WO2023124537A1 - Conjugué et conjugué anticorps-médicament préparé à l'aide de celui-ci - Google Patents

Conjugué et conjugué anticorps-médicament préparé à l'aide de celui-ci Download PDF

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WO2023124537A1
WO2023124537A1 PCT/CN2022/129983 CN2022129983W WO2023124537A1 WO 2023124537 A1 WO2023124537 A1 WO 2023124537A1 CN 2022129983 W CN2022129983 W CN 2022129983W WO 2023124537 A1 WO2023124537 A1 WO 2023124537A1
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antibody
conjugate
alkylene
straight chain
compound
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PCT/CN2022/129983
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Chinese (zh)
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郝婧
王庆彬
闫胜勇
冯丛然
郭军
赵宣
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辽宁键凯科技有限公司
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2863—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against receptors for growth factors, growth regulators
    • 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
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/4745—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines
    • 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
    • A61K47/62—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
    • 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
    • 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
    • A61K47/6845—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 cytokine, e.g. growth factors, VEGF, TNF, a lymphokine or an interferon
    • 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
    • A61K47/6889—Conjugates wherein the antibody being the modifying agent and wherein the linker, binder or spacer confers particular properties to the conjugates, e.g. peptidic enzyme-labile linkers or acid-labile linkers, providing for an acid-labile immuno conjugate wherein the drug may be released from its antibody conjugated part in an acidic, e.g. tumoural or environment
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/06034—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/10—Tetrapeptides
    • C07K5/1002—Tetrapeptides with the first amino acid being neutral
    • C07K5/1005—Tetrapeptides with the first amino acid being neutral and aliphatic
    • C07K5/1008—Tetrapeptides with the first amino acid being neutral and aliphatic the side chain containing 0 or 1 carbon atoms, i.e. Gly, Ala

Definitions

  • the present invention relates to the field of medicine, in particular to the field of drug conjugates, in particular to a conjugate and an antibody-drug conjugate prepared therefrom.
  • Antibody drug conjugate is a new type of targeted therapy drug, which is composed of a monoclonal antibody targeting a specific antigen and a small molecule cytotoxic drug coupled through a linker, which combines the antibody Compared with traditional small molecule antineoplastic drugs, it has the characteristics of strong targeting, high cytotoxicity, long degradation half-life, and low toxic and side effects. along with and 14 other drugs have been approved by the FDA. In recent years, ADC drugs have developed rapidly. Currently, about 110 ADC drugs are in clinical trials.
  • ADC generally consists of three parts: antibody, small molecule cytotoxin and linker.
  • the design of the linker is of great significance to the ADC drug, which must keep the drug stable in the blood circulation system and release the active toxin quickly and effectively after reaching the target tissue.
  • linkers there are many important considerations, including the conjugation site of the antibody, the average number of cytotoxins per molecule of antibody (drug to antibody ratio, DAR), the cleavability and hydrophilicity of the linker sex etc.
  • Linkers can be divided into two categories: cleavable linkers and non-cleavable linkers, and cleavable linkers are divided into pH-sensitive and enzyme-sensitive types according to the different cleavage mechanisms.
  • enzyme-sensitive linkers have become the mainstream choice for ADCs.
  • the more mature enzyme-sensitive linker is a dipeptide linker dependent on cathepsin B cleavage, such as Valine-Citriline.
  • Most current ADCs share common structural features, such as linkage via maleimide linkers.
  • the published linkers of ADC drugs have problems such as relatively simple design and poor water solubility.
  • ADC drugs are mostly randomly coupled with lysine or cysteine. Due to the uncertainty of the number and sites of coupling, the generated antibody-drug conjugates are inconsistent. Uniformity, which in turn leads to inhomogeneity in pharmacokinetic properties, potency and toxicity among the components of the product.
  • Patent document CN108743968A discloses a cysteine modified antibody-toxin conjugate, the antibody is a cysteine site-directed insertion antibody, and the cysteine insertion site contains kappa/ ⁇ light chain constant region light chain and IgG anti- There are two types of heavy chains in the constant region of the heavy chain.
  • the antibody-toxin conjugates introduced by genetic engineering introduce uncertainty in the drug, and it is necessary to use a reducing agent to reduce the antibody, release the shield on the modified cysteine residue on the antibody, and pass through cation exchange chromatography or ultrasonography. Removing DTT and shielding materials by means of filtration and replacement, and then using an oxidizing agent to oxidize the antibody to reconnect the interchain disulfide bonds of the antibody, the cumbersome and complicated process limits the application of this invention.
  • Patent document CN103083680B discloses a drug conjugate having a general structural formula of polyethylene glycol-amino acid oligopeptide-irnotecan.
  • each end group of polyethylene glycol can be connected with multiple irinotecans through amino acid oligopeptides, and the drug loading rate is greatly improved.
  • the modification of the hydrophilic polymer can protect inotecan, improve drug absorption, prolong action time, enhance curative effect, reduce dosage and avoid toxic and side effects.
  • the homogeneity of the antibody-drug conjugate is not high, which leads to inhomogeneity in pharmacokinetic properties, potency and toxicity among the components of the product, poor reproducibility of product batches, and low therapeutic index.
  • the first aspect of the present invention is to provide a conjugate 1, which has the structure shown in formula (I):
  • the Q is selected from: one of
  • R 5 and R 6 each independently have the structure of -X 1 -Q 1 , Q 1 is selected from -H, -F, -Cl, -Br, -I, -SO 2 , -NO 2 , C 1-12 chains Alkyl, C 3-12 cycloalkyl, C 6-12 aralkyl, X 1 is selected from single bond, -O-, -S-, C 1-12 chain alkyl, C 3-12 cycloalkyl , C 6-12 Aralkyl, and or a combination thereof;
  • X is a linking group selected from -O-, -S-, C 1-12 straight chain/branched chain alkyl, C 3-12 cycloalkyl, C 6-12 aralkyl, and or a group consisting of combinations thereof, wherein R is selected from: -H, C 1-10 straight chain/branched alkyl;
  • L is selected from: one of straight chain, Y-type and multi-branched polyethylene glycol residues
  • L 1 is a Y-shaped and multi-branched polyethylene glycol residue, it can have one or more branches and connected.
  • L 2 is a linking group, for Wherein A is a peptide linker, and B is selected from: Wherein, Y 1 and Y 2 are each independently selected from: -H, halogen, -OC 1-10 alkyl, C 1-10 straight chain/branched chain alkyl, C 3-10 cycloalkyl, -OH, Preferably, both Y 1 and Y 2 are -H;
  • Y 3 and Y 4 are each independently selected from: -H, C 1-10 straight chain/branched chain alkyl, C 3-10 cycloalkyl, Preferably, both Y 3 and Y 4 are -CH 3 ;
  • B is
  • said B is
  • D is a drug molecule.
