Linker based on benzoquinone structure, antibody coupling medicine and application of linker and antibody coupling medicine

Abstract

The invention relates to a linker containing a benzoquinone structure, an antibody coupling medicine containing the linker and application of the linker, further relates to a pharmaceutical composition containing the antibody coupling medicine, and further relates to application of the antibody coupling medicine for treating and / or preventing diseases.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of medicinal chemistry, and specifically relates to a linker comprising a benzoquinone structure, an antibody conjugate drug comprising the linker, and the use of the linker, and further relates to a pharmaceutical composition comprising the antibody conjugate drug, and further relates to the use of the antibody conjugate drug for treating and / or preventing diseases. BACKGROUND

[0002] An antibody-drug conjugate (ADC) is a new type of drug form, which is composed of three parts, namely an antibody, a linker, and a small molecule drug, wherein the antibody functions to achieve targeting, the small molecule drug functions to exert a treatment on a disease, and the linker functions to realize the organic combination of the antibody and the cytotoxic structure, so as to form an organic whole. The antibody-drug conjugate can combine the targeting of the antibody and the high activity of the small molecule drug, and has the advantages of strong targeting, high activity, small toxic and side effects, and long half-life.

[0003] Among the three components of the ADC drug, the structure of the linker has a very important influence on the properties of the entire ADC drug. The properties of the linker determine the metabolic properties, effectiveness and safety of the ADC drug. An ideal linker should have the properties of remaining stable in the blood circulation system and rapidly and effectively releasing the small molecule drug after reaching the target tissue. The research and development of the linker chain becomes an important part of the research and development of the ADC drug.

[0004] At present, the linker of the ADC drug can be divided into cleavable and non-cleavable types. Among the ADC drugs that have been marketed, 80% of the ADC drugs adopt the cleavable linker chain. The current cleavable linker chain mainly utilizes the difference between the tumor microenvironment (pH value, overexpressed substances, overexpressed enzymes) and normal sites to achieve specific cleavage of the ADC linker chain. According to the different drug release principles, the ADC linker can be divided into pH-responsive (hydrazone bond), oxidation-reduction type (overexpressed glutathione-sensitive disulfide bond), enzyme-responsive type (tissue protease-sensitive peptide chain, phospholipase-sensitive phospholipid bond, galactosidase-sensitive glycoside bond).

[0005] Among the ADC drugs that have been approved for marketing and are in different research stages, the linker based on tissue protease is used the most, but the existing research results show that tissue protease is expressed in normal tissues and tumor tissues, and it widely exists in the lysosomes of most cells of mammals, and the expression level of tissue protease in normal tissues and tumor tissues is not significantly different. For the inevitable off-target ADC, the tissue protease existing in the normal tissues will also promote the release of the cytotoxic drug in the ADC, leading to the toxic and side effects of the ADC on the normal tissues. New ADC drugs need to be developed. SUMMARY

[0006] Based on the significant difference in microenvironment between tumor tissue and normal tissue, a new linker is designed using enzymes with more obvious expression level difference, which is expected to overcome the above defects and provide a powerful supplement for ADC research. Quinone oxidoreductase (DT-diaphorase) can use NADH (coenzyme) or reduced coenzyme II (NADPH) as an electron donor to catalyze the conversion of quinone compounds into hydroquinone. Compared with normal tissue cells, the level of quinone oxidoreductase in tumor tissue sites is significantly improved. Its content in tumor tissue is usually several times or even dozens of times that of normal tissue, for example, its content in lung tumor sites, liver tumor sites, colon cancer sites and breast tumor sites is 12 to 18 times, 4 to 19 times, 3 to 4 times and 3 times more than that of normal tissue, respectively.

[0007] The inventors of the present application prepared a linker containing a benzoquinone structure, which can be specifically cleaved in the presence of quinone oxidoreductase 1 (NQO1), and also prepared an antibody conjugate drug containing the linker, which can be specifically cleaved to release the active drug under the condition of quinone oxidoreductase 1 (NQO1) and can remain stable without NQO1.

[0008] Therefore, the first aspect of the present application provides a compound containing a benzoquinone structure represented by Formula I or a racemate, tautomer, stereoisomer, enantiomer, diastereomer, or mixture of the foregoing configurations thereof, or a pharmaceutically acceptable salt or solvate or hydrate thereof, wherein

[0009]

[0010] R1 is hydrogen or C 1-6 alkyl;

[0011] R2 is hydrogen or C 1-6 alkyl;

[0012] X1 is -O-, -S- or NR7-, wherein R7 is hydrogen or C 1-6 alkyl;

[0013] Z is -CH2-, -CH(CH3)-, -C(CH3)2- or -CH2-NH-;

[0014] R3 is hydroxyl, halogen or wherein R6 is hydrogen, halogen, C 1-6 alkyl, nitro or C 1-6 alkoxy, p is 0, 1, 2, 3 or 4;

[0015] R5 is hydrogen, halogen, C 1-6 alkyl, nitro or C 1-6alkoxy; r is 0, 1, 2, 3, or 4;

[0016] X2is -0-, -S-, or NR8-, wherein R8is hydrogen or C 1-6 alkyl;

[0017] L is -(CH2) m -C(O)NH-(CH2CH2O) n -(CH2)2-C(O)NH-(CH2) t -C(O)-, -(CH2) m -C(O)NH-(CH2CH2O) n -C(O)NH-(CH2) t -C(O)-, -(CH2CH2O) n -(CH2) m -C(O)-, -(CH2) m -(CH2CH2O) n -C(O)-, or -(CH2) m -C(O)NH-(CH2) t -C(O)-, wherein each t is independently 1, 2, 3, or 4, each m is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12, each n is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; and

[0018] R4is

[0019] The second aspect of the present application provides use of the compound of the first aspect of the present application or a racemate, a tautomer, a stereoisomer, an enantiomer, a diastereomer, or a mixture of the foregoing forms of the present application, or a pharmaceutically acceptable salt or solvate or hydrate thereof, in the manufacture of an antibody conjugated drug.

[0020] The third aspect of the present application provides a conjugate or a racemate, a tautomer, a stereoisomer, an enantiomer, a diastereomer, or a mixture of the foregoing forms of the present application, or a pharmaceutically acceptable salt or solvate or hydrate thereof, which is obtained by conjugation reaction of the compound of the first aspect of the present application or a racemate, a tautomer, a stereoisomer, an enantiomer, a diastereomer, or a mixture of the foregoing forms of the present application, or a pharmaceutically acceptable salt or solvate or hydrate thereof with an active compound.

[0021] The fourth aspect of the present application provides use of the conjugate according to the third aspect of the present application or a racemate, a tautomer, a stereoisomer, an enantiomer, a diastereomer, or a mixture of the aforementioned forms of the conjugate, or a pharmaceutically acceptable salt or a solvate or a hydrate thereof in the preparation of an antibody conjugate drug.

[0022] The fifth aspect of the present application provides an antibody conjugate drug or a racemate, a tautomer, a stereoisomer, an enantiomer, a diastereomer, or a mixture of the aforementioned forms of the antibody conjugate drug, or a pharmaceutically acceptable salt or a solvate or a hydrate thereof, which is obtained by conjugation reaction of the conjugate according to the third aspect of the present application or a racemate, a tautomer, a stereoisomer, an enantiomer, a diastereomer, or a mixture of the aforementioned forms of the conjugate, or a pharmaceutically acceptable salt or a solvate or a hydrate thereof with an antibody or an antigen-binding fragment thereof.

[0023] The sixth aspect of the present application provides a pharmaceutical composition comprising at least one antibody conjugate drug according to the fifth aspect of the present application or a racemate, a tautomer, a stereoisomer, an enantiomer, a diastereomer, or a mixture of the aforementioned forms of the antibody conjugate drug, or a pharmaceutically acceptable salt or a solvate or a hydrate thereof, and one or more pharmaceutically acceptable carriers or excipients.

