ADCs. DAR can also be calculated as the average DAR of the population of molecules in the product, i.e.
  • the average DAR value of the antibody drug conjugate of the invention is 1.0-20.0, such as 1.0-18.0, 1.0-16.0, 2.0-14.0, 3.0-12.0, 4.0-10.0, 5.0-9.0, 6.0- 8.0, 1.0-8.0, 2.0-6.0, such as 1.0, 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 ,3,3.1,3.2,3.3,3.4,3.5,3.6,3.7,3.8,3.9,4,4.1,4.2,4.3,4.4,4.5,4.6,4.7,4.8,4.9,5.0,5.1,5.2,5.3,5.4 ,5.5,5.6,5.7,5.8,5.9,6.0,6.1,6.2,6.3,6.4,6.5,6.6,6.7,6.8,6.9,7.0,7.1,7.2,7.3,7.4,7.5,7.6,7.7,7.8.0 ,7.9,8,8.1,8.2,8.3,8.4,
  • the term “comprises” or “includes” means the inclusion of the stated element, integer or step, but not the exclusion of any other element, integer or step.
  • combinations of the stated elements, integers, or steps are also encompassed unless otherwise specified.
  • reference is made to an antibody variable region that "comprises” a particular sequence it is also intended to encompass antibody variable regions that consist of that particular sequence.
  • the reagents and raw materials used in this disclosure are all commercially available.
  • the antibody-conjugated drugs of the protein-degrading bioactive compounds disclosed in the present disclosure can be used for the treatment of various hematological tumors and solid tumors, and have better therapeutic effects and/or better therapeutic effects than the corresponding protein-degrading bioactive compounds. Low side effects;
  • the conjugate of the present invention has better solubility and excellent chemical stability, and has a high drug-antibody ratio
  • the linker in the conjugate of the present invention has high plasma stability, but at the same time can be cleaved in the tumor microenvironment (both within tumor cells and outside tumor cells), so it can be used in tumors with low antigen expression or no antigen expression. produce good anti-tumor effects;
  • the conjugate (including ADC) of the present invention has better tumor tissue targeting, that is, the ability to enrich in the tumor microenvironment, increases the concentration ratio of bioactive molecules in the tumor and blood, and reduces the concentration ratio of the conjugate.
  • Mechanism-related toxicities with higher therapeutic index
  • the conjugate of the present invention has high stability in body circulation, reduces the shedding of drug molecules in non-target tissues, and reduces the "off-target” toxicity caused by the shedding of toxins in non-target tissues;
  • the bioactive molecules of the conjugate have higher anti-tumor cell activity and therefore have excellent by-stander effect.
  • the conjugate can more effectively kill tumor cells with high antigen expression and tumors. Tumor cells with low or no antigen expression in tissues; and/or
  • the disclosed drug-linker conjugate utilizes the extracellular cleavage ability of its linker in the tumor microenvironment to form an antibody-conjugated drug with an antibody without cell endocytosis ability. This type of antibody-conjugated drug still has high Antitumor activity.
  • Figure 1 Shows the degradation ability of compound D-2 and compound D-4 on c-Myc and GSPT1 proteins in HL60 cells.
  • Figure 2 shows that the ADC of the present invention can significantly degrade c-Myc protein in 22RV1 cells.
  • the measuring instrument of nuclear magnetic resonance uses a Bruker 400MHz nuclear magnetic resonance instrument; the measuring solvent is deuterated methanol (CD 3 OD), deuterated chloroform (CDCl 3 ) or hexadeuterated dimethyl sulfoxide (DMSO-d 6 ); the internal standard substance is tetramethylsilane (TMS).
  • NMR nuclear magnetic resonance
  • MS mass spectrometry
  • ESI Agilent 6120B
  • D-4-B is obtained by referring to the synthesis method of D-4.
  • Example 1.4-C 2-(2,6-dioxopiperidin-3-yl)-N-(4-((2-mercapto-N-(2-(naphthyl-1-yl)ethyl)) Synthesis of acetamido)methyl)benzyl)-1-oxoisoindoline-5-carboxamide (D-4-C)
  • D-4-C is prepared from D-4-C-1.
  • the specific reaction equation and operation steps are as follows:
  • Example 1.7-A 4-((4-(3-chloro-4-(hydroxymethyl)phenyl)piperazin-1-yl)methyl)-N-((2-(2,6-di Oxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl (D-7-A)
  • D-7-A-2 is synthesized as follows:
  • Step 2 and Step 3 For the synthesis operation of D-8-2 and D-8-3, please refer to D-7-4 and D-7
  • Example 1.9-A 4-((4-(3-chloro-4-methylphenyl)piperazin-1-yl)methyl)-N-((2-(2,6-dioxopiperine) (Din-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(2-hydroxyethoxy)benzamide (D-9-A)
  • D-9-A-1 instead of D-9-4, refer to the preparation method of D-9 to obtain D-9-A
  • D-9-A-1 is prepared as follows:
  • Example 1.9-B 4-((4-(3-chloro-4-methylphenyl)-2-(hydroxymethyl)piperazin-1-yl)methyl)-N-((2-( 2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)benzamide (D-9-B)
  • D-9-B-1 instead of D-9-2, refer to the preparation method of D-9 to obtain D-9-B.
  • D-9-B-1 is synthesized as follows:
  • D-11-4 and D-11 refer to the preparation methods of D-7-4 and D-7 respectively, and finally D-11 is obtained.
  • step 2 refers to the preparation of D-8, and D-12 is obtained.
  • D-13-A was prepared by referring to the method of D-13.
  • D-13-B-2 was prepared by referring to the D-13 method.
  • D-14 is prepared from D-13 by reduction.
  • the specific method is as follows:
  • D-15-3 and D-15-4 For the synthesis operations of D-15-3 and D-15-4, refer to the preparation methods of D-7-4 and D-7 respectively, and then use the obtained D-15-4 as raw material to prepare D-15 according to the following method:
  • D-16-2, D-16-3, D-16-4 and D-16-5 refer to D-13-1, D-13-A-1, D-7-4 and D-7 respectively. , obtain D-16-5; then use D-16-5 as raw material and synthesize D-16 by referring to the preparation method of D-8.
  • D-17-4 refers to the preparation method of D-15-2.
  • D-17-5 and D-17-6 refer to the preparation methods of D-7-3 and D-7 respectively to obtain intermediate D-17- 6; Then use D-17-6 as raw material and refer to step 5 corresponding to D-15 to obtain D-17.
  • Example 2.1 N-((10S,13S)-10-(4-(dipropylamino)butyl)-1,1,1-trifluoro-14-methyl-6,9,12-trioxo -3-oxa-5,8,11-triazapentadecan-13-yl)-6-(2-(methanesulfonyl)pyrimidin-5-yl)hex-5-ynamide (L-1 )Synthesis
  • step 2 To the DMF solution of L-1-3 obtained in step 2, add L-1-4 (7.0 g, 12.7 mmol) and DMF (35 mL). Cool the reaction solution to -15°C, add DMTMM (4.2g, 15.2mmol), -10°C The reaction was stirred for 2 h. Pour the reaction solution into DCM (200 mL), and wash the organic phase with 2% NaHCO 3 aqueous solution (100 mL*3), water (100 mL) and saturated NaCl aqueous solution (50 mL).
  • Example 2.2 N 6 ,N 6 -dipropyl-N 2 -((6-(2-(methanesulfonyl)pyrimidin-5-yl)hex-5-ynyl)-L-valine)- Synthesis of L-lysine (L-2)
  • Example 3.1 2-(2,6-dioxopiperidin-3-yl)-N-(4-((12S,15S)-12-(4-(dipropylamino)butyl)-15 -Isopropyl-22-(2-(methylsulfonyl)pyrimidin-5-yl)-2-(2-(naphthyl-1-yl)ethyl)-3,8,11,14,17-penta Oxo-5-oxa-2,7,10,13,16-pentaazabis-21-yn-1-yl)benzyl)-1-oxoisoindoline-5-methyl Synthesis of Amide (DL-1)
  • VH amino acid sequence SEQ ID NO:1 Connect the VH amino acid sequence SEQ ID NO:1 to the heavy chain constant region amino acid sequence of IgG1 (SEQ ID NO:2), and then construct it into pcDNA3.4 through codon optimization and gene synthesis (Shanghai Baiying Biotechnology Co., Ltd.) On the carrier, it is named B21231602H.
  • VL amino acid sequence SEQ ID NO:3 Connect the VL amino acid sequence SEQ ID NO:3 to the light chain constant region amino acid sequence of IgG1 (SEQ ID NO:4), and then construct it into pcDNA3.4 through codon optimization and gene synthesis (Shanghai Baiying Biotechnology Co., Ltd.) On the carrier, it is named B21231602L.
  • PSMA mAb shows heavy chain amino acid sequence
  • light chain amino acid sequence is SEQ ID NO: 6
  • the purpose of this experiment is to study the affinity of PSMA mAb (prepared in Example 4.1) to human PSMA protein and provide a basis for PSMA mAb in vitro and in vivo efficacy testing.
  • the negative control is human IgG1 with native sequence.
  • ELISA coating buffer carbonate buffer, pH 9.5, Biolegend, 421701
  • dilute human PSMA protein purchased from: Acrobiosystems product number, PSA-H52H3
  • ELISA wash buffer purchased from: Multiscience, product number, EK0011
  • 300ul/well wash three times, and pat the plate dry.
  • Add 100ul/well of ELISA assay dilute buffer purchased from: biolegend Cat. No. 421203 to the enzyme plate, and incubate at 37°C for 1 hour. Clean again.
  • the results show that PSMA mAb has good binding activity to human PSMA protein, and the negative control human IgG1 does not bind to human PSMA.
  • the EC50 was calculated based on binding curve fitting.
  • the EC50 value of PSMA mAb for human PSMA protein was 6.67nM.
  • the purpose of this experiment is to study the endocytic activity of PSMA mAb (Progenics) in 22RV cells and provide a basis for the in vivo and in vitro efficacy testing of PSMA mAb (Progenics).
  • the PSMA antibody showed an increasing internalization rate as the incubation time prolonged, that is, the endocytic activity increased with the prolongation of the incubation time, and finally reached a saturation state.
  • the internalization rate % was calculated, that is, when the PSMA antibody was incubated with 22RV1 cells for 18 hours, the internalization rate was 60%.
  • PSMA mAb (8.84mg/mL) obtained in Example 4.1, dilute it with 0.005mL of 20mM PB+100mM disodium edetate solution (pH 7.6), adjust the pH to 7.45 with 0.5M Na 2 HPO 4 solution, and add Mix 20mM TCEP (tris(2-carboxyethyl)phosphine, 0.0082ml, 0.164 ⁇ mol) solution and leave it at room temperature for 90 minutes. Add a solution of DL-1 (0.470 mg, 12 times the molar amount of antibody material) in dimethyl sulfoxide (0.0558 mL) to the above solution, mix well, and let stand at room temperature for 2 hours.
  • DL-1 (0.470 mg, 12 times the molar amount of antibody material
  • the method for measuring DAR value is as follows:
  • Example 5.1 Using a similar operation to Example 5.1, replacing DL-1 with DL-2, ADC-002, the coupling product of DL-2 and PSMA mAb, was obtained.
  • the DAR value determined by mass spectrometry was 8.0.
  • LC represents the antibody light chain
  • HC represents the antibody heavy chain
  • DAR1 represents the conjugate containing a light chain or heavy chain coupled to a toxin linker
  • DAR2 represents a conjugate containing a light chain or heavy chain coupled to two toxin linkers.
  • Conjugates DAR3 represents conjugates containing three toxin linkers coupled to a light or heavy chain.
  • LC, HC, DAR1, DAR2, and DAR3 are as described above.
  • Example 6.1 Test of protein degradation activity of protein-degrading bioactive compounds on c-Myc and GSPT1 proteins
  • Cell culture conditions The cells in the examples of the present invention were all purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences. Among them, human acute promyelocytic leukemia cells (HL60 cells) were cultured in RPMI-1640 (meilunbio, MA0215) + 20% FBS (manufacturer: BI, product number: 04-001-1ACS) + 1% penicillin/streptomycin (manufacturer: Hyclone, Cat. No.: SH40003.01) culture medium. Cell culture conditions were all 5% CO2 concentration and 37% humidity. When the coverage rate reaches about 80%, the cells are passaged in a ratio of 1:3.
  • the HL60 cells are in the exponential growth phase, the HL60 cells are plated into a 6-well plate with 1 ⁇ 10 6 cells per well.
  • the compound of the present invention is added to culture the cells, and the protein is extracted after 6 hours of drug treatment.
  • BCA protein concentration determination kit from Thermo Fisher, Cat. No. 23225
  • BSA standard determination solution obtained in the above step 2.2 for the Western blotting (WB) experiment according to Table 1 below.
  • WB Western blotting
  • Table 1 below. (can be diluted and tested), use a 96-well plate to add samples, add PBS to each well to 20 ⁇ l, add 200 ⁇ l BCA working solution (prepared according to the kit), mix well and place it at 60°C for 10 minutes, then detect the absorbance at 562nm. After recording the readings, use the standard concentration gradient to make a standard curve, and substitute the sample absorbance to calculate the sample protein concentration.
  • PVDF membrane (Millipore, Cat. No. ISEQ00010) by wet transfer method (PVDF membrane needs to be activated with methanol for 1 minute before use), 300mA, 2h, during the transfer process Generates a lot of heat and requires an ice box to cool down.
  • the protein degradation ability in the above cells was tested respectively, and the results are shown in Figure 1.
  • the Blank group represents the DMSO blank group without adding the test compound.
  • Figure 1 shows that compound D-2 and compound D-4 can completely degrade c-Myc and GSPT1 proteins in HL60 cells at lower concentrations (3-10 nM).
  • Example 6.2 Detection of inhibitory activity of protein-degrading bioactive compounds on HL60 cells
  • HL60 cells Count the HL60 cells in the logarithmic growth phase and spread them evenly in a 96-well transparent bottom white plate.
  • the number of cells is 20,000 per well and 100 ⁇ L per well.
  • test results showed that all the tested compounds had potent proliferation inhibitory activity on HL60 cells.
  • Example 6.3 Detection of inhibitory activity of compounds or ADC on 22RV1 cells in vitro
  • Count the 22RV1 Cell Bank of the Chinese Academy of Sciences, TCHu100 cells in the logarithmic growth phase and spread them evenly in a 96-well transparent bottom white plate.
  • the number of cells is 10,000-20,000 per well, 100 ⁇ L per well.
  • test results show that the ADC molecules disclosed in the present invention have tumor cell killing effect, and compared with the small molecules themselves, the anti-proliferative activity after being prepared into ADC is significantly improved.
  • Example 6.4 ADC drug degradation test on c-Myc protein in 22RV1 cells
  • Cell culture conditions The cells in this example were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences. Among them, human prostate cancer cells (22RV-1 cells) were cultured in RPMI-1640 (meilunbio, MA0215) + 10% FBS (manufacturer: BI, product number: 04-001-1ACS) + 1% penicillin/streptomycin (manufacturer: Hyclone , Catalog No.: SH40003.01) in culture medium. Cell culture conditions were all 5% CO2 concentration and 37% humidity. When the coverage rate reaches about 80%, the cells are passaged in a ratio of 1:3.
  • the 22RV-1 cells are in the exponential growth phase, the 22RV-1 cells are plated into a 6-well plate, with 1 ⁇ 10 6 cells per well.
  • the ADC drug of the present invention is added to culture the cells, and the protein is extracted after 6 hours of drug treatment.
  • BCA protein concentration determination kit from Thermo Fisher, Cat. No. 23225
  • BSA standard determination solution and the supernatant for Western blotting (WB) experiment obtained in step 2.2 according to Table 1 in Example 6.1 (upper The clear solution can be diluted and tested)
  • WB Western blotting
  • the clear solution can be diluted and tested
  • use a 96-well plate to add samples, make up each well to 20 ⁇ l with PBS, then add 200 ⁇ l BCA working solution (prepared according to the kit), mix well and place it at 60°C for 10 minutes before detecting at 562nm.
  • Absorbance, after recording the reading use the standard concentration gradient to make a standard curve, and substitute it into the sample absorbance to calculate the sample protein concentration.
  • PVDF membrane (Millipore, Cat. No. ISEQ00010) by wet transfer method (PVDF membrane needs to be activated with methanol for 1 minute before use), 300mA, 2h, during the transfer process Generates a lot of heat and requires an ice box to cool down.
  • test results are shown in Figure 2 (where Ctrl refers to the use of solvent instead of drug in step 2.1 cell treatment, that is, the solvent control group).
  • the results show that the prepared ADC can significantly degrade the c-Myc protein in 22RV1 cells.
  • Human peripheral blood mononuclear cells hPBMCs in the logarithmic growth phase (purchased from Shanghai Aoneng Biotechnology Co., Ltd., the culture medium is RPMI1640 (Meilun Bio, MA 0215), containing 10% FBS (gibco, 16000-044) and 1 % penicillin/streptomycin (Hyclone, SV30010); human prostate cancer cell Vcap (purchased from ATCC, the culture medium is DMEM (Hyclone, product number: SH30022.01) + 10% FBS (manufacturer: BI, product number: 04-001- 1ACS) + 1% penicillin/streptomycin (Manufacturer: Hyclone, Catalog No.: SH40003.01), count the cells, and spread them evenly in a 96-well transparent bottom white plate. The number of cells is 5,000 to 10,000 per well, 100 ⁇ L per well.
  • ADC has reduced inhibitory activity on PSMA-negative HL60 cells and normal human cell hPBMC cells. After being prepared into ADC, the therapeutic window is greatly improved.
  • FLOH1 PSMA gene name
  • the expression levels of FLOH1 in different cell lines can be found in The Human Protein Atlas (https://www.proteinatlas.org/). The data shows that FLOH1 is not expressed in HL60 cells; Vcap and 22RV1 express FLOH1. Detailed results See Table 7.
  • mice NOD/SCID mice, male, 4-6 weeks old, purchased from Vitong Lever Experimental Animal Co., Ltd.
  • 22RV1 cells are cultured in RPMI-1640 culture medium containing 10% fetal calf serum and passaged in about 3-4 days. After the cells grow to a sufficient number, adjust the cell concentration with a mixture of pre-cooled PBS and Matrigel (1:1). to approximately 2.5 ⁇ 10 7 cells/mL.
  • NOD/SCID mice were adapted to the laboratory environment for 3-5 days. 22RV1 cells were subcutaneously inoculated on the right front back at a volume of 5 ⁇ 10 6 cells/mouse and an inoculation volume of 0.2 ml (containing 50% Matrigel). Wait until the tumor grows to When the diameter is about 100 ⁇ 150mm 3 , they are randomly divided into groups and experiments are carried out.
  • the tumor-bearing nude mice enrolled in Table 8 were administered according to the following regimen.
  • Vehicle indicates that the vehicle replaced the drug as a control group.
  • TGI% (1-T/C) * 100% (T and C are the values of the treatment group and the control group, respectively.
  • V Relative tumor volume
  • ADC-002 has a significant anti-tumor growth effect on the 22RV1 transplanted tumor model.
  • D21 the body weight of all tested dose groups decreased slightly during the administration, and there was no obvious drug toxicity.

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Abstract

An antibody-drug conjugate containing an Myc protein degradation agent bioactive compound, a preparation method therefor, and use thereof in preventing and/or treating diseases related to abnormal cell activity, including but not limited to use in preventing and/or treating neoplastic diseases.