  • Q is X is More preferably, R5 and R6 are H.
  • the conjugate 1 has the structure shown in formula (II):
  • the DBCO (dibenzocyclooctyne) linker in the formula (I) is used for site-specific coupling with the azide group of the modified antibody;
  • L1 is selected from: straight chain, Y-type and multi-branched polyethylene glycol
  • L 1 includes but not limited to straight-chain double-ended PEG, Y-type PEG, 4-arm branched-chain PEG, 6-arm branched-chain PEG or 8-arm branched-chain PEG, preferably, L 1 is straight-chain
  • Polyethylene glycol residue has a structure shown in general formula (III):
  • n is independently selected from an integer of 1-30, preferably an integer of 1-25, more preferably an integer of 4-24 (such as 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24).
  • said n1 is 8.
  • n 1 is 24.
  • the peptide linker includes one or more identical or different polypeptide residues, and the polypeptide residues include two or more identical or different amino acid residues or amino acid residue derivatives, and the Amino acids selected from: Alanine, Arginine, Asparagine, Aspartic Acid, Cysteine, Glutamine, Glutamic Acid, Glycine, Histidine, Isoleucine, Leucine, Lys One of amino acid, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, citrulline, ornithine, and cystine or a combination of two or more, preferably, the amino acid is selected from: valine, citrulline, glycine, alanine, phenylalanine, tyrosine, isoleucine, leucine and arginine One or a combination of two or more amino acids, the polypeptide residues include one of valine-citrulline, valine
  • R 1 , R 2 , R 3 , and R 4 are each independently selected from: -H, -C 1-6 straight chain/branched chain alkyl, -(C 1-6 alkylene )-OH, -(C 1-6 alkylene)-SH, -(CH 2 ) 1-6 -cycloalkyl, -(C 1-6 alkylene)-aryl ring, -(C 1- 6 alkylene)-heterocycloalkyl, -(C 1-6 alkylene)-heterocyclic aryl, -(C 1-6 alkylene)-COOH, -(C 1-6 alkylene) -CONH 2 , -(C 1-6 alkylene)-NH 2 and One of them, preferably, said R 1 , R 2 , R 3 , and R 4 are each independently selected from: -H, -C 1-6 straight chain/branched chain alkyl, -(C 1-6 alkylene )-OH, -(C 1-6 al
  • the R 1 is selected from one of: -H, methyl, ethyl, isopropyl, tert-butyl, isobutyl and isopentyl
  • the R 2 is selected from: -H , methyl, ethyl, isopropyl, tert-butyl, isobutyl, isopentyl and One of them
  • the R 3 is (C 1 alkylene)-phenyl, (C 2 alkylene)-phenyl, (C 3 alkylene)-phenyl, (C 4 alkylene )-phenyl, (C 5 alkylene)-phenyl and (C 6 alkylene)-phenyl
  • the R 4 is selected from: -H, methyl, ethyl, isopropyl
  • said R 1 is -H or isopropyl
  • said R 2 is selected from: -
  • said A is selected from:
  • the L2 is selected from: A combination of one or more of, said R 1 , R 2 , R 3 , R 4 are each independently selected from: -H, -C 1-6 straight chain/branched chain alkyl, -(C 1 -6 alkylene)-OH, -(C 1-6 alkylene)-SH, -(CH 2 ) 1-6 -cycloalkyl, -(C 1-6 alkylene)-aryl ring, -(C 1-6 alkylene)-heterocycloalkyl, -(C 1-6 alkylene)-heterocyclic aryl, -(C 1-6 alkylene)-COOH, -(C 1- 6 alkylene) -CONH 2 , -(C 1-6 alkylene) -NH 2 and One of them, preferably, the L2 is selected from: One of.
  • said R 1 is selected from: -H, -C 1-6 straight chain/branched chain alkyl, and one of (C 1-6 alkylene)-aromatic ring groups, more preferably, said R 1 is selected from: -H, methyl, ethyl, isopropyl, tert-butyl, isobutyl One of radical and isopentyl, particularly preferably, said R 1 is -H or isopropyl.
  • said R 2 is selected from: -H, -C 1-6 straight chain/branched chain alkyl, and (C 1-6 alkylene)-aryl ring group, more preferably, the R 2 is selected from: -H, methyl, ethyl, isopropyl, tert-butyl, isobutyl base, isopentyl and One of, particularly preferably, said R 2 is selected from: -H, methyl and One of.
  • said R 3 is (C 1-6 alkylene)-aromatic ring group, more preferably, said R 3 is (C 1 alkylene)-phenyl, (C 2 alkylene )-phenyl, (C 3 alkylene)-phenyl, (C 4 alkylene)-phenyl, (C 5 alkylene)-phenyl and (C 6 alkylene)-phenyl
  • said R 3 is (C 1 alkylene)-phenyl.
  • said R 4 is -H, -C 1-6 straight chain/branched chain alkyl, and one of (C 1-6 alkylene)-aromatic ring groups, more preferably, said R 4 is selected from: -H, methyl, ethyl, isopropyl, tert-butyl, isobutyl One of radical and isopentyl, particularly preferably, said R 4 is -H.
  • said L is selected from: One of.
  • the drug molecule is selected from: amanitin, auristatin, calicheamicin, camptothecin, camptothecin derivatives and metabolites (SN-38), cryptomycin, Dao Normycin, Dolastatin, Doxorubicin, Duocamycin, Epothilone, Espamycin, Geldanamycin, Maytansine, Methotrexate, Monomethylol Restatin E (“MMAE”), monomethyl auristatin F (“MMAF”), pyrrolobenzodiazepine, rhizobin, SG2285, tubulysin, vindesine, toxoid, or any of the above derivative;
  • amanitin auristatin
  • calicheamicin calicheamicin
  • camptothecin camptothecin derivatives and metabolites
  • SN-38 metabolites
  • cryptomycin Dao Normycin
  • Dolastatin Doxorubicin
  • Duocamycin Epothilone
  • Espamycin
  • the drug molecule is selected from one of: SN38 and its derivatives, Dxd, exitecan, MMAE, MMAF, MMAD, PBD and its derivatives, calicheamicin and TPL.
