WO2022022649A1 - 一种双药链接组装单元及双药靶向接头-药物偶联物 - Google Patents

一种双药链接组装单元及双药靶向接头-药物偶联物 Download PDF

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WO2022022649A1
WO2022022649A1 PCT/CN2021/109337 CN2021109337W WO2022022649A1 WO 2022022649 A1 WO2022022649 A1 WO 2022022649A1 CN 2021109337 W CN2021109337 W CN 2021109337W WO 2022022649 A1 WO2022022649 A1 WO 2022022649A1
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drug
dual
linker
drug conjugate
targeting linker
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PCT/CN2021/109337
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English (en)
French (fr)
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黄金昆
吴成龙
冯超阳
鲁岳
耿嘉豪
李磊
徐鑫
刘俊强
张伟
陈方
谢德建
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成都科岭源医药技术有限公司
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Publication of WO2022022649A1 publication Critical patent/WO2022022649A1/zh

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    • 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
    • A61K47/54—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 organic compound
    • A61K47/55—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 organic compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • 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
    • A61K47/54—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 organic compound
    • 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
    • A61K47/54—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 organic compound
    • A61K47/545—Heterocyclic compounds
    • A—HUMAN NECESSITIES
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    • 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
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    • A61K47/549—Sugars, nucleosides, nucleotides or nucleic acids
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    • 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
    • A61K47/62—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 a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • 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
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    • 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
    • A61K47/62—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 a protein, peptide or polyamino acid
    • A61K47/65—Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers
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    • 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
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    • 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
    • A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
    • A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • 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
    • A61K47/6835—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 the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
    • A61K47/6851—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 the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a determinant of a tumour cell
    • A—HUMAN NECESSITIES
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Definitions

  • the drugs contained in ADCs recognized by the FDA to date mainly target DNA or tubulin.
  • the marketed ADCs are respectively linked by antibodies and drugs targeting DNA or tubulin.
  • U is substituted or unsubstituted aryl, heteroaryl, alkane, cycloalkyl or heterocycloalkyl; the substituent is selected from deuterium, halogen, cyano, hydroxyl, C 1-6 alkyl or C 1-6 alkoxy;
  • X 1 and X 2 are independently selected from Wherein, a, b, c and d are each independently selected from 0 or 1, K 3 is selected from unsubstituted, unsubstituted or C 1-4 alkylene substituted by R 1 , and K 4 is selected from unsubstituted, unsubstituted or substituted by R 1 .
  • B 1 , B 2 , C 1 , C 2 , E 1 , E 2 are each independently selected from substituted or unsubstituted following groups: L 2a CONHL 3 , L 4 OL 5 CO or none; the substituents are each independently selected from C 1-5 alkyl; wherein L 2a , L 3 , L 4 , L 5 , L 6 are each independently selected from 0-2 methylene groups;
  • X 1 and X 2 are independently selected from Wherein, a, b, c, d are each independently selected from 0 or 1, R 1 , R 2 , R 3 , R 4 are each independently selected from H, C 1-5 alkyl, substituted or unsubstituted benzyl , -L 1a NHCONH 2 , L 1a is 0-3 methylene groups; the substituent on the benzyl group is selected from deuterium, halogen, cyano, hydroxyl, C 1-6 alkyl or C 1-6 alkoxy base;
  • the structure of the double drug linkage assembly unit is selected from one of the following structures:
  • the structure of the double drug link assembly unit is one of the following structures:
  • D 1 and D 2 are as described above.
  • the double drug link assembly unit is one of the following structures:
  • Ab is a targeting linker
  • q is an integer from 1 to 20
  • T, U, L1, L2, D1, and D2 are as described above.
  • the targeting linker is an antibody, antibody fragment, protein or nucleic acid aptamer, and the antibody is preferably an antibody against cell surface receptors and tumor-associated antigens;
  • the said q is an integer of 1-8.
  • the structure of the dual-drug targeting linker-drug conjugate molecule is selected from one of the following structures:
  • the present invention also provides a dual-drug targeting linker-drug conjugate, or a stereoisomer thereof, or an optical isomer thereof, wherein the dual-drug targeting linker-drug conjugate is composed of a targeting linker and The above-mentioned dual-drug linkage assembly units are connected;
  • the targeting linker is a substance that can target and bind to the diseased site, preferably an antibody, an antibody fragment, a protein or a nucleic acid aptamer; the antibody is preferably directed against cell surface receptors and Antibodies to tumor-associated antigens.
  • the dual-drug targeting linker-drug conjugate is composed of two or more of the dual-drug targeting linker-drug conjugate molecules shown in formula IIa, and the dual-drug targeting linker-drug conjugate is The DAR value of the conjugate is 6.00-7.00, preferably 6.41;
  • the dual-drug targeting linker-drug conjugate is composed of two or more of the dual-drug targeting linker-drug conjugate molecules shown in formula IIc, and the dual-drug targeting linker-drug conjugate
  • the DAR value of the material is 4.00 to 5.00, preferably 4.38;
  • the dual-drug targeting linker-drug conjugate is composed of two or more of the dual-drug targeting linker-drug conjugate molecules shown in formula IIe, and the dual-drug targeting linker-drug conjugate is The DAR value of the material is 3.00 to 4.00, preferably 3.35;
  • the dual-drug targeting linker-drug conjugate is composed of two or more of the dual-drug targeting linker-drug conjugate molecules shown in the formula IIf, and the dual-drug targeting linker-drug conjugate is The DAR value of the material is 2.00 to 3.00, preferably 2.31;
  • the dual-drug targeting linker-drug conjugate is composed of two or more of the dual-drug targeting linker-drug conjugate molecules shown in formula II SMP-52581, and the dual-drug targeting linker-
  • the DAR value of the drug conjugate is 1.50-2.50, preferably 1.93;
  • the dual-drug targeting linker-drug conjugate is composed of two or more of the dual-drug targeting linker-drug conjugate molecules shown in formula II SMP-96745, and the dual-drug targeting linker-
  • the DAR value of the drug conjugate is 4.00-5.00, preferably 4.27;
  • the dual-drug targeting linker-drug conjugate is composed of two or more of the dual-drug targeting linker-drug conjugate molecules shown in formula II SMP-93954, and the dual-drug targeting linker-
  • the DAR value of the drug conjugate is 4.00-5.00, preferably 4.32;
  • the dual-drug targeting linker-drug conjugate is composed of two or more of the dual-drug targeting linker-drug conjugate molecules shown in formula II SMP-11115, and the dual-drug targeting linker-
  • the DAR value of the drug conjugate is 7.00-8.00, preferably 7.71;
  • the dual-drug targeting linker-drug conjugate is composed of two or more of the dual-drug targeting linker-drug conjugate molecules shown in formula II SMP-04404, and the dual-drug targeting linker-
  • the DAR value of the drug conjugate is 4.00-5.00, preferably 4.67;
  • the dual-drug targeting linker-drug conjugate is composed of two or more of the dual-drug targeting linker-drug conjugate molecules shown in formula II SMP-37241, and the dual-drug targeting linker-
  • the DAR value of the drug conjugate is 6.00-7.00, preferably 6.39;
  • the dual-drug targeting linker-drug conjugate is composed of two or more of the dual-drug targeting linker-drug conjugate molecules shown in formula II SMP13432, and the dual-drug targeting linker-drug conjugate is The DAR value of the conjugate is 4.00-5.00, preferably 4.26;
  • the dual-drug targeting linker-drug conjugate is composed of two or more of the dual-drug targeting linker-drug conjugate molecules shown in formula II SMP-84984, and the dual-drug targeting linker-
  • the DAR value of the drug conjugate is 5.00-6.00, preferably 5.69;
  • the dual-drug targeting linker-drug conjugate is composed of two or more of the dual-drug targeting linker-drug conjugate molecules shown in formula II SMP-76794, and the dual-drug targeting linker-
  • the DAR value of the drug conjugate is 2.00-3.00, preferably 2.58;
  • the dual-drug targeting linker-drug conjugate is composed of two or more of the dual-drug targeting linker-drug conjugate molecules shown in formula II SMP-99767, and the dual-drug targeting linker-
  • the DAR value of the drug conjugate is 6.50-7.50, preferably 7.14;
  • the dual-drug targeting linker-drug conjugate is composed of two or more of the dual-drug targeting linker-drug conjugate molecules shown in formula II SMP-86145, and the dual-drug targeting linker-
  • the DAR value of the drug conjugate is 6.00-7.00, preferably 6.45;
  • the dual-drug targeting linker-drug conjugate is composed of two or more of the dual-drug targeting linker-drug conjugate molecules shown in formula II SMP-54418, and the dual-drug targeting linker-
  • the DAR value of the drug conjugate is 7.00-8.00, preferably 7.26;
  • the dual-drug targeting linker-drug conjugate is composed of two or more of the dual-drug targeting linker-drug conjugate molecules shown in formula II SMP-44708, and the dual-drug targeting linker-
  • the DAR value of the drug conjugate is 1.50-2.50, preferably 2.07.
  • the present invention also provides the above-mentioned dual-drug targeting linker-drug conjugate molecule, or a method for preparing the above-mentioned dual-drug targeting linker-drug conjugate, the method comprising the following steps: linking and assembling the targeting linker and the dual-drug conjugate
  • the unit is coupled, purified, and obtained; preferably, the purification method is dialysis.
  • the present invention also provides a medicine for preventing and/or treating tumors, which is based on the above-mentioned dual-drug targeting linker-drug conjugate molecule, or its stereoisomer, or its optical isomer, or the above-mentioned dual-drug targeting linker-drug conjugate molecule.
  • the drug-targeting linker-drug conjugate, or its stereoisomer or its optical isomer is the active ingredient, and the preparation is prepared by adding pharmaceutically acceptable excipients.
  • the present invention also provides the aforementioned dual-drug targeting linker-drug conjugate molecule, or its stereoisomer, or its optical isomer, or the aforementioned dual-drug targeting linker-drug conjugate, or its stereoisomer Use of the isomer or its optical isomer in the preparation of a medicament for preventing and/or treating tumors.
  • the tumor is selected from lung cancer, urethral cancer, colorectal cancer, prostate adenocarcinoma, ovarian cancer, pancreatic cancer, breast cancer, bladder cancer, gastric cancer, gastrointestinal stromal tumor, cervical cancer, esophageal cancer, squamous cell cancer, peritoneal cancer, liver cancer, colon cancer, rectal cancer, colorectal cancer, uterine cancer, salivary gland cancer, kidney cancer, vulvar cancer, thyroid cancer, penile cancer, leukemia, malignant lymphoma, plasmacytoma, myeloma or sarcoma.
  • Antibody or “antibody unit” is within its scope and includes any portion to which an antibody binds. This unit may bind, reactively associate or complex a receptor, antigen, or other receptor unit possessed by the targeted cell population.
  • An antibody can be any protein or intraprotein molecule that can bind, complex or react with a portion of the cell population to be treated or bioengineered.
  • the antibody in the present invention can specifically bind to the antigen.
  • the designed antigens include: tumor-associated antigens (TAAs), cell surface receptor proteins and other cell surface molecules, cell survival regulators, cell proliferation regulators, molecules associated with tissue growth or differentiation (such as known or predicted functional sex), lymphokines, cytokines, factors involved in the regulation of the cell cycle, molecules involved in angiogenesis, and molecules related to angiogenesis.
  • TAAs tumor-associated antigens
  • cell survival regulators such as known or predicted functional sex
  • lymphokines cytokines
  • factors involved in the regulation of the cell cycle molecules involved in angiogenesis
  • molecules related to angiogenesis e.g, CD protein.