[0024] The seventh aspect of the present application provides use of the antibody conjugate drug according to the fifth aspect of the present application or a racemate, a tautomer, a stereoisomer, an enantiomer, a diastereomer, or a mixture of the aforementioned forms of the antibody conjugate drug, or a pharmaceutically acceptable salt or a solvate or a hydrate thereof in the preparation of a medicament, wherein the medicament is used for treating or alleviating the severity of a disease or a disorder in a subject, the disease or disorder being selected from the group consisting of a tumor, a bacterial infection, an infectious disease, a hematological disease, a metabolic disease, and an inflammation. DETAILED DESCRIPTION

[0026] Linkers comprising a benzoquinone structure

[0027] The first aspect of the present application provides a compound comprising a quinone structure represented by Formula I or a racemate, a tautomer, a stereoisomer, an enantiomer, a diastereomer, or a mixture of the aforementioned forms of the compound, or a pharmaceutically acceptable salt or a solvate or a hydrate thereof, wherein

[0028]

[0029] R1is hydrogen or C 1-6 alkyl;

[0030] R2is hydrogen or C 1-6 alkyl;

[0031] X1is -O-, -S-, or NR7-, wherein R7is hydrogen or C 1-6 alkyl;

[0032] Z is -CH2-, -CH(CH3)-, -C(CH3)2-, or -CH2-NH-;

[0033] R3is hydroxyl, halogen, or wherein R6is hydrogen, halogen, C 1-6 alkyl, nitro, or C 1-6 alkoxy, p is 0, 1, 2, 3, or 4;

[0034] R5is hydrogen, halogen, C 1-6 alkyl, nitro, or C 1-6 alkoxy; r is 0, 1, 2, 3, or 4;

[0035] X2is -O-, -S-, or NR8-, wherein R8is hydrogen or C 1-6 alkyl;

[0036] L is -(CH2) m -C(O)NH-(CH2CH2O) n -(CH2)2-C(O)NH-(CH2) t -C(O)-, -(CH2) m -C(O)NH-(CH2CH2O) n -C(O)NH-(CH2) t -C(O)-, -(CH2CH2O) n -(CH2) m -C(O)-, -(CH2) m -(CH2CH2O) n -C(O)-, or -(CH2) m -C(O)NH-(CH2) t -C(O)-, wherein each t is independently 1, 2, 3, or 4, each m is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12, each n is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; and

[0037] R4is

[0038] In certain embodiments, R1is hydrogen or C 1-4 alkyl.

[0039] In certain embodiments, R2is hydrogen or C 1-4 alkyl.

[0040] In certain embodiments, X1is -O- or -NR7-, wherein R7is as defined in any embodiment herein.

[0041] In certain embodiments, Z is -CH2- or -CH2-NH-.

[0042] In certain embodiments, R3is hydroxy, fluoro, chloro, bromo, iodo, or wherein R6and p are as previously defined.

[0043] In certain embodiments, R5is hydrogen, fluoro, chloro, bromo, iodo, C 1-4 alkyl, nitro, or C 1-4 alkoxy.

[0044] In certain embodiments, X2is -O- or -NR8-, wherein R8is as defined in any embodiment herein.

[0045] In certain embodiments, L is -(CH2) m -C(O)NH-(CH2CH2O) n -(CH2)2-C(O)NH-(CH2) t -C(O)-, -(CH2) m -C(O)NH-(CH2CH2O) n -C(O)NH-(CH2) t -C(O)-, -(CH2CH2O) n -(CH2) m -C(O)-, -(CH2) m -(CH2CH2O) n -C(O)- or -(CH2) m -C(O)NH-(CH2) t -C(O)-, wherein t, m, n are as previously defined.

[0046] In certain embodiments, R4is

[0047] In certain embodiments, R1is hydrogen, methyl, ethyl, n-propyl, i-propyl, n-butyl, sec-butyl, i-butyl, t-butyl. In certain embodiments, R1is hydrogen, methyl, ethyl, or n-propyl. In certain embodiments, R1is hydrogen, methyl, or ethyl. In certain embodiments, R1is methyl.

[0048] In certain embodiments, R2is hydrogen, methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl, t-butyl. In certain embodiments, R2is hydrogen, methyl, ethyl, or n-propyl. In certain embodiments, R2is hydrogen, methyl, or ethyl. In certain embodiments, R2is methyl.

[0049] In certain embodiments, X1is -NR7-, wherein R7is as defined in any embodiment herein.

[0050] In certain embodiments, Z is -CH2-.

[0051] In certain embodiments, R3is chloro or wherein R6and p are as defined in any embodiment herein.

[0052] In certain embodiments, R5is hydrogen, fluoro, chloro, bromo, iodo, methoxy, ethoxy, propoxy, butoxy, methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl, or t-butyl. In certain embodiments, R5is hydrogen, fluoro, chloro, bromo, iodo, methoxy, ethoxy, propoxy, methyl, ethyl, n-propyl, or i-propyl. In certain embodiments, R5is hydrogen, fluoro, chloro, bromo, methoxy, ethoxy, methyl, or ethyl. In certain embodiments, R5is hydrogen.

[0053] In certain embodiments, X2is -NR8-, wherein R8is as defined in any embodiment herein.

[0054] In certain embodiments, L is -(CH2) m -C(O)NH-(CH2CH2O) n -(CH2)2-C(O)NH-(CH2) t -C(O)- or -(CH2) m -C(O)NH-(CH2) t -C(O)-, wherein t, m, n are as defined in any embodiment herein.

[0055] In certain embodiments, L is -(CH2) m -C(O)NH-(CH2) t -C(O)-, wherein t, m are as defined in any embodiment herein.

[0056] In certain embodiments, R7is hydrogen or C 1-4alkyl. In certain embodiments, R7is hydrogen, methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl, t-butyl. In certain embodiments, R7is hydrogen, methyl, ethyl, or n-propyl. In certain embodiments, R7is hydrogen, methyl, or ethyl. In certain embodiments, R7is hydrogen.

[0057] In certain embodiments, R8is hydrogen or C 1-4 alkyl. In certain embodiments, R8is hydrogen, methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl, t-butyl. In certain embodiments, R8is hydrogen, methyl, ethyl, or n-propyl. In certain embodiments, R8is hydrogen, methyl, or ethyl. In certain embodiments, R8is hydrogen.

[0058] In certain embodiments, r is 0, 1, 2, or 3. In certain embodiments, r is 0, 1, or 2. In certain embodiments, r is 0 or 1.

[0059] In certain embodiments, p is 0, 1, 2, or 3. In certain embodiments, p is 0, 1, or 2. In certain embodiments, p is 0 or 1.

[0060] In certain embodiments, R6is hydrogen, fluoro, chloro, bromo, iodo, methoxy, ethoxy, propoxy, butoxy, methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl, or t-butyl. In certain embodiments, R6is hydrogen, fluoro, chloro, bromo, iodo, methoxy, ethoxy, propoxy, methyl, ethyl, n-propyl, or i-propyl. In certain embodiments, R6is hydrogen, fluoro, chloro, bromo, methoxy, ethoxy, methyl, or ethyl. In certain embodiments, R6is hydrogen.

[0061] In certain embodiments, t is 1, 2, or 3. In certain embodiments, t is 1 or 2.

[0062] In certain embodiments, m is 1, 2, 3, 4, 5, 6, 7, or 8. In certain embodiments, m is 2, 3, 4, 5, 6, 7, or 8. In certain embodiments, m is 4, 5, or 6. In certain embodiments, m is 7 or 8.

[0063] In certain embodiments, n is 1, 2, 3, 4, 5, 6, 7, or 8. In certain embodiments, n is 2, 3, 4, 5, 6, 7, or 8. In certain embodiments, n is 4, 5, or 6. In certain embodiments, n is 7 or 8.

[0064] In certain embodiments, the compound is selected from:

[0065]

[0066] wherein m, n and t are as defined in any embodiment of the first aspect of the application.

[0067] In certain embodiments, the compound is selected from:

[0068]

[0069]

[0070] The compound of the first aspect of the application can be used in the preparation of an antibody conjugate drug which is capable of specific cleavage to release the active drug under the condition of quinone oxidoreductase 1 (NQOl) and is capable of remaining stable in the absence of NQOl. Accordingly, the second aspect of the application provides the use of a compound of any embodiment of the first aspect of the application, or a racemate, a tautomer, a stereoisomer, an enantiomer, a diastereomer, or a mixture of the aforesaid forms of the compound, or a pharmaceutically acceptable salt or solvate or hydrate thereof, in the preparation of an antibody conjugate drug.

[0071] Linker-active compound conjugates

[0072] The third aspect of the application provides a linker-active compound conjugate, or a racemate, a tautomer, a stereoisomer, an enantiomer, a diastereomer, or a mixture of the aforesaid forms of the linker-active compound conjugate, or a pharmaceutically acceptable salt or solvate or hydrate thereof, which is obtained from the coupling reaction of a compound of any embodiment of the first aspect of the application, or a racemate, a tautomer, a stereoisomer, an enantiomer, a diastereomer, or a mixture of the aforesaid forms of the compound, or a pharmaceutically acceptable salt or solvate or hydrate thereof, with an active compound.