  • the drug molecule is selected from: One of them, preferably, the drug molecule is selected from:
  • the DBCO linker-drug conjugate is selected from:
  • the second aspect of the present invention is to provide a preparation method of the conjugate 1 as described in the general formula (II), the preparation method comprising the following steps:
  • step (1) Compound 1 obtained in step (1) is synthesized by reaction with N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide and denote it as compound 2;
  • step (3) Under the action of the reagent 4-dimethylaminopyridine, compound 2 obtained in step (2) is reacted with H 2 NL 3 -OH to synthesize and denote it as compound 3;
  • R 7 is selected from one of: -H, or C 1-10 straight chain/branched chain alkyl, preferably, said R 7 is C 1-5 straight chain/branched chain alkyl, more preferably, The R 7 is -CH 3 ;
  • the R 8 is Wherein Y 5 is C 1-10 straight chain/branched chain alkyl, preferably, said Y 5 is C 1-5 straight chain/branched chain alkyl, more preferably, said Y 5 is -CH 2 -CH2- ;
  • Y is selected from: -H, C 1-10 straight chain/branched chain alkyl, C 3-10 cycloalkyl, Preferably, said Y 4 is a C 1-5 straight chain/branched chain alkyl group, more preferably, said Y 4 is -CH 3 .
  • the DBCO linker-drug conjugate of the present invention can be used in the preparation of antibody drug conjugates.
  • the third aspect of the present invention is to provide a conjugate 2, which has the structure shown in formula (VI):
  • the conjugate 2 has the structure shown in formula (VII):
  • the conjugate 2 is selected from the structures shown in formulas (VII-1) and (VII-2):
  • the conjugate 2 is selected from:
  • n is selected from an integer of 1-30 (such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30), preferably, n is an integer selected from 2-15.
  • n 4.
  • n 12.
  • n is independently selected from an integer of 1-30, preferably an integer of 1-25, more preferably an integer of 4-24 (such as 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24).
  • said n1 is 8.
  • n 1 is 24.
  • d is an integer selected from 1-10 (such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10), preferably, d is an integer selected from 1-5;
  • d is 1.
  • d is 2.
  • L1 is selected from: one of straight chain, Y-type and multi-branched polyethylene glycol residues, L1 includes but not limited to straight-chain double-ended PEG, Y-type PEG, 4-arm branched chain PEG, 6-arm branched Chain PEG or 8-arm branched chain PEG, when L 1 is a Y-type and multi-branched polyethylene glycol residue, it can have one or more branches and connected.
  • L is a straight-chain polyethylene glycol residue with a structure shown in general formula (III):
  • n is independently selected from an integer of 1-30, preferably an integer of 1-25, more preferably an integer of 4-24 (such as 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24).
  • said n1 is 8.
  • n 1 is 24.
  • L 2 is a linking group, for Wherein A is a peptide linker, and B is selected from: Wherein, Y 1 and Y 2 are each independently selected from: -H, halogen, -OC 1-10 alkyl, C 1-10 straight chain/branched chain alkyl, C 3-10 cycloalkyl, -OH, Preferably, both Y 1 and Y 2 are -H;
  • Y 3 and Y 4 are each independently selected from: -H, C 1-10 straight chain/branched chain alkyl, C 3-10 cycloalkyl, Preferably, both Y 3 and Y 4 are -CH 3 ;
  • B is
  • said B is
  • D is a drug molecule.
  • the peptide linker includes one or more identical or different polypeptide residues
  • the polypeptide residues include two or more identical or different amino acid residues or amino acid residue derivatives
  • Described amino acid is selected from: alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine acid, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, citrulline, ornithine, and cystine
  • the amino acid is selected from the group consisting of: valine, citrulline, glycine, alanine, phenylalanine, tyrosine, isoleucine, leucine acid and arginine or a combination of two or more
  • the polypeptide residues include valine-citrulline, valine-alanine and
  • R 1 , R 2 , R 3 , and R 4 are each independently selected from: -H, -C 1-6 straight chain/branched chain alkyl, -(C 1-6 alkylene )-OH, -(C 1-6 alkylene)-SH, -(CH 2 ) 1-6 -cycloalkyl, -(C 1-6 alkylene)-aryl ring, -(C 1- 6 alkylene)-heterocycloalkyl, -(C 1-6 alkylene)-heterocyclic aryl, -(C 1-6 alkylene)-COOH, -(C 1-6 alkylene) -CONH 2 , -(C 1-6 alkylene)-NH 2 and One of them, preferably, said R 1 , R 2 , R 3 , and R 4 are each independently selected from: -H, -C 1-6 straight chain/branched chain alkyl, -(C 1-6 alkylene )-OH, -(C 1-6 al
  • the R 1 is selected from one of: -H, methyl, ethyl, isopropyl, tert-butyl, isobutyl and isopentyl
  • the R 2 is selected from: -H , methyl, ethyl, isopropyl, tert-butyl, isobutyl, isopentyl and One of them
  • the R 3 is (C 1 alkylene)-phenyl, (C 2 alkylene)-phenyl, (C 3 alkylene)-phenyl, (C 4 alkylene )-phenyl, (C 5 alkylene)-phenyl and (C 6 alkylene)-phenyl
  • the R 4 is selected from: -H, methyl, ethyl, isopropyl
  • said R 1 is -H or isopropyl
  • said R 2 is selected from: -
  • said A is selected from:
  • the L2 is selected from: A combination of one or more of, said R 1 , R 2 , R 3 , R 4 are each independently selected from: -H, -C 1-6 straight chain/branched chain alkyl, -(C 1 -6 alkylene)-OH, -(C 1-6 alkylene)-SH, -(CH 2 ) 1-6 -cycloalkyl, -(C 1-6 alkylene)-aryl ring, -(C 1-6 alkylene)-heterocycloalkyl, -(C 1-6 alkylene)-heterocyclic aryl, -(C 1-6 alkylene)-COOH, -(C 1- 6 alkylene) -CONH 2 , -(C 1-6 alkylene) -NH 2 and One of them, preferably, the L2 is selected from: One of.
  • said R 1 is selected from: -H, -C 1-6 straight chain/branched chain alkyl, and one of (C 1-6 alkylene)-aromatic ring groups, more preferably, said R 1 is selected from: -H, methyl, ethyl, isopropyl, tert-butyl, isobutyl One of radical and isopentyl, particularly preferably, said R 1 is -H or isopropyl.
  • said R 2 is selected from: -H, -C 1-6 straight chain/branched chain alkyl, and (C 1-6 alkylene)-aryl ring group, more preferably, the R 2 is selected from: -H, methyl, ethyl, isopropyl, tert-butyl, isobutyl base, isopentyl and One of, particularly preferably, said R 2 is selected from: -H, methyl and One of.
  • said R 3 is (C 1-6 alkylene)-aromatic ring group, more preferably, said R 3 is (C 1 alkylene)-phenyl, (C 2 alkylene )-phenyl, (C 3 alkylene)-phenyl, (C 4 alkylene)-phenyl, (C 5 alkylene)-phenyl and (C 6 alkylene)-phenyl
  • said R 3 is (C 1 alkylene)-phenyl.