  • the antigens to which the antibodies of the present invention bind can be one or a subset of the above categories, other subsets include other molecules/antigens with specific properties.
  • Antibodies used in the dual-drug linkage assembly unit and dual-drug targeting linker-drug conjugates of the present invention include, but are not limited to, antibodies against tumor-associated antigens of cell surface receptors.
  • Tumor-associated antigens include, but are not limited to, the tumor-associated antigens listed below, including names and GenBank accession numbers.
  • Antibodies targeting corresponding tumor-associated antigens include all amino acid sequence variants and isotypes that are at least 70%, 80%, 85%, 90%, or 95% homologous to sequences identified in references, or have The tumor-associated antigen sequences in the literature have completely identical biological properties and properties.
  • Tumor-associated antigens are: BMPR1B (Genbank accession number: NM-001203), E16 (Genbank accession number: NM-003486), STEAP1 (Genbank accession number: NM-012449), 0772P (Genbank accession number: AF361486), MPF (Genbank accession number: AF361486) Accession number: NM-005823), Napi3b (Genbank accession number: NM-006424), Sema 5b (Genbank accession number: AB040878), PSCA hlg (Genbank accession number: AY358628), ETBR (Genbank accession number: AY275463), MSG783 ( Genbank accession number: NM-017763), STEAP2 (Genbank accession number: AF455138), TrpM4 (Genbank accession number: NM-017636), CRIPTO (Genbank accession number: NP-003203 or NM-003212), CD21 (Genbank accession number
  • drug generally refers to any compound having the desired biological activity and having reactive functional groups to prepare the conjugates of the present invention.
  • the drug includes a cytotoxic compound for cancer treatment, a biologically active protein or polypeptide, including but not limited to camptothecin derivatives, such as SN-38, Dxd, Dx-8951, which act on microbes.
  • camptothecin derivatives such as SN-38, Dxd, Dx-8951, which act on microbes.
  • Compounds of tubulin such as Eribulin, MMAE, MMAF, maytansine, etc. (structure below).
  • linkers or “antibody drug conjugate linkers” described herein can be divided into two categories: non-cleavable linkers and cleavable linkers.
  • the drug release mechanism is: after the conjugate is bound to the antigen and is endocytosed by cells, the antibody is enzymatically hydrolyzed in the lysosome to release the small molecule drug, Active small molecules composed of linkers and antibody amino acid residues.
  • Cleavable linkers break within the target cell and release the active drug (the small molecule drug itself), and cleavable linkers can be divided into two main categories: chemically labile linkers and enzymatically labile linkers.
  • Chemically labile linkers can be selectively cleaved due to differences in plasma and cytoplasmic properties, including pH, glutathione concentration, etc. Enzyme-labile linkers, such as peptide linkers, can better control drug release. Peptide linkers can be efficiently cleaved by lysosomal proteases such as cathepsin or plasmin. This peptide linkage is believed to be very stable in plasma because proteases are generally inactive outside the cell due to unfavorable extracellular pH and serum protease inhibitors. Given the high plasma stability and good intracellular cleavage selectivity and effectiveness, enzyme-labile linkers are widely used as cleavable linkers for antibody-drug conjugates.
  • q represents the number of dual-drug linking assembly units coupled on one targeting linker in the dual-drug targeting linker-drug conjugate molecule, and q is an integer.
  • the dual-drug targeting linker-drug conjugate used in the art is usually a mixture composed of multiple dual-drug targeting linker-drug conjugate molecules with different q values.
  • DAR value represents the average number of dual-drug linking assembly units coupled to one targeting linker in the dual-drug targeting linker-drug conjugate, which is equivalent to the average of the q values. DAR values may not be integers.
  • the present invention provides a dual-drug linking assembly unit, which can be connected with a targeting linker to obtain a corresponding dual-drug targeting linker-drug conjugate, and the dual-drug targeting linker-drug conjugate can Targeting tumor cells, reducing the toxic and side effects on normal cells, and effectively overcoming drug resistance.
  • the ADC provided by the present invention has significantly improved inhibitory effects on breast adenocarcinoma cells and gastric cancer cells.
  • the dual-drug linking assembly unit and the dual-drug targeting linker-drug conjugate of the present invention have broad application prospects in the preparation of preventive and/or therapeutic anti-tumor drugs.
  • Examples 1 to 22 are specific preparation examples of the dual drug link assembly unit of the present invention.
  • Step 3 Preparation of target product XIe:
  • Step 1 Preparation of target product XIc:
  • Step 2 Preparation of the target product SMP-52581:
  • Step 1 Preparation of the target product SPM-96745:
  • Step 2 Preparation of target product XIf:
  • Step 1 Preparation of the target product SMP-39545:
  • Step 2 Preparation of target product SMP-93954:
  • Step 1 Preparation of the target product SMP-11115:
  • Step 1 Preparation of the target product SMP-04404:
  • Step 3 Preparation of the target product SMP-37241:
  • Step 1 Preparation of the target product SMP-13432:
  • Step 1 Preparation of the target product SMP-84984:
  • Step 2 Preparation of target product XIa:
  • Step 3 Preparation of the target product SMP-04140:
  • Step 1 Preparation of the target product SMP-76794:
  • Step 1 Preparation of the target product SMP-99767:
  • Step 2 Preparation of the target product SMP-86145:
  • Step 5 Preparation of the target product SMP-54418:
  • Step 2 Preparation of the target product SMP-44708:
  • Example 23 is a specific preparation example of the antibody-drug conjugate (ADC) of the present invention.
  • the corresponding ADCs were prepared by using the dual-drug linkage assembly units and antibodies prepared in Examples 1 to 22 as raw materials, respectively. Its general method is as follows:
  • Trastuzumab was prepared as a 10 mg/mL solution (3.0 mL) in PBS6.0/EDTA, to which was added 10 mM tris(2-carboxyethyl)phosphine hydrochloride (TCEP) aqueous solution (0.0934 mL) and 1 M aqueous dipotassium hydrogen phosphate solution (0.150 mL), ensuring that the pH of the reaction solution is between 7.3-7.5, stirred for 1 minute and incubated at 37°C for 1 hour.
  • TCEP tris(2-carboxyethyl)phosphine hydrochloride
  • Dialyze the reaction solution obtained in step 2 with a 10KD semipermeable membrane the dialysis temperature is 25°C, the dialysis solvent is 4L/time, the dialysis time is 4 hours/time, and the dialysate is dialyzed 4 times in total.
  • the obtained dialysate is centrifuged and concentrated to ADC using a 10KD ultrafiltration centrifuge tube. The concentration is about 5mg/mL, and it is stored at -20°C ⁇ -30°C for future use.
  • Example 1-ADC The prepared ADCs were named as Example 1-ADC to Example 22-ADC, respectively, and the structures were as follows:
  • Each ADC sample prepared in Example 23 was diluted to 1 mg/ml with pH 8.0, 50 mmol/L Tris buffer, and then freshly prepared DTT stock solution (1 mol/L) was added to make dithiothreitol ( DTT) with a final concentration of 50 mmol/L, incubate at 37°C for 20-30 min, and carry out the reduction reaction.
  • Step 2 DAR value determination
  • step 1 The sample obtained in step 1 was injected into LC-MS for detection, and after deconvolution processing, the HPLC peak areas of L+0, L+1, H+0, H+1, H+2, H+3, H+4 were obtained, The DAR value is obtained by calculation.
  • the DAR value here represents the average number of dual-drug linkage assembly units coupled to an antibody in the ADC sample to be tested.
  • Example 17 - ADC 7.14 Example 13 - ADC 5.69 Example 1 - ADC 3.35 Example 2 - ADC 4.38 Example 4 - ADC 4.27 Example 12 - ADC 4.26 Example 15 - ADC 2.47 Example 16 - ADC 2.58 Example 18 - ADC 6.45 Example 19 - ADC 7.26 Example 20 - ADC 2.07 Example 21 - ADC 1.99 Example 22 - ADC 3.47
  • SK-BR-3 cells human breast adenocarcinoma cells
  • N87 cells human gastric cancer cells
  • each ADC to be tested ie Example 1-ADC to Example 22-ADC
  • the positive control Trastuzumab-GGFG-Dxd DS-8201a, a known ADC
  • DS-8201a a known ADC
  • the positive control Cisplatin (cisplatin)
  • the prepared ADCs to be tested and the positive control were added to the cells, and the medium-only well was used as the negative control.
  • the cells were cultured for 144 hours after adding the drug, and then the cell viability was detected by CTG (CELL TITER-GLO) method, and the IC50 values of each tested drug on SK-BR-3 cells and N87 cells were calculated respectively. The results are shown in Table 2 below.
  • the ADC provided by the present invention can significantly improve the inhibitory effect on breast adenocarcinoma cells and gastric cancer cells.
  • the inhibitory effect of the ADC provided by the present invention on breast adenocarcinoma cells and gastric cancer cells is also significantly improved.
  • the present invention provides a dual-drug linking assembly unit, which can be connected with a targeting linker to obtain a corresponding dual-drug targeting linker-drug conjugate, the dual-drug targeting linker-drug conjugate.
  • the conjugate can target tumor cells, reduce the toxic and side effects on normal cells, and can effectively overcome drug resistance.
  • the ADC provided by the present invention has significantly improved inhibitory effects on breast adenocarcinoma cells and gastric cancer cells.
  • the dual-drug linking assembly unit and the dual-drug targeting linker-drug conjugate of the present invention have broad application prospects in the preparation of preventive and/or therapeutic anti-tumor drugs.