[0073] In certain embodiments, the coupling reaction is a carbamate coupling or a quaternary ammonium salt coupling formed by coupling of an N atom in the molecule of the active compound with the R3 group in the compound of any embodiment of the first aspect of the application.

[0074] In certain embodiments, the conjugate is of formula II,

[0075]

[0076] wherein R1, R2, X1, Z, R5, r, X2, L, R4 are as defined in any embodiment of the application;

[0077] Y is -OC(O)- or is absent;

[0078] B is an active compound.

[0079] In certain embodiments, B is selected from a drug, a cytotoxin, a detection agent, a diagnostic agent or a targeting vector.

[0080] In certain embodiments, B is a cytotoxin, an antitumor drug, an antiviral drug, an anti-infective drug or an immunomodulatory drug.

[0081] In certain embodiments, B is a cytotoxin or an anti-infective drug, such as a tubulin inhibitor, an antitubercular drug, a DNA alkylating agent, a DNA intercalator, an enzyme inhibitor, an antimetabolite, a peptide or nucleotide, a quinolone antibiotic, a rifamycin antibiotic.

[0082] In certain embodiments, B is selected from the group consisting of: an auristatin, monomethyl auristatin E (MMAE), a maytansine or derivative thereof (e.g. a maytansinoid, DM1, DM3, DM4), paclitaxel, a calicheamicin, a duocarmycin, doxorubicin, a camptothecin, a PBD (pyrolobenzodiazepines) cytotoxin and derivative thereof, a rifamycin and derivative thereof, a quinolone antibiotic, the antitubercular drug bedaquiline, the antitubercular drug delamanid, the antitubercular drug Q-203, the antitubercular drug PBTZ-169, KRM-1657.

[0083] In certain embodiments, B is monomethyl auristatin E (MMAE), a rifamycin and derivative thereof, a quinolone antibiotic, the antitubercular drug bedaquiline, the antitubercular drug delamanid, the antitubercular drug Q-203, the antitubercular drug PBTZ-169 or KRM-1657.

[0084] In certain embodiments, the conjugate is selected from the group consisting of:

[0085]

[0086] wherein m, n and t are as defined in any embodiment herein and B is as defined in any embodiment herein.

[0087] In certain embodiments, the conjugate is selected from the group consisting of:

[0088]

[0089] The fourth aspect of the present application provides use of the conjugate of the third aspect of the present application, or a racemate, a tautomer, a stereoisomer, an enantiomer, a diastereomer, or a mixture of the foregoing forms of the present application, or a pharmaceutically acceptable salt or solvate or hydrate thereof, in the manufacture of a medicament for the treatment of a disease.

[0090] Antibody conjugated drugs

[0091] In some embodiments, the antibody conjugate drug is of Formula III,

[0092] In some embodiments, the antibody conjugate drug is of Formula III,

[0093]

[0094] wherein R1, R2, X1, Z, Y, R5, r, X2, L, B are as defined in any embodiment of the present application;

[0095] A is a targeting compound selected from the group consisting of a protein, an antibody, a polypeptide, an enzyme, and a small molecule;

[0096] b represents DAR, which is an integer or a decimal number between 1 and 8.

[0097] In some embodiments, A is an antibody or an antigen-binding fragment thereof.

[0098] In some embodiments, the antibody or antigen-binding fragment thereof comprises:

[0099] (a) the following 3 heavy chain variable region (VH) complementarity determining regions (CDRs):

[0100] (i) a VH CDR1 that has the sequence of CDR1 contained in the VH of SEQ ID NO: 1, or a sequence that has one or several (e.g., 1 or 2 or 3) amino acid substitutions, deletions, or additions compared with the sequence of CDR1 contained in the VH of SEQ ID NO: 1;

[0101] (ii) a VH CDR2 that has the sequence of CDR2 contained in the VH of SEQ ID NO: 1, or a sequence that has one or several (e.g., 1 or 2) amino acid substitutions, deletions, or additions compared with the sequence of CDR2 contained in the VH of SEQ ID NO: 1; and

[0102] (iii) a VH CDR3 that has the sequence of CDR3 contained in the VH of SEQ ID NO: 1, or a sequence that has one or several (e.g., 1) amino acid substitutions, deletions, or additions compared with the sequence of CDR3 contained in the VH of SEQ ID NO: 1;

[0103] and / or

[0104] (b) the following 3 light chain variable region (VL) CDRs:

[0105] (iv) a VL CDR1 that has the sequence of CDR1 contained in the VL of SEQ ID NO: 2, or a sequence of CDR1 contained in the VL of SEQ ID NO: 2 that has one or a few (e.g., 1 or 2) amino acid substitutions, deletions, or additions compared to the sequence of CDR1 contained in the VL of SEQ ID NO: 2;

[0106] (v) a VL CDR2 that has the sequence of CDR2 contained in the VL of SEQ ID NO: 2, or a sequence of CDR2 contained in the VL of SEQ ID NO: 2 that has one or a few (e.g., 1 or 2) amino acid substitutions, deletions, or additions compared to the sequence of CDR2 contained in the VL of SEQ ID NO: 2; and

[0107] (vi) a VL CDR3 that has the sequence of CDR3 contained in the VL of SEQ ID NO: 2, or a sequence of CDR3 contained in the VL of SEQ ID NO: 2 that has one or a few (e.g., 1 or 2) amino acid substitutions, deletions, or additions compared to the sequence of CDR3 contained in the VL of SEQ ID NO: 2.

[0108] In certain embodiments, the antibody or antigen-binding fragment thereof comprises:

[0109] (a) the following 3 heavy chain variable region (VH) complementarity determining regions (CDRs):

[0110] (i) a VH CDR1 that has the sequence of CDR1 contained in the VH of SEQ ID NO: 11, or a sequence of CDR1 contained in the VH of SEQ ID NO: 11 that has one or a few (e.g., 1 or 2 or 3) amino acid substitutions, deletions, or additions compared to the sequence of CDR1 contained in the VH of SEQ ID NO: 11;

[0111] (ii) a VH CDR2 that has the sequence of CDR2 contained in the VH of SEQ ID NO: 11, or a sequence of CDR2 contained in the VH of SEQ ID NO: 11 that has one or a few (e.g., 1 or 2) amino acid substitutions, deletions, or additions compared to the sequence of CDR2 contained in the VH of SEQ ID NO: 11; and

[0112] (iii) a VH CDR3 that has the sequence of CDR3 contained in the VH of SEQ ID NO: 11, or a sequence of CDR3 contained in the VH of SEQ ID NO: 11 that has one or a few (e.g., 1) amino acid substitutions, deletions, or additions compared to the sequence of CDR3 contained in the VH of SEQ ID NO: 11;

[0113] and / or

[0114] (b) the following 3 light chain variable region (VL) CDRs:

[0115] (iv) a VL CDR1 that has the sequence of CDR1 contained in the VL of SEQ ID NO: 12, or a sequence that has one or several (e.g., 1 or 2) amino acid substitutions, deletions, or additions compared to the sequence of CDR1 contained in the VL of SEQ ID NO: 12;

[0116] (v) a VL CDR2 that has the sequence of CDR2 contained in the VL of SEQ ID NO: 12, or a sequence that has one or several (e.g., 1 or 2) amino acid substitutions, deletions, or additions compared to the sequence of CDR2 contained in the VL of SEQ ID NO: 12; and

[0117] (vi) a VL CDR3 that has the sequence of CDR3 contained in the VL of SEQ ID NO: 12, or a sequence that has one or several (e.g., 1 or 2) amino acid substitutions, deletions, or additions compared to the sequence of CDR3 contained in the VL of SEQ ID NO: 12.