Description

包含Myc蛋白降解剂类生物活性化合物的抗体药物偶联物及其制备方法和用途Antibody drug conjugates containing Myc protein degrader bioactive compounds and preparation methods and uses thereof
本发明要求2022年8月4日提交的中国专利申请CN202210932118.2和2022年12月20日提交的中国专利申请CN202211640683.8的优先权，该优先权文件的说明书、说明书附图和权利要求书所记载的内容全文引入本发明的说明书并被作为本发明说明书原始记载的一部分。申请人进一步声明，申请人拥有基于该优先权文件修改本发明的说明书和权利要求书的权利。The present invention claims the priority of Chinese patent application CN202210932118.2 submitted on August 4, 2022 and Chinese patent application CN202211640683.8 submitted on December 20, 2022, the description, description drawings and claims of the priority document The entire contents described are incorporated into the specification of the present invention and shall be regarded as part of the original description of the specification of the present invention. The applicant further declares that the applicant has the right to amend the description and claims of the invention based on the priority document.
技术领域Technical field
本公开属于医药技术领域，涉及蛋白降解剂类生物活性化合物的抗体药物偶联物、蛋白降解剂类生物活性化合物、药物连接体偶联物及其制备方法，以及其在预防和/或治疗与细胞活动异常相关的疾病，包括但不限于在预防和/或治疗肿瘤疾病中的用途。The present disclosure belongs to the field of medical technology and relates to antibody-drug conjugates of protein-degrading bioactive compounds, protein-degrading bioactive compounds, drug linker conjugates and their preparation methods, as well as their use in prevention and/or treatment. Diseases related to abnormal cell activity, including but not limited to use in the prevention and/or treatment of tumor diseases.
背景技术Background technique
很多疾病都与细胞内蛋白质功能异常有关，治疗这些疾病的主要手段是小分子化合物。然而，80％以上蛋白质由于缺乏可产生能够成药作用的位点，这类靶点被认为是不能通过传统小分子成药的。Many diseases are related to intracellular protein dysfunction, and the main means of treating these diseases are small molecule compounds. However, more than 80% of proteins lack sites that can produce druggable effects. Such targets are considered to be undruggable through traditional small molecules.
靶向蛋白降解剂(Targeted Protein Degradation，TPD)，如蛋白降解靶向嵌合体(PROTAC)和分子胶降解剂(MGD)，均通过泛素-蛋白酶体途径介导靶蛋白降解，不需要与影响蛋白活性的位点紧密结合就可以发挥作用，使得上述“不可成药”的蛋白质能够成为新的药物靶点。同时，靶向蛋白降解疗法可以持续诱导致病蛋白快速、高效降解，降低了靶点蛋白耐药性的产生。Targeted Protein Degradation (TPD), such as protein degradation targeting chimera (PROTAC) and molecular glue degrader (MGD), all mediate target protein degradation through the ubiquitin-proteasome pathway without affecting Protein active sites can function by tightly binding, making the above-mentioned "undruggable" proteins a new drug target. At the same time, targeted protein degradation therapy can continuously induce rapid and efficient degradation of disease-causing proteins, reducing the development of drug resistance to target proteins.
Myc，是一个重要的转录因子家族，包括c-Myc、n-Myc、l-Myc，它们对于细胞生长，代谢和组织发育至关重要。同时，Myc也是一组重要的癌基因，c-Myc在约30-50％的人类恶性肿瘤中异常表达，在影响细胞的生长、增殖、分化、凋亡以及细胞的代谢和恶性转化中均发挥着极为重要的作用。c-Myc的异常高表达与许多人类癌症的不良预后有关。但由于c-Myc特殊的蛋白结构，直接靶向抑 制c-Myc的疗法抑制未能成功，而且，靶向c-Myc转录机制的方法也成效甚微。因此，c-Myc长期以来被认为是不可成药靶点。Myc is an important family of transcription factors, including c-Myc, n-Myc, and l-Myc, which are essential for cell growth, metabolism and tissue development. At the same time, Myc is also an important group of oncogenes. c-Myc is abnormally expressed in about 30-50% of human malignant tumors and plays a role in affecting cell growth, proliferation, differentiation, apoptosis, as well as cell metabolism and malignant transformation. plays an extremely important role. Abnormally high expression of c-Myc is associated with poor prognosis in many human cancers. However, due to the special protein structure of c-Myc, it directly targets inhibitors Therapeutic inhibition of c-Myc has been unsuccessful, and approaches targeting the c-Myc transcriptional machinery have had limited success. Therefore, c-Myc has long been considered an undruggable target.
靶向蛋白降解剂和分子胶降解剂的最大优势之一就是能够使靶点从“不可成药”变为“可成药”，但由于其存在成药性质不佳(例如口服生物利用度低)、脱靶导致的剂量限制性毒性等问题，使得该类分子的开发受到较大限制。One of the biggest advantages of targeted protein degraders and molecular glue degraders is that they can change the target from "undruggable" to "druggable". However, due to their poor drug properties (such as low oral bioavailability), off-target The resulting dose-limiting toxicity and other problems have greatly restricted the development of this type of molecules.
发明内容Contents of the invention
本发明涉及抗体偶联药物(ADC)及其用途。ADC药物融合了抗体的肿瘤靶向作用和生物活性化合物的高活性，成为一种生物导弹，具有非常可期待的疗效和安全性优势。小分子化合物与抗体偶联形成ADC后，溶解度大幅提高，可实现静脉给药，有效解决小分子化合物生物利用度等成药性问题。抗体引导ADC结合到靶细胞，实现肿瘤组织富集作用，减少非靶组织暴露，降低生物活性化合物通过系统给药可能带来的毒性。和肿瘤细胞结合的ADC可以在肿瘤微环境中裂解释放生物活性分子，杀伤肿瘤细胞。ADC也可以被肿瘤细胞内化，在细胞内在特定酶作用下发生酶解释放小分子药物，治疗疾病。因此可以期待由蛋白降解剂和肿瘤靶向抗体联合组成的ADC可以实现肿瘤富集，消除或降低蛋白降解剂作用于非疾病组织而引起的毒副作用，提高治疗效果。利用ADC技术实现蛋白降解剂的肿瘤靶向性，降低其毒副作用，将具有很高的临床价值。The present invention relates to antibody drug conjugates (ADCs) and their uses. ADC drugs combine the tumor targeting effect of antibodies and the high activity of biologically active compounds, becoming a biological missile with very promising efficacy and safety advantages. After conjugating small molecule compounds with antibodies to form ADCs, the solubility is greatly improved, enabling intravenous administration and effectively solving pharmaceutical problems such as the bioavailability of small molecule compounds. Antibodies guide ADCs to bind to target cells, achieving tumor tissue enrichment, reducing non-target tissue exposure, and reducing possible toxicity caused by systemic administration of bioactive compounds. ADCs that bind to tumor cells can cleave and release bioactive molecules in the tumor microenvironment to kill tumor cells. ADC can also be internalized by tumor cells and undergo enzymatic decomposition under the action of specific enzymes in the cells to release small molecule drugs to treat diseases. Therefore, it is expected that an ADC composed of a protein degrading agent and a tumor-targeting antibody can achieve tumor enrichment, eliminate or reduce the toxic side effects caused by the protein degrading agent acting on non-diseased tissues, and improve the therapeutic effect. Using ADC technology to achieve tumor targeting of protein degradation agents and reduce their toxic and side effects will have high clinical value.
为此，在本公开的第一方面，本公开提供式I所示的抗体药物偶联物，
To this end, in a first aspect of the present disclosure, the present disclosure provides an antibody drug conjugate represented by Formula I,
或所述抗体药物偶联物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物，or a stereoisomer of the antibody drug conjugate, a prodrug thereof, a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable solvate thereof,
其中，in,
Tb为抗体或其抗原结合片段；Tb is an antibody or its antigen-binding fragment;
S为Tb上的硫原子；S is the sulfur atom on Tb;
L为连接体，其共价结合Tb和D；L is the linker, which covalently binds Tb and D;
D为Myc蛋白降解剂片段；D is the Myc protein degrader fragment;
q为选自1-20的整数。 q is an integer selected from 1-20.
基于上下文，应当理解的是，“L为连接体，其共价结合Tb和D”意指连接体L与Tb上的S原子共价结合且与D共价结合。下文中的类似表述具有类似的含义。Based on the context, it should be understood that "L is a linker that covalently binds Tb and D" means that the linker L is covalently bound to the S atom on Tb and is covalently bound to D. Similar expressions below have similar meanings.
在一些实施方案中，提供式II所示的抗体药物偶联物，
In some embodiments, an antibody drug conjugate represented by Formula II is provided,
其中，in,
L1为接头单元，其共价结合抗体或其抗原结合片段(Tb)与L2；L 1 is a linker unit, which covalently binds the antibody or its antigen-binding fragment (Tb) to L 2 ;
L2为连接单元，其共价结合接头单元(L1)与L3；L 2 is a connecting unit, which covalently binds the linker unit (L 1 ) and L 3 ;
L3选自氨基酸残基或由2-10个(例如2、3、4、5、6、7、8、9或10个)氨基酸残基组成的短肽；所述氨基酸残基选自天然氨基酸残基、非天然氨基酸残基或AA1所示氨基酸残基或其立体异构体；L 3 is selected from amino acid residues or a short peptide consisting of 2-10 (such as 2, 3, 4, 5, 6, 7, 8, 9 or 10) amino acid residues; the amino acid residues are selected from natural Amino acid residues, unnatural amino acid residues or amino acid residues shown in AA 1 or their stereoisomers;
AA1所示氨基酸残基的结构如下所示：
The structure of the amino acid residue shown in AA 1 is as follows:
Rx、Ry任一个为氢，另一个选自或者，Rx与Ry和与它们共同相连的碳原子一起，形成4-10元杂环，所述4-10元杂环任选地被一个或多个R0所取代；Either R x or R y is hydrogen, and the other is selected from Alternatively, R x and R y and the carbon atoms to which they are commonly connected form a 4-10 membered heterocyclic ring, which is optionally substituted by one or more R 0 ;
Rx1、Ry1各自独立地选自氢、C1-6烷基；或者，Rx1与Ry1和与它们共同相连的氮原子一起，形成4-10元杂环，所述4-10元杂环任选地被一个或多个R0’所取代；R x1 and R y1 are each independently selected from hydrogen and C 1-6 alkyl; alternatively, R x1 and R y1 and the nitrogen atoms commonly connected to them form a 4-10 membered heterocyclic ring, and the 4-10 membered heterocycle The heterocycle is optionally substituted with one or more R 0' ;
R0、R0’各自独立地选自C1-6烷基、C3-6环烷基、-NRx2Ry2和任选被C1-6烷基取代的4-10元杂环基；R 0 and R 0' are each independently selected from C 1-6 alkyl, C 3-6 cycloalkyl, -NR x2 R y2 and 4-10 membered heterocyclyl optionally substituted by C 1-6 alkyl. ;
Rx2、Ry2各自独立地选自氢和C1-6烷基；R x2 and R y2 are each independently selected from hydrogen and C 1-6 alkyl;
L4为化学键或间隔单元，共价结合D与L3；L 4 is a chemical bond or spacer unit, covalently combining D and L 3 ;
Tb、q和D的定义如本公开中任一方案所述。 Tb, q and D are as defined in any aspect of this disclosure.
在一些实施方案中，所述q选自1-18、1-16、1-14、1-12、1-10、1-8、2-8、或4-6；例如为选自1-10的整数。In some embodiments, the q is selected from 1-18, 1-16, 1-14, 1-12, 1-10, 1-8, 2-8, or 4-6; for example, is selected from 1-1 An integer of 10.
优选地，所述q选自1、2、3、4、5、6、7、8、9和10。Preferably, said q is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10.
优选地，所述q选自2、4、6和8。Preferably, said q is selected from 2, 4, 6 and 8.
在一些实施方案中，L1选自 1位通过S原子与Tb相连，2位与L2相连。In some embodiments, L is selected from Position 1 is connected to Tb through the S atom, and position 2 is connected to L 2 .
在一些实施方案中，L1选自1位通过S原子与Tb相连，2位与L2相连。In some embodiments, L is selected from Position 1 is connected to Tb through the S atom, and position 2 is connected to L 2 .
在一些实施方案中，L2选自1位与L1相连，2位与L3相连，In some embodiments, L2 is selected from Bit 1 is connected to L 1 , bit 2 is connected to L 3 ,
W选自-CH2-、-OCH2CH2-，W is selected from -CH 2 -, -OCH 2 CH 2 -,
W’选自-CRx3Ry3-和-NRx3-，W' is selected from -CR x3 R y3 - and -NR x3 -,
Rx3、Ry3各自独立地选自氢、C1-4烷基、C2-4烯基、C2-4炔基、C3-6环烷基、3-6元杂环基，R x3 and R y3 are each independently selected from hydrogen, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 3-6 cycloalkyl, and 3-6 membered heterocyclyl,
或者，Rx3与Ry3和与它们共同相连的碳原子一起，形成3-6元碳环或3-6元杂环，Alternatively, R x3 and R y3 together with the carbon atoms connected to them form a 3-6 membered carbocyclic ring or a 3-6 membered heterocyclic ring,
p为选自0-10之间的任意整数。p is any integer selected from 0-10.
优选地，W为-CH2-。Preferably, W is -CH2- .
优选地，W’选自-CH2-、-C(CH3)2-、-N(CH3)-和-NH-。Preferably, W' is selected from -CH 2 -, -C(CH 3 ) 2 -, -N(CH 3 )- and -NH-.
优选地，Rx3、Ry3各自独立地选自氢、甲基。Preferably, R x3 and R y3 are each independently selected from hydrogen and methyl.
优选地，p选自0、1、2、3和4。Preferably, p is selected from 0, 1, 2, 3 and 4.
在一些实施方案中，L2选自 1位和L1相连，2位和L3相连。 In some embodiments, L2 is selected from Bit 1 is connected to L 1 , bit 2 is connected to L 3 .
在一些实施方案中，L1-L2选自 1位通过S原子与Tb相连，2位与L3相连，W、W’和p的定义如本公开中任一方案所述。In some embodiments, L 1 -L 2 are selected from Position 1 is connected to Tb through an S atom, position 2 is connected to L3 , and W, W' and p are defined as described in any scheme of this disclosure.
在一些实施方案中，L1-L2选自In some embodiments, L 1 -L 2 are selected from
1位通过S原子与Tb相连，2位与L3相连。 Position 1 is connected to Tb through the S atom, and position 2 is connected to L 3 .
在一些实施方案中，L1-L2选自
1位通过S原子与Tb相连，2位与L3相连。In some embodiments, L 1 -L 2 are selected from
Position 1 is connected to Tb through the S atom, and position 2 is connected to L 3 .
在一些实施方案中，L3选自AA1所示氨基酸残基或包含AA1所示氨基酸残基的2-10个(例如2、3、4、5、6、7、8、9或10个)氨基酸残基组成的短肽；所述氨基酸残基选自天然氨基酸残基、非天然氨基酸残基或AA1所示氨基酸残基或其立体异构体。In some embodiments, L 3 is selected from the amino acid residues represented by AA 1 or includes 2-10 of the amino acid residues represented by AA 1 (e.g., 2, 3, 4, 5, 6, 7, 8, 9 or 10 ) A short peptide composed of amino acid residues; the amino acid residues are selected from natural amino acid residues, non-natural amino acid residues or amino acid residues represented by AA 1 or their stereoisomers.
在一些实施方案中，L3选自Val-Cit、Val-Ala、Phe-Lys、Val-Lys、Val-AA1、Ala-Ala-Ala、Ala-Ala-Asn、Val-AA1-Gly、Gly-Gly-Phe-Gly。In some embodiments, L3 is selected from Val-Cit, Val-Ala, Phe-Lys, Val-Lys, Val- AA1 , Ala-Ala-Ala, Ala-Ala-Asn, Val- AA1 -Gly, Gly-Gly-Phe-Gly.
在一些实施方案中，L3选自Val-AA1-Gly。In some embodiments, L3 is selected from Val- AA1 -Gly.
在一些实施方案中，AA1所示氨基酸残基的结构如下所示：
In some embodiments, the structure of the amino acid residue represented by AA 1 is as follows:
Rx、Ry任一个为氢，另一个选自Rx1、Ry1各自独立地选自氢、C1-6烷基。 Either R x or R y is hydrogen, and the other is selected from R x1 and R y1 are each independently selected from hydrogen and C1-6 alkyl.
优选地，Rx1、Ry1各自独立地选自甲基、乙基、正丙基和正丁基；优选地，Rx1、Ry1均为正丙基。Preferably, R x1 and R y1 are each independently selected from methyl, ethyl, n-propyl and n-butyl; preferably, R x1 and R y1 are both n-propyl.
在一些实施方案中，L4选自化学键、 1位与L3相连，2位与D相连。In some embodiments, L4 is selected from chemical bonds, Bit 1 is connected to L 3 and bit 2 is connected to D.
在一些实施方案中，L4为1位与L3相连，2位与D相连。In some embodiments, L 4 is Bit 1 is connected to L 3 and bit 2 is connected to D.
在一些实施方案中，D为Myc蛋白降解剂片段，例如c-Myc蛋白降解剂片段、n-Myc蛋白降解剂片段等。In some embodiments, D is a Myc protein degrading agent fragment, such as a c-Myc protein degrading agent fragment, an n-Myc protein degrading agent fragment, etc.
在一些实施方案中，D为c-Myc蛋白降解剂片段。In some embodiments, D is a c-Myc protein degrader fragment.
在一些实施方案中，D为式III所示的结构单元，D通过其含有的氧原子、硫原子或氮原子与L4相连；
In some embodiments, D is a structural unit represented by Formula III, and D is connected to L 4 through the oxygen atom, sulfur atom or nitrogen atom it contains;
其中，in,
R1选自：Ra(CH2)a-、Ra(CH2)aC(＝O)-、Ra(CH2)aNHC(＝O)-、Ra(CH2)aOC(＝O)-或Ra(CH2)aS(＝O)2-，其中“CH2”任选被一个或多个R9取代，或者任选被-CH2CH2-取代，其中“NH”任选被R14取代；R 1 is selected from: R a (CH 2 ) a -, R a (CH 2 ) a C (=O)-, R a (CH 2 ) a NHC (= O) -, R a (CH 2 ) a OC (=O)- or R a (CH 2 ) a S(=O) 2 -, where "CH 2 " is optionally substituted by one or more R 9 , or optionally substituted by -CH 2 CH 2 -, where "NH" is optionally substituted by R 14 ;
Q选自：-NR2-、-O-、 Q is selected from: -NR 2 -, -O-,
R2选自：氢、Rb-、RbC(＝O)-、RbS(＝O)2-、R14；R 2 is selected from: hydrogen, R b -, R b C(=O)-, R b S(=O) 2 -, R 14 ;
环A表示至少含有一个N原子作为杂原子的杂环烷基，其通过N原子与T或R1相连，环A任选被一个或多个选自R9的基团取代；Ring A represents a heterocycloalkyl group containing at least one N atom as a heteroatom, which is connected to T or R 1 through an N atom. Ring A is optionally substituted by one or more groups selected from R 9 ;
Ra、Rb选自：C1-C8烷基、C3-C10环烷基、C3-C10桥环基、-NR11R12、C3-C10任含O、S、SO2、N或NHC(＝O)R22的杂环基、芳基、杂芳基、稠合芳基环烷基、稠合芳基杂环基、稠合杂芳基环烷基、稠合杂芳基杂环基、芳基-烷基、杂芳基- 烷基、芳基-环烷基、芳基-杂环基、环烷基-杂环基、杂环基-杂环基，所述C1-C8烷基中的任意两个C-C之间可以插入选自-O-、-S-、-C(＝O)-、-S(＝O)-、-S(＝O)2-、-NH-、-NHC(＝O)-、-NHC(＝O)NH-、-NHS(＝O)2-中的一种的基团，Ra任选被一个或多个R9取代，Rb任选被一个或多个R10取代；R a and R b are selected from: C 1 -C 8 alkyl, C 3 -C 10 cycloalkyl, C 3 -C 10 bridged cyclic group, -NR 11 R 12 , C 3 -C 10 optionally containing O and S , SO 2 , N or NHC(=O)R 22 heterocyclyl, aryl, heteroaryl, fused arylcycloalkyl, fused arylheterocyclyl, fused heteroarylcycloalkyl, Fused heteroarylheterocyclyl, aryl-alkyl, heteroaryl- Alkyl, aryl-cycloalkyl, aryl-heterocyclyl, cycloalkyl-heterocyclyl, heterocyclyl-heterocyclyl, between any two CCs in the C 1 -C 8 alkyl group Can be inserted selected from -O-, -S-, -C(=O)-, -S(=O)-, -S(=O) 2 -, -NH-, -NHC(=O)-, - A group of one of NHC(=O)NH- and -NHS(=O) 2 -, R a is optionally substituted by one or more R 9 , R b is optionally substituted by one or more R 10 ;
T、U、Z分别独立地选自：化学键、N、O、羰基、C1-C6亚烷基、C3-C10亚环烷基、亚芳基、亚杂芳基、亚杂环基，所述亚烷基、亚环烷基、亚芳基、亚杂芳基、亚杂环基任选被一个或多个R9取代；T, U, and Z are each independently selected from: chemical bond, N, O, carbonyl group, C 1 -C 6 alkylene group, C 3 -C 10 cycloalkylene group, arylene group, heteroarylene group, heterocyclic ring Base, the alkylene, cycloalkylene, arylene, heteroarylene, heterocyclylene is optionally substituted by one or more R 9 ;
Y选自：化学键、-C(＝O)NH(CH2)b-、-NHC(＝O)(CH2)b-、-O(CH2)b-、-NR2(CH2)b-，其中“CH2”任选被一个或多个R9取代，或者任选被-CH2CH2-取代，其中“NH”任选被R14取代；Y is selected from: chemical bond, -C(=O)NH(CH 2 ) b -, -NHC(=O)(CH 2 ) b -, -O(CH 2 ) b -, -NR 2 (CH 2 ) b -, wherein "CH 2 " is optionally substituted by one or more R 9 , or optionally substituted by -CH 2 CH 2 -, wherein "NH" is optionally substituted by R 14 ;
X选自：-(CR7R8)o-、-C(＝O)-；X is selected from: -(CR 7 R 8 ) o -, -C(=O)-;
R3-R5、R7-R9分别独立地选自：氢、R13、卤素、氰基、氨基、羟基、巯基、硝基、-R21N(R22)R22、-R21C(＝O)R22、-R21NHC(＝O)R22、-R21C(＝O)NHR22、-R21C(＝O)OR22、-R21OC(＝O)R22、-R21S(＝O)2R22、-R21S(＝O)2NHR22、C1-C6烷基、C1-C6烷氧基、C1-C6烷硫基、C1-C6烷胺基、二(C1-C6烷基)胺基、C3-C8环烷基、C2-C8烯基、C2-C8炔基、芳基、含有1-3个杂原子的5-6元杂芳基或含有1-3个杂原子的3-10元杂环基，所述烷基、烷氧基、烷胺基、烷硫基、环烷基、芳基、杂芳基、杂环基任选被选自卤素、氰基、C1-C3烷基或C1-C3烷氧基的基团取代；当R3-R5、R7-R9为多个的情况下，任意相邻两个可结合形成环；R 3 -R 5 and R 7 -R 9 are each independently selected from: hydrogen, R 13 , halogen, cyano, amino, hydroxyl, mercapto, nitro, -R 21 N(R 22 )R 22, -R 21 C(=O)R 22 , -R 21 NHC(=O)R 22 , -R 21 C(=O)NHR 22 , -R 21 C(=O)OR 22 , -R 21 OC(=O)R 22 , -R 21 S(＝O) 2 R 22 , -R 21 S(＝O) 2 NHR 22 , C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkylthio group, C 1 -C 6 alkylamino group, di(C 1 -C 6 alkyl)amine group, C 3 -C 8 cycloalkyl group, C 2 -C 8 alkenyl group, C 2 -C 8 alkynyl group, aromatic group base, a 5-6-membered heteroaryl group containing 1-3 heteroatoms or a 3-10-membered heterocyclic group containing 1-3 heteroatoms, the alkyl group, alkoxy group, alkylamino group, alkylthio group , cycloalkyl, aryl, heteroaryl, heterocyclyl optionally substituted with a group selected from halogen, cyano, C 1 -C 3 alkyl or C 1 -C 3 alkoxy; when R 3 - When there are multiple R 5 , R 7 -R 9 , any two adjacent ones can be combined to form a ring;
R10选自：氢、卤素、氰基、氨基、硝基、-R21C(＝O)R22、-R21NHC(＝O)R22、-R21C(＝O)NHR22、-R21C(＝O)OR22、-R21OC(＝O)R22、-R21S(＝O)2R22、-R21S(＝O)2NHR22、C1-C6烷基、C1-C6烷氧基、C1-C6烷硫基、C1-C6烷胺基、二(C1-C6烷基)胺基、C3-C8环烷基、C2-C8烯基、C2-C8炔基、芳基、含有1-3个杂原子的5-6元杂芳基或含有1-3个杂原子的3-10元杂环基，所述烷基、烷氧基、烷胺基、烷硫基、环烷基、芳基、杂芳基、杂环基任选被选自卤素、氰基、C1-C3烷基或C1-C3烷氧基的基团取代；当R10为多个的情况下，任意相邻两个可结合形成环；R 10 is selected from: hydrogen, halogen, cyano, amino, nitro, -R 21 C(=O)R 22 , -R 21 NHC(=O)R 22 , -R 21 C(=O)NHR 22 , -R 21 C(=O)OR 22 , -R 21 OC(=O)R 22 , -R 21 S(=O) 2 R 22 , -R 21 S(=O) 2 NHR 22 , C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkylthio, C 1 -C 6 alkylamino, di(C 1 -C 6 alkyl)amine, C 3 -C 8 ring Alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, aryl, 5-6 membered heteroaryl containing 1-3 heteroatoms or 3-10 membered containing 1-3 heteroatoms Heterocyclyl group, the alkyl group, alkoxy group, alkylamino group, alkylthio group, cycloalkyl group, aryl group, heteroaryl group, heterocyclyl group are optionally selected from halogen, cyano group, C 1 -C 3 Alkyl or C 1 -C 3 alkoxy group substitution; when there are multiple R 10s , any two adjacent ones can be combined to form a ring;
R13选自：羟基、巯基、氨基、Rn、-ORn、-C(＝O)Rn、-C(＝O)ORn、-OC(＝O)Rn、-NRmRn、-C(＝O)NRmRn、-NRm-C(＝O)Rn、-NRn-C(＝O)Rm、-S(＝O)2Rn、-S(＝O)2NRmRn、-NRm-S(＝O)2Rn、-NRn-S(＝O)2Rm； R 13 is selected from: hydroxyl, mercapto, amino, R n , -OR n , -C(=O)R n , -C(=O)OR n , -OC(=O)R n , -NR m R n , -C(=O)NR m R n , -NR m -C(=O)R n , -NR n -C(=O)R m , -S(=O) 2 R n , -S(= O) 2 NR m R n , -NR m -S(=O) 2 R n , -NR n -S(=O) 2 R m ;
R14选自：Rn、-C(＝O)Rn、-C(＝O)ORn、-C(＝O)NRmRn、-S(＝O)2Rn、-S(＝O)2NRmRn；R 14 is selected from: R n , -C(=O)R n , -C(=O)OR n , -C(=O)NR m R n , -S(=O) 2 R n , -S( ＝O) 2 NR m R n ;
Rn选自：被羟基、巯基或氨基取代的C1-C8烷基，所述C1-C8烷基中的任意两个C-C之间可以插入选自-O-、-S-、-C(＝O)-、-C(＝O)O-、-OC(＝O)O-、-S(＝O)-、-S(＝O)2-、-NH-、-NHC(＝O)-、-NHC(＝O)NH-、-NHS(＝O)2-中的一种的基团，所述C1-C8烷基任选被1-3个分别选自卤素、氰基、氨基、硝基或C1-C3烷氧基的基团取代；R n is selected from: C 1 -C 8 alkyl group substituted by hydroxyl, mercapto or amino group. Any two CC in the C 1 -C 8 alkyl group can be inserted between -O-, -S-, -C(=O)-, -C(=O)O-, -OC(=O)O-, -S(=O)-, -S(=O) 2 -, -NH-, -NHC( =O)-, -NHC(=O)NH-, -NHS(=O) 2 -, the C 1 -C 8 alkyl group is optionally selected from 1 to 3 halogen groups. , cyano, amino, nitro or C 1 -C 3 alkoxy group substitution;
Rm独立地选自：氢、C1-C8烷基，所述C1-C8烷基中的任意两个C-C之间可以插入选自-O-、-S-、-C(＝O)-、-C(＝O)O-、-OC(＝O)O-、-S(＝O)-、-S(＝O)2-、-NH-、-NHC(＝O)-、-NHC(＝O)NH-、-NHS(＝O)2-中的一种的基团，所述C1-C8烷基任选被1-3个分别选自卤素、羟基、氰基、氨基、硝基或C1-C3烷氧基的基团取代；R m is independently selected from: hydrogen, C 1 -C 8 alkyl group, and any two CC in the C 1 -C 8 alkyl group can be inserted between any two CCs selected from -O-, -S-, -C(= O)-, -C(=O)O-, -OC(=O)O-, -S(=O)-, -S(=O) 2 -, -NH-, -NHC(=O)- , -NHC(=O)NH-, -NHS(=O) 2 -, the C 1 -C 8 alkyl group is optionally selected from 1-3 halogen, hydroxyl, cyanide, etc. Substituted with a base, amino, nitro or C 1 -C 3 alkoxy group;
R6选自：氢、C1-C6烷基、C3-C8环烷基、C2-C8烯基、C2-C8炔基、芳基、含有1-3个杂原子的5-6元杂芳基或含有1-3个杂原子的3-10元杂环基，所述烷基、环烷基、芳基、杂芳基、杂环基任选被1-3个分别选自卤素、羟基、氰基、氨基、硝基、-C(＝O)OR23、-OC(＝O)R23、-NHC(＝O)R23、-C(＝O)NHR23、C1-C3烷基、C1-C3烷氧基或-OP(＝O)(OM)2的基团取代；R 6 is selected from: hydrogen, C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, aryl, containing 1-3 heteroatoms 5-6 membered heteroaryl or 3-10 membered heterocyclic group containing 1-3 heteroatoms, the alkyl group, cycloalkyl group, aryl group, heteroaryl group and heterocyclic group are optionally replaced by 1-3 are selected from halogen, hydroxyl, cyano, amino, nitro, -C(=O)OR 23 , -OC(=O)R 23 , -NHC(=O)R 23 , -C(=O)NHR 23. Group substitution of C 1 -C 3 alkyl, C 1 - C 3 alkoxy or -OP(=O)(OM) 2 ;
M独立地选自：氢、C1-C4烷基；M is independently selected from: hydrogen, C 1 -C 4 alkyl;
R11、R12分别独立地选自：氢、C1-C4烷基、芳基、芳基-烷基；R 11 and R 12 are each independently selected from: hydrogen, C 1 -C 4 alkyl, aryl, aryl-alkyl;
R21选自：化学键、C1-C4亚烷基；R 21 is selected from: chemical bond, C 1 -C 4 alkylene;
R22、R23分别独立地选自：氢、C1-C4烷基、芳基、芳基-烷基，所述C1-C4烷基、芳基、芳基-烷基任选被选自卤素、羟基、氨基的基团取代；R 22 and R 23 are each independently selected from: hydrogen, C 1 -C 4 alkyl, aryl, aryl-alkyl, the C 1 -C 4 alkyl, aryl, aryl-alkyl is optional Substituted with a group selected from halogen, hydroxyl, and amino;
a选自：0、1、2、3、4或5；a is selected from: 0, 1, 2, 3, 4 or 5;
b选自：0、1、2、3、4或5；b is selected from: 0, 1, 2, 3, 4 or 5;
n选自：0、1、2或3；n is selected from: 0, 1, 2 or 3;
m选自：0、1、2、3或4；m is selected from: 0, 1, 2, 3 or 4;
o选自：1或2。oSelected from: 1 or 2.
优选地，所述式III所示的结构单元中至少包含一个选自R13或R14的基团。Preferably, the structural unit represented by formula III contains at least one group selected from R 13 or R 14 .
在一些实施方案中，D为式III所示的结构单元，D通过其含有的氧原子、或氮原子与L4相连；
In some embodiments, D is a structural unit represented by Formula III, and D is connected to L 4 through the oxygen atom or nitrogen atom it contains;
其中，in,
R1选自：Ra(CH2)a-、Ra(CH2)aC(＝O)-、Ra(CH2)aNHC(＝O)-、Ra(CH2)aOC(＝O)-或Ra(CH2)aS(＝O)2-，其中“CH2”任选被一个或多个R9取代，或者任选被-CH2CH2-取代，其中“NH”任选被R14取代；R 1 is selected from: R a (CH 2 ) a -, R a (CH 2 ) a C (=O)-, R a (CH 2 ) a NHC (= O) -, R a (CH 2 ) a OC (=O)- or R a (CH 2 ) a S(=O) 2 -, where "CH 2 " is optionally substituted by one or more R 9 , or optionally substituted by -CH 2 CH 2 -, where "NH" is optionally substituted by R 14 ;
Q选自：-NR2-、-O-、 Q is selected from: -NR 2 -, -O-,
R2选自：氢、Rb-、RbC(＝O)-、RbS(＝O)2-、R14；R 2 is selected from: hydrogen, R b -, R b C(=O)-, R b S(=O) 2 -, R 14 ;
环A表示至少含有一个N原子作为杂原子的杂环烷基，其通过N原子与T相连，环A任选被一个或多个选自R9的基团取代；Ring A represents a heterocycloalkyl group containing at least one N atom as a heteroatom, which is connected to T through an N atom. Ring A is optionally substituted by one or more groups selected from R 9 ;
Ra、Rb选自：C1-C8烷基、C3-C10环烷基、C3-C10桥环基、-NR11R12、C3-C10任含O、S、SO2、N或NHC(＝O)R22的杂环基、芳基、杂芳基、稠合芳基环烷基、稠合芳基杂环基、稠合杂芳基环烷基、稠合杂芳基杂环基、芳基-烷基、杂芳基-烷基、芳基-环烷基、芳基-杂环基、环烷基-杂环基、杂环基-杂环基，所述C1-C8烷基中的任意两个C-C之间可以插入选自-O-、-S-、-C(＝O)-、-S(＝O)-、-S(＝O)2-、-NH-、-NHC(＝O)-、-NHC(＝O)NH-、-NHS(＝O)2-中的一种的基团，Ra任选被一个或多个R9取代，Rb任选被一个或多个R10取代；R a and R b are selected from: C 1 -C 8 alkyl, C 3 -C 10 cycloalkyl, C 3 -C 10 bridged cyclic group, -NR 11 R 12 , C 3 -C 10 optionally containing O and S , SO 2 , N or NHC(=O)R 22 heterocyclyl, aryl, heteroaryl, fused arylcycloalkyl, fused arylheterocyclyl, fused heteroarylcycloalkyl, Fused heteroarylheterocyclyl, aryl-alkyl, heteroaryl-alkyl, aryl-cycloalkyl, aryl-heterocyclyl, cycloalkyl-heterocyclyl, heterocyclyl-heterocycle group, any two CCs in the C 1 -C 8 alkyl group can be inserted between any two CCs selected from -O-, -S-, -C(=O)-, -S(=O)-, -S( =O) 2 -, -NH-, -NHC(=O)-, -NHC(=O)NH-, -NHS(=O) 2 -, R a is optionally replaced by one or Multiple R 9 is substituted, and R b is optionally substituted by one or more R 10 ;
T、U、Z分别独立地选自：化学键、羰基、C1-C6亚烷基、C3-C10亚环烷基、亚芳基、亚杂芳基、亚杂环基，所述亚烷基、亚环烷基、亚芳基、亚杂芳基、亚杂环基任选被一个或多个R9取代；T, U, and Z are each independently selected from: chemical bond, carbonyl group, C 1 -C 6 alkylene group, C 3 -C 10 cycloalkylene group, arylene group, heteroarylene group, heterocyclylene group, the above Alkylene, cycloalkylene, arylene, heteroarylene, heterocyclylene are optionally substituted by one or more R 9 ;
Y选自：化学键、-C(＝O)NH(CH2)b-、-NHC(＝O)(CH2)b-、-O(CH2)b-、-NR2(CH2)b-，其中“CH2”任选被一个或多个R9取代，或者任选被-CH2CH2-取代，其中“NH”任选被R14取代；Y is selected from: chemical bond, -C(=O)NH(CH 2 ) b -, -NHC(=O)(CH 2 ) b -, -O(CH 2 ) b -, -NR 2 (CH 2 ) b -, wherein "CH 2 " is optionally substituted by one or more R 9 , or optionally substituted by -CH 2 CH 2 -, wherein "NH" is optionally substituted by R 14 ;
X选自：-(CR7R8)o-、-C(＝O)-；X is selected from: -(CR 7 R 8 ) o -, -C(=O)-;
R3-R5、R7-R9分别独立地选自：氢、R13、卤素、氰基、氨基、硝基、-R21C(＝O)R22、-R21NHC(＝O)R22、-R21C(＝O)NHR22、-R21C(＝O)OR22、-R21OC(＝O)R22、 -R21S(＝O)2R22、-R21S(＝O)2NHR22、C1-C6烷基、C1-C6烷氧基、C1-C6烷硫基、C1-C6烷胺基、二(C1-C6烷基)胺基、C3-C8环烷基、C2-C8烯基、C2-C8炔基、芳基、含有1-3个杂原子的5-6元杂芳基或含有1-3个杂原子的3-10元杂环基，所述烷基、烷氧基、烷胺基、烷硫基、环烷基、芳基、杂芳基、杂环基任选被选自卤素、氰基、C1-C3烷基或C1-C3烷氧基的基团取代；当R3-R5、R7-R9为多个的情况下，任意相邻两个可结合形成环；R 3 -R 5 and R 7 -R 9 are each independently selected from: hydrogen, R 13 , halogen, cyano, amino, nitro, -R 21 C(=O)R 22 , -R 21 NHC(=O )R 22 , -R 21 C(=O)NHR 22 , -R 21 C(=O)OR 22 , -R 21 OC(=O)R 22 , -R 21 S(=O) 2 R 22 , -R 21 S(=O) 2 NHR 22 , C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkylthio, C 1 -C 6 alkylamino, di(C 1 -C 6 alkyl)amine, C 3 -C 8 cycloalkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, aryl, A 5-6-membered heteroaryl group containing 1-3 heteroatoms or a 3-10-membered heterocyclic group containing 1-3 heteroatoms, the alkyl group, alkoxy group, alkylamino group, alkylthio group, ring Alkyl, aryl, heteroaryl, heterocyclyl are optionally substituted with a group selected from halogen, cyano, C 1 -C 3 alkyl or C 1 -C 3 alkoxy; when R 3 -R 5 , when R 7 to R 9 are multiple, any two adjacent ones can be combined to form a ring;
R10选自：氢、卤素、氰基、氨基、硝基、-R21C(＝O)R22、-R21NHC(＝O)R22、-R21C(＝O)NHR22、-R21C(＝O)OR22、-R21OC(＝O)R22、-R21S(＝O)2R22、-R21S(＝O)2NHR22、C1-C6烷基、C1-C6烷氧基、C1-C6烷硫基、C1-C6烷胺基、二(C1-C6烷基)胺基、C3-C8环烷基、C2-C8烯基、C2-C8炔基、芳基、含有1-3个杂原子的5-6元杂芳基或含有1-3个杂原子的3-10元杂环基，所述烷基、烷氧基、烷胺基、烷硫基、环烷基、芳基、杂芳基、杂环基任选被选自卤素、氰基、C1-C3烷基或C1-C3烷氧基的基团取代；当R10为多个的情况下，任意相邻两个可结合形成环；R 10 is selected from: hydrogen, halogen, cyano, amino, nitro, -R 21 C(=O)R 22 , -R 21 NHC(=O)R 22 , -R 21 C(=O)NHR 22 , -R 21 C(=O)OR 22 , -R 21 OC(=O)R 22 , -R 21 S(=O) 2 R 22 , -R 21 S(=O) 2 NHR 22 , C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkylthio, C 1 -C 6 alkylamino, di(C 1 -C 6 alkyl)amine, C 3 -C 8 ring Alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, aryl, 5-6 membered heteroaryl containing 1-3 heteroatoms or 3-10 membered containing 1-3 heteroatoms Heterocyclyl group, the alkyl group, alkoxy group, alkylamino group, alkylthio group, cycloalkyl group, aryl group, heteroaryl group, heterocyclyl group are optionally selected from halogen, cyano group, C 1 -C 3 Alkyl or C 1 -C 3 alkoxy group substitution; when there are multiple R 10s , any two adjacent ones can be combined to form a ring;
R13选自：羟基、Rn、-ORn、-C(＝O)Rn、-C(＝O)ORn、-OC(＝O)Rn、-NRmRn、-C(＝O)NRmRn、-NRm-C(＝O)Rn、-NRn-C(＝O)Rm、-S(＝O)2Rn、-S(＝O)2NRmRn、-NRm-S(＝O)2Rn、-NRn-S(＝O)2Rm；R 13 is selected from: hydroxyl, R n , -OR n , -C(=O)R n , -C(=O)OR n , -OC(=O)R n , -NR m R n , -C( =O)NR m R n , -NR m -C(=O)R n , -NR n -C(=O)R m , -S(=O) 2 R n , -S(=O) 2 NR m R n , -NR m -S(=O) 2 R n , -NR n -S(=O) 2 R m ;
R14选自：Rn、-C(＝O)Rn、-C(＝O)ORn、-C(＝O)NRmRn、-S(＝O)2Rn、-S(＝O)2NRmRn；R 14 is selected from: R n , -C(=O)R n , -C(=O)OR n , -C(=O)NR m R n , -S(=O) 2 R n , -S( ＝O) 2 NR m R n ;
Rn选自：被羟基取代的C1-C8烷基，所述C1-C8烷基中的任意两个C-C之间可以插入选自-O-、-S-、-C(＝O)-、-C(＝O)O-、-OC(＝O)O-、-S(＝O)-、-S(＝O)2-、-NH-、-NHC(＝O)-、-NHC(＝O)NH-、-NHS(＝O)2-中的一种的基团，所述C1-C8烷基任选被1-3个分别选自卤素、氰基、氨基、硝基或C1-C3烷氧基的基团取代；R n is selected from: C 1 -C 8 alkyl groups substituted by hydroxyl groups, and any two CCs in the C 1 -C 8 alkyl groups can be inserted between -O-, -S-, -C(= O)-, -C(=O)O-, -OC(=O)O-, -S(=O)-, -S(=O) 2 -, -NH-, -NHC(=O)- , -NHC(=O)NH-, -NHS(=O) 2 -, the C 1 -C 8 alkyl group is optionally selected from 1-3 halogen, cyano, Substitution of amino, nitro or C 1 -C 3 alkoxy groups;
Rm独立地选自：氢、C1-C8烷基，所述C1-C8烷基中的任意两个C-C之间可以插入选自-O-、-S-、-C(＝O)-、-C(＝O)O-、-OC(＝O)O-、-S(＝O)-、-S(＝O)2-、-NH-、-NHC(＝O)-、-NHC(＝O)NH-、-NHS(＝O)2-中的一种的基团，所述C1-C8烷基任选被1-3个分别选自卤素、羟基、氰基、氨基、硝基或C1-C3烷氧基的基团取代；R m is independently selected from: hydrogen, C 1 -C 8 alkyl group, and any two CC in the C 1 -C 8 alkyl group can be inserted between any two CCs selected from -O-, -S-, -C(= O)-, -C(=O)O-, -OC(=O)O-, -S(=O)-, -S(=O) 2 -, -NH-, -NHC(=O)- , -NHC(=O)NH-, -NHS(=O) 2 -, the C 1 -C 8 alkyl group is optionally selected from 1-3 halogen, hydroxyl, cyanide, etc. Substituted with a base, amino, nitro or C 1 -C 3 alkoxy group;
R6选自：氢、C1-C6烷基、C3-C8环烷基、C2-C8烯基、C2-C8炔基、芳基、含有1-3个杂原子的5-6元杂芳基或含有1-3个杂原子的3-10元杂环基，所述烷基、环烷基、芳基、杂芳基、杂环基任选被1-3个分别选自卤素、羟基、氰基、 氨基、硝基、-C(＝O)OR23、-OC(＝O)R23、-NHC(＝O)R23、-C(＝O)NHR23、C1-C3烷基、C1-C3烷氧基或-OP(＝O)(OM)2的基团取代；R 6 is selected from: hydrogen, C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, aryl, containing 1-3 heteroatoms 5-6 membered heteroaryl or 3-10 membered heterocyclic group containing 1-3 heteroatoms, the alkyl group, cycloalkyl group, aryl group, heteroaryl group and heterocyclic group are optionally replaced by 1-3 are selected from halogen, hydroxyl, cyano, Amino, nitro, -C(=O)OR 23 , -OC(=O)R 23 , -NHC(=O)R 23 , -C(=O)NHR 23 , C 1 -C 3 alkyl, C 1- C 3 alkoxy or -OP(=O)(OM) 2 group substitution;
M独立地选自：氢、C1-C4烷基；M is independently selected from: hydrogen, C 1 -C 4 alkyl;
R11、R12分别独立地选自：氢、C1-C4烷基、芳基、芳基-烷基；R 11 and R 12 are each independently selected from: hydrogen, C 1 -C 4 alkyl, aryl, aryl-alkyl;
R21选自：化学键、C1-C4亚烷基；R 21 is selected from: chemical bond, C 1 -C 4 alkylene;
R22、R23分别独立地选自：氢、C1-C4烷基、芳基、芳基-烷基，所述C1-C4烷基、芳基、芳基-烷基任选被选自卤素、羟基、氨基的基团取代；R 22 and R 23 are each independently selected from: hydrogen, C 1 -C 4 alkyl, aryl, aryl-alkyl, the C 1 -C 4 alkyl, aryl, aryl-alkyl is optional Substituted with a group selected from halogen, hydroxyl, and amino;
a选自：0、1、2、3、4或5；a is selected from: 0, 1, 2, 3, 4 or 5;
b选自：0、1、2、3、4或5；b is selected from: 0, 1, 2, 3, 4 or 5;
n选自：0、1、2或3；n is selected from: 0, 1, 2 or 3;
m选自：0、1、2、3或4；m is selected from: 0, 1, 2, 3 or 4;
o选自：1或2；oSelected from: 1 or 2;
其中，式III所示的结构单元中至少包含一个选自R13或R14的基团。Wherein, the structural unit represented by formula III contains at least one group selected from R 13 or R 14 .
优选地，所述Rn选自：羟基取代的C1-C8烷基；更优选的，所述Rn选自：羟基取代的C1-C4烷基；最优选地，所述Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH或-CH2CH2CH2CH2OH。优选地，Rm选自：氢、-C1-C4烷基。在一些实施方案中，所述Rn选自：羟基、巯基或氨基取代的C1-C8烷基；优选的，所述Rn选自：羟基、巯基或氨基取代的C1-C4烷基；更优选的，所述Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH、-CH2CH2CH2CH2OH、-CH2NH2、-CH2CH2NH2、-CH2CH2CH2NH2、-CH2CH2CH2CH2NH2、-CH2SH、-CH2CH2SH、-CH2CH2CH2SH、-CH2CH2CH2CH2SH。优选地，Rm选自：氢、-C1-C4烷基。Preferably, the R n is selected from: hydroxyl-substituted C 1 -C 8 alkyl; more preferably, the R n is selected from: hydroxyl substituted C 1 -C 4 alkyl; most preferably, the R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH or -CH 2 CH 2 CH 2 CH 2 OH. Preferably, R m is selected from: hydrogen, -C 1 -C 4 alkyl. In some embodiments, the R n is selected from: hydroxyl, mercapto or amino-substituted C 1 -C 8 alkyl; preferably, the R n is selected from: hydroxyl, mercapto or amino substituted C 1 -C 4 Alkyl group; more preferably, the R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 CH 2 SH. Preferably, R m is selected from: hydrogen, -C 1 -C 4 alkyl.
在一些实施方案中，D为式III所示的结构单元，D通过R13或R14的基团中的任何一个羟基与L4相连。在一些实施方案中，D为式III所示的结构单元，D通过R13或R14的基团中的任何一个羟基、氨基或巯基与L4相连。In some embodiments, D is a structural unit represented by Formula III, and D is connected to L 4 through any hydroxyl group in R 13 or R 14 . In some embodiments, D is a structural unit represented by Formula III, and D is connected to L 4 through any hydroxyl, amino or thiol group in R 13 or R 14 .
在一些实施方案中，所述R1选自：RaC(＝O)-、RaCH2C(＝O)-、RaCH2CH2C(＝O)-、Ra-、RaCH2-、RaCH2CH2-、RaNHC(＝O)-、RaCH2NHC(＝O)-、RaCH2CH2NHC(＝O)-、RaOC(＝O)-、RaCH2OC(＝O)-、RaCH2CH2OC(＝O)-、RaS(＝O)2-、RaCH2S(＝O)2-、RaCH2CH2S(＝O)2-，其中“CH2”任选被一个或多个R9取代，或者任选被-CH2CH2-取代，其中“NH”任选被R14取代。 In some embodiments, the R 1 is selected from: Ra C(=O)-, Ra CH 2 C(=O)-, Ra CH 2 CH 2 C (=O)-, Ra - , R a CH 2 -, R a CH 2 CH 2 -, R a NHC(=O)-, R a CH 2 NHC(=O)-, R a CH 2 CH 2 NHC(=O)-, R a OC (=O)-, R a CH 2 OC (= O) -, R a CH 2 CH 2 OC (= O) -, R a S (= O) 2 -, R a CH 2 S (= O) 2 -, R a CH 2 CH 2 S(=O) 2 -, where "CH 2 " is optionally substituted by one or more R 9 , or optionally substituted by -CH 2 CH 2 -, where "NH" is optional Replaced by R 14 .
优选地，所述R1选自：RaC(＝O)-、RaCHR9C(＝O)-、RaCHR9CH2C(＝O)-、RaCHCHR9C(＝O)-、Ra-、RaCHR9-、RaCHR9CH2-、RaCHCHR9-、RaNR14C(＝O)-、RaCHR9NHC(＝O)-、RaCHNR14C(＝O)-、RaCHR9CH2NHC(＝O)-、RaCH2CHR9NHC(＝O)-、RaCH2CH2NR14C(＝O)-、RaOC(＝O)-、RaCHR9OC(＝O)-、RaCHR9CH2OC(＝O)-、RaCH2CHR9OC(＝O)-、RaCHR9S(＝O)2-、RaCHR9CH2S(＝O)2-、RaCH2CHR9S(＝O)2-。Preferably, the R 1 is selected from: Ra C(=O)-, Ra CHR 9 C(=O)-, Ra CHR 9 CH 2 C(=O)-, Ra CHCHR 9 C(= O)-, R a -, R a CHR 9 -, R a CHR 9 CH 2 -, R a CHCHR 9 -, R a NR 14 C(=O)-, R a CHR 9 NHC(=O)-, R a CHNR 14 C (= O) -, R a CHR 9 CH 2 NHC ( = O) -, R a CH 2 CHR 9 NHC ( = O) -, R a CH 2 CH 2 NR 14 C ( = O) -, R a OC(=O)-, R a CHR 9 OC(=O)-, R a CHR 9 CH 2 OC(=O)-, R a CH 2 CHR 9 OC(=O)-, R a CHR 9 S(=O) 2 -, Ra CHR 9 CH 2 S(=O) 2 -, Ra CH 2 CHR 9 S(=O) 2 -.
优选地，所述R9选自：氢、C1-C4烷基、R13；更优选地，所述R9选自：R13。在一些实施方案中，所述R9选自：氢、羟基、巯基、氨基、C1-C4烷基、-R21N(R22)R22、-R21C(＝O)R22、R13；更优选地，所述R9选自：羟基、巯基、氨基、-R21N(R22)R22、-R21C(＝O)R22、R13。Preferably, the R 9 is selected from: hydrogen, C 1 -C 4 alkyl, R 13 ; more preferably, the R 9 is selected from: R 13 . In some embodiments, the R 9 is selected from: hydrogen, hydroxyl, thiol, amino, C 1 -C 4 alkyl, -R 21 N(R 22 )R 22 , -R 21 C(=O)R 22 , R 13 ; more preferably, the R 9 is selected from: hydroxyl, mercapto, amino, -R 21 N(R 22 )R 22 , -R 21 C(=O)R 22 , R 13 .
优选地，所述R13选自：羟基、Rn、-ORn。在一些实施方案中，所述R13选自：羟基、巯基、氨基、Rn、-ORn。Preferably, the R 13 is selected from: hydroxyl, R n , -OR n . In some embodiments, the R 13 is selected from: hydroxyl, thiol, amino, R n , -OR n .
优选地，所述R14选自：Rn、-C(＝O)Rn。Preferably, the R 14 is selected from: R n , -C(=O)R n .
优选地，所述Rn选自：羟基取代的C1-C8烷基；更优选地，所述Rn选自：羟基取代的C1-C4烷基；最优选地，所述Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH或-CH2CH2CH2CH2OH。在一些实施方案中，所述Rn选自：羟基、巯基或氨基取代的C1-C8烷基；优选的，所述Rn选自：羟基、巯基或氨基取代的C1-C4烷基；更优选的，所述Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH、-CH2CH2CH2CH2OH、-CH2NH2、-CH2CH2NH2、-CH2CH2CH2NH2、-CH2CH2CH2CH2NH2、-CH2SH、-CH2CH2SH、-CH2CH2CH2SH、-CH2CH2CH2CH2SH。Preferably, the R n is selected from: hydroxyl-substituted C 1 -C 8 alkyl; more preferably, the R n is selected from: hydroxyl-substituted C 1 -C 4 alkyl; most preferably, the R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH or -CH 2 CH 2 CH 2 CH 2 OH. In some embodiments, the R n is selected from: hydroxyl, mercapto or amino-substituted C 1 -C 8 alkyl; preferably, the R n is selected from: hydroxyl, mercapto or amino substituted C 1 -C 4 Alkyl group; more preferably, the R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 CH 2 SH.
在一些实施方案中，所述Ra选自：苯基、萘基、咔唑基、1-氮杂咔唑基、2-氮杂咔唑基、1,8-二氮杂咔唑基、吲哚基、7-氮杂吲哚基、2,3-二氢吲哚基、2,3-二氢-7-氮杂吲哚基、吩噁嗪基、芴基、喹啉基、异喹啉基、萘啶基、四氢萘啶基、四氢喹啉基、嘧啶基、三唑基、双环[1.1.1]戊烷基、降冰片烷基、金刚烷基。优选地，所述Ra任选被一个或多个R9取代，所述R9选自氢、R13、卤素、氰基、C1-C6烷基、C1-C6卤代烷基、C1-C6烷氧基、苯基、萘基。在一些实施方案中，所述Ra选自：苯基、萘基、咔唑基、1-氮杂咔唑基、2-氮杂咔唑基、1,8-二氮杂咔唑基、吲哚基、7-氮杂吲哚基、2,3-二氢吲哚基、2,3-二氢-7-氮杂吲哚基、吩噁嗪基、芴基、喹啉基、异喹啉基、萘啶基、四氢萘啶基、四氢喹啉基、嘧啶基、吡啶基、喹啉基-吡啶基、三唑基、双环[1.1.1]戊烷基、降冰片烷基、金刚烷基。 优选的，所述Ra任选被一个或多个R9取代，所述R9选自氢、羟基、巯基、-R21N(R22)R22、-R21C(＝O)R22、R13、卤素、氰基、C1-C6烷基、C1-C6卤代烷基、C1-C6烷氧基、苯基、萘基。In some embodiments, the R a is selected from: phenyl, naphthyl, carbazolyl, 1-azacarbazolyl, 2-azacarbazolyl, 1,8-diazacarbazolyl, Indolyl, 7-azaindolyl, 2,3-dihydroindolyl, 2,3-dihydro-7-azaindolyl, phenoxazinyl, fluorenyl, quinolinyl, iso Quinolyl, naphthyridinyl, tetralinyl, tetrahydroquinolinyl, pyrimidinyl, triazolyl, bicyclo[1.1.1]pentyl, norbornyl, adamantyl. Preferably, the R a is optionally substituted by one or more R 9 , and the R 9 is selected from hydrogen, R 13 , halogen, cyano, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, C 1 -C 6 alkoxy, phenyl, naphthyl. In some embodiments, the R a is selected from: phenyl, naphthyl, carbazolyl, 1-azacarbazolyl, 2-azacarbazolyl, 1,8-diazacarbazolyl, Indolyl, 7-azaindolyl, 2,3-dihydroindolyl, 2,3-dihydro-7-azaindolyl, phenoxazinyl, fluorenyl, quinolinyl, iso Quinolyl, naphthyridinyl, tetralinyl, tetrahydroquinolinyl, pyrimidinyl, pyridyl, quinolyl-pyridyl, triazolyl, bicyclo[1.1.1]pentyl, norbornane base, adamantyl. Preferably, the R a is optionally substituted by one or more R 9 , and the R 9 is selected from hydrogen, hydroxyl, mercapto, -R 21 N(R 22 )R 22 , -R 21 C(=O)R 22. R 13 , halogen, cyano, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, C 1 -C 6 alkoxy, phenyl, naphthyl.
优选地，所述Ra选自：苯基、萘-1-基、萘-2-基、咔唑-9-基、1-氮杂咔唑-9-基、2-氮杂咔唑-9-基、1,8-二氮杂咔唑-9-基、吲哚-1-基、2,3-二氢-吲哚-1-基、7-氮杂吲哚基-1-基、2,3-二氢-7-氮杂吲哚基-1-基、吩噁嗪-10-基、芴-9-基、喹啉-4-基、喹啉-5-基、喹啉-8-基、异喹啉-1-基、异喹啉-4-基、异喹啉-5-基、异喹啉-8-基、1,2,3,4-四氢-1,8-萘啶-1-基、1,2,3,4-四氢喹啉-1-基、嘧啶-2-基、嘧啶-4-基、嘧啶-5-基、1,2,3-三唑-1-基、1,2,4-三唑-1-基、1,3,4-三唑-1-基、双环[1.1.1]戊烷-1-基、降冰片烷-1-基、金刚烷-1-基。优选地，所述Ra任选被一个或多个R9取代，所述R9选自：R13、F、Cl、Br、氰基、甲基、乙基、三氟甲基、甲氧基、乙氧基、苯基、萘-1-基、萘-2-基。在一些实施方案中，所述Ra选自：苯基、萘-1-基、萘-2-基、咔唑-9-基、1-氮杂咔唑-9-基、2-氮杂咔唑-9-基、1,8-二氮杂咔唑-9-基、吲哚-1-基、2,3-二氢-吲哚-1-基、7-氮杂吲哚基-1-基、2,3-二氢-7-氮杂吲哚基-1-基、吩噁嗪-10-基、芴-9-基、喹啉-4-基、喹啉-5-基、喹啉-8-基、异喹啉-1-基、异喹啉-4-基、异喹啉-5-基、异喹啉-8-基、1,2,3,4-四氢-1,8-萘啶-1-基、1,2,3,4-四氢喹啉-1-基、嘧啶-2-基、嘧啶-4-基、嘧啶-5-基、吡啶-2-基、吡啶-3-基、吡啶-4-基、5-(8-喹啉-基)-吡啶-2-基、1,2,3-三唑-1-基、1,2,4-三唑-1-基、1,3,4-三唑-1-基、双环[1.1.1]戊烷-1-基、降冰片烷-1-基、金刚烷-1-基。优选的，Ra任选被一个或多个R9取代，所述R9选自：R13、F、Cl、Br、氰基、甲基、乙基、三氟甲基、甲氧基、乙氧基、苯基、萘-1-基、萘-2-基。Preferably, the R a is selected from: phenyl, naphthyl-1-yl, naphthyl-2-yl, carbazol-9-yl, 1-azacarbazol-9-yl, 2-azacarbazole- 9-yl, 1,8-diazacarbazol-9-yl, indol-1-yl, 2,3-dihydro-indol-1-yl, 7-azaindol-1-yl , 2,3-dihydro-7-azaindolyl-1-yl, phenoxazin-10-yl, fluoren-9-yl, quinolin-4-yl, quinolin-5-yl, quinoline -8-yl, isoquinolin-1-yl, isoquinolin-4-yl, isoquinolin-5-yl, isoquinolin-8-yl, 1,2,3,4-tetrahydro-1, 8-naphthyridin-1-yl, 1,2,3,4-tetrahydroquinolin-1-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, 1,2,3-yl Triazol-1-yl, 1,2,4-triazol-1-yl, 1,3,4-triazol-1-yl, bicyclo[1.1.1]pentan-1-yl, norbornan- 1-yl, adamantane-1-yl. Preferably, the R a is optionally substituted by one or more R 9 , and the R 9 is selected from: R 13 , F, Cl, Br, cyano, methyl, ethyl, trifluoromethyl, methoxy base, ethoxy, phenyl, naphthyl-1-yl, naphthyl-2-yl. In some embodiments, the R a is selected from: phenyl, naphth-1-yl, naphth-2-yl, carbazol-9-yl, 1-azacarbazol-9-yl, 2-aza Carbazol-9-yl, 1,8-diazacarbazol-9-yl, indol-1-yl, 2,3-dihydro-indol-1-yl, 7-azaindolyl- 1-yl, 2,3-dihydro-7-azaindolyl-1-yl, phenoxazin-10-yl, fluoren-9-yl, quinolin-4-yl, quinolin-5-yl , quinolin-8-yl, isoquinolin-1-yl, isoquinolin-4-yl, isoquinolin-5-yl, isoquinolin-8-yl, 1,2,3,4-tetrahydro -1,8-naphthyridin-1-yl, 1,2,3,4-tetrahydroquinolin-1-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, pyridin-2 -yl, pyridin-3-yl, pyridin-4-yl, 5-(8-quinolin-yl)-pyridin-2-yl, 1,2,3-triazol-1-yl, 1,2,4 -Triazol-1-yl, 1,3,4-triazol-1-yl, bicyclo[1.1.1]pentan-1-yl, norbornan-1-yl, adamantan-1-yl. Preferably, R a is optionally substituted by one or more R 9 , and the R 9 is selected from: R 13 , F, Cl, Br, cyano, methyl, ethyl, trifluoromethyl, methoxy, Ethoxy, phenyl, naphthyl-1-yl, naphthyl-2-yl.
优选地，所述R13选自：羟基、Rn、-ORn、-OC(＝O)Rn、-NRmRn、-NRm-C(＝O)Rn、-NRn-C(＝O)Rm、-NRm-S(＝O)2Rn、-NRn-S(＝O)2Rm；更优选地，所述R13选自：羟基、Rn、-ORn、-NRmRn、-NRn-C(＝O)Rm、-NRn-S(＝O)2Rm。在一些实施方案中，所述R13选自：羟基、巯基、氨基、Rn、-ORn、-OC(＝O)Rn、-NRmRn、-NRm-C(＝O)Rn、-NRn-C(＝O)Rm、-NRm-S(＝O)2Rn、-NRn-S(＝O)2Rm；更优选的，所述R13选自：羟基、巯基、氨基、Rn、-ORn、-NRmRn、-NRn-C(＝O)Rm、-NRn-S(＝O)2Rm。Preferably, the R 13 is selected from: hydroxyl, R n , -OR n , -OC(=O)R n , -NR m R n , -NR m -C(=O)R n , -NR n - C(=O)R m , -NR m -S(=O) 2 R n , -NR n -S(=O) 2 R m ; more preferably, the R 13 is selected from: hydroxyl, R n , -OR n , -NR m R n , -NR n -C(=O)R m , -NR n -S(=O) 2 R m . In some embodiments, the R 13 is selected from: hydroxyl, thiol, amino, R n , -OR n , -OC(=O)R n , -NR m R n , -NR m -C(=O) R n , -NR n -C(=O)R m , -NR m -S(=O) 2 R n , -NR n -S(=O) 2 R m ; more preferably, the R 13 is selected from From: hydroxyl group, thiol group, amino group, R n , -OR n , -NR m R n , -NR n -C(=O)R m , -NR n -S(=O) 2 R m .
优选地，所述Rn选自：羟基取代的C1-C8烷基；更优选地，所述Rn选自：羟基取代的C1-C4烷基；最优选地，Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH或-CH2CH2CH2CH2OH。在一些实施方案中，所述Rn选自：羟 基、巯基或氨基取代的C1-C8烷基；优选的，所述Rn选自：羟基、巯基或氨基取代的C1-C4烷基；更优选的，所述Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH、-CH2CH2CH2CH2OH、-CH2NH2、-CH2CH2NH2、-CH2CH2CH2NH2、-CH2CH2CH2CH2NH2、-CH2SH、-CH2CH2SH、-CH2CH2CH2SH、-CH2CH2CH2CH2SH。Preferably, the R n is selected from: hydroxyl-substituted C 1 -C 8 alkyl; more preferably, the R n is selected from: hydroxyl substituted C 1 -C 4 alkyl; most preferably, R n is selected from From: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, or -CH 2 CH 2 CH 2 CH 2 OH. In some embodiments, the R n is selected from: hydroxyl hydroxyl, mercapto or amino-substituted C 1 -C 8 alkyl; preferably, the R n is selected from: hydroxyl, mercapto or amino substituted C 1 -C 4 alkyl; more preferably, the R n is selected from : -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 CH 2 SH .
优选地，所述Rm选自：氢、-C1-C4烷基。Preferably, the R m is selected from: hydrogen, -C 1 -C 4 alkyl.
优选地，所述R9选自：F、Cl、Br、氰基、甲基、乙基、三氟甲基、甲氧基、乙氧基、羟基、-CH2OH、-CH2CH2OH、-CH2CH2CH2OH、-OCH2OH、-OCH2CH2OH、-OCH2CH2CH2OH、-NHCH2OH、-NHCH2CH2OH、-NHCH2CH2CH2OH、-N(CH3)CH2OH、-N(CH3)CH2CH2OH、-N(CH3)CH2CH2CH2OH、-N(COCH3)CH2OH、-N(COCH3)CH2CH2OH、-N(COCH3)CH2CH2CH2OH、-N(SO2CH3)CH2OH、-N(SO2CH3)CH2CH2OH、-N(SO2CH3)CH2CH2CH2OH。在一些实施方案中，所述R9选自：F、Cl、Br、氰基、甲基、乙基、三氟甲基、甲氧基、乙氧基、羟基、巯基、氨基、-C(＝O)CH3、C(＝O)CH2CH3、C(＝O)CH2CH2CH3、-CH2CH2N(CH3)2、-CH2OH、-CH2CH2OH、-CH2CH2CH2OH、-CH2NH2、-CH2CH2NH2、-CH2CH2CH2NH2、-CH2SH、-CH2CH2SH、-CH2CH2CH2SH、-CH2CH2CH2CH2SH、-OCH2OH、-OCH2CH2OH、-OCH2CH2CH2OH、-OCH2NH2、-OCH2CH2NH2、-OCH2CH2CH2NH2、-OCH2SH、-OCH2CH2SH、-OCH2CH2CH2SH、-NHCH2OH、-NHCH2CH2OH、-NHCH2CH2CH2OH、-NHCH2NH2、-NHCH2CH2NH2、-NHCH2CH2CH2NH2、-NHCH2SH、-NHCH2CH2SH、-NHCH2CH2CH2SH、-N(CH3)CH2OH、-N(CH3)CH2CH2OH、-N(CH3)CH2CH2CH2OH、-N(CH3)CH2NH2、-N(CH3)CH2CH2NH2、-N(CH3)CH2CH2CH2NH2、-N(CH3)CH2SH、-N(CH3)CH2CH2SH、-N(CH3)CH2CH2CH2SH、-N(COCH3)CH2OH、-N(COCH3)CH2CH2OH、-N(COCH3)CH2CH2CH2OH、-N(COCH3)CH2NH2、-N(COCH3)CH2CH2NH2、-N(COCH3)CH2CH2CH2NH2、-N(COCH3)CH2SH、-N(COCH3)CH2CH2SH、-N(COCH3)CH2CH2CH2SH-N(SO2CH3)CH2OH、-N(SO2CH3)CH2CH2OH、-N(SO2CH3)CH2CH2CH2OH、-N(SO2CH3)CH2NH2、-N(SO2CH3)CH2CH2NH2、-N(SO2CH3)CH2CH2CH2NH2、-N(SO2CH3)CH2SH、-N(SO2CH3)CH2CH2SH、-N(SO2CH3)CH2CH2CH2SH。Preferably, the R 9 is selected from: F, Cl, Br, cyano, methyl, ethyl, trifluoromethyl, methoxy, ethoxy, hydroxyl, -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -OCH 2 OH, -OCH 2 CH 2 OH, -OCH 2 CH 2 CH 2 OH, -NHCH 2 OH, -NHCH 2 CH 2 OH, -NHCH 2 CH 2 CH 2 OH, -N(CH 3 )CH 2 OH, -N(CH 3 )CH 2 CH 2 OH, -N(CH 3 )CH 2 CH 2 CH 2 OH, -N(COCH 3 )CH 2 OH, - N(COCH 3 )CH 2 CH 2 OH, -N(COCH 3 )CH 2 CH 2 CH 2 OH, -N(SO 2 CH 3 )CH 2 OH, -N(SO 2 CH 3 )CH 2 CH 2 OH , -N(SO 2 CH 3 )CH 2 CH 2 CH 2 OH. In some embodiments, the R 9 is selected from: F, Cl, Br, cyano, methyl, ethyl, trifluoromethyl, methoxy, ethoxy, hydroxyl, thiol, amino, -C( ＝O)CH 3 , C(＝O)CH 2 CH 3 , C(＝O)CH 2 CH 2 CH 3 , -CH 2 CH 2 N(CH 3 ) 2 , -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 CH 2 SH, -OCH 2 OH, -OCH 2 CH 2 OH, -OCH 2 CH 2 CH 2 OH, -OCH 2 NH 2 , -OCH 2 CH 2 NH 2. -OCH 2 CH 2 CH 2 NH 2 , -OCH 2 SH , -OCH 2 CH 2 SH , -OCH 2 CH 2 CH 2 SH , -NHCH 2 OH , -NHCH 2 CH 2 OH , -NHCH 2 CH 2 CH 2 OH, -NHCH 2 NH 2 , -NHCH 2 CH 2 NH 2 , -NHCH 2 CH 2 CH 2 NH 2 , -NHCH 2 SH, -NHCH 2 CH 2 SH, -NHCH 2 CH 2 CH 2 SH, - N(CH 3 )CH 2 OH, -N(CH 3 )CH 2 CH 2 OH, -N(CH 3 )CH 2 CH 2 CH 2 OH, -N(CH 3 )CH 2 NH 2 , -N(CH 3 )CH 2 CH 2 NH 2 , -N(CH 3 )CH 2 CH 2 CH 2 NH 2 , -N(CH 3 )CH 2 SH , -N(CH 3 )CH 2 CH 2 SH , -N(CH 3 )CH 2 CH 2 CH 2 SH , -N(COCH 3 )CH 2 OH, -N(COCH 3 )CH 2 CH 2 OH, -N(COCH 3 )CH 2 CH 2 CH 2 OH, -N(COCH 3 )CH 2 NH 2 , -N(COCH 3 )CH 2 CH 2 NH 2 , -N(COCH 3 )CH 2 CH 2 CH 2 NH 2 , -N(COCH 3 )CH 2 SH, -N(COCH 3 )CH 2 CH 2 SH, -N(COCH 3 )CH 2 CH 2 CH 2 SH -N(SO 2 CH 3 )CH 2 OH, -N(SO 2 CH 3 )CH 2 CH 2 OH, -N(SO 2 CH 3 )CH 2 CH 2 CH 2 OH, -N(SO 2 CH 3 )CH 2 NH 2 , -N(SO 2 CH 3 )CH 2 CH 2 NH 2 , -N(SO 2 CH 3 )CH 2 CH 2 CH 2 NH 2, -N(SO 2 CH 3 )CH 2 SH, -N(SO 2 CH 3 )CH 2 CH 2 SH, -N(SO 2 CH 3 )CH 2 CH 2 CH 2 SH.
优选地，所述Ra选自以下基团：
Preferably, said R a is selected from the following groups:
在一些实施方案中，所述Ra选自以下基团：
In some embodiments, the Ra is selected from the following groups:
在一些实施方案中，所述Q选自：-NR2-、 In some embodiments, the Q is selected from: -NR2- ,
优选地，所述R2选自：氢、R14、C1-C8烷基、C3-C10环烷基、芳基、C(＝O)C3-C10环烷基、-C(＝O)C1-C8烷基或-S(＝O)2C1-C8烷基，所述C1-C8烷基、C3-C10环烷基、 芳基以及C(＝O)C3-C10环烷基、-C(＝O)C1-C8烷基或-S(＝O)2C1-C8烷基的环烷基或烷基部分任选被一个或多个R9取代，所述R9选自：氢、卤素、氨基、氰基、羧基、C1-C6烷基、C1-C6烷氧基或芳基；更优选地，所述R2选自：R14。Preferably, the R 2 is selected from: hydrogen, R 14 , C 1 -C 8 alkyl, C 3 -C 10 cycloalkyl, aryl, C(=O)C 3 -C 10 cycloalkyl, - C(=O)C 1 -C 8 alkyl or -S(=O) 2 C 1 -C 8 alkyl, the C 1 -C 8 alkyl, C 3 -C 10 cycloalkyl, Aryl and C(=O)C 3 -C 10 cycloalkyl, -C(=O)C 1 -C 8 alkyl or -S(=O) 2 C 1 -C 8 alkyl cycloalkyl or The alkyl moiety is optionally substituted with one or more R 9 selected from: hydrogen, halogen, amino, cyano, carboxyl , C 1 -C 6 alkyl, C 1 -C 6 alkoxy or aromatic group; more preferably, the R 2 is selected from: R 14 .
优选地，所述R14选自：Rn、-C(＝O)Rn、-C(＝O)ORn、-C(＝O)NRmRn、-S(＝O)2Rn、-S(＝O)2NRmRn。Preferably, the R 14 is selected from: R n , -C(=O)R n , -C(=O)OR n , -C(=O)NR m R n , -S(=O) 2 R n , -S(=O) 2 NR m R n .
优选地，所述Rn选自：羟基取代的C1-C8烷基；更优选地，所述Rn选自：羟基取代的C1-C4烷基；最优选地，所述Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH或-CH2CH2CH2CH2OH。在一些实施方案中，所述Rn选自：羟基、巯基或氨基取代的C1-C8烷基；更优选的，所述Rn选自：羟基、巯基或氨基取代的C1-C4烷基；最优选的，Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH、-CH2CH2CH2CH2OH、-CH2NH2、-CH2CH2NH2、-CH2CH2CH2NH2、-CH2CH2CH2CH2NH2、-CH2SH、-CH2CH2SH、-CH2CH2CH2SH、-CH2CH2CH2CH2SH。Preferably, the R n is selected from: hydroxyl-substituted C 1 -C 8 alkyl; more preferably, the R n is selected from: hydroxyl-substituted C 1 -C 4 alkyl; most preferably, the R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH or -CH 2 CH 2 CH 2 CH 2 OH. In some embodiments, the R n is selected from: hydroxyl, mercapto or amino-substituted C 1 -C 8 alkyl; more preferably, the R n is selected from: hydroxyl, mercapto or amino substituted C 1 -C 4 alkyl; most preferably, R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 CH 2 SH.
优选地，所述Rm选自：氢、-C1-C4烷基。Preferably, the R m is selected from: hydrogen, -C 1 -C 4 alkyl.
优选地，所述R2选自：氢、甲基、乙基、-SO2CH3、-COCH3、-CO-异丙基、-CO-环丙基、异丙基、环丙基、2-甲氧基乙基、2-氰基乙基、苯基、萘基、苄基、2-苯基乙基、1-萘甲基、2-萘乙基、-CH2OH、-CH2CH2OH、-CH2CH2CH2OH、-C(＝O)CH2OH、-C(＝O)CH2CH2OH、-C(＝O)CH2CH2CH2OH、-C(＝O)OCH2OH、-C(＝O)OCH2CH2OH、-C(＝O)OCH2CH2CH2OH、-C(＝O)NHCH2OH、-C(＝O)NHCH2CH2OH、-C(＝O)NHCH2CH2CH2OH、-S(＝O)2CH2OH、-S(＝O)2CH2CH2OH、-S(＝O)2CH2CH2CH2OH、-S(＝O)2NHCH2OH、-S(＝O)2NHCH2CH2OH、-S(＝O)2NHCH2CH2CH2OH。在一些实施方案中，所述R2进一步选自：氢、甲基、乙基、-SO2CH3、-COCH3、-CO-异丙基、-CO-环丙基、异丙基、环丙基、2-甲氧基乙基、2-氰基乙基、苯基、萘基、苄基、2-苯基乙基、1-萘甲基、2-萘乙基、-CH2OH、-CH2CH2OH、-CH2CH2CH2OH、-CH2NH2、-CH2CH2NH2、-CH2CH2CH2NH2、-CH2SH、-CH2CH2SH、-CH2CH2CH2SH、-CH2CH2CH2CH2SH、-C(＝O)CH2OH、-C(＝O)CH2CH2OH、-C(＝O)CH2CH2CH2OH、-C(＝O)CH2NH2、-C(＝O)CH2CH2NH2、-C(＝O)CH2CH2CH2NH2、-C(＝O)CH2SH、-C(＝O)CH2CH2SH、-C(＝O)CH2CH2CH2SH、-C(＝O)OCH2OH、-C(＝O)OCH2CH2OH、-C(＝O)OCH2CH2CH2OH、-C(＝O)OCH2NH2、-C(＝O)OCH2CH2NH2、 -C(＝O)OCH2CH2CH2NH2、-C(＝O)OCH2SH、-C(＝O)OCH2CH2SH、-C(＝O)OCH2CH2CH2SH、-C(＝O)NHCH2OH、-C(＝O)NHCH2CH2OH、-C(＝O)NHCH2CH2CH2OH、-C(＝O)NHCH2NH2、-C(＝O)NHCH2CH2NH2、-C(＝O)NHCH2CH2CH2NH2、-C(＝O)NHCH2SH、-C(＝O)NHCH2CH2SH、-C(＝O)NHCH2CH2CH2SH、-S(＝O)2CH2OH、-S(＝O)2CH2CH2OH、-S(＝O)2CH2CH2CH2OH、-S(＝O)2CH2NH2、-S(＝O)2CH2CH2NH2、-S(＝O)2CH2CH2CH2NH2、-S(＝O)2CH2SH、-S(＝O)2CH2CH2SH、-S(＝O)2CH2CH2CH2SH、-S(＝O)2NHCH2OH、-S(＝O)2NHCH2CH2OH、-S(＝O)2NHCH2CH2CH2OH、-S(＝O)2NHCH2NH2、-S(＝O)2NHCH2CH2NH2、-S(＝O)2NHCH2CH2CH2NH2、-S(＝O)2NHCH2SH、-S(＝O)2NHCH2CH2SH、-S(＝O)2NHCH2CH2CH2SH。Preferably, the R 2 is selected from: hydrogen, methyl, ethyl, -SO 2 CH 3 , -COCH 3 , -CO-isopropyl, -CO-cyclopropyl, isopropyl, cyclopropyl, 2-methoxyethyl, 2-cyanoethyl, phenyl, naphthyl, benzyl, 2-phenylethyl, 1-naphthylmethyl, 2-naphthylethyl, -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -C(＝O)CH 2 OH, -C(＝O)CH 2 CH 2 OH, -C(＝O)CH 2 CH 2 CH 2 OH, -C(＝O)OCH 2 OH, -C(＝O)OCH 2 CH 2 OH, -C(＝O)OCH 2 CH 2 CH 2 OH, -C(＝O)NHCH 2 OH, -C(＝ O)NHCH 2 CH 2 OH, -C(＝O)NHCH 2 CH 2 CH 2 OH, -S(＝O) 2 CH 2 OH, -S(＝O) 2 CH 2 CH 2 OH, -S(＝ O) 2 CH 2 CH 2 CH 2 OH, -S(=O) 2 NHCH 2 OH, -S(=O) 2 NHCH 2 CH 2 OH, -S(=O) 2 NHCH 2 CH 2 CH 2 OH. In some embodiments, the R 2 is further selected from: hydrogen, methyl, ethyl, -SO 2 CH 3 , -COCH 3 , -CO-isopropyl, -CO-cyclopropyl, isopropyl, Cyclopropyl, 2-methoxyethyl, 2-cyanoethyl, phenyl, naphthyl, benzyl, 2-phenylethyl, 1-naphthylmethyl, 2-naphthylethyl, -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 CH 2 SH, -C(＝O)CH 2 OH, -C(＝O)CH 2 CH 2 OH, -C(＝ O)CH 2 CH 2 CH 2 OH, -C(＝O)CH 2 NH 2 , -C(＝O)CH 2 CH 2 NH 2 , -C(＝O)CH 2 CH 2 CH 2 NH 2 , - C(=O)CH 2 SH, -C(=O)CH 2 CH 2 SH, -C(=O)CH 2 CH 2 CH 2 SH, -C(=O)OCH 2 OH, -C(=O )OCH 2 CH 2 OH, -C(＝O)OCH 2 CH 2 CH 2 OH, -C(＝O)OCH 2 NH 2 , -C(＝O)OCH 2 CH 2 NH 2 , -C(＝O)OCH 2 CH 2 CH 2 NH 2 , -C(＝O)OCH 2 SH , -C(＝O)OCH 2 CH 2 SH , -C(＝O)OCH 2 CH 2 CH 2 SH , -C(＝O)NHCH 2 OH, -C(＝O)NHCH 2 CH 2 OH, -C(＝O)NHCH 2 CH 2 CH 2 OH, -C(＝O)NHCH 2 NH 2 , -C (＝O)NHCH 2 CH 2 NH 2 , -C(＝O)NHCH 2 CH 2 CH 2 NH 2 , -C(＝O)NHCH 2 SH, -C(＝O)NHCH 2 CH 2 SH, -C (＝O)NHCH 2 CH 2 CH 2 SH, -S(＝O) 2 CH 2 OH, -S(＝O) 2 CH 2 CH 2 OH, -S(＝O) 2 CH 2 CH 2 CH 2 OH , -S(＝O) 2 CH 2 NH 2 , -S(＝O) 2 CH 2 CH 2 NH 2 , -S(＝O) 2 CH 2 CH 2 CH 2 NH 2 , -S(＝O) 2 CH 2 SH, -S(=O) 2 CH 2 CH 2 SH, -S(=O) 2 CH 2 CH 2 CH 2 SH, -S(=O) 2 NHCH 2 OH, -S(=O) 2 NHCH 2 CH 2 OH, -S(＝O) 2 NHCH 2 CH 2 CH 2 OH, -S(＝O) 2 NHCH 2 NH 2 , -S(＝O) 2 NHCH 2 CH 2 NH 2 , -S( ＝O) 2 NHCH 2 CH 2 CH 2 NH 2, -S(＝O) 2 NHCH 2 SH, -S(＝O) 2 NHCH 2 CH 2 SH, -S(＝O) 2 NHCH 2 CH 2 CH 2 SH.
优选地，所述选自以下结构中的一种：
Preferably, the Choose one of the following structures:
其中，各个R9独立地选自：氢、R13、卤素、氰基、硝基、C1-C6烷基、C1-C6卤代烷基、C1-C6烷氧基；Wherein, each R 9 is independently selected from: hydrogen, R 13 , halogen, cyano, nitro, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, C 1 -C 6 alkoxy;
各个a1独立地选自：0、1、2、3、4、5、6、7、8、9，各个a2独立地选自：0、1、2、3、4、5、6、7、8，各个a3独立地选自：0、1、2、3、4、5、6、7，各个a4独立地选自：0、1、2、3、4、5、6，各个a5独立地选自：0、1、2、3、4、5。Each a1 is independently selected from: 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, each a2 is independently selected from: 0, 1, 2, 3, 4, 5, 6, 7, 8. Each a3 is independently selected from: 0, 1, 2, 3, 4, 5, 6, 7. Each a4 is independently selected from: 0, 1, 2, 3, 4, 5, 6. Each a5 is independently selected. Selected from: 0, 1, 2, 3, 4, 5.
优选地，所述R13选自：羟基、Rn、-ORn、-OC(＝O)Rn、-NRmRn、-NRm-C(＝O)Rn、-NRn-C(＝O)Rm、-NRm-S(＝O)2Rn、-NRn-S(＝O)2Rm；优选地，所述R13选自：羟基、Rn、-ORn。Preferably, the R 13 is selected from: hydroxyl, R n , -OR n , -OC(=O)R n , -NR m R n , -NR m -C(=O)R n , -NR n - C(=O)R m , -NR m -S(=O) 2 R n , -NR n -S(=O) 2 R m ; Preferably, the R 13 is selected from: hydroxyl, R n , - OR n .
优选地，所述Rn选自：羟基取代的C1-C8烷基；更优选地，Rn选自：羟基取代的C1-C4烷基；最优选地，Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH或-CH2CH2CH2CH2OH。在一些实施方案中，所述Rn选自：羟基、巯基或氨基取代的C1-C8烷基；更优选的，所述Rn选自：羟基、巯基或氨基取代的C1-C4烷基； 最优选的，Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH、-CH2CH2CH2CH2OH、-CH2NH2、-CH2CH2NH2、-CH2CH2CH2NH2、-CH2CH2CH2CH2NH2、-CH2SH、-CH2CH2SH、-CH2CH2CH2SH、-CH2CH2CH2CH2SH。Preferably, the R n is selected from: hydroxyl-substituted C 1 -C 8 alkyl; more preferably, R n is selected from: hydroxyl-substituted C 1 -C 4 alkyl; most preferably, R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH or -CH 2 CH 2 CH 2 CH 2 OH. In some embodiments, the R n is selected from: hydroxyl, mercapto or amino-substituted C 1 -C 8 alkyl; more preferably, the R n is selected from: hydroxyl, mercapto or amino substituted C 1 -C 4 alkyl; Most preferably, R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 CH 2 SH.
优选地，所述Rm选自：氢、-C1-C4烷基。Preferably, the R m is selected from: hydrogen, -C 1 -C 4 alkyl.
优选地，所述选自以下结构中的一种：
Preferably, the Choose one of the following structures:
在一些实施方案中，所述T、Z分别独立地选自：化学键、羰基、C1-C6亚烷基或C3-C10亚环烷基，所述C1-C6亚烷基、C3-C10亚环烷基任选被一个或多个R9取代。In some embodiments, the T and Z are independently selected from: chemical bond, carbonyl group, C 1 -C 6 alkylene group or C 3 -C 10 cycloalkylene group, the C 1 -C 6 alkylene group , C 3 -C 10 cycloalkylene is optionally substituted by one or more R 9 .
优选地，所述T、Z分别独立地选自：化学键、羰基、亚甲基、1,2-亚乙基、1,1-亚环丙基或2,2-亚丙基，所述亚甲基、1,2-亚乙基、1,1-亚环丙基任选被一个或多个R9取代。Preferably, the T and Z are independently selected from: chemical bonds, carbonyl groups, methylene groups, 1,2-ethylene groups, 1,1-cyclopropylene groups or 2,2-propylene groups. Methyl, 1,2-ethylene, and 1,1-cyclopropylene are optionally substituted by one or more R 9 .
优选地，所述R9选自：氢、R13、C1-C6烷基或C3-C8环烷基。在一些实施方案中，所述R9选自：氢、-R21N(R22)R22、-R21C(＝O)R22、R13、C1-C6烷基或C3-C8环烷基。Preferably, the R 9 is selected from: hydrogen, R 13 , C 1 -C 6 alkyl or C 3 -C 8 cycloalkyl. In some embodiments, the R 9 is selected from: hydrogen, -R 21 N(R 22 )R 22 , -R 21 C(=O)R 22 , R 13 , C 1 -C 6 alkyl, or C 3 -C 8 cycloalkyl.
优选地，所述R13选自：羟基、Rn、-ORn。Preferably, the R 13 is selected from: hydroxyl, R n , -OR n .
优选地，所述Rn选自：羟基取代的C1-C8烷基；更优选地，Rn选自：羟基取代的C1-C4烷基；最优选地，所述Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH或-CH2CH2CH2CH2OH。在一些实施方案中，所述Rn选自：羟基、巯基或氨基取代的C1-C8烷基；优选的，所述Rn选自：羟基、巯基或氨基取代的C1-C4烷基；更优选的，所述Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH、-CH2CH2CH2CH2OH、-CH2NH2、-CH2CH2NH2、-CH2CH2CH2NH2、-CH2CH2CH2CH2NH2、-CH2SH、-CH2CH2SH、-CH2CH2CH2SH、-CH2CH2CH2CH2SH。Preferably, the R n is selected from: hydroxyl-substituted C 1 -C 8 alkyl; more preferably, R n is selected from: hydroxyl-substituted C 1 -C 4 alkyl; most preferably, the R n is selected from From: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, or -CH 2 CH 2 CH 2 CH 2 OH. In some embodiments, the R n is selected from: hydroxyl, mercapto or amino-substituted C 1 -C 8 alkyl; preferably, the R n is selected from: hydroxyl, mercapto or amino substituted C 1 -C 4 Alkyl group; more preferably, the R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 CH 2 SH.
在一些实施方案中，所述U选自：C1-C6亚烷基、C3-C10亚环烷基、亚芳基或亚杂芳基，所述亚烷基、亚环烷基、亚芳基、亚杂芳基任选被一个或多个R9取代。 In some embodiments, the U is selected from: C 1 -C 6 alkylene, C 3 -C 10 cycloalkylene, arylene or heteroarylene, the alkylene, cycloalkylene , arylene group and heteroarylene group are optionally substituted by one or more R 9 .
优选地，所述U选自：C2-C6亚烷基、C5-C6亚环烷基、C6-C10亚芳基、5-6元单环亚杂芳基，所述亚烷基、亚环烷基、亚芳基、亚杂芳基任选被一个或多个R9取代。Preferably, the U is selected from: C 2 -C 6 alkylene, C 5 -C 6 cycloalkylene, C 6 -C 10 arylene, 5-6 membered monocyclic heteroarylene, the Alkylene, cycloalkylene, arylene, heteroarylene are optionally substituted by one or more R 9 .
优选地，所述U选自：1,2-亚乙基、1,3-亚丙基、1,4-亚丁基、1,5-亚戊基、1,6-亚己基、1,3-亚环戊基、1,3-亚环己基、1,4-亚环己基、1,2-亚苯基、1,3-亚苯基、1,4-亚苯基、2,5-亚吡啶基、2,5-亚嘧啶基、2,5-亚噻唑基或2,4-亚噁唑基，所述1,2-亚乙基、1,3-亚丙基、1,4-亚丁基、1,3-亚环戊基、1,3-亚环己基、1,4-亚环己基、1,2-亚苯基、1,3-亚苯基、1,4-亚苯基、2,5-亚吡啶基、2,5-亚嘧啶基、2,5-亚噻唑基、2,4-亚噁唑基任选被一个或多个R9取代。Preferably, the U is selected from: 1,2-ethylene, 1,3-propylene, 1,4-butylene, 1,5-pentylene, 1,6-hexylene, 1,3 -Cyclopentylene, 1,3-cyclohexylene, 1,4-cyclohexylene, 1,2-phenylene, 1,3-phenylene, 1,4-phenylene, 2,5- Pyridinyl, 2,5-pyrimidinyl, 2,5-thiazolyl or 2,4-oxazolyl, the 1,2-ethylene, 1,3-propylene, 1,4 -Butylene, 1,3-cyclopentylene, 1,3-cyclohexylene, 1,4-cyclohexylene, 1,2-phenylene, 1,3-phenylene, 1,4-phenylene Phenyl, 2,5-pyridinylene, 2,5-pyrimidinyl, 2,5-thiazolylene, and 2,4-oxazolylene are optionally substituted by one or more R 9 .
优选地，所述R9选自：氢、R13、卤素、C1-C6烷基、C1-C6卤代烷基、C1-C6烷氧基。Preferably, the R 9 is selected from: hydrogen, R 13 , halogen, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, C 1 -C 6 alkoxy.
优选地，所述R13选自：羟基、Rn、-ORn、-OC(＝O)Rn、-NRmRn、-NRm-C(＝O)Rn、-NRn-C(＝O)Rm、-NRm-S(＝O)2Rn、-NRn-S(＝O)2Rm；更优选地，所述R13选自：羟基、Rn、-ORn、-NRmRn、-NRn-C(＝O)Rm、-NRn-S(＝O)2Rm。Preferably, the R 13 is selected from: hydroxyl, R n , -OR n , -OC(=O)R n , -NR m R n , -NR m -C(=O)R n , -NR n - C(=O)R m , -NR m -S(=O) 2 R n , -NR n -S(=O) 2 R m ; more preferably, the R 13 is selected from: hydroxyl, R n , -OR n , -NR m R n , -NR n -C(=O)R m , -NR n -S(=O) 2 R m .
优选地，所述Rn选自：羟基取代的C1-C8烷基；更优选地，Rn选自：羟基取代的C1-C4烷基；最优选地，Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH或-CH2CH2CH2CH2OH。在一些实施方案中，所述Rn选自：羟基、巯基或氨基取代的C1-C8烷基；优选的，所述Rn选自：羟基、巯基或氨基取代的C1-C4烷基；更优选的，所述Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH、-CH2CH2CH2CH2OH、-CH2NH2、-CH2CH2NH2、-CH2CH2CH2NH2、-CH2CH2CH2CH2NH2、-CH2SH、-CH2CH2SH、-CH2CH2CH2SH、-CH2CH2CH2CH2SH。Preferably, the R n is selected from: hydroxyl-substituted C 1 -C 8 alkyl; more preferably, R n is selected from: hydroxyl-substituted C 1 -C 4 alkyl; most preferably, R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH or -CH 2 CH 2 CH 2 CH 2 OH. In some embodiments, the R n is selected from: hydroxyl, mercapto or amino-substituted C 1 -C 8 alkyl; preferably, the R n is selected from: hydroxyl, mercapto or amino substituted C 1 -C 4 Alkyl group; more preferably, the R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 CH 2 SH.
优选地，所述Rm选自：氢、-C1-C4烷基。Preferably, the R m is selected from: hydrogen, -C 1 -C 4 alkyl.
优选地，所述R9选自：F、Cl、Br、氰基、甲基、乙基、三氟甲基、甲氧基、乙氧基、羟基、-CH2OH、-CH2CH2OH、-CH2CH2CH2OH、-OCH2OH、-OCH2CH2OH、-OCH2CH2CH2OH、-NHCH2OH、-NHCH2CH2OH、-NHCH2CH2CH2OH、-N(CH3)CH2OH、-N(CH3)CH2CH2OH、-N(CH3)CH2CH2CH2OH、-N(COCH3)CH2OH、-N(COCH3)CH2CH2OH、-N(COCH3)CH2CH2CH2OH、-N(SO2CH3)CH2OH、-N(SO2CH3)CH2CH2OH、-N(SO2CH3)CH2CH2CH2OH。在一些实施方案中，所述R9选自：F、Cl、Br、氰基、甲基、乙基、三氟甲基、甲氧基、乙氧基、羟基、 巯基、氨基、-C(＝O)CH3、C(＝O)CH2CH3、C(＝O)CH2CH2CH3、-CH2CH2N(CH3)2、-CH2OH、-CH2CH2OH、-CH2CH2CH2OH、-CH2NH2、-CH2CH2NH2、-CH2CH2CH2NH2、-CH2SH、-CH2CH2SH、-CH2CH2CH2SH、-CH2CH2CH2CH2SH、-OCH2OH、-OCH2CH2OH、-OCH2CH2CH2OH、-OCH2NH2、-OCH2CH2NH2、-OCH2CH2CH2NH2、-OCH2SH、-OCH2CH2SH、-OCH2CH2CH2SH、-NHCH2OH、-NHCH2CH2OH、-NHCH2CH2CH2OH、-NHCH2NH2、-NHCH2CH2NH2、-NHCH2CH2CH2NH2、-NHCH2SH、-NHCH2CH2SH、-NHCH2CH2CH2SH、-N(CH3)CH2OH、-N(CH3)CH2CH2OH、-N(CH3)CH2CH2CH2OH、-N(CH3)CH2NH2、-N(CH3)CH2CH2NH2、-N(CH3)CH2CH2CH2NH2、-N(CH3)CH2SH、-N(CH3)CH2CH2SH、-N(CH3)CH2CH2CH2SH、-N(COCH3)CH2OH、-N(COCH3)CH2CH2OH、-N(COCH3)CH2CH2CH2OH、-N(COCH3)CH2NH2、-N(COCH3)CH2CH2NH2、-N(COCH3)CH2CH2CH2NH2、-N(COCH3)CH2SH、-N(COCH3)CH2CH2SH、-N(COCH3)CH2CH2CH2SH-N(SO2CH3)CH2OH、-N(SO2CH3)CH2CH2OH、-N(SO2CH3)CH2CH2CH2OH、-N(SO2CH3)CH2NH2、-N(SO2CH3)CH2CH2NH2、-N(SO2CH3)CH2CH2CH2NH2、-N(SO2CH3)CH2SH、-N(SO2CH3)CH2CH2SH、-N(SO2CH3)CH2CH2CH2SH。Preferably, the R 9 is selected from: F, Cl, Br, cyano, methyl, ethyl, trifluoromethyl, methoxy, ethoxy, hydroxyl, -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -OCH 2 OH, -OCH 2 CH 2 OH, -OCH 2 CH 2 CH 2 OH, -NHCH 2 OH, -NHCH 2 CH 2 OH, -NHCH 2 CH 2 CH 2 OH, -N(CH 3 )CH 2 OH, -N(CH 3 )CH 2 CH 2 OH, -N(CH 3 )CH 2 CH 2 CH 2 OH, -N(COCH 3 )CH 2 OH, - N(COCH 3 )CH 2 CH 2 OH, -N(COCH 3 )CH 2 CH 2 CH 2 OH, -N(SO 2 CH 3 )CH 2 OH, -N(SO 2 CH 3 )CH 2 CH 2 OH , -N(SO 2 CH 3 )CH 2 CH 2 CH 2 OH. In some embodiments, the R 9 is selected from: F, Cl, Br, cyano, methyl, ethyl, trifluoromethyl, methoxy, ethoxy, hydroxyl, Mercapto group, amino group, -C(=O)CH 3 , C(=O)CH 2 CH 3 , C(=O)CH 2 CH 2 CH 3 , -CH 2 CH 2 N(CH 3 ) 2 , -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 CH 2 SH, -OCH 2 OH, -OCH 2 CH 2 OH, -OCH 2 CH 2 CH 2 OH, -OCH 2 NH 2 , -OCH 2 CH 2 NH 2 , -OCH 2 CH 2 CH 2 NH 2 , -OCH 2 SH, -OCH 2 CH 2 SH, -OCH 2 CH 2 CH 2 SH, -NHCH 2 OH, -NHCH 2 CH 2 OH, -NHCH 2 CH 2 CH 2 OH, -NHCH 2 NH 2 , -NHCH 2 CH 2 NH 2 , -NHCH 2 CH 2 CH 2 NH 2 , -NHCH 2 SH, -NHCH 2 CH 2 SH, -NHCH 2 CH 2 CH 2 SH, -N(CH 3 )CH 2 OH, -N(CH 3 )CH 2 CH 2 OH, -N(CH 3 )CH 2 CH 2 CH 2 OH, -N(CH 3 )CH 2 NH 2 , -N(CH 3 )CH 2 CH 2 NH 2 , -N(CH 3 )CH 2 CH 2 CH 2 NH 2 , -N(CH 3 )CH 2 SH, -N(CH 3 )CH 2 CH 2 SH, -N(CH 3 )CH 2 CH 2 CH 2 SH , -N(COCH 3 )CH 2 OH, -N(COCH 3 )CH 2 CH 2 OH, -N(COCH 3 )CH 2 CH 2 CH 2 OH, -N(COCH 3 )CH 2 NH 2 , -N(COCH 3 )CH 2 CH 2 NH 2 , -N(COCH 3 )CH 2 CH 2 CH 2 NH 2 , -N(COCH 3 )CH 2 SH, -N(COCH 3 )CH 2 CH 2 SH, -N(COCH 3 )CH 2 CH 2 CH 2 SH -N(SO 2 CH 3 )CH 2 OH, -N(SO 2 CH 3 )CH 2 CH 2 OH, -N(SO 2 CH 3 )CH 2 CH 2 CH 2 OH, -N(SO 2 CH 3 )CH 2 NH 2 , -N(SO 2 CH 3 )CH 2 CH 2 NH 2 , -N( SO 2 CH 3 )CH 2 CH 2 CH 2 NH 2, -N(SO 2 CH 3 )CH 2 SH, -N(SO 2 CH 3 )CH 2 CH 2 SH, -N(SO 2 CH 3 )CH 2 CH 2 CH 2 SH.
优选地，所述U选自以下基团：
Preferably, said U is selected from the following groups:
在一些实施方案中，所述-T-U-Z-一起形成选自以下的基团：
In some embodiments, the -TUZ- taken together form a group selected from:
在一些实施方案中，所述Y选自：化学键、-NR14C(＝O)-、-C(＝O)NR14-、-C(＝O)NR14CH2-、-C(＝O)NHCHR9-、-NR14C(＝O)CH2-、-NHC(＝O)CHR9-、-O-、-OCHR9-、-OCHR9CH2-、-OCH2CHR9-、-NR2-、-NR2CH2-、-NHCHR9-。In some embodiments, the Y is selected from: chemical bond, -NR 14 C(=O)-, -C(=O)NR 14 -, -C(=O)NR 14 CH 2 -, -C(= O)NHCHR 9 -, -NR 14 C(=O)CH 2 -, -NHC(=O)CHR 9 -, -O-, -OCHR 9 -, -OCHR 9 CH 2 -, -OCH 2 CHR 9 - , -NR 2 -, -NR 2 CH 2 -, -NHCHR 9 -.
优选地，所述R9选自：氢、C1-C4烷基、R13；更优选地，所述R9选自：R13。Preferably, the R 9 is selected from: hydrogen, C 1 -C 4 alkyl, R 13 ; more preferably, the R 9 is selected from: R 13 .
优选地，所述R13选自：羟基、Rn、-ORn。Preferably, the R 13 is selected from: hydroxyl, R n , -OR n .
优选地，所述R14选自：Rn、-C(＝O)Rn。 Preferably, the R 14 is selected from: R n , -C(=O)R n .
优选地，所述R2选自：氢、R14、C1-C8烷基、C3-C10环烷基、芳基、-C(＝O)C3-C10环烷基、-C(＝O)C1-C8烷基或-S(＝O)2C1-C8烷基，所述C1-C8烷基、C3-C10环烷基、芳基以及-C(＝O)C3-C10环烷基、-C(＝O)C1-C8烷基或-S(＝O)2C1-C8烷基的环烷基或烷基部分任选被一个或多个R9取代，所述R9选自：氢、卤素、氨基、氰基、羧基、C1-C6烷基、C1-C6烷氧基或芳基；更优选地，所述R2选自：R14。其中，优选地，所述R14选自：Rn、-C(＝O)Rn、-C(＝O)ORn、-C(＝O)NRmRn、-S(＝O)2Rn、-S(＝O)2NRmRn。Preferably, the R 2 is selected from: hydrogen, R 14 , C 1 -C 8 alkyl, C 3 -C 10 cycloalkyl, aryl, -C(=O)C 3 -C 10 cycloalkyl, -C(=O)C 1 -C 8 alkyl or -S(=O) 2 C 1 -C 8 alkyl, the C 1 -C 8 alkyl, C 3 -C 10 cycloalkyl, aryl group and -C(=O)C 3 -C 10 cycloalkyl, -C(=O)C 1 -C 8 alkyl or -S(=O) 2 C 1 -C 8 alkyl cycloalkyl or alkyl The base part is optionally substituted by one or more R 9 selected from: hydrogen, halogen, amino, cyano, carboxyl, C 1 -C 6 alkyl, C 1 -C 6 alkoxy or aryl ; More preferably, the R 2 is selected from: R 14 . Wherein, preferably, the R 14 is selected from: R n , -C(=O)R n , -C(=O)OR n , -C(=O)NR m R n , -S(=O) 2 R n , -S(=O) 2 NR m R n .
优选地，所述Rn选自：羟基取代的C1-C8烷基；更优选地，所述Rn选自：羟基取代的C1-C4烷基；最优选地，所述Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH或-CH2CH2CH2CH2OH。在一些实施方案中，所述Rn选自：羟基、巯基或氨基取代的C1-C8烷基；更优选的，所述Rn选自：羟基、巯基或氨基取代的C1-C4烷基；最优选的，Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH、-CH2CH2CH2CH2OH、-CH2NH2、-CH2CH2NH2、-CH2CH2CH2NH2、-CH2CH2CH2CH2NH2、-CH2SH、-CH2CH2SH、-CH2CH2CH2SH、-CH2CH2CH2CH2SH。Preferably, the R n is selected from: hydroxyl-substituted C 1 -C 8 alkyl; more preferably, the R n is selected from: hydroxyl-substituted C 1 -C 4 alkyl; most preferably, the R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH or -CH 2 CH 2 CH 2 CH 2 OH. In some embodiments, the R n is selected from: hydroxyl, mercapto or amino-substituted C 1 -C 8 alkyl; more preferably, the R n is selected from: hydroxyl, mercapto or amino substituted C 1 -C 4 alkyl; most preferably, R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 CH 2 SH.
优选地，所述Rm选自：氢、-C1-C4烷基。Preferably, the R m is selected from: hydrogen, -C 1 -C 4 alkyl.
优选地，所述R2选自：氢、甲基、乙基、-SO2CH3、-COCH3、-CO-异丙基、-CO-环丙基、异丙基、环丙基、2-甲氧基乙基、2-氰基乙基、苯基、萘基、苄基、2-苯基乙基、1-萘甲基、2-萘乙基、-CH2OH、-CH2CH2OH、-CH2CH2CH2OH、-C(＝O)CH2OH、-C(＝O)CH2CH2OH、-C(＝O)CH2CH2CH2OH、-C(＝O)OCH2OH、-C(＝O)OCH2CH2OH、-C(＝O)OCH2CH2CH2OH、-C(＝O)NHCH2OH、-C(＝O)NHCH2CH2OH、-C(＝O)NHCH2CH2CH2OH、-S(＝O)2CH2OH、-S(＝O)2CH2CH2OH、-S(＝O)2CH2CH2CH2OH、-S(＝O)2NHCH2OH、-S(＝O)2NHCH2CH2OH、-S(＝O)2NHCH2CH2CH2OH。在一些实施方案中，所述R2进一步选自：氢、甲基、乙基、-SO2CH3、-COCH3、-CO-异丙基、-CO-环丙基、异丙基、环丙基、2-甲氧基乙基、2-氰基乙基、苯基、萘基、苄基、2-苯基乙基、1-萘甲基、2-萘乙基、-CH2OH、-CH2CH2OH、-CH2CH2CH2OH、-CH2NH2、-CH2CH2NH2、-CH2CH2CH2NH2、-CH2SH、-CH2CH2SH、-CH2CH2CH2SH、-CH2CH2CH2CH2SH、-C(＝O)CH2OH、-C(＝O)CH2CH2OH、-C(＝O)CH2CH2CH2OH、-C(＝O)CH2NH2、-C(＝O)CH2CH2NH2、-C(＝O)CH2CH2CH2NH2、-C(＝O)CH2SH、 -C(＝O)CH2CH2SH、-C(＝O)CH2CH2CH2SH、-C(＝O)OCH2OH、-C(＝O)OCH2CH2OH、-C(＝O)OCH2CH2CH2OH、-C(＝O)OCH2NH2、-C(＝O)OCH2CH2NH2、-C(＝O)OCH2CH2CH2NH2、-C(＝O)OCH2SH、-C(＝O)OCH2CH2SH、-C(＝O)OCH2CH2CH2SH、-C(＝O)NHCH2OH、-C(＝O)NHCH2CH2OH、-C(＝O)NHCH2CH2CH2OH、-C(＝O)NHCH2NH2、-C(＝O)NHCH2CH2NH2、-C(＝O)NHCH2CH2CH2NH2、-C(＝O)NHCH2SH、-C(＝O)NHCH2CH2SH、-C(＝O)NHCH2CH2CH2SH、-S(＝O)2CH2OH、-S(＝O)2CH2CH2OH、-S(＝O)2CH2CH2CH2OH、-S(＝O)2CH2NH2、-S(＝O)2CH2CH2NH2、-S(＝O)2CH2CH2CH2NH2、-S(＝O)2CH2SH、-S(＝O)2CH2CH2SH、-S(＝O)2CH2CH2CH2SH、-S(＝O)2NHCH2OH、-S(＝O)2NHCH2CH2OH、-S(＝O)2NHCH2CH2CH2OH、-S(＝O)2NHCH2NH2、-S(＝O)2NHCH2CH2NH2、-S(＝O)2NHCH2CH2CH2NH2、-S(＝O)2NHCH2SH、-S(＝O)2NHCH2CH2SH、-S(＝O)2NHCH2CH2CH2SH。Preferably, the R 2 is selected from: hydrogen, methyl, ethyl, -SO 2 CH 3 , -COCH 3 , -CO-isopropyl, -CO-cyclopropyl, isopropyl, cyclopropyl, 2-methoxyethyl, 2-cyanoethyl, phenyl, naphthyl, benzyl, 2-phenylethyl, 1-naphthylmethyl, 2-naphthylethyl, -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -C(＝O)CH 2 OH, -C(＝O)CH 2 CH 2 OH, -C(＝O)CH 2 CH 2 CH 2 OH, -C(＝O)OCH 2 OH, -C(＝O)OCH 2 CH 2 OH, -C(＝O)OCH 2 CH 2 CH 2 OH, -C(＝O)NHCH 2 OH, -C(＝ O)NHCH 2 CH 2 OH, -C(＝O)NHCH 2 CH 2 CH 2 OH, -S(＝O) 2 CH 2 OH, -S(＝O) 2 CH 2 CH 2 OH, -S(＝ O) 2 CH 2 CH 2 CH 2 OH, -S(=O) 2 NHCH 2 OH, -S(=O) 2 NHCH 2 CH 2 OH, -S(=O) 2 NHCH 2 CH 2 CH 2 OH. In some embodiments, the R 2 is further selected from: hydrogen, methyl, ethyl, -SO 2 CH 3 , -COCH 3 , -CO-isopropyl, -CO-cyclopropyl, isopropyl, Cyclopropyl, 2-methoxyethyl, 2-cyanoethyl, phenyl, naphthyl, benzyl, 2-phenylethyl, 1-naphthylmethyl, 2-naphthylethyl, -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 CH 2 SH, -C(＝O)CH 2 OH, -C(＝O)CH 2 CH 2 OH, -C(＝ O)CH 2 CH 2 CH 2 OH, -C(＝O)CH 2 NH 2 , -C(＝O)CH 2 CH 2 NH 2 , -C(＝O)CH 2 CH 2 CH 2 NH 2 , - C(=O)CH 2 SH, -C(＝O)CH 2 CH 2 SH, -C(＝O)CH 2 CH 2 CH 2 SH, -C(＝O)OCH 2 OH, -C(＝O)OCH 2 CH 2 OH, -C (＝O)OCH 2 CH 2 CH 2 OH, -C(＝O)OCH 2 NH 2 , -C(＝O)OCH 2 CH 2 NH 2 , -C(＝O)OCH 2 CH 2 CH 2 NH 2 , -C(＝O)OCH 2 SH , -C(＝O)OCH 2 CH 2 SH , -C(＝O)OCH 2 CH 2 CH 2 SH , -C(＝O)NHCH 2 OH , -C( ＝O)NHCH 2 CH 2 OH, -C(＝O)NHCH 2 CH 2 CH 2 OH, -C(＝O)NHCH 2 NH 2 , -C(＝O)NHCH 2 CH 2 NH 2 , -C( ＝O)NHCH 2 CH 2 CH 2 NH 2 , -C(＝O)NHCH 2 SH , -C(＝O)NHCH 2 CH 2 SH , -C(＝O)NHCH 2 CH 2 CH 2 SH , -S (＝O) 2 CH 2 OH, -S(＝O) 2 CH 2 CH 2 OH, -S(＝O) 2 CH 2 CH 2 CH 2 OH, -S(＝O) 2 CH 2 NH 2 , - S(＝O) 2 CH 2 CH 2 NH 2 , -S(＝O) 2 CH 2 CH 2 CH 2 NH 2 , -S(＝O) 2 CH 2 SH, -S(＝O) 2 CH 2 CH 2 SH, -S(＝O) 2 CH 2 CH 2 CH 2 SH, -S(＝O) 2 NHCH 2 OH, -S(＝O) 2 NHCH 2 CH 2 OH, -S(＝O) 2 NHCH 2 CH 2 CH 2 OH, -S(＝O) 2 NHCH 2 NH 2 , -S(＝O) 2 NHCH 2 CH 2 NH 2 , -S(＝O) 2 NHCH 2 CH 2 CH 2 NH 2, - S(=O) 2 NHCH 2 SH, -S(=O) 2 NHCH 2 CH 2 SH, -S(=O) 2 NHCH 2 CH 2 CH 2 SH.
优选地，所述R14选自：Rn、-C(＝O)Rn。Preferably, the R 14 is selected from: R n , -C(=O)R n .
优选地，所述Rn选自：羟基取代的C1-C8烷基；更优选地，Rn选自：羟基取代的C1-C4烷基；最优选地，Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH或-CH2CH2CH2CH2OH。在一些实施方案中，所述Rn选自：羟基、巯基或氨基取代的C1-C8烷基；更优选的，所述Rn选自：羟基、巯基或氨基取代的C1-C4烷基；最优选的，Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH、-CH2CH2CH2CH2OH、-CH2NH2、-CH2CH2NH2、-CH2CH2CH2NH2、-CH2CH2CH2CH2NH2、-CH2SH、-CH2CH2SH、-CH2CH2CH2SH、-CH2CH2CH2CH2SH。Preferably, the R n is selected from: hydroxyl-substituted C 1 -C 8 alkyl; more preferably, R n is selected from: hydroxyl-substituted C 1 -C 4 alkyl; most preferably, R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH or -CH 2 CH 2 CH 2 CH 2 OH. In some embodiments, the R n is selected from: hydroxyl, mercapto or amino-substituted C 1 -C 8 alkyl; more preferably, the R n is selected from: hydroxyl, mercapto or amino substituted C 1 -C 4 alkyl; most preferably, R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 CH 2 SH.
在一些实施方案中，所述X选自：-CH2-、-CH2CH2-、-C(＝O)-、-C(OH)-。In some embodiments , the X is selected from: -CH2- , -CH2CH2- , -C(=O)-, -C(OH)-.
优选地，所述X选自：-CH2-、-C(＝O)-、-CH(OH)-。Preferably, the X is selected from: -CH 2 -, -C(=O)-, -CH(OH)-.
在一些实施方案中，所述R3-R5分别独立地选自：氢、R13、卤素、氰基、氨基、C1-C6烷基、C1-C6烷氧基、C1-C6烷硫基、C1-C6烷胺基、二(C1-C6烷基)胺基、C3-C8环烷基，所述烷基、烷氧基、烷胺基、烷硫基、环烷基任选被1-3个分别选自卤素、C1-C3烷基或C1-C3烷氧基的基团取代；当R3-R5为多个的情况下，任意相邻两个可结合形成环。In some embodiments, each of R 3 to R 5 is independently selected from: hydrogen, R 13 , halogen, cyano, amino, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkylthio group, C 1 -C 6 alkylamino group, di(C 1 -C 6 alkyl)amine group, C 3 -C 8 cycloalkyl group, the alkyl group, alkoxy group, alkylamino group , alkylthio and cycloalkyl are optionally substituted by 1-3 groups respectively selected from halogen, C 1 -C 3 alkyl or C 1 -C 3 alkoxy; when R 3 -R 5 is multiple In this case, any two adjacent ones can be combined to form a ring.
优选地，所述R3选自：R13。 Preferably, said R 3 is selected from: R 13 .
优选地，所述R13选自：羟基、Rn、-ORn、-OC(＝O)Rn、-NRmRn、-NRm-C(＝O)Rn、-NRn-C(＝O)Rm、-NRm-S(＝O)2Rn、-NRn-S(＝O)2Rm；更优选地，所述R13选自：羟基、Rn、-ORn。Preferably, the R 13 is selected from: hydroxyl, R n , -OR n , -OC(=O)R n , -NR m R n , -NR m -C(=O)R n , -NR n - C(=O)R m , -NR m -S(=O) 2 R n , -NR n -S(=O) 2 R m ; more preferably, the R 13 is selected from: hydroxyl, R n , -OR n .
优选地，所述Rn选自：羟基取代的C1-C8烷基；更优选地，Rn选自：羟基取代的C1-C4烷基；最优选地，Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH或-CH2CH2CH2CH2OH。在一些实施方案中，所述Rn选自：羟基、巯基或氨基取代的C1-C8烷基；更优选的，所述Rn选自：羟基、巯基或氨基取代的C1-C4烷基；最优选的，Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH、-CH2CH2CH2CH2OH、-CH2NH2、-CH2CH2NH2、-CH2CH2CH2NH2、-CH2CH2CH2CH2NH2、-CH2SH、-CH2CH2SH、-CH2CH2CH2SH、-CH2CH2CH2CH2SH。Preferably, the R n is selected from: hydroxyl-substituted C 1 -C 8 alkyl; more preferably, R n is selected from: hydroxyl-substituted C 1 -C 4 alkyl; most preferably, R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH or -CH 2 CH 2 CH 2 CH 2 OH. In some embodiments, the R n is selected from: hydroxyl, mercapto or amino-substituted C 1 -C 8 alkyl; more preferably, the R n is selected from: hydroxyl, mercapto or amino substituted C 1 -C 4 alkyl; most preferably, R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 CH 2 SH.
优选地，Rm选自：氢、-C1-C4烷基。Preferably, R m is selected from: hydrogen, -C 1 -C 4 alkyl.
优选地，所述R4-R5分别独立地选自：氢、卤素、氰基、C1-C4烷基、C1-C4卤代烷基。Preferably, the R 4 to R 5 are each independently selected from: hydrogen, halogen, cyano, C 1 -C 4 alkyl, C 1 -C 4 haloalkyl.
在一些实施方案中，所述R6选自：氢、C1-C6烷基、C3-C8环烷基，所述烷基、环烷基任选被1-3个分别选自卤素、羟基、氰基、氨基、-C(＝O)OR23、-OC(＝O)R23、-NHC(＝O)R23、-C(＝O)NHR23或-OP(＝O)(OM)2的基团取代。In some embodiments, the R 6 is selected from: hydrogen, C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl, and the alkyl and cycloalkyl are optionally selected from 1 to 3 respectively. Halogen, hydroxyl, cyano, amino, -C(=O)OR 23 , -OC(=O)R 23 , -NHC(=O)R 23 , -C(=O)NHR 23 or -OP(=O )(OM) 2 group substitution.
在一些实施方案中，所述R6选自：氢、C1-C4烷基、C1-C4卤代烷基、-C1-C4亚烷基OC(＝O)R23、-C1-C4亚烷基C(＝O)OR23、-C1-C4亚烷基OP(＝O)(OH)2；R23选自：氢、C1-C4烷基，所述C1-C4烷基任选被1-3个分别选自羟基、氨基的基团取代。In some embodiments, the R 6 is selected from: hydrogen, C 1 -C 4 alkyl, C 1 -C 4 haloalkyl, -C 1 -C 4 alkylene OC(=O)R 23 , -C 1 -C 4 alkylene C(=O)OR 23 , -C 1 -C 4 alkylene OP(=O)(OH) 2 ; R 23 is selected from: hydrogen, C 1 -C 4 alkyl, so The C 1 -C 4 alkyl group is optionally substituted by 1 to 3 groups respectively selected from hydroxyl and amino groups.
在一些实施方案中，所述R6选自：氢、C1-C4烷基、C1-C4卤代烷基、-CH2OC(＝O)R23、-CH2CH2OC(＝O)R23、-CH2C(＝O)OR23、-CH2CH2C(＝O)OR23、-CH2OP(＝O)(OH)2、-CH2CH2OP(＝O)(OH)2；R23选自：氢、C1-C4烷基，所述C1-C4烷基任选被1-3个分别选自羟基、氨基的基团取代。In some embodiments, the R 6 is selected from: hydrogen, C 1 -C 4 alkyl, C 1 -C 4 haloalkyl, -CH 2 OC(=O)R 23 , -CH 2 CH 2 OC(= O)R 23 , -CH 2 C(=O)OR 23 , -CH 2 CH 2 C(=O)OR 23 , -CH 2 OP(=O)(OH) 2 , -CH 2 CH 2 OP(= O)(OH) 2 ; R 23 is selected from: hydrogen, C 1 -C 4 alkyl, and the C 1 -C 4 alkyl is optionally substituted by 1-3 groups respectively selected from hydroxyl and amino.
在一些实施方案中，所述R6选自：氢、C1-C4烷基、C1-C4卤代烷基、-CH2OC(＝O)C1-C4烷基、-CH2OC(＝O)CH2OH、-CH2OC(＝O)CH2NH2、-CH2OP(＝O)(OH)2。In some embodiments, the R 6 is selected from: hydrogen, C 1 -C 4 alkyl, C 1 -C 4 haloalkyl, -CH 2 OC(=O)C 1 -C 4 alkyl, -CH 2 OC(=O)CH 2 OH, -CH 2 OC(=O)CH 2 NH 2 , -CH 2 OP(=O)(OH) 2 .
在一些实施方案中，所述R6选自：氢、C1-C6烷基、C3-C8环烷基，所述烷基、环烷基任选被1-3个分别选自卤素、C1-C3烷基或C1-C3烷氧基的基团取代。In some embodiments, the R 6 is selected from: hydrogen, C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl, and the alkyl and cycloalkyl are optionally selected from 1 to 3 respectively. Group substitution of halogen, C 1 -C 3 alkyl or C 1 -C 3 alkoxy.
优选地，所述R6选自：氢、C1-C4烷基、C1-C4卤代烷基。 Preferably, the R 6 is selected from: hydrogen, C 1 -C 4 alkyl, C 1 -C 4 haloalkyl.
在一些实施方案中，R1选自：Ra(CH2)a-、Ra(CH2)aC(＝O)-，其中的“CH2”任选被一个羟基取代；In some embodiments, R 1 is selected from: R a (CH 2 ) a -, R a (CH 2 ) a C(=O)-, where “CH 2 ” is optionally substituted by a hydroxyl group;
所述Ra选自以下基团：
The R a is selected from the following groups:
所述Ra进一步优选为以下基团：
The Ra is further preferably the following group:
Q选自：-NR2-和 Q is selected from: -NR 2 -and
R2选自：C1-C8烷基、-C(＝O)ORn和-C(＝O)NRmRn；R 2 is selected from: C 1 -C 8 alkyl, -C(=O)OR n and -C(=O)NR m R n ;
Rn选自：羟基取代的C1-C8烷基；优选的，Rn选自：羟基、巯基或氨基取代的C1-C8烷基；R n is selected from: hydroxyl-substituted C 1 -C 8 alkyl; preferably, R n is selected from: hydroxyl, mercapto or amino-substituted C 1 -C 8 alkyl;
Rm选自：氢和-C1-C4烷基；R m is selected from: hydrogen and -C 1 -C 4 alkyl;
选自以下结构中的一种：
 Choose one of the following structures:
T、Z分别独立地选自：化学键、羰基、亚甲基、1,2-亚乙基、1,1-亚环丙基或2,2-亚丙基；T and Z are independently selected from: chemical bond, carbonyl group, methylene group, 1,2-ethylene group, 1,1-cyclopropylene group or 2,2-propylene group;
U选自以下基团：
U is selected from the following groups:
Y选自：-C(＝O)NH(CH2)b-和-NHC(＝O)(CH2)b-；Y is selected from: -C(=O)NH(CH 2 ) b - and -NHC(=O)(CH 2 ) b -;
R3-R6分别独立地选自：氢和C1-C6烷基；R 3 to R 6 are each independently selected from: hydrogen and C 1 -C 6 alkyl;
X为CH2；X is CH 2 ;
a选自：0、1、2、3、4或5；a is selected from: 0, 1, 2, 3, 4 or 5;
b选自：0、1、2、3、4或5；b is selected from: 0, 1, 2, 3, 4 or 5;
n选自：0、1或2；n is selected from: 0, 1 or 2;
m选自：0、1或2。m is selected from: 0, 1 or 2.
在一些实施方案中，在一些实施方案中，R1选自：Ra-、RaCH2-、RaCH2CH2-、RaCH2C(＝O)-和RaCH(OH)CH2-；In some embodiments, R1 is selected from: Ra- , RaCH2- , RaCH2CH2- , RaCH2C (=O)-, and RaCH ( OH)CH 2 -;
所述Ra选自以下基团：
The R a is selected from the following groups:
所述Ra进一步优选为以下基团：
The Ra is further preferably the following group:
Q选自：-NR2-和 Q is selected from: -NR 2 -and
R2选自：氢、-C1-C4烷基、Rn、-C(＝O)Rn、-C(＝O)ORn和-C(＝O)NRmRn； R 2 is selected from: hydrogen, -C 1 -C 4 alkyl, R n , -C(=O)R n , -C(=O)OR n and -C(=O)NR m R n ;
Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH和-CH2CH2CH2CH2OH；优选地，所述Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH、-CH2CH2CH2CH2OH、-CH2NH2、-CH2CH2NH2、-CH2CH2CH2NH2、-CH2CH2CH2CH2NH2、-CH2SH、-CH2CH2SH、-CH2CH2CH2SH、-CH2CH2CH2CH2SH；R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH and -CH 2 CH 2 CH 2 CH 2 OH; preferably, the R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 CH 2 SH;
Rm为氢；R m is hydrogen;
选自以下结构中的一种：
 Choose one of the following structures:
且通过N原子与T或R1相连；and Connected to T or R 1 through N atom;
-T-U-Z-一起形成选自以下的基团：-T-U-Z- together form a group selected from:
和C1-4亚烷基(优选亚甲基)； and C 1-4 alkylene (preferably methylene);
优选地，当Q为-NR2-时，-T-U-Z-不为C1-4亚烷基；Y选自-C(＝O)NHCH2-、-C(＝O)NH-和-NHC(＝O)-；Preferably, when Q is -NR 2 -, -TUZ- is not C 1-4 alkylene; Y is selected from -C(=O)NHCH 2 -, -C(=O)NH- and -NHC( =O)-;
R3-R6分别独立地为氢；R 3 to R 6 are each independently hydrogen;
X为CH2；X is CH 2 ;
m和n为0。m and n are 0.
在一些实施方案中，R1选自：Ra-、RaCH2-、RaCH2CH2-、RaCH2C(＝O)-和RaCH(OH)CH2-；In some embodiments, R 1 is selected from: Ra - , Ra CH 2 -, Ra CH 2 CH 2 -, Ra CH 2 C(=O)-, and Ra CH(OH)CH 2 -;
所述Ra选自以下基团：
The R a is selected from the following groups:
所述Ra进一步优选为以下基团：
The Ra is further preferably the following group:
Q选自：-NR2-和 Q is selected from: -NR 2 -and
R2选自：-C1-C4烷基、-C(＝O)ORn和-C(＝O)NRmRn； R 2 is selected from: -C 1 -C 4 alkyl, -C(=O)OR n and -C(=O)NR m R n ;
Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH、-CH2CH2CH2CH2OH、-CH2NH2、-CH2CH2NH2、-CH2CH2CH2NH2、-CH2CH2CH2CH2NH2、-CH2SH、-CH2CH2SH、-CH2CH2CH2SH、-CH2CH2CH2CH2SH；R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 CH 2 SH;
优选地，所述Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH和-CH2CH2CH2CH2OH；Preferably, the R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH and -CH 2 CH 2 CH 2 CH 2 OH;
Rm为氢；R m is hydrogen;
选自以下结构中的一种：
 Choose one of the following structures:
-T-U-Z-一起形成选自以下的基团：
-TUZ- together form a group selected from:
Y选自-C(＝O)NHCH2-和-NHC(＝O)-；Y is selected from -C(=O)NHCH 2 - and -NHC(=O)-;
R3-R6分别独立地为氢；R 3 to R 6 are each independently hydrogen;
X为CH2；X is CH 2 ;
m和n为0。m and n are 0.
在一些实施方案中，所述Ra选自以下基团：
In some embodiments, the Ra is selected from the following groups:
所述Ra进一步优选为以下基团：
The Ra is further preferably the following group:
在一些实施方案中，其中所述式III所示的结构单元为如下式III-A所示的结构单元
In some embodiments, the structural unit represented by formula III is a structural unit represented by the following formula III-A
其中各变量例如R1、R3、R4、R5、R6、Q、T、U、Z、X、Y、m和n如上所定义。The variables such as R 1 , R 3 , R 4 , R 5 , R 6 , Q, T, U, Z, X, Y, m and n are as defined above.
在一些实施方案中，D选自