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Abstract

一种双药链接组装单元及双药靶向接头-药物偶联物。具体提供了一种式（VI）所示的双药物链接组装单元、或其立体异构体、或其旋光异构体。该双药链接组装单元能够与靶向接头连接得到式（I）所示的双药靶向接头-药物偶联物分子。该双药靶向接头-药物偶联物能够靶向作用于肿瘤细胞，降低对正常细胞的毒副作用，同时能够有效克服药物的耐药性。与已经上市的ADC：DS-8201a相比，提供的ADC对乳腺腺癌细胞和胃癌细胞的抑制作用都明显提高，以上双药链接组装单元和双药靶向接头-药物偶联物在制备预防和/或治疗的抗肿瘤药物中具有广阔的应用前景。

Description

一种双药链接组装单元及双药靶向接头-药物偶联物 技术领域
本发明属于药物领域，具体涉及一种双药物链接组装单元及其对应的双药靶向接头-药物偶联物。
背景技术
抗体-药物偶联物(Antibody-Drug Conjugate；ADC)能够选择性的向癌细胞输送药物，并杀死癌细胞，但是对正常细胞的影响较小，开创了肿瘤治疗的新纪元。作为ADC，已有多个药物获得FDA批准上市，如抗体CD33连接卡奇霉素而成的Mylotarg，CD30抗体连接澳瑞他汀E而成的Adcetris用于霍奇金淋巴瘤和未分化大细胞淋巴瘤患者的治疗，Her2抗体和喜树碱衍生物Dxd连接而成的DS-8201用于HER2阳性的乳腺癌患者的治疗以及靶向TROP-2抗原(也称作上皮糖蛋白1，EGP-1)的Sacituzumab govitecan。
迄今为止被FDA承认的ADC中含有的药物主要以DNA或微管蛋白为靶标。已上市的ADC分别由抗体和靶向DNA或者微管蛋白的药物连接，目前没有任何将抗体和靶向DNA和微管蛋白的药物同时连接的ADC上市，尚不清楚将靶向DNA和微管蛋白的药物同时连接到抗体上是否能通过两种不同的抗肿瘤作用机制杀伤肿瘤，是否能有效发挥作用。
作为抗肿瘤的小分子化合物，已知作为抑制DNA拓扑异构酶I而达到抗肿瘤作用的化合物的喜树碱衍生物，如SN-38,Dxd，Dx-8951在体内和体外均确认了对多种癌细胞有杀伤作用，显示出强抗肿瘤效果。作为抑制微管蛋白而达到抗肿瘤作用的化合物，如Eribulin，MMAE，MMAF，美登素等在体内和体外均确认了对多种癌细胞有杀伤作用，显示出强抗肿瘤效果。将两种不同机制的抗肿瘤药物连接到相同的抗体上，可能会对抗肿瘤活性有协同作用，也可能会抗肿瘤活性产生拮抗作用，实际效果无法预期。
因此，亟需研究出一种高效、安全的多药物多靶点作用机制的ADC，这对开发抗肿瘤效果和安全性优异的抗肿瘤药物具有非常重要的意义。
发明内容
本发明的目的在于提供一种双药物链接组装单元及其对应的双药靶向接头-药物偶联物。
本发明提供了式VI所示的双药物链接组装单元、或其立体异构体、或其旋光异构体：
Figure PCTCN2021109337-appb-000001
其中，T是拴系基团，它能够与靶向接头连接；所述靶向接头为能够靶 向结合病变部位的物质；
U是三叉型连接子部分；
L1和L2为可断裂或不可断裂的链接基团；
D 1、D 2分别是第一药物单元和第二药物单元。
进一步地，所述T能够与靶向接头上的巯基反应连接。
进一步地，所述双药物链接组装单元的结构如式VI-1所示：
Figure PCTCN2021109337-appb-000002
其中，K 1、K 2各自独立的选自无、K aK bK c；K a为苯基，K b选自O、S或无，K c选自0～2个亚甲基；
n，p各自独立的选自0～10的整数；
U为取代或未取代的芳基、杂芳基、链烷基、环烷基或杂环烷基；所述取代基选自氘、卤素、氰基、羟基、C 1～6烷基或C 1～6烷氧基；
X 1、X 2各自独立的选自
Figure PCTCN2021109337-appb-000003
其中，a、b、c、d各自独立的选自0或1，K 3选自无、无取代或被R 1取代的C 1～4亚烷基，K 4选自无、无取代或被R 2取代的C 1～4亚烷基，K 5选自无、无取代或被R 3取代的C 1～4亚烷基，K 6选自无、无取代或被R 4取代的C 1～4亚烷基，R 1、R 2、R 3、R 4各自独立的选自H、C 1～5烷基、取代或者未取代的苄基、-L 1aNHCONH 2、-L 1aNHCOCR 5NH 2，L 1a为0～4个亚甲基，R 5为C 1～3烷基；所述苄基上的取代基选自氘、卤素、氰基、羟基、C 1～6烷基或C 1～6烷氧基；
B 1，B 2，C 1，C 2，E 1，E 2各自独立的选自取代或未取代的以下基团：
Figure PCTCN2021109337-appb-000004
L 2aCONHL 3、L 4OL 5CO或无；所述取代基各自独立的选自
Figure PCTCN2021109337-appb-000005
C 1～5烷基；其中L 2a、L 3、L 4、L 5、L 6各自独立的选自0～2个亚甲基；
D 1、D 2各自独立的选自细胞毒性药物、治疗自身免疫疾病的药物或抗炎的药物；
T如上所述。
进一步地，所述双药物链接组装单元的结构如式VII所示：
Figure PCTCN2021109337-appb-000006
其中，n，p各自独立的选自0～10的整数；
U为取代或未取代的芳基、杂芳基、链烷基、环烷基或杂环烷基；所述取代基选自氘、卤素、氰基、羟基、C 1～6烷基或C 1～6烷氧基；
X 1、X 2各自独立的选自
Figure PCTCN2021109337-appb-000007
其中，a、b、c、d各自独立的选自0或1，R 1、R 2、R 3、R 4各自独立的选自H、C 1～5烷基、取代或者未取代的苄基、-L 1aNHCONH 2，L 1a为0～3个亚甲基；所述苄基上的取代基选自氘、卤素、氰基、羟基、C 1～6烷基或C 1～6烷氧基；
B 1，B 2，C 1，C 2，E 1，E 2各自独立的选自取代或未取代的以下基团：
Figure PCTCN2021109337-appb-000008
L 2aCONHL 3、L 4OL 5CO或无；所述取代基各自独立的选自
Figure PCTCN2021109337-appb-000009
C 1～5烷基；其中L 2a、L 3、L 4、L 5、L 6各自独立的选自0～2个亚甲基；
D 1、D 2各自独立的选自细胞毒性药物、治疗自身免疫疾病的药物或抗炎的药物；
T如上所述。
进一步地，所述双药物链接组装单元的结构如式VII-1所示：
Figure PCTCN2021109337-appb-000010
其中，A 1、A 2各自独立的选自0～10个亚甲基，O，S，CO，NH，CONH，NHCO或无；
m，n，p，U，X 1，X 2，B 1，B 2，C 1，C 2，E 1，E 2，D 1、D 2如上所述。
进一步地，所述双药物链接组装单元的结构如式VIIIa或VIIIb所示：
Figure PCTCN2021109337-appb-000011
其中，A 1、A 2、m、n、p、X 1、X 2、B 1、B 2、C 1、C 2、E 1、E 2、D 1、D 2如上所述；
M为氢、氘、卤素、氰基、羟基、C 1～6烷基或C 1～6烷氧基。
进一步地，所述双药物链接组装单元的结构选自以下结构之一：
Figure PCTCN2021109337-appb-000012
Figure PCTCN2021109337-appb-000013
Figure PCTCN2021109337-appb-000014
其中，D 1、D 2如上所述，n、p为0～4的整数；优选地，n＝p。进一步地，所述双药物链接组装单元的结构为以下结构之一：
Figure PCTCN2021109337-appb-000015
Figure PCTCN2021109337-appb-000016
其中，D 1、D 2如上所述。
进一步地，所述D 1、D 2各自独立的选自以DNA为靶标的药物单元或以微管蛋白为靶标的药物单元；所述以DNA为靶标的药物单元优选为SN-38，Dxd，Dx-8951或其衍生物，和/或，所述以微管蛋白为靶标的药物单元优选为Eribulin，MMAE，MMAF，美登素或其衍生物。
进一步地，所述双药物链接组装单元为以下结构之一：
Figure PCTCN2021109337-appb-000017
Figure PCTCN2021109337-appb-000018
Figure PCTCN2021109337-appb-000019
Figure PCTCN2021109337-appb-000020
Figure PCTCN2021109337-appb-000021
Figure PCTCN2021109337-appb-000022
Figure PCTCN2021109337-appb-000023
Figure PCTCN2021109337-appb-000024
Figure PCTCN2021109337-appb-000025
本发明还提供了一种双药靶向接头-药物偶联物分子、或其立体异构体、或其旋光异构体，所述双药靶向接头-药物偶联物分子是由靶向接头和q个上述的双药物链接组装单元连接所得；所述靶向接头为能够靶向结合病变部位的物质；所述双药靶向接头-药物偶联物分子的结构如式I所示：
Figure PCTCN2021109337-appb-000026
其中，Ab为靶向接头，q为1～20的整数，T、U、L1、L2、D 1、D 2如上所述。
进一步地，所述靶向接头为抗体、抗体片段、蛋白或核酸适配体，所述抗体优选为针对细胞表面受体和肿瘤相关抗原的抗体；
和/或，所述q为1～8的整数。
进一步地，所述双药靶向接头-药物偶联物分子的结构选自以下结构之一：
Figure PCTCN2021109337-appb-000027
Figure PCTCN2021109337-appb-000028
Figure PCTCN2021109337-appb-000029
Figure PCTCN2021109337-appb-000030
Figure PCTCN2021109337-appb-000031
Figure PCTCN2021109337-appb-000032
Figure PCTCN2021109337-appb-000033
Figure PCTCN2021109337-appb-000034
Figure PCTCN2021109337-appb-000035
本发明还提供了一种双药靶向接头-药物偶联物、或其立体异构体、或其旋光异构体，所述双药靶向接头-药物偶联物是由靶向接头和上述的双药物链接组装单元连接所得；所述靶向接头为能够靶向结合病变部位的物质，优选为抗体、抗体片段、蛋白或核酸适配体；所述抗体优选为针对细胞表面受体和肿瘤相关抗原的抗体。
进一步地，所述双药靶向接头-药物偶联物由式IIa所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为6.00～7.00，优选为6.41；
或，所述双药靶向接头-药物偶联物由式IIb所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为1.50～2.50，优选为2.03；
或，所述双药靶向接头-药物偶联物由式IIc所示双药靶向接头-药物偶联 物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为4.00～5.00，优选为4.38；
或，所述双药靶向接头-药物偶联物由式IId所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为1.50～2.50，优选为1.99；
或，所述双药靶向接头-药物偶联物由式IIe所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为3.00～4.00，优选为3.35；
或，所述双药靶向接头-药物偶联物由式IIf所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为2.00～3.00，优选为2.31；
或，所述双药靶向接头-药物偶联物由式IIg所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为3.00～4.00，优选为3.47；
或，所述双药靶向接头-药物偶联物由式II SMP-52581所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为1.50～2.50，优选为1.93；
或，所述双药靶向接头-药物偶联物由式II SMP-96745所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为4.00～5.00，优选为4.27；
或，所述双药靶向接头-药物偶联物由式II SMP-39545所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为5.00～6.00，优选为5.24；
或，所述双药靶向接头-药物偶联物由式II SMP-93954所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为4.00～5.00，优选为4.32；