[0118] In certain embodiments, the antibody or antigen-binding fragment thereof comprises:

[0119] (a) the following 3 heavy chain variable region (VH) complementarity determining regions (CDRs):

[0120] (i) a VH CDR1 that has the sequence of CDR1 contained in the VH of SEQ ID NO: 21, or a sequence that has one or several (e.g., 1 or 2 or 3) amino acid substitutions, deletions, or additions compared to the sequence of CDR1 contained in the VH of SEQ ID NO: 21;

[0121] (ii) a VH CDR2 that has the sequence of CDR2 contained in the VH of SEQ ID NO: 21, or a sequence that has one or several (e.g., 1 or 2) amino acid substitutions, deletions, or additions compared to the sequence of CDR2 contained in the VH of SEQ ID NO: 21; and

[0122] (iii) a VH CDR3 that has the sequence of CDR3 contained in the VH of SEQ ID NO: 21, or a sequence that has one or several (e.g., 1) amino acid substitutions, deletions, or additions compared to the sequence of CDR3 contained in the VH of SEQ ID NO: 21;

[0123] and / or

[0124] (b) the following 3 light chain variable region (VL) CDRs:

[0125] (iv) a VL CDR1 that has the sequence of CDR1 contained in the VL of SEQ ID NO: 22 or a sequence that has one or several (e.g., 1 or 2) amino acid substitutions, deletions, or additions compared to the sequence of CDR1 contained in the VL of SEQ ID NO: 22;

[0126] (v) a VL CDR2 that has the sequence of CDR2 contained in the VL of SEQ ID NO: 22 or a sequence that has one or several (e.g., 1 or 2) amino acid substitutions, deletions, or additions compared to the sequence of CDR2 contained in the VL of SEQ ID NO: 22; and

[0127] (vi) a VL CDR3 that has the sequence of CDR3 contained in the VL of SEQ ID NO: 22 or a sequence that has one or several (e.g., 1 or 2) amino acid substitutions, deletions, or additions compared to the sequence of CDR3 contained in the VL of SEQ ID NO: 22.

[0128] In certain embodiments, the substitutions are conservative substitutions.

[0129] In certain embodiments, the CDR1, CDR2, and CDR3 contained in the heavy chain variable region (VH) and / or the CDR1, CDR2, and CDR3 contained in the light chain variable region (VL) are defined by the Kabat, Chothia, or IMGT numbering system.

[0130] In certain embodiments, the antibody or antigen-binding fragment thereof comprises:

[0131] the sequence of CDR1, CDR2, and CDR3 contained in the VH of SEQ ID NO: 1; and / or

[0132] the sequence of CDR1, CDR2, and CDR3 contained in the VL of SEQ ID NO: 2.

[0133] In certain embodiments, the antibody or antigen-binding fragment thereof comprises:

[0134] the sequence of CDR1, CDR2, and CDR3 contained in the VH of SEQ ID NO: 11; and / or

[0135] the sequence of CDR1, CDR2, and CDR3 contained in the VL of SEQ ID NO: 12.

[0136] In certain embodiments, the antibody or antigen-binding fragment thereof comprises:

[0137] the sequences of CDR1, CDR2, and CDR3 contained in the VH as shown in SEQ ID NO: 21; and / or

[0138] the sequences of CDR1, CDR2, and CDR3 contained in the VL as shown in SEQ ID NO: 22.

[0139] In certain embodiments, the CDR1, CDR2, and CDR3 contained in the heavy chain variable region (VH), and / or the CDR1, CDR2, and CDR3 contained in the light chain variable region (VL) are defined by the Kabat, Chothia, or IMGT numbering system. In certain embodiments, the CDR1, CDR2, and CDR3 contained in the heavy chain variable region (VH), and / or the CDR1, CDR2, and CDR3 contained in the light chain variable region (VL) are defined by the Kabat numbering system. In certain embodiments, the antibody or antigen-binding fragment thereof comprises:

[0140] (a) the following 3 heavy chain variable region (VH) CDRs:

[0141] (i) a VH CDR1 consisting of the sequence set forth in SEQ ID NO: 3,

[0142] (ii) a VH CDR2 consisting of the sequence set forth in SEQ ID NO: 4, and

[0143] (iii) a VH CDR3 consisting of the sequence set forth in SEQ ID NO: 5;

[0144] and / or

[0145] (b) the following 3 light chain variable region (VL) CDRs:

[0146] (iv) a VL CDR1 consisting of the sequence set forth in SEQ ID NO: 6,

[0147] (v) a VL CDR2 consisting of the sequence set forth in SEQ ID NO: 7, and (vi) a VL CDR3 consisting of the sequence set forth in SEQ ID NO: 8.

[0148] In certain embodiments, the antibody or antigen-binding fragment thereof comprises:

[0149] (a) the following 3 heavy chain variable region (VH) CDRs:

[0150] (i) a VH CDR1 consisting of the sequence set forth in SEQ ID NO: 13,

[0151] (ii) a VH CDR2 consisting of the sequence set forth in SEQ ID NO: 14, and

[0152] (iii) a VH CDR3 consisting of the sequence set forth in SEQ ID NO: 15;

[0153] and / or

[0154] (b) the following 3 light chain variable region (VL) CDRs:

[0155] (iv) a VL CDR1 consisting of the sequence set forth in SEQ ID NO: 16,

[0156] (v) a VL CDR2 consisting of the sequence set forth in SEQ ID NO: 17, and (vi) a VL CDR3 consisting of the sequence set forth in SEQ ID NO: 18.

[0157] In certain embodiments, the antibody or antigen-binding fragment thereof comprises:

[0158] (a) the following 3 heavy chain variable region (VH) CDRs:

[0159] (i) a VH CDR1 consisting of the sequence set forth in SEQ ID NO: 23,

[0160] (ii) a VH CDR2 consisting of the sequence set forth in SEQ ID NO: 24, and

[0161] (iii) a VH CDR3 consisting of the sequence set forth in SEQ ID NO: 25;

[0162] and / or

[0163] (b) the following 3 light chain variable region (VL) CDRs:

[0164] (iv) a VL CDR1 consisting of the sequence set forth in SEQ ID NO: 26,

[0165] (v) a VL CDR2 consisting of the sequence set forth in SEQ ID NO: 27, and (vi) a VL CDR3 consisting of the sequence set forth in SEQ ID NO: 28.

[0166] In certain embodiments, the VH of the antibody or antigen-binding fragment thereof comprises a VH CDR1 as set forth in SEQ ID NO: 3; a VH CDR2 as set forth in SEQ ID NO: 4; and, a VH CDR3 as set forth in SEQ ID NO: 5; and / or, the VL of the antibody or antigen-binding fragment thereof comprises a VL CDR1 as set forth in SEQ ID NO: 6; a VL CDR2 as set forth in SEQ ID NO: 7; and, a VL CDR3 as set forth in SEQ ID NO: 8.

[0167] In certain embodiments, the VH of the antibody or antigen-binding fragment thereof comprises a VH CDR1 as set forth in SEQ ID NO: 13; a VH CDR2 as set forth in SEQ ID NO: 14; and, a VH CDR3 as set forth in SEQ ID NO: 15; and / or, the VL of the antibody or antigen-binding fragment thereof comprises a VL CDR1 as set forth in SEQ ID NO: 16; a VL CDR2 as set forth in SEQ ID NO: 17; and, a VL CDR3 as set forth in SEQ ID NO: 18.

[0168] In certain embodiments, the VH of the antibody or antigen-binding fragment thereof comprises a VH CDR1 as set forth in SEQ ID NO: 23; a VH CDR2 as set forth in SEQ ID NO: 24; and, a VH CDR3 as set forth in SEQ ID NO: 25; and / or, the VL of the antibody or antigen-binding fragment thereof comprises a VL CDR1 as set forth in SEQ ID NO: 26; a VL CDR2 as set forth in SEQ ID NO: 27; and, a VL CDR3 as set forth in SEQ ID NO: 28.

[0169] In certain embodiments, the VH of the antibody or antigen-binding fragment thereof comprises a VH CDR1 as set forth in SEQ ID NO: 3; a VH CDR2 as set forth in SEQ ID NO: 4; and, a VH CDR3 as set forth in SEQ ID NO: 5; and, the VL of the antibody or antigen-binding fragment thereof comprises a VL CDR1 as set forth in SEQ ID NO: 6; a VL CDR2 as set forth in SEQ ID NO: 7; and, a VL CDR3 as set forth in SEQ ID NO: 8.

[0170] In certain embodiments, the VH of the antibody or antigen-binding fragment thereof comprises a VH CDR1 as set forth in SEQ ID NO: 13; a VH CDR2 as set forth in SEQ ID NO: 14; and, a VH CDR3 as set forth in SEQ ID NO: 15; and, the VL of the antibody or antigen-binding fragment thereof comprises a VL CDR1 as set forth in SEQ ID NO: 16; a VL CDR2 as set forth in SEQ ID NO: 17; and, a VL CDR3 as set forth in SEQ ID NO: 18.