  • said R 4 is -H, -C 1-6 straight chain/branched chain alkyl, and one of (C 1-6 alkylene)-aromatic ring groups, more preferably, said R 4 is selected from: -H, methyl, ethyl, isopropyl, tert-butyl, isobutyl One of radical and isopentyl, particularly preferably, said R 4 is -H.
  • said L is selected from: One of.
  • the D is a drug molecule
  • the drug molecule is selected from: amanitin, auristatin, calicheamicin, camptothecin, camptothecin derivatives and metabolites (SN-38 ), cryptomycin, daunomycin, dolastatin, doxorubicin, docarmycin, epothilone, espamycin, geldanamycin, maytansine, formazan Amhotrexate, monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), pyrrolobenzodiazepines, rhizobin, SG2285, tubulysin, vindesine, toxoids, or the above Derivatives of any kind;
  • the drug molecule is selected from one of: SN38 and its derivatives, Dxd, exitecan, MMAE, MMAF, MMAD, PBD and its derivatives, calicheamicin and TPL.
  • the drug molecule is selected from: One of them, preferably, the drug molecule is selected from:
  • the fourth aspect of the present invention is to provide an antibody-drug conjugate of general formula (IV).
  • Ab is an antibody, and said antibody includes monoclonal antibody and polyclonal antibody, preferably monoclonal antibody, more preferably internalized monoclonal antibody;
  • L1 is selected from: one of straight chain, Y-type and multi-branched polyethylene glycol residues, L1 includes but not limited to straight-chain double-ended PEG, Y-type PEG, 4-arm branched chain PEG, 6-arm branched Chain PEG or 8-arm branched chain PEG, preferably, L is a straight-chain polyethylene glycol residue, with a structure shown in general formula (III):
  • n is independently selected from an integer of 1-30, preferably an integer of 1-25, more preferably an integer of 4-24 (such as 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24).
  • said n1 is 8.
  • n 1 is 24.
  • L 1 is a Y-shaped and multi-branched polyethylene glycol residue, it can have one or more branches and connected.
  • L 2 is a linking group, for Wherein A is a peptide linker, and B is selected from: Wherein, Y 1 and Y 2 are each independently selected from: -H, halogen, -OC 1-10 alkyl, C 1-10 straight chain/branched chain alkyl, C 3-10 cycloalkyl, -OH, Preferably, both Y 1 and Y 2 are -H;
  • Y 3 and Y 4 are each independently selected from: -H, C 1-10 straight chain/branched chain alkyl, C 3-10 cycloalkyl, Preferably, both Y 3 and Y 4 are -CH 3 ;
  • B is
  • said B is
  • D is a drug molecule.
  • the peptide linker includes one or more identical or different polypeptide residues
  • the polypeptide residues include two or more identical or different amino acid residues or amino acid residue derivatives
  • Described amino acid is selected from: alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine acid, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, citrulline, ornithine, and cystine
  • the amino acid is selected from the group consisting of: valine, citrulline, glycine, alanine, phenylalanine, tyrosine, isoleucine, leucine acid and arginine or a combination of two or more
  • the polypeptide residues include valine-citrulline, valine-alanine and
  • R 1 , R 2 , R 3 , and R 4 are each independently selected from: -H, -C 1-6 straight chain/branched chain alkyl, -(C 1-6 alkylene )-OH, -(C 1-6 alkylene)-SH, -(CH 2 ) 1-6 -cycloalkyl, -(C 1-6 alkylene)-aryl ring, -(C 1- 6 alkylene)-heterocycloalkyl, -(C 1-6 alkylene)-heterocyclic aryl, -(C 1-6 alkylene)-COOH, -(C 1-6 alkylene) -CONH 2 , -(C 1-6 alkylene)-NH 2 and One of them, preferably, said R 1 , R 2 , R 3 , and R 4 are each independently selected from: -H, -C 1-6 straight chain/branched chain alkyl, -(C 1-6 alkylene )-OH, -(C 1-6 al
  • the R 1 is selected from one of: -H, methyl, ethyl, isopropyl, tert-butyl, isobutyl and isopentyl
  • the R 2 is selected from: -H , methyl, ethyl, isopropyl, tert-butyl, isobutyl, isopentyl and One of them
  • the R 3 is (C 1 alkylene)-phenyl, (C 2 alkylene)-phenyl, (C 3 alkylene)-phenyl, (C 4 alkylene )-phenyl, (C 5 alkylene)-phenyl and (C 6 alkylene)-phenyl
  • the R 4 is selected from: -H, methyl, ethyl, isopropyl
  • said R 1 is -H or isopropyl
  • said R 2 is selected from: -
  • said A is selected from:
  • the L2 is selected from: A combination of one or more of, said R 1 , R 2 , R 3 , R 4 are each independently selected from: -H, -C 1-6 straight chain/branched chain alkyl, -(C 1 -6 alkylene)-OH, -(C 1-6 alkylene)-SH, -(CH 2 ) 1-6 -cycloalkyl, -(C 1-6 alkylene)-aryl ring, -(C 1-6 alkylene)-heterocycloalkyl, -(C 1-6 alkylene)-heterocyclic aryl, -(C 1-6 alkylene)-COOH, -(C 1- 6 alkylene) -CONH 2 , -(C 1-6 alkylene) -NH 2 and One of them, preferably, the L2 is selected from: One of.
  • said R 1 is selected from: -H, -C 1-6 straight chain/branched chain alkyl, and one of (C 1-6 alkylene)-aromatic ring groups, more preferably, said R 1 is selected from: -H, methyl, ethyl, isopropyl, tert-butyl, isobutyl One of radical and isopentyl, particularly preferably, said R 1 is -H or isopropyl.
  • said R 2 is selected from: -H, -C 1-6 straight chain/branched chain alkyl, and (C 1-6 alkylene)-aryl ring group, more preferably, the R 2 is selected from: -H, methyl, ethyl, isopropyl, tert-butyl, isobutyl base, isopentyl and One of, particularly preferably, said R 2 is selected from: -H, methyl and One of.
  • said R 3 is (C 1-6 alkylene)-aromatic ring group, more preferably, said R 3 is (C 1 alkylene)-phenyl, (C 2 alkylene )-phenyl, (C 3 alkylene)-phenyl, (C 4 alkylene)-phenyl, (C 5 alkylene)-phenyl and (C 6 alkylene)-phenyl
  • said R 3 is (C 1 alkylene)-phenyl.
  • said R 4 is -H, -C 1-6 straight chain/branched chain alkyl, and one of (C 1-6 alkylene)-aromatic ring groups, more preferably, said R 4 is selected from: -H, methyl, ethyl, isopropyl, tert-butyl, isobutyl One of radical and isopentyl, particularly preferably, said R 4 is -H.
  • said L is selected from: One of.