或，所述双药靶向接头-药物偶联物由式II SMP-11115所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为7.00～8.00，优选为7.71；
或，所述双药靶向接头-药物偶联物由式II SMP-04404所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为4.00～5.00，优选为4.67；
或，所述双药靶向接头-药物偶联物由式II SMP-37241所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为6.00～7.00，优选为6.39；
或，所述双药靶向接头-药物偶联物由式II SMP13432所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为4.00～5.00，优选为4.26；
或，所述双药靶向接头-药物偶联物由式II SMP-84984所示双药靶向接 头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为5.00～6.00，优选为5.69；
或，所述双药靶向接头-药物偶联物由式II SMP-04140所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为2.00～3.00，优选为2.47；
或，所述双药靶向接头-药物偶联物由式II SMP-76794所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为2.00～3.00，优选为2.58；
或，所述双药靶向接头-药物偶联物由式II SMP-99767所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为6.50～7.50，优选为7.14；
或，所述双药靶向接头-药物偶联物由式II SMP-86145所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为6.00～7.00，优选为6.45；
或，所述双药靶向接头-药物偶联物由式II SMP-54418所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为7.00～8.00，优选为7.26；
或，所述双药靶向接头-药物偶联物由式II SMP-44708所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为1.50～2.50，优选为2.07。
本发明还提供了上述双药靶向接头-药物偶联物分子，或上述双药靶向接头-药物偶联物的制备方法，所述方法包括以下步骤：将靶向接头与双药物链接组装单元偶联，纯化，即得；优选地，所述纯化方式为透析。
本发明还提供了一种预防和/或治疗肿瘤的药物，它是以上述的双药靶向接头-药物偶联物分子、或其立体异构体、或其旋光异构体，或上述双药靶向接头-药物偶联物、或其立体异构体、或其旋光异构体为活性成分，加上药学上可接受的辅料制成的制剂。
本发明还提供了上述的双药靶向接头-药物偶联物分子、或其立体异构体、或其旋光异构体，或上述双药靶向接头-药物偶联物、或其立体异构体、或其旋光异构体在制备预防和/或治疗肿瘤的药物中的用途。
进一步地，所述肿瘤选自肺癌、尿道癌、大肠癌、前列腺腺癌、卵巢癌、胰腺癌、乳腺癌、膀胱癌、胃癌、胃肠道间质瘤、宫颈癌、食道癌、鳞状细胞癌、腹膜癌、肝癌、结肠癌、直肠癌、结直肠癌、子宫癌、唾液腺癌、肾癌、外阴癌、甲状腺癌、阴茎癌、白血病、恶性淋巴瘤、浆细胞瘤、骨髓瘤或肉瘤。
关于本发明的使用术语的定义：除非另有说明，本文中基团或者术语提供的初始定义适用于整篇说明书的该基团或者术语；对于本文没有具体定义的术语，应该根据公开内容和上下文，给出本领域技术人员能够给予它们的含义。
“抗体”或“抗体单元”在其所属范围内，包括抗体结合的任何部分。这一单元可以结合，反应性关联或者络合一个受体，抗原，或者靶向细胞群体具有的其他受体单元。抗体可以是任何蛋白或蛋白内分子，它可以结合，络合或者与待治疗或生物改造的细胞群体的一部分反应。
本发明中的抗体能够专一性的与抗原结合。设计的抗原包括：肿瘤相关抗原(TAA)，细胞表面受体蛋白和其他细胞表面分子，细胞存活调节因子，细胞增殖调节因子，与组织生长或分化相关的分子(如已知或预知的具有功能性的)，淋巴因子，细胞因子，参与细胞循环调节的因子，参与血管生成的分子，以及与血管生成相关的分子。肿瘤相关的因子可以是簇分化因子(如CD蛋白)。本发明中所述抗体结合的抗原可以是上述分类中的一个或一个子集，其他的子集包含其他的具有特殊性质的分子/抗原。
应用在本发明双药物链接组装单元及双药靶向接头-药物偶联物中的抗体包括，但不局限于，针对细胞表面受体的肿瘤相关的抗原的抗体。肿瘤相关抗原包括但不局限于，以下列出的肿瘤相关抗原，包括名称和基因库登录号。抗体靶向对应肿瘤相关的抗原包括所有的氨基酸序列变种和同种，与参考文献中确认的序列具有至少70％，80％，85％，90％或95％的同源性，或者具备与引用文献中的肿瘤相关抗原序列具有完全一致的生物性质和特性。
肿瘤相关抗原有：BMPR1B(Genbank登录号：NM-001203)，E16(Genbank登录号：NM-003486),STEAP1(Genbank登录号：NM-012449),0772P(Genbank登录号：AF361486),MPF(Genbank登录号：NM-005823),Napi3b(Genbank登录号：NM-006424),Sema 5b(Genbank登录号：AB040878),PSCA hlg(Genbank登录号：AY358628),ETBR(Genbank登录号：AY275463),MSG783(Genbank登录号：NM-017763),STEAP2(Genbank登录号：AF455138),TrpM4(Genbank登录号：NM-017636),CRIPTO(Genbank登录号：NP-003203或NM-003212),CD21(Genbank登录号：M26004),CD79B(Genbank登录号：NM-000626),FcRH2(Genbank登录号：NM-030764),HER2(Genbank登录号：M11730),NCA(Genbank登录号：M18728),MDP(Genbank登录号：BC017023),IL20Rα(Genbank登录号：AF184971),Brevican(Genbank登录号：AF229053),EphB2R(Genbank登录号：NM-004442),GEDA(Genbank登录号：AY260763),BAFF-R(Genbank登录号：AF1164546),CD22(Genbank登录号：AK026467),CD79a(Genbank登录号：NP-001774.1),CXCR5(Genbank登录号：NP-001701.1),HLA-DOB(Genbank登录号：NP-002111.1),P2X5(Genbank登录号：NP-002552.2),CD72(Genbank登录号：NP-001773.1),LY64(Genbank登录号：NP-005573.1),FcRH1(Genbank登录号：NP-443170.1),IRTA2(Genbank登录号：NP-112571.1),TENB2(Genbank登录号：AF179274)。
如本文所用，“药物”、“药物单元”或者代号“D”泛指任何具有期望的生物活性、并具有反应性官能团以制备本发明所述的偶联物的化合物。进一步地，药物包括一种用于癌症治疗的细胞毒性化合物，一种具有生物活性的蛋 白或者多肽，包括但不限于喜树碱衍生物，如SN-38,Dxd，Dx-8951，作用于微管蛋白的化合物，如Eribulin，MMAE，MMAF，美登素等(结构如下)。
Figure PCTCN2021109337-appb-000036
按照细胞内药物释放机制，本文所述“连接子”或“抗体药物偶联物连接子”可被分为两类：不可断裂连接子和可断裂连接子。
对于含有不可断裂连接子的双药抗体药物偶联物，其药物释放机制为：偶联物和抗原结合并被细胞内吞后，抗体在溶酶体中被酶解，释放出小分子药物，连接子和抗体氨基酸残基共同组成的活性小分子。
可断裂连接子在目标细胞内断裂并释放出活性药物(小分子药物自身)，可断裂连接子可分为两个主要的类别：化学不稳定连接子和酶不稳定连接子。
化学不稳定连接子可以由于血浆和细胞质性质的不同而选择性的断裂，这样的性质包括PH值，谷胱甘肽浓度等。酶不稳定连接子，如肽连接子，能够更好的控制药物释放。肽连接子能够被溶酶体内蛋白酶，如组织蛋白酶活纤溶酶有效的切断。这种肽连接被认为在血浆中非常稳定，因为细胞外不合宜的PH值及血清蛋白酶抑制剂导致蛋白酶通常在细胞外不具备活性。鉴于较高的血浆稳定性和良好的细胞内断裂选择性和有效性，酶不稳定连接子被广泛用于抗体药物偶联物的可断裂连接子。
本发明中，“q”表示双药靶向接头-药物偶联物分子中，一个靶向接头上偶联的双药物链接组装单元个数，q为整数。
但是，本领域使用的双药靶向接头-药物偶联物通常是由多个q值不同的双药靶向接头-药物偶联物分子组成的混合物。
“DAR值”表示双药靶向接头-药物偶联物中，一个靶向接头上偶联的双药物链接组装单元的平均个数，相当于q值的平均值。DAR值可以不是整数。
本发明提供了一种双药物链接组装单元，该双药物链接组装单元能够与靶向接头连接得到对应的双药靶向接头-药物偶联物，该双药靶向接头-药物偶联物能够靶向作用于肿瘤细胞，降低对正常细胞的毒副作用，同时能够有效克服药物的耐药性。与已经上市的ADC：DS-8201a相比，本发明提供的ADC对乳腺腺癌细胞和胃癌细胞的抑制作用都明显提高。本发明的双药物链接组装单元和双药靶向接头-药物偶联物在制备预防和/或治疗的抗肿瘤药物中具有广阔的应用前景。
显然，根据本发明的上述内容，按照本领域的普通技术知识和惯用手段，在不脱离本发明上述基本技术思想前提下，还可以做出其它多种形式的修改、替换或变更。
以下通过实施例形式的具体实施方式，对本发明的上述内容再作进一步的详细说明。但不应将此理解为本发明上述主题的范围仅限于以下的实例。凡基于本发明上述内容所实现的技术均属于本发明的范围。
具体实施方式
本发明所用原料与设备均为已知产品，通过购买市售产品所得。
实施例1～实施例22是本发明双药物链接组装单元的具体制备实施例。
实施例1、化合物XIe的合成：
步骤1：中间体4-2的制备：
Figure PCTCN2021109337-appb-000037
15mL反应瓶中加入原料L-2(300mg),DCM(5mL)，在0℃下缓慢滴加草酰氯(0.3mL)室温搅拌反应30分钟。减压浓缩除去DCM，残余物加入DCM(20mL)；将5-氨基间苯二酸(350mg)溶于DMF(2mL)中后加入DIPEA(320mg)，在0℃下缓慢滴加上述的DCM溶液，室温搅拌反应2小时。HPLC纯化(H 2O：ACN＝10-90％)，得4-2纯品，重200mg。MS(EI)m/s：375.1(M+H)。
步骤2：中间体4-3的制备：
Figure PCTCN2021109337-appb-000038
50mL反应瓶中依次加入原料4-2(53mg),DMF(2.5mL)，HATU(54mg)，DIPEA(18mg)室温搅拌反应5分钟。将A原料(40mg)溶于DMF(1mL)后缓慢滴加到上述的反应液中，室温反应20分钟。HPLC纯化(H 2O：ACN＝10-90％)，得4-3纯品，重20mg。MS(EI)m/s：740.5(M/2+H)。
步骤3：目标产物XIe的制备：
Figure PCTCN2021109337-appb-000039
25mL反应瓶中依次加入原料4-3(15mg),DMF(1.5mL)，HATU(5.7mg)，DIPEA(2.5mg)室温搅拌反应1分钟后加入原料C(11.5mg)，室温反应20分钟。HPLC纯化(H 2O：ACN＝10-90％)，得XIe纯品，重17.7mg。MS(EI)m/s：1299.3(M/2+H)。 1H NMR(400MHz,DMSO-d6)δ9.87(m,3H),8.29–8.13(m,5H),7.78(m,2H),7.57–7.44(m,5H),7.42–7.24(m,10H),6.81–6.66(m,3H),6.16(m,2H),5.41–5.31(m,4H),5.19(qm,1H),5.17–5.09(m,5H),5.07–5.00(m,3H),4.91(m,1H),4.49–3.68(m,28H),3.68–3.50(m,3H),3.44–3.36(m,1H),3.35–3.19(m,11H),3.18–3.01(m,4H),2.91(d,J＝7.8Hz,6H),2.68(m,1H),2.62–2.25(m,13H),2.25–1.46(m,40H),1.45–1.22(m,4H),1.18(m,3H),1.09(d,J＝7.0Hz,3H),1.02(d,J＝8.0Hz,3H),0.98–0.90(m,15H),0.90–0.77(m,15H).