[0171] In certain embodiments, the VH of the antibody or antigen-binding fragment thereof comprises a VH CDR1 as set forth in SEQ ID NO: 23; a VH CDR2 as set forth in SEQ ID NO: 24; and, a VH CDR3 as set forth in SEQ ID NO: 25; and, the VL of the antibody or antigen-binding fragment thereof comprises a VL CDR1 as set forth in SEQ ID NO: 26; a VL CDR2 as set forth in SEQ ID NO: 27; and, a VL CDR3 as set forth in SEQ ID NO: 28.

[0172] In certain embodiments, the antibody or antigen-binding fragment thereof comprises:

[0173] (a) a heavy chain variable region (VH) comprising an amino acid sequence selected from the group consisting of:

[0174] (i) the sequence set forth in SEQ ID NO: 1 ;

[0175] (ii) a sequence that has one or several (e.g., 1, 2, 3, 4 or 5) amino acid substitutions, deletions, or additions compared to the sequence set forth in SEQ ID NO: 1 ; and

[0176] (iii) a sequence that has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence set forth in SEQ ID NO: 1 ;

[0177] and

[0178] (b) a light chain variable region (VL) comprising an amino acid sequence selected from the group consisting of:

[0179] (iv) the sequence set forth in SEQ ID NO: 2;

[0180] (v) a sequence that differs from the sequence set forth in SEQ ID NO: 2 by one or several substitutions, deletions, or additions of amino acids (e.g., 1, 2, 3, 4, or 5 substitutions, deletions, or additions of amino acids); and

[0181] (vi) a sequence that has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence set forth in SEQ ID NO: 2.

[0182] In certain embodiments, the antibody or antigen-binding fragment thereof comprises:

[0183] (a) a heavy chain variable region (VH) comprising an amino acid sequence selected from the group consisting of:

[0184] (i) the sequence set forth in SEQ ID NO: 11;

[0185] (ii) a sequence that differs from the sequence set forth in SEQ ID NO: 11 by one or several substitutions, deletions, or additions of amino acids (e.g., 1, 2, 3, 4, or 5 substitutions, deletions, or additions of amino acids); and

[0186] (iii) a sequence that has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence set forth in SEQ ID NO: 11;

[0187] and

[0188] (b) a light chain variable region (VL) comprising an amino acid sequence selected from the group consisting of:

[0189] (iv) the sequence set forth in SEQ ID NO: 12;

[0190] (v) a sequence that differs from the sequence set forth in SEQ ID NO: 12 by one or several substitutions, deletions, or additions of amino acids (e.g., 1, 2, 3, 4, or 5 substitutions, deletions, or additions of amino acids); and

[0191] (vi) a sequence that has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence set forth in SEQ ID NO: 12.

[0192] In certain embodiments, the antibody or antigen-binding fragment thereof comprises:

[0193] (a) a heavy chain variable region (VH) comprising an amino acid sequence selected from the group consisting of:

[0194] (i) the sequence set forth in SEQ ID NO: 21;

[0195] (ii) a sequence that differs from the sequence set forth in SEQ ID NO: 21 by one or several amino acid substitutions, deletions or additions (e.g., 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions); and

[0196] (iii) a sequence that has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence set forth in SEQ ID NO: 21;

[0197] and

[0198] (b) a light chain variable region (VL) comprising an amino acid sequence selected from the group consisting of:

[0199] (iv) the sequence set forth in SEQ ID NO: 22;

[0200] (v) a sequence that differs from the sequence set forth in SEQ ID NO: 22 by one or several amino acid substitutions, deletions or additions (e.g., 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions); and

[0201] (vi) a sequence that has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence set forth in SEQ ID NO: 22.

[0202] In certain embodiments, the substitution is a conservative substitution.

[0203] In certain embodiments, the antibody or antigen-binding fragment thereof comprises:

[0204] a heavy chain variable region (VH) comprising a sequence having the sequence set forth in SEQ ID NO: 1 or having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence set forth in SEQ ID NO: 1, and a light chain variable region (VL) comprising a sequence having the sequence set forth in SEQ ID NO: 2 or having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence set forth in SEQ ID NO: 2.

[0205] In certain embodiments, the antibody or antigen-binding fragment thereof comprises:

[0206] a heavy chain variable region (VH) comprising a sequence having the sequence set forth in SEQ ID NO: 11 or having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence set forth in SEQ ID NO: 11, and a light chain variable region (VL) comprising a sequence having the sequence set forth in SEQ ID NO: 12 or having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence set forth in SEQ ID NO: 12.

[0207] In certain embodiments, the antibody or antigen-binding fragment thereof comprises:

[0208] a heavy chain variable region (VH) comprising a sequence having the sequence set forth in SEQ ID NO: 21 or having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence set forth in SEQ ID NO: 21, and a light chain variable region (VL) comprising a sequence having the sequence set forth in SEQ ID NO: 22 or having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence set forth in SEQ ID NO: 22.

[0209] In certain embodiments, the antibody or antigen-binding fragment thereof comprises a VH having the sequence set forth in SEQ ID NO: 1 and a VL having the sequence set forth in SEQ ID NO: 2.

[0210] In certain embodiments, the antibody or antigen-binding fragment thereof comprises a VH having the sequence set forth in SEQ ID NO: 1 and a VL having the sequence set forth in SEQ ID NO: 2.

[0211] In certain embodiments, the antibody or antigen-binding fragment thereof comprises a VH having the sequence set forth in SEQ ID NO: 1 and a VL having the sequence set forth in SEQ ID NO: 2.

[0212] In certain embodiments, the antibody or antigen-binding fragment thereof comprises a VH having the sequence set forth in SEQ ID NO: 1 and a VL having the sequence set forth in SEQ ID NO: 2.

[0213] In certain embodiments, the antibody or antigen-binding fragment thereof is humanized.

[0214] In certain embodiments, the antibody or antigen-binding fragment thereof further comprises a framework region of a human immunoglobulin.

[0215] In certain embodiments, the antibody or antigen-binding fragment thereof further comprises a heavy chain framework region of a human immunoglobulin (e.g., a heavy chain framework region comprised in an amino acid sequence encoded by a human heavy chain germline antibody gene) and / or, a light chain framework region of a human immunoglobulin (e.g., a light chain framework region comprised in an amino acid sequence encoded by a human light chain germline antibody gene).

[0216] In certain embodiments, the heavy chain framework region and / or light chain framework region optionally comprises one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) back mutations from a human-derived residue to a murine-derived residue.

[0217] In certain embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain having the sequence set forth in SEQ ID NO: 9 and a light chain having the sequence set forth in SEQ ID NO: 10.

[0218] In certain embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain having the sequence set forth in SEQ ID NO: 19 and a light chain having the sequence set forth in SEQ ID NO: 20.

[0219] In certain embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain having a sequence as set forth in SEQ ID NO: 29 and a light chain having a sequence as set forth in SEQ ID NO: 30.

[0220] In certain embodiments, the antibody or antigen-binding fragment thereof further comprises a constant region derived from a human immunoglobulin.

[0221] In certain embodiments, the heavy chain of the antibody or antigen-binding fragment thereof comprises a heavy chain constant region derived from a human immunoglobulin (e.g., IgGl, IgG2, IgG3, IgG4, IgAl, IgA2, IgM, IgD, or IgE).

[0222] In certain embodiments, the light chain of the antibody or antigen-binding fragment thereof comprises a light chain constant region derived from a human immunoglobulin (e.g., kappa or lambda).

[0223] In certain embodiments, the antibody or antigen-binding fragment thereof is selected from the group consisting of:

[0224] (1) a murine-derived antibody, a chimeric antibody (e.g., a human murine chimeric antibody), a humanized antibody, a fully human antibody;

[0225] (2) a full-length antibody, a single chain antibody, a Fab, a Fab', a F(ab')2, a Fv, a scFv, a sdAb, a dAb, a complementarity determining region fragment, a Fd;

[0226] (3) a monoclonal antibody, a polyclonal antibody; and

[0227] (4) a monospecific antibody, a bispecific antibody, a multispecific antibody.

[0228] In certain embodiments, the monoclonal antibody includes a non-CDR region, and the non-CDR region is from a species other than murine, e.g., from a human antibody.

[0229] In certain embodiments, the antibody or antigen-binding fragment thereof is a full-length antibody.