In some embodiments, D is selected from


在一些实施方案中，所述抗体药物偶联物结构如式IV所示，
In some embodiments, the antibody drug conjugate structure is shown in Formula IV,
其中，Tb、S、L1、L2、Rx1、Ry1、D和q的定义如本公开中任一方案所述。Wherein, Tb, S, L 1 , L 2 , R x1 , R y1 , D and q are defined as described in any scheme of this disclosure.
在一些实施方案中，适用于本发明的ADC的抗体或其抗原结合片段为抗PSMA抗体或其抗原结合片段。In some embodiments, antibodies or antigen-binding fragments thereof suitable for use in the ADCs of the invention are anti-PSMA antibodies or antigen-binding fragments thereof.
在一些实施方案中，适用于本发明的ADC的抗体或其抗原结合片段是特异性结合PSMA的抗体部分，例如Progenics公开的PSMA抗体，或其PSMA ADC中的抗体，例如在US9242012B2或WO2021/190583A1中公开的ADC或偶联物中的PSMA抗体或其抗原结合片段。In some embodiments, an antibody or antigen-binding fragment thereof suitable for use in the ADC of the invention is an antibody portion that specifically binds PSMA, such as the PSMA antibodies disclosed by Progenics, or the antibodies in its PSMA ADC, such as those in US9242012B2 or WO2021/190583A1 PSMA antibodies or antigen-binding fragments thereof in ADCs or conjugates disclosed in.
在一些实施方案中，适用于本发明的ADC的PSMA抗体或其抗原结合片段包含已知的特异性结合PSMA的抗体，例如Progenics公开的PSMA抗体，或其PSMA ADC中的抗体部分，例如在US9242012B2或WO2021/190583A1中公开的ADC或偶联物中的抗体的1、2、3、4、5、或6个CDR。In some embodiments, PSMA antibodies or antigen-binding fragments thereof suitable for use in the ADC of the invention comprise known antibodies that specifically bind PSMA, such as the PSMA antibodies disclosed by Progenics, or antibody portions thereof in PSMA ADCs, such as those in US9242012B2 Or 1, 2, 3, 4, 5, or 6 CDRs of the antibody in the ADC or conjugate disclosed in WO2021/190583A1.
在一些实施方案中，抗原结合区包含已知的特异性结合PSMA的抗体，例如Progenics公开的PSMA抗体，或其PSMA ADC中的抗体，例如在US9242012B2或WO2021/190583A1中公开的ADC或偶联物中的抗体的1、2和3个重链可变区CDR，即HCDR1、HCDR2和HCDR3。In some embodiments, the antigen-binding region comprises an antibody known to specifically bind PSMA, such as the PSMA antibodies disclosed by Progenics, or an antibody in a PSMA ADC thereof, such as the ADC or conjugate disclosed in US9242012B2 or WO2021/190583A1 The 1, 2 and 3 heavy chain variable region CDRs of the antibody, namely HCDR1, HCDR2 and HCDR3.
在一些实施方案中，抗原结合区包含已知的特异性结合PSMA的抗体，例如Progenics公开的PSMA抗体，或其PSMA ADC中的抗体，例如在US9242012B2 或WO2021/190583A1中公开的ADC或偶联物中的抗体的1、2和3个轻链可变区CDR，即LCDR1、LCDR2和LCDR3。In some embodiments, the antigen-binding region comprises an antibody known to specifically bind PSMA, such as the PSMA antibodies disclosed by Progenics, or the antibodies in its PSMA ADC, such as those disclosed in US9242012B2 Or 1, 2 and 3 light chain variable region CDRs of the antibody in the ADC or conjugate disclosed in WO2021/190583A1, namely LCDR1, LCDR2 and LCDR3.
在一些实施方案中PSMA的抗体，例如Progenics公开的PSMA抗体，或其PSMA ADC中的抗体，例如在US9242012B2或WO2021/190583A1中公开的ADC或偶联物中的抗体的3个重链可变区CDR和3个轻链可变区CDR。In some embodiments, an antibody to PSMA, such as a PSMA antibody disclosed by Progenics, or an antibody in its PSMA ADC, such as an ADC disclosed in US9242012B2 or WO2021/190583A1, or the three heavy chain variable regions of an antibody in a conjugate CDRs and 3 light chain variable region CDRs.
在一些实施方案中，抗原结合区包含已知的特异性结合PSMA的抗体，例如Progenics公开的PSMA抗体，或其PSMA ADC中的抗体，例如在US9242012B2或WO2021/190583A1中公开的ADC或偶联物中的抗体的重链可变区，和/或轻链可变区。In some embodiments, the antigen-binding region comprises an antibody known to specifically bind PSMA, such as the PSMA antibodies disclosed by Progenics, or an antibody in a PSMA ADC thereof, such as the ADC or conjugate disclosed in US9242012B2 or WO2021/190583A1 The heavy chain variable region of the antibody, and/or the light chain variable region.
在一些实施方案中，抗原结合区包含已知的特异性结合PSMA的抗体，例如Progenics公开的PSMA抗体，或其PSMA ADC中的抗体，例如在US9242012B2或WO2021/190583A1中公开的ADC或偶联物中的抗体的重链，和/或轻链。In some embodiments, the antigen-binding region comprises an antibody known to specifically bind PSMA, such as the PSMA antibodies disclosed by Progenics, or an antibody in a PSMA ADC thereof, such as the ADC or conjugate disclosed in US9242012B2 or WO2021/190583A1 The heavy chain, and/or light chain of the antibody.
在一些实施方案中，所述PSMA抗体或其抗原结合片段为含有如下CDR的抗体或其抗原结合片段：SEQ ID No.7所示的HCDR1，SEQ ID No.8所示的HCDR2，SEQ ID No.9所示的HCDR3，SEQ ID No.10所示的LCDR1，SEQ ID No.11所示的LCDR2，SEQ ID No.12所示的LCDR3。In some embodiments, the PSMA antibody or antigen-binding fragment thereof is an antibody or antigen-binding fragment thereof containing the following CDRs: HCDR1 shown in SEQ ID No. 7, HCDR2 shown in SEQ ID No. 8, SEQ ID No. HCDR3 shown in .9, LCDR1 shown in SEQ ID No.10, LCDR2 shown in SEQ ID No.11, LCDR3 shown in SEQ ID No.12.
在一些方面中，特异性结合PSMA的抗体或其抗原结合片段包含重链可变区(VH)，其中所述VHIn some aspects, an antibody or antigen-binding fragment thereof that specifically binds PSMA comprises a heavy chain variable region (VH), wherein the VH
(i)包含与SEQ ID NO:1的氨基酸序列具有至少90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列或由所述氨基酸序列组成；或者(i) Comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 1; Consisting of the amino acid sequence; or
(ii)包含SEQ ID NO:1的氨基酸序列或由所述氨基酸序列组成；或者(ii) Comprises or consists of the amino acid sequence of SEQ ID NO:1; or
(iii)包含与SEQ ID NO：1的氨基酸序列相比具有1个或多个(优选不超过10个，更优选不超过5、4、3、2、1个)的氨基酸改变(优选氨基酸置换，更优选氨基酸保守置换)的氨基酸序列由所述氨基酸序列组成，优选地，所述氨基酸改变不发生在CDR区中。(iii) Comprises one or more (preferably no more than 10, more preferably no more than 5, 4, 3, 2, 1) amino acid changes (preferably amino acid substitutions) compared to the amino acid sequence of SEQ ID NO: 1 , more preferably amino acid conservative substitution) amino acid sequence consists of the amino acid sequence, preferably, the amino acid changes do not occur in the CDR region.
在一些方面中，特异性结合PSMA的抗体或其抗原结合片段包含轻链可变区(VL)，其中所述VL In some aspects, an antibody or antigen-binding fragment thereof that specifically binds PSMA comprises a light chain variable region (VL), wherein the VL
(i)包含与SEQ ID NO：3的氨基酸序列具有至少90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列或由所述氨基酸序列组成；或者(i) Comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 3 or Consisting of the amino acid sequence; or
(ii)包含SEQ ID NO:3的氨基酸序列或由所述氨基酸序列组成；或者(ii) Comprises or consists of the amino acid sequence of SEQ ID NO:3; or
(iii)包含与SEQ ID NO:3的氨基酸序列相比具有1个或多个(优选不超过10个，更优选不超过5、4、3、2、1个)的氨基酸改变(优选氨基酸置换，更优选氨基酸保守置换)的氨基酸序列由所述氨基酸序列组成，优选地，所述氨基酸改变不发生在CDR区中。(iii) Comprises one or more (preferably no more than 10, more preferably no more than 5, 4, 3, 2, 1) amino acid changes (preferably amino acid substitutions) compared to the amino acid sequence of SEQ ID NO:3 , more preferably amino acid conservative substitution) amino acid sequence consists of the amino acid sequence, preferably, the amino acid changes do not occur in the CDR region.
在一些优选的实施方案中，所述PSMA抗体或其抗原结合片段含有SEQ ID No.1所示的VH和SEQ ID No.3所示的VL。In some preferred embodiments, the PSMA antibody or antigen-binding fragment thereof contains the VH shown in SEQ ID No. 1 and the VL shown in SEQ ID No. 3.
在一些实施方案中，所述PSMA抗体或其抗原结合片段还包含重链恒定区和/或轻链恒定区，其中In some embodiments, the PSMA antibody or antigen-binding fragment thereof further comprises a heavy chain constant region and/or a light chain constant region, wherein
重链恒定区CHHeavy chain constant region CH
(i)包含与SEQ ID NO：2的氨基酸序列具有至少90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列或由所述氨基酸序列组成；或者(i) Comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 2 or Consisting of the amino acid sequence; or
(ii)包含SEQ ID NO:2的氨基酸序列或由所述氨基酸序列组成；或者(ii) Comprises or consists of the amino acid sequence of SEQ ID NO:2; or
(iii)包含与SEQ ID NO:2的氨基酸序列相比具有1个或多个(优选不超过10个，更优选不超过5、4、3、2、1个)的氨基酸改变(优选氨基酸置换，更优选氨基酸保守置换)的氨基酸序列由所述氨基酸序列组成；(iii) Comprises one or more (preferably no more than 10, more preferably no more than 5, 4, 3, 2, 1) amino acid changes (preferably amino acid substitutions) compared to the amino acid sequence of SEQ ID NO:2 , more preferably the amino acid sequence consisting of the amino acid sequence consisting of the amino acid conservative substitution);
和/或and / or
轻链恒定区CLLight chain constant region CL
(i)包含与SEQ ID NO：4的氨基酸序列具有至少90％、91％、92％、93％、94％、95％、96％、97％、98％或99％同一性的氨基酸序列或由所述氨基酸序列组成；或者(i) Comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 4 or Consisting of the amino acid sequence; or
(ii)包含SEQ ID NO:4的氨基酸序列或由所述氨基酸序列组成；或者(ii) Comprises or consists of the amino acid sequence of SEQ ID NO: 4; or
(iii)包含与SEQ ID NO:4的氨基酸序列相比具有1个或多个(优选不超过10个，更优选不超过5、4、3、2、1个)的氨基酸改变(优选氨基酸置换，更优选氨基酸保守置换)的氨基酸序列由所述氨基酸序列组成。 (iii) Comprises one or more (preferably no more than 10, more preferably no more than 5, 4, 3, 2, 1) amino acid changes (preferably amino acid substitutions) compared to the amino acid sequence of SEQ ID NO:4 , more preferably the amino acid sequence consists of the amino acid sequence consisting of the amino acid conservative substitution).
在一些优选的实施方案中，所述PSMA抗体或其抗原结合片段含有：SEQ ID No.1所示的VH和SEQ ID No.2所示的CH，和SEQ ID No.3所示的VL和SEQ ID No.4所示的CL。In some preferred embodiments, the PSMA antibody or antigen-binding fragment thereof contains: VH shown in SEQ ID No. 1 and CH shown in SEQ ID No. 2, and VL shown in SEQ ID No. 3 and CL shown in SEQ ID No.4.
在一些优选的实施方案中，所述PSMA抗体或其抗原结合片段含有：SEQ ID No.5所示的重链和SEQ ID No.6所示的轻链。In some preferred embodiments, the PSMA antibody or antigen-binding fragment thereof contains: the heavy chain shown in SEQ ID No. 5 and the light chain shown in SEQ ID No. 6.
在一些实施方案中，所述的抗体药物偶联物(ADC)选自以下：








In some embodiments, the antibody drug conjugate (ADC) is selected from the following:








其中，q选自1-10，例如2-8。Wherein, q is selected from 1-10, such as 2-8.
在一些实施方案中，其中所述抗体药物偶联物具有选自如下范围内的任意数值的药物与抗体比率(平均DAR)：1.0-20.0、1.0-18.0、1.0-16.0、1.0-10.0、2.0-14.0、3.0-12.0、4.0-10.0、5.0-9.0、6.0-8.0或2.0-8.0。In some embodiments, wherein the antibody drug conjugate has a drug to antibody ratio (average DAR) of any value selected from the following range: 1.0-20.0, 1.0-18.0, 1.0-16.0, 1.0-10.0, 2.0 -14.0, 3.0-12.0, 4.0-10.0, 5.0-9.0, 6.0-8.0 or 2.0-8.0.
在一些实施方案中，其中所述平均DAR为2+/-0.4、4+/-0.4、6+/-0.4或8+/-0.4内任意数值。In some embodiments, the average DAR is any value within 2+/-0.4, 4+/-0.4, 6+/-0.4, or 8+/-0.4.
在一些实施方案中，其中所述平均DAR约为2.0、4.0、6.0、8.0、10.0或12.0。In some embodiments, the average DAR is about 2.0, 4.0, 6.0, 8.0, 10.0, or 12.0.
在本公开的第二方面，本公开提供式V所示的药物连接体偶联物，
In a second aspect of the present disclosure, the present disclosure provides a drug linker conjugate represented by Formula V,
所述药物连接体偶联物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物；The stereoisomer of the drug linker conjugate, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate;
其中，L’为连接体前体，D的定义如本公开中任一方案所述。Wherein, L' is a linker precursor, and D is defined as described in any scheme of this disclosure.
在一些实施方案中，提供式VI所示的药物连接体偶联物、其立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物，
In some embodiments, a drug linker conjugate represented by Formula VI, its stereoisomer, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate is provided,
其中，in,
当L1的定义如本公开中任一方案所述，且L1不为时，Lg为和抗体反应时的离去基团；L2、L3、L4和D的定义如本公开中任一方案所述；When L 1 is defined as described in any aspect of this disclosure, and L 1 is not When , Lg is the leaving group when reacting with the antibody; L 2 , L 3 , L 4 and D are defined as described in any scheme of this disclosure;
当L1为时，Lg-L1为L2、L3、L4和D的定义如本公开中任一方案所述。When L 1 is When , Lg-L 1 is L 2 , L 3 , L 4 and D are as defined in any aspect of this disclosure.
在一些实施方案中，Lg选自卤素、砜基、三级胺盐基(Me3N+、Et3N+)、重氮盐基、-OMs、MeSO2-、CF3SO3-、对甲苯磺酰基。In some embodiments, Lg is selected from halogen, sulfone, tertiary amine (Me 3 N + , Et 3 N + ), diazonium, -OMs, MeSO 2 -, CF 3 SO 3 -, p Tosyl.
在一些实施方案中，Lg选自F、Cl、Br、MeSO2-；更优选地；Lg为MeSO2-。In some embodiments, Lg is selected from F, Cl, Br, MeSO 2 -; more preferably; Lg is MeSO 2 -.
在一些实施方案中，提供式VII所示的药物连接体偶联物、其立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物，
In some embodiments, a drug linker conjugate represented by Formula VII, its stereoisomer, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate is provided,
其中，Lg、L1、L2、Rx1、Ry1和D的定义如本公开中任一方案所述。Wherein, the definitions of Lg, L 1 , L 2 , R x1 , R y1 and D are as described in any aspect of this disclosure.
在一些实施方案中，所述的药物连接体偶联物选自以下：






In some embodiments, the drug linker conjugate is selected from the following:






在本公开的第三方面，提供如下的蛋白降解剂类生物活性化合物，
In a third aspect of the present disclosure, the following protein-degrading bioactive compounds are provided,
或所述化合物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物， or a stereoisomer of the compound, a prodrug thereof, a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable solvate thereof,
其中，R1、Q、T、U、Z、Y、X、R3、R4、R5、R6、m、n的定义如本公开中任一方案所述。Among them, the definitions of R 1 , Q, T, U, Z, Y, X, R 3 , R 4 , R 5 , R 6 , m, and n are as described in any aspect of this disclosure.
在一些实施方案中，其中所述式VIII化合物具有如下式VIII-A所示的结构
In some embodiments, the compound of Formula VIII has the structure shown in Formula VIII-A below
其中各变量例如R1、R3、R4、R5、R6、Q、T、U、Z、X、Y、m和n如上所定义。The variables such as R 1 , R 3 , R 4 , R 5 , R 6 , Q, T, U, Z, X, Y, m and n are as defined above.
在一些实施方案中，蛋白降解剂类生物活性化合物结构如下：


In some embodiments, the structure of the protein-degrading bioactive compound is as follows:


在本公开的第四方面，提供制备式I所示的抗体药物偶联物的方法，其包括：将Tb分别与式V所示的药物连接体偶联物在溶剂中进行偶联反应形成C-S键的步骤。In a fourth aspect of the present disclosure, a method for preparing an antibody-drug conjugate represented by Formula I is provided, which includes: performing a coupling reaction between Tb and a drug linker conjugate represented by Formula V in a solvent to form C-S key steps.
在一些实施方案中，所述Tb与式V所示的药物连接体偶联物的摩尔比为1:(1-20)，如1:2、1:4、1:6、1:8、1:10、1:12、1:14、1:16、1:18、1:(10-20)、1:(12-20)、1:(14-20)、1:(16-20)或1:(18-20)。In some embodiments, the molar ratio of the Tb to the drug linker conjugate represented by Formula V is 1:(1-20), such as 1:2, 1:4, 1:6, 1:8, 1:10, 1:12, 1:14, 1:16, 1:18, 1:(10-20), 1:(12-20), 1:(14-20), 1:(16-20 ) or 1:(18-20).
在一些实施方案中，所述偶联反应在水和/或有机溶剂中进行。In some embodiments, the coupling reaction is performed in water and/or organic solvents.
在一些实施方案中，所述有机溶剂选自N,N-二甲基甲酰胺、N,N-二甲基乙酰胺、二甲基亚砜、N-甲基吡咯烷酮中的一种或其任意组合。In some embodiments, the organic solvent is selected from one of N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, N-methylpyrrolidone or any of them. combination.
在一些实施方案中，所述方法进一步包括将偶联产物进行纯化的步骤。In some embodiments, the method further includes the step of purifying the coupling product.
在一些实施方案中，通过层析方法对所述偶联产物进行纯化。In some embodiments, the coupling product is purified by chromatography.
在一些实施方案中，所述层析方法包括离子交换层析、疏水层析、反相层析或亲和层析中的一种或多种。In some embodiments, the chromatography method includes one or more of ion exchange chromatography, hydrophobic chromatography, reversed phase chromatography, or affinity chromatography.
在一些实施方案中，所述方法在-20-100℃，例如0-50℃，优选室温下进行。In some embodiments, the method is carried out at -20-100°C, such as 0-50°C, preferably room temperature.
在本公开的第五方面，以V-4为例，本公开提供了药物连接体偶联物的制备方法，
In the fifth aspect of the disclosure, taking V-4 as an example, the disclosure provides a method for preparing a drug linker conjugate,
第六方面，本公开提供了药物组合物，其包含如前述的抗体药物偶联物、或所述抗体药物偶联物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物；或者如前述的蛋白降解剂类生物活性化合物、或所述化合物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物；或者如前述的药物连接体偶联物、其立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物；以及任选的一种或多种药用辅料。In a sixth aspect, the present disclosure provides a pharmaceutical composition comprising the aforementioned antibody drug conjugate, or a stereoisomer of the antibody drug conjugate, a prodrug thereof, a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable salt thereof. Pharmaceutically acceptable solvates; or the aforementioned protein degrader bioactive compounds, or stereoisomers of said compounds, their prodrugs, their pharmaceutically acceptable salts or their pharmaceutically acceptable solvents compound; or as the aforementioned drug linker conjugate, its stereoisomer, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate; and optionally one or more medicinal excipients.
第七方面，本公开提供了前述的抗体药物偶联物、或所述抗体药物偶联物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物；或者如前述的药物连接体偶联物、其立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物；或者前述的药物组合物在制备用于治疗和/或预防与细胞活动异常相关的疾病(例如癌症疾病)的药物中的用途。In the seventh aspect, the present disclosure provides the aforementioned antibody drug conjugate, or the stereoisomer of the antibody drug conjugate, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate. or the aforementioned drug linker conjugate, its stereoisomer, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate; or the aforementioned pharmaceutical composition is used in the preparation of Use in medicines for the treatment and/or prevention of diseases associated with abnormal cellular activity, such as cancer diseases.
本公开还提供了预防和/或治疗与细胞活动异常相关的疾病(例如癌症疾病)的方法，其包括向个体施用前述的抗体药物偶联物、或所述抗体药物偶联物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物；或者如前述的药物连接体偶联物、其立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物；或者前述的药物组合物。The present disclosure also provides methods for preventing and/or treating diseases related to abnormal cell activity (such as cancer diseases), which include administering to an individual the aforementioned antibody drug conjugate, or a stereoisomer of the antibody drug conjugate. body, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate; or the aforementioned drug linker conjugate, its stereoisomer, its prodrug, its pharmaceutically acceptable salt or pharmaceutically acceptable solvate thereof; or the aforementioned pharmaceutical composition.
本公开还提供了前述的抗体药物偶联物、或所述抗体药物偶联物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物；或者如前述的药物连接体偶联物、其立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物；或者前述的药物组合物，它们用于治疗和/或预防疾病，例如与细胞活动异常相关的疾病(例如癌症疾病)。The present disclosure also provides the aforementioned antibody drug conjugate, or the stereoisomer of the antibody drug conjugate, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate; or Such as the aforementioned drug linker conjugates, their stereoisomers, their prodrugs, their pharmaceutically acceptable salts or their pharmaceutically acceptable solvates; or the aforementioned pharmaceutical compositions, which are used to treat and /or prevent diseases, such as those associated with abnormal cellular activity (e.g., cancer diseases).
在一些实施方案中，所述癌症疾病为实体肿瘤或血液肿瘤。 In some embodiments, the cancer disease is a solid tumor or a hematological tumor.
在一些实施方案中，所述癌症疾病选自食管癌(例如食管腺癌和食管鳞状细胞癌)、脑瘤、肺癌(例如小细胞性肺癌和非小细胞性肺癌)、鳞状上皮细胞癌、膀胱癌、胃癌、卵巢癌、腹膜癌、胰腺癌、乳腺癌、头颈癌、子宫颈癌、子宫内膜癌、结直肠癌、肝癌、肾癌、尿路上皮癌、实体瘤、非霍奇金淋巴瘤、中枢神经系统肿瘤(例如神经胶质瘤、多形性胶质母细胞瘤、胶质瘤或肉瘤)、前列腺癌或甲状腺癌。In some embodiments, the cancer disease is selected from the group consisting of esophageal cancer (eg, esophageal adenocarcinoma and esophageal squamous cell carcinoma), brain tumors, lung cancer (eg, small cell lung cancer and non-small cell lung cancer), squamous cell carcinoma , bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, colorectal cancer, liver cancer, renal cancer, urothelial cancer, solid tumors, non-Hodge cancer Gold lymphoma, central nervous system tumors (such as glioma, glioblastoma multiforme, glioma, or sarcoma), prostate cancer, or thyroid cancer.
定义：definition:
在本公开中，除非另有说明，否则本公开中使用的科学和技术名词具有本领域技术人员所通常理解的含义。并且，本公开中所用的细胞培养、分子遗传学、核酸化学、免疫学实验室操作步骤均为相应领域内广泛使用的常规步骤。同时，为了更好地理解本公开，下面提供相关术语的定义和解释。In this disclosure, unless otherwise stated, scientific and technical terms used in this disclosure have the meanings commonly understood by those skilled in the art. Moreover, the cell culture, molecular genetics, nucleic acid chemistry, and immunology laboratory procedures used in the present disclosure are routine procedures widely used in the corresponding fields. Meanwhile, in order to better understand the present disclosure, definitions and explanations of relevant terms are provided below.
如本公开所使用，术语“蛋白降解剂”是指能够将感兴趣的蛋白通过E3连接酶诱导并进行泛素化，然后通过蛋白酶体进行降解的一类化合物，优选小分子化合物。As used in this disclosure, the term "protein degrader" refers to a class of compounds, preferably small molecule compounds, that can induce and ubiquitylate a protein of interest through E3 ligases and then degrade it through the proteasome.
术语“Myc蛋白降解剂”是指具有降解Myc家族蛋白的能力的蛋白降解剂，例如对c-Myc蛋白具有降解能力的蛋白降解剂、对n-Myc蛋白具有降解能力的蛋白降解剂。The term "Myc protein degrading agent" refers to a protein degrading agent that has the ability to degrade Myc family proteins, such as a protein degrading agent that has the ability to degrade c-Myc protein and a protein degrading agent that has the ability to degrade n-Myc protein.
术语“c-Myc蛋白降解剂”是指具有降解c-Myc蛋白的能力的蛋白降解剂。The term "c-Myc protein degrading agent" refers to a protein degrading agent that has the ability to degrade c-Myc protein.
如本公开所使用，术语“蛋白降解靶向嵌合体(PROTAC)”通常包括三个部分：一个E3泛素连接酶配体和一个靶蛋白配体，两个活性配体通过特殊设计的Linker结构连接在一起，通过招募E3泛素连接酶于靶蛋白上，泛素化靶蛋白进而将靶蛋白降解。As used in this disclosure, the term "protein degradation targeting chimera (PROTAC)" generally includes three parts: an E3 ubiquitin ligase ligand and a target protein ligand, two active ligands through a specially designed Linker structure Linked together, by recruiting E3 ubiquitin ligase to the target protein, it ubiquitinates the target protein and then degrades the target protein.
如本公开所使用，术语“分子胶降解剂(MGD)”是指一类可诱导E3泛素连接酶与靶蛋白之间新型相互作用，从而导致靶蛋白降解的小分子。As used in this disclosure, the term "molecular glue degrader (MGD)" refers to a class of small molecules that can induce novel interactions between E3 ubiquitin ligases and target proteins, thereby leading to target protein degradation.
如本公开所使用，术语“药学上可接受的盐”表示指保持本发明的抗体药物偶联物或者药物连接体偶联物的生物学效应和性能的盐，并且该盐在生物学上或其它方面不是不被期望的。本发明的偶联物(包括抗体药物偶联物和药物连接体偶联物)可以以它们的药学上可接受的盐形式存在，包括酸加成盐和碱加成盐。在本发明中，药学上可接受的酸加成盐表示本发明中的偶联物与有机或无机酸形成 的盐，有机或无机酸包括但不限于盐酸、硫酸、氢溴酸、氢碘酸、磷酸、硝酸、高氯酸、乙酸、草酸、马来酸、富马酸、酒石酸、苯磺酸、甲磺酸、水杨酸、琥珀酸、柠檬酸、乳酸、丙酸、苯甲酸、对甲苯磺酸、苹果酸等。药学上可接受的碱加成盐表示本发明中的偶联物与有机或无机碱所形成的盐，包括但不限于碱金属盐，例如锂、钠或钾盐；碱土金属盐，例如钙或镁盐；有机碱盐，例如通过与含N基团的有机碱形成的铵盐。As used in this disclosure, the term "pharmaceutically acceptable salt" means a salt that retains the biological effects and properties of the antibody-drug conjugate or drug-linker conjugate of the invention, and the salt is biologically or Other aspects are not undesirable. The conjugates of the present invention (including antibody-drug conjugates and drug-linker conjugates) may exist in the form of their pharmaceutically acceptable salts, including acid addition salts and base addition salts. In the present invention, a pharmaceutically acceptable acid addition salt means that the conjugate in the present invention is formed with an organic or inorganic acid. Salts, organic or inorganic acids include but are not limited to hydrochloric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, nitric acid, perchloric acid, acetic acid, oxalic acid, maleic acid, fumaric acid, tartaric acid, benzenesulfonic acid, formazan Sulfonic acid, salicylic acid, succinic acid, citric acid, lactic acid, propionic acid, benzoic acid, p-toluenesulfonic acid, malic acid, etc. Pharmaceutically acceptable base addition salts refer to salts formed by the conjugate in the present invention and organic or inorganic bases, including but not limited to alkali metal salts, such as lithium, sodium or potassium salts; alkaline earth metal salts, such as calcium or Magnesium salts; organic base salts, such as ammonium salts formed with organic bases containing N groups.
药学上可接受的盐可使用本领域熟知的标准程序获得，例如，通过将足量的碱性化合物和提供药学上可以接受的阴离子的合适的酸反应。Pharmaceutically acceptable salts can be obtained using standard procedures well known in the art, for example, by reacting a sufficient amount of a basic compound with a suitable acid that provides a pharmaceutically acceptable anion.
如本公开所使用，术语“立体异构体”表示由于至少一个不对称中心形成的异构体。在具有一个或多个(例如一个、两个、三个或四个)不对称中心的化合物中，其可产生外消旋混合物、单一对映异构体、非对映异构体混合物和单独的非对映异构体。特定个别分子也可以几何异构体(顺式/反式)存在。类似地，本发明的化合物可以两种或更多种处于快速平衡的结构不同的形式的混合物(通常称作互变异构体)存在。互变异构体的代表性实例包括酮-烯醇互变异构体、苯酚-酮互变异构体、亚硝基-肟互变异构体、亚胺-烯胺互变异构体等。要理解，本公开的范围涵盖所有这样的以任意比例(例如60％、65％、70％、75％、80％、85％、90％、95％、96％、97％、98％、99％)的异构体或其混合物。As used in this disclosure, the term "stereoisomer" means an isomer formed due to at least one asymmetric center. In compounds with one or more (e.g., one, two, three or four) asymmetric centers, they can give rise to racemic mixtures, single enantiomers, diastereomeric mixtures and individual of diastereomers. Certain individual molecules may also exist as geometric isomers (cis/trans). Similarly, compounds of the present invention may exist as mixtures of two or more structurally distinct forms in rapid equilibrium (often referred to as tautomers). Representative examples of tautomers include keto-enol tautomers, phenol-ketone tautomers, nitroso-oxime tautomers, and imine-enamine tautomers. wait. It is to be understood that the scope of the present disclosure encompasses all such products in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99 %) isomers or mixtures thereof.
本公开中可使用实线(——)、实楔形或虚楔形描绘本发明的化合物的碳-碳键。使用实线以描绘键连至不对称碳原子的键欲表明，包括该碳原子处的所有可能的立体异构体(例如，特定的对映异构体、外消旋混合物等)。使用实或虚楔形以描绘键连至不对称碳原子的键欲表明，存在所示的立体异构体。当存在于外消旋混合物中时，使用实及虚楔形以定义相对立体化学，而非绝对立体化学。除非另外指明，否则本发明的化合物意欲可以立体异构体(其包括顺式及反式异构体、光学异构体(例如R及S对映异构体)、非对映异构体、几何异构体、旋转异构体、构象异构体、阻转异构体及其混合物)的形式存在。本发明的化合物可表现一种以上类型的异构现象，且由其混合物(例如外消旋混合物及非对映异构体对)组成。The carbon-carbon bonds of the compounds of the invention may be depicted in this disclosure using solid lines (—), solid wedges, or dashed wedges. The use of a solid line to depict a bond to an asymmetric carbon atom is intended to indicate that all possible stereoisomers at that carbon atom are included (eg, a specific enantiomer, a racemic mixture, etc.). The use of solid or imaginary wedges to depict bonds to asymmetric carbon atoms is intended to demonstrate that the stereoisomers shown exist. When present in a racemic mixture, solid and imaginary wedges are used to define relative stereochemistry rather than absolute stereochemistry. Unless otherwise specified, the compounds of the present invention are intended to exist as stereoisomers (which includes cis and trans isomers, optical isomers (e.g., R and S enantiomers), diastereomers, They exist in the form of geometric isomers, rotamers, conformational isomers, atropisomers and mixtures thereof). The compounds of the present invention may exhibit more than one type of isomerism and consist of mixtures thereof (eg, racemic mixtures and pairs of diastereoisomers).
本公开还包括所有药学上可接受的同位素化合物，其与本发明的化合物相同，除了一个或多个原子被具有相同原子序数但原子质量或质量数不同于在自然界中占优势的原子质量或质量数的原子替代。适合包含入本公开的化合物中的同位 素的实例包括(但不限于)氢的同位素(例如2H、3H)；碳的同位素(例如11C、13C及14C)；氯的同位素(例如36Cl)；氟的同位素(例如18F)；碘的同位素(例如123I及125I)；氮的同位素(例如13N及15N)；氧的同位素(例如15O、17O及18O)；磷的同位素(例如32P)；及硫的同位素(例如35S)。The present disclosure also includes all pharmaceutically acceptable isotopic compounds that are identical to the compounds of the invention except that one or more atoms are assigned the same atomic number but an atomic mass or mass number that is different from the atomic mass or mass that predominates in nature Atomic substitution of numbers. Isotopes suitable for inclusion in the compounds of the present disclosure Examples of isotopes include, but are not limited to, isotopes of hydrogen (e.g. 2 H, 3 H); isotopes of carbon (e.g. 11 C, 13 C and 14 C); isotopes of chlorine (e.g. 36 Cl); isotopes of fluorine (e.g. 18 F); Iodine isotopes (such as 123 I and 125 I); Nitrogen isotopes (such as 13 N and 15 N); Oxygen isotopes (such as 15 O, 17 O and 18 O); Phosphorus isotopes (such as 32 P ); and isotopes of sulfur (e.g. 35 S).
本公开的化合物可以溶剂合物(优选水合物)的形式存在，其中本公开的化合物包含作为所述化合物晶格的结构要素的极性溶剂，特别是例如水、甲醇或乙醇。极性溶剂特别是水的量可以化学计量比或非化学计量比存在。The compounds of the present disclosure may exist in the form of solvates, preferably hydrates, wherein the compounds of the present disclosure comprise a polar solvent as a structural element of the crystal lattice of the compound, in particular such as water, methanol or ethanol. The amount of polar solvent, especially water, may be present in stoichiometric or non-stoichiometric ratios.
在本发明的范围内还包括本发明的化合物的代谢物，即在给药本发明的化合物时体内形成的物质。这样的产物可由例如被给药的化合物的氧化、还原、水解、酰胺化、脱酰胺化、酯化、脱脂化、酶解等产生。因此，本发明包括本发明的化合物的代谢物，包括通过使本发明的化合物与哺乳动物接触足以产生其代谢产物的时间的方法制得的化合物。Also included within the scope of the invention are metabolites of the compounds of the invention, ie substances formed in the body upon administration of the compounds of the invention. Such products may result, for example, from oxidation, reduction, hydrolysis, amidation, deamidation, esterification, delipidation, enzymatic hydrolysis, etc. of the administered compound. Accordingly, the invention includes metabolites of the compounds of the invention, including compounds prepared by contacting a compound of the invention with a mammal for a time sufficient to produce metabolites thereof.
本公开在其范围内进一步包括本发明的化合物的前药。通常这样的前药会是所述化合物的官能团衍生物，其易于在体内转化成期望的治疗活性化合物。因此，在这些情况中，用于本公开的治疗方法的术语“给药”应包括用所要求保护的化合物中的一种或多种的前药形式来治疗各种疾病或病症，但是在向个体给药后所述前药形式在体内转化成上述化合物。例如，在“Design of Prodrug”,ed.H.Bundgaard,Elsevier,1985中，描述了选择和制备适合的前药衍生物的常规方法。The present disclosure further includes within its scope prodrugs of the compounds of the invention. Typically such prodrugs will be functional group derivatives of the compound that are readily converted in vivo to the desired therapeutically active compound. Therefore, in these instances, the term "administering" as used in the treatment methods of the present disclosure shall include the treatment of various diseases or conditions with prodrug forms of one or more of the claimed compounds, but in The prodrug form is converted in vivo to the compound described above upon administration to an individual. For example, in "Design of Prodrug", ed. H. Bundgaard, Elsevier, 1985, general methods for selecting and preparing suitable prodrug derivatives are described.
本公开中，所述药用辅料是指生产药品和调配处方时，使用的赋形剂和附加剂，是指除活性成分外，在安全性方面已进行了合理的评估，并且包含在药物制剂中的物质。药用辅料除了赋型、充当载体、提高稳定性外，还具有增溶、助溶、缓控释等重要功能，是可能会影响到药品的质量、安全性和有效性的重要成分。根据其来源可分为天然物、半合成物和全合成物。根据其作用与用途可分为：溶剂、抛射剂、增溶剂、助溶剂、乳化剂、着色剂、黏合剂、崩解剂、填充剂、润滑剂、湿润剂、渗透压调节剂、稳定剂、助流剂、矫味剂、防腐剂、助悬剂、包衣材料、芳香剂、抗黏着剂、抗氧剂、螯合剂、渗透促进剂、pH调节剂、缓冲剂、增塑剂、表面活性剂、发泡剂、消泡剂、增稠剂、包合剂、保湿剂、吸收剂、稀释剂、絮凝剂与反絮凝剂、助滤剂、释放阻滞剂等；根据其给药途径可分为口 服、注射、黏膜、经皮或局部给药、经鼻或口腔吸入给药和眼部给药等。同一药用辅料可用于不同给药途径的药物制剂，且有不同的作用和用途。In this disclosure, the pharmaceutical excipients refer to the excipients and additives used in the production of drugs and preparation of prescriptions. They refer to excipients and additives that have been reasonably evaluated in terms of safety in addition to active ingredients and are included in pharmaceutical preparations. substance in. In addition to excipients, serving as carriers, and improving stability, pharmaceutical excipients also have important functions such as solubilization, dissolution, and sustained and controlled release. They are important ingredients that may affect the quality, safety, and effectiveness of drugs. According to their origin, they can be divided into natural products, semi-synthetic products and fully synthetic products. According to their functions and uses, they can be divided into: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, Glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesive agents, antioxidants, chelating agents, penetration enhancers, pH regulators, buffers, plasticizers, surface active agents Agents, foaming agents, defoaming agents, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants and deflocculants, filter aids, release retardants, etc.; can be divided according to their route of administration for mouth Oral administration, injection, mucous membrane, transdermal or topical administration, nasal or oral inhalation administration and ocular administration, etc. The same pharmaceutical excipients can be used in pharmaceutical preparations with different administration routes and have different effects and uses.
所述药物组合物可根据给药途径制成各种适宜的剂型。例如片剂、胶囊剂、颗粒剂、口服溶液剂、口服混悬剂、口服乳剂、散剂、酊剂、糖浆剂、注射剂、栓剂、软膏剂、乳膏剂、糊剂、眼用制剂、丸剂、植入剂、气雾剂、粉雾剂、喷雾剂等。其中，所述的药物组合物或适宜的剂型可以含有0.01mg至1000mg的本公开的化合物(包括偶联物)或其药学上可接受的盐，适宜含有0.1mg至800mg，优选含有0.5-500mg，优选含有0.5至350mg，特别优选1-250mg。The pharmaceutical composition can be prepared into various suitable dosage forms according to the route of administration. For example, tablets, capsules, granules, oral solutions, oral suspensions, oral emulsions, powders, tinctures, syrups, injections, suppositories, ointments, creams, pastes, ophthalmic preparations, pills, implants agents, aerosols, powder mist, sprays, etc. Wherein, the pharmaceutical composition or suitable dosage form may contain 0.01 mg to 1000 mg of the compound of the present disclosure (including conjugates) or a pharmaceutically acceptable salt thereof, suitably 0.1 mg to 800 mg, preferably 0.5-500 mg. , preferably contains 0.5 to 350 mg, particularly preferably 1 to 250 mg.
所述药物组合物可以注射剂形式用药，包括注射液、注射用无菌粉末与注射用浓溶液。其中，可使用的载体和溶剂包括水、林格氏溶液和等渗氯化钠溶液。另外，灭菌的非挥发油也可用作溶剂或悬浮介质，如单甘油酯或二甘油酯。所述药物组合物可以输液形式用药。The pharmaceutical composition can be administered in the form of injection, including injection liquid, sterile powder for injection and concentrated solution for injection. Among them, carriers and solvents that can be used include water, Ringer's solution and isotonic sodium chloride solution. Alternatively, sterile fixed oils may be used as solvents or suspending media, such as mono- or diglycerides. The pharmaceutical composition may be administered in the form of an infusion.
本公开使用的术语“治疗”一般是指获得需要的药理和/或生理效应。该效应根据完全或部分地预防疾病或其症状，可以是预防性的；和/或根据部分或完全稳定或治愈疾病和/或由于疾病产生的副作用，可以是治疗性的。本公开使用的“治疗”涵盖了对患者疾病的任何治疗，包括：(a)预防易感染疾病或症状但还没诊断出患病的患者所发生的疾病或症状；(b)抑制疾病的症状，即阻止其发展；或(c)缓解疾病的症状，即，导致疾病或症状退化。The term "treatment" as used in this disclosure generally refers to obtaining a desired pharmacological and/or physiological effect. The effect may be prophylactic in terms of completely or partially preventing the disease or its symptoms; and/or may be therapeutic in terms of partially or completely stabilizing or curing the disease and/or side effects due to the disease. As used in this disclosure, "treatment" encompasses any treatment of a disease in a patient, including: (a) preventing the disease or symptoms in a patient who is susceptible to the disease or condition but has not yet been diagnosed with the disease; (b) suppressing the symptoms of the disease , i.e., arresting its progression; or (c) alleviating the symptoms of a disease, i.e., causing regression of the disease or symptoms.
在本公开中，术语“个体”包括人或非人动物。示例性人个体包括患有疾病(例如本公开所述的疾病)的人个体(称为患者)或正常个体。本公开中术语“非人动物”包括所有脊椎动物，例如非哺乳动物(例如鸟类、两栖动物、爬行动物)和哺乳动物，例如非人灵长类、家畜和/或驯化动物(例如绵羊、犬、猫、奶牛、猪等)。In this disclosure, the term "individual" includes humans or non-human animals. Exemplary human subjects include human subjects (referred to as patients) suffering from a disease, such as those described in this disclosure, or normal subjects. The term "non-human animals" in this disclosure includes all vertebrate animals, such as non-mammals (e.g., birds, amphibians, reptiles) and mammals, such as non-human primates, domestic animals, and/or domesticated animals (e.g., sheep, dogs, cats, cows, pigs, etc.).
本公开中，术语“有效剂量”指被给药后会在一定程度上缓解所治疗病症的一种或多种症状的抗体药物偶联物、药物连接体偶联物、化合物或组合物的量。In this disclosure, the term "effective dose" refers to an amount of an antibody-drug conjugate, drug-linker conjugate, compound or composition that when administered will alleviate to a certain extent one or more symptoms of the condition being treated. .
本公开中，术语“抗体药物偶联物”“ADC”是指生物活性化合物片段(药物分子)与抗体或其抗原结合片段部分连接得到的物质。在本公开的部分实施方案中，生物活性化合物片段与靶向部分通过连接体相连。所述连接体在特定环境(例如胞内低pH值环境)中或特定作用(例如溶酶体蛋白酶的作用)下能够断裂，从而使生物活性化合物(例如c-Myc蛋白降解剂)片段与靶向部分或抗体或其抗原结合片段分离。在本公开的部分实施方案中，所述连接体包含可切割或不可切割 的单元，例如肽或二硫键。在本公开的部分实施方案中，生物活性化合物片段与靶向部分或抗体或其抗原结合片段直接通过共价键相连，所述共价键在特定环境或作用下能够断裂，从而使生物活性化合物片段与抗体或其抗原结合片段部分分离。In the present disclosure, the terms "antibody drug conjugate" and "ADC" refer to a substance obtained by connecting a biologically active compound fragment (drug molecule) to an antibody or its antigen-binding fragment portion. In some embodiments of the present disclosure, the bioactive compound fragment is linked to the targeting moiety via a linker. The linker can be cleaved in a specific environment (such as an intracellular low pH environment) or under a specific action (such as the action of a lysosomal protease), thereby allowing the bioactive compound (such as a c-Myc protein degrader) fragment to interact with the target. Separation into parts or antibodies or antigen-binding fragments thereof. In some embodiments of the present disclosure, the linker comprises a cleavable or non-cleavable units such as peptides or disulfide bonds. In some embodiments of the present disclosure, the bioactive compound fragment is directly connected to the targeting moiety or antibody or antigen-binding fragment thereof by a covalent bond, which can be broken under specific circumstances or effects, thereby rendering the bioactive compound The fragment is partially separated from the antibody or antigen-binding fragment thereof.
本公开中，对于“L的1位通过S原子与Tb相连”，本领域技术人员可以理解的是，L的1位是与打开二硫键(例如，通过还原剂TCEP还原二硫键可以打开二硫键，生成巯基-SH)后的Tb(如抗体)自身所含有巯基进行连接，也就是说，L与Tb之间的-S-并非另外再外接的硫原子。例如，其中，-S-并非另外外接的硫原子，而是打开双硫键后的Tb自身所含有巯基与L，例如的1位进行连接后形成的-S-。In the present disclosure, for "the 1-position of L is connected to Tb through an S atom", those skilled in the art can understand that the 1-position of L is responsible for opening the disulfide bond (for example, the disulfide bond can be opened by reducing the disulfide bond through the reducing agent TCEP). The disulfide bond, which generates the sulfhydryl group -SH), is connected to the sulfhydryl group contained in Tb (such as an antibody) itself. In other words, the -S- between L and Tb is not an additional external sulfur atom. For example, -S- is not another external sulfur atom, but the thiol group and L contained in Tb itself after opening the disulfide bond, for example 1 bits are connected to form -S-.
本公开中，表示连接部分的连接点。In this disclosure, Indicates the connection points of connected parts.
本公开中，术语“未被取代”是指所涉及的基团未被取代基取代，应当理解上述的所涉及的基团将包含适当数量的氢原子以符合价键规则。In this disclosure, the term "unsubstituted" means that the referenced group is not substituted with a substituent, it being understood that the referenced group will contain an appropriate number of hydrogen atoms to comply with valence rules.
本公开中，术语“被…取代”指分子任一处的碳(或氮)位置处独立地存在一个或多个取代基，优选1-5个(例如1、2、3、4或5个)取代基，最优选1-3个取代基，取代基可以为：羟基、巯基、羧基、氰基、硝基、卤素(优选地，1、2或3个卤素，尤其在烷基上，尤其甲基，例如三氟甲基)、烷基(优选C1-C10，更优选地，C1-C6)、卤代烷基、烯基、炔基、环烷基、芳基(尤其苯基)、杂芳基、杂环基、烷氧基(优选C1-C6烷氧基)、芳氧基(优选苯氧基)、硫醚(C1-C6烷硫基或芳硫基如苯硫基)、酰基(优选C1-C6酰基、芳甲酰基如苯甲酰基)、C1-C6烷胺基、二(C1-C6烷基)胺基、被一或两个C1-C6烷基取代的胺基酰基(包括任选地被一个或两个C1-C6烷基取代的胺甲酰基)、C1-C6酯基等。应当理解，当术语“被取代”已经指定或列举了具体取代基时，是指被所述取代基取代。In the present disclosure, the term "substituted by" refers to the independent presence of one or more substituents at any carbon (or nitrogen) position of the molecule, preferably 1-5 (such as 1, 2, 3, 4 or 5 ) substituent, most preferably 1-3 substituents, the substituent may be: hydroxyl, mercapto, carboxyl, cyano, nitro, halogen (preferably, 1, 2 or 3 halogen, especially on the alkyl group, especially on the alkyl group Methyl, for example trifluoromethyl), alkyl (preferably C 1 -C 10 , more preferably C 1 -C 6 ), haloalkyl, alkenyl, alkynyl, cycloalkyl, aryl (especially phenyl ), heteroaryl, heterocyclyl, alkoxy (preferably C 1 -C 6 alkoxy), aryloxy (preferably phenoxy), thioether (C 1 -C 6 alkylthio or arylthio) Such as phenylthio), acyl (preferably C 1 -C 6 acyl, aroyl such as benzoyl), C 1 -C 6 alkylamino, di(C 1 -C 6 alkyl)amine, by one or Two C 1 -C 6 alkyl substituted aminoacyl groups (including carbamoyl groups optionally substituted by one or two C 1 -C 6 alkyl groups), C 1 -C 6 ester groups, etc. It will be understood that when the term "substituted" has designated or enumerated a specific substituent, it means substituted by said substituent.
本公开中，术语“任选地被取代”或类似表述是指分子或基团等可以被取代，也可以是不被取代的。对于“被取代”的情况，可参见上面的定义。In this disclosure, the term "optionally substituted" or similar expressions means that the molecule or group, etc. may be substituted or unsubstituted. For the case of "replaced", see the definition above.
本公开中，术语“烷基”是指可以任选地被取代的直链、支链完全饱和的烃基，优选C1-C10，更优选C1-C8、C1-C6，或者C1-C4烷基。烷基实例是甲基、乙基、 丙基、异丙基、正丁基、仲丁基、异丁基、叔丁基、正戊基、正己基、正庚基、正辛基、正壬基或正癸基等等。In the present disclosure, the term "alkyl" refers to a straight-chain, branched-chain fully saturated hydrocarbon group that may be optionally substituted, preferably C 1 -C 10 , more preferably C 1 -C 8 , C 1 -C 6 , or C 1 -C 4 alkyl. Examples of alkyl groups are methyl, ethyl, Propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl or n-decyl, etc.
本公开中，术语“亚烷基”是指烷基进一步去掉一个氢原子形成的二价基团。In this disclosure, the term "alkylene" refers to a divalent group formed by further removing one hydrogen atom from an alkyl group.
本公开中，术语“环烷基”是指可以任选地被取代的环状烷基，优选C3-C20，更优选C3-C15，或者C3-C8环烷基。环烷基实例是环丙基、环丁基、环戊基、环己基、环己基或环庚基等等。In the present disclosure, the term "cycloalkyl" refers to a cyclic alkyl group that may be optionally substituted, preferably C 3 -C 20 , more preferably C 3 -C 15 , or C 3 -C 8 cycloalkyl. Examples of cycloalkyl groups are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexyl or cycloheptyl, among others.
本公开中，术语“亚环烷基”是指环烷基进一步去掉一个氢原子形成的二价基团。In the present disclosure, the term "cycloalkylene" refers to a divalent group formed by further removing one hydrogen atom from a cycloalkyl group.
本公开中，术语“桥环基”是指由可以任选地被取代的两个或两个以上环状结构彼此共用两个非相邻碳原子所形成的环状结构，优选C5-C15，更优选C6-C12，或者C7-C10桥环基。桥环基的实例是双环[1.1.1]戊烷基、降冰片烷基、金刚烷基等。In the present disclosure, the term "bridged cyclic group" refers to a cyclic structure formed by two or more cyclic structures that may be optionally substituted and share two non-adjacent carbon atoms with each other, preferably C 5 -C 15 , more preferably C 6 -C 12 , or C 7 -C 10 bridged ring group. Examples of bridged cyclic groups are bicyclo[1.1.1]pentyl, norbornyl, adamantyl and the like.
本公开中，术语“烯基”是指含有至少一个C＝C键的直链、支链或环状C2-C10(优选C2-C8)烃基。In the present disclosure, the term "alkenyl" refers to a linear, branched or cyclic C 2 -C 10 (preferably C 2 -C 8 ) hydrocarbon group containing at least one C=C bond.
本公开中，术语“炔基”是指含有至少一个C≡C键的直链、支链或环状C2-C10(优选C2-C8)烃基。In the present disclosure, the term "alkynyl" refers to a linear, branched or cyclic C 2 -C 10 (preferably C 2 -C 8 ) hydrocarbon group containing at least one C≡C bond.
本公开中，术语“芳基”是指可以任选地被取代的具有单环(例如苯基)或稠环(例如萘基、蒽基、菲基、芴基等)的C6-C16芳香烃基团，优选C6-C10芳香烃基团。In the present disclosure, the term "aryl" refers to C 6 -C 16 having a single ring (such as phenyl) or a condensed ring (such as naphthyl, anthracenyl, phenanthrenyl, fluorenyl, etc.) that may be optionally substituted. Aromatic hydrocarbon groups, preferably C 6 -C 10 aromatic hydrocarbon groups.
本公开中，术语“亚芳基”是指芳基进一步去掉一个氢原子形成的二价基团。In the present disclosure, the term "arylene" refers to a divalent group formed by further removing one hydrogen atom from an aryl group.
本公开中，术语“杂芳基”是指可以任选地被取代的含有一个或多个(例如1、2、3或4个)选自N、O、S或P的杂原子的5-16元芳香族基团，优选5-10元芳香族基团，更优选5-6元芳香族基团。杂芳基的实例包括咪唑基、吡唑基、三唑基、四唑基、吡咯基、呋喃基、噻吩基、噁唑基、异噁唑基、噻唑基、异噻唑基、吡啶基、吡嗪基、嘧啶基、哒嗪基、吲哚基、氮杂吲哚基(例如7-氮杂吲哚基)、苯并咪唑基、苯并吡唑基、苯并呋喃基、苯并噻吩基、苯并噻唑基、二苯并呋喃基、二苯并噻吩基、喹啉基、异喹啉基、萘啶基、咔唑基、氮杂咔唑基(例如1-氮杂咔唑基、2-氮杂咔唑基、1,8-二氮杂咔唑基)、吲哚嗪基、氮杂吲哚嗪基、吩噁嗪基、吩噻嗪基等等。In this disclosure, the term "heteroaryl" refers to an optionally substituted 5- containing one or more (eg, 1, 2, 3, or 4) heteroatoms selected from N, O, S, or P. A 16-membered aromatic group is preferably a 5- to 10-membered aromatic group, and more preferably a 5- to 6-membered aromatic group. Examples of heteroaryl groups include imidazolyl, pyrazolyl, triazolyl, tetrazolyl, pyrrolyl, furyl, thienyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyridyl Azinyl, pyrimidinyl, pyridazinyl, indolyl, azaindolyl (such as 7-azaindolyl), benzimidazolyl, benzopyrazolyl, benzofuranyl, benzothienyl , benzothiazolyl, dibenzofuranyl, dibenzothienyl, quinolyl, isoquinolinyl, naphthyridinyl, carbazolyl, azacarbazolyl (such as 1-azacarbazolyl, 2-azacarbazolyl, 1,8-diazacarbazolyl), indolazinyl, azaindolizinyl, phenoxazinyl, phenothiazinyl, etc.
本公开中，术语“亚杂芳基”是指杂芳基进一步去掉一个氢原子形成的二价基团。 In the present disclosure, the term “heteroaryl” refers to a bivalent group formed by further removing one hydrogen atom from a heteroaryl group.
本公开中，术语“杂环基”是指可以任选地被取代的含有一个或多个(例如1、2、3或4个)选自N、O、S、SO、SO2或P的杂原子的部分不饱和或完全不饱和的3-20元环状基团，优选3-10元环状基团，更优选3-6元环状基团；更加优选5-6元环状基团；所述杂环基含有1-19个碳原子，优选含有2-10个碳原子，更优选含有3-5个碳原子。杂环基的实例包括：氮杂环丙烷基、氧杂环丙烷基、氮杂环丁烷基、氧杂环丁烷基、1,4-苯并二噁烷基、1,3-苯并间二氧杂环戊烯基、二氢咪唑基、二氢吡喃基、二氢呋喃基、二噁烷基、亚乙基脲基、1,3-二氧杂环戊烷基、1,3-二噁烷基、1,4-二噁烷基、咪唑啉基、吲哚啉基、吗啉基、吡啶酮、2-吡咯烷酮、哌嗪基、高哌嗪基、哌啶基、高哌啶基、邻苯二甲酰亚胺基、丁二酰亚胺基、吡嗪基、吡唑啉基、吡咯啉基、四氢呋喃基、四氢吡喃基、四氢喹啉基、四氢噻吩基、噁烷基、氧硫杂环戊烷基、噻烷基等等。In the present disclosure, the term "heterocyclyl" refers to an optionally substituted group containing one or more (eg, 1, 2, 3 or 4) selected from N, O, S, SO, SO 2 or P Partially unsaturated or completely unsaturated 3-20-membered cyclic group of heteroatoms, preferably 3-10-membered cyclic group, more preferably 3-6-membered cyclic group; more preferably 5-6-membered cyclic group group; the heterocyclic group contains 1-19 carbon atoms, preferably 2-10 carbon atoms, and more preferably 3-5 carbon atoms. Examples of heterocyclyl include: aziridinyl, oxetanyl, azetidinyl, oxetanyl, 1,4-benzodioxanyl, 1,3-benzo Dioxolyl, dihydroimidazolyl, dihydropyranyl, dihydrofuryl, dioxanyl, ethyleneureido, 1,3-dioxolyl, 1, 3-dioxanyl, 1,4-dioxanyl, imidazolinyl, indolinyl, morpholinyl, pyridone, 2-pyrrolidone, piperazinyl, homopiperazinyl, piperidinyl, homo Piperidinyl, phthalimide, succinimide, pyrazinyl, pyrazolinyl, pyrrolinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydroquinolinyl, tetrahydrogen Thienyl, oxalkyl, oxathiolyl, thiyl, etc.
本公开中，术语“亚杂环基”是指杂环基进一步去掉一个氢原子形成的二价基团。In the present disclosure, the term "heterocyclylene" refers to a bivalent group formed by further removing one hydrogen atom from the heterocyclyl group.
本公开中，术语“杂环烷基”是指完全饱和的杂环基。In this disclosure, the term "heterocycloalkyl" refers to a fully saturated heterocyclyl group.
本公开中，术语“稠合芳基环烷基”是指上述芳基与环烷基稠和形成的基团。In the present disclosure, the term "fused arylcycloalkyl" refers to a group formed by the fusion of the above-mentioned aryl group and cycloalkyl group.
本公开中，术语“稠合芳基杂环基”是指上述芳基与杂环基稠和形成的基团。In the present disclosure, the term "fused arylheterocyclyl" refers to a group formed by the fusion of the above-mentioned aryl group and heterocyclyl.
本公开中，术语“稠合杂芳基环烷基”是指上述杂芳基与环烷基稠和形成的基团。In the present disclosure, the term "fused heteroarylcycloalkyl" refers to a group formed by the fusion of the above-mentioned heteroaryl and cycloalkyl.
本公开中，术语“稠合杂芳基杂环基”是指上述杂芳基与杂环基稠和形成的基团。In the present disclosure, the term "fused heteroarylheterocyclyl" refers to a group formed by the fusion of the above-mentioned heteroaryl and heterocyclyl.
本公开中，术语“芳基-烷基”是指上述芳基和烷基通过单键相连，并通过烷基连接其他部位。术语“杂芳基-烷基”、“芳基-环烷基”、“芳基-杂环基”、“环烷基-杂环基”、“杂环基-杂环基”、“芳基-芳基”、“芳基-杂芳基”、“杂芳基-芳基”、“杂芳基-杂芳基”以此类推。In the present disclosure, the term "aryl-alkyl" means that the above-mentioned aryl group and alkyl group are connected through a single bond, and other parts are connected through the alkyl group. The terms "heteroaryl-alkyl", "aryl-cycloalkyl", "aryl-heterocyclyl", "cycloalkyl-heterocyclyl", "heterocyclyl-heterocyclyl", "aryl-heterocyclyl" "aryl-aryl", "aryl-heteroaryl", "heteroaryl-aryl", "heteroaryl-heteroaryl" and so on.
本公开中，术语“杂环”是指可以任选地被取代的含有一个或多个(例如1、2、3或4个)选自N、O、S、SO、SO2或P的杂原子的部分不饱和或完全不饱和的3-20元环状基团，优选4-10元环状基团，3-7元环状基团，更优选3-6元环状基团，包括但不限于吡咯烷、二氢吡咯烷、四氢呋喃、哌啶、哌嗪、四氢吡喃、吡咯烷酮等环。 In the present disclosure, the term "heterocycle" refers to an optionally substituted heterocyclic ring containing one or more (eg, 1, 2, 3, or 4) selected from N, O, S, SO, SO 2, or P Atomically partially unsaturated or fully unsaturated 3-20-membered cyclic groups, preferably 4-10-membered cyclic groups, 3-7-membered cyclic groups, more preferably 3-6-membered cyclic groups, including But it is not limited to pyrrolidine, dihydropyrrolidine, tetrahydrofuran, piperidine, piperazine, tetrahydropyran, pyrrolidone and other rings.
本公开中，术语“碳环”是指环原子均为碳原子的部分不饱和或完全不饱和的3-10元环状基团，优选3-7元环状基团，更优选3-6元环状基团。任选地，环状结构中的碳原子可以被氧代，包括但不限于环丙烷、环丁烷等环。In the present disclosure, the term "carbocyclic ring" refers to a partially unsaturated or fully unsaturated 3-10-membered cyclic group in which all ring atoms are carbon atoms, preferably a 3-7-membered cyclic group, and more preferably a 3-6-membered cyclic group. cyclic group. Optionally, the carbon atoms in the cyclic structure can be oxo-substituted, including but not limited to cyclopropane, cyclobutane and other rings.
本公开中，术语“卤素”是指F、Cl、Br、I。In this disclosure, the term "halogen" refers to F, Cl, Br, I.
本公开中，术语“药物”指抑制或防止细胞的功能和/或引起细胞死亡或破坏的物质。In this disclosure, the term "drug" refers to a substance that inhibits or prevents the function of a cell and/or causes cell death or destruction.
本公开中，术语“连接体”是指将生物活性化合物片段(药物分子)与抗体部分连接起来的片段。在这点上，连接体在连接至抗体或其抗原结合片段之前(即连接体前体)具有可与抗体或其抗原结合片段的官能团形成键的官能团。In this disclosure, the term "linker" refers to a fragment that connects a biologically active compound fragment (drug molecule) to an antibody moiety. In this regard, the linker, prior to being attached to the antibody or antigen-binding fragment thereof (i.e., the linker precursor), has functional groups that can form bonds with functional groups of the antibody or antigen-binding fragment thereof.
本公开中，所述的“….生物活性化合物片段”是指本领域中均知的，抗体-药物偶联物(或称抗体偶联药物(antibody-drug conjugate,ADC))中，在肿瘤组织间或肿瘤细胞内连接子裂解/降解/酶切后，能够形成具有生物活性的药物(例如小分子细胞毒药物，所述药物包括其失去一个原子或原子团后的基团)或其衍生物(例如其前体)的部分(片段或基团)。为了避免歧义，“药物”并非仅指已获得医药监管部门审批的“药品”，还包括临床中，或者研发和学术研究中任何有潜在治疗生物活性的化合物。在本发明中，生物活性化合物包括本发明的Myc蛋白降解剂。In the present disclosure, the "... bioactive compound fragment" refers to the antibody-drug conjugate (or antibody-drug conjugate (ADC)) known in the art, which is used in tumors. After cleavage/degradation/enzyme cleavage of linkers between tissues or within tumor cells, biologically active drugs (such as small molecule cytotoxic drugs, which include groups after losing one atom or atomic group) or derivatives thereof ( For example, its precursor) part (fragment or group). To avoid ambiguity, "drugs" do not only refer to "drugs" that have been approved by the medical regulatory authorities, but also include any compounds with potential therapeutic biological activity in clinical practice, or in R&D and academic research. In the present invention, the biologically active compound includes the Myc protein degrading agent of the present invention.
本公开中，术语“接头单元”为抗体药物偶联物或药物连接体偶联物或连接子的组成部分，其作用是将和靶点结合的抗体或其抗原结合片段与抗体药物偶联物的其余部分相连。接头单元能够将Tb单元连接于L2上，具体实例包括但不限于(其中1位与抗体或其抗原结合片段相连、2位与L2或L3相连)： In the present disclosure, the term "linker unit" is a component of an antibody-drug conjugate or a drug-linker conjugate or a linker, and its function is to connect the antibody or antigen-binding fragment thereof that binds to the target with the antibody-drug conjugate. connected to the rest. The linker unit can connect the Tb unit to L 2. Specific examples include but are not limited to (where 1 position is connected to the antibody or its antigen-binding fragment, and 2 position is connected to L 2 or L 3 ):
本公开中，术语“连接单元”为抗体药物偶联物或药物连接体偶联物或连接子的组成部分，其作用是用来结合接头单元与氨基酸残基或由2-10个氨基酸残基组成的短肽。连接单元存在时能够将L1连接于L3。具体实例包括但不限于(其中1位与接头单元相连、2位与L3相连)： In the present disclosure, the term "linking unit" is a component of an antibody-drug conjugate or a drug-linker conjugate or a linker, which functions to combine the linker unit with an amino acid residue or consists of 2-10 amino acid residues. composed of short peptides. The presence of the connecting unit can connect L 1 to L 3 . Specific examples include but are not limited to (where 1 is connected to the connector unit and 2 is connected to L 3 ):
本公开中，术语“间隔单元”为抗体药物偶联物或药物连接体偶联物或连接子的组成部分，其作用是用来结合Myc蛋白降解剂片段与氨基酸残基或由2-10个氨基酸残基组成的短肽，或者其可提供附加的结构组成部分以进一步促进Myc蛋白降解剂从抗体药物偶联物的其余部分的释放。间隔单元存在时能够将D连接于L3。具体实例包括但不限于(其中1位与L3相连、2位与D相连)： In the present disclosure, the term "spacer unit" is a component of an antibody-drug conjugate or a drug-linker conjugate or a linker, which is used to bind the Myc protein degrader fragment to an amino acid residue or consists of 2-10 A short peptide consisting of amino acid residues, or which may provide additional structural components to further facilitate the release of the Myc protein-degrading agent from the remainder of the antibody drug conjugate. The presence of a spacer unit can connect D to L 3 . Specific examples include but are not limited to (where 1 bit is connected to L 3 and 2 bits are connected to D):
本公开中，术语“抗体”取其最广义的解释，包括完整的单克隆抗体、多克隆抗体以及由至少两个完整抗体形成的多特异性抗体(例如双特异性抗体)，只要它们具有所需的生物学活性。在本公开中，“抗体”和“免疫球蛋白”可以互换使用。In this disclosure, the term "antibody" is taken in its broadest interpretation and includes intact monoclonal antibodies, polyclonal antibodies, and multispecific antibodies (e.g., bispecific antibodies) formed from at least two intact antibodies, as long as they have the required required biological activity. In this disclosure, "antibody" and "immunoglobulin" are used interchangeably.
本文所用的术语“PSMA抗体”是指这样的抗体，所述抗体能够以足够的亲和力结合(人)PSMA以致所述抗体可以用作靶向(人)PSMA的治疗剂。在一个实施方案中，所述(人)PSMA抗体在体外或体内以高亲和力结合(人)PSMA。在一些实施方案中，所述结合例如通过放射性免疫测定(RIA)、生物膜薄层干涉测定法(BLI)、MSD测定法或表面等离子体共振法(SPR)或流式细胞术测量的。The term "PSMA antibody" as used herein refers to an antibody that is capable of binding (human) PSMA with sufficient affinity such that the antibody can be used as a therapeutic targeting (human) PSMA. In one embodiment, the (human) PSMA antibody binds (human) PSMA with high affinity in vitro or in vivo. In some embodiments, the binding is measured, for example, by radioimmunoassay (RIA), biofilm thin layer interferometry (BLI), MSD assay, or surface plasmon resonance (SPR), or flow cytometry.
术语“抗体片段”包括完整抗体的一部分。在优选的实施方案中，抗体片段为抗原结合片段。The term "antibody fragment" includes a portion of an intact antibody. In a preferred embodiment, the antibody fragment is an antigen-binding fragment.
“抗原结合片段”指与完整抗体不同的分子，其包含完整抗体的一部分且结合完整抗体所结合的抗原。抗体片段的例子包括但不限于Fv，Fab，Fab’，Fab’-SH，F(ab’)2；dAb(domain antibody)；线性抗体；单链抗体(例如scFv)；单结构域抗体例如VHH；双价抗体或其片段；或骆驼科抗体。"Antigen-binding fragment" refers to a molecule distinct from an intact antibody that contains a portion of an intact antibody and binds the antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2; dAb (domain antibody); linear antibodies; single chain antibodies (such as scFv); single domain antibodies such as VHH ; Diavalent antibodies or fragments thereof; or camelid antibodies.
在本公开中，术语“单克隆抗体”指抗体来自一群基本均一的抗体，即构成该集群的各抗体完全相同，除了可能存在的少量天然突变。单克隆抗体具有针对抗原的一个决定簇(表位)的高特异性，而与其相对的多克隆抗体则包含针对不同决定簇(表位)的不同抗体。除了特异性之外，单克隆抗体的优点还在于合成时可以不受其他抗体的污染。此处修饰语“单克隆”表示该抗体的特征在于来自一个基本均一的抗体群，而不应理解成需由特殊方法制得。In this disclosure, the term "monoclonal antibody" refers to an antibody derived from a population of antibodies that is substantially homogeneous, that is, the individual antibodies that make up the population are identical except for the possible presence of a small number of natural mutations. Monoclonal antibodies have high specificity for one determinant (epitope) of an antigen, whereas polyclonal antibodies contain different antibodies directed against different determinants (epitope). In addition to specificity, the advantage of monoclonal antibodies is that they can be synthesized without contamination from other antibodies. The modifier "monoclonal" here indicates that the antibody is characterized by being derived from a substantially homogeneous population of antibodies and should not be construed as requiring that it be prepared by a specific method.
在本公开的部分实施方案中，单克隆抗体还特别包括嵌合抗体，即重链和/或轻链的一部分与某种、某类或某亚类抗体相同或同源，其余部分则与另一种、 另一类或另一亚类抗体相同或同源，只要它们具有所需的生物学活性(参见例如US 4,816,567；和Morrison等人，1984，PNAS，81:6851-6855)。可用于本公开的嵌合抗体包括灵长类化(primatized)抗体，其包含来自非人灵长类(例如古猴、猩猩等)的可变区抗原结合序列和人恒定区序列。In some embodiments of the present disclosure, monoclonal antibodies also specifically include chimeric antibodies, that is, a part of the heavy chain and/or light chain is identical or homologous to a certain type, type or subtype of antibody, and the remaining part is identical to another type of antibody. A sort of, Antibodies of another class or subclass are the same or homologous as long as they have the desired biological activity (see, eg, US 4,816,567; and Morrison et al., 1984, PNAS, 81:6851-6855). Chimeric antibodies useful in the present disclosure include primatized antibodies comprising variable region antigen-binding sequences and human constant region sequences from non-human primates (eg, monkeys, orangutans, etc.).
在本公开中，非人(例如鼠)抗体的“人源化”形式指包含最少量非人免疫球蛋白序列的嵌合抗体。大多数人源化抗体是人接受者免疫球蛋白的超变区残基被置换成具有所需特异性、亲和力和功能的非人(例如小鼠、大鼠、兔或非人灵长类)超变区残基(供者抗体)。在一些实施方案中，人免疫球蛋白的框架区(FR)残基也被置换成非人残基。而且，人源化抗体还可以包含受者抗体或供者抗体中没有的残基。这些修饰是为了进一步优化抗体的性能。人源化抗体一般包含至少一个，通常是两个可变区，其中所有或几乎所有超变环(hypervanable loops)与非人免疫球蛋白的相对应，而FR则完全或几乎完全是人免疫球蛋白的序列。人源化抗体还可以包含免疫球蛋白恒定区(Fc，通常是人免疫球蛋白Fc)的至少一部分。有关细节参见例如Jones等人，1986，Nature，321:522-525；Riechmann等人，1988，Nature，332:323-329；和Presta，1992，Curr Op Struct Bwl 2:593-596。In this disclosure, a "humanized" form of a non-human (eg, murine) antibody refers to a chimeric antibody that contains a minimal amount of non-human immunoglobulin sequence. Most humanized antibodies are human recipient immunoglobulins whose hypervariable region residues have been replaced with non-human ones (e.g., mouse, rat, rabbit, or non-human primate) that have the desired specificity, affinity, and function. Hypervariable region residues (donor antibody). In some embodiments, framework region (FR) residues of human immunoglobulins are also replaced with non-human residues. Furthermore, humanized antibodies may also contain residues that are not found in either the recipient antibody or the donor antibody. These modifications are intended to further optimize the antibody's performance. Humanized antibodies generally contain at least one, and usually two, variable regions, in which all or nearly all hypervanable loops correspond to those of non-human immunoglobulins, while the FRs are entirely or almost entirely human immunoglobulins. Protein sequence. The humanized antibody may also comprise at least a portion of an immunoglobulin constant region (Fc, typically a human immunoglobulin Fc). For details see, for example, Jones et al., 1986, Nature, 321:522-525; Riechmann et al., 1988, Nature, 332:323-329; and Presta, 1992, Curr Op Struct Bwl 2:593-596.
完整抗体可根据重链恒定区的氨基酸序列分为不同的“类”。主要的五类是IgA、IgD、IgE、IgG和IgM，其中几类还可以分为不同的“亚类”(同种型)，例如IgG1、IgG2、IgG3、IgG4、IgA1和IgA2。抗体不同类的重链恒定区分别称为α、β、ε、γ和μ。免疫球蛋白不同类的亚基结构和三维构型是本领域中公知的。Intact antibodies can be divided into different "classes" based on the amino acid sequence of the heavy chain constant region. The five main classes are IgA, IgD, IgE, IgG and IgM, several of which can also be divided into different "subclasses" (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. The heavy chain constant regions of different classes of antibodies are called α, β, ε, γ, and μ. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known in the art.
本公开所用的单克隆抗体可以由许多方法生产。例如，用于本公开的单克隆抗体可以通过杂交瘤方法，使用许多物种(包括小鼠、仓鼠、大鼠和人的细胞)获得(参见例如Kohler等人，1975，Nature，256:495)，或者通过重组DNA技术制得(参见例如US 4,816,567)，或者从噬菌体抗体库中分离得到(参见例如Clackson等人，1991，Nature，352:624-628；和Marks等人，1991，Journal of Molecular Biology，222:581-597)。Monoclonal antibodies used in the present disclosure can be produced by a number of methods. For example, monoclonal antibodies useful in the present disclosure can be obtained by hybridoma methods using cells from a number of species, including mouse, hamster, rat, and human (see, e.g., Kohler et al., 1975, Nature, 256:495). Either prepared by recombinant DNA technology (see, e.g., US 4,816,567), or isolated from a phage antibody library (see, e.g., Clackson et al., 1991, Nature, 352:624-628; and Marks et al., 1991, Journal of Molecular Biology , 222:581-597).
术语“药物与抗体比率”或“DAR”是指偶联于本文所述的抗体部分(Tb)的小分子药物部分(D)与抗体部分的数量比例。ADC的DAR可以在1到20的范围内，但是取决于抗体上的连接位点的数量，更高的负载也是可能的。提及负载到单个抗体上的药物的数量时，或可替代地，提及一组ADC的平均或均值DAR时，可以使用术语DAR。DAR也可被计算为产品中分子群体的平均DAR，即通过检 测方法(例如通过常规方法如质谱法、ELISA测定、电泳和/或HPLC)测得的产品中偶联于本文所述的Ab部分的小分子药物部分(D)与Ab部分的总体比例(摩尔比例)，此DAR在文中称为平均DAR。在一些实施方案中，本发明的抗体药物偶联物的平均DAR值是1.0-20.0，例如1.0-18.0、1.0-16.0、2.0-14.0、3.0-12.0、4.0-10.0、5.0-9.0、6.0-8.0、1.0-8.0，2.0-6.0，例如1.0、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、3、3.1、3.2、3.3、3.4、3.5、3.6、3.7、3.8、3.9、4、4.1、4.2、4.3、4.4、4.5、4.6、4.7、4.8、4.9、5.0、5.1、5.2、5.3、5.4、5.5、5.6、5.7、5.8、5.9、6.0、6.1、6.2、6.3、6.4、6.5、6.6、6.7、6.8、6.9、7.0、7.1、7.2、7.3、7.4、7.5、7.6、7.7、7.8.0、7.9、8、8.1、8.2、8.3、8.4、8.5、8.6、8.7、8.8、8.9、9.0、9.1、9.2、9.3、9.4、9.5、9.6、9.7、9.8、9.9、10.0、12.0和16.0，以这些数值中的两个作为端点的范围。The term "drug to antibody ratio" or "DAR" refers to the quantitative ratio of small molecule drug moiety (D) to antibody moiety coupled to the antibody moiety (Tb) described herein. The DAR of an ADC can range from 1 to 20, but depending on the number of attachment sites on the antibody, higher loadings are also possible. The term DAR may be used when referring to the amount of drug loaded onto a single antibody, or alternatively, when referring to the average or mean DAR of a group of ADCs. DAR can also be calculated as the average DAR of the population of molecules in the product, i.e. The overall ratio (molar) of the small molecule drug moiety (D) coupled to the Ab moiety described herein to the Ab moiety in the product as measured by conventional methods such as mass spectrometry, ELISA assays, electrophoresis and/or HPLC. ratio), this DAR is called the average DAR in the text. In some embodiments, the average DAR value of the antibody drug conjugate of the invention is 1.0-20.0, such as 1.0-18.0, 1.0-16.0, 2.0-14.0, 3.0-12.0, 4.0-10.0, 5.0-9.0, 6.0- 8.0, 1.0-8.0, 2.0-6.0, such as 1.0, 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 ,3,3.1,3.2,3.3,3.4,3.5,3.6,3.7,3.8,3.9,4,4.1,4.2,4.3,4.4,4.5,4.6,4.7,4.8,4.9,5.0,5.1,5.2,5.3,5.4 ,5.5,5.6,5.7,5.8,5.9,6.0,6.1,6.2,6.3,6.4,6.5,6.6,6.7,6.8,6.9,7.0,7.1,7.2,7.3,7.4,7.5,7.6,7.7,7.8.0 ,7.9,8,8.1,8.2,8.3,8.4,8.5,8.6,8.7,8.8,8.9,9.0,9.1,9.2,9.3,9.4,9.5,9.6,9.7,9.8,9.9,10.0,12.0 and 16.0, to Two of these values serve as the endpoints of the range.
术语“约”在与数字数值联合使用时意为涵盖具有比指定数字数值小10％的下限和比指定数字数值大10％的上限的范围内的数字数值。The term "about" when used in conjunction with a numerical value is meant to encompass a range of numerical values having a lower limit that is 10% less than the specified numerical value and an upper limit that is 10% greater than the specified numerical value.
如本文所用，术语“和/或”意指可选项中的任一项或可选项的两项或多项。As used herein, the term "and/or" means any one of the options or two or more of the options.
如本文所用，术语“包含”或“包括”意指包括所述的要素、整数或步骤，但是不排除任意其他要素、整数或步骤。在本文中，当使用术语“包含”或“包括”时，除非另有指明，否则也涵盖由所述及的要素、整数或步骤组合的情形。例如，当提及“包含”某个具体序列的抗体可变区时，也旨在涵盖由该具体序列组成的抗体可变区。As used herein, the term "comprises" or "includes" means the inclusion of the stated element, integer or step, but not the exclusion of any other element, integer or step. When the term "comprises" or "includes" is used herein, combinations of the stated elements, integers, or steps are also encompassed unless otherwise specified. For example, when reference is made to an antibody variable region that "comprises" a particular sequence, it is also intended to encompass antibody variable regions that consist of that particular sequence.
本公开所用试剂和原料均市售可得。The reagents and raw materials used in this disclosure are all commercially available.
发明的有益效果Beneficial effects of the invention
本公开所述蛋白降解剂类生物活性化合物的抗体偶联药物能够用于多种血液瘤和实体瘤的治疗，并且具有比对应的蛋白降解剂类生物活性化合物更好的治疗效果和/或更低的副作用；The antibody-conjugated drugs of the protein-degrading bioactive compounds disclosed in the present disclosure can be used for the treatment of various hematological tumors and solid tumors, and have better therapeutic effects and/or better therapeutic effects than the corresponding protein-degrading bioactive compounds. Low side effects;
本发明的偶联物具有更好的溶解度和优异的化学稳定性，具有高的药物-抗体比； The conjugate of the present invention has better solubility and excellent chemical stability, and has a high drug-antibody ratio;
本发明的偶联物中的连接子具有高血浆稳定性，但同时又能在肿瘤微环境中(肿瘤细包内和肿瘤细胞外均可)裂解，因此可以在抗原低表达或者抗原不表达肿瘤中产生好的抗肿瘤效果；The linker in the conjugate of the present invention has high plasma stability, but at the same time can be cleaved in the tumor microenvironment (both within tumor cells and outside tumor cells), so it can be used in tumors with low antigen expression or no antigen expression. produce good anti-tumor effects;
本发明的偶联物(包括ADC)具有更好的肿瘤组织靶向性，即在肿瘤微环境中的富集能力，增加了生物活性分子在瘤内和血液浓度比，降低了偶联物的机理相关的毒性，具有更高的治疗指数；The conjugate (including ADC) of the present invention has better tumor tissue targeting, that is, the ability to enrich in the tumor microenvironment, increases the concentration ratio of bioactive molecules in the tumor and blood, and reduces the concentration ratio of the conjugate. Mechanism-related toxicities with higher therapeutic index;
本发明的偶联物具有很高的在体内循环中的稳定性，减少了药物分子在非靶组织中的脱落，减少了非靶组织中毒素脱落引起的“off-target”毒性；The conjugate of the present invention has high stability in body circulation, reduces the shedding of drug molecules in non-target tissues, and reduces the "off-target" toxicity caused by the shedding of toxins in non-target tissues;
所述偶联物的生物活性分子具有更高的抗肿瘤细胞活性，因此具有优异的旁观者效应(by-stander effect)，所述偶联物能够更有效地杀死抗原高表达肿瘤细胞以及肿瘤组织中抗原低表达或抗原不表达的肿瘤细胞；和/或The bioactive molecules of the conjugate have higher anti-tumor cell activity and therefore have excellent by-stander effect. The conjugate can more effectively kill tumor cells with high antigen expression and tumors. Tumor cells with low or no antigen expression in tissues; and/or
本公开的药物-连接子偶联物，利用其连接子在肿瘤微环境中的胞外裂解能力，可以和没有细胞内吞能力的抗体组成抗体偶联药物，这类抗体偶联药物依然具有高抗肿瘤活性。The disclosed drug-linker conjugate utilizes the extracellular cleavage ability of its linker in the tumor microenvironment to form an antibody-conjugated drug with an antibody without cell endocytosis ability. This type of antibody-conjugated drug still has high Antitumor activity.
附图说明Description of the drawings
图1：显示化合物D-2和化合物D-4对HL60细胞中的c-Myc和GSPT1蛋白的降解能力。Figure 1: Shows the degradation ability of compound D-2 and compound D-4 on c-Myc and GSPT1 proteins in HL60 cells.
图2：显示本发明ADC能显著降解22RV1细胞中的c-Myc蛋白。Figure 2: shows that the ADC of the present invention can significantly degrade c-Myc protein in 22RV1 cells.
具体实施方式Detailed ways
以下通过具体实施方式的描述对本公开作进一步说明，但这并非是对本公开的限制。本领域技术人员根据本公开的教导，可以做出各种修改或改进，而不脱离本公开的基本思想和范围。所用试剂或仪器未注明生产厂商者，均为可以通过市购获得的常规产品。The present disclosure will be further described below through the description of specific embodiments, but this is not intended to limit the present disclosure. Those skilled in the art can make various modifications or improvements based on the teachings of the present disclosure without departing from the basic idea and scope of the present disclosure. If the manufacturer of the reagents or instruments used is not indicated, they are all conventional products that can be purchased commercially.
本公开中的缩写具有以下含义：

Abbreviations in this disclosure have the following meanings:

制备方案Preparation protocol
以下的实施例中记载的化合物的结构通过核磁共振(1H NMR)或质谱(MS)来确定。The structures of the compounds described in the following examples were determined by nuclear magnetic resonance ( 1 H NMR) or mass spectrometry (MS).
核磁共振(1H NMR)的测定仪器使用Bruker 400MHz核磁共振仪；测定溶剂为氘代甲醇(CD3OD)、氘代氯仿(CDCl3)或六氘代二甲基亚砜(DMSO-d6)；内标物质为四甲基硅烷(TMS)。The measuring instrument of nuclear magnetic resonance ( 1 H NMR) uses a Bruker 400MHz nuclear magnetic resonance instrument; the measuring solvent is deuterated methanol (CD 3 OD), deuterated chloroform (CDCl 3 ) or hexadeuterated dimethyl sulfoxide (DMSO-d 6 ); the internal standard substance is tetramethylsilane (TMS).
实施例中使用的核磁共振(NMR)图谱中的缩写示于以下。The abbreviations in the nuclear magnetic resonance (NMR) spectra used in the examples are shown below.
s：单峰(singlet)、d：二重峰(doublet)、t：三重峰(triplet)、q：四重峰(quartet)、dd：双二重峰(double doublet)、qd：四二重峰(quartet doublet)、ddd：双双二重峰(double double doublet)、ddt：双双三重峰(double double triplet)、dddd：双双双二重峰(double double double doublet)、m：多重峰(multiplet)、br：宽峰(broad)、J：偶合常数、Hz：赫兹、DMSO-d6：氘化二甲基亚砜。δ值用ppm值表示。s: singlet, d: doublet, t: triplet, q: quartet, dd: double doublet, qd: quadruplet Peak (quartet doublet), ddd: double double doublet (double double doublet), ddt: double double triplet (double double triplet), dddd: double double doublet (double double double doublet), m: multiplet (multiplet) , br: broad, J: coupling constant, Hz: Hertz, DMSO-d 6 : deuterated dimethyl sulfoxide. The delta value is expressed as ppm value.
质谱(MS)的测定仪器使用Agilent(ESI)质谱仪，型号为Agilent 6120B。The mass spectrometry (MS) measurement instrument used was an Agilent (ESI) mass spectrometer, model Agilent 6120B.
实施例1.小分子蛋白降解剂的合成Example 1. Synthesis of small molecule protein degradation agent
实施例1.1：2-(2,6-二氧代哌啶-3-基)-N-(4-(((2-羟基乙基)(苯乙基)氨基)甲基)苄基)-1-氧代异吲哚啉-5-甲酰胺(D-1)的合成
Example 1.1: 2-(2,6-dioxopiperidin-3-yl)-N-(4-(((2-hydroxyethyl)(phenylethyl)amino)methyl)benzyl)- Synthesis of 1-oxoisoindoline-5-carboxamide (D-1)
步骤一：D-1-1的合成Step 1: Synthesis of D-1-1
将4-溴-2-甲基苯甲酸甲酯(30.0g,131mmol)溶于300mL四氯化碳中，向其中加入NBS(28g,131mmol)和AIBN(6.4g,40mmol)，80℃反应1h。加入150mL MTBE稀释后，过滤。滤液用饱和碳酸氢钠水溶液洗(30mL*3)。有机相干燥旋干，得化合物D-1-1(48g,粗品)。Dissolve 4-bromo-2-methylbenzoic acid methyl ester (30.0g, 131mmol) in 300mL carbon tetrachloride, add NBS (28g, 131mmol) and AIBN (6.4g, 40mmol), and react at 80°C for 1 hour. . Add 150mL MTBE to dilute and filter. Wash the filtrate with saturated sodium bicarbonate aqueous solution (30mL*3). The organic phase was dried and spin-dried to obtain compound D-1-1 (48g, crude product).
步骤二：D-1-2的合成Step 2: Synthesis of D-1-2
将化合物D-1-1(48g,131mmol)溶于450mL DMF中，向其中加入3-氨基哌啶-2,6-二酮盐酸盐(28g,172mmol)和碳酸钾(65g,469mmol)，70℃反应1h。反应液过滤，滤液旋去大部分溶剂。用200mL二氯甲烷/水(1:1)打浆后，抽滤。滤饼干燥后得化合物D-1-2(19g)。Compound D-1-1 (48g, 131mmol) was dissolved in 450mL DMF, and 3-aminopiperidine-2,6-dione hydrochloride (28g, 172mmol) and potassium carbonate (65g, 469mmol) were added thereto. React at 70°C for 1 hour. The reaction solution was filtered, and most of the solvent was removed from the filtrate. After beating with 200mL methylene chloride/water (1:1), suction filter. After drying the filter cake, compound D-1-2 (19g) was obtained.
步骤三：D-1-3的合成Step 3: Synthesis of D-1-3
将化合物D-1-2(10g,31mmol)溶于80mL DMF中，依次向其中加入甲酸(5g,109mmol)，醋酸钯(209mg,0.93mmol)，DCC(1.3g,6.2mmol)，三乙胺(6.3g,62mmol)和Xantphos(539mg,0.93mmol)。氮气保护下，100℃反应3h。反应液过滤后，滤液减压蒸除大部分溶剂。用50mL二氯甲烷打浆后，抽滤。滤饼干燥后得化合物D-1-3(7.8g)。 Dissolve compound D-1-2 (10g, 31mmol) in 80mL DMF, and add formic acid (5g, 109mmol), palladium acetate (209mg, 0.93mmol), DCC (1.3g, 6.2mmol), and triethylamine in sequence. (6.3g, 62mmol) and Xantphos (539mg, 0.93mmol). Under nitrogen protection, react at 100°C for 3 hours. After the reaction solution was filtered, most of the solvent was evaporated from the filtrate under reduced pressure. After beating with 50 mL of methylene chloride, filter with suction. After drying the filter cake, compound D-1-3 (7.8g) was obtained.
1H NMR(400MHz,DMSO-d6)δ11.02(s,1H),8.17(s,1H),8.00(d,J＝7.9Hz,1H),7.96(s,1H),7.84(d,J＝7.9Hz,1H),5.14(dd,J＝13.3,5.1Hz,1H),4.55-4.37(m,2H),2.99-2.84(m,1H),2.69-2.58(m,1H),2.46-2.38(m,1H),2.07-1.95(m,1H). 1 H NMR (400MHz, DMSO-d 6 ) δ11.02 (s, 1H), 8.17 (s, 1H), 8.00 (d, J = 7.9Hz, 1H), 7.96 (s, 1H), 7.84 (d, J＝7.9Hz,1H),5.14(dd,J＝13.3,5.1Hz,1H),4.55-4.37(m,2H),2.99-2.84(m,1H),2.69-2.58(m,1H),2.46 -2.38(m,1H),2.07-1.95(m,1H).
步骤四：D-1-4的合成Step 4: Synthesis of D-1-4
将化合物D-1-3(730mg,2.53mmol),1-(N-Boc-氨基甲基)-4-(氨基甲基)苯(600mg,2.53mmol),DIPEA(1.96g,15.18mmol)依次加入到20mL DMF中。冰浴下向其中加入HATU(1.15g,3.04mmol)。常温反应1h。向其中加入120mL水/乙酸乙酯(1:1),打浆1h后过滤。滤饼用乙酸乙酯洗后干燥得化合物D-1-4(710mg)。Compound D-1-3 (730mg, 2.53mmol), 1-(N-Boc-aminomethyl)-4-(aminomethyl)benzene (600mg, 2.53mmol), DIPEA (1.96g, 15.18mmol) were added in sequence Add to 20mL DMF. HATU (1.15g, 3.04mmol) was added thereto under ice bath. Reaction at room temperature for 1 hour. Add 120 mL water/ethyl acetate (1:1) to it, beat for 1 hour and then filter. The filter cake was washed with ethyl acetate and dried to obtain compound D-1-4 (710 mg).
1H NMR(400MHz,DMSO-d6)δ11.01(s,1H),9.21(t,J＝5.8Hz,1H),8.09(s,1H),8.01(d,J＝8.3Hz,1H),7.82(d,J＝8.0Hz,1H),7.36(s,1H),7.28(d,J＝7.8Hz,2H),7.19(d,J＝7.9Hz,2H),5.14(dd,J＝13.3,5.0Hz,1H),4.56-4.36(m,4H),4.09(d,J＝5.9Hz,2H),2.97-2.85(m,1H),2.69-2.57(m,1H),2.46-2.37(m,1H),2.08-1.98(m,1H),1.38(s,9H). 1 H NMR (400MHz, DMSO-d 6 ) δ11.01 (s, 1H), 9.21 (t, J = 5.8Hz, 1H), 8.09 (s, 1H), 8.01 (d, J = 8.3Hz, 1H) ,7.82(d,J＝8.0Hz,1H),7.36(s,1H),7.28(d,J＝7.8Hz,2H),7.19(d,J＝7.9Hz,2H),5.14(dd,J＝ 13.3,5.0Hz,1H),4.56-4.36(m,4H),4.09(d,J＝5.9Hz,2H),2.97-2.85(m,1H),2.69-2.57(m,1H),2.46-2.37 (m,1H),2.08-1.98(m,1H),1.38(s,9H).
步骤五：D-1-5的合成Step 5: Synthesis of D-1-5
化合物D-1-4(710mg,1.4mmol)溶于二氯甲烷(20mL)中，加入HCl-Dioxane(10mL，4M)，室温反应1h。旋干溶剂后，得化合物D-1-5盐酸盐(700mg)，直接用于下一步反应。Compound D-1-4 (710 mg, 1.4 mmol) was dissolved in dichloromethane (20 mL), HCl-Dioxane (10 mL, 4 M) was added, and the reaction was carried out at room temperature for 1 h. After spinning the solvent dry, compound D-1-5 hydrochloride (700 mg) was obtained, which was directly used in the next reaction.
步骤六：D-1-6的合成Step 6: Synthesis of D-1-6
将化合物D-1-5(700mg，1.4mmol)，苯乙醛(185mg，1.5mmol)溶于甲醇(10mL)中。室温搅拌5min后，加入NaBH3CN(132mg，2.1mmol)，室温继续反应2h。反应液加NaHCO3水溶液(30mL)后，用二氯甲烷萃取(35mL*3)。有机相合并干燥后，旋干溶剂。粗品经柱层析纯化(洗脱剂为二氯甲烷：甲醇＝15:1)，得化合物D-1-6(600mg)。Compound D-1-5 (700 mg, 1.4 mmol) and phenylacetaldehyde (185 mg, 1.5 mmol) were dissolved in methanol (10 mL). After stirring at room temperature for 5 min, NaBH 3 CN (132 mg, 2.1 mmol) was added, and the reaction was continued at room temperature for 2 h. After adding NaHCO 3 aqueous solution (30mL) to the reaction solution, extract with dichloromethane (35mL*3). After the organic phases were combined and dried, the solvent was spun off. The crude product was purified by column chromatography (eluent: dichloromethane: methanol = 15:1) to obtain compound D-1-6 (600 mg).
ESI-MS[M+H]+:511.2。ESI-MS[M+H] + :511.2.
步骤七：D-1的合成Step 7: Synthesis of D-1
化合物D-1-6(600mg，1.17mmol)，2-溴乙醇(293mg，3.51mmol)，以及DIPEA(452mg，3.5mmol)溶于DMF(15mL)中，80℃搅拌过夜。加水(50mL)淬灭反应，水相用乙酸乙酯萃取(50mL*3)后，有机相合并再用饱和食盐水洗(30mL*3)。有机相用无水硫酸钠干燥，过滤，旋干溶剂。粗品用柱层析纯化(洗脱剂为二氯甲烷：甲醇＝15:1)，得化合物D-1(200mg)。 Compound D-1-6 (600 mg, 1.17 mmol), 2-bromoethanol (293 mg, 3.51 mmol), and DIPEA (452 mg, 3.5 mmol) were dissolved in DMF (15 mL), and stirred at 80°C overnight. Add water (50 mL) to quench the reaction, extract the aqueous phase with ethyl acetate (50 mL*3), combine the organic phases and wash with saturated brine (30 mL*3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the solvent was spun dry. The crude product was purified by column chromatography (eluent: dichloromethane: methanol = 15:1) to obtain compound D-1 (200 mg).
ESI-MS[M+H]+:555.3.ESI-MS[M+H] + :555.3.
1H NMR(400MHz,DMSO-d6)δ11.01(s,1H),9.22(s,1H),8.10(s,1H),8.01(d,J＝8.0Hz,1H),7.82(d,J＝7.9Hz,1H),7.39-7.10(m,9H),5.13(dd,J＝13.2,5.1Hz,1H),4.60-4.32(m,4H),3.78-3.48(m,6H),2.98-2.86(m,1H),2.83-2.53(m,5H),2.46-2.31(m,1H),2.09-1.96(m,1H). 1 H NMR (400MHz, DMSO-d 6 ) δ11.01 (s, 1H), 9.22 (s, 1H), 8.10 (s, 1H), 8.01 (d, J = 8.0Hz, 1H), 7.82 (d, J＝7.9Hz,1H),7.39-7.10(m,9H),5.13(dd,J＝13.2,5.1Hz,1H),4.60-4.32(m,4H),3.78-3.48(m,6H),2.98 -2.86(m,1H),2.83-2.53(m,5H),2.46-2.31(m,1H),2.09-1.96(m,1H).
实施例1.2：N-(4-(((3,4-二氯苯基)(2-羟基乙基)氨基)甲基)苄基)-2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-甲酰胺(D-2)的合成
Example 1.2: N-(4-(((3,4-dichlorophenyl)(2-hydroxyethyl)amino)methyl)benzyl)-2-(2,6-dioxopiperidine- Synthesis of 3-yl)-1-oxoisoindoline-5-carboxamide (D-2)
除了在步骤六中用3,4-二氯苯乙醛代替苯乙醛外，参考D-1的合成方法，得到D-2。Except using 3,4-dichlorophenylacetaldehyde instead of phenylacetaldehyde in step 6, refer to the synthesis method of D-1 to obtain D-2.
ESI-MS[M+H]+:623.2.ESI-MS[M+H] + :623.2.
1H NMR(400MHz,DMSO-d6)δ11.01(s,1H),9.20(t,J＝6.1Hz,1H),8.10(s,1H),8.01(d,J＝7.7Hz,1H),7.82(d,J＝7.9Hz,1H),7.49-7.46(m,2H),7.25-7.14(m,5H),5.14(dd,J＝13.3,5.1Hz,1H),4.55-4.28(m,5H),3.62(s,2H),3.46-3.39(m,2H),2.98-2.85(m,1H),2.77-2.65(m,4H),2.63-2.51(m,3H),2.42(dd,J＝12.9,4.5Hz,1H),2.06-1.98(m,1H). 1 H NMR (400MHz, DMSO-d 6 ) δ11.01(s,1H),9.20(t,J＝6.1Hz,1H),8.10(s,1H),8.01(d,J＝7.7Hz,1H) ,7.82(d,J＝7.9Hz,1H),7.49-7.46(m,2H),7.25-7.14(m,5H),5.14(dd,J＝13.3,5.1Hz,1H),4.55-4.28(m ,5H),3.62(s,2H),3.46-3.39(m,2H),2.98-2.85(m,1H),2.77-2.65(m,4H),2.63-2.51(m,3H),2.42(dd ,J＝12.9,4.5Hz,1H),2.06-1.98(m,1H).
实施例1.3：2-(2,6-二氧代哌啶-3-基)-N-(4-(((2-羟基乙基)(2-(萘-1-基)乙基)氨基)甲基)苄基)-1-氧代异吲哚啉-5-甲酰胺(D-3)的合成
Example 1.3: 2-(2,6-dioxopiperidin-3-yl)-N-(4-(((2-hydroxyethyl)(2-(naphth-1-yl)ethyl)amino) Synthesis of )methyl)benzyl)-1-oxoisoindoline-5-carboxamide (D-3)
除了在步骤六中用2-(萘-1-基)乙醛代替苯乙醛外，参考D-1的合成方法，得到D-3。Except using 2-(naphth-1-yl)acetaldehyde instead of phenylacetaldehyde in step 6, refer to the synthesis method of D-1 to obtain D-3.
ESI-MS[M+H]+:605.3.ESI-MS[M+H] + :605.3.
1H NMR(400MHz,DMSO-d6)δ11.01(s,1H),9.22(t,J＝6.0Hz,1H),8.11(s,1H),8.03(d,J＝8.1Hz,1H),7.89-7.80(m,3H),7.73(d,J＝7.9Hz,1H),7.45-7.36(m,3H),7.35-7.24(m,5H),5.14(dd,J＝13.3,5.1Hz,1H),4.58-4.34(m,5H),3.74 (s,2H),3.50(t,J＝6.4Hz,2H),3.23-3.14(m,2H),2.98-2.83(m,1H),2.81-2.72(m,2H),2.71-2.56(m,3H),2.46-2.32(m,1H),2.06-2.00(m,1H). 1 H NMR (400MHz, DMSO-d 6 ) δ11.01 (s, 1H), 9.22 (t, J = 6.0Hz, 1H), 8.11 (s, 1H), 8.03 (d, J = 8.1Hz, 1H) ,7.89-7.80(m,3H),7.73(d,J＝7.9Hz,1H),7.45-7.36(m,3H),7.35-7.24(m,5H),5.14(dd,J＝13.3,5.1Hz ,1H),4.58-4.34(m,5H),3.74 (s,2H),3.50(t,J＝6.4Hz,2H),3.23-3.14(m,2H),2.98-2.83(m,1H),2.81-2.72(m,2H),2.71-2.56(m ,3H),2.46-2.32(m,1H),2.06-2.00(m,1H).
实施例1.4：2-(2,6-二氧代哌啶-3-基)-N-(4-((2-羟基-N-(2-(萘-1-基)乙基)乙酰胺)甲基)苄基)-1-氧代异吲哚啉-5-甲酰胺(D-4)的合成
Example 1.4: 2-(2,6-dioxopiperidin-3-yl)-N-(4-((2-hydroxy-N-(2-(naphthalen-1-yl)ethyl)ethyl)acetamide Synthesis of )methyl)benzyl)-1-oxoisoindoline-5-carboxamide (D-4)
步骤一：D-4-1的合成Step 1: Synthesis of D-4-1
将2-(1-萘基)乙醇(860mg,5mmol)溶于18mL DCM中，于0℃下向其中分批加入Dess-Martin试剂(3.18g,7.5mmol)，室温下反应2h，直接过滤。滤液加NaHCO3水溶液(30mL)后，用二氯甲烷萃取(30mL*3)。有机相合并后，用无水硫酸钠干燥，过滤，旋干溶剂。粗品用柱层析纯化(洗脱剂为石油醚：乙酸乙酯＝100:1–15:1)，得到化合物D-4-1(690mg)。Dissolve 2-(1-naphthyl)ethanol (860 mg, 5 mmol) in 18 mL DCM, add Dess-Martin reagent (3.18 g, 7.5 mmol) in batches at 0°C, react at room temperature for 2 h, and filter directly. After adding NaHCO 3 aqueous solution (30mL) to the filtrate, extract with dichloromethane (30mL*3). After the organic phases were combined, they were dried over anhydrous sodium sulfate, filtered, and the solvent was spin-dried. The crude product was purified by column chromatography (eluent was petroleum ether: ethyl acetate = 100:1–15:1) to obtain compound D-4-1 (690 mg).
步骤二：D-4-2的合成Step 2: Synthesis of D-4-2
将化合物D-4-1(690mg,4.1mmol)和1-(N-Boc-氨基甲基)-4-(氨基甲基)苯(968mg,4.1mmol)溶于12mL甲醇中，加入NaBH3CN(504mg,8mmol)和AcOH(100mg)，室温反应4h。加NaHCO3水溶液(30mL)后，用二氯甲烷萃取(35mL*3)。有机相合并后，用无水硫酸钠干燥，过滤，旋干溶剂。粗品用柱层析纯化(洗脱剂为二氯甲烷：甲醇＝20:1)，得化合物D-4-2(540mg)。Compound D-4-1 (690 mg, 4.1 mmol) and 1-(N-Boc-aminomethyl)-4-(aminomethyl)benzene (968 mg, 4.1 mmol) were dissolved in 12 mL of methanol, and NaBH 3 CN was added (504mg, 8mmol) and AcOH (100mg), react at room temperature for 4h. After adding NaHCO 3 aqueous solution (30mL), extract with dichloromethane (35mL*3). After the organic phases were combined, they were dried over anhydrous sodium sulfate, filtered, and the solvent was spin-dried. The crude product was purified by column chromatography (eluent: dichloromethane: methanol = 20:1) to obtain compound D-4-2 (540 mg).
ESI-MS[M+H]+:391.2。ESI-MS[M+H] + :391.2.
步骤三：D-4-3的合成Step 3: Synthesis of D-4-3
将化合物D-4-2(540mg,1.38mmol)和羟基乙酸(106mg,1.4mmol)溶于10mL二氯甲烷中，向其中加入HATU(630mg,1.66mmol)和TEA(420mg,4.2 mmol)，室温反应2h。加水(30mL)淬灭反应，水相用二氯甲烷萃取(35mL*3)。有机相合并后，用无水硫酸钠干燥，过滤，旋干溶剂。粗品用柱层析纯化(洗脱剂为二氯甲烷：甲醇＝20:1)，得化合物D-4-3(406mg)。Compound D-4-2 (540 mg, 1.38 mmol) and glycolic acid (106 mg, 1.4 mmol) were dissolved in 10 mL of methylene chloride, and HATU (630 mg, 1.66 mmol) and TEA (420 mg, 4.2 mmol), react at room temperature for 2 h. Add water (30 mL) to quench the reaction, and extract the aqueous phase with dichloromethane (35 mL*3). After the organic phases were combined, they were dried over anhydrous sodium sulfate, filtered, and the solvent was spin-dried. The crude product was purified by column chromatography (eluent: dichloromethane: methanol = 20:1) to obtain compound D-4-3 (406 mg).
ESI-MS[M+H]+:449.2。ESI-MS[M+H] + :449.2.
步骤四：D-4-4的合成Step 4: Synthesis of D-4-4
将化合物D-4-3(406mg,0.9mmol)溶于4mL四氢呋喃中，向其中加入HCl-Dioxane(3mL，4M)，室温反应4h。旋干溶剂，得化合物D-4-4盐酸盐(400mg,粗品)直接用于下一步反应。Compound D-4-3 (406 mg, 0.9 mmol) was dissolved in 4 mL of tetrahydrofuran, HCl-Dioxane (3 mL, 4 M) was added thereto, and the reaction was carried out at room temperature for 4 h. The solvent was spun dry to obtain compound D-4-4 hydrochloride (400 mg, crude product), which was directly used in the next reaction.
步骤五：D-4的合成Step 5: Synthesis of D-4
将化合物D-4-4(400mg,0.89mmol)和化合物D-1-3(259mg,0.9mmol)溶于10mL二氯甲烷中，向其中加入HATU(410mg,1.1mmol)和三乙胺(270mg,2.7mmol)，室温反应1.5h。加水(50mL)淬灭反应，水相用二氯甲烷萃取(35mL*3)。有机相合并后，用无水硫酸钠干燥，过滤，旋干溶剂。粗品用柱层析纯化(洗脱剂为二氯甲烷：甲醇＝15:1)后，得到白色固体化合物D-4(127mg)。Compound D-4-4 (400 mg, 0.89 mmol) and compound D-1-3 (259 mg, 0.9 mmol) were dissolved in 10 mL of methylene chloride, and HATU (410 mg, 1.1 mmol) and triethylamine (270 mg) were added thereto. ,2.7mmol), reacted at room temperature for 1.5h. Add water (50mL) to quench the reaction, and extract the aqueous phase with dichloromethane (35mL*3). After the organic phases were combined, they were dried over anhydrous sodium sulfate, filtered, and the solvent was spin-dried. The crude product was purified by column chromatography (eluent: dichloromethane: methanol = 15:1) to obtain compound D-4 (127 mg) as a white solid.
ESI-MS[M+H]+:619.3.ESI-MS[M+H] + :619.3.
1H NMR(400MHz,DMSO-d6)δ11.00(s,1H),9.21(d,J＝4.2Hz,1H),8.14(d,J＝7.9Hz,1H),8.07(s,1H),8.02-7.98(m,1H),7.92-7.88(m,1H),7.83-7.73(m,2H),7.57-7.45(m,2H),7.43-7.27(m,4H),7.24-7.19(m,2H),5.13(dd,J＝13.2,5.0Hz,1H),4.62(s,1H),4.53-4.32(m,5H),4.16(s,1H),3.96(s,1H),3.53-3.44(m,1H),3.30-3.13(m,3H),3.00-2.84(m,1H),2.66-2.55(m,1H),2.47-2.27(m,1H),2.05-1.93(m,1H). 1 H NMR (400MHz, DMSO-d 6 ) δ11.00 (s, 1H), 9.21 (d, J = 4.2Hz, 1H), 8.14 (d, J = 7.9Hz, 1H), 8.07 (s, 1H) ,8.02-7.98(m,1H),7.92-7.88(m,1H),7.83-7.73(m,2H),7.57-7.45(m,2H),7.43-7.27(m,4H),7.24-7.19( m,2H),5.13(dd,J＝13.2,5.0Hz,1H),4.62(s,1H),4.53-4.32(m,5H),4.16(s,1H),3.96(s,1H),3.53 -3.44(m,1H),3.30-3.13(m,3H),3.00-2.84(m,1H),2.66-2.55(m,1H),2.47-2.27(m,1H),2.05-1.93(m, 1H).
实施例1.4-A：N-(4-((N-苄基-2-羟基乙酰胺基)甲基)苄基)-2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-甲酰胺(D-4-A)的合成
Example 1.4-A: N-(4-((N-benzyl-2-hydroxyacetamido)methyl)benzyl)-2-(2,6-dioxopiperidin-3-yl)- Synthesis of 1-oxoisoindoline-5-carboxamide (D-4-A)
除在步骤一中采用苯甲醇替代2-(1-萘基)乙醇外，参考D-4的合成方法，得到D-4-A。 Except using benzyl alcohol instead of 2-(1-naphthyl)ethanol in step 1, refer to the synthesis method of D-4 to obtain D-4-A.
1H NMR(400MHz,DMSO)δ11.01(s,1H),9.23(t,J＝6.0Hz,1H),8.10(s,1H),8.02(d,J＝8.0Hz,1H),7.82(d,J＝7.9Hz,1H),7.42–7.25(m,5H),7.22-7.15(m,4H),5.14(dd,J＝13.3,5.1Hz,1H),4.75(br s,1H),4.59–4.32(m,8H),4.17(d,J＝3.6Hz,2H),2.99–2.84(m,1H),2.66–2.54(m,1H),2.47–2.32(m,1H),2.07-1.97(m,1H).ESI-MS(M+H)+:555.3 1 H NMR (400MHz, DMSO) δ11.01(s,1H),9.23(t,J＝6.0Hz,1H),8.10(s,1H),8.02(d,J＝8.0Hz,1H),7.82( d,J＝7.9Hz,1H),7.42–7.25(m,5H),7.22-7.15(m,4H),5.14(dd,J＝13.3,5.1Hz,1H),4.75(br s,1H), 4.59–4.32(m,8H),4.17(d,J＝3.6Hz,2H),2.99–2.84(m,1H),2.66–2.54(m,1H),2.47–2.32(m,1H),2.07- 1.97(m,1H).ESI-MS(M+H) + :555.3
实施例1.4-B：2-(2,6-二氧代哌啶-3-基)-N-(4-((3-羟基-N-苯乙基丙酰胺基)甲基)苄基)-1-氧代异吲哚啉-5-甲酰胺(D-4-B)的合成
Example 1.4-B: 2-(2,6-dioxopiperidin-3-yl)-N-(4-((3-hydroxy-N-phenylethylpropionamido)methyl)benzyl) -Synthesis of 1-oxoisoindoline-5-carboxamide (D-4-B)
除在步骤一中采用苯乙醇替代2-(1-萘基)乙醇；和步骤三中采用羟基丙酸代替羟基乙酸外，参考D-4的合成方法，得到D-4-B。Except that phenylethanol is used instead of 2-(1-naphthyl)ethanol in step one; and hydroxypropionic acid is used instead of glycolic acid in step three, D-4-B is obtained by referring to the synthesis method of D-4.
1H NMR(400MHz,DMSO)δ11.00(s,1H),9.20(dd,J＝13.9,6.0Hz,1H),8.09(s,1H),8.00(d,J＝7.9Hz,1H),7.81(d,J＝7.9Hz,1H),7.34-7.24(m,4H),7.23-7.15(m,5H),5.14(dd,J＝13.3,5.1Hz,1H),4.56–4.35(m,7H),3.69–3.57(m,2H),3.42(t,J＝7.5Hz,2H),2.98–2.85(m,1H),2.81(t,J＝7.6Hz,1H),2.78–2.69(m,1H),2.66-2.54(m,1H),2.48-2.34(m,3H),2.07–1.96(m,1H).ESI-MS(M+H)+:583.4. 1 H NMR (400MHz, DMSO) δ11.00(s,1H),9.20(dd,J＝13.9,6.0Hz,1H),8.09(s,1H),8.00(d,J＝7.9Hz,1H), 7.81(d,J＝7.9Hz,1H),7.34-7.24(m,4H),7.23-7.15(m,5H),5.14(dd,J＝13.3,5.1Hz,1H),4.56–4.35(m, 7H),3.69–3.57(m,2H),3.42(t,J＝7.5Hz,2H),2.98–2.85(m,1H),2.81(t,J＝7.6Hz,1H),2.78–2.69(m ,1H),2.66-2.54(m,1H),2.48-2.34(m,3H),2.07–1.96(m,1H).ESI-MS(M+H) + :583.4.
实施例1.4-C：2-(2,6-二氧代哌啶-3-基)-N-(4-((2-巯基-N-(2-(萘-1-基)乙基)乙酰氨基)甲基)苄基)-1-氧代异吲哚啉-5-甲酰胺(D-4-C)的合成
Example 1.4-C: 2-(2,6-dioxopiperidin-3-yl)-N-(4-((2-mercapto-N-(2-(naphthyl-1-yl)ethyl)) Synthesis of acetamido)methyl)benzyl)-1-oxoisoindoline-5-carboxamide (D-4-C)
除在步骤三中采用S-乙酰硫基乙酸替代羟基乙酸外，参考D-4的合成方法，得到D-4-C-1；Except using S-acetylthioacetic acid instead of glycolic acid in step 3, refer to the synthesis method of D-4 to obtain D-4-C-1;
由D-4-C-1制备得到D-4-C，具体反应方程式和操作步骤如下：
D-4-C is prepared from D-4-C-1. The specific reaction equation and operation steps are as follows:
将化合物D-4-C-1(50mg,0.074mmol)溶于5mL四氢呋喃中，向其中加入稀盐酸(1M，1mL)。50℃反应过夜后，TLC显示反应毕。向反应液加入aq.NaHCO3(10mL)后，用二氯甲烷萃取(10mL*3)。有机相合并干燥后，旋干溶剂。粗品经柱层析纯化(洗脱剂为二氯甲烷：甲醇＝15:1)，得化合物D-4-C(1mg,产率2％)。1H NMR(400MHz,DMSO)δ11.02(s,1H),9.23(t,J＝6.0Hz,1H),8.16–7.96(m,3H),7.94-7.87(m,1H),7.84-7.74(m,2H),7.56–7.26(m,6H),7.26–7.17(m,2H),5.14(dd,J＝13.3,5.1Hz,1H),4.58(d,J＝9.9Hz,2H),4.53–4.32(m,4H),3.57–3.43(m,2H),3.33–3.28(m,2H),3.22–3.14(m,1H),2.98–2.77(m,2H),2.65-2.56(m,1H),2.43-2.36(m,1H),2.05-1.95(m,1H).ESI-MS(M+H)+:635.4。Compound D-4-C-1 (50 mg, 0.074 mmol) was dissolved in 5 mL of tetrahydrofuran, and dilute hydrochloric acid (1 M, 1 mL) was added thereto. After reacting at 50°C overnight, TLC showed that the reaction was complete. After adding aq.NaHCO 3 (10 mL) to the reaction solution, the mixture was extracted with dichloromethane (10 mL*3). After the organic phases were combined and dried, the solvent was spun off. The crude product was purified by column chromatography (eluent: dichloromethane: methanol = 15:1) to obtain compound D-4-C (1 mg, yield 2%). 1 H NMR (400MHz, DMSO) δ11.02 (s, 1H), 9.23 (t, J = 6.0Hz, 1H), 8.16–7.96 (m, 3H), 7.94-7.87 (m, 1H), 7.84-7.74 (m,2H),7.56–7.26(m,6H),7.26–7.17(m,2H),5.14(dd,J＝13.3,5.1Hz,1H),4.58(d,J＝9.9Hz,2H), 4.53–4.32(m,4H),3.57–3.43(m,2H),3.33–3.28(m,2H),3.22–3.14(m,1H),2.98–2.77(m,2H),2.65-2.56(m ,1H),2.43-2.36(m,1H),2.05-1.95(m,1H).ESI-MS(M+H) + :635.4.
实施例1.5：2-(2,6-二氧代哌啶-3-基)-N-(4-((2-(1-(羟基甲基)-9H-咔唑-9-基)乙酰胺)甲基)苄基)-1-氧代异吲哚啉-5-甲酰胺(D-5)的合成
Example 1.5: 2-(2,6-dioxopiperidin-3-yl)-N-(4-((2-(1-(hydroxymethyl)-9H-carbazol-9-yl)ethyl) Synthesis of amide)methyl)benzyl)-1-oxoisoindoline-5-carboxamide (D-5)
步骤一：D-5-1的合成Step 1: Synthesis of D-5-1
将1-溴咔唑(1.2g,4.9mmol)加到50mL THF中，冰浴下，向其中加入NaH(590mg,14.6mmol，60％)，加毕常温反应1h，加入溴乙酸叔丁酯(1.14g,5.9mmol)。常温反应过夜。加水(60mL)淬灭反应。水相用乙酸乙酯萃取(80mL*3)。有机相合并后用饱和食盐水(20mL)洗涤，再无水硫酸钠干燥并旋干溶剂。残余物经硅胶柱层析分离(流动相为石油醚：乙酸乙酯＝10：1)得到化合物D-5-1(700mg)。Add 1-bromocarbazole (1.2g, 4.9mmol) to 50mL THF, add NaH (590mg, 14.6mmol, 60%) under ice bath, complete the addition and react at room temperature for 1h, add tert-butyl bromoacetate ( 1.14g, 5.9mmol). Reaction at room temperature overnight. The reaction was quenched by adding water (60 mL). The aqueous phase was extracted with ethyl acetate (80mL*3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and the solvent was spun off. The residue was separated by silica gel column chromatography (mobile phase was petroleum ether: ethyl acetate = 10:1) to obtain compound D-5-1 (700 mg).
步骤二：D-5-2的合成Step 2: Synthesis of D-5-2
将化合物D-5-1(700mg,1.9mmol)溶于15mL二氯甲烷中，向其中加入5mL三氟乙酸。常温反应1h。旋干溶液，得化合物D-5-2(700mg,粗品)直接用于下一步反应。Compound D-5-1 (700 mg, 1.9 mmol) was dissolved in 15 mL of methylene chloride, and 5 mL of trifluoroacetic acid was added thereto. Reaction at room temperature for 1 hour. The solution was spin-dried to obtain compound D-5-2 (700 mg, crude product), which was directly used in the next reaction.
步骤三：D-5-3的合成Step 3: Synthesis of D-5-3
将化合物D-5-2(700mg,1.9mmol)溶于40mL DCM中，向其中加入1-(N-Boc-氨基甲基)-4-(氨基甲基)苯(560mg,2.4mmol)，HATU(1.1g,2.9mmol) 和三乙胺(800mg,7.9mmol)。常温反应过夜。加水(20mL)淬灭反应后，用二氯甲烷萃取(80mL*3)。有机相合并后，用饱和食盐水(20mL)洗涤再用无水硫酸钠干燥并旋干溶剂。残余物经硅胶柱层析纯化(流动相为石油醚：乙酸乙酯＝1：1)得到化合物D-5-3(800mg)。Compound D-5-2 (700 mg, 1.9 mmol) was dissolved in 40 mL DCM, and 1-(N-Boc-aminomethyl)-4-(aminomethyl)benzene (560 mg, 2.4 mmol) was added thereto, HATU (1.1g,2.9mmol) and triethylamine (800 mg, 7.9 mmol). Reaction at room temperature overnight. After adding water (20 mL) to quench the reaction, extract with dichloromethane (80 mL*3). After the organic phases were combined, they were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and the solvent was spun dry. The residue was purified by silica gel column chromatography (mobile phase was petroleum ether: ethyl acetate = 1:1) to obtain compound D-5-3 (800 mg).
1H NMR(400MHz,DMSO-d6)δ8.66(t,J＝6.0Hz,1H),8.22-8.19(m,2H),7.59(dd,J＝11.2,7.7Hz,2H),7.52-7.45(m,1H),7.36(s,1H),7.29-7.25(m,1H),7.22-7.11(m,5H),5.47(s,2H),4.28(d,J＝5.8Hz,2H),4.09(d,J＝5.8Hz,2H),1.39(s,9H). 1 H NMR (400MHz, DMSO-d 6 ) δ8.66 (t, J = 6.0 Hz, 1H), 8.22-8.19 (m, 2H), 7.59 (dd, J = 11.2, 7.7 Hz, 2H), 7.52- 7.45(m,1H),7.36(s,1H),7.29-7.25(m,1H),7.22-7.11(m,5H),5.47(s,2H),4.28(d,J＝5.8Hz,2H) ,4.09(d,J=5.8Hz,2H),1.39(s,9H).
步骤四：D-5-4的合成Step 4: Synthesis of D-5-4
将化合物D-5-3(500mg,0.9mmol)，氰化锌(230mg,1.9mmol)，Pd2(dba)3(170mg,0.2mmol)和DPPF(200mg,0.4mmol)溶于18mL DMF中。氮气保护下，120℃反应过夜。加水(60mL)淬灭反应后，用乙酸乙酯萃取(60mL*3)。有机相合并后用饱和食盐水(20mL)洗涤，再用无水硫酸钠干燥并旋干溶剂。残余物经硅胶柱层析纯化(流动相为二氯甲烷：甲醇＝20：1)得到化合物D-5-4(370mg)。Compound D-5-3 (500 mg, 0.9 mmol), zinc cyanide (230 mg, 1.9 mmol), Pd 2 (dba) 3 (170 mg, 0.2 mmol) and DPPF (200 mg, 0.4 mmol) were dissolved in 18 mL DMF. Under nitrogen protection, the reaction was carried out at 120°C overnight. After adding water (60 mL) to quench the reaction, extract with ethyl acetate (60 mL*3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and the solvent was spun off. The residue was purified by silica gel column chromatography (the mobile phase was dichloromethane: methanol = 20:1) to obtain compound D-5-4 (370 mg).
1H NMR(400MHz,DMSO-d6)δ8.82(t,J＝5.7Hz,1H),8.55(d,J＝7.7Hz,1H),8.29(d,J＝7.7Hz,1H),7.87(d,J＝7.4Hz,1H),7.64(d,J＝8.3Hz,1H),7.57-7.53(m,1H),7.41-7.32(m,3H),7.24(d,J＝8.1Hz,2H),7.17(d,J＝8.1Hz,2H),5.43(s,2H),4.31(d,J＝5.6Hz,2H),4.09(d,J＝6.1Hz,2H),1.36(s,9H). 1 H NMR (400MHz, DMSO-d 6 ) δ8.82(t,J＝5.7Hz,1H),8.55(d,J＝7.7Hz,1H),8.29(d,J＝7.7Hz,1H),7.87 (d,J＝7.4Hz,1H),7.64(d,J＝8.3Hz,1H),7.57-7.53(m,1H),7.41-7.32(m,3H),7.24(d,J＝8.1Hz, 2H),7.17(d,J＝8.1Hz,2H),5.43(s,2H),4.31(d,J＝5.6Hz,2H),4.09(d,J＝6.1Hz,2H),1.36(s, 9H).
步骤五：D-5-5的合成Step 5: Synthesis of D-5-5
将化合物D-5-4(300mg,0.6mmol)溶于10mL吡啶和3mL AcOH和3mL水中。向其中加入次磷酸钠(650mg,6.0mmol)和雷尼镍(1g)。常温反应过夜。将反应液过滤，滤液加饱和碳酸氢钠(40mL)后，用乙酸乙酯萃取(80mL*3)。有机相合并后用饱和食盐水(20mL*2)洗涤，再用无水硫酸钠干燥并旋干溶剂，得到化合物D-5-5(350mg)，直接用于下一步反应。Compound D-5-4 (300 mg, 0.6 mmol) was dissolved in 10 mL pyridine and 3 mL AcOH and 3 mL water. Sodium hypophosphite (650 mg, 6.0 mmol) and Raney Nickel (1 g) were added thereto. Reaction at room temperature overnight. The reaction solution was filtered, and saturated sodium bicarbonate (40 mL) was added to the filtrate, and then extracted with ethyl acetate (80 mL*3). The organic phases were combined, washed with saturated brine (20 mL*2), dried over anhydrous sodium sulfate, and spin-dried to obtain compound D-5-5 (350 mg), which was directly used in the next reaction.
ESI-MS[M+H]+:472.3.ESI-MS[M+H] + :472.3.
步骤六：D-5-6的合成Step 6: Synthesis of D-5-6
将化合物D-5-5(350mg,0.6mmol)溶于25mL甲醇中，冰浴下向其中加入硼氢化钠(40mg,1.1mmol)。常温反应2h。加水(20mL)淬灭反应，用二 氯甲烷萃取(60mL*3)。有机相合并后用饱和食盐水(20mL)洗涤，再用无水硫酸钠干燥并旋干溶剂。残余物经硅胶柱层析纯化(流动相为二氯甲烷：甲醇＝20：1)，得到化合物D-5-6(80mg)。Compound D-5-5 (350 mg, 0.6 mmol) was dissolved in 25 mL of methanol, and sodium borohydride (40 mg, 1.1 mmol) was added thereto under ice bath. Reaction at room temperature for 2 hours. Add water (20 mL) to quench the reaction, and use 2 Methyl chloride extraction (60mL*3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and the solvent was spun off. The residue was purified by silica gel column chromatography (the mobile phase was dichloromethane: methanol = 20:1) to obtain compound D-5-6 (80 mg).
ESI-MS[M-18+H]+:456.3.ESI-MS[M-18+H] + :456.3.
步骤七：D-5-7的合成Step 7: Synthesis of D-5-7
将化合物D-5-6(80mg,0.17mmol)溶于5mL二氯甲烷中，向其中加入HCl/dioxane(3mL,4M)。常温反应1h。旋干溶剂得到化合物D-5-7(80mg,粗品)，直接用于下一步反应。Compound D-5-6 (80 mg, 0.17 mmol) was dissolved in 5 mL of dichloromethane, and HCl/dioxane (3 mL, 4M) was added thereto. Reaction at room temperature for 1 hour. The solvent was spun dry to obtain compound D-5-7 (80 mg, crude product), which was directly used in the next reaction.
步骤八：D-5的合成Step 8: Synthesis of D-5
将化合物D-5-7溶于15mL DCM中，向其中加入化合物D-1-3(50mg,0.17mmol)，HATU(90mg,0.24mmol)和三乙胺(65mg,0.6mmol)。常温反应过夜后，TLC显示反应毕。加水(20mL)淬灭反应，水相用二氯甲烷萃取(80mL*3)。有机相合并后用饱和食盐水(20mL)洗涤，再用无水硫酸钠干燥并旋干溶剂。粗品用柱层析纯化(流动相为二氯甲烷：甲醇＝10：1)得化合物D-5-8(24mg)。Compound D-5-7 was dissolved in 15 mL DCM, and compound D-1-3 (50 mg, 0.17 mmol), HATU (90 mg, 0.24 mmol) and triethylamine (65 mg, 0.6 mmol) were added thereto. After reacting at room temperature overnight, TLC showed that the reaction was complete. Add water (20 mL) to quench the reaction, and extract the aqueous phase with dichloromethane (80 mL*3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and the solvent was spun off. The crude product was purified by column chromatography (the mobile phase was dichloromethane: methanol = 10:1) to obtain compound D-5-8 (24 mg).
ESI-MS[M-18+H]+:626.3.ESI-MS[M-18+H] + :626.3.
1H NMR(400MHz,DMSO-d6)δ11.01(s,1H),9.22(t,J＝5.9Hz,1H),8.71(t,J＝5.9Hz,1H),8.18-8.10(m,2H),8.08(s,1H),8.00(d,J＝8.0Hz,1H),7.82(d,J＝7.9Hz,1H),7.48(d,J＝8.2Hz,1H),7.45-7.40(m,1H),7.33(d,J＝6.6Hz,1H),7.28(d,J＝8.1Hz,2H),7.24-7.18(m,3H),7.17-7.12(m,1H),5.55(t,J＝5.3Hz,1H),5.41(s,2H),5.13(dd,J＝13.3,5.1Hz,1H),4.77(d,J＝5.3Hz,2H),4.55-4.37(m,4H),4.28(d,J＝5.8Hz,2H),2.99-2.85(m,1H),2.68-2.56(m,1H),2.46-2.32(m,1H),2.12-1.99(m,1H). 1 H NMR (400MHz, DMSO-d 6 ) δ11.01 (s, 1H), 9.22 (t, J = 5.9 Hz, 1H), 8.71 (t, J = 5.9 Hz, 1H), 8.18-8.10 (m, 2H),8.08(s,1H),8.00(d,J＝8.0Hz,1H),7.82(d,J＝7.9Hz,1H),7.48(d,J＝8.2Hz,1H),7.45-7.40( m,1H),7.33(d,J＝6.6Hz,1H),7.28(d,J＝8.1Hz,2H),7.24-7.18(m,3H),7.17-7.12(m,1H),5.55(t ,J＝5.3Hz,1H),5.41(s,2H),5.13(dd,J＝13.3,5.1Hz,1H),4.77(d,J＝5.3Hz,2H),4.55-4.37(m,4H) ,4.28(d,J=5.8Hz,2H),2.99-2.85(m,1H),2.68-2.56(m,1H),2.46-2.32(m,1H),2.12-1.99(m,1H).
实施例1.6：N-(4-((2-(9H-吡啶并[2,3-b]吲哚-9-基)乙酰胺)甲基)-2-(2-羟基乙基氧基)苄基)-2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-甲酰胺(D-6)的合成
Example 1.6: N-(4-((2-(9H-pyrido[2,3-b]indol-9-yl)acetamide)methyl)-2-(2-hydroxyethyloxy) Synthesis of benzyl)-2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-carboxamide (D-6)
步骤一：D-6-1的合成Step 1: Synthesis of D-6-1
将3-羟基-4-甲基苯腈(2.66g，20mmol)溶解于DMF(30ml)中，向其中加入溴乙醇(3.75g，30mmol)和碳酸钾(8.28g，60mmol)。80℃反应3h。反应液降至室温，加入水(300ml)后，用乙酸乙酯萃取(100mL*2)。合并有机相后，饱和食盐水洗涤(50mL*2)。收集有机相，干燥，过滤，旋干溶剂得D-6-1(2.55g)。3-Hydroxy-4-methylbenzonitrile (2.66g, 20mmol) was dissolved in DMF (30ml), and bromoethanol (3.75g, 30mmol) and potassium carbonate (8.28g, 60mmol) were added thereto. React at 80°C for 3 hours. The reaction solution was cooled to room temperature, water (300 ml) was added, and extracted with ethyl acetate (100 mL*2). After combining the organic phases, wash with saturated brine (50mL*2). The organic phase was collected, dried, filtered, and the solvent was spin-dried to obtain D-6-1 (2.55g).
1H NMR(400MHz,DMSO-d6)7.37(s,1H),7.34-7.29(m,2H),4.88(t,J＝4.0Hz,1H),4.07(t,J＝4.0Hz,2H),2.23(s,3H). 1 H NMR (400MHz, DMSO-d 6 )7.37(s,1H),7.34-7.29(m,2H),4.88(t,J＝4.0Hz,1H),4.07(t,J＝4.0Hz,2H) ,2.23(s,3H).
步骤二：D-6-2的合成Step 2: Synthesis of D-6-2
将化合物D-6-1(2.55g，14.4mmol)溶解于DCM(30ml)中，向其中加入TEA(2.91g，28.8mmol)。冰浴下缓慢加入乙酰氯(1.36g，17.3mmol)，加料完毕温度上升到20℃。继续反应3h。倒入冰水(100ml)中，乙酸乙酯萃 取(100mL*2)。合并有机相，饱和食盐水洗(50mL*2)。收集有机相，干燥，过滤，旋干溶剂。粗品用柱层析纯化(石油醚：乙酸乙酯＝5：1)得到D-6-2(2.65g)。Compound D-6-1 (2.55g, 14.4mmol) was dissolved in DCM (30ml), and TEA (2.91g, 28.8mmol) was added thereto. Acetyl chloride (1.36g, 17.3mmol) was slowly added under ice bath, and the temperature rose to 20°C after the addition was completed. Continue the reaction for 3 hours. Pour into ice water (100ml), extract with ethyl acetate Take (100mL*2). Combine the organic phases and wash with saturated brine (50mL*2). The organic phase was collected, dried, filtered, and the solvent was spun off. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain D-6-2 (2.65g).
步骤三：D-6-3的合成Step 3: Synthesis of D-6-3
将化合物D-6-2(2.65g,12.1mmol)溶解于四氯化碳(40ml)中，向其中加入NBS(2.15g,12.1mmol)和AIBN(0.40g，2.42mmol)。80℃反应3h。将反应液旋干，粗品用柱层析纯化(石油醚:乙酸乙酯＝4:1)得到D-6-3(2.25g)。Compound D-6-2 (2.65g, 12.1mmol) was dissolved in carbon tetrachloride (40ml), and NBS (2.15g, 12.1mmol) and AIBN (0.40g, 2.42mmol) were added thereto. React at 80°C for 3 hours. The reaction liquid was spun to dryness, and the crude product was purified by column chromatography (petroleum ether:ethyl acetate=4:1) to obtain D-6-3 (2.25g).
步骤四：D-6-4的合成Step 4: Synthesis of D-6-4
将化合物D-6-3(1.49g，5.0mmol)和NaN3(0.325g，5.0mmol)溶解于DMF(15ml)。室温反应3h。加水(50ml)淬灭反应。水相用乙酸乙酯(100mL*2)萃取后，合并有机相。有机相再用饱和食盐水洗(50mL*2)。收集有机相，干燥，过滤后旋干溶剂得化合物D-6-4(1.3g)。Compound D-6-3 (1.49g, 5.0mmol) and NaN 3 (0.325g, 5.0mmol) were dissolved in DMF (15ml). React at room temperature for 3 hours. The reaction was quenched by adding water (50 ml). After the aqueous phase was extracted with ethyl acetate (100mL*2), the organic phases were combined. Wash the organic phase with saturated brine (50mL*2). The organic phase was collected, dried, filtered and the solvent was spin-dried to obtain compound D-6-4 (1.3g).
步骤五：D-6-5的合成Step 5: Synthesis of D-6-5
将化合物D-6-4(1.3g，5.0mmol)，三苯基膦(1.31g，5.0mmol)溶解于THF/H2O(20ml/5ml)中。室温反应5h。加入稀盐酸(1M，50ml)淬灭反应。水相用乙酸乙酯萃取(50ml*2)后，水相再用饱和碳酸钠溶液调节pH至10。水相再用乙酸乙酯萃取(50ml*3)。合并有机相，干燥，过滤后旋干溶剂得化合物D-6-5(1.3g，产率100％)。Compound D-6-4 (1.3g, 5.0mmol) and triphenylphosphine (1.31g, 5.0mmol) were dissolved in THF/H 2 O (20ml/5ml). React at room temperature for 5 hours. Dilute hydrochloric acid (1M, 50ml) was added to quench the reaction. After the aqueous phase was extracted with ethyl acetate (50ml*2), the pH of the aqueous phase was adjusted to 10 with saturated sodium carbonate solution. The aqueous phase was extracted with ethyl acetate (50ml*3). The organic phases were combined, dried, filtered, and the solvent was spin-dried to obtain compound D-6-5 (1.3 g, yield 100%).
ESI-MS(M+H)+:235.2.ESI-MS(M+H) + :235.2.
步骤六：D-6-6的合成Step 6: Synthesis of D-6-6
将化合物D-6-5(1.17g，5.0mmol)溶解于乙酸乙酯(50ml)，向其中加入饱和碳酸氢钠溶液(20ml)。冰浴下向其中后加入FmocCl(2.1g，7.5mmol)。室温反应3h。加入水(50ml)淬灭反应。水相用乙酸乙酯萃取(100ml*2)后，合并有机相。干燥，过滤，旋干溶剂。粗品用柱层析纯化(石油醚：乙酸乙酯＝1:1)得到化合物D-6-6(1.3g，57％)。Compound D-6-5 (1.17g, 5.0mmol) was dissolved in ethyl acetate (50ml), and saturated sodium bicarbonate solution (20ml) was added thereto. FmocCl (2.1 g, 7.5 mmol) was added thereto under ice bath. React at room temperature for 3 hours. Water (50 ml) was added to quench the reaction. After the aqueous phase was extracted with ethyl acetate (100ml*2), the organic phases were combined. Dry, filter, and spin down the solvent. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain compound D-6-6 (1.3 g, 57%).
ESI-MS[M+H]+:457.2.ESI-MS[M+H] + :457.2.
步骤七：D-6-7的合成 Step 7: Synthesis of D-6-7
将化合物D-6-6(1.3g，2.85mmol)溶解于40ml甲醇中，向其中加入(Boc)2O(1.24g，5.7mmol)和雷尼镍(2.0g)。在氢气氛围下室温反应2h。将反应液过滤，滤液旋干得化合物D-6-7(1.15g)。Compound D-6-6 (1.3 g, 2.85 mmol) was dissolved in 40 ml of methanol, and (Boc) 2 O (1.24 g, 5.7 mmol) and Raney Nickel (2.0 g) were added thereto. React at room temperature for 2 hours under hydrogen atmosphere. The reaction solution was filtered, and the filtrate was spun to dryness to obtain compound D-6-7 (1.15g).
ESI-MS[M+H]+:561.2.ESI-MS[M+H] + :561.2.
步骤八：D-6-8的合成Step 8: Synthesis of D-6-8
将化合物D-6-7(1.15g，2.1mmol)溶解于20ml二氯甲烷中，向其中加入三氟乙酸(4ml)。室温反应2h。旋干溶剂，得化合物D-6-8(1.3g,粗品)，直接用于下一步反应。Compound D-6-7 (1.15g, 2.1mmol) was dissolved in 20ml of methylene chloride, and trifluoroacetic acid (4ml) was added thereto. React at room temperature for 2 hours. The solvent was spun dry to obtain compound D-6-8 (1.3g, crude product), which was directly used in the next reaction.
步骤九：D-6-10的合成Step 9: Synthesis of D-6-10
将化合物D-6-8(0.97g，2.1mmol),化合物D-6-9(475mg,2.1mmol),DIPEA(0.81g,6.3mmol)溶解于DMF(20ml)中。冰浴下缓慢加入HATU(0.96g,2.52mmol)。加毕常温反应2h。加入冰水(100ml)淬灭反应。水相用乙酸乙酯萃取(200mL*2)。有机相合并后，用饱和食盐水再洗(50mL*2)。收集有机相，干燥，过滤，旋干溶剂。粗品用柱层析(石油醚：乙酸乙酯＝1：1)得到化合物D-6-10(1.05g)。Compound D-6-8 (0.97g, 2.1mmol), compound D-6-9 (475mg, 2.1mmol), DIPEA (0.81g, 6.3mmol) were dissolved in DMF (20ml). HATU (0.96g, 2.52mmol) was slowly added under ice bath. After the addition, react at room temperature for 2 hours. Ice water (100 ml) was added to quench the reaction. The aqueous phase was extracted with ethyl acetate (200mL*2). After combining the organic phases, wash with saturated brine (50mL*2). The organic phase was collected, dried, filtered, and the solvent was spun off. The crude product was subjected to column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain compound D-6-10 (1.05 g).
ESI-MS[M+H]+:669.3.ESI-MS[M+H] + :669.3.
步骤十：D-6-11的合成Step 10: Synthesis of D-6-11
将化合物D-6-10(335mg，0.5mmol)溶解于30mL甲醇中，在冰浴下缓慢加入氢氧化锂水溶液(5ml,2M)。常温反应16h。加入水(100ml)稀释。水相用乙酸乙酯萃取(100mL*3)后合并有机相。再用饱和食盐水再洗(50mL*2)。收集有机相，干燥，过滤，旋干溶剂。粗品用柱层析纯化(石油醚：乙酸乙酯＝1：1)得到化合物D-6-11(202mg)。Compound D-6-10 (335 mg, 0.5 mmol) was dissolved in 30 mL of methanol, and an aqueous lithium hydroxide solution (5 ml, 2 M) was slowly added under an ice bath. Reaction at room temperature for 16 hours. Add water (100ml) to dilute. The aqueous phase was extracted with ethyl acetate (100mL*3) and the organic phases were combined. Then wash with saturated saline (50mL*2). The organic phase was collected, dried, filtered, and the solvent was spun off. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain compound D-6-11 (202 mg).
步骤十一：化合物D-6的合成Step 11: Synthesis of Compound D-6
将中间体D-6-11(202mg，0.5mmol),D-1-3(144mg,0.5mmol),DIPEA(194mg,1.5mmol)溶解于10mL DMF中。冰浴下缓慢加入HATU(228mg,0.6mmol)。常温反应2h。加入冰水(100ml)淬灭反应。水相用乙酸乙酯萃取(200mL*2)。有机相合并后，用饱和食盐水再洗(50mL*2)。收集有机相，干燥，过滤，旋干溶剂。粗品用柱层析纯化(二氯甲烷：甲醇＝15：1)得到化合物D-6(22mg,产率7％)。Dissolve intermediates D-6-11 (202 mg, 0.5 mmol), D-1-3 (144 mg, 0.5 mmol), and DIPEA (194 mg, 1.5 mmol) in 10 mL DMF. HATU (228 mg, 0.6 mmol) was slowly added under ice bath. Reaction at room temperature for 2 hours. Ice water (100 ml) was added to quench the reaction. The aqueous phase was extracted with ethyl acetate (200mL*2). After combining the organic phases, wash with saturated brine (50mL*2). The organic phase was collected, dried, filtered, and the solvent was spun off. The crude product was purified by column chromatography (dichloromethane: methanol = 15:1) to obtain compound D-6 (22 mg, yield 7%).
ESI-MS[M+H]+:675.3. ESI-MS[M+H] + :675.3.
1H NMR(400MHz,DMSO-d6)δ11.01(s,1H),8.94(t,J＝5.8Hz,1H),8.72(t,J＝5.9Hz,1H),8.55(dd,J＝7.6,1.4Hz,1H),8.44(dd,J＝4.8,1.4Hz,1H),8.21(d,J＝7.7Hz,1H),8.09(s,1H),8.00(d,J＝8.1Hz,1H),7.82(d,J＝7.9Hz,1H),7.58(d,J＝8.2Hz,1H),7.49(t,J＝7.7Hz,1H),7.32-7.23(m,2H),7.16(d,J＝7.7Hz,1H),6.87(s,1H),6.82(d,J＝7.8Hz,1H),5.20-5.10(m,3H),4.94(t,J＝5.7Hz,1H),4.47(dt,J＝41.6,17.7Hz,4H),4.28(d,J＝5.8Hz,2H),3.99(t,J＝4.9Hz,2H),3.76(dd,J＝10.2,5.2Hz,2H),2.99-2.85(m,1H),2.65-2.56(m,1H),2.47-2.36(m,1H),2.06-1.95(m,1H). 1 H NMR (400MHz, DMSO-d 6 ) δ11.01 (s, 1H), 8.94 (t, J = 5.8Hz, 1H), 8.72 (t, J = 5.9Hz, 1H), 8.55 (dd, J = 7.6,1.4Hz,1H),8.44(dd,J＝4.8,1.4Hz,1H),8.21(d,J＝7.7Hz,1H),8.09(s,1H),8.00(d,J＝8.1Hz, 1H),7.82(d,J＝7.9Hz,1H),7.58(d,J＝8.2Hz,1H),7.49(t,J＝7.7Hz,1H),7.32-7.23(m,2H),7.16( d,J＝7.7Hz,1H),6.87(s,1H),6.82(d,J＝7.8Hz,1H),5.20-5.10(m,3H),4.94(t,J＝5.7Hz,1H), 4.47(dt,J＝41.6,17.7Hz,4H),4.28(d,J＝5.8Hz,2H),3.99(t,J＝4.9Hz,2H),3.76(dd,J＝10.2,5.2Hz,2H ),2.99-2.85(m,1H),2.65-2.56(m,1H),2.47-2.36(m,1H),2.06-1.95(m,1H).
实施例1.7 4-((4-(3-氯-4-羟基苯基)哌嗪-1-基)甲基)-N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)苯甲酰胺(D-7)的合成
Example 1.7 4-((4-(3-chloro-4-hydroxyphenyl)piperazin-1-yl)methyl)-N-((2-(2,6-dioxopiperidine-3- Synthesis of 1-oxoisoindolin-5-yl)methyl)benzamide (D-7)
步骤一：D-7-1的合成Step 1: Synthesis of D-7-1
将4-溴-2-氯苯酚(600mg,2.9mmol)和N-Boc-哌嗪(680mg,3.6mmol)溶于30mL甲苯中，向其中加入LiHMDS(6.6mL,1M)，Pd2(dba)3(120mg,0.15mmol)和2,8,9-三异丁基-2,5,8,9-四氮杂-1-磷酸双向环[3.3.3]十一烷(120mg,0.3mmol)。氮气保护下80℃反应过夜。加水(60mL)淬灭反应，水相用乙酸乙酯萃取(60mL*3)。合并有机相后，用饱和食盐水(20mL)洗涤。有机相用无水硫酸钠干燥并浓缩。粗品用层析柱纯化(流动相为石油醚：乙酸乙酯＝4：1)得到化合物D-7-1(270mg,产率30％)。Dissolve 4-bromo-2-chlorophenol (600mg, 2.9mmol) and N-Boc-piperazine (680mg, 3.6mmol) in 30mL toluene, add LiHMDS (6.6mL, 1M), Pd 2 (dba) 3 (120mg, 0.15mmol) and 2,8,9-triisobutyl-2,5,8,9-tetraaza-1-phosphate bidirectional cyclo[3.3.3]undecane (120mg, 0.3mmol) . The reaction was carried out at 80°C overnight under nitrogen protection. Water (60 mL) was added to quench the reaction, and the aqueous phase was extracted with ethyl acetate (60 mL*3). The organic phases were combined and washed with saturated brine (20 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated. The crude product was purified using a chromatography column (the mobile phase was petroleum ether: ethyl acetate = 4:1) to obtain compound D-7-1 (270 mg, yield 30%).
ESI-MS[M+H]+:313.2。 ESI-MS[M+H] + :313.2.
步骤二：D-7-2的合成Step 2: Synthesis of D-7-2
将D-7-1(270mg,0.86mmol)溶于5mL二氯甲烷中，向其中加入HCl/dioxane(5mL,4M)。常温反应1h。旋干溶剂得到化合物D-7-2盐酸盐(280mg,粗品)直接用于下一步反应。D-7-1 (270 mg, 0.86 mmol) was dissolved in 5 mL of dichloromethane, and HCl/dioxane (5 mL, 4M) was added thereto. Reaction at room temperature for 1 hour. The solvent was spin-dried to obtain compound D-7-2 hydrochloride (280 mg, crude product), which was directly used in the next reaction.
步骤三：D-7-3的合成Step 3: Synthesis of D-7-3
将化合物D-7-2盐酸盐(280mg,粗品)和对甲酰基苯甲酸甲酯(170mg,1.0mmol)溶于18mL甲醇中，向其中加入催化量醋酸。常温反应2h后，向其中加入氰基硼氢化钠(110mg,1.7mmol)。常温反应过夜。加水(60mL)淬灭反应，水相用二氯甲烷萃取(60mL*3)。有机相合并后用饱和食盐水(20mL)洗涤，再用无水硫酸钠干燥并浓缩。溶质用柱层析纯化(流动相为石油醚：乙酸乙酯＝1：1)得到化合物D-7-3(160mg,产率50％)。Compound D-7-2 hydrochloride (280 mg, crude product) and methyl p-formylbenzoate (170 mg, 1.0 mmol) were dissolved in 18 mL of methanol, and a catalytic amount of acetic acid was added thereto. After reacting at room temperature for 2 hours, sodium cyanoborohydride (110 mg, 1.7 mmol) was added. Reaction at room temperature overnight. Add water (60mL) to quench the reaction, and extract the aqueous phase with dichloromethane (60mL*3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The solute was purified by column chromatography (the mobile phase was petroleum ether: ethyl acetate = 1:1) to obtain compound D-7-3 (160 mg, yield 50%).
ESI-MS[M+H]+:361.2。ESI-MS[M+H] + :361.2.
步骤四：D-7-4的合成Step 4: Synthesis of D-7-4
将化合物D-7-3(80mg,0.2mmol)溶于甲醇/水(12mL/4mL)中，向其中加入氢氧化锂(30mg,0.6mmol)。60℃反应1h。冷至室温，向反应液中用2N稀盐酸调节pH至4。旋干后得到化合物D-7-4(150mg,粗品)直接用于下一步反应。Compound D-7-3 (80 mg, 0.2 mmol) was dissolved in methanol/water (12 mL/4 mL), and lithium hydroxide (30 mg, 0.6 mmol) was added thereto. React at 60°C for 1 hour. Cool to room temperature, and adjust the pH to 4 with 2N dilute hydrochloric acid. After spin-drying, compound D-7-4 (150 mg, crude product) was obtained and used directly for the next reaction.
步骤五：D-7的合成Step 5: Synthesis of D-7
将化合物D-7-4(150mg,粗品)和D-7-5(86mg,0.28mmol)溶于20mL二氯甲烷中。向其中加入HATU(130mg,0.35mmol)和三乙胺(95mg,0.9mmol)。常温反应过夜。加水(20mL)淬灭反应，水相用DCM萃取(80mL*3)。有机相合并后用饱和食盐水(20mL)洗涤，再用无水硫酸钠干燥并浓缩。粗品用柱层析纯化(流动相为二氯甲烷：甲醇＝20:1)后得化合物D-7(7mg,产率5％)。Compounds D-7-4 (150 mg, crude product) and D-7-5 (86 mg, 0.28 mmol) were dissolved in 20 mL of dichloromethane. HATU (130 mg, 0.35 mmol) and triethylamine (95 mg, 0.9 mmol) were added thereto. Reaction at room temperature overnight. Add water (20mL) to quench the reaction, and extract the aqueous phase with DCM (80mL*3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by column chromatography (the mobile phase was dichloromethane: methanol = 20:1) to obtain compound D-7 (7 mg, yield 5%).
ESI-MS[M+H]+:602.3.ESI-MS[M+H] + :602.3.
1H NMR(400MHz,DMSO-d6)δ10.96(s,1H),9.40(br s,1H),9.10(t,J＝5.9Hz,1H),7.88(d,J＝8.2Hz,2H),7.69(d,J＝7.8Hz,1H),7.54(s,1H),7.47-7.41(m,3H),6.88-6.80(m,2H),6.75(dd,J＝8.9,2.8Hz,1H),5.09(dd,J＝13.3,5.1Hz,1H),4.59(d,J＝5.7Hz,2H),4.37(dd,J＝53.7,17.3Hz,2H),3.57(s,2H),3.04-2.96(m,4H),2.94–2.85(m,1H),2.64-2.56(m,1H),2.49–2.47(m,4H),2.43–2.31(m,1H),2.04–1.97(m,1H). 1 H NMR (400MHz, DMSO-d 6 ) δ10.96 (s, 1H), 9.40 (br s, 1H), 9.10 (t, J = 5.9Hz, 1H), 7.88 (d, J = 8.2Hz, 2H ),7.69(d,J＝7.8Hz,1H),7.54(s,1H),7.47-7.41(m,3H),6.88-6.80(m,2H),6.75(dd,J＝8.9,2.8Hz, 1H),5.09(dd,J＝13.3,5.1Hz,1H),4.59(d,J＝5.7Hz,2H),4.37(dd,J＝53.7,17.3Hz,2H),3.57(s,2H), 3.04-2.96(m,4H),2.94–2.85(m,1H),2.64-2.56(m,1H),2.49–2.47(m,4H),2.43–2.31(m,1H),2.04–1.97(m ,1H).
实施例1.7-A：4-((4-(3-氯-4-(羟甲基)苯基)哌嗪-1-基)甲基)-N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基(D-7-A)
Example 1.7-A: 4-((4-(3-chloro-4-(hydroxymethyl)phenyl)piperazin-1-yl)methyl)-N-((2-(2,6-di Oxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl (D-7-A)
除在步骤二中以D-7-A-2替代D-7-1外，参考D-7的合成方法，得到D-7-A。Except for replacing D-7-1 with D-7-A-2 in step 2, refer to the synthesis method of D-7 to obtain D-7-A.
1H NMR(400MHz,DMSO)δ10.97(s,1H),9.11(t,J＝5.9Hz,1H),7.88(d,J＝8.2Hz,2H),7.69(d,J＝7.8Hz,1H),7.54(s,1H),7.47-7.42(m,3H),7.32(d,J＝9.2Hz,1H),6.92-6.86(m,2H),5.14-5.06(m,2H),4.59(d,J＝5.9Hz,2H),4.48-4.24(m,4H),3.58(s,2H),3.18-3.08(m,4H),2.97–2.83(m,1H),2.64-2.55(m,1H),2.49–2.48(m,4H),2.43-2.30(m,1H),2.08–1.88(m,1H).ESI-MS(M+H)+:616.3。 1 H NMR (400MHz, DMSO) δ10.97 (s, 1H), 9.11 (t, J = 5.9Hz, 1H), 7.88 (d, J = 8.2Hz, 2H), 7.69 (d, J = 7.8Hz, 1H),7.54(s,1H),7.47-7.42(m,3H),7.32(d,J＝9.2Hz,1H),6.92-6.86(m,2H),5.14-5.06(m,2H),4.59 (d,J＝5.9Hz,2H),4.48-4.24(m,4H),3.58(s,2H),3.18-3.08(m,4H),2.97–2.83(m,1H),2.64-2.55(m ,1H),2.49–2.48(m,4H),2.43-2.30(m,1H),2.08–1.88(m,1H).ESI-MS(M+H) + :616.3.
D-7-A-2按照如下方法合成：
D-7-A-2 is synthesized as follows:
(1)化合物D-7-A-1的合成(1) Synthesis of compound D-7-A-1
将化合物2-氯-4-氟苯甲酸甲酯(3.0g,16.0mmol)和N-Boc哌嗪(3.56g,19.1mmol)溶于20mL DMSO中。80℃反应2h后，TLC显示反应完成。加水(60mL)淬灭反应，水相用乙酸乙酯萃取(60mL*3)。有机相合并后用饱和食盐水(10mL*3)洗涤，再用无水硫酸钠干燥并浓缩。粗品用柱层析纯化(流动相为石油醚：乙酸乙酯＝5：1)得到化合物D-7-A-1(4.1g,产率72％)。Compounds 2-chloro-4-fluorobenzoic acid methyl ester (3.0g, 16.0mmol) and N-Boc piperazine (3.56g, 19.1mmol) were dissolved in 20mL DMSO. After reacting at 80°C for 2 hours, TLC showed that the reaction was complete. Water (60 mL) was added to quench the reaction, and the aqueous phase was extracted with ethyl acetate (60 mL*3). The organic phases were combined, washed with saturated brine (10mL*3), dried over anhydrous sodium sulfate and concentrated. The crude product was purified by column chromatography (the mobile phase was petroleum ether: ethyl acetate = 5:1) to obtain compound D-7-A-1 (4.1 g, yield 72%).
(2)化合物D-7-A-2的合成(2) Synthesis of compound D-7-A-2
将化合物D-7-A-1(2.20g,6.21mmol)溶于10mL二氯甲烷中。-20℃下向其中滴加二异丁基氢化铝(12mL,1M,12mmol)。-20℃反应2h后，TLC显示反应完成。加饱和氯化铵溶液(60mL)淬灭反应，水相用二氯甲烷萃取(50mL*3)。有机相合并后用饱和食盐水(10mL*3)洗涤，再用无水硫酸钠干燥并浓缩。粗 品用柱层析纯化(流动相为石油醚：乙酸乙酯＝3：1)得到化合物D-7-A-2(1.4g,产率69％)。Compound D-7-A-1 (2.20 g, 6.21 mmol) was dissolved in 10 mL of dichloromethane. Diisobutylaluminum hydride (12 mL, 1 M, 12 mmol) was added dropwise at -20°C. After reacting at -20°C for 2 hours, TLC showed that the reaction was complete. Add saturated ammonium chloride solution (60mL) to quench the reaction, and extract the aqueous phase with dichloromethane (50mL*3). The organic phases were combined, washed with saturated brine (10mL*3), dried over anhydrous sodium sulfate and concentrated. thick The product was purified by column chromatography (mobile phase was petroleum ether: ethyl acetate = 3:1) to obtain compound D-7-A-2 (1.4g, yield 69%).
实施例1.8 4-((4-(3-氯-4-(2-羟基乙氧基)苯基)哌嗪-1-基)甲基)-N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)苯甲酰胺(D-8)
Example 1.8 4-((4-(3-chloro-4-(2-hydroxyethoxy)phenyl)piperazin-1-yl)methyl)-N-((2-(2,6-di Oxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)benzamide (D-8)
步骤一：D-8-1的合成Step 1: Synthesis of D-8-1
将化合物D-7-3(80mg,0.2mmol)和化合物苄基2-溴乙基醚(75mg,0.3mmol)溶于12mL DMF中，向其中加入碳酸铯(145mg,0.4mmol)。90℃反应2h。加水(60mL)淬灭反应，水相用乙酸乙酯萃取(60mL*3)。有机相合并后用饱和食盐水(10mL*3)洗涤，再用无水硫酸钠干燥并浓缩。粗品用柱层析纯化(流动相为石油醚：乙酸乙酯＝4：1)得到化合物D-8-1(85mg,产率77％)。Compound D-7-3 (80 mg, 0.2 mmol) and compound benzyl 2-bromoethyl ether (75 mg, 0.3 mmol) were dissolved in 12 mL DMF, and cesium carbonate (145 mg, 0.4 mmol) was added thereto. React at 90°C for 2 hours. Water (60 mL) was added to quench the reaction, and the aqueous phase was extracted with ethyl acetate (60 mL*3). The organic phases were combined, washed with saturated brine (10mL*3), dried over anhydrous sodium sulfate and concentrated. The crude product was purified by column chromatography (mobile phase was petroleum ether: ethyl acetate = 4:1) to obtain compound D-8-1 (85 mg, yield 77%).
ESI-MS[M+H]+:495.2。ESI-MS[M+H] + :495.2.
步骤二和步骤三：D-8-2和D-8-3合成操作参考D-7-4和D-7Step 2 and Step 3: For the synthesis operation of D-8-2 and D-8-3, please refer to D-7-4 and D-7
步骤四：D-8的合成Step 4: Synthesis of D-8
将化合物D-8-3(25mg,0.034mmol)溶于10mL DCM中，冰浴下向其中加入BCl3(1mL,1M)溶液。常温反应1h。加甲醇(5mL)淬灭反应后旋干溶剂。粗品经柱层析纯化(流动相为二氯甲烷：甲醇＝20:1)得化合物D-8(15mg,产率68％)。Compound D-8-3 (25 mg, 0.034 mmol) was dissolved in 10 mL DCM, and BCl 3 (1 mL, 1 M) solution was added thereto under ice bath. Reaction at room temperature for 1 hour. Add methanol (5 mL) to quench the reaction and spin the solvent to dryness. The crude product was purified by column chromatography (mobile phase: dichloromethane: methanol = 20:1) to obtain compound D-8 (15 mg, yield 68%).
ESI-MS[M+H]+:646.3。ESI-MS[M+H] + :646.3.
1H NMR(400MHz,DMSO-d6)δ9.11(t,J＝6.0Hz,1H),7.88(d,J＝8.1Hz,2H),7.69(d,J＝7.8Hz,1H),7.54(s,1H),7.48-7.42(m,3H),7.02(d,J＝9.0Hz, 1H),6.97(d,J＝2.8Hz,1H),6.84(dd,J＝9.0,2.8Hz,1H),5.09(dd,J＝13.3,5.1Hz,1H),4.59(d,J＝5.8Hz,2H),4.37(dd,J＝53.8,17.3Hz,2H),3.96(t,J＝5.1Hz,2H),3.69(t,J＝5.1Hz,2H),3.57(s,2H),3.10-3.00(m,4H),2.95–2.85(m,1H),2.64–2.54(m,1H),2.49–2.48(m,4H),2.43–2.30(m,1H),2.04-1.97(m,1H). 1 H NMR (400MHz, DMSO-d 6 ) δ9.11 (t, J = 6.0 Hz, 1H), 7.88 (d, J = 8.1 Hz, 2H), 7.69 (d, J = 7.8 Hz, 1H), 7.54 (s,1H),7.48-7.42(m,3H),7.02(d,J＝9.0Hz, 1H),6.97(d,J＝2.8Hz,1H),6.84(dd,J＝9.0,2.8Hz,1H),5.09(dd,J＝13.3,5.1Hz,1H),4.59(d,J＝5.8 Hz,2H),4.37(dd,J＝53.8,17.3Hz,2H),3.96(t,J＝5.1Hz,2H),3.69(t,J＝5.1Hz,2H),3.57(s,2H), 3.10-3.00(m,4H),2.95–2.85(m,1H),2.64–2.54(m,1H),2.49–2.48(m,4H),2.43–2.30(m,1H),2.04-1.97(m ,1H).
实施例1.9 4-((4-(3-氯-4-甲基苯基)哌嗪-1-基)甲基)-N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-2-羟基苯甲酰胺(D-9)
Example 1.9 4-((4-(3-chloro-4-methylphenyl)piperazin-1-yl)methyl)-N-((2-(2,6-dioxopiperidine-3) -yl)-1-oxoisoindolin-5-yl)methyl)-2-hydroxybenzamide (D-9)
步骤一：化合物D-9-1的合成Step 1: Synthesis of compound D-9-1
将化合物4-甲基水杨酸甲酯(500mg，3.0mmol)溶于15mL四氯化碳中，向其中加入NBS(587mg，3.3mmol)和AIBN(148mg，0.9mmol)。80℃搅拌过夜。加水(40mL)淬灭反应，水相用二氯甲烷萃取(20mL*3)。有机相合并后用饱和食盐水(10mL*3)洗涤，再用无水硫酸钠干燥并旋干得化合物D-9-1(800mg,粗品)，直接用于下一步反应。Compound 4-methylsalicylate methyl ester (500 mg, 3.0 mmol) was dissolved in 15 mL of carbon tetrachloride, and NBS (587 mg, 3.3 mmol) and AIBN (148 mg, 0.9 mmol) were added thereto. Stir overnight at 80°C. Add water (40mL) to quench the reaction, and extract the aqueous phase with dichloromethane (20mL*3). The organic phases were combined, washed with saturated brine (10 mL*3), dried over anhydrous sodium sulfate and spin-dried to obtain compound D-9-1 (800 mg, crude product), which was directly used in the next reaction.
步骤二：化合物D-9-2的合成Step 2: Synthesis of compound D-9-2
将化合物D-9-1(800mg，粗品)和N-Boc哌嗪(558mg,3.0mmol)溶于8mL四氢呋喃中，向其中加入三乙胺(303mg,3.0mmol)。常温反应2h后，TLC显示反应毕。加水(40mL)淬灭反应，水相用乙酸乙酯萃取(30mL*3)。有机相合并后用饱和食盐水(10mL*3)洗涤，再用无水硫酸钠干燥并旋干。粗品经柱层析纯化(流动相为石油醚：乙酸乙酯＝6:1)得化合物D-9-2(544mg,产率52％)。Compound D-9-1 (800 mg, crude product) and N-Boc piperazine (558 mg, 3.0 mmol) were dissolved in 8 mL of tetrahydrofuran, and triethylamine (303 mg, 3.0 mmol) was added thereto. After reacting at room temperature for 2 hours, TLC showed that the reaction was complete. Water (40 mL) was added to quench the reaction, and the aqueous phase was extracted with ethyl acetate (30 mL*3). The organic phases were combined, washed with saturated brine (10 mL*3), dried over anhydrous sodium sulfate, and spun to dryness. The crude product was purified by column chromatography (mobile phase was petroleum ether: ethyl acetate = 6:1) to obtain compound D-9-2 (544 mg, yield 52%).
ESI-MS[M+H]+:351.2。 ESI-MS[M+H] + :351.2.
步骤三：化合物D-9-3的合成Step 3: Synthesis of compound D-9-3
将D-9-2(144mg,0.41mmol)溶于5mL二氯甲烷中，向其中加入HCl/dioxane(5mL,4M)。常温反应1h。旋干溶剂得到化合物D-9-3的盐酸盐(150mg,粗品)直接用于下一步反应。D-9-2 (144 mg, 0.41 mmol) was dissolved in 5 mL of dichloromethane, and HCl/dioxane (5 mL, 4M) was added thereto. Reaction at room temperature for 1 hour. The solvent was spin-dried to obtain the hydrochloride salt of compound D-9-3 (150 mg, crude product), which was directly used in the next reaction.
步骤四：D-9-4的合成Step 4: Synthesis of D-9-4
将D-9-3盐酸盐(150mg,粗品)和2-氯-4-碘-甲基苯(103mg,0.41mmol)溶于6mL二氧六环中，向其中加入叔丁醇钾(92mg,0.82mmol)，Pd2(dba)3(56mg,0.062mmol)和Xphos(39mg,0.082mmol)。氮气保护下100℃反应过夜。加水(40mL)淬灭反应，水相用乙酸乙酯萃取(40mL*3)。合并有机相后，用饱和食盐水(20mL)洗涤。有机相用无水硫酸钠干燥并浓缩。粗品用层析柱纯化(流动相为石油醚：乙酸乙酯＝2：1)得到化合物D-9-4(50mg,产率80％)。Dissolve D-9-3 hydrochloride (150 mg, crude product) and 2-chloro-4-iodo-methylbenzene (103 mg, 0.41 mmol) in 6 mL of dioxane, and add potassium tert-butoxide (92 mg ,0.82mmol), Pd 2 (dba) 3 (56mg, 0.062mmol) and Xphos (39mg, 0.082mmol). The reaction was carried out at 100°C overnight under nitrogen protection. Water (40 mL) was added to quench the reaction, and the aqueous phase was extracted with ethyl acetate (40 mL*3). The organic phases were combined and washed with saturated brine (20 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated. The crude product was purified using a chromatography column (the mobile phase was petroleum ether: ethyl acetate = 2:1) to obtain compound D-9-4 (50 mg, yield 80%).
ESI-MS[M+H]+:375.2。ESI-MS[M+H] + :375.2.
步骤五：D-9-5的合成Step 5: Synthesis of D-9-5
D-9-5的合成操作参考D-7-4。For the synthesis operation of D-9-5, please refer to D-7-4.
步骤六：D-9的合成Step 6: Synthesis of D-9
D-9的合成操作参考D-7。For the synthesis operation of D-9, please refer to D-7.
ESI-MS[M+H]+:616.3。ESI-MS[M+H] + :616.3.
1H NMR(400MHz,DMSO-d6)δ10.97(s,1H),9.39(t,J＝5.9Hz,1H),7.88(d,J＝8.1Hz,1H),7.71(d,J＝7.7Hz,1H),7.56(s,1H),7.48(d,J＝7.4Hz,1H),7.15(d,J＝8.5Hz,1H),6.94-6.86(m,3H),6.82(dd,J＝8.4,2.5Hz,1H),5.10(dd,J＝13.3,5.1Hz,1H),4.63(d,J＝6.1Hz,2H),4.39(dd,J＝54.6,17.5Hz,2H),3.51(s,2H),3.16-3.09(m,4H),2.97–2.85(m,1H),2.68–2.56(m,1H),2.49–2.48(m,4H),2.42–2.31(m,1H),2.21(s,3H),2.07-1.97(m,1H). 1 H NMR (400MHz, DMSO-d 6 ) δ10.97 (s, 1H), 9.39 (t, J = 5.9 Hz, 1H), 7.88 (d, J = 8.1 Hz, 1H), 7.71 (d, J = 7.7Hz,1H),7.56(s,1H),7.48(d,J＝7.4Hz,1H),7.15(d,J＝8.5Hz,1H),6.94-6.86(m,3H),6.82(dd, J＝8.4,2.5Hz,1H),5.10(dd,J＝13.3,5.1Hz,1H),4.63(d,J＝6.1Hz,2H),4.39(dd,J＝54.6,17.5Hz,2H), 3.51(s,2H),3.16-3.09(m,4H),2.97–2.85(m,1H),2.68–2.56(m,1H),2.49–2.48(m,4H),2.42–2.31(m,1H ),2.21(s,3H),2.07-1.97(m,1H).
实施例1.9-A：4-((4-(3-氯-4-甲基苯基)哌嗪-1-基)甲基)-N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-2-(2-羟基乙氧基)苯甲酰胺(D-9-A)
Example 1.9-A: 4-((4-(3-chloro-4-methylphenyl)piperazin-1-yl)methyl)-N-((2-(2,6-dioxopiperine) (Din-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(2-hydroxyethoxy)benzamide (D-9-A)
除采用D-9-A-1替代D-9-4外，参考D-9的制备方法，得到D-9-AIn addition to using D-9-A-1 instead of D-9-4, refer to the preparation method of D-9 to obtain D-9-A
1H NMR(400MHz,DMSO)δ10.97(s,1H),8.94(t,J＝6.0Hz,1H),7.79(d,J＝7.9Hz,1H),7.69(d,J＝7.8Hz,1H),7.57(s,1H),7.49(d,J＝7.9Hz,1H),7.19–7.11(m,2H),7.03(d,J＝8.0Hz,1H),6.92(d,J＝2.4Hz,1H),6.82(dd,J＝8.5,2.5Hz,1H),5.14-5.04(m,2H),4.62(d,J＝6.0Hz,2H),4.37(dd,J＝53.2,17.4Hz,2H),4.19(t,J＝4.7Hz,2H),3.81-3.74(m,2H),3.56(s,2H),3.17-3.06(m,4H),2.96–2.84(m,1H),2.65-2.55(m,1H),2.52-2.51(m,4H),2.44-2.31(m,1H),2.20(s,3H),2.04–1.93(m,1H).ESI-MS(M+H)+:660.3。 1 H NMR (400MHz, DMSO) δ10.97 (s, 1H), 8.94 (t, J = 6.0Hz, 1H), 7.79 (d, J = 7.9Hz, 1H), 7.69 (d, J = 7.8Hz, 1H),7.57(s,1H),7.49(d,J＝7.9Hz,1H),7.19–7.11(m,2H),7.03(d,J＝8.0Hz,1H),6.92(d,J＝2.4 Hz,1H),6.82(dd,J＝8.5,2.5Hz,1H),5.14-5.04(m,2H),4.62(d,J＝6.0Hz,2H),4.37(dd,J＝53.2,17.4Hz ,2H),4.19(t,J＝4.7Hz,2H),3.81-3.74(m,2H),3.56(s,2H),3.17-3.06(m,4H),2.96–2.84(m,1H), 2.65-2.55(m,1H),2.52-2.51(m,4H),2.44-2.31(m,1H),2.20(s,3H),2.04–1.93(m,1H).ESI-MS(M+H ) + :660.3.