实施例2、化合物XIc的合成：
Figure PCTCN2021109337-appb-000040
步骤1：目标产物XIc的制备：
25mL反应瓶中依次加入原料4-3(15mg),DMF(1.5mL)，HATU(5.7mg)，DIPEA(2.5mg)室温搅拌反应1分钟后加入B原料(8.5mg)，室温反应20分钟。HPLC纯化(H2O：ACN＝10-90％)，得XIc纯品，重9.8mg。MS(EI)m/s：1016.5(M/2+H)。
实施例3、化合物SMP-52581的合成：
步骤1：中间体4-2-B的制备：
Figure PCTCN2021109337-appb-000041
50mL反应瓶中依次加入原料4-2(89mg),DMF(2.5mL)，HATU(90mg)，DIPEA(30mg)室温搅拌反应5分钟。将B原料(50mg)溶于DMF(1mL)后缓慢滴加到上述的反应液中，室温反应20分钟。HPLC纯化(H 2O：ACN＝10-90％)，得纯品4-2-B，重35mg。MS(EI)m/s：1197.2(M+H)。
步骤2：目标产物SMP-52581的制备：
Figure PCTCN2021109337-appb-000042
15mL反应瓶中依次加入原料4-2-B(15mg),DMF(1.5mL)，HATU(7.1mg)，DIPEA(3.2mg)室温搅拌反应5分钟。将C原料(14mg)溶于DMF(0.5mL)后缓慢滴加到上述的反应液中，室温反应20分钟。HPLC纯化(H 2O：ACN＝10-90％)，得纯品，重6.9mg。MS(EI)m/s：1157.5(M+H)。
实施例4、化合物SMP-96745的合成：
Figure PCTCN2021109337-appb-000043
步骤1：目标产物SPM-96745的制备：
50mL反应瓶中依次加入原料4-2(4.4mg),DMF(1.0mL)，HATU(4.4mg)，DIPEA(2mg)室温搅拌反应1分钟。将B原料(20mg)一次性 加到上述的反应液中，室温反应20分钟。HPLC纯化(H 2O：ACN＝10-90％)，得纯品，重10.5mg。MS(EI)m/s：1010.2(M+H)。
实施例5、化合物XIf的合成：
步骤1：中间体X-1的制备：
Figure PCTCN2021109337-appb-000044
25mL反应瓶中依次加入中间体X(27mg),DMF(1.5mL)，HATU(30mg)，DIPEA(31mg)室温搅拌反应5分钟后，将中间体A(30mg)溶于DMF(0.5mL)澄清后缓慢滴加到上述反应液中，室温反应30分钟。点板，中间体A完全反应。HPLC纯化(H 2O：ACN＝10-90％)，得中间体X-1纯品，重18mg。MS(EI)m/s：723.4(M/2+H)。
步骤2：目标产物XIf的制备：
Figure PCTCN2021109337-appb-000045
15mL反应瓶中依次加入原料X-1(15mg),DMF(1.5mL)，HATU(3.9mg)，DIPEA(2.6mg)室温搅拌反应5分钟。将C原料(12mg)溶于DMF(0.5mL)后缓慢滴加到上述的反应液中，室温反应20分钟。HPLC纯化(H 2O：ACN＝10-90％)，得纯品，重4.6mg。MS(EI)m/s：1281.7(M/2+H)。 1H NMR(400MHz,DMSO-d6)δ9.87(s,2H),8.45(d,J＝8.9Hz,1H),7.87(m,2H),7.75(m,2H),7.59–7.43(m,5H),7.43–7.23(m,10H),6.81–6.67(m,3H),6.16(m,2H),5.43–5.26(m,4H),5.23–5.09(m,6H),5.09–4.97(m,3H),4.96–4.87(m,1H),4.43–4.22(m,12H),4.22–4.09(m,4H),4.09–3.70(m,13H),3.64(t,J＝5.9Hz,2H),3.55(m,1H),3.43–3.35(m,1H),3.34–3.20(m,11H),3.19–3.00(m,4H),2.91(d,J＝7.8Hz,6H),2.68(m,1H),2.62–2.24(m,15H),2.22–1.44(m,42H),1.42–1.23(m,4H),1.18(m,3H),1.09(d,J＝7.0Hz,3H),1.02(d,J＝8.0Hz,3H),0.95(d,J＝7.6Hz,3H),0.92–0.78(m,27H).
实施例6、化合物XIb的合成：
Figure PCTCN2021109337-appb-000046
15mL反应瓶中依次加入原料X-1(15mg),DMF(1.5mL)，HATU(3.9mg)，DIPEA(2.6mg)室温搅拌反应5分钟。将B原料(12mg)溶于DMF(0.5mL)后缓慢滴加到上述的反应液中，室温反应20分钟。HPLC纯化(H 2O：ACN＝10-90％)，得纯品，重5.6mg。MS(EI)m/s：1275.5(M/2+H)。
实施例7、化合物SMP-39545的合成：
Figure PCTCN2021109337-appb-000047
步骤1：目标产物SMP-39545的制备：
15mL反应瓶中依次加入原料X(2.3mg),DMF(1.5mL)，HATU(2.3mg)，DIPEA(2mg)室温搅拌反应1分钟。将A原料(15mg)溶于DMF(0.5mL)后缓慢滴加到上述的反应液中，室温反应20分钟。HPLC纯化(H 2O：ACN＝10-90％)，得纯品SMP-39545，重4.5mg。MS(EI)m/s：1272.5(M/2+H)。
实施例8、化合物SMP-93954的合成：
步骤1：中间体X-2的制备：
Figure PCTCN2021109337-appb-000048
25mL反应瓶中依次加入中间体X(45mg),DMF(1.5mL)，HATU(50mg)，DIPEA(22.7mg)室温搅拌反应5分钟后，将中间体A(30mg)溶于DMF(0.5mL)澄清后缓慢滴加到上述反应液中，室温反应30分钟。点板，中间体完全反应。HPLC纯化(H 2O：ACN＝10-90％)，得中间体X-2纯品，重13mg。MS(EI)m/s：729.0(M/2+H)。
步骤2：目标产物SMP-93954的制备:
Figure PCTCN2021109337-appb-000049
15mL反应瓶中依次加入原料X-2(15mg),DMF(1.5mL)，HATU(3.9mg)，DIPEA(2.6mg)室温搅拌反应5分钟。将B原料(8.6mg)溶于DMF(0.5mL)后缓慢滴加到上述的反应液中，室温反应20分钟。HPLC纯化(H 2O：ACN＝10-90％)，得纯品，重9.6mg。MS(EI)m/s：1141.0(M/2+H)。 1H NMR(400MHz,DMSO-d6)δ9.87(s,1H),8.45(d,J＝8.9Hz,1H),8.15–8.02(m, 4H),7.88(m,2H),7.75(d,J＝8.9Hz,1H),7.54–7.46(m,2H),7.39–7.29(m,4H),7.29–7.18(m,6H),6.75(t,J＝7.4Hz,1H),6.70(s,2H),6.16(t,J＝4.8Hz,1H),5.87(s,1H),5.43–5.29(m,2H),5.24–5.17(m,2H),5.17–5.09(m,4H),5.09–4.99(m,4H),4.69(dd,J＝12.3,1.0Hz,1H),4.60–4.49(m,2H),4.42–4.35(m,2H),4.35–4.29(m,3H),4.29–4.21(m,4H),4.21–4.14(m,2H),4.13(m,2H),4.10–3.99(m,4H),3.99–3.86(m,3H),3.83–3.68(m,8H),3.64(m,3H),3.40–3.18(m,5H),3.18–3.01(m,2H),3.01–2.76(m,4H),2.61–2.26(m,13H),2.23(s,3H),2.20–2.09(m,3H),2.09–1.96(m,4H),1.96–1.80(m,7H),1.80–1.63(m,10H),1.63–1.45(m,9H),1.43–1.31(m,2H),1.02(d,J＝8.0Hz,3H),0.96(t,J＝8.1Hz,3H),0.85(t,J＝1.6Hz,3H),0.84(t,J＝1.6Hz,3H).
实施例9、化合物SMP-11115的合成：
Figure PCTCN2021109337-appb-000050
步骤1：目标产物SMP-11115的制备：
15mL反应瓶中依次加入原料X(2.3mg),DMF(1.5mL)，HATU(2.3mg)，DIPEA(2mg)室温搅拌反应1分钟。将C原料(25mg)溶于DMF(0.5mL)后缓慢滴加到上述的反应液中，室温反应20分钟。HPLC纯化(H 2O：ACN＝10-90％)，得纯品SMP-11115，重18mg。MS(EI)m/s：1287.5(M/2+H)。
实施例10、化合物SMP-04404的合成：
Figure PCTCN2021109337-appb-000051
步骤1：目标产物SMP-04404的制备：
15mL反应瓶中依次加入原料X(2.0mg),DMF(1.5mL)，HATU(13mg)，DIPEA(5.3mg)室温搅拌反应1分钟。将B原料(10mg)溶于DMF(0.5mL)后缓慢滴加到上述的反应液中，室温反应20分钟。HPLC纯化(H 2O：ACN＝10-90％)，得纯品SMP-04404，重3.4mg。MS(EI)m/s：1493(M/2+H)。
实施例11、化合物SMP-37241的合成：
步骤1：中间体Y的制备：
Figure PCTCN2021109337-appb-000052
50mL反应瓶中首先加入H 2O:THF＝2:1＝18mL，后依次加入NaHCO 3(148mg)和Y(200mg)，室温搅拌直到溶液澄清后，将L-3(460mg)溶于DME(6mL)中一次性加入到上述反应液，室温反应12h。后处理：向反应液中加入EA:MeOH＝30:1＝25*3(mL)萃取三次，有机相项用无水硫酸钠干燥后减压蒸馏得到L-3-1的粗品。用HPLC纯化(H 2O：ACN＝10-90％)，得中间体L-3-1纯品，重45mg。MS(EI)m/s：327.1(M+H)
步骤2：中间体Y-1的制备：
Figure PCTCN2021109337-appb-000053
25mL反应瓶中依次加入中间体Y(51mg),DMF(2.5mL)，HATU(60.4mg)，DIPEA(23mg)室温搅拌反应5分钟后，将中间体C原料(60mg)溶于DMF(1.0mL)澄清后缓慢滴加到上述反应液中，室温反应30分钟。点板，中间体完全反应。HPLC纯化(H 2O：ACN＝10-90％)，得中间体Y-1纯品，重40mg。MS(EI)m/s：1443.1(M+H)。 1H NMR(400MHz,DMSO-d6)δ9.87(s,1H),8.59(d,J＝8.9Hz,1H),7.76(m,2H),7.54–7.45(m,2H),7.34(m,2H),6.78–6.68(m,3H),6.16(t,J＝4.8Hz,1H),5.41–5.30(m,2H),5.19(m,1H),5.14(m,3H),5.04(m,2H),4.49–4.36(m,2H),4.36–4.29(m,3H),4.29–4.21(m,4H),4.21–4.13(m,2H),4.10–3.99(m,4H),3.96–3.86(m,2H),3.83–3.69(m,2H),3.64(m,3H),3.32(s,3H),3.27(m,2H),3.16–3.02(m,2H),2.71–2.57(m,2H),2.57–2.27(m,9H),2.21–2.14(m,2H),2.11–1.95(m,4H),1.93–1.45(m,24H),1.36(m,2H),1.02(d,J＝8.0Hz,3H),0.85(t,J＝1.6Hz,3H),0.84(t,J＝1.6Hz,3H).
步骤3：目标产物SMP-37241的制备：
Figure PCTCN2021109337-appb-000054
25mL反应瓶中依次加入中间体Y-1(10mg),DMF(1.5mL)，HATU(3.1mg)，DIPEA(1.3mg)室温搅拌反应1分钟后，将中间体B原料(5.8mg)溶于DMF(0.5mL)澄清后缓慢滴加到上述反应液中，室温反应30分钟。点板，中间体完全反应。HPLC纯化(H 2O：ACN＝10-90％)，得目标产物SMP-37241纯品，重2.1mg。MS(EI)m/s：1133.5(M/2+H)。 1H NMR(400MHz,DMSO-d6)δ9.87(s,1H),8.60(d,J＝8.7Hz,1H),8.19–7.99(m,4H),7.90(d,J＝9.1Hz,1H),7.77(m,2H),7.54–7.46(m,2H),7.41–7.29(m,4H),7.30–7.17(m,6H),6.75(t,J＝7.4Hz,1H),6.70(s,2H),6.16(t,J＝4.8Hz,1H),5.87(s,1H),5.41–5.29(m,2H),5.24–5.17(m,2H),5.17–5.10(m,4H),5.09–5.00(m,4H),4.69(dd,J＝12.3,1.0Hz,1H),4.60–4.50(m,3H),4.42–4.36(m,2H),4.36–4.29(m,4H),4.29–4.21(m,4H),4.21–4.14(m,2H),4.13(m,2H),4.10–3.97(m,4H),3.97–3.86(m,2H),3.83–3.68(m,8H),3.64(m,3H),3.36–3.20(m,5H),3.16–3.01(m,2H),2.99–2.78(m,4H),2.61–2.44(m,6H),2.43–2.27(m,5H),2.26–2.10(m,6H),2.09–1.95(m,4H),1.95–1.44(m,27H),1.36(m,2H),1.02(d,J＝8.0Hz,3H),0.96(t,J＝8.1Hz,3H),0.85(t,J＝1.6Hz,3H),0.84(t,J＝1.6Hz,3H).