[0230] In certain embodiments, the antibody or antigen-binding fragment thereof is trastuzumab (HERCEPTIN), cetuximab (ERBITUX), bevacizumab (AVASTIN), an anti-tuberculosis antibody, an anti- Staphylococcus aureus antibody, or an antigen-binding fragment thereof.

[0231] In certain embodiments, A is coupled to site # via a S atom or a N atom in the targeting compound molecule.

[0232] In certain embodiments, b is an integer or a decimal number between 2 and 7.

[0233] In certain embodiments, b is an integer or decimal number between 3 and 6 or an integer or decimal number between 4 and 5.

[0234] In certain embodiments, b is an integer or decimal number between 3 and 4.

[0235] In certain embodiments, b is about 2, about 3, about 4, about 5, about 6.

[0236] In certain embodiments, b is an integer or decimal number between 3 and 3.6.

[0237] In certain embodiments, b is about 3.1, about 3.2, about 3.3, about 3.4, or about 3.5.

[0238] In certain embodiments, the antibody drug conjugate is selected from the group consisting of:

[0239]

[0240]

[0241] wherein A, m, n, t, B, b are defined as described in any embodiment of the present application. In certain embodiments, the antibody drug conjugate is selected from the group consisting of:

[0242]

[0243] wherein A, B, b are defined as described in any embodiment of the present application.

[0244] In certain embodiments, the antibody drug conjugate is selected from the group consisting of:

[0245]

[0246]

[0247] wherein A, b are defined as described in any embodiment of the present application.

[0248] Pharmaceutical compositions

[0249] A sixth aspect of the present application provides a pharmaceutical composition comprising at least one antibody drug conjugate of the fifth aspect of the present application, or a racemate, a tautomer, a stereoisomer, an enantiomer, a diastereomer, or a mixture of the foregoing forms, or a pharmaceutically acceptable salt or solvate or hydrate thereof, and one or more pharmaceutically acceptable carriers or excipients.

[0250] In certain embodiments, the antibody conjugate drug of the fifth aspect of the present application, or a racemate, a tautomer, a stereoisomer, an enantiomer, a diastereomer, or a mixture of the foregoing, or a pharmaceutically acceptable salt or solvate or hydrate thereof, is present in the pharmaceutical composition in a therapeutically effective amount.

[0251] In certain embodiments, the pharmaceutical composition can be administered by, for example, oral or parenteral, such as intravenous, intraperitoneal, intramuscular, subcutaneous, etc.

[0252] Use

[0253] The seventh aspect of the present application provides use of the antibody conjugate drug of the fifth aspect of the present application, or a racemate, a tautomer, a stereoisomer, an enantiomer, a diastereomer, or a mixture of the foregoing, or a pharmaceutically acceptable salt or solvate or hydrate thereof, in the manufacture of a medicament, wherein the medicament is for treating or lessening the severity of a disease or condition in a subject

[0254] The eighth aspect of the present application provides a method of treating or lessening the severity of a disease or condition, comprising administering to a patient in need of such treatment a therapeutically effective amount of the antibody conjugate drug of the fifth aspect of the present application, or a racemate, a tautomer, a stereoisomer, an enantiomer, a diastereomer, or a mixture of the foregoing, or a pharmaceutically acceptable salt or solvate or hydrate thereof.

[0255] The ninth aspect of the present application provides the antibody conjugate drug of the fifth aspect of the present application, or a racemate, a tautomer, a stereoisomer, an enantiomer, a diastereomer, or a mixture of the foregoing, or a pharmaceutically acceptable salt or solvate or hydrate thereof, for use in treating or lessening the severity of a disease or condition.

[0256] The tenth aspect of the present application provides a pharmaceutical composition for use in treating or lessening the severity of a disease or condition in a subject, comprising at least one antibody conjugate drug of the fifth aspect of the present application, or a racemate, a tautomer, a stereoisomer, an enantiomer, a diastereomer, or a mixture of the foregoing, or a pharmaceutically acceptable salt or solvate or hydrate thereof.

[0257] In certain embodiments, the disease or condition is selected from the group consisting of a tumor, a bacterial infection, an infectious disease, a hematological disease, a metabolic disease, and an inflammation.

[0258] In certain embodiments, the tumor is selected from the group consisting of a carcinoma, a lymphoma, a lymphoid neoplasm, a blastoma, a sarcoma, a leukemia, and a glioma.

[0259] In some embodiments, the infectious disease is a disease caused by an intracellular bacterial infection.

[0260] In some embodiments, the infectious disease is a disease caused by a Mycobacterium tuberculosis or Staphylococcus aureus infection.

[0261] In some embodiments, the bacterial infection is a Mycobacterium tuberculosis infection or a Staphylococcus aureus infection.

[0262] In some embodiments, the cancer is selected from the group consisting of: breast cancer (e.g., HER2-positive breast cancer); squamous cell cancer (e.g., epithelial squamous cell cancer); lung cancer, including small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, and squamous carcinoma of the lung; cancer of the peritoneum; hepatocellular cancer; gastric or stomach cancer; gastrointestinal cancer; adenocarcinoma; glioblastoma; cervical cancer; ovarian cancer; bladder cancer; urethral cancer; hepatoma; intestinal cancer; colon cancer; rectal cancer; colorectal cancer; endometrial or uterine carcinoma; carcinoma of the fallopian tubes; carcinoma of the vulva; carcinoma of the thyroid; carcinoma of the liver; carcinoma of the esophagus; carcinoma of the gall bladder; carcinoma of the kidney; carcinoma of the renal pelvis; carcinoma of the penis; carcinoma of the testis; carcinoma of the prostate; carcinoma of the skin (e.g., melanoma); carcinoma of the brain; carcinoma of the head and neck; a related metastatic tumor; and combinations of the same.

[0263] Preparation method

[0264] The application also provides a method for preparing the antibody conjugate drug or its racemate, tautomer, stereoisomer, enantiomer, diastereomer, or a mixture of the above-mentioned configurations, or a pharmaceutically acceptable salt or solvate or hydrate thereof according to the fifth aspect of the application, comprising: coupling the reduced antibody or antigen-binding fragment thereof with the conjugate according to the third aspect of the application.

[0265] Advantages of the application

[0266] The antibody conjugate drug provided by the application can specifically release the active drug under the condition of NQO1 enzyme, and can remain stable without NQO1 enzyme. Without being bound by theory, the release mechanism is speculated as follows:

[0267]

[0268] The antibody conjugate drug provided by the application not only shows significant in vitro and in vivo efficacy, but also has relatively ideal drug safety.

[0269] Definitions of terms

[0270] In the present application, unless otherwise specified, the scientific and technical terms used herein have the meanings commonly understood by a person skilled in the art. At the same time, in order to better understand the present application, the definitions and explanations of the related terms are provided as follows.

[0271] When the terms "for example," "e.g.," "for instance," "like," "such as," "include," "including," or "comprising" are used in this document, these terms are not to be interpreted as limiting terms, but rather as specifying the features or elements that are included in the document.

[0272] The terms "a" and "an" and "the" and similar referents in the context of describing the application (especially in the context of the following claims) are to be interpreted in an inclusive rather than an exclusive sense unless otherwise noted or clearly contradicted by context. As used herein, the term "includes" means includes but is not limited to, the term "including" means including but not limited to. The notation "X / Y" means X or Y.

[0273] As used herein, the term "and / or," when used in a list of two or more items, means that any one of the listed items can be present or none of the listed items are present. For example, the phrase "A and / or B" is intended to cover A alone, B alone, or A and B together. As used herein, the term "and / or" when used in a list of two or more items, shall be interpreted to mean that at least one of the items in the list is present, and that the other items in the list can or can not be present.

[0274] As used herein, the term "about" generally means within 0.5% - 10% above or below the indicated value, e.g., within 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% above or below the indicated value. Unless otherwise clear from context, all numerical values provided herein are modified by the term "about."

[0275] As used herein, the term "solvate" refers to a compound of the application associated with a solvent molecule. The solvent can be an organic solvent (e.g., methanol, ethanol, propanol, acetonitrile, etc.), for example, a compound of the application can form an ethanolate with ethanol. A compound of the application can also form a hydrate with water. The amount of solvent can be present in stoichiometric or non-stoichiometric amounts.