D-9-A-1按如下方法制备而成：
D-9-A-1 is prepared as follows:
(1)化合物D-9-A-1的合成(1) Synthesis of compound D-9-A-1
将化合物D-9-4(96mg,0.26mmol)和化合物2-溴乙醇(38mg,0.3mmol)溶于5mL DMF中，向其中加入碳酸钾(55mg,0.4mmol)。80℃反应2h后，TLC显示反应完成。加水(20mL)淬灭反应，水相用乙酸乙酯萃取(20mL*3)。有机相合并后用饱和食盐水(10mL*3)洗涤，再用无水硫酸钠干燥并浓缩。粗品用柱层析纯化(流动相为石油醚：乙酸乙酯＝4：1)得到化合物D-9-A-1(26mg,产率24％)。Compound D-9-4 (96 mg, 0.26 mmol) and compound 2-bromoethanol (38 mg, 0.3 mmol) were dissolved in 5 mL DMF, and potassium carbonate (55 mg, 0.4 mmol) was added thereto. After reacting at 80°C for 2 hours, TLC showed that the reaction was complete. Water (20 mL) was added to quench the reaction, and the aqueous phase was extracted with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (10mL*3), dried over anhydrous sodium sulfate and concentrated. The crude product was purified by column chromatography (the mobile phase was petroleum ether: ethyl acetate = 4:1) to obtain compound D-9-A-1 (26 mg, yield 24%).
实施例1.9-B：4-((4-(3-氯-4-甲基苯基)-2-(羟甲基)哌嗪-1-基)甲基)-N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)苯甲酰胺(D-9-B)
Example 1.9-B: 4-((4-(3-chloro-4-methylphenyl)-2-(hydroxymethyl)piperazin-1-yl)methyl)-N-((2-( 2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)benzamide (D-9-B)
除采用D-9-B-1替代D-9-2外，参考D-9的制备方法，得到D-9-B。 Except using D-9-B-1 instead of D-9-2, refer to the preparation method of D-9 to obtain D-9-B.
1H NMR(400MHz,DMSO)δ10.97(s,1H),9.09(t,J＝6.0Hz,1H),7.87(d,J＝7.9Hz,2H),7.69(d,J＝7.8Hz,1H),7.54(s,1H),7.47-7.43(m,3H),7.15(d,J＝8.5Hz,1H),6.90(s,1H),6.81(dd,J＝8.5,2.1Hz,1H),5.10(dd,J＝13.3,5.1Hz,1H),4.68(s,1H),4.59(d,J＝5.8Hz,2H),4.37(dd,J＝54.5,17.4Hz,2H),4.13(d,J＝13.8Hz,1H),3.80–3.68(m,1H),3.60–3.37(m,3H),3.29–3.25(m,1H),2.96–2.86(m,1H),2.79(t,J＝10.0Hz,2H),2.73–2.54(m,3H),2.42–2.25(m,2H),2.20(s,3H),2.03–1.90(m,1H).ESI-MS(M+H)+:630.3。 1 H NMR (400MHz, DMSO) δ10.97 (s, 1H), 9.09 (t, J = 6.0Hz, 1H), 7.87 (d, J = 7.9Hz, 2H), 7.69 (d, J = 7.8Hz, 1H),7.54(s,1H),7.47-7.43(m,3H),7.15(d,J＝8.5Hz,1H),6.90(s,1H),6.81(dd,J＝8.5,2.1Hz,1H ),5.10(dd,J＝13.3,5.1Hz,1H),4.68(s,1H),4.59(d,J＝5.8Hz,2H),4.37(dd,J＝54.5,17.4Hz,2H),4.13 (d,J＝13.8Hz,1H),3.80–3.68(m,1H),3.60–3.37(m,3H),3.29–3.25(m,1H),2.96–2.86(m,1H),2.79(t ,J＝10.0Hz,2H),2.73–2.54(m,3H),2.42–2.25(m,2H),2.20(s,3H),2.03–1.90(m,1H).ESI-MS(M+H ) + :630.3.
D-9-B-1按如下方法合成：
D-9-B-1 is synthesized as follows:
将化合物1-Boc-3-羟甲基哌嗪(1.05g,5.0mmol)和对甲酰基苯甲酸甲酯(820mg,5.0mmol)溶于20mL甲醇中，向其中加入催化量醋酸。常温反应30min后，向其中加入氰基硼氢化钠(630mg,10.0mmol)。常温反应过夜后，TLC显示反应完成。加水(60mL)淬灭反应，水相用二氯甲烷萃取(60mL*3)。有机相合并后用饱和食盐水(20mL)洗涤，再用无水硫酸钠干燥并浓缩。粗品用柱层析纯化(流动相为石油醚：乙酸乙酯＝1：1)得到化合物D-9-B-1(1.25g,产率69％)。Compound 1-Boc-3-hydroxymethylpiperazine (1.05g, 5.0mmol) and methyl p-formylbenzoate (820mg, 5.0mmol) were dissolved in 20mL methanol, and a catalytic amount of acetic acid was added thereto. After reacting at room temperature for 30 minutes, sodium cyanoborohydride (630 mg, 10.0 mmol) was added. After reacting at room temperature overnight, TLC showed that the reaction was complete. Add water (60mL) to quench the reaction, and extract the aqueous phase with dichloromethane (60mL*3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by column chromatography (the mobile phase was petroleum ether: ethyl acetate = 1:1) to obtain compound D-9-B-1 (1.25 g, yield 69%).
实施例1.9-C：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-2-羟基-4-((甲基(苯乙基)氨基)甲基)苯甲酰胺(D-9-C)
Example 1.9-C: N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-hydroxy-4 -((Methyl(phenylethyl)amino)methyl)benzamide (D-9-C)
除采用D-9-C-1替代D-9-4外，参考D-9的制备方法，得到D-9-CIn addition to using D-9-C-1 instead of D-9-4, refer to the preparation method of D-9 to obtain D-9-C
1H NMR(400MHz,DMSO)δ10.97(s,1H),9.38(t,J＝5.9Hz,1H),7.82(d,J＝8.1Hz,1H),7.69(d,J＝8.1Hz,1H),7.55(s,1H),7.47(d,J＝7.9Hz,1H),7.28-7.24(m,2H),7.20-7.15(m,3H),6.86–6.76(m,2H),5.08(dd,J＝13.3,5.1Hz,1H),4.61(d,J＝5.7Hz,2H),4.37(dd,J＝54.7,17.4Hz,2H),3.51(s,2H),2.94- 2.84(m,1H),2.80–2.73(m,2H),2.65–2.55(m,3H),2.44-2.31(m,1H),2.20(s,3H),2.04-1.94(m,1H).ESI-MS(M+H)+:541.3。 1 H NMR (400MHz, DMSO) δ10.97 (s, 1H), 9.38 (t, J = 5.9Hz, 1H), 7.82 (d, J = 8.1Hz, 1H), 7.69 (d, J = 8.1Hz, 1H),7.55(s,1H),7.47(d,J＝7.9Hz,1H),7.28-7.24(m,2H),7.20-7.15(m,3H),6.86–6.76(m,2H),5.08 (dd,J＝13.3,5.1Hz,1H),4.61(d,J＝5.7Hz,2H),4.37(dd,J＝54.7,17.4Hz,2H),3.51(s,2H),2.94- 2.84(m,1H),2.80–2.73(m,2H),2.65–2.55(m,3H),2.44-2.31(m,1H),2.20(s,3H),2.04-1.94(m,1H). ESI-MS(M+H) + :541.3.
D-9-C-1合成方法如下： The synthesis method of D-9-C-1 is as follows:
将化合物N-甲基-2-苯基乙胺(373mg,2.77mmol)，D-9-1(450mg,1.84mmol)和N,N-二异丙基乙胺(593mg,4.6mmol)溶于8mL乙腈中。80℃反应过夜后，TLC显示反应完成。加水(25mL)淬灭反应，水相用乙酸乙酯萃取(25mL*3)。有机相合并后用饱和食盐水(10mL*3)洗涤，再用无水硫酸钠干燥并浓缩。粗品用柱层析纯化(流动相为石油醚：乙酸乙酯＝10：1)得到化合物D-9-C-1(300mg,产率54％)。Compounds N-methyl-2-phenylethylamine (373mg, 2.77mmol), D-9-1 (450mg, 1.84mmol) and N,N-diisopropylethylamine (593mg, 4.6mmol) were dissolved in in 8 mL acetonitrile. After reacting at 80°C overnight, TLC showed that the reaction was complete. Water (25 mL) was added to quench the reaction, and the aqueous phase was extracted with ethyl acetate (25 mL*3). The organic phases were combined, washed with saturated brine (10mL*3), dried over anhydrous sodium sulfate and concentrated. The crude product was purified by column chromatography (the mobile phase was petroleum ether: ethyl acetate = 10:1) to obtain compound D-9-C-1 (300 mg, yield 54%).
实施例1.10：N-(4-(((3,4-二氯苯基乙基)氨基)甲基)苄基)-2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-甲酰胺(D-10)的合成
Example 1.10: N-(4-(((3,4-dichlorophenylethyl)amino)methyl)benzyl)-2-(2,6-dioxopiperidin-3-yl)- Synthesis of 1-oxoisoindoline-5-carboxamide (D-10)
步骤一：化合物D-10-1的合成Step 1: Synthesis of compound D-10-1
2-(3,4-二氯苯基)乙醇(500mg,2.63mmol)溶于12mL DCM中，冰浴下向其中分批加入Dess-Martin试剂(1.67g,3.95mmol)。室温下反应2h后，TLC显示反应完成。将反应液过滤后，滤液加aq.NaHCO3(30mL)。用二氯甲烷萃取(30mL*3)。有机相合并后，用无水硫酸钠干燥，过滤，旋干溶剂。粗品用柱层析纯化(洗脱剂为石油醚：乙酸乙酯＝100:1–15:1)，得到目标化合物D-10-1(390mg)。2-(3,4-Dichlorophenyl)ethanol (500 mg, 2.63 mmol) was dissolved in 12 mL DCM, and Dess-Martin reagent (1.67 g, 3.95 mmol) was added in batches under ice bath. After reacting for 2 hours at room temperature, TLC showed that the reaction was complete. After filtering the reaction solution, aq.NaHCO 3 (30 mL) was added to the filtrate. Extract with dichloromethane (30mL*3). After the organic phases were combined, they were dried over anhydrous sodium sulfate, filtered, and the solvent was spin-dried. The crude product was purified by column chromatography (eluent was petroleum ether: ethyl acetate = 100:1–15:1) to obtain the target compound D-10-1 (390 mg).
步骤二：化合物D-10的合成Step 2: Synthesis of Compound D-10
将化合物D-10-1(390mg,2.06mmol)，D-1-5(830mg,2.0mmol)和醋酸(100mg)溶于20mL甲醇中。室温反应0.5h后，向其中加入氰基硼氢化钠(260mg,4.13mmol)。继续反应过夜后，TLC显示反应完成。向其中加饱和食盐水(30mL)后，用二氯甲烷萃取(35mL*3)。有机相合并后，用无水硫酸钠干燥， 过滤，旋干溶剂。粗品用柱层析纯化(洗脱剂为二氯甲烷：甲醇＝20:1)，得目标化合物D-10(160mg)。Compound D-10-1 (390 mg, 2.06 mmol), D-1-5 (830 mg, 2.0 mmol) and acetic acid (100 mg) were dissolved in 20 mL methanol. After reacting at room temperature for 0.5 h, sodium cyanoborohydride (260 mg, 4.13 mmol) was added. After continuing the reaction overnight, TLC showed that the reaction was complete. After adding saturated brine (30 mL), the mixture was extracted with dichloromethane (35 mL*3). After the organic phases were combined, they were dried over anhydrous sodium sulfate. Filter and spin dry the solvent. The crude product was purified by column chromatography (eluent: dichloromethane: methanol = 20:1) to obtain the target compound D-10 (160 mg).
ESI-MS[M+H]+:579.2。ESI-MS[M+H] + :579.2.
1H NMR(400MHz,DMSO-d6)δ11.00(s,1H),9.20(t,J＝6.0Hz,1H),8.09(s,1H),8.01(d,J＝8.0Hz,1H),7.82(d,J＝7.9Hz,1H),7.54–7.47(m,2H),7.28(s,4H),7.21(dd,J＝8.2,2.0Hz,1H),5.14(dd,J＝13.2,5.1Hz,1H),4.56–4.35(m,4H),3.73(s,2H),2.97–2.88(m,1H),2.81–2.69(n,4H),2.65–2.56(m,1H),2.46–2.37(m,1H),2.08–1.92(m,1H). 1 H NMR (400MHz, DMSO-d 6 ) δ11.00 (s, 1H), 9.20 (t, J = 6.0Hz, 1H), 8.09 (s, 1H), 8.01 (d, J = 8.0Hz, 1H) ,7.82(d,J＝7.9Hz,1H),7.54–7.47(m,2H),7.28(s,4H),7.21(dd,J＝8.2,2.0Hz,1H),5.14(dd,J＝13.2 ,5.1Hz,1H),4.56–4.35(m,4H),3.73(s,2H),2.97–2.88(m,1H),2.81–2.69(n,4H),2.65–2.56(m,1H), 2.46–2.37(m,1H),2.08–1.92(m,1H).
实施例1.10-A：2-(2,6-二氧代哌啶-3-基)-N-(4-(((2-(萘-1-基)乙基)氨基)甲基)苄基)-1-氧代异吲哚啉-5-甲酰胺(D-10-A)
Example 1.10-A: 2-(2,6-dioxopiperidin-3-yl)-N-(4-(((2-(naphth-1-yl)ethyl)amino)methyl)benzyl base)-1-oxoisoindoline-5-carboxamide (D-10-A)
除采用1-萘乙醇替代2-(3,4-二氯苯基)乙醇外，参考D-10的制备方法，得到D-10-A。Except using 1-naphthyl ethanol instead of 2-(3,4-dichlorophenyl)ethanol, refer to the preparation method of D-10 to obtain D-10-A.
1H NMR(400MHz,DMSO)δ11.01(s,1H),9.23(t,J＝5.9Hz,1H),8.12–7.97(m,3H),7.94–7.87(m,1H),7.82(d,J＝7.9Hz,1H),7.78-7.75(m,1H),7.54–7.25(m,8H),5.14(dd,J＝13.3,5.1Hz,1H),4.59–4.34(m,4H),3.83(s,2H),3.25-3.20(m,2H),2.92–2.83(m,3H),2.69–2.55(m,1H),2.46–2.34(m,1H),2.07–1.90(m,1H).ESI-MS(M+H)+:560.3. 1 H NMR (400MHz, DMSO) δ11.01 (s, 1H), 9.23 (t, J = 5.9Hz, 1H), 8.12–7.97 (m, 3H), 7.94–7.87 (m, 1H), 7.82 (d ,J＝7.9Hz,1H),7.78-7.75(m,1H),7.54–7.25(m,8H),5.14(dd,J＝13.3,5.1Hz,1H),4.59–4.34(m,4H), 3.83(s,2H),3.25-3.20(m,2H),2.92–2.83(m,3H),2.69–2.55(m,1H),2.46–2.34(m,1H),2.07–1.90(m,1H ).ESI-MS(M+H) + :560.3.
实施例1.11：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-4-(((2-羟基-2-苯乙基)(甲基)氨基)甲基)苯甲酰胺(D-11)
Example 1.11: N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-4-(((2- Hydroxy-2-phenylethyl)(methyl)amino)methyl)benzamide (D-11)
步骤一：D-11-1的合成Step 1: Synthesis of D-11-1
将化合物4-溴甲基苯甲酸甲酯(500mg,2.2mmol)溶于15mL甲醇中，向其中加入甲胺醇溶液(30％，6mL)。室温反应1h后，TLC显示反应完成。向其中加水(30mL)后，用二氯甲烷萃取(35mL*3)。有机相合并后，用无水硫酸钠干燥，过滤，旋干溶剂。粗品用柱层析纯化(洗脱剂为二氯甲烷：甲醇＝20：1)，得化合物D-11-1(230mg,产率57％)。Compound 4-bromomethylbenzoic acid methyl ester (500 mg, 2.2 mmol) was dissolved in 15 mL of methanol, and methylamine alcohol solution (30%, 6 mL) was added thereto. After reacting at room temperature for 1 hour, TLC showed that the reaction was complete. After adding water (30 mL), the mixture was extracted with dichloromethane (35 mL*3). After the organic phases were combined, they were dried over anhydrous sodium sulfate, filtered, and the solvent was spin-dried. The crude product was purified by column chromatography (eluent: dichloromethane: methanol = 20:1) to obtain compound D-11-1 (230 mg, yield 57%).
步骤二：D-11-2的合成Step 2: Synthesis of D-11-2
将化合物D-11-1(230mg,1.3mmol)和DIPEA(660mg,5.1mmol)溶于15mL乙腈中，向其中加入2-溴苯乙酮(310mg,1.5mmol)。60℃反应2h后，TLC显示反应完成。向其中加水(20mL)后，用乙酸乙酯萃取(30mL*3)。有机相合并后，用无水硫酸钠干燥，过滤，旋干溶剂。粗品用柱层析纯化(洗脱剂为石油醚：乙酸乙酯＝4：1)，得化合物D-11-2(250mg,产率65％)。Compound D-11-1 (230 mg, 1.3 mmol) and DIPEA (660 mg, 5.1 mmol) were dissolved in 15 mL of acetonitrile, and 2-bromoacetophenone (310 mg, 1.5 mmol) was added thereto. After reacting at 60°C for 2 hours, TLC showed that the reaction was complete. After adding water (20 mL), the mixture was extracted with ethyl acetate (30 mL*3). After the organic phases were combined, they were dried over anhydrous sodium sulfate, filtered, and the solvent was spin-dried. The crude product was purified by column chromatography (eluent was petroleum ether: ethyl acetate = 4:1) to obtain compound D-11-2 (250 mg, yield 65%).
步骤三：D-11-3的合成Step 3: Synthesis of D-11-3
将化合物D-11-2(250mg,0.8mmol)溶于15mL甲醇中，冰浴下向其中加入硼氢化钠(48mg,1.3mmol)。0℃反应2h后，TLC显示反应完成。向其中加水(20mL)后，用二氯甲烷萃取(30mL*3)。有机相合并后，用无水硫酸钠干燥，过滤，旋干溶剂。粗品用柱层析纯化(洗脱剂为石油醚：乙酸乙酯＝2：1)，得化合物D-11-3(200mg,产率79％)。 Compound D-11-2 (250 mg, 0.8 mmol) was dissolved in 15 mL of methanol, and sodium borohydride (48 mg, 1.3 mmol) was added thereto under ice bath. After reacting at 0°C for 2 hours, TLC showed that the reaction was complete. After adding water (20 mL), the mixture was extracted with dichloromethane (30 mL*3). After the organic phases were combined, they were dried over anhydrous sodium sulfate, filtered, and the solvent was spin-dried. The crude product was purified by column chromatography (eluent was petroleum ether: ethyl acetate = 2:1) to obtain compound D-11-3 (200 mg, yield 79%).
D-11-4和D-11合成操作分别参考D-7-4和D-7的制备方法，最终得到D-11
The synthesis operations of D-11-4 and D-11 refer to the preparation methods of D-7-4 and D-7 respectively, and finally D-11 is obtained.
1H NMR(400MHz,DMSO)δ10.97(s,1H),9.09(t,J＝5.9Hz,1H),7.82(d,J＝8.2Hz,2H),7.69(d,J＝7.8Hz,1H),7.54(s,1H),7.46(d,J＝7.9Hz,1H),7.36–7.27(m,6H),7.26–7.19(m,1H),5.10(dd,J＝13.3,5.1Hz,1H),4.74(dd,J＝7.6,5.1Hz,1H),4.58(d,J＝5.9Hz,2H),4.37(dd,J＝54.6,17.4Hz,2H),3.60(s,2H),2.96–2.84(m,1H),2.65–2.53(m,2H),2.49–2.44(m,1H),2.43-2.32(m,2H),2.22(s,3H),2.05-1.95(m,1H).ESI-MS(M+H)+:541.3。 1 H NMR (400MHz, DMSO) δ10.97 (s, 1H), 9.09 (t, J = 5.9Hz, 1H), 7.82 (d, J = 8.2Hz, 2H), 7.69 (d, J = 7.8Hz, 1H),7.54(s,1H),7.46(d,J＝7.9Hz,1H),7.36–7.27(m,6H),7.26–7.19(m,1H),5.10(dd,J＝13.3,5.1Hz ,1H),4.74(dd,J＝7.6,5.1Hz,1H),4.58(d,J＝5.9Hz,2H),4.37(dd,J＝54.6,17.4Hz,2H),3.60(s,2H) ,2.96–2.84(m,1H),2.65–2.53(m,2H),2.49–2.44(m,1H),2.43-2.32(m,2H),2.22(s,3H),2.05-1.95(m, 1H).ESI-MS(M+H) + :541.3.
实施例1.12：2-(2,6-二氧代哌啶-3-基)-N-(4-((3-(2-羟乙基)-1-苯乙基脲基)甲基)苄基)-1-氧代异吲哚啉-5-甲酰胺(D-12)
Example 1.12: 2-(2,6-dioxopiperidin-3-yl)-N-(4-((3-(2-hydroxyethyl)-1-phenylethylureido)methyl) Benzyl)-1-oxoisoindoline-5-carboxamide (D-12)
步骤一：D-12-1的合成Step 1: Synthesis of D-12-1
将化合物2-苄氧基-1-乙胺(30mg,0.2mmol)溶于5mL DMF中，向其中加入N,N-羰基二咪唑(34mg，0.2mL)。室温反应2h后，向其中加入D-1-6(50mg,0.1mmol)。0℃反应2h后，TLC显示反应完成。向其中加水(30mL)后，用乙酸乙酯萃取(35mL*3)。有机相合并后，用无水硫酸钠干燥，过滤，旋干溶剂。粗品用柱层析纯化(洗脱剂为二氯甲烷：甲醇＝20：1)，得化合物D-12-1(35mg,产率70％)。Compound 2-benzyloxy-1-ethylamine (30 mg, 0.2 mmol) was dissolved in 5 mL DMF, and N, N-carbonyldiimidazole (34 mg, 0.2 mL) was added thereto. After reacting at room temperature for 2 hours, D-1-6 (50 mg, 0.1 mmol) was added thereto. After reacting at 0°C for 2 hours, TLC showed that the reaction was complete. After adding water (30 mL), the mixture was extracted with ethyl acetate (35 mL*3). After the organic phases were combined, they were dried over anhydrous sodium sulfate, filtered, and the solvent was spin-dried. The crude product was purified by column chromatography (eluent: dichloromethane: methanol = 20:1) to obtain compound D-12-1 (35 mg, yield 70%).
步骤二中合成操作参考D-8的制备，得到D-12.
The synthesis operation in step 2 refers to the preparation of D-8, and D-12 is obtained.
1H NMR(400MHz,DMSO)δ11.00(s,1H),9.19(t,J＝5.9Hz,1H),8.08(s,1H),8.00(d,J＝8.0Hz,1H),7.81(d,J＝7.9Hz,1H),7.30-7.24(m,4H),7.19-7.15(m,5H),6.38(t,J＝5.5Hz,1H),5.14(dd,J＝13.2,5.1Hz,1H),4.61(t,J＝5.4Hz,1H),4.56–4.32(m,6H),3.39(dd,J＝11.8,6.1Hz,2H),3.29-3.24(m,2H),3.13(q,J＝6.1Hz,2H),2.97–2.86(m,1H),2.75–2.68(m,2H),2.66-2.55(m,1H),2.48–2.35(m,1H),2.06–1.97(m,1H).ESI-MS(M+H)+:598.3。 1 H NMR (400MHz, DMSO) δ11.00(s,1H),9.19(t,J＝5.9Hz,1H),8.08(s,1H),8.00(d,J＝8.0Hz,1H),7.81( d,J＝7.9Hz,1H),7.30-7.24(m,4H),7.19-7.15(m,5H),6.38(t,J＝5.5Hz,1H),5.14(dd,J＝13.2,5.1Hz ,1H),4.61(t,J＝5.4Hz,1H),4.56–4.32(m,6H),3.39(dd,J＝11.8,6.1Hz,2H),3.29-3.24(m,2H),3.13( q,J＝6.1Hz,2H),2.97–2.86(m,1H),2.75–2.68(m,2H),2.66-2.55(m,1H),2.48–2.35(m,1H),2.06–1.97( m,1H).ESI-MS(M+H) + :598.3.
实施例1.13：2-(1-(5-溴-3-(羟甲基)吡啶-2-基)哌啶-4-基)-N-(2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)乙酰胺(D-13)
Example 1.13: 2-(1-(5-bromo-3-(hydroxymethyl)pyridin-2-yl)piperidin-4-yl)-N-(2-(2,6-dioxopiperidine) -3-yl)-1-oxoisoindolin-5-yl)acetamide (D-13)
步骤一：化合物D-13-1的合成Step 1: Synthesis of compound D-13-1
将化合物5-溴-2-氯-3-甲醛吡啶(250mg,1.14mmol)溶于5mL DMF中，向其中加入哌啶乙酸甲酯(264mg，1.37mL)。80℃反应2h后，TLC显示反应完成。向其中加水(30mL)后，用乙酸乙酯萃取(35mL*3)。有机相合并后用饱和食盐水洗(10mL*3)。有机相用无水硫酸钠干燥，过滤，旋干溶剂。粗品用柱层析纯化(洗脱剂为石油醚：乙酸乙酯＝5：1)，得化合物D-13-1(197mg,产率50％)。Compound 5-bromo-2-chloro-3-carboxaldehyde pyridine (250 mg, 1.14 mmol) was dissolved in 5 mL DMF, and piperidine acetate methyl ester (264 mg, 1.37 mL) was added thereto. After reacting at 80°C for 2 hours, TLC showed that the reaction was complete. After adding water (30 mL), the mixture was extracted with ethyl acetate (35 mL*3). Combine the organic phases and wash with saturated brine (10mL*3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the solvent was spun dry. The crude product was purified by column chromatography (eluent was petroleum ether: ethyl acetate = 5:1) to obtain compound D-13-1 (197 mg, yield 50%).
步骤二：化合物D-13-2的合成Step 2: Synthesis of compound D-13-2
将化合物D-13-1(197mg,0.58mmol)溶于6mL甲醇中，冰浴下向其中加入硼氢化钠(33mg,0.87mmol)。0℃反应2h后，TLC显示反应完成。向 其中加水(20mL)后，用乙酸乙酯萃取(30mL*3)。有机相合并后，用无水硫酸钠干燥，过滤，旋干溶剂。粗品用柱层析纯化(洗脱剂为石油醚：乙酸乙酯＝5：1)，得化合物D-13-2(190mg,产率97％)。Compound D-13-1 (197 mg, 0.58 mmol) was dissolved in 6 mL of methanol, and sodium borohydride (33 mg, 0.87 mmol) was added thereto under ice bath. After reacting at 0°C for 2 hours, TLC showed that the reaction was complete. Towards After adding water (20mL), extract with ethyl acetate (30mL*3). After the organic phases were combined, they were dried over anhydrous sodium sulfate, filtered, and the solvent was spin-dried. The crude product was purified by column chromatography (eluent was petroleum ether: ethyl acetate = 5:1) to obtain compound D-13-2 (190 mg, yield 97%).
步骤三和步骤四中D-13-3和D-13合成操作参考D-7-4和D-7的制备，得到D-13。
The synthesis operations of D-13-3 and D-13 in Steps 3 and 4 refer to the preparation of D-7-4 and D-7 to obtain D-13.
1H NMR(400MHz,DMSO)δ10.97(s,1H),10.26(s,1H),8.21(d,J＝2.5Hz,1H),8.00(s,1H),7.86(d,J＝2.4Hz,1H),7.65(d,J＝8.3Hz,1H),7.59(d,J＝9.6Hz,1H),5.08(dd,J＝13.1,5.0Hz,1H),4.47–4.23(m,4H),3.34–3.26(m,2H),2.96–2.83(m,1H),2.74(t,J＝11.8Hz,2H),2.64-2.54(m,1H),2.43–2.28(m,3H),2.04–1.93(m,2H),1.81-1.69(m,2H),1.43-1.30(m,2H).ESI-MS(M+H)+:570.3。 1 H NMR (400MHz, DMSO) δ10.97 (s, 1H), 10.26 (s, 1H), 8.21 (d, J = 2.5Hz, 1H), 8.00 (s, 1H), 7.86 (d, J = 2.4 Hz,1H),7.65(d,J＝8.3Hz,1H),7.59(d,J＝9.6Hz,1H),5.08(dd,J＝13.1,5.0Hz,1H),4.47–4.23(m,4H ),3.34–3.26(m,2H),2.96–2.83(m,1H),2.74(t,J＝11.8Hz,2H),2.64-2.54(m,1H),2.43–2.28(m,3H), 2.04–1.93(m,2H),1.81-1.69(m,2H),1.43-1.30(m,2H).ESI-MS(M+H) + :570.3.
实施例1.13-A：N-(2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)-2-(1-(3-(羟甲基)-5-(喹啉-8-基)吡啶-2-基)哌啶-4-基)乙酰胺(D-13-A)Example 1.13-A: N-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)-2-(1-(3-( Hydroxymethyl)-5-(quinolin-8-yl)pyridin-2-yl)piperidin-4-yl)acetamide (D-13-A)
除采用D-13-A-2替代D-13-2外，参考D-13方法制备得到D-13-A。
Except using D-13-A-2 instead of D-13-2, D-13-A was prepared by referring to the method of D-13.
1H NMR(400MHz,DMSO)δ10.97(s,1H),10.28(s,1H),8.91(dd,J＝4.1,1.7Hz,1H),8.51–8.38(m,2H),8.07(d,J＝2.2Hz,1H),8.03-7.97(m,2H),7.82–7.75(m,1H),7.73-7.64(m,2H),7.64–7.53(m,2H),5.28(t,J＝5.4Hz,1H),5.08(dd,J＝13.2,5.1Hz,1H),4.55(d,J＝5.1Hz,2H),4.37(dd,J＝56.0,17.3Hz,2H),3.54-3.44(m,2H),3.01–2.81(m,3H),2.66-2.55(m,1H),2.44–2.26(m,3H),2.06-1.93(m,2H),1.87-1.77(m,2H),1.52-1.40(m,2H).ESI-MS(M+H)+:619.3。 1 H NMR (400MHz, DMSO) δ10.97 (s, 1H), 10.28 (s, 1H), 8.91 (dd, J = 4.1, 1.7Hz, 1H), 8.51–8.38 (m, 2H), 8.07 (d ,J＝2.2Hz,1H),8.03-7.97(m,2H),7.82–7.75(m,1H),7.73-7.64(m,2H),7.64–7.53(m,2H),5.28(t,J ＝5.4Hz,1H),5.08(dd,J＝13.2,5.1Hz,1H),4.55(d,J＝5.1Hz,2H),4.37(dd,J＝56.0,17.3Hz,2H),3.54-3.44 (m,2H),3.01–2.81(m,3H),2.66-2.55(m,1H),2.44–2.26(m,3H),2.06-1.93(m,2H),1.87-1.77(m,2H) ,1.52-1.40(m,2H).ESI-MS(M+H) + :619.3.
D-13-A对应的中间体D-13-A-2制备方法如下：
The preparation method of intermediate D-13-A-2 corresponding to D-13-A is as follows:
(1)化合物D-13-A-1的合成(1) Synthesis of compound D-13-A-1
将喹啉-8-硼酸(870mg,5.0mmol)，D-13-1(1.7g,0.90mmol)和碳酸钾(2.07g,15.0mmol)溶于二氧六环/水(20mL/5mL)中，向其中加入Pd(dppf)Cl2(366mg,0.5mmol)。氮气置换三次，在90℃下反应3h后，TLC显示反应完成。将反应液过滤，滤液用乙酸乙酯(60mL)稀释后，有机相用饱和食盐水洗(20ml*3)后溶液旋干。粗品经柱层析(石油醚:乙酸乙酯＝5:1)分离纯化得化合物D-13-A-1(300mg，产率88％)。Dissolve quinoline-8-boronic acid (870mg, 5.0mmol), D-13-1 (1.7g, 0.90mmol) and potassium carbonate (2.07g, 15.0mmol) in dioxane/water (20mL/5mL) , to which Pd(dppf)Cl 2 (366 mg, 0.5 mmol) was added. Nitrogen was replaced three times, and after reacting at 90°C for 3 hours, TLC showed that the reaction was complete. Filter the reaction solution, dilute the filtrate with ethyl acetate (60 mL), wash the organic phase with saturated brine (20 ml*3), and spin the solution to dryness. The crude product was separated and purified by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound D-13-A-1 (300 mg, yield 88%).
(2)化合物D-13-A-2的合成(2) Synthesis of compound D-13-A-2
将化合物D-13-A-1(780mg,2.0mmol)溶于40mL甲醇中，冰浴下向其中加入硼氢化钠(152mg,4.0mmol)。0℃反应2h后，TLC显示反应完成。向其中加水(60mL)后，用乙酸乙酯萃取(60mL*3)。有机相合并后，用无水硫酸钠干燥，过滤，旋干溶剂。粗品用柱层析纯化(洗脱剂为石油醚：乙酸乙酯＝1：1)，得化合物D-13-A-2(640mg,产率82％)。Compound D-13-A-1 (780 mg, 2.0 mmol) was dissolved in 40 mL of methanol, and sodium borohydride (152 mg, 4.0 mmol) was added thereto under ice bath. After reacting at 0°C for 2 hours, TLC showed that the reaction was complete. After adding water (60 mL), the mixture was extracted with ethyl acetate (60 mL*3). After the organic phases were combined, they were dried over anhydrous sodium sulfate, filtered, and the solvent was spin-dried. The crude product was purified by column chromatography (eluent was petroleum ether: ethyl acetate = 1:1) to obtain compound D-13-A-2 (640 mg, yield 82%).
实施例1.13-B：N-(2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)-2-(1-(3-(羟甲基)-5-(异喹啉-1-基)吡啶-2-基)哌啶-4-基)乙酰胺(D-13-B)Example 1.13-B: N-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)-2-(1-(3-( Hydroxymethyl)-5-(isoquinolin-1-yl)pyridin-2-yl)piperidin-4-yl)acetamide (D-13-B)
除采用D-13-B-2替代D-13-1外，参考D-13方法制备得到D-13-B。
Except using D-13-B-2 instead of D-13-1, D-13-B was prepared by referring to the D-13 method.
1H NMR(400MHz,DMSO)δ10.97(s,1H),10.29(s,1H),8.59(d,J＝5.6Hz,1H),8.46(d,J＝2.3Hz,1H),8.13–8.00(m,4H),7.86-7.78(m,2H),7.71–7.59(m,3H),5.37(t,J＝5.4Hz,1H),5.08(dd,J＝13.3,5.1Hz,1H),4.56(d,J＝5.0Hz,2H),4.35(dd,J＝56.0,17.4Hz,2H),3.62-3.51(m,2H),2.96–2.82(m,3H),2.66–2.56(m,1H),2.44-2.32(m,3H),2.07-1.96(m,2H),1.87-1.77(m,2H),1.53-1.35(m,2H).ESI-MS(M+H)+:619.4 1 H NMR (400MHz, DMSO) δ10.97(s,1H),10.29(s,1H),8.59(d,J＝5.6Hz,1H),8.46(d,J＝2.3Hz,1H),8.13– 8.00(m,4H),7.86-7.78(m,2H),7.71–7.59(m,3H),5.37(t,J＝5.4Hz,1H),5.08(dd,J＝13.3,5.1Hz,1H) ,4.56(d,J＝5.0Hz,2H),4.35(dd,J＝56.0,17.4Hz,2H),3.62-3.51(m,2H),2.96–2.82(m,3H),2.66–2.56(m ,1H),2.44-2.32(m,3H),2.07-1.96(m,2H),1.87-1.77(m,2H),1.53-1.35(m,2H).ESI-MS(M+H) + : 619.4
D-13-B-2制备方法如下：
The preparation method of D-13-B-2 is as follows:
(1)化合物D-13-B-1的合成(1) Synthesis of compound D-13-B-1
将频哪醇硼酸酯(2.35g,9.26mmol)，D-13-1(2.1g,6.18mmol)和醋酸钾(1.51g,15.45mmol)溶于20mL二氧六环中，向其中加入Pd(dppf)Cl2(454mg,0.62mmol)。氮气置换三次，在90℃下反应过夜后，TLC显示反应完成。将反应液过滤，滤液用乙酸乙酯(60mL)稀释后，有机相用饱和食盐水洗(20ml*3)后溶液旋干。粗品经柱层析(石油醚:乙酸乙酯＝5:1)分离纯化得化合物D-13-B-1(1.86g，产率77％)。Dissolve pinacol borate (2.35g, 9.26mmol), D-13-1 (2.1g, 6.18mmol) and potassium acetate (1.51g, 15.45mmol) in 20mL of dioxane, and add Pd to it (dppf)Cl 2 (454 mg, 0.62 mmol). After nitrogen replacement three times and reaction at 90°C overnight, TLC showed that the reaction was complete. Filter the reaction solution, dilute the filtrate with ethyl acetate (60 mL), wash the organic phase with saturated brine (20 ml*3), and spin the solution to dryness. The crude product was separated and purified by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound D-13-B-1 (1.86 g, yield 77%).
(2)化合物D-13-B-2的合成(2) Synthesis of compound D-13-B-2
将1-溴异喹啉(290mg,1.4mmol)，D-13-B-1(504mg,1.3mmol)和碳酸钾(449mg,15.0mmol)溶于二氧六环/水(4mL/1mL)中，向其中加入Pd(dppf)Cl2(95mg,0.62mmol)。氮气置换三次，在80℃下反应过夜后，TLC显示反应完成。将反应液过滤，滤液用乙酸乙酯(60mL)稀释后，有机相用饱和食盐水洗(20ml*3)后溶液旋干。粗品经柱层析(石油醚:乙酸乙酯＝3:1)分离纯化得化合物D-13-B-2(408mg，产率81％)。Dissolve 1-bromoisoquinoline (290mg, 1.4mmol), D-13-B-1 (504mg, 1.3mmol) and potassium carbonate (449mg, 15.0mmol) in dioxane/water (4mL/1mL) , to which Pd(dppf)Cl 2 (95 mg, 0.62 mmol) was added. After nitrogen replacement three times and reaction at 80°C overnight, TLC showed that the reaction was complete. Filter the reaction solution, dilute the filtrate with ethyl acetate (60 mL), wash the organic phase with saturated brine (20 ml*3), and spin the solution to dryness. The crude product was separated and purified by column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain compound D-13-B-2 (408 mg, yield 81%).
实施例1.14：N-(2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)-2-(1-(3-(羟甲基)吡啶-2-基)哌啶-4-基)乙酰胺(D-14)
Example 1.14: N-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)-2-(1-(3-(hydroxymethyl) base)pyridin-2-yl)piperidin-4-yl)acetamide (D-14)
D-14是由D-13通过还原制备而成，具体方法如下：D-14 is prepared from D-13 by reduction. The specific method is as follows:
将化合物D-13(60mg,0.10mmol)和钯碳(5％,10mg)溶于5mL甲醇中。在氢气氛围下，常温反应2h后，TLC显示反应完成。将反应液过滤，滤饼用甲醇淋洗后，滤液旋干。粗品用柱层析纯化(洗脱剂为二氯甲烷：甲醇＝15：1)，得化合物D-14(8mg,产率16％)。 Compound D-13 (60 mg, 0.10 mmol) and palladium on carbon (5%, 10 mg) were dissolved in 5 mL of methanol. After reacting for 2 hours at room temperature under a hydrogen atmosphere, TLC showed that the reaction was completed. The reaction solution was filtered, the filter cake was washed with methanol, and the filtrate was spun to dryness. The crude product was purified by column chromatography (eluent: dichloromethane: methanol = 15:1) to obtain compound D-14 (8 mg, yield 16%).
1H NMR(400MHz,DMSO)δ10.97(s,1H),10.27(s,1H),8.12(dd,J＝4.8,1.9Hz,1H),8.01(s,1H),7.75(dd,J＝7.4,1.8Hz,1H),7.66(d,J＝8.3Hz,1H),7.60(dd,J＝8.3,1.6Hz,1H),6.98(dd,J＝7.4,4.8Hz,1H),5.25(s,1H),5.08(dd,J＝13.3,5.1Hz,1H),4.50–4.22(m,4H),3.34–3.22(m,2H),2.97–2.81(m,1H),2.73(t,J＝11.6Hz,2H),2.64-2.54(m,1H),2.45–2.29(m,3H),2.05–1.92(m,2H),1.81-1.72(m,2H),1.45-1.32(m,2H).ESI-MS(M+H)+:492.3。 1 H NMR (400MHz, DMSO) δ10.97(s,1H),10.27(s,1H),8.12(dd,J＝4.8,1.9Hz,1H),8.01(s,1H),7.75(dd,J ＝7.4,1.8Hz,1H),7.66(d,J＝8.3Hz,1H),7.60(dd,J＝8.3,1.6Hz,1H),6.98(dd,J＝7.4,4.8Hz,1H),5.25 (s,1H),5.08(dd,J＝13.3,5.1Hz,1H),4.50–4.22(m,4H),3.34–3.22(m,2H),2.97–2.81(m,1H),2.73(t ,J＝11.6Hz,2H),2.64-2.54(m,1H),2.45–2.29(m,3H),2.05–1.92(m,2H),1.81-1.72(m,2H),1.45-1.32(m ,2H).ESI-MS(M+H) + :492.3.
实施例1.15：2-(1-(3-(氨基甲基)-5-(喹啉-8-基)吡啶-2-基)哌啶-4-基)-N-(2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)乙酰胺(D-15)
Example 1.15: 2-(1-(3-(aminomethyl)-5-(quinolin-8-yl)pyridin-2-yl)piperidin-4-yl)-N-(2-(2, 6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)acetamide (D-15)
步骤一：D-15-1的制备Step 1: Preparation of D-15-1
将化合物D-13-A-2(640mg,1.63mmol)和1,8-二氮杂双环[5.4.0]十一碳-7-烯(380mg,2.5mmol)溶于20mL甲苯中，向其中加入叠氮磷酸二苯酯(550mg,2.0mmol)。常温反应3h后，TLC显示反应完成。向其中加水(30mL)后，用乙酸乙酯萃取(30mL*3)。有机相合并后，用无水硫酸钠干燥，过滤，旋干溶剂得粗品化合物D-15-1(680mg,粗品)直接用于下一步反应。Compound D-13-A-2 (640 mg, 1.63 mmol) and 1,8-diazabicyclo[5.4.0]undec-7-ene (380 mg, 2.5 mmol) were dissolved in 20 mL of toluene. Add diphenyl phosphate azide (550 mg, 2.0 mmol). After reacting at room temperature for 3 hours, TLC showed that the reaction was complete. After adding water (30 mL), the mixture was extracted with ethyl acetate (30 mL*3). After the organic phases were combined, they were dried over anhydrous sodium sulfate, filtered, and the solvent was spin-dried to obtain crude compound D-15-1 (680 mg, crude product), which was directly used in the next reaction.
步骤二：D-15-2的制备Step 2: Preparation of D-15-2
将化合物D-15-1(680mg,1.63mmol)，二碳酸二叔丁酯(540mg,2.5mmol)和钯碳(5％，50mg)溶于10mL甲醇中。在氢气氛围下，常温反应5h后，TLC显示反应完成。将反应液过滤，滤饼用甲醇淋洗后，滤液旋干。粗品用柱层析纯化(洗脱剂为石油醚：乙酸乙酯＝1：1)，得化合物D-15-2(350mg,产率44％)。 Compound D-15-1 (680 mg, 1.63 mmol), di-tert-butyl dicarbonate (540 mg, 2.5 mmol) and palladium on carbon (5%, 50 mg) were dissolved in 10 mL of methanol. After reacting for 5 hours at room temperature under a hydrogen atmosphere, TLC showed that the reaction was completed. The reaction solution was filtered, the filter cake was washed with methanol, and the filtrate was spun to dryness. The crude product was purified by column chromatography (eluent was petroleum ether: ethyl acetate = 1:1) to obtain compound D-15-2 (350 mg, yield 44%).
D-15-3和D-15-4合成操作分别参考D-7-4和D-7的制备方法，然后以得到的D-15-4为原料，按照如下方法制备D-15：For the synthesis operations of D-15-3 and D-15-4, refer to the preparation methods of D-7-4 and D-7 respectively, and then use the obtained D-15-4 as raw material to prepare D-15 according to the following method:
将化合物D-15-4(108mg,0.15mmol)溶于5mL乙酸乙酯中，向其中加入HCl/dioxane(4M,5mL)。常温反应2h后，TLC显示反应完成。将反应液旋干。粗品用柱层析纯化(洗脱剂为二氯甲烷：甲醇＝10：1)，得化合物D-15(43mg,产率47％)。
Compound D-15-4 (108 mg, 0.15 mmol) was dissolved in 5 mL of ethyl acetate, and HCl/dioxane (4 M, 5 mL) was added thereto. After reacting at room temperature for 2 hours, TLC showed that the reaction was complete. Spin the reaction solution dry. The crude product was purified by column chromatography (eluent: dichloromethane: methanol = 10:1) to obtain compound D-15 (43 mg, yield 47%).
1H NMR(400MHz,DMSO)δ10.33(s,1H),8.91(dd,J＝4.1,1.7Hz,1H),8.50(d,J＝2.2Hz,1H),8.44(dd,J＝8.3,1.7Hz,1H),8.12(d,J＝2.0Hz,1H),8.05–7.96(m,2H),7.81(dd,J＝7.1,1.2Hz,1H),7.74–7.64(m,2H),7.64–7.54(m,2H),5.07(dd,J＝13.3,5.1Hz,1H),4.36(dd,J＝56.0,17.4Hz,2H),3.94(s,2H),3.54-3.44(m,2H),2.97-2.80(m,3H),2.65–2.56(m,1H),2.46–2.27(m,3H),2.07–1.92(m,2H),1.87-1.77(m,2H),1.55-1.41(m,2H).ESI-MS(M+H)+:618.4。 1 H NMR (400MHz, DMSO) δ10.33 (s, 1H), 8.91 (dd, J = 4.1, 1.7Hz, 1H), 8.50 (d, J = 2.2Hz, 1H), 8.44 (dd, J = 8.3 ,1.7Hz,1H),8.12(d,J＝2.0Hz,1H),8.05–7.96(m,2H),7.81(dd,J＝7.1,1.2Hz,1H),7.74–7.64(m,2H) ,7.64–7.54(m,2H),5.07(dd,J＝13.3,5.1Hz,1H),4.36(dd,J＝56.0,17.4Hz,2H),3.94(s,2H),3.54-3.44(m ,2H),2.97-2.80(m,3H),2.65–2.56(m,1H),2.46–2.27(m,3H),2.07–1.92(m,2H),1.87-1.77(m,2H),1.55 -1.41(m,2H).ESI-MS(M+H) + :618.4.
实施例1.16：N-(2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)-2-(1-(3-(2-羟基乙氧基)-5-(喹啉-8-基)吡啶-2-基)哌啶-4-基)乙酰胺(D-16)
Example 1.16: N-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)-2-(1-(3-(2- Hydroxyethoxy)-5-(quinolin-8-yl)pyridin-2-yl)piperidin-4-yl)acetamide (D-16)
步骤一：D-16-1的制备Step 1: Preparation of D-16-1
将化合物5-溴-2-氟-3-羟基吡啶(500mg,2.6mmol)和化合物苄基2-溴乙基醚(840mg,0.3mmol)溶于15mL DMF中，向其中加入碳酸铯(1.7g,5.2mmol)。120℃反应2h后，TLC显示反应完成。加水(60mL)淬灭反应，水相用乙酸乙酯萃取(60mL*3)。有机相合并后用饱和食盐水(10mL*3)洗涤，再用无水硫酸钠干燥并浓缩。粗品用柱层析纯化(流动相为石油醚：乙酸乙酯＝10：1)得到化合物D-16-1(800mg,产率94％)。Compound 5-bromo-2-fluoro-3-hydroxypyridine (500 mg, 2.6 mmol) and compound benzyl 2-bromoethyl ether (840 mg, 0.3 mmol) were dissolved in 15 mL DMF, and cesium carbonate (1.7 g was added) ,5.2mmol). After reacting at 120°C for 2 hours, TLC showed that the reaction was complete. Water (60 mL) was added to quench the reaction, and the aqueous phase was extracted with ethyl acetate (60 mL*3). The organic phases were combined, washed with saturated brine (10mL*3), dried over anhydrous sodium sulfate and concentrated. The crude product was purified by column chromatography (the mobile phase was petroleum ether: ethyl acetate = 10:1) to obtain compound D-16-1 (800 mg, yield 94%).
D-16-2，D-16-3，D-16-4和D-16-5合成操作分别参考D-13-1,D-13-A-1,D-7-4和D-7，得到D-16-5；然后以D-16-5为原料，参考D-8的制备方法合成D-16.
The synthesis operations of D-16-2, D-16-3, D-16-4 and D-16-5 refer to D-13-1, D-13-A-1, D-7-4 and D-7 respectively. , obtain D-16-5; then use D-16-5 as raw material and synthesize D-16 by referring to the preparation method of D-8.
1H NMR(400MHz,DMSO)δ10.97(s,1H),10.27(s,1H),8.92(dd,J＝4.1,1.8Hz,1H),8.43(dd,J＝8.3,1.7Hz,1H),8.09(d,J＝1.8Hz,1H),8.02-7.97(m,2H),7.82(dd,J＝7.1,1.3Hz,1H),7.74–7.47(m,5H),5.08(dd,J＝13.3,5.1Hz,1H),4.85(br s,1H),4.36(dd,J＝56.2,17.4Hz,2H),4.17-4.08(m,2H),4.06(t,J＝4.9Hz,2H),3.81-3.72(m,2H),2.97–2.85(m,1H),2.84-2.74(m,2H),2.65-2.56(m,1H),2.44–2.31(m,3H),2.07-1.95(m,2H),1.83-1.72(m,2H),1.50-1.36(m,2H).ESI-MS(M+H)+:649.6。 1 H NMR (400MHz, DMSO) δ10.97(s,1H),10.27(s,1H),8.92(dd,J＝4.1,1.8Hz,1H),8.43(dd,J＝8.3,1.7Hz,1H ),8.09(d,J＝1.8Hz,1H),8.02-7.97(m,2H),7.82(dd,J＝7.1,1.3Hz,1H),7.74–7.47(m,5H),5.08(dd, J＝13.3,5.1Hz,1H),4.85(br s,1H),4.36(dd,J＝56.2,17.4Hz,2H),4.17-4.08(m,2H),4.06(t,J＝4.9Hz, 2H),3.81-3.72(m,2H),2.97–2.85(m,1H),2.84-2.74(m,2H),2.65-2.56(m,1H),2.44–2.31(m,3H),2.07- 1.95(m,2H),1.83-1.72(m,2H),1.50-1.36(m,2H).ESI-MS(M+H) + :649.6.
实施例1.17：2-(1-(3-(2-氨基乙氧基)-5-(喹啉-8-基)吡啶-2-基)哌啶-4-基)-N-(2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)乙酰胺(D-17)
Example 1.17: 2-(1-(3-(2-aminoethoxy)-5-(quinolin-8-yl)pyridin-2-yl)piperidin-4-yl)-N-(2- (2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)acetamide (D-17)
步骤一：D-17-1的合成Step 1: Synthesis of D-17-1
将化合物D-16-3(2g,3.9mmol)溶于40mL二氯甲烷中，冰浴下向其中加入BCl3(11.7mL,1M)溶液。常温反应1h，TLC显示反应完成。加甲醇(10mL)淬灭反应后旋干溶剂。粗品经柱层析纯化(流动相为二氯甲烷：甲醇＝10:1)得化合物D-17-1(1.5g,产率91％)。Compound D-16-3 (2g, 3.9mmol) was dissolved in 40mL dichloromethane, and BCl 3 (11.7mL, 1M) solution was added thereto under ice bath. The reaction was carried out at room temperature for 1 hour, and TLC showed that the reaction was completed. Add methanol (10 mL) to quench the reaction and spin the solvent to dryness. The crude product was purified by column chromatography (the mobile phase was dichloromethane: methanol = 10:1) to obtain compound D-17-1 (1.5 g, yield 91%).
步骤二：D-17-2的合成Step 2: Synthesis of D-17-2
将化合物D-17-1(1.4g,3.3mmol)和三乙胺(670mg,6.6mmol)溶于20mL二氯甲烷中，冰浴下向其中加入甲磺酰氯(570mg,4.9mmol)。常温反应3h，TLC显示反应完成。加水(10mL)淬灭反应后，水相用二氯甲烷萃取(40mL*3)。有机相合并后用无水硫酸钠干燥，过滤，旋干溶剂。粗品经柱层析纯化(流动相为石油醚：乙酸乙酯＝1:1)得化合物D-17-2(1.15g,产率69％)。Compound D-17-1 (1.4g, 3.3mmol) and triethylamine (670mg, 6.6mmol) were dissolved in 20mL of methylene chloride, and methanesulfonyl chloride (570mg, 4.9mmol) was added thereto under ice bath. The reaction was carried out at room temperature for 3 hours, and TLC showed that the reaction was completed. After adding water (10 mL) to quench the reaction, the aqueous phase was extracted with dichloromethane (40 mL*3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the solvent was spun dry. The crude product was purified by column chromatography (mobile phase was petroleum ether: ethyl acetate = 1:1) to obtain compound D-17-2 (1.15g, yield 69%).
步骤三：D-17-3的合成Step 3: Synthesis of D-17-3
将化合物D-17-2(1.15g,2.3mmol)溶于20mL DMF中，向其中加入叠氮化钠(200mg,3mmol)。60℃反应过夜后，TLC显示反应完成。加水(30mL)淬灭反应后，水相用乙酸乙酯萃取(30mL*3)。有机相合并后用饱和 食盐水洗(10mL*3)后有机相无水硫酸钠干燥，过滤，旋干溶剂。粗品经柱层析纯化(流动相为石油醚：乙酸乙酯＝1:1)得化合物D-17-3(1.03g,产率100％)。Compound D-17-2 (1.15g, 2.3mmol) was dissolved in 20mL DMF, and sodium azide (200mg, 3mmol) was added thereto. After reacting at 60°C overnight, TLC showed that the reaction was complete. After adding water (30 mL) to quench the reaction, the aqueous phase was extracted with ethyl acetate (30 mL*3). The organic phases were combined and saturated with After washing with brine (10mL*3), the organic phase was dried over anhydrous sodium sulfate, filtered, and the solvent was spin-dried. The crude product was purified by column chromatography (mobile phase was petroleum ether: ethyl acetate = 1:1) to obtain compound D-17-3 (1.03g, yield 100%).
D-17-4合成操作参考D-15-2制备方法，D-17-5和D-17-6合成操作分别参考D-7-3和D-7的制备方法得到中间体D-17-6；然后以D-17-6为原料，参考D-15对应步骤五得到D-17。
For the synthesis operations of D-17-4, refer to the preparation method of D-15-2. For the synthesis operations of D-17-5 and D-17-6, refer to the preparation methods of D-7-3 and D-7 respectively to obtain intermediate D-17- 6; Then use D-17-6 as raw material and refer to step 5 corresponding to D-15 to obtain D-17.
1H NMR(400MHz,DMSO)δ10.32(s,1H),8.91(d,J＝2.6Hz,1H),8.43(d,J＝7.5Hz,1H),8.11(s,1H),8.06–7.94(m,2H),7.81(d,J＝6.9Hz,1H),7.73–7.64(m,2H),7.63–7.55(m,3H),5.07(dd,J＝13.2,5.1Hz,1H),4.36(dd,J＝56.4,17.2Hz,2H),4.14–3.98(m,4H),3.13-2.97(m,2H),2.93–2.73(m,3H),2.66-1.55(m,1H),2.45–2.27(m,3H),2.08–1.93(m,2H),1.84-1.73(m,2H),1.50-1.33(m,2H).ESI-MS(M+H)+:648.4。 1 H NMR (400MHz, DMSO) δ10.32(s,1H),8.91(d,J＝2.6Hz,1H),8.43(d,J＝7.5Hz,1H),8.11(s,1H),8.06– 7.94(m,2H),7.81(d,J＝6.9Hz,1H),7.73–7.64(m,2H),7.63–7.55(m,3H),5.07(dd,J＝13.2,5.1Hz,1H) ,4.36(dd,J＝56.4,17.2Hz,2H),4.14–3.98(m,4H),3.13-2.97(m,2H),2.93–2.73(m,3H),2.66-1.55(m,1H) ,2.45–2.27(m,3H),2.08–1.93(m,2H),1.84-1.73(m,2H),1.50-1.33(m,2H).ESI-MS(M+H) + :648.4.
实施例2.中间体合成Example 2. Intermediate synthesis
实施例2.1：N-((10S,13S)-10-(4-(二丙氨基)丁基)-1,1,1-三氟-14-甲基-6,9,12-三氧代-3-氧杂-5,8,11-三氮杂十五烷-13-基)-6-(2-(甲磺酰基)嘧啶-5-基)己-5-炔酰胺(L-1)的合成
Example 2.1: N-((10S,13S)-10-(4-(dipropylamino)butyl)-1,1,1-trifluoro-14-methyl-6,9,12-trioxo -3-oxa-5,8,11-triazapentadecan-13-yl)-6-(2-(methanesulfonyl)pyrimidin-5-yl)hex-5-ynamide (L-1 )Synthesis
步骤一：化合物L-1-2的合成Step 1: Synthesis of compound L-1-2
将化合物L-1-1(10.00g,27.15mmol)加入至2,2,2-三氟乙醇(60mL)中，40℃搅拌溶解，然后加入HCl/DMF(3.4mL,0.4M)于反应液中，40℃搅拌2.0h。向反应液中加入乙酸乙酯(200mL)，依次用2％的碳酸氢钠溶液(200mL)、饱和食盐水溶液(200mL)洗涤，分离有机相；有机相用无水硫酸钠干燥，过滤，浓缩得目标化合物L-1-2(10.51g)。Add compound L-1-1 (10.00g, 27.15mmol) to 2,2,2-trifluoroethanol (60mL), stir and dissolve at 40°C, then add HCl/DMF (3.4mL, 0.4M) to the reaction solution Medium, stir at 40°C for 2.0h. Ethyl acetate (200 mL) was added to the reaction solution, washed with 2% sodium bicarbonate solution (200 mL) and saturated saline solution (200 mL) in sequence, and the organic phase was separated; the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain Target compound L-1-2 (10.51g).
1H NMR(400MHz,DMSO-d6)δ8.87(t,J＝6.8Hz,1H),7.92(d,J＝7.5Hz,2H),7.74(d,J＝7.5Hz,2H),7.62(t,J＝6.3Hz,1H),7.45(t,J＝7.4Hz,2H),7.36(td,J＝7.4,1.2Hz,2H),4.70(d,J＝6.8Hz,2H),4.33(d,J＝6.9Hz,2H),4.29-4.22(m,1H),4.03(q,J＝9.4Hz,2H),3.68(t,J＝3.1Hz,2H). 1 H NMR (400MHz, DMSO-d 6 ) δ8.87 (t, J = 6.8 Hz, 1H), 7.92 (d, J = 7.5 Hz, 2H), 7.74 (d, J = 7.5 Hz, 2H), 7.62 (t,J＝6.3Hz,1H),7.45(t,J＝7.4Hz,2H),7.36(td,J＝7.4,1.2Hz,2H),4.70(d,J＝6.8Hz,2H),4.33 (d,J＝6.9Hz,2H),4.29-4.22(m,1H),4.03(q,J＝9.4Hz,2H),3.68(t,J＝3.1Hz,2H).
步骤二：化合物L-1-3的合成Step 2: Synthesis of compound L-1-3
将化合物L-1-2(5.7g，13.9mmol)溶于DMF(35mL)和二乙胺(1.7mL)的溶液中。25℃下搅拌1h。35℃减压浓缩30min。得目标化合物L-1-3的DMF溶液，直接用于下一步反应(转化率按照100％计算)。Compound L-1-2 (5.7 g, 13.9 mmol) was dissolved in a solution of DMF (35 mL) and diethylamine (1.7 mL). Stir at 25°C for 1 hour. Concentrate under reduced pressure at 35°C for 30 minutes. A DMF solution of target compound L-1-3 was obtained, which was directly used in the next reaction (conversion rate was calculated as 100%).
步骤三：化合物L-1-5的合成Step 3: Synthesis of compound L-1-5
向步骤二得到的L-1-3的DMF溶液中，加入L-1-4(7.0g，12.7mmol)和DMF(35mL)。将反应液冷却至-15℃，加入DMTMM(4.2g，15.2mmol)，-10℃ 反应搅拌2h。将反应液倒入DCM(200mL)中，有机相依次用2％NaHCO3水溶液(100mL*3)、水(100mL)和饱和NaCl水溶液(50mL)洗涤。有机相无水硫酸钠干燥，滤除不溶物，浓缩得粗品，硅胶柱层析(洗脱剂：0-7％MeOH/DCM)纯化，得目标化合物L-1-5(7.1g)。To the DMF solution of L-1-3 obtained in step 2, add L-1-4 (7.0 g, 12.7 mmol) and DMF (35 mL). Cool the reaction solution to -15°C, add DMTMM (4.2g, 15.2mmol), -10°C The reaction was stirred for 2 h. Pour the reaction solution into DCM (200 mL), and wash the organic phase with 2% NaHCO 3 aqueous solution (100 mL*3), water (100 mL) and saturated NaCl aqueous solution (50 mL). The organic phase was dried over anhydrous sodium sulfate, insoluble matter was filtered off, concentrated to obtain a crude product, and purified by silica gel column chromatography (eluent: 0-7% MeOH/DCM) to obtain the target compound L-1-5 (7.1g).
ESI-MS[M+H]+＝720.5.ESI-MS[M+H] + =720.5.
1H NMR(400MHz，DMSO-d6)δ8.78(t,J＝8.0Hz,1H),8.28(t,J＝8.0Hz,1H),8.02(d,J＝4.0Hz,1H),7.90-7.88(m,2H),7.72(t,J＝8.0Hz,2H),7.46-7.39(m,3H),7.34-7.30(m,2H),4.71-4.62(m,2H),4.30-4.24(m,4H),4.03-3.96(m,2H),3.90-3.86(m,1H),3.80-3.66(m,2H),2.35-2.15(m,6H),1.99-1.94(m,1H),1.66-1.63(m,1H),1.56-1.53(m,1H),1.36-1.26(m,8H),0.89-0.77(m,12H). 1 H NMR (400MHz, DMSO-d 6 ) δ8.78(t,J＝8.0Hz,1H),8.28(t,J＝8.0Hz,1H),8.02(d,J＝4.0Hz,1H),7.90 -7.88(m,2H),7.72(t,J＝8.0Hz,2H),7.46-7.39(m,3H),7.34-7.30(m,2H),4.71-4.62(m,2H),4.30-4.24 (m,4H),4.03-3.96(m,2H),3.90-3.86(m,1H),3.80-3.66(m,2H),2.35-2.15(m,6H),1.99-1.94(m,1H) ,1.66-1.63(m,1H),1.56-1.53(m,1H),1.36-1.26(m,8H),0.89-0.77(m,12H).
步骤四：化合物L-1-6的合成Step 4: Synthesis of compound L-1-6
将化合物L-1-5(1.7g，2.4mmol)溶于DMF(10mL)中，加入二乙胺(0.5mL)，25℃下反应1h。向反应液加入四氢呋喃(15mL)，减压浓缩，此操作重复三次，得目标化合物L-1-6的DMF溶液，直接用于下一步反应(转化率按照100％计算)。Compound L-1-5 (1.7g, 2.4mmol) was dissolved in DMF (10mL), diethylamine (0.5mL) was added, and the reaction was carried out at 25°C for 1 hour. Tetrahydrofuran (15 mL) was added to the reaction solution and concentrated under reduced pressure. This operation was repeated three times to obtain a DMF solution of target compound L-1-6, which was directly used in the next reaction (conversion rate was calculated as 100%).
ESI-MS[M+H]+＝498.6.ESI-MS[M+H] + =498.6.
步骤五：化合物L-1的合成Step 5: Synthesis of Compound L-1
25℃下，向步骤四得到的L-1-6的DMF溶液中依次加入L-1-7(772mg，2.9mmol)和DMTMM(849mg，2.9mmol)，搅拌反应1h。反应液经prep-HPLC纯化、冻干，得目标化合物L-1(1.3g)。At 25°C, add L-1-7 (772 mg, 2.9 mmol) and DMTMM (849 mg, 2.9 mmol) in sequence to the DMF solution of L-1-6 obtained in step 4, and stir for 1 hour. The reaction solution was purified by prep-HPLC and freeze-dried to obtain the target compound L-1 (1.3g).
ESI-MS[M+H]+＝748.4.ESI-MS[M+H] + =748.4.
1H NMR(400MHz,DMSO-d6)δ9.15(s,2H),8.78(t,J＝6.8Hz,1H),8.24(t,J＝5.8Hz,1H),8.05(d,J＝7.3Hz,1H),7.95(d,J＝8.6Hz,1H),4.75-4.66(m,2H),4.24-4.19(m,2H),4.03(q,J＝9.4Hz,2H),3.78-3.76(m,2H),3.44(s,3H),2.58(t,J＝7.1Hz,2H),2.46-2.29(m,8H),2.03-1.95(m,1H),1.89-1.81(m,2H),1.73-1.66(m,1H),1.62-1.56(m,1H),1.41-1.36(m,6H),1.31-1.25(m,2H),0.89-0.83(m,12H). 1 H NMR (400MHz, DMSO-d 6 ) δ9.15 (s, 2H), 8.78 (t, J = 6.8Hz, 1H), 8.24 (t, J = 5.8Hz, 1H), 8.05 (d, J = 7.3Hz,1H),7.95(d,J＝8.6Hz,1H),4.75-4.66(m,2H),4.24-4.19(m,2H),4.03(q,J＝9.4Hz,2H),3.78- 3.76(m,2H),3.44(s,3H),2.58(t,J＝7.1Hz,2H),2.46-2.29(m,8H),2.03-1.95(m,1H),1.89-1.81(m, 2H),1.73-1.66(m,1H),1.62-1.56(m,1H),1.41-1.36(m,6H),1.31-1.25(m,2H),0.89-0.83(m,12H).
实施例2.2：N6,N6-二丙基-N2-((6-(2-(甲磺酰基)嘧啶-5-基)己-5-炔酰基)-L-缬氨酸)-L-赖氨酸(L-2)的合成
Example 2.2: N 6 ,N 6 -dipropyl-N 2 -((6-(2-(methanesulfonyl)pyrimidin-5-yl)hex-5-ynyl)-L-valine)- Synthesis of L-lysine (L-2)
步骤一：step one:
将化合物L-2-1(5.0g，12mmol)溶在二氯甲烷(100mL)中，向反应液中加入正丙醛(4.2g，72.3mmol)，室温下搅拌反应10min，再向反应液中加入三乙酰氧基硼氢化钠(12.8g，60.25mmol)，反应在室温下搅拌反应1h；向反应液中加入氯化铵饱和水溶液搅拌1h，减压浓缩，过滤，滤液用C18柱反向分离纯化(乙腈：H2O(含0.05％的甲酸)＝5％-55％)，得目标化合物L-2-2(4.57g)。Dissolve compound L-2-1 (5.0g, 12mmol) in dichloromethane (100mL), add n-propionaldehyde (4.2g, 72.3mmol) to the reaction solution, stir for 10 minutes at room temperature, and then add Add sodium triacetoxyborohydride (12.8g, 60.25mmol), stir at room temperature for 1 hour; add saturated aqueous ammonium chloride solution to the reaction solution, stir for 1 hour, concentrate under reduced pressure, filter, and use a C18 column to reversely separate the filtrate. Purification (acetonitrile: H 2 O (containing 0.05% formic acid) = 5%-55%) gave the target compound L-2-2 (4.57g).
ESI-MS[M+H]+＝464.0；ESI-MS[M+H] + =464.0;
1H NMR(400MHz,DMSO-d6)δ7.81(d,J＝7.3Hz,1H),7.38–7.28(m,5H),5.04(d,J＝1.7Hz,2H),4.12–4.02(m,1H),3.94–3.82(m,1H),2.65–2.52(m,6H),2.06–1.94(m,1H),1.76–1.64(m,1H),1.64–1.53(m,1H),1.52–1.40(m,6H),1.34–1.18(m,2H),0.92–0.80(m,12H)。 1 H NMR (400MHz, DMSO-d6) δ7.81(d,J＝7.3Hz,1H),7.38–7.28(m,5H),5.04(d,J＝1.7Hz,2H),4.12–4.02(m ,1H),3.94–3.82(m,1H),2.65–2.52(m,6H),2.06–1.94(m,1H),1.76–1.64(m,1H),1.64–1.53(m,1H),1.52 –1.40(m,6H),1.34–1.18(m,2H),0.92–0.80(m,12H).
步骤二：Step two:
室温下，将化合物L-2-2(2.0g，4.32mmol)溶于甲醇(80mL)中，然后向反应液中加入Pd/C(0.16g)，氢气下室温下搅拌反应12h。反应液过滤，滤液减压浓缩，得目标化合物L-2-3(1.2g)。Compound L-2-2 (2.0 g, 4.32 mmol) was dissolved in methanol (80 mL) at room temperature, then Pd/C (0.16 g) was added to the reaction solution, and the reaction was stirred for 12 h at room temperature under hydrogen gas. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain the target compound L-2-3 (1.2g).
步骤三：Step three:
将化合物L-1-7(100mg,0.373mmol)，溶于N,N-二甲基甲酰胺(1mL),然后加入HATU(142mg,0.373mmol)，N,N-二异丙基乙胺(120mg,0.93mmol)，该体系搅拌30min，然后加入化合物L-2-3(122mg，0.371mmol)，反应液室温搅拌1h。反应液直接经C18柱反相纯化(乙腈：H2O(含0.05％的甲酸)＝5％-55％)，得目标化合物L-2(50mg)。Compound L-1-7 (100 mg, 0.373 mmol) was dissolved in N, N-dimethylformamide (1 mL), and then HATU (142 mg, 0.373 mmol), N, N-diisopropylethylamine ( 120 mg, 0.93 mmol), the system was stirred for 30 min, then compound L-2-3 (122 mg, 0.371 mmol) was added, and the reaction solution was stirred at room temperature for 1 h. The reaction solution was directly purified by C18 column reverse phase (acetonitrile: H 2 O (containing 0.05% formic acid) = 5%-55%) to obtain the target compound L-2 (50 mg).
ESI-MS[M+H]+＝580.0. ESI-MS[M+H] + =580.0.
1H NMR(400MHz,DMSO-d6)δ8.24(s,2H),7.98–7.93(m,2H),4.24–4.16(m,1H),4.10(d,J＝5.2Hz,1H),3.41(s,3H),2.79–2.64(m,6H),2.55(t,J＝7.1Hz,2H),2.45–2.26(m,2H),2.06–1.91(m,1H),1.89–1.78(m,2H),1.76–1.66(m,1H),1.64–1.57(m,1H),1.57–1.42(m,6H),1.37–1.24(m,2H),0.93–0.78(m,12H)。 1 H NMR (400MHz, DMSO-d 6 ) δ8.24 (s, 2H), 7.98–7.93 (m, 2H), 4.24–4.16 (m, 1H), 4.10 (d, J = 5.2Hz, 1H), 3.41(s,3H),2.79–2.64(m,6H),2.55(t,J＝7.1Hz,2H),2.45–2.26(m,2H),2.06–1.91(m,1H),1.89–1.78( m,2H),1.76–1.66(m,1H),1.64–1.57(m,1H),1.57–1.42(m,6H),1.37–1.24(m,2H),0.93–0.78(m,12H).
实施例2.3：N-(4-(((2-((2-氨基乙酰胺基)甲氧基)乙基)(苯乙基)氨基)甲基)苄基)-2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-酰胺(L-3)的合成
Example 2.3: N-(4-(((2-((2-aminoacetamido)methoxy)ethyl)(phenylethyl)amino)methyl)benzyl)-2-(2,6 -Synthesis of dioxopiperidin-3-yl)-1-oxoisoindoline-5-amide (L-3)
步骤一：step one:
将化合物D-1(60mg,0.11mmol)，化合物L-1-1(122mg,0.33mmol)加入反应瓶中，再加入DMF(0.2mL)，氩气保护下加入HCl/DMF(0.1mL,0.66M)，反应液在室温下搅拌6h；LCMS监测反应；反应液经制备高效液相色谱纯化(乙腈：水(含0.1％甲酸)＝5％-90％)，得目标化合物L-3-1(33mg)。Add compound D-1 (60 mg, 0.11 mmol) and compound L-1-1 (122 mg, 0.33 mmol) into the reaction bottle, then add DMF (0.2 mL), and add HCl/DMF (0.1 mL, 0.66) under argon protection. M), the reaction solution was stirred at room temperature for 6 hours; LCMS monitored the reaction; the reaction solution was purified by preparative high-performance liquid chromatography (acetonitrile: water (containing 0.1% formic acid) = 5%-90%) to obtain the target compound L-3-1 (33mg).
ESI-MS[M+H]+＝863.4.ESI-MS[M+H] + =863.4.
步骤二：Step two:
将化合物L-3-1(33mg，0.038mmol)加入反应瓶中，然后加入DMF(1mL,含5％二乙胺),室温搅拌1h。反应液直接减压浓缩，得目标化合物L-3(24mg，粗品)。Add compound L-3-1 (33 mg, 0.038 mmol) into the reaction bottle, then add DMF (1 mL, containing 5% diethylamine), and stir at room temperature for 1 h. The reaction solution was directly concentrated under reduced pressure to obtain the target compound L-3 (24 mg, crude product).
LCMS(ESI)[M+H]+＝641.3.LCMS(ESI)[M+H] + =641.3.
实施例3.药物连接体偶联物的合成Example 3. Synthesis of drug linker conjugates
实施例3.1：2-(2,6-二氧代哌啶-3-基)-N-(4-((12S,15S)-12-(4-(二丙基氨基)丁基)-15-异丙基-22-(2-(甲基磺酰基)嘧啶-5-基)-2-(2-(萘-1-基)乙基)-3,8,11,14,17-五 氧代-5-氧杂-2,7,10,13,16-五氮杂二十二碳-21-炔-1-基)苄基)-1-氧代异吲哚啉-5-甲酰胺(DL-1)的合成
Example 3.1: 2-(2,6-dioxopiperidin-3-yl)-N-(4-((12S,15S)-12-(4-(dipropylamino)butyl)-15 -Isopropyl-22-(2-(methylsulfonyl)pyrimidin-5-yl)-2-(2-(naphthyl-1-yl)ethyl)-3,8,11,14,17-penta Oxo-5-oxa-2,7,10,13,16-pentaazabis-21-yn-1-yl)benzyl)-1-oxoisoindoline-5-methyl Synthesis of Amide (DL-1)
步骤一：化合物DL-1的合成Step 1: Synthesis of compound DL-1
在反应瓶中加入化合物D-4(50mg,0.081mmol)，HCl/DMF溶液(300μL,0.243mmol)，搅拌下加入化合物L-1的盐酸盐(127.06mg,0.16mmol)，氮气保护下在室温反应3h。反应液经制备高效液相色谱(乙腈：0.05％甲酸的水溶液＝5％-50％)纯化，得目标化合物的甲酸盐(13.9mg)。Add compound D-4 (50 mg, 0.081 mmol) and HCl/DMF solution (300 μL, 0.243 mmol) to the reaction bottle. Add compound L-1 hydrochloride (127.06 mg, 0.16 mmol) under stirring, and add it under nitrogen protection. React at room temperature for 3 hours. The reaction solution was purified by preparative high performance liquid chromatography (acetonitrile: 0.05% formic acid aqueous solution = 5%-50%) to obtain the formate salt of the target compound (13.9 mg).
ESI-MS[M+H]+＝1266.76.ESI-MS[M+H] + =1266.76.
1H NMR(400MHz,DMSO-d6)δ11.03(s,1H),9.22(m,1H),9.12(s,2H),8.70-8.59(m,1H),8.25-7.89(m,8H),7.81(m,2H),7.54(m,2H),7.49-7.28(m,4H),7.25(t,J＝8.6Hz,2H),5.17(dd,J＝13.3,5.3Hz,1H),4.75-4.37(m,9H),4.31-4.10(m,4H),3.87-3.65(m,2H),3.43(s,3H),3.21(s,1H),3.02-2.86(m,1H),2.73-2.53(m,3H),2.50-2.25(m,11H),2.12-1.90(m,2H),1.90-1.77(m,2H),1.63(m,2H),1.42-1.23(m,8H),0.95-0.75(m,12H). 1 H NMR (400MHz, DMSO-d 6 ) δ11.03(s,1H),9.22(m,1H),9.12(s,2H),8.70-8.59(m,1H),8.25-7.89(m,8H ),7.81(m,2H),7.54(m,2H),7.49-7.28(m,4H),7.25(t,J＝8.6Hz,2H),5.17(dd,J＝13.3,5.3Hz,1H) ,4.75-4.37(m,9H),4.31-4.10(m,4H),3.87-3.65(m,2H),3.43(s,3H),3.21(s,1H),3.02-2.86(m,1H) ,2.73-2.53(m,3H),2.50-2.25(m,11H),2.12-1.90(m,2H),1.90-1.77(m,2H),1.63(m,2H),1.42-1.23(m, 8H),0.95-0.75(m,12H).
实施例3.2 2-(2,6-二氧代哌啶-3-基)-N-(4-((12S,15S)-12-(4-(二丙胺基)丁基)-15-异丙基-22-(2-(甲磺酰基)嘧啶-5-基)-8,11,14,17-四氧代-2-苯乙基-5-氧杂-2,7,10,13,16-五氮杂二十二碳-21-炔-1-基)苄基)-1-氧代异吲哚啉-5-甲酰胺(DL-2)的合成
Example 3.2 2-(2,6-dioxopiperidin-3-yl)-N-(4-((12S,15S)-12-(4-(dipropylamino)butyl)-15-iso Propyl-22-(2-(methanesulfonyl)pyrimidin-5-yl)-8,11,14,17-tetraoxo-2-phenylethyl-5-oxa-2,7,10,13 ,Synthesis of 16-pentaazadococ-21-yn-1-yl)benzyl)-1-oxoisoindoline-5-carboxamide (DL-2)
步骤一：step one:
将化合物L-3(24mg，0.037mmol)，化合物L-2(22mg，0.037mmol)加入反应瓶中，然后加入DMF(1ml)，0℃溶解完全后，再加入DMTMM(13mg，0.044mmol)，继续保持0℃反应1.5h。反应液经制备高效液相色谱纯化(乙腈：水(含0.1％甲酸)＝5％-90％)，得目标化合物DL-2(5mg)。Add compound L-3 (24 mg, 0.037 mmol) and compound L-2 (22 mg, 0.037 mmol) into the reaction bottle, then add DMF (1 ml). After complete dissolution at 0°C, add DMTMM (13 mg, 0.044 mmol). Continue to maintain the reaction at 0°C for 1.5h. The reaction solution was purified by preparative high performance liquid chromatography (acetonitrile: water (containing 0.1% formic acid) = 5%-90%) to obtain the target compound DL-2 (5 mg).
ESI-MS[M+H]+＝1202.6.ESI-MS[M+H] + =1202.6.
1H NMR(400MHz,DMSO-d6)δ11.04(s,1H),9.22(s,1H),9.13(s,2H),8.55(s,1H),8.25(s,1H),8.15(d,J＝11.5Hz,2H),8.05(d,J＝7.2Hz,2H),7.95(d,J＝8.5Hz,1H),7.85(d,J＝7.9Hz,1H),7.29-7.25(m,5H),7.20-7.17(m,3H),5.17(dd,J＝13.2,5.1Hz,1H),4.58-4.50(m,5H),4.27–4.15(m,2H),3.89-3.84(m,2H),3.81-3.78(m,2H),3.75–3.69(m,2H),3.66(s,3H),2.98–2.95(m,2H)，2.77–2.62(m,9H),2.60–2.55(m,3H),2.44–2.27(m,9H),2.10–1.93(m,3H),1.84–1.45(m,2H)，1.42-1.35(m,6H),0.85(dt,J＝12.0,7.4Hz,12H). 1 H NMR (400MHz, DMSO-d 6 ) δ11.04(s,1H),9.22(s,1H),9.13(s,2H),8.55(s,1H),8.25(s,1H),8.15( d,J＝11.5Hz,2H),8.05(d,J＝7.2Hz,2H),7.95(d,J＝8.5Hz,1H),7.85(d,J＝7.9Hz,1H),7.29-7.25( m,5H),7.20-7.17(m,3H),5.17(dd,J＝13.2,5.1Hz,1H),4.58-4.50(m,5H),4.27–4.15(m,2H),3.89-3.84( m,2H),3.81-3.78(m,2H),3.75–3.69(m,2H),3.66(s,3H),2.98–2.95(m,2H),2.77–2.62(m,9H),2.60– 2.55(m,3H),2.44–2.27(m,9H),2.10–1.93(m,3H),1.84–1.45(m,2H),1.42-1.35(m,6H),0.85(dt,J＝12.0 ,7.4Hz,12H).
实施例3.3 4-((S)-2-((S)-2-(6-(2,5-二氧代-2,5-二氢-1H-吡咯-1-基)己酰胺基)-3-甲基丁酰胺基)-5-脲基戊酰胺基)苄基(3,4-二氯苯基乙基)(4-((2-(2,6-二氧代哌啶-3-基)-1-氧代吲哚啉-5-甲酰胺基)甲基)苄基)氨基甲酸酯(DL-3)的合成
Example 3.3 4-((S)-2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoamido) -3-Methylbutylamido)-5-ureidovaleramido)benzyl(3,4-dichlorophenylethyl)(4-((2-(2,6-dioxopiperidine- Synthesis of 3-yl)-1-oxoindoline-5-carboxamido)methyl)benzyl)carbamate (DL-3)
步骤一：化合物DL-3的合成Step 1: Synthesis of compound DL-3
将化合物D-10(160mg,0.28mmol)，Mc-Val-Cit-PAB-PNP(cas No:159857-81-5,224mg,0.30mmol)和二甲基吡啶(59mg,0.55mmol)溶于18mL DMF中，向其中加入HOBt(19mg,0.14mmol)。常温反应48h。向其中加50mL水淬灭反应。将析出的固体抽滤，滤饼用水淋洗。滤饼干燥，经制备纯化得目标化合物DL-3(22mg)。Compound D-10 (160mg, 0.28mmol), Mc-Val-Cit-PAB-PNP (cas No: 159857-81-5, 224mg, 0.30mmol) and lutidine (59mg, 0.55mmol) were dissolved in 18mL DMF , HOBt (19 mg, 0.14 mmol) was added thereto. Reaction at room temperature for 48 hours. Add 50 mL of water to quench the reaction. The precipitated solid is suction filtered, and the filter cake is rinsed with water. The filter cake was dried, and the target compound DL-3 (22 mg) was obtained through preparation and purification.
ESI-MS[M+H]+:1177.5。ESI-MS[M+H] + :1177.5.
1H NMR(400MHz,DMSO-d6)δ11.01(s,1H),9.99(s,1H),9.20(t,J＝5.6Hz,1H),8.11–8.06(m,2H),8.00(d,J＝7.9Hz,1H),7.85–7.77(m,2H),7.63–7.53 (m,2H),7.51–7.02(m,9H),6.99(s,2H),5.97(t,J＝5.7Hz,1H),5.40(s,2H),5.14(dd,J＝13.3,5.1Hz,1H),5.04–4.89(m,2H),4.54–4.32(m,7H),4.23–4.14(m,1H),3.08–2.88(m,3H),2.82–2.69(m,2H),2.66–2.56(m,1H),2.44–2.33(m,2H),2.24–2.09(m,2H),2.07–1.91(m,3H),1.74–1.55(m,2H),1.53–1.33(m,6H),1.22–1.12(m,2H),0.83(dd,J＝12.5,6.8Hz,6H). 1 H NMR (400MHz, DMSO-d 6 ) δ11.01 (s, 1H), 9.99 (s, 1H), 9.20 (t, J = 5.6Hz, 1H), 8.11–8.06 (m, 2H), 8.00 ( d,J＝7.9Hz,1H),7.85–7.77(m,2H),7.63–7.53 (m,2H),7.51–7.02(m,9H),6.99(s,2H),5.97(t,J＝5.7Hz,1H),5.40(s,2H),5.14(dd,J＝13.3,5.1 Hz,1H),5.04–4.89(m,2H),4.54–4.32(m,7H),4.23–4.14(m,1H),3.08–2.88(m,3H),2.82–2.69(m,2H), 2.66–2.56(m,1H),2.44–2.33(m,2H),2.24–2.09(m,2H),2.07–1.91(m,3H),1.74–1.55(m,2H),1.53–1.33(m ,6H),1.22–1.12(m,2H),0.83(dd,J=12.5,6.8Hz,6H).
实施例4.抗PSMA抗体的制备与鉴定Example 4. Preparation and identification of anti-PSMA antibodies
实施例4.1抗PSMA抗体的表达Example 4.1 Expression of anti-PSMA antibodies
将VH氨基酸序列SEQ ID NO:1连接上IgG1的重链恒定区氨基酸序列(SEQ ID NO:2)，通过密码子优化及基因合成(上海百英生物科技有限公司)，然后构建到pcDNA3.4载体上，命名为B21231602H。Connect the VH amino acid sequence SEQ ID NO:1 to the heavy chain constant region amino acid sequence of IgG1 (SEQ ID NO:2), and then construct it into pcDNA3.4 through codon optimization and gene synthesis (Shanghai Baiying Biotechnology Co., Ltd.) On the carrier, it is named B21231602H.
将VL氨基酸序列SEQ ID NO:3连接上IgG1的轻链恒定区氨基酸序列(SEQ ID NO:4)，通过密码子优化及基因合成(上海百英生物科技有限公司)，然后构建到pcDNA3.4载体上，命名为B21231602L。Connect the VL amino acid sequence SEQ ID NO:3 to the light chain constant region amino acid sequence of IgG1 (SEQ ID NO:4), and then construct it into pcDNA3.4 through codon optimization and gene synthesis (Shanghai Baiying Biotechnology Co., Ltd.) On the carrier, it is named B21231602L.
通过PEI max试剂将抗体PSMA抗体表达质粒B21231602H、B21231602L共转染CHO细胞进行表达，将包含孵育后细胞的摇瓶放入37℃，270rpm，8％CO2培养，24h后添加补料/丁酸钠/双抗，继续培养3-7d。收集上清液，通过ProA磁珠进行纯化，获得抗PSMA抗体命名为PSMA mAb(其重链氨基酸序列为SEQ ID NO:5，轻链氨基酸序列为SEQ ID NO:6)。Use PEI max reagent to co-transfect the antibody PSMA antibody expression plasmids B21231602H and B21231602L into CHO cells for expression. Place the shake flask containing the incubated cells into 37°C, 270rpm, and 8% CO2 for culture. Add feed/sodium butyrate after 24 hours. /double antibody, continue culturing for 3-7 days. The supernatant was collected and purified by ProA magnetic beads to obtain an anti-PSMA antibody named PSMA mAb (its heavy chain amino acid sequence is SEQ ID NO: 5 and light chain amino acid sequence is SEQ ID NO: 6).
序列及其具体信息