实施例12、化合物SMP-13432的合成：
Figure PCTCN2021109337-appb-000055
步骤1：目标产物SMP-13432的制备：
25mL反应瓶中依次加入中间体Y-1(10mg),DMF(1.5mL)，HATU(3.6mg)，DIPEA(1.2mg)室温搅拌反应1分钟后，将中间体A原料(7.2mg)溶于DMF(0.5mL)澄清后缓慢滴加到上述反应液中，室温反应30分钟。点板，中间体完全反应。HPLC纯化(H 2O：ACN＝10-90％)，得目标产物SMP-13432纯品，重2.6mg。MS(EI)m/s：1274.5(M/2+H)。
实施例13、化合物SMP-84984的合成：
Figure PCTCN2021109337-appb-000056
步骤1：目标产物SMP-84984的制备：
25mL反应瓶中依次加入中间体Y(1.5mg),DMF(1.5mL)，HATU(5.4mg)，DIPEA(2.4mg)室温搅拌反应1分钟后，将中间体B原料(8mg)溶于DMF(0.5mL)澄清后缓慢滴加到上述反应液中，室温反应30分钟。点板，中间体完全反应。HPLC纯化(H 2O：ACN＝10-90％)，得中间体SMP-84984 纯品，重2.5mg。MS(EI)m/s：986.1(M/2+H)。
实施例14、化合物XIa的合成：
Figure PCTCN2021109337-appb-000057
步骤1：中间体Y-A的制备：
25mL反应瓶中依次加入中间体Y(26mg),DMF(2.5mL)，HATU(31mg)，DIPEA(12mg)室温搅拌反应5分钟后，将中间体A原料(30mg)溶于DMF(1.0mL)澄清后缓慢滴加到上述反应液中，室温反应30分钟。点板，中间体完全反应。HPLC纯化(H 2O：ACN＝10-90％)，得中间体Y-A纯品，重20mg。MS(EI)m/s：1431.1
步骤2：目标产物XIa的制备：
Figure PCTCN2021109337-appb-000058
25mL反应瓶中依次加入中间体Y-A(10mg),DMF(1.5mL)，HATU(2.5mg)，DIPEA(1.2mg)室温搅拌反应1分钟后，将中间体B原料(5.8mg)溶于DMF(0.5mL)澄清后缓慢滴加到上述反应液中，室温反应30分钟。点板，中间体完全反应。HPLC纯化(H 2O：ACN＝10-90％)，得目标产物XIa纯品，重2.5mg。MS(EI)m/s：1127.5(M/2+H)。
实施例15、化合物SMP-04140的合成：
步骤1：中间体Z的制备：
Figure PCTCN2021109337-appb-000059
50mL反应瓶中首先加入H 2O:THF＝2:1＝36mL，后依次加入NaHCO 3(1.05g)和SM1(500mg)，室温搅拌直到溶液澄清后，将L-3(1.9g)溶于DME(18mL)中一次性加入到上述反应液，室温反应12h。后处理：向反应液中加入EA:MeOH＝30:1＝35*3(mL)萃取三次，有机相项用无水硫酸钠干燥后减压蒸馏得到Z的粗品。用HPLC纯化(H 2O：ACN＝10-90％)，得中间体Z纯品，重300mg。MS(EI)m/s：313.1(M+H)
步骤2：中间体Z-1的制备：
Figure PCTCN2021109337-appb-000060
25mL反应瓶中依次加入中间体Z(22mg),DMF(1.5mL)，HATU(26.6mg)，DIPEA(9.0mg)室温搅拌反应5分钟后，将中间体C原料(20mg)溶于DMF(1.0mL)澄清后缓慢滴加到上述反应液中，室温反应30分钟。点板，中间体完全反应。HPLC纯化(H 2O：ACN＝10-90％)，得中间体Z-1纯品，重15mg。MS(EI)m/s：715(M/2+H)。
步骤3：目标产物SMP-04140的制备：
Figure PCTCN2021109337-appb-000061
25mL反应瓶中依次加入中间体Z-1(10mg),DMF(1.5mL)，HATU(4mg)，DIPEA(1.8mg)室温搅拌反应1分钟后，将中间体B原料(5.8mg)溶于DMF(0.5mL)澄清后缓慢滴加到上述反应液中，室温反应30分钟。点板，中间体完全反应。HPLC纯化(H 2O：ACN＝10-90％)，得目标产物SMP-04140纯品，重1.7mg。MS(EI)m/s：1126.5(M/2+H)。
实施例16、化合物SMP-76794的合成：
Figure PCTCN2021109337-appb-000062
步骤1：目标产物SMP-76794的制备：
25mL反应瓶中依次加入中间体Z-1(10mg),DMF(1.5mL)，HATU(4mg)，DIPEA(1.8mg)室温搅拌反应1分钟后，将中间体A原料(8.1mg)溶于DMF(0.5mL)澄清后缓慢滴加到上述反应液中，室温反应30分钟。点板，中间体完全反应。HPLC纯化(H 2O：ACN＝10-90％)，得目标产物SMP-76794纯品，重1.8mg。MS(EI)m/s：1126.5(M/2+H)。
实施例17、化合物SMP-99767的合成：
Figure PCTCN2021109337-appb-000063
步骤1：目标产物SMP-99767的制备：
25mL反应瓶中依次加入中间体Z(1.5mg),DMF(1.5mL)，HATU(5.4mg)，DIPEA(2.5mg)室温搅拌反应1分钟后，将中间体B原料(8mg)溶于DMF(1.0mL)澄清后缓慢滴加到上述反应液中，室温反应30分钟。点板，中间体完全反应。HPLC纯化(H 2O：ACN＝10-90％)，得目标产物SMP-99767纯品，重3mg。MS(EI)m/s：979.3(M/2+H)。
实施例18、化合物SMP-86145的合成：
步骤1：中间体Z-A的制备：
Figure PCTCN2021109337-appb-000064
25mL反应瓶中依次加入中间体Z(21mg),DMF(1.5mL)，HATU(25mg)，DIPEA(8.6mg)室温搅拌反应5分钟后，将中间体A原料(25mg)溶于DMF(1.0mL)澄清后缓慢滴加到上述反应液中，室温反应30分钟。点板，中间体完全反应。HPLC纯化(H 2O：ACN＝10-90％)，得中间体Z-A纯品，重20mg。MS(EI)m/s：708(M/2+H)。
步骤2：目标产物SMP-86145的制备：
Figure PCTCN2021109337-appb-000065
25mL反应瓶中依次加入中间体Z-A(10mg),DMF(1.5mL)，HATU(4.0mg)，DIPEA(1.8mg)室温搅拌反应1分钟后，将中间体B原料(5.9mg)溶于DMF(0.5mL)澄清后缓慢滴加到上述反应液中，室温反应30分钟。点板，中间体完全反应。HPLC纯化(H 2O：ACN＝10-90％)，得目标产物SMP-86145纯品，重1.8mg。MS(EI)m/s：1120(M/2+H)。
实施例19、化合物SMP-54418的合成：
步骤1：中间体V-2的制备：
Figure PCTCN2021109337-appb-000066
25mL反应瓶中依次加入中间体V-3(1g),DMF(20mL)，K2CO3(1.16g)，溴乙酸(593mg)，氮气保护，室温下反应12h。后处理：在55℃下减压蒸馏DMF后，将剩余固体溶于H2O(10mL)中，在0℃下调PH＝3,后用乙酸乙酯和饱和食盐水萃取三到五次，有机相用无水硫酸钠干燥后减压蒸馏得到V-2的粗品。HPLC纯化(H 2O：ACN＝10-90％)，得中间体V-2纯品，重400mg。MS(EI)m/s：340(M+H)。
步骤2：中间体V-1的制备：
Figure PCTCN2021109337-appb-000067
25mL反应瓶中加入中间体V-2(400mg),TFA，氮气保护，室温下反应20分钟。在55℃下减压蒸馏后得到V-1的粗品。HPLC纯化(H 2O：ACN＝10-90％)，得中间体V-2纯品，重140mg。MS(EI)m/s：240(M+H)。
步骤3：中间体V的制备：
Figure PCTCN2021109337-appb-000068
15mL反应瓶中加入原料L-2(123mg),DCM(2mL)，在0℃下缓慢滴加草酰氯(0.1mL)室温搅拌反应30分钟。减压浓缩除去DCM，残余物加入DCM(20mL)；将V-1(140mg)溶于DCM(2mL)中后加入DIPEA(140mg)，在0℃下缓慢滴加上述的DCM溶液，室温搅拌反应2小时。HPLC纯化(H 2O：ACN＝10-90％)，得V纯品，重50mg。MS(EI)m/s：433(M+H)。
步骤4：中间体065-1的制备：
Figure PCTCN2021109337-appb-000069
25mL反应瓶中依次加入中间体V(30mg),DMF(1.5mL)，HATU(25mg)，DIPEA(8.6mg)室温搅拌反应5分钟后，将中间体A(21mg)溶于DMF(0.5mL)澄清后缓慢滴加到上述反应液中，室温反应30分钟。点板，中间体A完全反应。HPLC纯化(H 2O：ACN＝10-90％)，得中间体065-1纯品，重8mg。MS(EI)m/s：769(M/2+H)。
步骤5：目标产物SMP-54418的制备：
Figure PCTCN2021109337-appb-000070
25mL反应瓶中依次加入中间体065-1(8mg),DMF(1.5mL)，DIPEA(1.3mg)，HATU(2.9mg)后一次性加入B原料(4.3mg)室温反应20分钟。点板，中间体完全反应。HPLC纯化(H 2O：ACN＝10-90％)，得目标产物SMP-54418纯品，重7mg。MS(EI)m/s：1181(M/2+H)。
实施例20、化合物SMP-44708的合成：
步骤1：中间体066-1的制备：
Figure PCTCN2021109337-appb-000071
25mL反应瓶中依次加入中间体V(15mg),DMF(1.5mL)，HATU(16mg)，DIPEA(6.9mg)室温搅拌反应5分钟后，将中间体B(30mg)溶于DMF(0.5mL)澄清后缓慢滴加到上述反应液中，室温反应30分钟。点板，中间体B完全反应。HPLC纯化(H 2O：ACN＝10-90％)，得中间体066-1纯品，重8.4mg。MS(EI)m/s：1255.5(M+H)。
步骤2：目标产物SMP-44708的制备：
Figure PCTCN2021109337-appb-000072
25mL反应瓶中依次加入中间体066-1(8.4mg),DMF(1.5mL)，DIPEA(1.2mg)，HATU(3.0mg)后一次性加入C原料(7.5mg)室温反应20分钟。点板，中间体完全反应。HPLC纯化(H 2O：ACN＝10-90％)，得目标产物SMP-44708纯品，重7mg。MS(EI)m/s：1185.5(M/2+H)。 1H NMR(400MHz,DMSO-d6)δ9.87(s,1H),8.15–8.02(m,4H),8.01–7.87(m,3H),7.75(d,J＝8.9Hz,1H),7.53–7.46(m,2H),7.38–7.29(m,4H),7.29–7.18(m,6H),7.12(dt,J＝8.8,1.0Hz,2H),6.89–6.84(m,2H),6.75(t,J＝7.4Hz,1H),6.70(s,2H),6.16(t,J＝4.8Hz,1H),5.87(s,1H),5.42–5.30(m,2H),5.24–5.17(m,2H),5.17–5.09(m,4H),5.09–4.99(m,4H),4.69(m,1H),4.61–4.51(m,2H),4.49(m,2H),4.43–3.98(m,18H),3.96–3.85(m,2H),3.83–3.70(m,8H),3.64(m,3H),3.36–3.19(m,5H),3.19–3.02(m,2H),3.02–2.79(m,6H),2.58–1.96(m,19H),1.96–1.43(m,28H),1.36(m,2H),1.02(d,J＝8.0Hz,3H),0.96(t,J＝8.1Hz,3H),0.85(t,J＝1.6Hz,3H),0.84(t,J＝1.6Hz,3H).