  • the drug molecule is selected from: amanitin, auristatin, calicheamicin, camptothecin, camptothecin derivatives and metabolites (SN-38), cryptomycin, Dao Normycin, Dolastatin, Doxorubicin, Duocamycin, Epothilone, Espamycin, Geldanamycin, Maytansine, Methotrexate, Monomethylol Restatin E (MMAE), monomethyl auristatin F (MMAF), pyrrolobenzodiazepine, rhizobin, SG2285, tubulysin, vindesine, toxoid or any derivative of the above;
  • the drug molecule is selected from one of: SN38 and its derivatives, Dxd, exitecan, MMAE, MMAF, MMAD, PBD and its derivatives, calicheamicin and TPL.
  • the drug molecule is selected from: One of them, preferably, the drug molecule is selected from:
  • X is a linking group selected from -O-, -S-, C 1-12 straight chain/branched chain alkyl, C 3-12 cycloalkyl, C 6-12 aralkyl, and or a group consisting of combinations thereof, wherein R is selected from: -H, C 1-10 straight chain/branched alkyl;
  • L3 is selected from linear or branched C 1-12 alkylene, C 6-12 arylene, C 3-12 cycloalkylene, -S-, or a combination thereof;
  • L4 is selected from one of linear, Y-type and multi-branched polyethylene glycol residues
  • l is an integer selected from 1-50.
  • Q' is selected from L3 selected from L4 selected from n is selected from an integer of 1-30 (such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30), preferably, n is an integer selected from 2-15.
  • n 4.
  • n 12.
  • the antibody drug conjugate has a structure shown in formula (V):
  • Ab is an antibody
  • said antibody includes monoclonal antibody and polyclonal antibody, preferably monoclonal antibody, more preferably internalized monoclonal antibody.
  • the form of the antibody can be, for example: chimeric antibody, humanized antibody, human antibody, antibody fragment (Fab, Fab', F(ab)2, F(ab)2) that can bind to antigen, subunit Fragments (single-chain constructs) or antibody Fc fusion proteins, etc., preferably, the monoclonal antibody is reactive to antigens or epitopes associated with cancer, malignant cells, infectious organisms or autoimmune diseases.
  • the monoclonal antibody is selected from: anti-HER2 antibody, anti-EGFR antibody, anti-PMSA antibody, anti-VEGFR antibody, anti-CD20 antibody, anti-CD30 antibody, anti-FR ⁇ antibody, anti-CD22 antibody , anti-CD56 antibody, anti-CD29 antibody, anti-GPNMB antibody, anti-CD138 antibody, anti-CD74 antibody, anti-ENPP3 antibody, anti-Nectin-4 antibody, anti-EGFRVIII antibody, anti-SLC44A4 antibody, anti-mesothelin antibody (anti-mesothelin antibody), Anti-ET8R antibody, anti-CD37 antibody, anti-CEACAM5 antibody, anti-CD70 antibody, anti-MUC16 antibody, anti-CD79b antibody, anti-MUC16 antibody, anti-Muc1 antibody.
  • the antigen is selected from: HER-2/neu, EGFR, FR ⁇ , Nectin-4, carbonic anhydrase IX, B7, CCCL19, CCCL21, CSAp, BrE3, CD1, CD1a, CD2, CD3, CD4, CD5, CD8, CD11A, CD14, CD15, CD16, CD18, CD19, CD20, CD21, CD22, CD23, CD25, CD29, CD30, CD32b, CD33, CD37, CD38, CD40, CD40L, CD44, CD45, CD46, CD52, CD54, CD55, CD59, CD64, CD67, CD70, CD74, CD79a, CD80, CD83, CD95, CD126, CD133, CD138, CD147, CD154, CEACAM5, CEACAM-6, alpha-fetoprotein (AFP) , VEGF, ED-B fibronectin, EGP-1, EGP-2, EGF receptor (Erb
  • hypoxia-inducible factor HIF
  • HM1.24 insulin-like growth factor (ILGF)
  • IFN- ⁇ IFN- ⁇
  • IFN- ⁇ IFN- ⁇
  • IFN- ⁇ IFN- ⁇
  • IL-2R IL-4R
  • IL-6R IL-13R
  • IL-15R IL-17R
  • IL-18R IL-2, IL-6, IL-8, IL-12, IL-15, IL-17, IL-18, IL-25
  • IP-10 IGF- 1R, Ia, HM1.24, ganglioside, HCG, HLA-DR, CD66a-d, MAGE, mCRP, MCP-1, MIP-1A, MIP-1B, macrophage migration inhibitory factor (MIF), MUC1, MUC2, MUC3, MUC4, MUC5, placental growth factor (PIGF), PSA, PSMA, PSMA dimer, PAM4 antigen, NCA-95, NCA-90, A3, A33, Ep-CAM, KS-1
  • the fifth aspect of the present invention is to provide a preparation method of the antibody-drug conjugate of general formula (V).
  • Ab is an antibody, and said antibody includes monoclonal antibody and polyclonal antibody, preferably monoclonal antibody, more preferably internalized monoclonal antibody;
  • L1 is selected from: one of straight chain, Y-type and multi-branched polyethylene glycol residues, L1 includes but not limited to straight-chain double-ended PEG, Y-type PEG, 4-arm branched chain PEG, 6-arm branched Chain PEG or 8-arm branched chain PEG, when L 1 is a Y-type and multi-branched polyethylene glycol residue, it can have one or more branches and connected.
  • L is a straight-chain polyethylene glycol residue with a structure shown in general formula (III):
  • n is independently selected from an integer of 1-30, preferably an integer of 1-25, more preferably an integer of 4-24 (such as 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24).
  • said n1 is 8.
  • n 1 is 24.
  • L 2 is a linking group, for Wherein A is a peptide linker, and B is selected from: Wherein, Y 1 and Y 2 are each independently selected from: -H, halogen, -OC 1-10 alkyl, C 1-10 straight chain/branched chain alkyl, C 3-10 cycloalkyl, -OH, Preferably, both Y 1 and Y 2 are -H;
  • Y 3 and Y 4 are each independently selected from: -H, C 1-10 straight chain/branched chain alkyl, C 3-10 cycloalkyl, Preferably, both Y 3 and Y 4 are -CH 3 ;
  • B is
  • said B is
  • D is a drug molecule.