[0276] As used herein, the term "stereoisomers" refers to isomers that have the same molecular formula but different structures, resulting from the spatial arrangement of atoms or groups in space. In compounds having one or more asymmetric centers, racemates of the compounds, single enantiomers, mixtures of enantiomers, and mixtures of diastereomers can be formed. The term "stereoisomers" includes conformational isomers and configurational isomers, with the latter including primarily cis-trans isomers and optical isomers. Compounds of the application can exist in stereoisomeric forms, and thus all possible stereoisomeric forms are intended to be embraced within the scope of the present application. For example, single enantiomers, single diastereomers or mixtures of the above are within the scope of the present application. When the compounds of the present application contain olefinic double bonds, unless specified otherwise, the compounds can exist as E or Z isomers, as well as mixtures thereof.

[0277] As used herein, the term "tautomer" refers to isomers of a molecule that differ from each other only in the position of a proton. Compounds of the present application, if tautomers exist, can exist in the form of a single tautomer or mixtures thereof, preferably the more stable tautomer predominates.

[0278] As used herein, the term "racemate" includes "racemic mixture", the term "racemic mixture" refers to an equimolar mixture of chiral molecules with their enantiomers; the term "enantiomer" refers to one of a pair of molecules that are mirror images of each other and are not superimposable, a mixture of enantiomers can be separated, for example, under chiral resolution conditions; the term "diastereomer" refers to a stereoisomer that is not an enantiomer, diastereomers are characterized by some difference in their physical properties, a mixture of diastereomers can be separated, for example, under chromatographic or crystallization conditions.

[0279] Unless otherwise indicated, the compounds, conjugates, and antibody conjugate drugs described herein include isotopically-labeled compounds, which are identical to those recited in the formulae but for the inclusion of an atom that is replaced by an atom having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of suitable isotopically-labeled compounds include, but are not limited to, those that contain the following isotopes: hydrogen isotopes such as 2 H, 3 H; carbon isotopes such as 11 C, 13 C and 14 C; chlorine isotopes such as 36 Cl; fluorine isotopes such as 18 F; nitrogen isotopes such as 13 N and 15 N; oxygen isotopes such as 15 O,17 O and 18 O; and sulfur isotopes such as 35 S. In some embodiments, the compounds, conjugates, and antibody-drug conjugates include deuterium-enriched isotopically-labeled forms thereof.

[0280] The endpoints of the ranges and any values disclosed in this application are not limited to the precise values recited as exactly that number. The exact values recited are intended to be approximations of a closer value for the range letting in the application. For values which are less than one, one unit is considered to be 0.0001, 0.001, 0.01 or 0.1 as appropriate. For values which are greater than one, one unit is considered to be 1, 10, 100, or 1000 as appropriate. For numerical ranges that end with "Ch", "Cm" or "Cn", "n" is a number between 1 and 1000 that is a whole number. For numerical ranges that end with "Cm / Cn", "m" and "n" are numbers between 1 and 1000 that are whole numbers. For numerical ranges that end with "Cm / Cn", "m" and "n" are numbers between 1 and 1000 that are whole numbers. The citation of particular values recited in the ranges should not be interpreted as being limiting to a range which is made up entirely of the particular values recited.

[0281] As used in this application, the term "pharmaceutically acceptable" means that the vehicle, carrier, diluent, excipient, and / or salt is compatible with the other ingredients of the formulation and physiologically tolerable to the recipient.

[0282] As used in this application, the term "pharmaceutically acceptable salt" refers to salts that retain the biological effectiveness and properties of the compounds and are not biologically or otherwise undesirable. In many cases, the compounds disclosed herein are capable of forming acid and / or base salts by virtue of the presence of amino and / or carboxyl groups or groups similar thereto. Pharmaceutically acceptable acid addition salts can be formed with inorganic acids and organic acids. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like. Pharmaceutically acceptable base salts are formed from inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts and the like. Particularly preferred are the ammonium, potassium, sodium, calcium, and magnesium salts. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like. Specific examples include isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. Many such salts are known in the art, for example, as described in WO 87 / 05297, Johnston et al., published September 11, 1987 (hereby incorporated by reference in its entirety).

[0283] As used herein, the term "pharmaceutically acceptable carriers and / or excipients" refers to carriers and / or excipients that are compatible, pharmacologically and / or physiologically, with the subject and the active ingredients, which are well known in the art (see, e.g., Remington's Pharmaceutical Sciences. Edited by Gennaro AR, 19th ed. Pennsylvania: Mack Publishing Company, 1995), and include, but are not limited to, pH adjusting agents, surfactants, adjuvants, ionic strength enhancers, diluents, agents to maintain osmotic pressure, agents to retard absorption, preservatives. For example, pH adjusting agents include, but are not limited to, phosphate buffers. Surfactants include, but are not limited to, cationic, anionic or non-ionic surfactants, such as Tween-80. Ionic strength enhancers include, but are not limited to, sodium chloride. Preservatives include, but are not limited to, various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, and the like. Agents to maintain osmotic pressure include, but are not limited to, sugars, NaCl, and the like. Agents to retard absorption include, but are not limited to, monostearate and gelatin. Diluents include, but are not limited to, water, aqueous buffers (such as buffered saline), alcohols and polyols (such as glycerol), and the like. Stabilizers have the meaning commonly understood by those skilled in the art, which are capable of stabilizing the desired activity of the active ingredients in the pharmaceutical, including, but not limited to, sodium glutamate, gelatin, SPGA, sugars (such as sorbitol, mannitol, starch, sucrose, lactose, dextran, or glucose), amino acids (such as glutamic acid, glycine), proteins (such as dried lactoserum, albumin or casein) or their degradation products (such as lactalbumin hydrolysate), and the like.

[0284] As used herein, the abbreviations for the amino acids have the meaning commonly understood by those skilled in the art. The amino acids are generally L-form amino acids, which can be replaced by D-form amino acids that are chemically close to them, as understood by those skilled in the art. For example, Val represents valine, Ala (A) represents alanine, and Glu (E) represents glutamic acid.

[0285] The term "antibody" in the present application is used in the broadest sense and includes various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments so long as they exhibit the desired antigen-binding activity. The antibodies can be antibodies of different isotypes, for example, IgG (e.g., IgG1, IgG2, IgG3, or IgG4 subtypes), IgA1, IgA2, IgD, IgE, or IgM antibodies.

[0286] As used herein, the term "antigen-binding fragment" of an antibody refers to a polypeptide that comprises a fragment of a full-length antibody that retains the ability to specifically bind the same antigen bound by the full-length antibody, and / or competes with the full-length antibody for specific binding to the antigen, which is also referred to as an "antigen-binding portion." See generally, Fundamental Immunology, Ch. 7 (Paul, W., ed., 2nded. Raven Press, N.Y. (1989)), which is incorporated herein by reference in its entirety. Antigen-binding fragments of an antibody can be produced by recombinant DNA techniques or by enzymatic or chemical cleavage of intact antibodies. Non-limiting examples of antigen-binding fragments include Fab, Fab', F(ab')2, Fd, Fv, complementarity determining region (CDR) fragments, scFv, diabodies, single domain antibodies, chimeric antibodies, linear antibodies, nanobodies, pro bodies, and polypeptides that contain at least a portion of an antibody that is sufficient to confer specific antigen binding

[0287] The terms "full-length antibody," "intact antibody," and "whole antibody" are used herein interchangeably to refer to an antibody having a structure substantially similar to a native antibody structure. Full-length antibodies of the present application can be from a single species, e.g., human; can also be chimeric antibodies or humanized antibodies. Full-length antibodies of the present application comprise two antigen binding sites formed by pairs of VHand VL, respectively, which specifically recognize / bind the same antigen.

[0288] The term "variable region" or "variable domain" refers to the domain of an antibody heavy or light chain that is involved in binding the antibody to an antigen. The variable domains of the heavy chain and light chain (VH and VL, respectively) of a native antibody generally have similar structures, with each domain comprising four conserved framework regions (FRs) and complementary determining regions (CDRs). See, e.g., Kindt et al., Kuby Immunology, 6thEd., W.H. Freeman & Co., page 91 (2007). A single VHor VLdomain can be sufficient to confer antigen-binding specificity. Furthermore, antibodies that bind a particular antigen can be isolated using a VH or VL domain from an antibody that binds the antigen, to screen libraries of complementary VL or VH domains, respectively. See, e.g., Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991). As used herein in connection with variable region sequences, "Kabat numbering" refers to the numbering system set forth by Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed., Public Health Service, National Institutes of Health, Bethesda, MD (1991).