Sequence and its specific information


实施例4.2抗PSMA抗体结合人PSMA蛋白的亲和力评价Example 4.2 Evaluation of the affinity of anti-PSMA antibodies for binding to human PSMA protein
本实验目的是研究PSMA mAb(由实施例4.1制备的)对人PSMA蛋白的亲和力，为PSMA mAb体内外药效试验提供依据。阴性对照为具有天然的序列的human IgG1。The purpose of this experiment is to study the affinity of PSMA mAb (prepared in Example 4.1) to human PSMA protein and provide a basis for PSMA mAb in vitro and in vivo efficacy testing. The negative control is human IgG1 with native sequence.
用ELISA coating buffer(carbonate buffer,pH 9.5,Biolegend,421701)稀释人PSMA蛋白(购自：Acrobiosystems货号，PSA-H52H3)至0.5ug/ml，100ul/孔包被酶标板，孔包被酶标板，4℃孵育过夜。清洗，即：弃上清，用ELISA wash buffer(购自：Multiscience货号，EK0011)清洗酶标板，300ul/孔，洗三次，将板拍干。100ul/孔向酶标板中加入ELISA assay dilute buffer(购自：biolegend货号421203)，37℃孵育1h。再清洗。用ELISA assay dilute buffer稀释human IgG1和PSMA mAb，最高浓度10ug/ml，随后3倍稀释样品，最后共15个浓度梯。稀释好的样品按照50ul/孔加入酶标板中，37℃孵育2h。再清洗。用ELISA assay dilute buffer按照1:1000稀释羊抗人IgG-HRP抗体(购自：Sino bio货号SSA002)，100ul/孔加入酶标板，37℃孵育1h。再清洗。将TMB显色液A/B液1:1预混，100ul/孔加入酶标板中。1-2min后100ul/孔加入2M H2SO4终止。在 波长450nm和570nm处进行吸收光读数，最终读数为A(450nm-570nm)。使用GraphPad 7.00绘制结合曲线并计算EC50值。Use ELISA coating buffer (carbonate buffer, pH 9.5, Biolegend, 421701) to dilute human PSMA protein (purchased from: Acrobiosystems product number, PSA-H52H3) to 0.5ug/ml, 100ul/well coated enzyme plate, well coated enzyme label Plate and incubate overnight at 4°C. Clean, that is: discard the supernatant, wash the enzyme plate with ELISA wash buffer (purchased from: Multiscience, product number, EK0011), 300ul/well, wash three times, and pat the plate dry. Add 100ul/well of ELISA assay dilute buffer (purchased from: biolegend Cat. No. 421203) to the enzyme plate, and incubate at 37°C for 1 hour. Clean again. Use ELISA assay dilute buffer to dilute human IgG1 and PSMA mAb to a maximum concentration of 10ug/ml, and then dilute the sample 3 times, resulting in a total of 15 concentration gradients. Add the diluted sample to the enzyme plate at 50ul/well and incubate at 37°C for 2 hours. Clean again. Use ELISA assay dilute buffer to dilute the goat anti-human IgG-HRP antibody (purchased from: Sino bio Cat. No. SSA002) at 1:1000, add 100ul/well to the enzyme plate, and incubate at 37°C for 1 hour. Clean again. Premix TMB chromogenic solution A/B solution 1:1, and add 100ul/well to the enzyme plate. After 1-2 minutes, add 100ul/well of 2M H 2 SO 4 to terminate. exist Absorption light readings were taken at wavelengths 450nm and 570nm, and the final reading was A (450nm-570nm). Binding curves were drawn and EC50 values calculated using GraphPad 7.00.
结果显示PSMA mAb与人PSMA蛋白有很好的结合活性，阴性对照human IgG1不结合人PSMA。根据结合曲线拟合计算得到EC50，PSMA mAb对人PSMA蛋白的EC50值为6.67nM。The results show that PSMA mAb has good binding activity to human PSMA protein, and the negative control human IgG1 does not bind to human PSMA. The EC50 was calculated based on binding curve fitting. The EC50 value of PSMA mAb for human PSMA protein was 6.67nM.
实施例4.3抗PSMA抗体的内吞活性检测(信号消失法)Example 4.3 Endocytic activity detection of anti-PSMA antibodies (signal disappearance method)
本实验目的是研究PSMA mAb(Progenics)在22RV细胞中的内吞活性，为PSMA mAb(Progenics)体内外药效试验提供依据。The purpose of this experiment is to study the endocytic activity of PSMA mAb (Progenics) in 22RV cells and provide a basis for the in vivo and in vitro efficacy testing of PSMA mAb (Progenics).
将5×106 22RV1细胞(美森，CTCC-400-0228)(2mL的10％FBS的DMEM)接种到6孔板中，在37℃、5％CO2条件下过夜培养。第二日，去除细胞上清后，用冷的PBS稀释PMSA抗体至50ug/mL后，均匀加入22RV1细胞中，并在4℃孵育30min。去除孵育液体后，加入2mL的10％FBS的DMEM，分别在37℃、5％CO2条件下孵育0h、2h、4h、8h、18h，并在每个时间点收集细胞样本后，用多聚甲醛固定。冷的PBS洗2-3遍后，加入二抗anti-human IgG-PE(Abcam，ab98596)(1:1000)孵育30min，细胞送流式细胞仪检测(FACS)。在本实验中，用PSMA抗体的内化率来代表抗体的内吞活性。即，内化率％＝[1-(MFI0H–MFIXH)]*100；X为孵育时间。内化率越高，抗体的内吞活性越好。5×10 6 22RV1 cells (Mason, CTCC-400-0228) (2 mL of DMEM with 10% FBS) were seeded into a 6-well plate and cultured overnight at 37°C, 5% CO2 . On the next day, after removing the cell supernatant, dilute the PMSA antibody to 50ug/mL with cold PBS, add it evenly to the 22RV1 cells, and incubate at 4°C for 30 minutes. After removing the incubation liquid, add 2 mL of DMEM with 10% FBS and incubate at 37°C and 5% CO2 for 0h, 2h, 4h, 8h, and 18h respectively. After collecting cell samples at each time point, use poly Formaldehyde fixation. After washing 2-3 times with cold PBS, add secondary antibody anti-human IgG-PE (Abcam, ab98596) (1:1000) and incubate for 30 minutes, and send the cells to flow cytometry (FACS). In this experiment, the internalization rate of PSMA antibody was used to represent the endocytic activity of the antibody. That is, internalization rate % = [1-(MFI 0H – MFI XH )]*100; X is the incubation time. The higher the internalization rate, the better the endocytic activity of the antibody.
FACS结果显示，PSMA抗体随着孵育时间的延长，而表现出递增的内化率，即内吞活性随着孵育时间延长而增强，最终达到饱和状态。根据上述计算公式，计算得到内化率％，即，PSMA抗体与22RV1细胞共孵育18h，其内化率为60％。FACS results showed that the PSMA antibody showed an increasing internalization rate as the incubation time prolonged, that is, the endocytic activity increased with the prolongation of the incubation time, and finally reached a saturation state. According to the above calculation formula, the internalization rate % was calculated, that is, when the PSMA antibody was incubated with 22RV1 cells for 18 hours, the internalization rate was 60%.
实施例5.抗体药物偶联物(ADC)的合成Example 5. Synthesis of Antibody Drug Conjugates (ADC)
实施例5.1：ADC-001的制备Example 5.1: Preparation of ADC-001
取0.5mL实施例4.1获得的PSMA mAb(8.84mg/mL)，用0.005mL 20mM PB+100mM依地酸二钠溶液(pH 7.6)稀释，以0.5M Na2HPO4溶液调pH至7.45，加入20mM TCEP(三(2-羧乙基)膦，0.0082ml，0.164μmol)溶液混匀，室温放置90min。向上述溶液加入DL-1(0.470mg，12倍抗体物质摩尔数量)的二甲基亚砜(0.0558mL)溶液，混匀，室温静置2h，完毕后采用离心超滤管(Merck,Amicon Ultra-15)进行换液，缓冲液置换为pH＝5.9的20mM His-Hcl缓冲溶液。得到DL-1与PSMA mAb的偶联产物ADC-001。质谱法测定DAR值为8.0。 Take 0.5mL of PSMA mAb (8.84mg/mL) obtained in Example 4.1, dilute it with 0.005mL of 20mM PB+100mM disodium edetate solution (pH 7.6), adjust the pH to 7.45 with 0.5M Na 2 HPO 4 solution, and add Mix 20mM TCEP (tris(2-carboxyethyl)phosphine, 0.0082ml, 0.164μmol) solution and leave it at room temperature for 90 minutes. Add a solution of DL-1 (0.470 mg, 12 times the molar amount of antibody material) in dimethyl sulfoxide (0.0558 mL) to the above solution, mix well, and let stand at room temperature for 2 hours. After completion, use a centrifugal ultrafiltration tube (Merck, Amicon Ultra) -15) Change the liquid, and replace the buffer with a 20mM His-Hcl buffer solution with pH=5.9. The coupling product ADC-001 of DL-1 and PSMA mAb was obtained. The DAR value determined by mass spectrometry was 8.0.
DAR值测定方法如下：The method for measuring DAR value is as follows:
样品处理：Sample processing:
各取ADC样品50μg，用超纯水稀释到0.5mg/ml，然后加入DTT(1μl，1M)混匀后离心取上清进样。Take 50 μg of each ADC sample, dilute it to 0.5 mg/ml with ultrapure water, then add DTT (1 μl, 1M), mix well, and centrifuge to take the supernatant for injection.
仪器信息：
Instrument information:
液相参数：
Liquid phase parameters:
质谱参数：

Mass spectrometry parameters:

实验结果：

Experimental results:

实施例5.2：ADC-002的制备 Example 5.2: Preparation of ADC-002
采用与实施例5.1类似的操作，用DL-2替换DL-1，得到DL-2与PSMA mAb的偶联产物ADC-002。质谱法测定DAR值为8.0。

Using a similar operation to Example 5.1, replacing DL-1 with DL-2, ADC-002, the coupling product of DL-2 and PSMA mAb, was obtained. The DAR value determined by mass spectrometry was 8.0.

表中，LC代表抗体轻链；HC代表抗体重链；DAR1代表包含轻链或重链偶联1个毒素连接子的偶联物；DAR2代表包含轻链或重链偶联2个毒素连接子的偶联物；DAR3代表包含轻链或重链偶联3个毒素连接子的偶联物。下文中LC、HC、DAR1、DAR2、DAR3如上说明。In the table, LC represents the antibody light chain; HC represents the antibody heavy chain; DAR1 represents the conjugate containing a light chain or heavy chain coupled to a toxin linker; DAR2 represents a conjugate containing a light chain or heavy chain coupled to two toxin linkers. Conjugates; DAR3 represents conjugates containing three toxin linkers coupled to a light or heavy chain. In the following, LC, HC, DAR1, DAR2, and DAR3 are as described above.
测定PSMA抗体轻链偶联0～1个毒素连接子(LC，DAR1比例分别为0.0％，100.0％)、重链偶联0～3个毒素连接子(HC、DAR1、DAR2、DAR3的比例分别为0％，0％，0％，100％)，由此计算ADC-001的DAR值(抗体-药物偶联比)为8.0。Determine the proportion of the PSMA antibody light chain coupled to 0 to 1 toxin linkers (LC, DAR1 ratios are 0.0%, 100.0%, respectively), and the heavy chain coupled to 0 to 3 toxin linkers (HC, DAR1, DAR2, DAR3, respectively). is 0%, 0%, 0%, 100%), thus the DAR value (antibody-drug conjugation ratio) of ADC-001 is calculated to be 8.0.
实施例5.3：ADC-003的制备
Example 5.3: Preparation of ADC-003
采用与实施例5.1类似的操作，用DL-3替换DL-1，得到DL-3与PSMA mAb的偶联产物ADC-003。质谱法测定DAR值为8.2。
Using a similar operation to Example 5.1, replacing DL-1 with DL-3, ADC-003, the coupling product of DL-3 and PSMA mAb, was obtained. The DAR value determined by mass spectrometry was 8.2.
实施例6.生物活性测试Example 6. Biological activity testing
实施例6.1：蛋白降解剂类生物活性化合物对c-Myc和GSPT1蛋白降解活性测试Example 6.1: Test of protein degradation activity of protein-degrading bioactive compounds on c-Myc and GSPT1 proteins
1.实验材料1. Experimental materials
细胞培养条件：本发明实施例中细胞均购买自中国科学院上海细胞库。其中人急性早幼粒白血病细胞(HL60细胞)培养在RPMI-1640(meilunbio，MA0215)+20％FBS(厂家：BI,货号：04-001-1ACS)+1％青霉素/链霉素(厂家：Hyclone，货号：SH40003.01)的培养基中。细胞培养条件均为5％CO2浓度与37％湿度。待培养至覆盖率达80％左右时，按照1:3比例传代。Cell culture conditions: The cells in the examples of the present invention were all purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences. Among them, human acute promyelocytic leukemia cells (HL60 cells) were cultured in RPMI-1640 (meilunbio, MA0215) + 20% FBS (manufacturer: BI, product number: 04-001-1ACS) + 1% penicillin/streptomycin (manufacturer: Hyclone, Cat. No.: SH40003.01) culture medium. Cell culture conditions were all 5% CO2 concentration and 37% humidity. When the coverage rate reaches about 80%, the cells are passaged in a ratio of 1:3.
2.细胞药物处理和蛋白质提取2. Cell drug processing and protein extraction
2.1细胞处理2.1 Cell processing
HL60细胞在指数生长期时，将HL60细胞铺板至6孔板中，每孔1×106个细胞，加入本发明化合物对细胞进行培养，药物处理6h后提取蛋白质。When the HL60 cells are in the exponential growth phase, the HL60 cells are plated into a 6-well plate with 1×10 6 cells per well. The compound of the present invention is added to culture the cells, and the protein is extracted after 6 hours of drug treatment.
2.2细胞蛋白质提取2.2 Cell protein extraction
将细胞收集于1.5mL离心管中，1000rmp离心5分钟后弃去上清，加入1ml的PBS清洗细胞，1000rmp离心5分钟后，弃去上清，每管中加入100μL RIPA裂解液(Solarbio，货号R0010)(含100μM PMSF)，充分混合后冰上静置30min，之后用离心机4℃12000rpm离心20min，取上清液用于Western blotting(WB)实验，样品可放于-80℃保存。 Collect the cells in a 1.5 mL centrifuge tube, centrifuge at 1000 rpm for 5 minutes, discard the supernatant, add 1 ml of PBS to wash the cells, centrifuge at 1000 rpm for 5 minutes, discard the supernatant, and add 100 μL RIPA lysis buffer (Solarbio, Cat. No. R0010) (containing 100 μM PMSF), mix thoroughly and let stand on ice for 30 minutes, then centrifuge at 12000 rpm at 4°C for 20 minutes, and take the supernatant for Western blotting (WB) experiments. The sample can be stored at -80°C.
2.3蛋白质浓度测定2.3 Determination of protein concentration
使用BCA蛋白浓度测定试剂盒(来自于Thermo Fisher，货号为23225)，按照下表1配制BSA标准测定溶液和上述步骤2.2中得到的用于Western blotting(WB)实验的上清液(上清液可稀释后检测)，使用96孔板加样，各孔用PBS补足至20μl后，分别加入200μl BCA工作液(按试剂盒配制)，混匀后于60℃放置10min后于562nm处检测吸光度，记录读数后以标准品浓度梯度作标准曲线，代入样品吸光度计算样品蛋白质浓度。Use the BCA protein concentration determination kit (from Thermo Fisher, Cat. No. 23225) to prepare the BSA standard determination solution and the supernatant (supernatant) obtained in the above step 2.2 for the Western blotting (WB) experiment according to Table 1 below. (can be diluted and tested), use a 96-well plate to add samples, add PBS to each well to 20μl, add 200μl BCA working solution (prepared according to the kit), mix well and place it at 60°C for 10 minutes, then detect the absorbance at 562nm. After recording the readings, use the standard concentration gradient to make a standard curve, and substitute the sample absorbance to calculate the sample protein concentration.
表1蛋白定量标准品配制
Table 1 Preparation of protein quantitative standards
3Western blotting实验标准流程3Western blotting experimental standard procedure
3.1蛋白质变性：取RIPA蛋白裂解液，加入5×Loading Buffer(SDS，甘油，溴酚蓝，TRIS，购买于Solarbio，货号P1040)，100℃变性5min。3.1 Protein denaturation: Take RIPA protein lysate, add 5× Loading Buffer (SDS, glycerol, bromophenol blue, TRIS, purchased from Solarbio, product number P1040), and denature at 100°C for 5 minutes.
3.2上样及电泳：使用10％的15孔预制胶(金斯瑞，货号M00666)及对应电泳液(金斯瑞，货号M00138)，每孔上样蛋白protein ladder(购买于thermo，货号26617)以及同质量蛋白样品之后，200V电泳30min。3.2 Loading and electrophoresis: Use 10% 15-well precast gel (GenScript, Cat. No. M00666) and corresponding electrophoresis solution (GenScript, Cat. No. M00138), and load protein ladder (purchased from Thermo, Cat. No. 26617) into each well. After protein samples of the same mass were electrophoresed at 200V for 30 minutes.
3.3转膜：将胶取下后切下多余部分，通过湿转法转移到PVDF膜(Millipore，货号ISEQ00010)上(PVDF膜需要先用甲醇活化1min后使用)，300mA，2h，转膜过程中大量产热，需要用冰盒降温。3.3 Membrane transfer: Remove the glue and cut off the excess part, and transfer it to PVDF membrane (Millipore, Cat. No. ISEQ00010) by wet transfer method (PVDF membrane needs to be activated with methanol for 1 minute before use), 300mA, 2h, during the transfer process Generates a lot of heat and requires an ice box to cool down.
3.4封闭：将转膜结束的PVDF膜放于5％(w/w)脱脂牛奶中室温摇晃封闭1h。3.4 Blocking: Place the transferred PVDF membrane in 5% (w/w) skim milk and shake at room temperature for 1 hour.
3.5孵育一抗：将PVDF膜按照marker上指示分子量大小分别剪开，分别置于c-myc(购买于abcam，货号ab32072)、GSPT1(购买于abcam，货号ab234433)和GAPDH(购买于CST，货号#97166)一抗中，抗体按照1:1000比例用TBST缓冲液(TRIS,KCL和Nacl配制的溶液，用盐酸调节至PH7.4,加入吐温20)稀释，4℃摇床封闭过夜。 3.5 Incubate primary antibodies: Cut the PVDF membrane according to the molecular weight indicated on the marker, and place it on c-myc (purchased from abcam, product number ab32072), GSPT1 (purchased from abcam, product number ab234433) and GAPDH (purchased from CST, product number ab234433) #97166) primary antibody, dilute the antibody with TBST buffer (a solution prepared with TRIS, KCL and Nacl, adjust to pH 7.4 with hydrochloric acid, add Tween 20) at a ratio of 1:1000, and block on a shaker at 4°C overnight.
3.6孵育二抗：孵育一抗后的PVDF膜在摇床上用TBST清洗3次，每次10min，清洗后将膜分别置于(Anti-mouse IgG,HRP-linked Antibody(购买于CST，货号#7076)或Anti-rabbit IgG,HRP-linked Antibody(购买于CST,货号#7074)摇床振荡室温孵育1h。3.6 Incubate secondary antibodies: After incubating the primary antibodies, the PVDF membrane was washed three times with TBST on a shaker, 10 minutes each time. After cleaning, the membranes were placed in (Anti-mouse IgG, HRP-linked Antibody (purchased from CST, Cat. No. #7076 ) or Anti-rabbit IgG, HRP-linked Antibody (purchased from CST, Cat. No. #7074) and incubate at room temperature for 1 hour with shaking on a shaker.
3.7洗膜及曝光：孵育二抗结束后将膜放于TBST中摇床振荡清洗3次，每次10min，洗膜后使用增强化学发光法(ECL)显示化学光的强弱。3.7 Membrane washing and exposure: After incubating the secondary antibody, place the membrane in TBST and shake it with shaking for 3 times, 10 minutes each time. After washing the membrane, use enhanced chemiluminescence (ECL) to display the intensity of chemical light.
按照上面方法，分别对上述细胞中的蛋白降解能力进行了测试，结果如图1所示。其中，Blank组代表未加待测化合物的DMSO空白组。图1显示化合物D-2和化合物D-4在较低浓度(3-10nM)时，即能将HL60细胞中的c-Myc和GSPT1蛋白降解完全。According to the above method, the protein degradation ability in the above cells was tested respectively, and the results are shown in Figure 1. Among them, the Blank group represents the DMSO blank group without adding the test compound. Figure 1 shows that compound D-2 and compound D-4 can completely degrade c-Myc and GSPT1 proteins in HL60 cells at lower concentrations (3-10 nM).
实施例6.2：蛋白降解剂类生物活性化合物对HL60细胞抑制活性检测Example 6.2: Detection of inhibitory activity of protein-degrading bioactive compounds on HL60 cells
活性测试方法：Activity test method:
1.细胞铺板1. Cell plating
将处于对数生长期的HL60细胞计数，并均匀铺在96孔透明底白板中，细胞数量为每孔20000个，每孔100μL。Count the HL60 cells in the logarithmic growth phase and spread them evenly in a 96-well transparent bottom white plate. The number of cells is 20,000 per well and 100 μL per well.
2.细胞加药2. Cell dosing
称取一定量待测化合物，用DMSO稀释至浓度为10mM的母液。取细胞对应的培养基，将待测化合物稀释至0.3-2222.2nM的浓度，并轻轻振荡混匀。将稀释好的化合物梯度稀释液每孔取50μL加入到铺好的细胞中，对照组加入50μL0.1％DMSO，均轻轻振荡混匀。放入细胞培养箱，孵育3天。Weigh a certain amount of the compound to be tested and dilute it with DMSO to a mother solution with a concentration of 10mM. Take the culture medium corresponding to the cells, dilute the compound to be tested to a concentration of 0.3-2222.2nM, and shake gently to mix. Add 50 μL of the diluted compound gradient dilution to each well of the plated cells, and add 50 μL of 0.1% DMSO to the control group, and mix gently by shaking. Place into cell culture incubator and incubate for 3 days.
3.CTG检测3.CTG detection
3.1提前将CellTiter-GloTM(CTG)试剂从-20℃冰箱中取出避光平衡至室温。3.1 Take the CellTiter-Glo TM (CTG) reagent out of the -20°C refrigerator in advance to avoid light and equilibrate it to room temperature.
3.2 96孔板取出，显微镜镜检。看生长情况。3.2 Take out the 96-well plate and inspect it under a microscope. Look at the growth.
3.3取出的96孔板板底用白纸贴上，之后用锡箔纸包住，于37℃、300r，震荡摇匀10min。 3.3 Paste the bottom of the taken out 96-well plate with white paper, then wrap it with tin foil, and shake well for 10 minutes at 37°C and 300r.
3.4每孔加入50ul CTG检测试剂，酶标仪检测Luminescence信号，计算抑制率。通过XLfit软件计算化合物IC50值，评估HL60细胞抑制增殖活性，实验结果如表2所示：3.4 Add 50ul CTG detection reagent to each well, detect the Luminescence signal with a microplate reader, and calculate the inhibition rate. The compound IC 50 value was calculated by XLfit software to evaluate the inhibitory activity of HL60 cells. The experimental results are shown in Table 2:
表2化合物对HL60细胞的抑制增殖活性
Table 2 Inhibitory activity of compounds on HL60 cells
测试结果表明，所测试化合物均对HL60细胞有强效的增殖抑制活性。The test results showed that all the tested compounds had potent proliferation inhibitory activity on HL60 cells.
申请人针对D-1、D-4、D-10、ADC-001、ADC-002、ADC-003对HL60细胞的抑制增殖活性在同一组实验下进行了测试，结果如表3所示：The applicant tested the anti-proliferation activity of D-1, D-4, D-10, ADC-001, ADC-002 and ADC-003 on HL60 cells under the same set of experiments. The results are shown in Table 3:
表3药物对HL60细胞的抑制增殖活性
Table 3 Drug inhibitory activity on HL60 cells
结果表明，相比于小分子，ADC分子降低了对PSMA阴性细胞的杀伤。The results showed that ADC molecules reduced the killing of PSMA-negative cells compared with small molecules.
实施例6.3：化合物或ADC对体外22RV1细胞抑制活性检测Example 6.3: Detection of inhibitory activity of compounds or ADC on 22RV1 cells in vitro
活性测试方法：Activity test method:
1.细胞铺板1. Cell plating
将处于对数生长期的22RV1(中国科学院细胞库，TCHu100)细胞计数，并均匀铺在96孔透明底白板中，细胞数量为每孔10000-20000个，每孔100μL。 Count the 22RV1 (Cell Bank of the Chinese Academy of Sciences, TCHu100) cells in the logarithmic growth phase and spread them evenly in a 96-well transparent bottom white plate. The number of cells is 10,000-20,000 per well, 100 μL per well.
2.化合物或ADC稀释2. Compound or ADC dilution
利用细胞培养对应的培养基(RPMI(Meilun Bio,MA 0215),含10％FBS(gibco,16000-044)和1％青霉素/链霉素(Hyclone,SV30010))，将待测化合物或ADC系列稀释：从浓度30μM起始，3倍稀释，9个浓度，并轻轻振荡混匀。Use the corresponding culture medium for cell culture (RPMI (Meilun Bio, MA 0215), containing 10% FBS (gibco, 16000-044) and 1% penicillin/streptomycin (Hyclone, SV30010)), and add the test compound or ADC series Dilution: starting from a concentration of 30 μM, dilute 3 times to 9 concentrations, and mix gently by shaking.
3.细胞加药3. Cell dosing
将稀释好的待测化合物或ADC梯度稀释液每孔取50μl加入到铺好的细胞中，对照组加入50μl 0.1％DMSO，即药物作用终浓度为10μM起始，3倍稀释，9个浓度。轻轻振荡混匀，放入细胞培养箱，孵育72h。Add 50 μl of the diluted compound to be tested or ADC gradient dilution from each well to the plated cells, and add 50 μl of 0.1% DMSO to the control group, that is, the final concentration of the drug is starting from 10 μM, diluted 3 times, and 9 concentrations. Shake gently to mix, place in a cell culture incubator, and incubate for 72 hours.
4.CTG检测4.CTG detection
将CellTiter-GloTM(CTG)试剂从-20℃冰箱中取出避光平衡至室温。每孔加入50μl，室温避光300rpm震荡20min，酶标仪检测Luminescence信号，计算抑制率IC50，其中小分子蛋白降解药物和ADC药物的试验结果如表4所示：Take the CellTiter-Glo TM (CTG) reagent out of the -20°C refrigerator and equilibrate it to room temperature in the dark. Add 50 μl to each well, shake at 300 rpm for 20 minutes at room temperature in the dark, and detect the Luminescence signal with a microplate reader. Calculate the inhibition rate IC 50 . The test results of small molecule protein degradation drugs and ADC drugs are shown in Table 4:
表4药物对22RV1细胞的抗增殖活性
Table 4 Antiproliferative activity of drugs on 22RV1 cells
测试结果表明，本发明公开的ADC分子具有肿瘤细胞杀伤作用，且相对于小分子本身，制备成ADC后显著提高了抗增殖活性。The test results show that the ADC molecules disclosed in the present invention have tumor cell killing effect, and compared with the small molecules themselves, the anti-proliferative activity after being prepared into ADC is significantly improved.
实施例6.4：ADC药物对22RV1细胞中的c-Myc蛋白降解试验Example 6.4: ADC drug degradation test on c-Myc protein in 22RV1 cells
1.实验材料1. Experimental materials
细胞培养条件：本实施例中细胞均购买自中国科学院上海细胞库。其中人前列腺癌细胞(22RV-1细胞)培养在RPMI-1640(meilunbio，MA0215)+10％FBS(厂家：BI,货号：04-001-1ACS)+1％青霉素/链霉素(厂家：Hyclone，货号：SH40003.01)的培养基中。细胞培养条件均为5％CO2浓度、37％湿度。待培养至覆盖率达80％左右时，按照1:3比例传代。Cell culture conditions: The cells in this example were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences. Among them, human prostate cancer cells (22RV-1 cells) were cultured in RPMI-1640 (meilunbio, MA0215) + 10% FBS (manufacturer: BI, product number: 04-001-1ACS) + 1% penicillin/streptomycin (manufacturer: Hyclone , Catalog No.: SH40003.01) in culture medium. Cell culture conditions were all 5% CO2 concentration and 37% humidity. When the coverage rate reaches about 80%, the cells are passaged in a ratio of 1:3.
2.细胞药物处理和蛋白质提取 2. Cell drug processing and protein extraction
2.1细胞处理2.1 Cell processing
22RV-1细胞在指数生长期时，将22RV-1细胞铺板至6孔板中，每孔1×106个细胞，加入本发明ADC药物对细胞进行培养，药物处理6h后提取蛋白质。When the 22RV-1 cells are in the exponential growth phase, the 22RV-1 cells are plated into a 6-well plate, with 1×10 6 cells per well. The ADC drug of the present invention is added to culture the cells, and the protein is extracted after 6 hours of drug treatment.
2.2细胞蛋白质提取2.2 Cell protein extraction
将细胞收集于1.5mL离心管中，1000rmp离心5分钟后弃去上清，加入1ml的PBS清洗细胞，1000rmp离心5分钟后，弃去上清，每管中加入100μL RIPA裂解液(Solarbio，货号R0010)(含100μM PMSF)，充分混合后冰上静置30min，之后用离心机4℃12000rpm离心20min，取上清液用于Western blotting(WB)实验，样品可放于-80℃保存。Collect the cells in a 1.5 mL centrifuge tube, centrifuge at 1000 rpm for 5 minutes, discard the supernatant, add 1 ml of PBS to wash the cells, centrifuge at 1000 rpm for 5 minutes, discard the supernatant, and add 100 μL RIPA lysis buffer (Solarbio, Cat. No. R0010) (containing 100μM PMSF), mix thoroughly and let stand on ice for 30 minutes, then centrifuge at 12000rpm at 4℃ for 20min, and take the supernatant for Western blotting (WB) experiment. The sample can be stored at -80℃.
2.3蛋白质浓度测定2.3 Determination of protein concentration
使用BCA蛋白浓度测定试剂盒(来自于Thermo Fisher，货号为23225)，按照实施例6.1中表1配制BSA标准测定溶液和步骤2.2中得到的用于Western blotting(WB)实验的上清液(上清液可稀释后检测)，使用96孔板加样，各孔用PBS补足至20μl后，分别加入200μl BCA工作液(按试剂盒配制)，混匀后于60℃放置10min后于562nm处检测吸光度，记录读数后以标准品浓度梯度作标准曲线，代入样品吸光度计算样品蛋白质浓度。Use the BCA protein concentration determination kit (from Thermo Fisher, Cat. No. 23225), prepare the BSA standard determination solution and the supernatant for Western blotting (WB) experiment obtained in step 2.2 according to Table 1 in Example 6.1 (upper The clear solution can be diluted and tested), use a 96-well plate to add samples, make up each well to 20μl with PBS, then add 200μl BCA working solution (prepared according to the kit), mix well and place it at 60°C for 10 minutes before detecting at 562nm. Absorbance, after recording the reading, use the standard concentration gradient to make a standard curve, and substitute it into the sample absorbance to calculate the sample protein concentration.
3 Western blotting实验标准流程3 Western blotting experimental standard procedure
3.1蛋白质变性：取RIPA蛋白裂解液，加入5×Loading Buffer(SDS，甘油，溴酚蓝，TRIS，购买于Solarbio，货号P1040)，100℃变性5min。3.1 Protein denaturation: Take RIPA protein lysate, add 5× Loading Buffer (SDS, glycerol, bromophenol blue, TRIS, purchased from Solarbio, product number P1040), and denature at 100°C for 5 minutes.
3.2上样及电泳：使用10％的15孔预制胶(金斯瑞，货号M00666)及对应电泳液(金斯瑞，货号M00138)，每孔上样蛋白protein ladder(购买于thermo，货号26617)以及同质量蛋白样品之后，200V电泳30min。3.2 Loading and electrophoresis: Use 10% 15-well precast gel (GenScript, Cat. No. M00666) and corresponding electrophoresis solution (GenScript, Cat. No. M00138), and load protein ladder (purchased from Thermo, Cat. No. 26617) into each well. After protein samples of the same mass were electrophoresed at 200V for 30 minutes.
3.3转膜：将胶取下后切下多余部分，通过湿转法转移到PVDF膜(Millipore，货号ISEQ00010)上(PVDF膜需要先用甲醇活化1min后使用)，300mA，2h，转膜过程中大量产热，需要用冰盒降温。3.3 Membrane transfer: Remove the glue and cut off the excess part, and transfer it to PVDF membrane (Millipore, Cat. No. ISEQ00010) by wet transfer method (PVDF membrane needs to be activated with methanol for 1 minute before use), 300mA, 2h, during the transfer process Generates a lot of heat and requires an ice box to cool down.
3.4封闭：将转膜结束的PVDF膜放于5％(w/w)脱脂牛奶中室温摇晃封闭1h。3.4 Blocking: Place the transferred PVDF membrane in 5% (w/w) skim milk and shake at room temperature for 1 hour.
3.5孵育一抗：将PVDF膜按照marker上指示分子量大小分别剪开，分别置于c-myc(购买于abcam，货号ab32072)和β-actin(购买于CST，货号3700S) 一抗中，抗体按照1:1000比例用TBST缓冲液(TRIS,KCl和NaCl配制的溶液，用盐酸调节至PH7.4,加入吐温20)稀释，4℃摇床封闭过夜。3.5 Incubate the primary antibody: Cut the PVDF membrane according to the molecular weight indicated on the marker, and place them respectively in c-myc (purchased from abcam, product number ab32072) and β-actin (purchased from CST, product number 3700S) In the primary antibody, the antibody was diluted with TBST buffer (a solution prepared with TRIS, KCl and NaCl, adjusted to pH 7.4 with hydrochloric acid, and Tween 20 was added) at a ratio of 1:1000, and blocked overnight on a shaker at 4°C.
3.6孵育二抗：孵育一抗后的PVDF膜在摇床上用TBST清洗3次，每次10min，清洗后将膜分别置于(Anti-mouse IgG,HRP-linked Antibody(购买于CST，货号#7076)或Anti-rabbit IgG,HRP-linked Antibody(购买于CST,货号#7074)摇床振荡室温孵育1h。3.6 Incubate secondary antibodies: After incubating the primary antibodies, the PVDF membrane was washed three times with TBST on a shaker, 10 minutes each time. After cleaning, the membranes were placed in (Anti-mouse IgG, HRP-linked Antibody (purchased from CST, Cat. No. #7076 ) or Anti-rabbit IgG, HRP-linked Antibody (purchased from CST, Cat. No. #7074) and incubate at room temperature for 1 hour with shaking on a shaker.
3.7洗膜及曝光：孵育二抗结束后将膜放于TBST中摇床振荡清洗3次，每次10min，洗膜后使用增强化学发光法(ECL)显示化学光的强弱。3.7 Membrane washing and exposure: After incubating the secondary antibody, place the membrane in TBST and shake it with shaking for 3 times, 10 minutes each time. After washing the membrane, use enhanced chemiluminescence (ECL) to display the intensity of chemical light.
试验结果如图2所示(其中Ctrl是指步骤2.1细胞处理中以溶剂替代药物，即溶剂对照组)，结果显示，所制备的ADC能显著降解22RV1细胞中的c-Myc蛋白。The test results are shown in Figure 2 (where Ctrl refers to the use of solvent instead of drug in step 2.1 cell treatment, that is, the solvent control group). The results show that the prepared ADC can significantly degrade the c-Myc protein in 22RV1 cells.
实施例6.5化合物或ADC对体外hPBMC和Vcap细胞抑制活性检测Example 6.5 Detection of inhibitory activity of compounds or ADC on hPBMC and Vcap cells in vitro
活性测试方法：Activity test method:
1.细胞铺板1. Cell plating
将处于对数生长期的人外周血单个核细胞hPBMCs(购买于上海澳能生物技术有限公司，培养基为RPMI1640(Meilun Bio,MA 0215),含10％FBS(gibco,16000-044)和1％青霉素/链霉素(Hyclone,SV30010)；人前列腺癌细胞Vcap(购买自ATCC，培养基为DMEM(Hyclone，货号：SH30022.01)+10％FBS(厂家：BI,货号：04-001-1ACS)+1％青霉素/链霉素(厂家：Hyclone，货号：SH40003.01)、细胞计数，并均匀铺在96孔透明底白板中，细胞数量为每孔5000～10000个，每孔100μL。Human peripheral blood mononuclear cells hPBMCs in the logarithmic growth phase (purchased from Shanghai Aoneng Biotechnology Co., Ltd., the culture medium is RPMI1640 (Meilun Bio, MA 0215), containing 10% FBS (gibco, 16000-044) and 1 % penicillin/streptomycin (Hyclone, SV30010); human prostate cancer cell Vcap (purchased from ATCC, the culture medium is DMEM (Hyclone, product number: SH30022.01) + 10% FBS (manufacturer: BI, product number: 04-001- 1ACS) + 1% penicillin/streptomycin (Manufacturer: Hyclone, Catalog No.: SH40003.01), count the cells, and spread them evenly in a 96-well transparent bottom white plate. The number of cells is 5,000 to 10,000 per well, 100 μL per well.
2.化合物或ADC稀释2. Compound or ADC dilution
利用细胞培养对应的培养基将待测化合物或ADC系列稀释：从浓度1μM起始，3倍稀释，9个浓度，并轻轻振荡混匀。Use the culture medium corresponding to the cell culture to serially dilute the compound to be tested or ADC: starting from a concentration of 1 μM, diluting 3 times to 9 concentrations, and shaking gently to mix.
3.细胞加药3. Cell dosing
将稀释好的待测化合物或ADC梯度稀释液每孔取50μl加入到铺好的细胞中，对照组加入50μl 0.1％DMSO，即药物作用终浓度为1μM起始，3倍稀释，9个浓度。轻轻振荡混匀，放入细胞培养箱，孵育72h。Add 50 μl of the diluted compound to be tested or ADC gradient dilution from each well to the plated cells, and add 50 μl of 0.1% DMSO to the control group, that is, the final drug concentration is 1 μM starting from 1 μM, diluted 3 times, and 9 concentrations. Shake gently to mix, place in a cell culture incubator, and incubate for 72 hours.
4.CTG检测 4.CTG detection
将CellTiter-GloTM(CTG,Vazyme，货号：DD1102-03)试剂从-20℃冰箱中取出避光平衡至室温。每孔加入50μl，室温避光300rpm震荡20min，使用Biotek多功能酶标仪(型号：Syneray H1Hybrid Reader)检测Luminescence信号，计算抑制率IC50，其中小分子蛋白降解药物和ADC药物的试验结果如表5和6所示：Take the CellTiter-Glo TM (CTG, Vazyme, Cat. No.: DD1102-03) reagent from the -20°C refrigerator to avoid light and equilibrate to room temperature. Add 50 μl to each well, shake at 300 rpm for 20 minutes at room temperature in the dark, and use a Biotek multifunctional microplate reader (Model: Syneray H1Hybrid Reader) to detect the Luminescence signal and calculate the inhibition rate IC 50. The test results of small molecule protein degradation drugs and ADC drugs are as shown in the table 5 and 6 shown:
表5药物对hPBMC细胞的抑制增殖活性
Table 5 Inhibitory activity of drugs on hPBMC cells
综合表3和表5的实验结果，相对于小分子化合物本身，ADC对PSMA阴性细胞HL60细胞和正常人类细胞hPBMC细胞的抑制增殖活性减低，，制备成ADC之后大大提高了治疗窗口。Based on the experimental results in Table 3 and Table 5, compared with the small molecule compound itself, ADC has reduced inhibitory activity on PSMA-negative HL60 cells and normal human cell hPBMC cells. After being prepared into ADC, the therapeutic window is greatly improved.
表6药物对Vcap细胞的抑制增殖活性
Table 6: Antiproliferative activity of drugs on Vcap cells
表6实验结果表明，化合物和ADC对PSMA阳性细胞Vcap有一定的抑制增殖活性。The experimental results in Table 6 show that the compounds and ADC have certain inhibitory activity on Vcap of PSMA-positive cells.
FLOH1(PSMA基因名称)在不同细胞系中的表达水平参见The Human Protein Atlas(https://www.proteinatlas.org/)，数据显示在HL60细胞中不表达FLOH1；Vcap和22RV1表达FLOH1，详细结果见表7。The expression levels of FLOH1 (PSMA gene name) in different cell lines can be found in The Human Protein Atlas (https://www.proteinatlas.org/). The data shows that FLOH1 is not expressed in HL60 cells; Vcap and 22RV1 express FLOH1. Detailed results See Table 7.
表7 PSMA在不同细胞系中的表达水平
Table 7 Expression levels of PSMA in different cell lines
实施例6.6 ADC-002对22RV1移植瘤模型体内活性测试Example 6.6 In vivo activity test of ADC-002 on 22RV1 xenograft tumor model
1.实验细胞和实验动物1. Experimental cells and experimental animals
实验细胞:22RV1细胞，购自中国科学院细胞库； Experimental cells: 22RV1 cells, purchased from the Cell Bank of the Chinese Academy of Sciences;
实验动物:NOD/SCID小鼠，雄性，4-6周龄，购自维通利华实验动物有限公司。Experimental animals: NOD/SCID mice, male, 4-6 weeks old, purchased from Vitong Lever Experimental Animal Co., Ltd.
2.实验方案2. Experimental plan
2.1.细胞处理2.1. Cell processing
22RV1细胞，培养在含10％胎牛血清的RPMI-1640培养液中，约3-4天传代，细胞生长至足够数量后，用预冷的PBS与Matrigel(1:1)混合液调整细胞浓度至约2.5×107个/mL。22RV1 cells are cultured in RPMI-1640 culture medium containing 10% fetal calf serum and passaged in about 3-4 days. After the cells grow to a sufficient number, adjust the cell concentration with a mixture of pre-cooled PBS and Matrigel (1:1). to approximately 2.5×10 7 cells/mL.
2.2.肿瘤细胞移植2.2.Tumor cell transplantation
NOD/SCID小鼠实验室环境适应3-5天，于右前背部皮下接种22RV1细胞，接种细胞量为5×106cells/只，接种体积为0.2ml(含50％Matrigel)，待肿瘤生长至100～150mm3左右时，随机分组，进行实验。NOD/SCID mice were adapted to the laboratory environment for 3-5 days. 22RV1 cells were subcutaneously inoculated on the right front back at a volume of 5×10 6 cells/mouse and an inoculation volume of 0.2 ml (containing 50% Matrigel). Wait until the tumor grows to When the diameter is about 100~150mm 3 , they are randomly divided into groups and experiments are carried out.
2.3.动物给药与检测2.3. Animal administration and testing
表8入组的荷瘤裸鼠，按以下方案给药
The tumor-bearing nude mice enrolled in Table 8 were administered according to the following regimen.
Vehicle表示溶媒替代药物作为对照组。Vehicle indicates that the vehicle replaced the drug as a control group.
2.4.肿瘤体积及体重测定:2.4. Tumor volume and body weight determination:
每周测量2次瘤体积及体重，计算抑瘤率(TGI)(％)，计算公式为:TGI％＝(1-T/C)*100％(T和C分别为治疗组和对照组的相对肿瘤体积(V)。肿瘤体积(V)计算公式为:V＝0.5a*b2其中a和b分别表示肿瘤长径、短径。Measure the tumor volume and weight twice a week, and calculate the tumor inhibition rate (TGI) (%). The calculation formula is: TGI% = (1-T/C) * 100% (T and C are the values of the treatment group and the control group, respectively. Relative tumor volume (V). The calculation formula of tumor volume (V) is: V=0.5a*b 2 , where a and b represent the long diameter and short diameter of the tumor respectively.
3.实验结果3.Experimental results
具体结果见表9。ADC-002对22RV1的移植瘤模型，有较为显著的抗肿瘤生长的作用。至给药结束时(D21)，所有受试剂量组给药期间体重均轻微降低，无明显药物毒性。 The specific results are shown in Table 9. ADC-002 has a significant anti-tumor growth effect on the 22RV1 transplanted tumor model. By the end of the administration (D21), the body weight of all tested dose groups decreased slightly during the administration, and there was no obvious drug toxicity.
表9.ADC-002在第21天时对移植瘤模型22RV1的抑制作用
Table 9. Inhibitory effect of ADC-002 on transplanted tumor model 22RV1 on day 21
本领域技术人员将了解，上文描述本质上为示例性及说明性的，且欲说明本发明及其优选实施方案。通过常规实验，技术人员将了解可作出明显修正及变化而不悖离本发明的精神。在随附申请专利范围内的所有此类修正欲包括于其中。因此，本发明意欲并非由上述描述而是由以下权利要求范围及其等效物定义。Those skilled in the art will appreciate that the foregoing description is exemplary and explanatory in nature and is intended to describe the invention and its preferred embodiments. Through routine experimentation, the skilled artisan will realize that obvious modifications and changes may be made without departing from the spirit of the invention. All such modifications within the scope of the appended patent application are intended to be included therein. It is therefore intended that the invention be defined not by the above description but by the scope of the following claims and their equivalents.
本说明书中所引用的所有公开出版物以引用方式并入本文中。 All publications cited in this specification are hereby incorporated by reference.