  • the peptide linker includes one or more identical or different polypeptide residues
  • the polypeptide residues include two or more identical or different amino acid residues or amino acid residue derivatives
  • Described amino acid is selected from: alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine acid, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, citrulline, ornithine, and cystine
  • the amino acid is selected from the group consisting of: valine, citrulline, glycine, alanine, phenylalanine, tyrosine, isoleucine, leucine acid and arginine or a combination of two or more
  • the polypeptide residues include valine-citrulline, valine-alanine and
  • R 1 , R 2 , R 3 , and R 4 are each independently selected from: -H, -C 1-6 straight chain/branched chain alkyl, -(C 1-6 alkylene )-OH, -(C 1-6 alkylene)-SH, -(CH 2 ) 1-6 -cycloalkyl, -(C 1-6 alkylene)-aryl ring, -(C 1- 6 alkylene)-heterocycloalkyl, -(C 1-6 alkylene)-heterocyclic aryl, -(C 1-6 alkylene)-COOH, -(C 1-6 alkylene) -CONH 2 , -(C 1-6 alkylene)-NH 2 and One of them, preferably, said R 1 , R 2 , R 3 , and R 4 are each independently selected from: -H, -C 1-6 straight chain/branched chain alkyl, -(C 1-6 alkylene )-OH, -(C 1-6 al
  • the R 1 is selected from one of: -H, methyl, ethyl, isopropyl, tert-butyl, isobutyl and isopentyl
  • the R 2 is selected from: -H , methyl, ethyl, isopropyl, tert-butyl, isobutyl, isopentyl and One of them
  • the R 3 is (C 1 alkylene)-phenyl, (C 2 alkylene)-phenyl, (C 3 alkylene)-phenyl, (C 4 alkylene )-phenyl, (C 5 alkylene)-phenyl and (C 6 alkylene)-phenyl
  • the R 4 is selected from: -H, methyl, ethyl, isopropyl
  • said R 1 is -H or isopropyl
  • said R 2 is selected from: -
  • said A is selected from:
  • the L2 is selected from: A combination of one or more of, said R 1 , R 2 , R 3 , R 4 are each independently selected from: -H, -C 1-6 straight chain/branched chain alkyl, -(C 1 -6 alkylene)-OH, -(C 1-6 alkylene)-SH, -(CH 2 ) 1-6 -cycloalkyl, -(C 1-6 alkylene)-aryl ring, -(C 1-6 alkylene)-heterocycloalkyl, -(C 1-6 alkylene)-heterocyclic aryl, -(C 1-6 alkylene)-COOH, -(C 1- 6 alkylene) -CONH 2 , -(C 1-6 alkylene) -NH 2 and One of them, preferably, the L2 is selected from: One of.
  • said R 1 is selected from: -H, -C 1-6 straight chain/branched chain alkyl, and one of (C 1-6 alkylene)-aromatic ring groups, more preferably, said R 1 is selected from: -H, methyl, ethyl, isopropyl, tert-butyl, isobutyl One of radical and isopentyl, particularly preferably, said R 1 is -H or isopropyl.
  • said R 2 is selected from: -H, -C 1-6 straight chain/branched chain alkyl, and (C 1-6 alkylene)-aryl ring group, more preferably, the R 2 is selected from: -H, methyl, ethyl, isopropyl, tert-butyl, isobutyl base, isopentyl and One of, particularly preferably, said R 2 is selected from: -H, methyl and One of.
  • said R 3 is (C 1-6 alkylene)-aromatic ring group, more preferably, said R 3 is (C 1 alkylene)-phenyl, (C 2 alkylene )-phenyl, (C 3 alkylene)-phenyl, (C 4 alkylene)-phenyl, (C 5 alkylene)-phenyl and (C 6 alkylene)-phenyl
  • said R 3 is (C 1 alkylene)-phenyl.
  • said R 4 is -H, -C 1-6 straight chain/branched chain alkyl, and one of (C 1-6 alkylene)-aromatic ring groups, more preferably, said R 4 is selected from: -H, methyl, ethyl, isopropyl, tert-butyl, isobutyl One of radical and isopentyl, particularly preferably, said R 4 is -H.
  • said L is selected from: One of.
  • the D is a drug molecule
  • the drug molecule is selected from: amanitin, auristatin, calicheamicin, camptothecin, camptothecin derivatives and metabolites (SN-38 ), cryptomycin, daunomycin, dolastatin, doxorubicin, docarmycin, epothilone, espamycin, geldanamycin, maytansine, formazan Amhotrexate, monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), pyrrolobenzodiazepines, rhizobin, SG2285, tubulysin, vindesine, toxoids, or the above Derivatives of any kind;
  • the drug molecule is selected from one of: SN38 and its derivatives, Dxd, exitecan, MMAE, MMAF, MMAD, PBD and its derivatives, calicheamicin and TPL.
  • the drug molecule is selected from: One of them, preferably, the drug molecule is selected from:
  • the sixth aspect of the present invention is to provide an antibody drug conjugate of general formula (IV) or conjugate 1 of general formula (II) or conjugate 2 of general formula (VI) in disease prevention and/or or therapeutic drug applications.
  • the disease is cancer, pathogenic organism infection or autoimmune disease.
  • the cancer is hematopoietic tumor, carcinoma, sarcoma, melanoma or glial tumor.
  • the pathogenic organism is selected from the group consisting of: human immunodeficiency virus (HIV), Mycobacterium tuberculosis, Streptococcus agalactiae, methicillin-resistant Staphylococcus aureus, Legionella pneumophila, pyogenes Streptococcus, Escherichia coli, Neisseria gonorrhoeae, Neisseria meningitidis, Pneumococcus spp, Haemophilus influenzae type B, Treponema pallidum, Lyme disease, West Nile virus, Pseudomonas aeruginosa , Mycobacterium leprae, Bacillus abortus, rabies virus, influenza virus, cytomegalovirus, type 1 herpes simplex virus, type 2 herpes simplex virus, human serum parvovirus, respiratory syncytial virus, varicella-zoster virus, B Hepatitis virus, mea
  • the autoimmune disease is selected from the group consisting of immune-mediated thrombocytopenia, dermatomyositis, Sjögren's syndrome, multiple sclerosis, Sidenham's chorea, myasthenia gravis , systemic lupus erythematosus, lupus nephritis, rheumatic fever, rheumatoid arthritis, polyglandular syndrome, bullous pemphigoid, diabetes mellitus, Henschler's purpura, poststreptococcal nephritis, nodules Erythema dermatitis, Takayasu arteritis, Addison's disease, sarcoidosis, ulcerative colitis, erythema multiforme, IgA nephropathy, polyarteritis nodosa, ankylosing spondylitis, Goodpasture syndrome , thromboangiitis obliterans, primary bil
  • the seventh aspect of the present invention provides a method for disease treatment, the method comprising administering the antibody drug conjugate of the present invention to a subject.
  • the antibody-drug conjugate is administered in combination with one or more treatment methods selected from the following: unconjugated antibody, radiolabeled antibody, drug-conjugated antibody, toxin-conjugated antibody, gene therapy, chemotherapy, Therapeutic peptides, oligonucleotides, local radiation, surgery and interfering RNA therapy.
  • the disease is cancer, pathogenic organism infection or autoimmune disease.