实施例21、化合物XId的合成：
Figure PCTCN2021109337-appb-000073
25mL反应瓶中依次加入实施例1中间体4-3(12mg),DMF(1.5mL)，DIPEA(1.5mg)，HATU(3.5mg)后一次性加入D原料(8.0mg)室温反应20分钟。点板，中间体完全反应。HPLC纯化(H 2O：ACN＝10-90％)，得目标产物XId纯品，重7.3mg。MS(EI)m/s：1299.9(M/2+H)。
实施例22、化合物XIg的合成：
Figure PCTCN2021109337-appb-000074
25mL反应瓶中依次加入实施例5中间体x-1(8.0mg),DMF(1.5mL)，DIPEA(1.0mg)，HATU(3.0mg)后一次性加入D原料(7.0mg)室温反应 20分钟。点板，中间体完全反应。HPLC纯化(H 2O：ACN＝10-90％)，得目标产物XId纯品，重6.7mg。MS(EI)m/s：1282.5(M/2+H)。 1H NMR(400MHz,DMSO-d6)δ9.87(m,2H),8.45(d,J＝8.9Hz,1H),7.87(m,2H),7.73(m,3H),7.57–7.46(m,5H),7.41–7.27(m,11H),7.27–7.15(m,5H),6.70(s,2H),6.16(m,2H),5.42–5.29(m,4H),5.11(m,4H),5.04(m,1H),4.95–4.86(m,1H),4.42–4.20(m,9H),4.18–4.08(m,4H),4.01–3.79(m,6H),3.64(t,J＝5.9Hz,2H),3.55(m,2H),3.44–3.36(m,2H),3.24(s,12H),3.18–3.01(m,4H),3.01–2.86(m,14H),2.65(m,2H),2.58–2.46(m,4H),2.40(s,4H),2.20–1.63(m,24H),1.62–1.44(m,6H),1.42–1.22(m,6H),1.18(m,3H),1.09(m,6H),0.95(d,J＝7.6Hz,6H),0.91–0.79(m,42H).
实施例23是本发明抗体-药物偶联物(ADC)的具体制备实施例。
实施例23、ADC的制备
分别以实施例1～22制得的双药物链接组装单元与抗体为原料，制备对应的ADC。其通用方法如下：
步骤1：抗体还原
将曲妥珠单抗用PBS6.0/EDTA配制成10mg/mL的溶液(3.0mL)，向其中加入添加10mM三(2-羧基乙基)膦盐酸盐(TCEP)水溶液(0.0934mL)及1M磷酸氢二钾水溶液(0.150mL)，确保反应溶液pH在7.3-7.5之间，搅拌1分钟后在37℃孵育1小时。
步骤2：抗体与药物偶联
将上述溶液温度降至10℃，分别添加含有10mM实施例1～22中的一种双药物链接组装单元的DMSO溶液(0.0374mL)，搅拌1分钟后保温静置2小时。
步骤3：纯化
使用10KD的半透膜透析步骤2得到的反应液，透析温度25℃，透析溶剂4L/次，透析时间4小时/次，一共透析4次，所得透析液使用10KD超滤离心管离心浓缩至ADC浓度约5mg/mL，于-20℃～-30℃保存备用。
将制得的ADC分别命名为实施例1-ADC～实施例22-ADC，结构分别如下：
1、实施例14-ADC：
Figure PCTCN2021109337-appb-000075
2、实施例6-ADC：
Figure PCTCN2021109337-appb-000076
3、实施例2-ADC：
Figure PCTCN2021109337-appb-000077
4、实施例21-ADC：
Figure PCTCN2021109337-appb-000078
5、实施例1-ADC：
Figure PCTCN2021109337-appb-000079
6、实施例5-ADC：
Figure PCTCN2021109337-appb-000080
7、实施例22-ADC：
Figure PCTCN2021109337-appb-000081
8、实施例3-ADC：
Figure PCTCN2021109337-appb-000082
Figure PCTCN2021109337-appb-000083
9、实施例4-ADC：
Figure PCTCN2021109337-appb-000084
10、实施例7-ADC：
Figure PCTCN2021109337-appb-000085
11、实施例8-ADC：
Figure PCTCN2021109337-appb-000086
12、实施例9-ADC：
Figure PCTCN2021109337-appb-000087
13、实施例10-ADC：
Figure PCTCN2021109337-appb-000088
14、实施例11-ADC：
Figure PCTCN2021109337-appb-000089
15、实施例12-ADC：
Figure PCTCN2021109337-appb-000090
16、实施例13-ADC：
Figure PCTCN2021109337-appb-000091
17、实施例15-ADC：
Figure PCTCN2021109337-appb-000092
18、实施例16-ADC：
Figure PCTCN2021109337-appb-000093
19、实施例17-ADC：
Figure PCTCN2021109337-appb-000094
20、实施例18-ADC：
Figure PCTCN2021109337-appb-000095
21、实施例19-ADC：
Figure PCTCN2021109337-appb-000096
22、实施例20-ADC：
Figure PCTCN2021109337-appb-000097
实施例24、本发明ADC的DAR值测定
步骤1：ADC还原
用pH8.0、50mmol/L的Tris缓冲液将实施例23制得的各ADC样品分别稀释至1mg/ml，然后加入新鲜制备的DTT储备液(1mol/L)，使二硫苏糖醇(DTT)终浓度为50mmol/L，37℃孵育20～30min，进行还原反应。
步骤2：DAR值测定
将步骤1所得样品注入LC-MS检测，经过反卷积处理后得到L+0、L+1、H+0、H+1、H+2、H+3、H+4的HPLC峰面积，通过计算得到DAR值。
此处DAR值表示待测ADC样品中，一个抗体上所偶联的双药物链接组装单元的平均个数。
结果如下表1所示。
表1 ADC的DAR值测定结果
受试药物 DAR值
实施例3-ADC 1.93
实施例5-ADC 2.31
实施例9-ADC 7.71
实施例7-ADC 5.24
实施例6-ADC 2.03
实施例8-ADC 4.32
实施例10-ADC 4.67
实施例11-ADC 6.39
实施例14-ADC 6.41
实施例17-ADC 7.14
实施例13-ADC 5.69
实施例1-ADC 3.35
实施例2-ADC 4.38
实施例4-ADC 4.27
实施例12-ADC 4.26
实施例15-ADC 2.47
实施例16-ADC 2.58
实施例18-ADC 6.45
实施例19-ADC 7.26
实施例20-ADC 2.07
实施例21-ADC 1.99
实施例22-ADC 3.47
以下通过试验例证明本发明的有益效果。
试验例1：细胞实验操作步骤
(1)试验方法
以每孔3000个SK-BR-3细胞(人乳腺腺癌细胞)和每孔1500个N87细胞(人胃癌细胞)的密度分别铺入96孔细胞板中，培养过夜。
第二天配制各待测ADC(即实施例1-ADC～实施例22-ADC)及阳性对照Trastuzumab-GGFG-Dxd(DS-8201a，一种已知的ADC)，按1:5比例梯度稀释，配制成加药终浓度以500nM为最高浓度的9个浓度梯度点。阳性对照Cisplatin(顺铂)按1:3比例梯度稀释，配制成加药终浓度为以100μM为最高浓度的9个浓度梯度点。
将配制好的各待测ADC及阳性对照加入细胞中，以只加入培养基的孔作为阴性对照。加药后培养144小时，然后用CTG(CELL TITER-GLO)法检测细胞活力，分别计算出各受试药物对SK-BR-3细胞和N87细胞的IC50值。结果如下表2所示。
(2)试验结果
表2 各药物对SK-BR-3细胞和N87细胞的IC 50值
Figure PCTCN2021109337-appb-000098
Figure PCTCN2021109337-appb-000099
上述实验结果表明，与化疗药物Cisplatin(顺铂)相比，本发明提供的ADC能够显著提高对乳腺腺癌细胞和胃癌细胞的抑制作用。与已经上市的ADC：DS-8201a相比，本发明提供的ADC对乳腺腺癌细胞和胃癌细胞的抑制作用也都明显提高。
综上，本发明提供了一种双药物链接组装单元，该双药物链接组装单元能够与靶向接头连接得到对应的双药靶向接头-药物偶联物，该双药靶向接头-药物偶联物能够靶向作用于肿瘤细胞，降低对正常细胞的毒副作用，同时能够有效克服药物的耐药性。与已经上市的ADC：DS-8201a相比，本发明提供的ADC对乳腺腺癌细胞和胃癌细胞的抑制作用都明显提高。本发明的双药物链接组装单元和双药靶向接头-药物偶联物在制备预防和/或治疗的抗肿瘤药物中具有广阔的应用前景。

Claims (19)

  1. 式VI所示的双药物链接组装单元、或其立体异构体、或其旋光异构体：
    Figure PCTCN2021109337-appb-100001
    其中，T是拴系基团，它能够与靶向接头连接；所述靶向接头为能够靶向结合病变部位的物质；
    U是三叉型连接子部分；
    L1和L2为可断裂或不可断裂的链接基团；
    D 1、D 2分别是第一药物单元和第二药物单元。
  2. 根据权利要求1所述的双药物链接组装单元、或其立体异构体、或其旋光异构体，其特征在于：所述T能够与靶向接头上的巯基反应连接。
  3. 根据权利要求1或2所述的双药物链接组装单元、或其立体异构体、或其旋光异构体，其特征在于：所述双药物链接组装单元的结构如式VI-1所示：
    Figure PCTCN2021109337-appb-100002
    其中，K 1、K 2各自独立的选自无、K aK bK c；K a为苯基，K b选自O、S或无，K c选自0～2个亚甲基；
    n，p各自独立的选自0～10的整数；
    U为取代或未取代的芳基、杂芳基、链烷基、环烷基或杂环烷基；所述取代基选自氘、卤素、氰基、羟基、C 1～6烷基或C 1～6烷氧基；
    X 1、X 2各自独立的选自
    Figure PCTCN2021109337-appb-100003
    其中，a、b、c、d各自独立的选自0或1，K 3选自无、无取代或被R 1取代的C 1～4亚烷基，K 4选自无、无取代或被R 2取代的C 1～4亚烷基，K 5选自无、无取代或被R 3取代的C 1～4亚烷基，K 6选自无、无取代或被R 4取代的C 1～4亚烷基，R 1、R 2、 R 3、R 4各自独立的选自H、C 1～5烷基、取代或者未取代的苄基、-L 1aNHCONH 2、-L 1aNHCOCR 5NH 2，L 1a为0～4个亚甲基，R 5为C 1～3烷基；所述苄基上的取代基选自氘、卤素、氰基、羟基、C 1～6烷基或C 1～6烷氧基；
    B 1，B 2，C 1，C 2，E 1，E 2各自独立的选自取代或未取代的以下基团：
    Figure PCTCN2021109337-appb-100004
    L 2aCONHL 3、L 4OL 5CO或无；所述取代基各自独立的选自
    Figure PCTCN2021109337-appb-100005
    C 1～5烷基；其中L 2a、L 3、L 4、L 5、L 6各自独立的选自0～2个亚甲基；
    D 1、D 2各自独立的选自细胞毒性药物、治疗自身免疫疾病的药物或抗炎的药物；
    T如权利要求1或2所述。
  4. 根据权利要求3所述的双药物链接组装单元、或其立体异构体、或其旋光异构体，其特征在于：所述双药物链接组装单元的结构如式VII所示：
    Figure PCTCN2021109337-appb-100006
    其中，n，p各自独立的选自0～10的整数；
    U为取代或未取代的芳基、杂芳基、链烷基、环烷基或杂环烷基；所述取代基选自氘、卤素、氰基、羟基、C 1～6烷基或C 1～6烷氧基；
    X 1、X 2各自独立的选自
    Figure PCTCN2021109337-appb-100007
    其中，a、b、c、d各自独立的选自0或1，R 1、R 2、R 3、R 4各自独立的选自H、C 1～5烷基、取代或者未取代的苄基、-L 1aNHCONH 2，L 1a为0～3个亚甲基；所述苄基上的取代基选自氘、卤素、氰基、羟基、C 1～6烷基或C 1～6烷氧基；
    B 1，B 2，C 1，C 2，E 1，E 2各自独立的选自取代或未取代的以下基团：
    Figure PCTCN2021109337-appb-100008
    L 2aCONHL 3、L 4OL 5CO或无；所述取代基各自独立的选自
    Figure PCTCN2021109337-appb-100009