Claims (18)

  1. 式I所示的抗体药物偶联物，
    Antibody drug conjugate represented by formula I,
    或所述抗体药物偶联物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物，or a stereoisomer of the antibody drug conjugate, a prodrug thereof, a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable solvate thereof,
    其中，in,
    Tb为抗体或其抗原结合片段；Tb is an antibody or its antigen-binding fragment;
    S为Tb上的硫原子；S is the sulfur atom on Tb;
    L为连接体，其共价结合Tb和D；L is the linker, which covalently binds Tb and D;
    D为Myc蛋白降解剂片段；D is the Myc protein degrader fragment;
    q为选自1-20的整数。q is an integer selected from 1-20.
  2. 式II所示的抗体药物偶联物，
    Antibody drug conjugate represented by formula II,
    或所述抗体药物偶联物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物，or a stereoisomer of the antibody drug conjugate, a prodrug thereof, a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable solvate thereof,
    其中，in,
    Tb为抗体或其抗原结合片段；Tb is an antibody or its antigen-binding fragment;
    S为Tb上的硫原子；S is the sulfur atom on Tb;
    L1为接头单元，其共价结合抗体或其抗原结合片段(Tb)与L2；L 1 is a linker unit, which covalently binds the antibody or its antigen-binding fragment (Tb) to L 2 ;
    L2为连接单元，其共价结合接头单元(L1)与L3；L 2 is a connecting unit, which covalently binds the linker unit (L 1 ) and L 3 ;
    L3选自氨基酸残基或由2-10个氨基酸残基组成的短肽；所述氨基酸残基选自天然氨基酸残基、非天然氨基酸残基或AA1所示氨基酸残基或其立体异构体；L 3 is selected from amino acid residues or short peptides composed of 2-10 amino acid residues; said amino acid residues are selected from natural amino acid residues, non-natural amino acid residues or amino acid residues shown in AA 1 or their stereoisomers conformation;
    AA1所示氨基酸残基的结构如下所示：
    The structure of the amino acid residue shown in AA 1 is as follows:
    Rx、Ry任一个为氢，另一个选自或者，Rx与Ry和与它们共同相连的碳原子一起，形成4-10元杂环，所述4-10元杂环任选地被一个或多个R0所取代；Either R x or R y is hydrogen, and the other is selected from Alternatively, R x and R y and the carbon atoms to which they are commonly connected form a 4-10 membered heterocyclic ring, which is optionally substituted by one or more R 0 ;
    Rx1、Ry1各自独立地选自氢、C1-6烷基；或者，Rx1与Ry1和与它们共同相连的氮原子一起，形成4-10元杂环，所述4-10元杂环任选地被一个或多个R0’所取代；R x1 and R y1 are each independently selected from hydrogen and C 1-6 alkyl; alternatively, R x1 and R y1 and the nitrogen atoms commonly connected to them form a 4-10 membered heterocyclic ring, and the 4-10 membered heterocycle The heterocycle is optionally substituted with one or more R 0' ;
    R0、R0’各自独立地选自C1-6烷基、C3-6环烷基、-NRx2Ry2和任选被C1-6烷基取代的4-10元杂环基；R 0 and R 0' are each independently selected from C 1-6 alkyl, C 3-6 cycloalkyl, -NR x2 R y2 and 4-10 membered heterocyclyl optionally substituted by C 1-6 alkyl. ;
    Rx2、Ry2各自独立地选自氢和C1-6烷基；R x2 and R y2 are each independently selected from hydrogen and C 1-6 alkyl;
    L4为化学键或间隔单元，共价结合D与L3；L 4 is a chemical bond or spacer unit, covalently combining D and L 3 ;
    D为Myc蛋白降解剂片段；D is the Myc protein degrader fragment;
    q为选自1-20的整数。q is an integer selected from 1-20.
  3. 如权利要求1或2所述的抗体药物偶联物或所述抗体药物偶联物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物，其特征在于：The antibody drug conjugate of claim 1 or 2 or the stereoisomer of the antibody drug conjugate, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate, Its characteristics are:
    (1)所述Tb为抗PSMA抗体或其抗原结合片段；(1) The Tb is an anti-PSMA antibody or an antigen-binding fragment thereof;
    (2)所述q选自1-12的整数；优选地，q选自1、2、3、4、5、6、7、8、9和10；优选地，所述q选自2、4、6、8；和/或；(2) The q is selected from an integer of 1-12; preferably, the q is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10; preferably, the q is selected from the group consisting of 2, 4, 6, 8; and/or;
    (3)所述L1选自1位通过S原子与Tb相连，2位与L2相连；和/或；(3) The L 1 is selected from Position 1 is connected to Tb through S atom, position 2 is connected to L 2 ; and/or;
    (4)L2选自1位与L1相连，2位与L3相连，W选自-CH2-、-OCH2CH2-，W’选自-CRx3Ry3-和-NRx3-，Rx3、Ry3各自独立地选自氢、C1-4烷基、C2-4烯基、C2-4炔基、C3-6环烷基、3-6元杂环基，或者，Rx3与Ry3和与它们共同 相连的碳原子一起，形成3-6元碳环或3-6元杂环，p为选自0-10之间的任意整数；和/或；(4)L 2 is selected from Bit 1 is connected to L 1 , bit 2 is connected to L 3 , W is selected from -CH 2 -, -OCH 2 CH 2 -, W' is selected from -CR x3 R y3 - and -NR x3 -, R x3 , R y3 Each is independently selected from hydrogen, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 3-6 cycloalkyl, 3-6 membered heterocyclyl, or R x3 and R y3 and common with them The connected carbon atoms together form a 3-6 membered carbocyclic ring or a 3-6 membered heterocyclic ring, and p is any integer selected from 0 to 10; and/or;
    (5)所述L3选自Val-Cit、Val-Ala、Phe-Lys、Val-Lys、Val-AA1、Ala-Ala-Ala、Ala-Ala-Asn、Val-AA1-Gly、Gly-Gly-Phe-Gly；优选地，L3选自Val-AA1-Gly；和/或；(5) The L 3 is selected from Val-Cit, Val-Ala, Phe-Lys, Val-Lys, Val-AA 1 , Ala-Ala-Ala, Ala-Ala-Asn, Val-AA 1 -Gly, Gly -Gly-Phe-Gly; Preferably, L 3 is selected from Val-AA 1 -Gly; and/or;
    (6)所述L4选自化学键、1位与L3相连，2位与D相连；和/或；(6) The L 4 is selected from chemical bonds, Bit 1 is connected to L 3 , bit 2 is connected to D; and/or;
    (7)所述D为式III所示的结构单元，D通过其含有的氧原子、氮原子与L4相连；
    (7) The D is a structural unit represented by formula III, and D is connected to L 4 through the oxygen atoms and nitrogen atoms it contains;
    其中，in,
    R1选自：Ra(CH2)a-、Ra(CH2)aC(＝O)-、Ra(CH2)aNHC(＝O)-、Ra(CH2)aOC(＝O)-或Ra(CH2)aS(＝O)2-，其中“CH2”任选被一个或多个R9取代，或者任选被-CH2CH2-取代，其中“NH”任选被R14取代；R 1 is selected from: R a (CH 2 ) a -, R a (CH 2 ) a C (=O)-, R a (CH 2 ) a NHC (= O) -, R a (CH 2 ) a OC (=O)- or R a (CH 2 ) a S(=O) 2 -, where "CH 2 " is optionally substituted by one or more R 9 , or optionally substituted by -CH 2 CH 2 -, where "NH" is optionally substituted by R 14 ;
    Q选自：-NR2-、-O-、 Q is selected from: -NR 2 -, -O-,
    R2选自：氢、Rb-、RbC(＝O)-、RbS(＝O)2-、R14；R 2 is selected from: hydrogen, R b -, R b C(=O)-, R b S(=O) 2 -, R 14 ;
    环A表示至少含有一个N原子作为杂原子的杂环烷基，其通过N原子与T相连，环A任选被一个或多个选自R9的基团取代；Ring A represents a heterocycloalkyl group containing at least one N atom as a heteroatom, which is connected to T through an N atom. Ring A is optionally substituted by one or more groups selected from R 9 ;
    Ra、Rb选自：C1-C8烷基、C3-C10环烷基、C3-C10桥环基、-NR11R12、C3-C10任含O、S、SO2、N或NHC(＝O)R22的杂环基、芳基、杂芳基、稠合芳基环烷基、稠合芳基杂环基、稠合杂芳基环烷基、稠合杂芳基杂环基、芳基-烷基、杂芳基-烷基、芳基-环烷基、芳基-杂环基、环烷基-杂环基、杂环基-杂环基，所述C1-C8烷基中的任意两个C-C之间可以插入选自-O-、-S-、-C(＝O)-、-S(＝O)-、-S(＝O)2-、 -NH-、-NHC(＝O)-、-NHC(＝O)NH-、-NHS(＝O)2-中的一种的基团，Ra任选被一个或多个R9取代，Rb任选被一个或多个R10取代；R a and R b are selected from: C 1 -C 8 alkyl, C 3 -C 10 cycloalkyl, C 3 -C 10 bridged cyclic group, -NR 11 R 12 , C 3 -C 10 optionally containing O and S , SO 2 , N or NHC(=O)R 22 heterocyclyl, aryl, heteroaryl, fused arylcycloalkyl, fused arylheterocyclyl, fused heteroarylcycloalkyl, Fused heteroarylheterocyclyl, aryl-alkyl, heteroaryl-alkyl, aryl-cycloalkyl, aryl-heterocyclyl, cycloalkyl-heterocyclyl, heterocyclyl-heterocycle group, any two CCs in the C 1 -C 8 alkyl group can be inserted between any two CCs selected from -O-, -S-, -C(=O)-, -S(=O)-, -S( ＝O) 2 -, A group of one of -NH-, -NHC(=O)-, -NHC(=O)NH-, -NHS(=O) 2 -, R a is optionally substituted by one or more R 9 , R b is optionally substituted by one or more R 10 ;
    T、U、Z分别独立地选自：化学键、羰基、C1-C6亚烷基、C3-C10亚环烷基、亚芳基、亚杂芳基、亚杂环基，所述亚烷基、亚环烷基、亚芳基、亚杂芳基、亚杂环基任选被一个或多个R9取代；T, U, and Z are each independently selected from: chemical bond, carbonyl group, C 1 -C 6 alkylene group, C 3 -C 10 cycloalkylene group, arylene group, heteroarylene group, heterocyclylene group, the above Alkylene, cycloalkylene, arylene, heteroarylene, heterocyclylene are optionally substituted by one or more R 9 ;
    Y选自：化学键、-C(＝O)NH(CH2)b-、-NHC(＝O)(CH2)b-、-O(CH2)b-、-NR2(CH2)b-，其中“CH2”任选被一个或多个R9取代，或者任选被-CH2CH2-取代，其中“NH”任选被R14取代；Y is selected from: chemical bond, -C(=O)NH(CH 2 ) b -, -NHC(=O)(CH 2 ) b -, -O(CH 2 ) b -, -NR 2 (CH 2 ) b -, wherein "CH 2 " is optionally substituted by one or more R 9 , or optionally substituted by -CH 2 CH 2 -, wherein "NH" is optionally substituted by R 14 ;
    X选自：-(CR7R8)o-、-C(＝O)-；X is selected from: -(CR 7 R 8 ) o -, -C(=O)-;
    R3-R5、R7-R9分别独立地选自：氢、R13、卤素、氰基、氨基、硝基、-R21C(＝O)R22、-R21NHC(＝O)R22、-R21C(＝O)NHR22、-R21C(＝O)OR22、-R21OC(＝O)R22、-R21S(＝O)2R22、-R21S(＝O)2NHR22、C1-C6烷基、C1-C6烷氧基、C1-C6烷硫基、C1-C6烷胺基、二(C1-C6烷基)胺基、C3-C8环烷基、C2-C8烯基、C2-C8炔基、芳基、含有1-3个杂原子的5-6元杂芳基或含有1-3个杂原子的3-10元杂环基，所述烷基、烷氧基、烷胺基、烷硫基、环烷基、芳基、杂芳基、杂环基任选被选自卤素、氰基、C1-C3烷基或C1-C3烷氧基的基团取代；当R3-R5、R7-R9为多个的情况下，任意相邻两个可结合形成环；R 3 -R 5 and R 7 -R 9 are each independently selected from: hydrogen, R 13 , halogen, cyano, amino, nitro, -R 21 C(=O)R 22 , -R 21 NHC(=O )R 22 , -R 21 C(=O)NHR 22 , -R 21 C(=O)OR 22 , -R 21 OC(=O)R 22 , -R 21 S(=O) 2 R 22 , - R 21 S(=O) 2 NHR 22 , C 1 -C 6 alkyl group, C 1 -C 6 alkoxy group, C 1 -C 6 alkylthio group, C 1 -C 6 alkylamino group, di(C 1 -C 6 alkyl)amine group, C 3 -C 8 cycloalkyl group, C 2 -C 8 alkenyl group, C 2 -C 8 alkynyl group, aryl group, 5-6 membered hetero group containing 1-3 heteroatoms Aryl or 3-10 membered heterocyclyl containing 1-3 heteroatoms, the alkyl, alkoxy, alkylamino, alkylthio, cycloalkyl, aryl, heteroaryl, heterocyclyl Optionally substituted with a group selected from halogen, cyano, C 1 -C 3 alkyl or C 1 -C 3 alkoxy; when R 3 -R 5 and R 7 -R 9 are multiple, Any two adjacent ones can be combined to form a ring;
    R10选自：氢、卤素、氰基、氨基、硝基、-R21C(＝O)R22、-R21NHC(＝O)R22、-R21C(＝O)NHR22、-R21C(＝O)OR22、-R21OC(＝O)R22、-R21S(＝O)2R22、-R21S(＝O)2NHR22、C1-C6烷基、C1-C6烷氧基、C1-C6烷硫基、C1-C6烷胺基、二(C1-C6烷基)胺基、C3-C8环烷基、C2-C8烯基、C2-C8炔基、芳基、含有1-3个杂原子的5-6元杂芳基或含有1-3个杂原子的3-10元杂环基，所述烷基、烷氧基、烷胺基、烷硫基、环烷基、芳基、杂芳基、杂环基任选被选自卤素、氰基、C1-C3烷基或C1-C3烷氧基的基团取代；当R10为多个的情况下，任意相邻两个可结合形成环；R 10 is selected from: hydrogen, halogen, cyano, amino, nitro, -R 21 C(=O)R 22 , -R 21 NHC(=O)R 22 , -R 21 C(=O)NHR 22 , -R 21 C(=O)OR 22 , -R 21 OC(=O)R 22 , -R 21 S(=O) 2 R 22 , -R 21 S(=O) 2 NHR 22 , C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkylthio, C 1 -C 6 alkylamino, di(C 1 -C 6 alkyl)amine, C 3 -C 8 ring Alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, aryl, 5-6 membered heteroaryl containing 1-3 heteroatoms or 3-10 membered containing 1-3 heteroatoms Heterocyclyl group, the alkyl group, alkoxy group, alkylamino group, alkylthio group, cycloalkyl group, aryl group, heteroaryl group, heterocyclyl group are optionally selected from halogen, cyano group, C 1 -C 3 Alkyl or C 1 -C 3 alkoxy group substitution; when there are multiple R 10s , any two adjacent ones can be combined to form a ring;
    R13选自：羟基、Rn、-ORn、-C(＝O)Rn、-C(＝O)ORn、-OC(＝O)Rn、-NRmRn、-C(＝O)NRmRn、-NRm-C(＝O)Rn、-NRn-C(＝O)Rm、-S(＝O)2Rn、-S(＝O)2NRmRn、-NRm-S(＝O)2Rn、-NRn-S(＝O)2Rm；R 13 is selected from: hydroxyl, R n , -OR n , -C(=O)R n , -C(=O)OR n , -OC(=O)R n , -NR m R n , -C( =O)NR m R n , -NR m -C(=O)R n , -NR n -C(=O)R m , -S(=O) 2 R n , -S(=O) 2 NR m R n , -NR m -S(=O) 2 R n , -NR n -S(=O) 2 R m ;
    R14选自：Rn、-C(＝O)Rn、-C(＝O)ORn、-C(＝O)NRmRn、-S(＝O)2Rn、-S(＝O)2NRmRn； R 14 is selected from: R n , -C(=O)R n , -C(=O)OR n , -C(=O)NR m R n , -S(=O) 2 R n , -S( ＝O) 2 NR m R n ;
    Rn选自：被羟基取代的C1-C8烷基，所述C1-C8烷基中的任意两个C-C之间可以插入选自-O-、-S-、-C(＝O)-、-C(＝O)O-、-OC(＝O)O-、-S(＝O)-、-S(＝O)2-、-NH-、-NHC(＝O)-、-NHC(＝O)NH-、-NHS(＝O)2-中的一种的基团，所述C1-C8烷基任选被1-3个分别选自卤素、氰基、氨基、硝基或C1-C3烷氧基的基团取代；R n is selected from: C 1 -C 8 alkyl groups substituted by hydroxyl groups, and any two CCs in the C 1 -C 8 alkyl groups can be inserted between -O-, -S-, -C(= O)-, -C(=O)O-, -OC(=O)O-, -S(=O)-, -S(=O) 2 -, -NH-, -NHC(=O)- , -NHC(=O)NH-, -NHS(=O) 2 -, the C 1 -C 8 alkyl group is optionally selected from 1-3 halogen, cyano, Substitution of amino, nitro or C 1 -C 3 alkoxy groups;
    Rm独立地选自：氢、C1-C8烷基，所述C1-C8烷基中的任意两个C-C之间可以插入选自-O-、-S-、-C(＝O)-、-C(＝O)O-、-OC(＝O)O-、-S(＝O)-、-S(＝O)2-、-NH-、-NHC(＝O)-、-NHC(＝O)NH-、-NHS(＝O)2-中的一种的基团，所述C1-C8烷基任选被1-3个分别选自卤素、羟基、氰基、氨基、硝基或C1-C3烷氧基的基团取代；R6选自：氢、C1-C6烷基、C3-C8环烷基、C2-C8烯基、C2-C8炔基、芳基、含有1-3个杂原子的5-6元杂芳基或含有1-3个杂原子的3-10元杂环基，所述烷基、环烷基、芳基、杂芳基、杂环基任选被1-3个分别选自卤素、羟基、氰基、氨基、硝基、-C(＝O)OR23、-OC(＝O)R23、-NHC(＝O)R23、-C(＝O)NHR23、C1-C3烷基、C1-C3烷氧基或-OP(＝O)(OM)2的基团取代；R m is independently selected from: hydrogen, C 1 -C 8 alkyl group, and any two CC in the C 1 -C 8 alkyl group can be inserted between any two CCs selected from -O-, -S-, -C(= O)-, -C(=O)O-, -OC(=O)O-, -S(=O)-, -S(=O) 2 -, -NH-, -NHC(=O)- , -NHC(=O)NH-, -NHS(=O) 2 -, the C 1 -C 8 alkyl group is optionally selected from 1-3 halogen, hydroxyl, cyanide, etc. Substituted with a base, amino, nitro or C 1 -C 3 alkoxy group; R 6 is selected from: hydrogen, C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl, C 2 -C 8 alkene base, C 2 -C 8 alkynyl group, aryl group, 5-6 membered heteroaryl group containing 1-3 heteroatoms or 3-10 membered heterocyclyl group containing 1-3 heteroatoms, the alkyl group, Cycloalkyl, aryl, heteroaryl and heterocyclic groups are optionally selected from 1 to 3 halogen, hydroxyl, cyano, amino, nitro, -C(=O)OR 23 , -OC(=O )R 23 , -NHC(=O)R 23 , -C(=O)NHR 23 , C 1 -C 3 alkyl, C 1 - C 3 alkoxy or -OP(=O)(OM) 2 group substitution;
    M独立地选自：氢、C1-C4烷基；M is independently selected from: hydrogen, C 1 -C 4 alkyl;
    R11、R12分别独立地选自：氢、C1-C4烷基、芳基、芳基-烷基；R 11 and R 12 are each independently selected from: hydrogen, C 1 -C 4 alkyl, aryl, aryl-alkyl;
    R21选自：化学键、C1-C4亚烷基；R 21 is selected from: chemical bond, C 1 -C 4 alkylene;
    R22、R23分别独立地选自：氢、C1-C4烷基、芳基、芳基-烷基，所述C1-C4烷基、芳基、芳基-烷基任选被选自卤素、羟基、氨基的基团取代；R 22 and R 23 are each independently selected from: hydrogen, C 1 -C 4 alkyl, aryl, aryl-alkyl, the C 1 -C 4 alkyl, aryl, aryl-alkyl is optional Substituted with a group selected from halogen, hydroxyl, and amino;
    a选自：0、1、2、3、4或5；a is selected from: 0, 1, 2, 3, 4 or 5;
    b选自：0、1、2、3、4或5；b is selected from: 0, 1, 2, 3, 4 or 5;
    n选自：0、1、2或3；n is selected from: 0, 1, 2 or 3;
    m选自：0、1、2、3或4；m is selected from: 0, 1, 2, 3 or 4;
    o选自：1或2；oSelected from: 1 or 2;
    其中，式III所示的结构单元中至少包含一个选自R13或R14的基团。Wherein, the structural unit represented by formula III contains at least one group selected from R 13 or R 14 .
  4. 如权利要求1或2所述的抗体药物偶联物或所述抗体药物偶联物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物，其特征在于：The antibody drug conjugate of claim 1 or 2 or the stereoisomer of the antibody drug conjugate, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate, Its characteristics are:
    (1)所述Tb为抗PSMA抗体或其抗原结合片段； (1) The Tb is an anti-PSMA antibody or an antigen-binding fragment thereof;
    (2)所述q选自1-12的整数；优选地，q选自1、2、3、4、5、6、7、8、9和10；优选地，所述q选自2、4、6、8；和/或；(2) The q is selected from an integer of 1-12; preferably, the q is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10; preferably, the q is selected from the group consisting of 2, 4, 6, 8; and/or;
    (3)所述L1选自1位通过S原子与Tb相连，2位与L2相连；和/或；(3) The L 1 is selected from Position 1 is connected to Tb through S atom, position 2 is connected to L 2 ; and/or;
    (4)L2选自1位与L1相连，2位与L3相连，W选自-CH2-、-OCH2CH2-，W’选自-CRx3Ry3-和-NRx3-，Rx3、Ry3各自独立地选自氢、C1-4烷基、C2-4烯基、C2-4炔基、C3-6环烷基、3-6元杂环基，或者，Rx3与Ry3和与它们共同相连的碳原子一起，形成3-6元碳环或3-6元杂环，p为选自0-10之间的任意整数；和/或；(4)L 2 is selected from Bit 1 is connected to L 1 , bit 2 is connected to L 3 , W is selected from -CH 2 -, -OCH 2 CH 2 -, W' is selected from -CR x3 R y3 - and -NR x3 -, R x3 , R y3 Each is independently selected from hydrogen, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 3-6 cycloalkyl, 3-6 membered heterocyclyl, or R x3 and R y3 and the carbon atoms connected to them together form a 3-6-membered carbocyclic ring or a 3-6-membered heterocyclic ring, p is any integer selected from 0-10; and/or;
    (5)所述L3选自Val-Cit、Val-Ala、Phe-Lys、Val-Lys、Val-AA1、Ala-Ala-Ala、Ala-Ala-Asn、Val-AA1-Gly、Gly-Gly-Phe-Gly；优选地，L3选自Val-AA1-Gly；和/或；(5) The L 3 is selected from Val-Cit, Val-Ala, Phe-Lys, Val-Lys, Val-AA 1 , Ala-Ala-Ala, Ala-Ala-Asn, Val-AA 1 -Gly, Gly -Gly-Phe-Gly; Preferably, L 3 is selected from Val-AA 1 -Gly; and/or;
    (6)所述L4选自化学键、1位与L3相连，2位与D相连；和/或；(6) The L 4 is selected from chemical bonds, Bit 1 is connected to L 3 , bit 2 is connected to D; and/or;
    (7)所述D为式III所示的结构单元，D通过其含有的氧原子、氮原子或硫原子与L4相连；
    (7) The D is a structural unit represented by formula III, and D is connected to L 4 through the oxygen atom, nitrogen atom or sulfur atom it contains;
    其中，in,
    R1选自：Ra(CH2)a-、Ra(CH2)aC(＝O)-、Ra(CH2)aNHC(＝O)-、Ra(CH2)aOC(＝O)-或Ra(CH2)aS(＝O)2-，其中“CH2”任选被一个或多个R9取代，或者任选被-CH2CH2-取代，其中“NH”任选被R14取代； R 1 is selected from: R a (CH 2 ) a -, R a (CH 2 ) a C (=O)-, R a (CH 2 ) a NHC (= O) -, R a (CH 2 ) a OC (=O)- or R a (CH 2 ) a S(=O) 2 -, where "CH 2 " is optionally substituted by one or more R 9 , or optionally substituted by -CH 2 CH 2 -, where "NH" is optionally substituted by R 14 ;
    Q选自：-NR2-、-O-、 Q is selected from: -NR 2 -, -O-,
    R2选自：氢、Rb-、RbC(＝O)-、RbS(＝O)2-、R14；R 2 is selected from: hydrogen, R b -, R b C(=O)-, R b S(=O) 2 -, R 14 ;
    环A表示至少含有一个N原子作为杂原子的杂环烷基，其通过N原子与T或R1相连，环A任选被一个或多个选自R9的基团取代；Ring A represents a heterocycloalkyl group containing at least one N atom as a heteroatom, which is connected to T or R 1 through an N atom. Ring A is optionally substituted by one or more groups selected from R 9 ;
    Ra、Rb选自：C1-C8烷基、C3-C10环烷基、C3-C10桥环基、-NR11R12、C3-C10任含O、S、SO2、N或NHC(＝O)R22的杂环基、芳基、杂芳基、稠合芳基环烷基、稠合芳基杂环基、稠合杂芳基环烷基、稠合杂芳基杂环基、芳基-芳基、芳基-杂芳基、杂芳基-芳基、杂芳基-杂芳基、芳基-烷基、杂芳基-烷基、芳基-环烷基、芳基-杂环基、环烷基-杂环基、杂环基-杂环基，所述C1-C8烷基中的任意两个C-C之间可以插入选自-O-、-S-、-C(＝O)-、-S(＝O)-、-S(＝O)2-、-NH-、-NHC(＝O)-、-NHC(＝O)NH-、-NHS(＝O)2-中的一种的基团，Ra任选被一个或多个R9取代，Rb任选被一个或多个R10取代；R a and R b are selected from: C 1 -C 8 alkyl, C 3 -C 10 cycloalkyl, C 3 -C 10 bridged cyclic group, -NR 11 R 12 , C 3 -C 10 optionally containing O and S , SO 2 , N or NHC(=O)R 22 heterocyclyl, aryl, heteroaryl, fused arylcycloalkyl, fused arylheterocyclyl, fused heteroarylcycloalkyl, Fused heteroarylheterocyclyl, aryl-aryl, aryl-heteroaryl, heteroaryl-aryl, heteroaryl-heteroaryl, aryl-alkyl, heteroaryl-alkyl, Aryl-cycloalkyl, aryl-heterocyclyl, cycloalkyl-heterocyclyl, heterocyclyl-heterocyclyl, optional carbon atoms can be inserted between any two CCs in the C 1 -C 8 alkyl group. From -O-, -S-, -C(=O)-, -S(=O)-, -S(=O) 2 -, -NH-, -NHC(=O)-, -NHC(= O) A group of one of NH- and -NHS(=O) 2 -, R a is optionally substituted by one or more R 9 , R b is optionally substituted by one or more R 10 ;
    T、U、Z分别独立地选自：化学键、N、O、羰基、C1-C6亚烷基、C3-C10亚环烷基、亚芳基、亚杂芳基、亚杂环基，所述亚烷基、亚环烷基、亚芳基、亚杂芳基、亚杂环基任选被一个或多个R9取代；T, U, and Z are each independently selected from: chemical bond, N, O, carbonyl group, C 1 -C 6 alkylene group, C 3 -C 10 cycloalkylene group, arylene group, heteroarylene group, heterocyclic ring Base, the alkylene, cycloalkylene, arylene, heteroarylene, heterocyclylene is optionally substituted by one or more R 9 ;
    Y选自：化学键、-C(＝O)NH(CH2)b-、-NHC(＝O)(CH2)b-、-O(CH2)b-、-NR2(CH2)b-，其中“CH2”任选被一个或多个R9取代，或者任选被-CH2CH2-取代，其中“NH”任选被R14取代；Y is selected from: chemical bond, -C(=O)NH(CH 2 ) b -, -NHC(=O)(CH 2 ) b -, -O(CH 2 ) b -, -NR 2 (CH 2 ) b -, wherein "CH 2 " is optionally substituted by one or more R 9 , or optionally substituted by -CH 2 CH 2 -, wherein "NH" is optionally substituted by R 14 ;
    X选自：-(CR7R8)o-、-C(＝O)-；X is selected from: -(CR 7 R 8 ) o -, -C(=O)-;
    R3-R5、R7-R9分别独立地选自：氢、R13、卤素、氰基、氨基、羟基、巯基、硝基、-R21N(R22)R22、-R21C(＝O)R22、-R21NHC(＝O)R22、-R21C(＝O)NHR22、-R21C(＝O)OR22、-R21OC(＝O)R22、-R21S(＝O)2R22、-R21S(＝O)2NHR22、C1-C6烷基、C1-C6烷氧基、C1-C6烷硫基、C1-C6烷胺基、二(C1-C6烷基)胺基、C3-C8环烷基、C2-C8烯基、C2-C8炔基、芳基、含有1-3个杂原子的5-6元杂芳基或含有1-3个杂原子的3-10元杂环基，所述烷基、烷氧基、烷胺基、烷硫基、环烷基、芳基、杂芳基、杂环基任选被选自卤素、氰基、C1-C3烷基或C1-C3烷氧基的基团取代；当R3-R5、R7-R9为多个的情况下，任意相邻两个可结合形成环；R 3 -R 5 and R 7 -R 9 are each independently selected from: hydrogen, R 13 , halogen, cyano, amino, hydroxyl, mercapto, nitro, -R 21 N(R 22 )R 22, -R 21 C(=O)R 22 , -R 21 NHC(=O)R 22 , -R 21 C(=O)NHR 22 , -R 21 C(=O)OR 22 , -R 21 OC(=O)R 22 , -R 21 S(＝O) 2 R 22 , -R 21 S(＝O) 2 NHR 22 , C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkylthio base, C 1 -C 6 alkylamino group, di(C 1 -C 6 alkyl)amine group, C 3 -C 8 cycloalkyl group, C 2 -C 8 alkenyl group, C 2 -C 8 alkynyl group, aromatic group base, a 5-6-membered heteroaryl group containing 1-3 heteroatoms or a 3-10-membered heterocyclic group containing 1-3 heteroatoms, the alkyl group, alkoxy group, alkylamino group, alkylthio group , cycloalkyl, aryl, heteroaryl, heterocyclyl optionally substituted with a group selected from halogen, cyano, C 1 -C 3 alkyl or C 1 -C 3 alkoxy; when R 3 - When there are multiple R 5 , R 7 -R 9 , any two adjacent ones can be combined to form a ring;
    R10选自：氢、卤素、氰基、氨基、硝基、-R21C(＝O)R22、-R21NHC(＝O)R22、-R21C(＝O)NHR22、-R21C(＝O)OR22、-R21OC(＝O)R22、-R21S(＝O)2R22、 -R21S(＝O)2NHR22、C1-C6烷基、C1-C6烷氧基、C1-C6烷硫基、C1-C6烷胺基、二(C1-C6烷基)胺基、C3-C8环烷基、C2-C8烯基、C2-C8炔基、芳基、含有1-3个杂原子的5-6元杂芳基或含有1-3个杂原子的3-10元杂环基，所述烷基、烷氧基、烷胺基、烷硫基、环烷基、芳基、杂芳基、杂环基任选被选自卤素、氰基、C1-C3烷基或C1-C3烷氧基的基团取代；当R10为多个的情况下，任意相邻两个可结合形成环；R 10 is selected from: hydrogen, halogen, cyano, amino, nitro, -R 21 C(=O)R 22 , -R 21 NHC(=O)R 22 , -R 21 C(=O)NHR 22 , -R 21 C(＝O)OR 22 , -R 21 OC(＝O)R 22 , -R 21 S(＝O) 2 R 22 , -R 21 S(=O) 2 NHR 22 , C 1 -C 6 alkyl group, C 1 -C 6 alkoxy group, C 1 -C 6 alkylthio group, C 1 -C 6 alkylamino group, di(C 1 -C 6 alkyl)amine, C 3 -C 8 cycloalkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, aryl, 5-6 yuan containing 1-3 heteroatoms Heteroaryl or 3-10 membered heterocyclyl containing 1-3 heteroatoms, the alkyl, alkoxy, alkylamino, alkylthio, cycloalkyl, aryl, heteroaryl, heterocycle The group is optionally substituted with a group selected from halogen, cyano, C 1 -C 3 alkyl or C 1 -C 3 alkoxy; when there are multiple R 10s , any two adjacent ones can be combined to form ring;
    R13选自：羟基、巯基、氨基、Rn、-ORn、-C(＝O)Rn、-C(＝O)ORn、-OC(＝O)Rn、-NRmRn、-C(＝O)NRmRn、-NRm-C(＝O)Rn、-NRn-C(＝O)Rm、-S(＝O)2Rn、-S(＝O)2NRmRn、-NRm-S(＝O)2Rn、-NRn-S(＝O)2Rm；R 13 is selected from: hydroxyl, mercapto, amino, R n , -OR n , -C(=O)R n , -C(=O)OR n , -OC(=O)R n , -NR m R n , -C(=O)NR m R n , -NR m -C(=O)R n , -NR n -C(=O)R m , -S(=O) 2 R n , -S(= O) 2 NR m R n , -NR m -S(=O) 2 R n , -NR n -S(=O) 2 R m ;
    R14选自：Rn、-C(＝O)Rn、-C(＝O)ORn、-C(＝O)NRmRn、-S(＝O)2Rn、-S(＝O)2NRmRn；Rn选自：被羟基、巯基或氨基取代的C1-C8烷基，所述C1-C8烷基中的任意两个C-C之间可以插入选自-O-、-S-、-C(＝O)-、-C(＝O)O-、-OC(＝O)O-、-S(＝O)-、-S(＝O)2-、-NH-、-NHC(＝O)-、-NHC(＝O)NH-、-NHS(＝O)2-中的一种的基团，所述C1-C8烷基任选被1-3个分别选自卤素、氰基、氨基、硝基或C1-C3烷氧基的基团取代；Rm独立地选自：氢、C1-C8烷基，所述C1-C8烷基中的任意两个C-C之间可以插入选自-O-、-S-、-C(＝O)-、-C(＝O)O-、-OC(＝O)O-、-S(＝O)-、-S(＝O)2-、-NH-、-NHC(＝O)-、-NHC(＝O)NH-、-NHS(＝O)2-中的一种的基团，所述C1-C8烷基任选被1-3个分别选自卤素、羟基、氰基、氨基、硝基或C1-C3烷氧基的基团取代；R6选自：氢、C1-C6烷基、C3-C8环烷基、C2-C8烯基、C2-C8炔基、芳基、含有1-3个杂原子的5-6元杂芳基或含有1-3个杂原子的3-10元杂环基，所述烷基、环烷基、芳基、杂芳基、杂环基任选被1-3个分别选自卤素、羟基、氰基、氨基、硝基、-C(＝O)OR23、-OC(＝O)R23、-NHC(＝O)R23、-C(＝O)NHR23、C1-C3烷基、C1-C3烷氧基或-OP(＝O)(OM)2的基团取代；R 14 is selected from: R n , -C(=O)R n , -C(=O)OR n , -C(=O)NR m R n , -S(=O) 2 R n , -S( =O) 2 NR m R n ; R n is selected from: C 1 -C 8 alkyl groups substituted by hydroxyl, mercapto or amino groups, and optional C 1 -C 8 alkyl groups can be inserted between any two. From -O-, -S-, -C(=O)-, -C(=O)O-, -OC(=O)O-, -S(=O)-, -S(=O) 2 -, -NH-, -NHC(=O)-, -NHC(=O)NH-, -NHS(=O) 2 -, the C 1 -C 8 alkyl group is optional Substituted by 1-3 groups respectively selected from halogen, cyano, amino, nitro or C 1 -C 3 alkoxy; R m is independently selected from: hydrogen, C 1 -C 8 alkyl, the Between any two CCs in the C 1 -C 8 alkyl group, a member selected from -O-, -S-, -C(=O)-, -C(=O)O-, -OC(=O) can be inserted O-, -S(＝O)-, -S(＝O) 2 -, -NH-, -NHC(＝O)-, -NHC(＝O)NH-, -NHS(＝O) 2 -中A kind of group, the C 1 -C 8 alkyl group is optionally substituted by 1-3 groups respectively selected from halogen, hydroxyl, cyano, amino, nitro or C 1 -C 3 alkoxy group ; R 6 is selected from: hydrogen, C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, aryl, containing 1-3 hetero 5-6-membered heteroaryl group or 3-10-membered heterocyclyl group containing 1-3 heteroatoms, and the alkyl group, cycloalkyl group, aryl group, heteroaryl group and heterocyclyl group are optionally substituted by 1- 3 are respectively selected from halogen, hydroxyl, cyano, amino, nitro, -C(=O)OR 23 , -OC(=O)R 23 , -NHC(=O)R 23 , -C(=O) NHR 23 , C 1 -C 3 alkyl, C 1 - C 3 alkoxy or -OP(=O)(OM) 2 group substitution;
    M独立地选自：氢、C1-C4烷基；M is independently selected from: hydrogen, C 1 -C 4 alkyl;
    R11、R12分别独立地选自：氢、C1-C4烷基、芳基、芳基-烷基；R 11 and R 12 are each independently selected from: hydrogen, C 1 -C 4 alkyl, aryl, aryl-alkyl;
    R21选自：化学键、C1-C4亚烷基；R 21 is selected from: chemical bond, C 1 -C 4 alkylene;
    R22、R23分别独立地选自：氢、C1-C4烷基、芳基、芳基-烷基，所述C1-C4烷基、芳基、芳基-烷基任选被选自卤素、羟基、氨基的基团取代；R 22 and R 23 are each independently selected from: hydrogen, C 1 -C 4 alkyl, aryl, aryl-alkyl, the C 1 -C 4 alkyl, aryl, aryl-alkyl is optional Substituted with a group selected from halogen, hydroxyl, and amino;
    a选自：0、1、2、3、4或5；a is selected from: 0, 1, 2, 3, 4 or 5;
    b选自：0、1、2、3、4或5； b is selected from: 0, 1, 2, 3, 4 or 5;
    n选自：0、1、2或3；n is selected from: 0, 1, 2 or 3;
    m选自：0、1、2、3或4；m is selected from: 0, 1, 2, 3 or 4;
    o选自：1或2；oSelected from: 1 or 2;
    优选的，式III所示的结构单元中至少包含一个选自R13或R14的基团。Preferably, the structural unit represented by formula III contains at least one group selected from R 13 or R 14 .
  5. 如权利要求1-4任一项所述的抗体药物偶联物或所述抗体药物偶联物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物，其中The antibody drug conjugate according to any one of claims 1 to 4 or the stereoisomer of the antibody drug conjugate, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvent compound, among which
    R1选自：Ra-、RaCH2-、RaCH2CH2-、RaCH2C(＝O)-和RaCH(OH)CH2-；R 1 is selected from: R a -, R a CH 2 -, R a CH 2 CH 2 -, R a CH 2 C(=O)-, and R a CH(OH)CH 2 -;
    所述Ra选自以下基团：
    The R a is selected from the following groups:
    所述Ra进一步优选为以下基团：
    The Ra is further preferably the following group:
    Q选自：-NR2-和 Q is selected from: -NR 2 -and
    R2选自：氢、-C1-C4烷基、Rn、C(＝O)Rn、-C(＝O)ORn和-C(＝O)NRmRn；R 2 is selected from: hydrogen, -C 1 -C 4 alkyl, R n , C(=O)R n , -C(=O)OR n and -C(=O)NR m R n ;
    Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH和-CH2CH2CH2CH2OH；优选地，所述Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH、-CH2CH2CH2CH2OH、-CH2NH2、-CH2CH2NH2、-CH2CH2CH2NH2、-CH2CH2CH2CH2NH2、-CH2SH、-CH2CH2SH、-CH2CH2CH2SH、-CH2CH2CH2CH2SH；R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH and -CH 2 CH 2 CH 2 CH 2 OH; preferably, the R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 CH 2 SH;
    Rm为氢；R m is hydrogen;
    选自以下结构中的一种：
     Choose one of the following structures:
    且通过N原子与T或R1相连；and Connected to T or R 1 through N atom;
    -T-U-Z-一起形成选自以下的基团：-T-U-Z- together form a group selected from:
    和C1-4亚烷基(优选亚甲基)； and C 1-4 alkylene (preferably methylene);
    优选地，当Q为-NR2-时，-T-U-Z-不为C1-4亚烷基；Y选自-C(＝O)NHCH2-、-C(＝O)NH和-NHC(＝O)-；Preferably, when Q is -NR 2 -, -TUZ- is not C 1-4 alkylene; Y is selected from -C(=O)NHCH 2 -, -C(=O)NH and -NHC(= O)-;
    R3-R6分别独立地为氢；R 3 to R 6 are each independently hydrogen;
    X为CH2；X is CH 2 ;
    m和n为0。m and n are 0.
  6. 如权利要求1-5中任一项所述的抗体药物偶联物或所述抗体药物偶联物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物，其中R1选自：Ra-、RaCH2-、RaCH2CH2-、RaCH2C(＝O)-和RaCH(OH)CH2-；The antibody drug conjugate according to any one of claims 1 to 5 or the stereoisomer of the antibody drug conjugate, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable salt Solvates, wherein R 1 is selected from: R a -, R a CH 2 -, R a CH 2 CH 2 -, R a CH 2 C(=O)-, and R a CH(OH)CH 2 -;
    所述Ra选自以下基团：
    The R a is selected from the following groups:
    所述Ra进一步优选为以下基团：
    The Ra is further preferably the following group:
    Q选自：-NR2-和 Q is selected from: -NR 2 -and
    R2选自：-C1-C4烷基、-C(＝O)ORn和-C(＝O)NRmRn；R 2 is selected from: -C 1 -C 4 alkyl, -C(=O)OR n and -C(=O)NR m R n ;
    Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH、-CH2CH2CH2CH2OH、-CH2NH2、-CH2CH2NH2、-CH2CH2CH2NH2、-CH2CH2CH2CH2NH2、-CH2SH、-CH2CH2SH、-CH2CH2CH2SH、-CH2CH2CH2CH2SH；R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 CH 2 OH, -CH 2 NH 2 , -CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 , -CH 2 SH, -CH 2 CH 2 SH, -CH 2 CH 2 CH 2 SH, -CH 2 CH 2 CH 2 CH 2 SH;
    优选地，所述Rn选自：-CH2OH、-CH2CH2OH、-CH2CH2CH2OH和-CH2CH2CH2CH2OH；Preferably, the R n is selected from: -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH and -CH 2 CH 2 CH 2 CH 2 OH;
    Rm为氢；R m is hydrogen;
    选自以下结构中的一种：
     Choose one of the following structures:
    -T-U-Z-一起形成选自以下的基团：
    -TUZ- together form a group selected from:
    Y选自-C(＝O)NHCH2-和-NHC(＝O)-；Y is selected from -C(=O)NHCH 2 - and -NHC(=O)-;
    R3-R6分别独立地为氢；R 3 to R 6 are each independently hydrogen;
    X为CH2； X is CH 2 ;
    m和n为0。m and n are 0.
  7. 如权利要求3-6任一项所述的抗体药物偶联物或所述抗体药物偶联物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物，其中所述式III所示的结构单元为如下式III-A所示的结构单元
    The antibody drug conjugate according to any one of claims 3 to 6 or the stereoisomer of the antibody drug conjugate, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvent compound, wherein the structural unit represented by formula III is a structural unit represented by the following formula III-A
    其中R1、R3、R4、R5、R6、Q、T、U、Z、X、Y、m和n如权利要求3-5之一所定义，wherein R 1 , R 3 , R 4 , R 5 , R 6 , Q, T, U, Z, X, Y, m and n are as defined in one of claims 3 to 5,
    优选地所述式III所示的结构单元为如下所示结构


    Preferably, the structural unit represented by Formula III has the following structure:


  8. 如权利要求1-7任一项所述的抗体药物偶联物或所述抗体药物偶联物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物，其中PSMA抗体或其抗原结合片段为含有如下CDR的抗体或其抗原结合片段：SEQ ID No.7所示的HCDR1，SEQ ID No.8所示的HCDR2，SEQ ID No.9所示的HCDR3，SEQ ID No.10所示的LCDR1，SEQ ID No.11所示的LCDR2，SEQ ID No.12所示的LCDR3；The antibody drug conjugate according to any one of claims 1 to 7 or the stereoisomer of the antibody drug conjugate, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvent A compound, wherein the PSMA antibody or its antigen-binding fragment is an antibody or its antigen-binding fragment containing the following CDRs: HCDR1 shown in SEQ ID No.7, HCDR2 shown in SEQ ID No.8, SEQ ID No.9 HCDR3, LCDR1 shown in SEQ ID No.10, LCDR2 shown in SEQ ID No.11, LCDR3 shown in SEQ ID No.12;
    优选地，所述PSMA抗体或其抗原结合片段含有SEQ ID No.1所示的VH和SEQ ID No.3所示的VL；Preferably, the PSMA antibody or antigen-binding fragment thereof contains the VH shown in SEQ ID No. 1 and the VL shown in SEQ ID No. 3;
    优选地，所述PSMA抗体或其抗原结合片段含有：SEQ ID No.1所示的VH和SEQ ID No.2所示的CH，和SEQ ID No.3所示的VL和SEQ ID No.4所示的CL；Preferably, the PSMA antibody or antigen-binding fragment thereof contains: VH shown in SEQ ID No. 1 and CH shown in SEQ ID No. 2, and VL shown in SEQ ID No. 3 and SEQ ID No. 4 CL shown;
    最优选地，所述PSMA抗体或其抗原结合片段含有：SEQ ID No.5所示的重链和SEQ ID No.6所示的轻链。Most preferably, the PSMA antibody or antigen-binding fragment thereof contains: the heavy chain shown in SEQ ID No. 5 and the light chain shown in SEQ ID No. 6.
  9. 如权利要求1-8任一项所述的抗体药物偶联物或所述抗体药物偶联物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物，其特征在于，结构如式IV所示，
    The antibody drug conjugate according to any one of claims 1 to 8 or the stereoisomer of the antibody drug conjugate, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvent The compound is characterized in that the structure is shown in formula IV,
    其中，Tb、S、L1、L2、Rx1、Ry1、D和q的定义如权利要求1-8任一项所定义。Among them, the definitions of Tb, S, L 1 , L 2 , R x1 , R y1 , D and q are as defined in any one of claims 1 to 8.
  10. 如权利要求1-9任一项所述的抗体药物偶联物，其特征在于，所述的抗体药物偶联物选自：








    The antibody drug conjugate according to any one of claims 1 to 9, characterized in that the antibody drug conjugate is selected from:








    其中，q选自1-10的整数，例如2-8的整数。Wherein, q is selected from an integer of 1-10, such as an integer of 2-8.
  11. 权利要求1-10任一项所述的抗体药物偶联物或所述抗体药物偶联物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物，特征在于，其具有选自1.0-10.0内的任意数值的平均DAR；优选地，所述的抗体药物偶联物，特征在于其具有选自2.0-8.0内的任意数值的平均DAR。The antibody drug conjugate of any one of claims 1 to 10 or the stereoisomer of the antibody drug conjugate, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate The object is characterized by having an average DAR of any value selected from 1.0-10.0; preferably, the antibody drug conjugate is characterized by having an average DAR of any value selected from 2.0-8.0.
  12. 式V所示的药物连接体偶联物，
    L′-D
    式V    Drug linker conjugate represented by formula V,
    L′-D
    Formula V
    或所述药物连接体偶联物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物；Or the stereoisomer of the drug linker conjugate, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate;
    其中，L’为连接体前体，D的定义如权利要求1-11任一项所定义。 Wherein, L' is a linker precursor, and D is as defined in any one of claims 1-11.
  13. 式VI所示的药物连接体偶联物或所述药物连接体偶联物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物，
    Lg-L1-L2-L3-L4-D
    式VI    The drug linker conjugate represented by Formula VI or the stereoisomer of the drug linker conjugate, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate,
    Lg-L 1 -L 2 -L 3 -L 4 -D
    Formula VI
    其中，in,
    当L1的定义如权利要求1-4任一项所定义且不为时，Lg为和抗体反应时的离去基团；优选地，Lg选自卤素、砜基、三级胺盐基(Me3N+、Et3N+)、重氮盐基、-OMs、MeSO2-、CF3SO3-、对甲苯磺酰基；更优选地，Lg选自F、Cl、Br、MeSO2-；特别优选地，Lg为MeSO2-；L2、L3、L4和D的定义如权利要求1-4任一项所定义；When L 1 is defined as defined in any one of claims 1 to 4 and is not When , Lg is the leaving group when reacting with the antibody; preferably, Lg is selected from halogen, sulfone group, tertiary amine base (Me 3 N + , Et 3 N + ), diazonium base, -OMs, MeSO 2 -, CF 3 SO 3 -, p-toluenesulfonyl group; more preferably, Lg is selected from F, Cl, Br, MeSO 2 -; particularly preferably, Lg is MeSO 2 -; L 2 , L 3 , L 4 and D is defined as defined in any one of claims 1-4;
    当L1为时，Lg-L1为L2、L3、L4和D的定义如权利要求1-4任一项所定义。When L 1 is When , Lg-L 1 is L 2 , L 3 , L 4 and D are as defined in any one of claims 1 to 4.
  14. 如权利要求12或13所述的药物连接体偶联物或所述药物连接体偶联物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物，特征在于，所述的药物连接体偶联物的结构如式VII所示，
    The drug linker conjugate of claim 12 or 13 or the stereoisomer of the drug linker conjugate, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate It is characterized in that the structure of the drug linker conjugate is as shown in formula VII,
    其中，Lg、L1、L2、Rx1、Ry1和D的定义如权利要求13所定义。Wherein, Lg, L 1 , L 2 , R x1 , R y1 and D are defined as in claim 13 .
  15. 如权利要求12-14任一项所述的药物连接体偶联物、其立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物，特征在于所述药物连接体偶联物选自：







    The drug linker conjugate, its stereoisomer, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate according to any one of claims 12 to 14, characterized in that The drug linker conjugate is selected from:







  16. 一种制备根据权利要求1-11中任一项所述的式I所示的抗体药物偶联物的方法，其包括将Tb分别与根据权利要求12所述的式V所示的药物连接体偶联物在溶剂中进行偶联反应形成C-S键的步骤；A method for preparing the antibody-drug conjugate represented by formula I according to any one of claims 1-11, which includes combining Tb with the drug linker represented by formula V according to claim 12 The step in which the conjugate undergoes a coupling reaction in a solvent to form a C-S bond;
    所述的Tb与式V所示的药物连接体偶联物的摩尔比为1:(1-20)，如1:(2-20)、1:(4-20)、1:(6-20)、1:(8-20)、1:(10-20)、1:(12-20)、1:(14-20)、1:(16-20)或1:(18-20)；The molar ratio of the Tb to the drug linker conjugate represented by Formula V is 1:(1-20), such as 1:(2-20), 1:(4-20), 1:(6- 20), 1:(8-20), 1:(10-20), 1:(12-20), 1:(14-20), 1:(16-20) or 1:(18-20) ;
    所述的偶联反应优选为在水和/或有机溶剂中进行；所述的有机溶剂优选为N,N-二甲基甲酰胺、N,N-二甲基乙酰胺、二甲基亚砜、N-甲基吡咯烷酮中的一种或多种。The coupling reaction is preferably carried out in water and/or an organic solvent; the organic solvent is preferably N,N-dimethylformamide, N,N-dimethylacetamide, or dimethyl sulfoxide. , one or more of N-methylpyrrolidone.
  17. 药物组合物，其包含如权利要求1-11任一项所述的抗体药物偶联物、或所述抗体药物偶联物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物；或者如权利要求12-15任一项所述的药物连接体偶联物、其立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物以及任选的一种或多种药用辅料。A pharmaceutical composition comprising the antibody drug conjugate of any one of claims 1 to 11, or a stereoisomer of the antibody drug conjugate, a prodrug thereof, a pharmaceutically acceptable salt thereof, or Its pharmaceutically acceptable solvate; or the drug linker conjugate according to any one of claims 12-15, its stereoisomer, its prodrug, its pharmaceutically acceptable salt or its pharmaceutical acceptable solvates and optionally one or more pharmaceutical excipients.
  18. 如权利要求1-11任一项所述的抗体药物偶联物、或所述抗体药物偶联物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物；或者如权利要求12-15任一项所述的药物连接体偶联物、或所述药物连接体偶联物的立体异构体、其前药、其药学上可接受的盐或其药学上可接受的溶剂合物；或者前述权利要求17的药物组合物在制备治疗和/或预防实体肿瘤或血液肿瘤的药物中的用途，优选地所述肿瘤选自食管癌、脑瘤、肺癌、鳞状上皮细胞癌、膀胱癌、胃癌、卵巢癌、腹膜癌、胰腺癌、乳腺癌、头颈癌、子宫颈癌、子宫内膜癌、结 直肠癌、肝癌、肾癌、尿路上皮癌、实体瘤、非霍奇金淋巴瘤、中枢神经系统肿瘤、前列腺癌和甲状腺癌中的一种或多种。 The antibody drug conjugate according to any one of claims 1 to 11, or the stereoisomer of the antibody drug conjugate, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable salt Solvate; or the drug linker conjugate according to any one of claims 12 to 15, or the stereoisomer of the drug linker conjugate, its prodrug, or its pharmaceutically acceptable salt Or its pharmaceutically acceptable solvate; or the use of the pharmaceutical composition of claim 17 in the preparation of drugs for the treatment and/or prevention of solid tumors or hematological tumors, preferably the tumors are selected from esophageal cancer, brain tumors , lung cancer, squamous cell carcinoma, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, tuberculosis One or more of rectal cancer, liver cancer, renal cancer, urothelial cancer, solid tumors, non-Hodgkin lymphoma, central nervous system tumors, prostate cancer, and thyroid cancer.
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CN101287498A (en) * 2005-06-20 2008-10-15 Psma开发有限公司 PSMA antibody-drug conjugates
CN102264720A (en) * 2008-10-29 2011-11-30 细胞基因公司 Isoindoline compounds for use in the treatment of cancer
CN103396397A (en) * 2013-08-14 2013-11-20 中国人民解放军军事医学科学院毒物药物研究所 Lenalidomide derivative and use thereof as medicine
US20160082120A1 (en) * 2014-09-23 2016-03-24 Genentech, Inc. METHODS OF USING ANTI-CD79b IMMUNOCONJUGATES
CN106456794A (en) * 2014-05-22 2017-02-22 斯索恩生物制药有限公司 Site-specific conjugation of linker drugs to antibodies and resulting adcs
WO2021004391A1 (en) * 2019-07-08 2021-01-14 苏州开拓药业股份有限公司 C-myc protein inhibitor, and preparation method therefor and use thereof
WO2021069705A1 (en) * 2019-10-09 2021-04-15 Monte Rosa Therapeutics Isoindolinone compounds

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101287498A (en) * 2005-06-20 2008-10-15 Psma开发有限公司 PSMA antibody-drug conjugates
CN102264720A (en) * 2008-10-29 2011-11-30 细胞基因公司 Isoindoline compounds for use in the treatment of cancer
CN103396397A (en) * 2013-08-14 2013-11-20 中国人民解放军军事医学科学院毒物药物研究所 Lenalidomide derivative and use thereof as medicine
CN106456794A (en) * 2014-05-22 2017-02-22 斯索恩生物制药有限公司 Site-specific conjugation of linker drugs to antibodies and resulting adcs
US20160082120A1 (en) * 2014-09-23 2016-03-24 Genentech, Inc. METHODS OF USING ANTI-CD79b IMMUNOCONJUGATES
WO2021004391A1 (en) * 2019-07-08 2021-01-14 苏州开拓药业股份有限公司 C-myc protein inhibitor, and preparation method therefor and use thereof
WO2021069705A1 (en) * 2019-10-09 2021-04-15 Monte Rosa Therapeutics Isoindolinone compounds

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