  • the cancer is hematopoietic tumor, carcinoma, sarcoma, melanoma or glial tumor.
  • the pathogenic organism is selected from the group consisting of: human immunodeficiency virus (HIV), Mycobacterium tuberculosis, Streptococcus agalactiae, methicillin-resistant Staphylococcus aureus, Legionella pneumophila, pyogenes Streptococcus, Escherichia coli, Neisseria gonorrhoeae, Neisseria meningitidis, Pneumococcus spp, Haemophilus influenzae type B, Treponema pallidum, Lyme disease, West Nile virus, Pseudomonas aeruginosa , Mycobacterium leprae, Bacillus abortus, rabies virus, influenza virus, cytomegalovirus, type 1 herpes simplex virus, type 2 herpes simplex virus, human serum parvovirus, respiratory syncytial virus, varicella-zoster virus, B Hepatitis virus, mea
  • the autoimmune disease is selected from the group consisting of immune-mediated thrombocytopenia, dermatomyositis, Sjögren's syndrome, multiple sclerosis, Sidenham's chorea, myasthenia gravis , systemic lupus erythematosus, lupus nephritis, rheumatic fever, rheumatoid arthritis, polyglandular syndrome, bullous pemphigoid, diabetes mellitus, Henschler's purpura, poststreptococcal nephritis, nodules Erythema dermatitis, Takayasu arteritis, Addison's disease, sarcoidosis, ulcerative colitis, erythema multiforme, IgA nephropathy, polyarteritis nodosa, ankylosing spondylitis, Goodpasture syndrome , thromboangiitis obliterans, primary bil
  • the novel linker-drug conjugates provided by the present invention can be coupled with antibodies through simple chemical methods. Compared with traditional antibody-drug conjugates, the DAR value distribution of the conjugates obtained by using this linker is very narrow. Therefore, the uniformity of the product generated is high, and the obtained cross-linked product has a single distribution component (DAR is 4) accounting for more than 80%; meanwhile, the antibody-drug conjugate provided by the present invention, naked antibody and ADC with low cross-linking degree The proportion is almost zero (mass spectrometry can not detect components with DAR of 0 and 1); this antibody drug conjugate can not only target the EGFR antigen, but also has a strong activity of killing tumor cells.
  • the antibody-drug conjugate of the present invention has certain safety and effectiveness in treating tumors, and the hydrophilicity endowed by ethylene glycol after coupling can be used to adjust the characteristics of biomolecules ;
  • the in vitro tumor cell proliferation inhibitory activity of the conjugate is improved or maintained compared with the traditional mc-VC-PAB coupling biological activity, drug metabolism stability, safety and other pharmaceutical properties;
  • the PEG chain is introduced into the method, which increases the solubility of cytotoxic drugs, makes the ADC coupling reaction more complete, and the coupling efficiency is higher; the coupling method provided by the invention is suitable for most antibodies, so it has a wide application prospect; Compared with the
  • C 1-10 straight chain/branched chain alkyl described in the present invention includes methyl, ethyl, C 3 straight chain/branched chain alkyl, C 4 straight chain/branched chain alkyl, C 5 straight chain /branched chain alkyl, C 6 straight chain/branched chain alkyl, C 7 straight chain/branched chain alkyl, C 8 straight chain/branched chain alkyl, C 9 straight chain/branched chain alkyl, C 10 straight chain / branched chain alkyl.
  • C 1-12 straight chain/branched chain alkyl described in the present invention includes methyl, ethyl, C 3 straight chain/branched chain alkyl, C 4 straight chain/branched chain alkyl, C 5 straight chain /branched chain alkyl, C 6 straight chain/branched chain alkyl, C 7 straight chain/branched chain alkyl, C 8 straight chain/branched chain alkyl, C 9 straight chain/branched chain alkyl, C 10 straight chain /branched chain alkyl, C 11 straight chain/branched chain alkyl, C 12 straight chain/branched chain alkyl.
  • C 1-5 straight chain/branched chain alkyl described in the present invention includes methyl, ethyl, C 3 straight chain/branched chain alkyl, C 4 straight chain/branched chain alkyl, C 5 straight chain / branched chain alkyl.
  • C 3-12 cycloalkyl described in the present invention includes C 3 cycloalkyl, C 4 cycloalkyl, C 5 cycloalkyl, C 6 cycloalkyl, C 7 cycloalkyl, C 8 cycloalkane Group, C 9 cycloalkyl, C 10 cycloalkyl, C 11 cycloalkyl, C 12 cycloalkyl.
  • C6-12 aromatic ring group described in the present invention includes C6 aromatic ring group (phenyl), C7 aromatic ring group, C8 aromatic ring group, C9 aromatic ring group, C10 aromatic ring group, C 11 aromatic ring group, C 12 aromatic ring group.
  • SN38 described in the present invention refers to 7-ethyl-10-hydroxycamptothecin.
  • MMAE described in the present invention refers to monomethyl auristatin E.
  • MMAF described in the present invention refers to monomethylauristatin F.
  • MMAD described in the present invention refers to monomethyl auristatin D.
  • PBD described in the present invention refers to pyrrolobenzodiazepines.
  • TPL mentioned in the present invention refers to triptolide.
  • Dxd described in the present invention refers to derutecan.
  • Figure 1 is the mass spectrum of Compound 1.
  • Figure 2 is the NMR image of compound 1.
  • Figure 3 is the mass spectrum of compound 6.
  • Figure 4 is the NMR image of compound 6.
  • Figure 5 is the mass spectrum of compound 7.
  • Figure 6 is the NMR image of compound 7.
  • Figure 7 is the mass spectrum of compound 8.
  • Figure 8 is the NMR image of compound 8.
  • Figure 9 is the mass spectrum of compound 9.
  • Figure 10 is the NMR image of compound 9.
  • Figure 11 is the mass spectrum of compound 10.
  • Figure 12 is the NMR image of compound 10.
  • Figure 13 is the mass spectrum of compound 11.
  • Figure 14 is the NMR image of compound 11.
  • the raw material Fmoc-VC-PAB (9.0g, 15.0mmol) was dissolved in 30mL dimethyl sulfoxide, diethylamine (2.2g, 30.0mmol) was added, and the reaction was stirred at room temperature for 2h, and the reaction was monitored by the liquid phase. , 200mL of dichloromethane was added, a large amount of solids appeared, stirred for 30min, and the solids were collected by filtration to obtain product 1f (5.0g), with a yield of 88%.
  • the raw material Fmoc-VA-PAB (7.7g, 15.0mmol) was dissolved in 30mL of dimethyl sulfoxide, diethylamine (2.2g, 30.0mmol) was added, and the reaction was stirred at room temperature for 2h, and the reaction was monitored by the liquid phase. , 200mL of dichloromethane was added, a large amount of solids appeared, stirred for 30min, and the solids were collected by filtration to obtain product 2f (3.9g), with a yield of 90%.