    C 1～5烷基；其中L 2a、L 3、L 4、L 5、L 6各自独立的选自0～2个亚甲基；
    D 1、D 2各自独立的选自细胞毒性药物、治疗自身免疫疾病的药物或抗炎的药物；
    T如权利要求3所述。
  5. 根据权利要求4所述的双药物链接组装单元、或其立体异构体、或其旋光异构体，其特征在于：所述双药物链接组装单元的结构如式VII-1所示：
    Figure PCTCN2021109337-appb-100010
    其中，A 1、A 2各自独立的选自0～10个亚甲基，O，S，CO，NH，CONH，NHCO或无；
    m，n，p，U，X 1，X 2，B 1，B 2，C 1，C 2，E 1，E 2，D 1、D 2如权利要求4所述。
  6. 根据权利要求5所述的双药物链接组装单元、或其立体异构体、或其旋光异构体，其特征在于：所述双药物链接组装单元的结构如式VIIIa或VIIIb所示：
    Figure PCTCN2021109337-appb-100011
    其中，A 1、A 2、m、n、p、X 1、X 2、B 1、B 2、C 1、C 2、E 1、E 2、D 1、D 2如权利要求5所述；
    M为氢、氘、卤素、氰基、羟基、C 1～6烷基或C 1～6烷氧基。
  7. 根据权利要求6所述的双药物链接组装单元、或其立体异构体、或其旋光异构体，其特征在于：所述双药物链接组装单元的结构选自以下结构之一：
    Figure PCTCN2021109337-appb-100012
    Figure PCTCN2021109337-appb-100013
    其中，D 1、D 2如权利要求6所述，n、p为0～4的整数；优选地，n＝p。
  8. 根据权利要求7所述的双药物链接组装单元、或其立体异构体、或其旋光异构体，其特征在于：所述双药物链接组装单元的结构为以下结构之一：
    Figure PCTCN2021109337-appb-100014
    Figure PCTCN2021109337-appb-100015
    Figure PCTCN2021109337-appb-100016
    其中，D 1、D 2如权利要求7所述。
  9. 根据权利要求1～8任一项所述的双药物链接组装单元、或其立体异构体、或其旋光异构体，其特征在于：所述D 1、D 2各自独立的选自以DNA为靶标的药物单元或以微管蛋白为靶标的药物单元；所述以DNA为靶标的药物单元优选为SN-38，Dxd，Dx-8951或其衍生物，和/或，所述以微管蛋白为靶标的药物单元优选为Eribulin，MMAE，MMAF，美登素或其衍生物。
  10. 根据权利要求1～3任一项所述的双药物链接组装单元、或其立体异构体、或其旋光异构体，其特征在于：所述双药物链接组装单元为以下结构之一：
    Figure PCTCN2021109337-appb-100017
    Figure PCTCN2021109337-appb-100018
    Figure PCTCN2021109337-appb-100019
    Figure PCTCN2021109337-appb-100020
    Figure PCTCN2021109337-appb-100021
    Figure PCTCN2021109337-appb-100022
    Figure PCTCN2021109337-appb-100023
    Figure PCTCN2021109337-appb-100024
    Figure PCTCN2021109337-appb-100025
  11. 一种双药靶向接头-药物偶联物分子、或其立体异构体、或其旋光异构体，其特征在于：所述双药靶向接头-药物偶联物分子是由靶向接头和q个权利要求1～10任一项所述的双药物链接组装单元连接所得；所述靶向接头为能够靶向结合病变部位的物质；所述双药靶向接头-药物偶联物分子的结构如式I所示：
    Figure PCTCN2021109337-appb-100026
    其中，Ab为靶向接头，q为1～20的整数，T、U、L1、L2、D 1、D 2如权利要求1～10任一项所述。
  12. 根据权利要求11所述的双药靶向接头-药物偶联物分子、或其立体异构体、或其旋光异构体，其特征在于：所述靶向接头为抗体、抗体片段、蛋白或核酸适配体，所述抗体优选为针对细胞表面受体和肿瘤相关抗原的抗体；
    和/或，所述q为1～8的整数。
  13. 根据权利要求11或12所述的双药靶向接头-药物偶联物分子、或其立体异构体、或其旋光异构体，其特征在于：所述双药靶向接头-药物偶联物分子的结构选自以下结构之一：
    Figure PCTCN2021109337-appb-100027
    Figure PCTCN2021109337-appb-100028
    Figure PCTCN2021109337-appb-100029
    Figure PCTCN2021109337-appb-100030
    Figure PCTCN2021109337-appb-100031
    Figure PCTCN2021109337-appb-100032
    Figure PCTCN2021109337-appb-100033
    Figure PCTCN2021109337-appb-100034
    Figure PCTCN2021109337-appb-100035
    Figure PCTCN2021109337-appb-100036
  14. 一种双药靶向接头-药物偶联物、或其立体异构体、或其旋光异构体，其特征在于：所述双药靶向接头-药物偶联物是由靶向接头和权利要求1～10任一项所述的双药物链接组装单元连接所得；所述靶向接头为能够靶向结合病变部位的物质，优选为抗体、抗体片段、蛋白或核酸适配体；所述抗体优选为针对细胞表面受体和肿瘤相关抗原的抗体。
  15. 根据权利要求14所述的双药靶向接头-药物偶联物、或其立体异构体、或其旋光异构体，其特征在于：所述双药靶向接头-药物偶联物由权利要求13中式IIa所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为6.00～7.00，优选为6.41；
    或，所述双药靶向接头-药物偶联物由权利要求13中式IIb所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为1.50～2.50，优选为2.03；
    或，所述双药靶向接头-药物偶联物由权利要求13中式IIc所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为4.00～5.00，优选为4.38；
    或，所述双药靶向接头-药物偶联物由权利要求13中式IId所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为1.50～2.50，优选为1.99；
    或，所述双药靶向接头-药物偶联物由权利要求13中式IIe所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为3.00～4.00，优选为3.35；
    或，所述双药靶向接头-药物偶联物由权利要求13中式IIf所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联 物的DAR值为2.00～3.00，优选为2.31；
    或，所述双药靶向接头-药物偶联物由权利要求13中式IIg所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为3.00～4.00，优选为3.47；
    或，所述双药靶向接头-药物偶联物由权利要求13中式II SMP-52581所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为1.50～2.50，优选为1.93；
    或，所述双药靶向接头-药物偶联物由权利要求13中式II SMP-96745所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为4.00～5.00，优选为4.27；
    或，所述双药靶向接头-药物偶联物由权利要求13中式II SMP-39545所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为5.00～6.00，优选为5.24；
    或，所述双药靶向接头-药物偶联物由权利要求13中式II SMP-93954所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为4.00～5.00，优选为4.32；
    或，所述双药靶向接头-药物偶联物由权利要求13中式II SMP-11115所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为7.00～8.00，优选为7.71；
    或，所述双药靶向接头-药物偶联物由权利要求13中式II SMP-04404所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为4.00～5.00，优选为4.67；
    或，所述双药靶向接头-药物偶联物由权利要求13中式II SMP-37241所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为6.00～7.00，优选为6.39；
    或，所述双药靶向接头-药物偶联物由权利要求13中式II SMP13432所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为4.00～5.00，优选为4.26；
    或，所述双药靶向接头-药物偶联物由权利要求13中式II SMP-84984所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为5.00～6.00，优选为5.69；
    或，所述双药靶向接头-药物偶联物由权利要求13中式II SMP-04140所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为2.00～3.00，优选为2.47；
    或，所述双药靶向接头-药物偶联物由权利要求13中式II SMP-76794所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为2.00～3.00，优选为2.58；
    或，所述双药靶向接头-药物偶联物由权利要求13中式II SMP-99767所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头 -药物偶联物的DAR值为6.50～7.50，优选为7.14；
    或，所述双药靶向接头-药物偶联物由权利要求13中式II SMP-86145所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为6.00～7.00，优选为6.45；
    或，所述双药靶向接头-药物偶联物由权利要求13中式II SMP-54418所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为7.00～8.00，优选为7.26；
    或，所述双药靶向接头-药物偶联物由权利要求13中式II SMP-44708所示双药靶向接头-药物偶联物分子中的两种或两种以上组成，该双药靶向接头-药物偶联物的DAR值为1.50～2.50，优选为2.07。
  16. 权利要求11～13任一项所述双药靶向接头-药物偶联物分子，或权利要求14～15任一项所述双药靶向接头-药物偶联物的制备方法，其特征在于：所述方法包括以下步骤：将靶向接头与双药物链接组装单元偶联，纯化，即得；优选地，所述纯化方式为透析。
  17. 一种预防和/或治疗肿瘤的药物，其特征在于：它是以权利要求11～13任一项所述的双药靶向接头-药物偶联物分子、或其立体异构体、或其旋光异构体，或权利要求14～15任一项所述双药靶向接头-药物偶联物、或其立体异构体、或其旋光异构体为活性成分，加上药学上可接受的辅料制成的制剂。
  18. 权利要求11～13任一项所述的双药靶向接头-药物偶联物分子、或其立体异构体、或其旋光异构体，或权利要求14～15任一项所述双药靶向接头-药物偶联物、或其立体异构体、或其旋光异构体在制备预防和/或治疗肿瘤的药物中的用途。
  19. 根据权利要求18所述的用途，其特征在于：所述肿瘤选自肺癌、尿道癌、大肠癌、前列腺腺癌、卵巢癌、胰腺癌、乳腺癌、膀胱癌、胃癌、胃肠道间质瘤、宫颈癌、食道癌、鳞状细胞癌、腹膜癌、肝癌、结肠癌、直肠癌、结直肠癌、子宫癌、唾液腺癌、肾癌、外阴癌、甲状腺癌、阴茎癌、白血病、恶性淋巴瘤、浆细胞瘤、骨髓瘤或肉瘤。
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