  • Boc-SN38 (984 mg, 2 mmol) and 4-dimethylaminopyridine (366 mg, 3 mmol) in 8 mL of dichloromethane, add triphosgene (268 mg) under nitrogen protection, stir at room temperature for 5 minutes, and add A dichloromethane solution (2 mL) of Boc-DMEA (456 mg, 2.5 mmol) was stirred at room temperature for 5 minutes, and LCMS showed that the starting material was completely reacted. The reaction solution was washed twice with water to obtain the crude product 5a (yellow solid, 543mg), with a yield of 77%.
  • the raw material Fmoc-VC-PAB (9.0g, 15.0mmol) was dissolved in 30mL dimethyl sulfoxide, diethylamine (2.2g, 30.0mmol) was added, and the reaction was stirred at room temperature for 2h, and the reaction was monitored by the liquid phase. , 200mL of dichloromethane was added, a large amount of solids appeared, stirred for 30min, and the solids were collected by filtration to obtain product 7c (5.0g), with a yield of 88%.
  • Antibody reacts with PEG derivatives containing N3 functional group and NHS functional group, so that the antibody is labeled with N3 functional group.
  • the reaction was carried out according to the molar ratio of mAb to compound 6 which was 1:20.
  • First measure 208 ⁇ L of mAb solution (concentration: 9.6 mg/mL) and place it in a 1.5 mL centrifuge tube, then the measured mAb is 2 mg.
  • the reaction was carried out according to the molar ratio of mAb to compound 10 of 1:20. First measure 1 mL of mAb solution (concentration: 2.0 mg/mL) and place it in a 1.5 mL centrifuge tube, then the measured mAb is 2 mg. A 5.21 mg/mL compound 11 solution was prepared in DMSO, and 100 ⁇ L was added to a centrifuge tube, mixed well, and oscillated for 2 h. After the reaction was completed, unreacted compound 11 was removed by ultrafiltration to obtain ADC4.
  • the DAR value of the ADC prepared in Example 12 was detected by a liquid phase method.
  • Example 12 Dissolve the ADC1, ADC2, ADC3, and ADC4 prepared in Example 12, and detect ADC1, ADC2, ADC3, and ADC4 according to the liquid chromatography conditions in Table 2. Under the liquid chromatography conditions, SN38 is separated from ADC1, ADC2, ADC3, and ADC4 degree greater than 1.5. Prepare standard curves of different concentrations, establish a linear standard curve between the peak area of SN38 and the concentration, and quantify SN38 on ADC 1, ADC 2, ADC3 and ADC4, respectively. According to the antibody concentration of ADC and the concentration of SN38, the conjugation number of SN38 on ADC was calculated.
  • the DAR value of ADC 1 prepared in Example 12 is 4, the DAR value of ADC2 is 4, the DAR value of ADC3 is 2, the DAR value of ADC4 is 4, and the DAR values of ADC1, ADC2 and ADC4 are all higher than ADC3.
  • the results show that compared with the linker without PEG group, the DAR values of ADC1, ADC2 and ADC4 obtained by the invention are higher.
  • the affinity of the ADC drug is detected by the method of SPR, and the test instrument is biacore, a product of GE Company.
  • the simple operation steps are: coupling the EGFR antigen to the CM chip.
  • the results showed that ADC1, ADC2 and ADC3 prepared in Example 12 had less obvious decrease in the affinity of the antibodies, and the affinity of ADC1 and ADC2 was higher than that of ADC3.
  • Cell survival rate subtract the background OD value (blank group) from the OD value of each test well, and take the mean ⁇ SD of the OD value of each repeated well.
  • Cell viability% (OD of drug-added cells/OD of control cells) ⁇ 100%.
  • the IC50 value of ADC1 and ADC2 prepared by embodiment 12 is all less than the IC50 value of ADC3, less than the IC50 value of SN38.
  • the ADC1 and ADC2 containing PEG linker obtained by the present invention have better activity.

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Abstract

Conjugué et conjugué anticorps-médicament préparé à l'aide de celui-ci. Une conjugaison dirigée vers un site d'un nouveau conjugué lieur-médicament fourni pour un groupe azoture modifié sur un anticorps peut être obtenue au moyen d'une simple réaction SPAAC, donnant un conjugué anticorps-médicament hautement homogène ; de plus, le conjugué anticorps-médicament fourni présente une grande homogénéité et peut non seulement cibler un antigène EGFR, mais présente également une forte activité de destruction des cellules tumorales. Par rapport au necitumumab lui-même, le conjugué anticorps-médicament n'affecte pas l'affinité, l'activité d'endocytose et la propriété de ciblage d'un anticorps, et conserve bien sa fonction biologique. Dans le cadre d'une évaluation de son activité in vitro, par rapport au SN38, le conjugué anticorps-médicament présente une activité d'inhibition tumorale significativement améliorée, au niveau nM de la CI50.
PCT/CN2022/129983 2021-12-30 2022-11-04 Conjugué et conjugué anticorps-médicament préparé à l'aide de celui-ci WO2023124537A1 (fr)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102448494A (zh) * 2009-02-13 2012-05-09 免疫医疗公司 具有胞内可裂解的键的免疫共轭物
WO2020229982A1 (fr) * 2019-05-10 2020-11-19 Takeda Pharmaceutical Company Limited Conjugués anticorps-médicament
CN112533951A (zh) * 2018-05-09 2021-03-19 里珍纳龙药品有限公司 抗msr1抗体及其使用方法
CN113382985A (zh) * 2018-11-20 2021-09-10 里珍纳龙药品有限公司 用作lxr激动剂的双八氢菲羧酰胺类衍生物及其蛋白质偶联物
WO2021248048A2 (fr) * 2020-06-05 2021-12-09 Development Center For Biotechnology Conjugués anticorps-médicament contenant un anticorps anti-mésothéline et leurs utilisations

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102448494A (zh) * 2009-02-13 2012-05-09 免疫医疗公司 具有胞内可裂解的键的免疫共轭物
CN112533951A (zh) * 2018-05-09 2021-03-19 里珍纳龙药品有限公司 抗msr1抗体及其使用方法
CN113382985A (zh) * 2018-11-20 2021-09-10 里珍纳龙药品有限公司 用作lxr激动剂的双八氢菲羧酰胺类衍生物及其蛋白质偶联物
WO2020229982A1 (fr) * 2019-05-10 2020-11-19 Takeda Pharmaceutical Company Limited Conjugués anticorps-médicament
WO2021248048A2 (fr) * 2020-06-05 2021-12-09 Development Center For Biotechnology Conjugués anticorps-médicament contenant un anticorps anti-mésothéline et leurs utilisations

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