[0289] As used herein, the term "complementarity determining region" or "CDR" refers to amino acid residues in the variable region of an antibody that are responsible for antigen binding. There are three CDRs in each of the variable regions of the heavy and light chains, designated CDR1, CDR2, and CDR3. The precise boundaries of these CDRs can be defined according to various numbering systems known in the art, e.g., as defined by the numbering system of Kabat (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991), Chothia (Chothia & Lesk (1987) J. Mol. Biol. 196:901-917; Chothia et al. (1989) Nature 342:878-883), or IMGT (Lefranc et al., Dev. Comparat. Immunol. 27:55-77, 2003). For a given antibody, one of skill in the art will readily be able to identify the CDRs defined by each numbering system. Moreover, the correspondence between different numbering systems is well understood by those skilled in the art (see, e.g., Lefranc et al., Dev. Comparat. Immunol. 27:55-77, 2003).

[0290] The CDRs contained in the antibodies or antigen-binding fragments thereof of the present application can be determined according to various numbering systems known in the art, e.g., the Kabat, Chothia, or IMGT numbering systems. In certain embodiments, the CDRs contained in the antibodies or antigen-binding fragments thereof of the present application are preferably determined by the Kabat numbering system. One of skill in the art will appreciate that numbering can be performed by other well-known CDR definition means, e.g., Contact, Chothia, IMGT, AbM, etc.

[0291] Unless otherwise indicated, CDR residues and other residues in the variable domains (e.g., FR residues) are numbered herein according to Kabat et al., supra.

[0292] As used herein, the term "Fd" means an antibody fragment consisting of a VH and CHI domain; the term "dAb fragment" means an antibody fragment consisting of a VH domain (Ward et al., Nature 341 :544 546 (1989)); the term "Fab fragment" means an antibody fragment consisting of a VL, VH, CL and CHI domain; the term "F(ab')2 fragment" means an antibody fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; the term "Fab' fragment" means the fragment obtained by reducing the disulfide bonds of a F(ab')2 fragment, consisting of an intact light chain and a Fd fragment of a heavy chain (consisting of a VH and CHI domain).

[0293] As used herein, the term "Fv" means an antibody fragment consisting of the VL and VH domains of a single arm of an antibody. The Fv fragment is commonly held to be the minimum antibody fragment that is capable of forming a complete antigen binding site. It is generally considered that the six CDRs confer the antigen binding specificity of an antibody. However, even a single variable region (e.g., an Fd fragment, which contains only three CDRs specific for an antigen) is capable of recognizing and binding antigen, although it may do so at a lower affinity than the entire binding site.

[0294] As used herein, the term "Fc" means an antibody fragment formed by disulfide bonds between the second, third constant regions of the first heavy chain and the second, third constant regions of the second heavy chain of an antibody. The Fc fragment of an antibody has a variety of different functions, but does not participate in antigen binding.

[0295] As used herein, the term "scFv" refers to a single polypeptide chain comprising a VL and a VH domain, wherein the VL and VH are connected by a linker (see, e.g., Bird et al., Science 242:423-426 (1988); Huston et al., Proc. Natl. Acad. Sci. USA 85:5879-5883 (1988); and Pluckthun, in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994)). Such scFv molecules can have the general structure: NH2-VL-linker-VH-COOH or NH2-VH-linker-VL-COOH. Suitable prior art linkers consist of repeating GGGGS amino acid sequences or variants thereof. For example, a linker having the amino acid sequence (GGGGS)4can be used, but variants thereof can also be used (Holliger et al. (1993), Proc. Natl. Acad. Sci. USA 90:6444-6448). Other linkers useful in the present application are described by Alfthan et al. (1995), Protein Eng. 8:725-731, Choi et al. (2001), Eur. J. Immunol. 31:94-106, Hu et al. (1996), Cancer Res. 56:3055-3061, Kipriyanov et al. (1999), J. Mol. Biol. 293:41-56, and Roovers et al. (2001), Cancer Immunol. In some cases, a disulfide bond can also be present between the VHand VLof an scFv. In certain embodiments, the VHand VLdomains can be positioned relative to each other in any suitable arrangement. For example, scFvs comprising NH2-VH-VH-COOH, NH2-VL-VL-COOH.

[0296] As used herein, the term "single-domain antibody (sdAb)" has the meaning generally understood by those skilled in the art, and refers to an antibody fragment consisting of a single monomeric variable antibody domain, such as a single heavy chain variable region, which retains the ability to specifically bind the same antigen to which a full-length antibody binds. Single-domain antibodies are also known as nanobodies.

[0297] Each of the above-described antibody fragments retains the ability to specifically bind the same antigen to which the full-length antibody binds and / or competes with the full-length antibody for specific binding to the antigen.

[0298] Antigen binding fragments of antibodies (e.g., the antibody fragments described above) can be obtained from a given antibody (e.g., an antibody provided herein) using conventional techniques known to those of skill in the art (e.g., recombinant DNA technology or enzymatic or chemical cleavage methods) and screened for specificity in the same manner as is done for whole antibodies.

[0299] As used herein, the term "murine antibody" refers to an antibody obtained by fusing B cells from an immunized mouse with myeloma cells, screening for murine hybridoma cells that both proliferate indefinitely and secrete antibody, followed by screening, antibody production, and antibody purification, or refers to an antibody secreted by a plasma cell that is formed by the differentiation and proliferation of B cells after an antigen invades a mouse.

[0300] As used herein, the term "humanized antibody" refers to a non-human antibody that has been genetically engineered to have an amino acid sequence that is modified to increase homology to the sequence of a human antibody. Typically, a humanized antibody has all or a portion of the CDR regions from a non-human antibody (donor antibody) and all or a portion of the non-CDR regions (e.g., variable region FR and / or constant region) from a human immunoglobulin (acceptor antibody). A humanized antibody typically retains the desired properties of the donor antibody, including but not limited to, antigen specificity, affinity, reactivity, ability to increase immune cell activity, ability to enhance an immune response, and the like. The donor antibody can be a mouse, rat, rabbit, or non-human primate (e.g., cynomolgus monkey) antibody that has the desired properties (e.g., antigen specificity, affinity, reactivity, ability to increase immune cell activity, and / or ability to enhance an immune response).

[0301] As used herein, the term "identity" refers to the relationship between the sequences of two or more polypeptide molecules or two or more nucleic acid molecules, as determined by aligning and comparing the sequences. "Percent identity" refers to the percentage of identical residues between the compared molecules, and is calculated based on the size of the smallest molecule being compared. For these calculations, gaps in the alignment, if any, are addressed by a particular mathematical model or computer program (i.e., "algorithm"). Methods that can be used to calculate the identity of an aligned nucleic acid or polypeptide include those described in Computational Molecular Biology, (Lesk, A.M., ed.), 1988, New York: Oxford University Press; Biocomputing Informatics and Genome Projects, (Smith, D.W., ed.), 1993, New York: Academic Press; Computer Analysis of Sequence Data, Part I, (Griffin, A.M. and Griffin, H.G., eds.), 1994, New Jersey: Humana Press; von Heinje, G., 1987, Sequence Analysis in Molecular Biology, New York: Academic Press; Sequence Analysis Primer, (Gribskov, M. and Devereux, J., eds.), 1991, New York: M. Stockton Press; and Carillo et al., 1988, SIAM J. Applied Math. 48:1073.

[0302] As used herein, the term "conservative substitution" means an amino acid substitution that does not adversely affect or alter the intended properties of a protein / polypeptide comprising the amino acid sequence. For example, conservative substitutions can be introduced by standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis. Conservative amino acid substitutions include substitutions of an amino acid residue with an amino acid residue having a similar side chain, e.g., substitutions made between residues within the same family that are physicochemically or functionally similar (e.g., have similar size, shape, charge, chemical properties, including ability to form covalent or hydrogen bonds, etc.). Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, and histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), beta-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, a preference is given to substituting the corresponding amino acid residue with another amino acid residue from the same side chain family. Methods for identifying amino acid conservative substitutions are well known in the art (see, e.g., Brummell et al., Biochem. 32: 1180-1187 (1993); Kobayashi et al. Protein Eng. 12(10): 879-884 (1999); and Burks et al. Proc. Natl Acad. Set USA 94: 412-417 (1997), which are incorporated herein by reference).

[0303] As used herein, the term "cytotoxin" refers to those molecules that can be toxic to a cancer cell upon release in that cell. Toxins of particular interest in the present application include methyl auristatin E (MMAE), auristatin, maytansinoid or derivatives thereof (e.g. maytansinoid, DM1, DM3, DM4), calicheamicin, duocarmycin, doxorubicin, camptothecin or PBD class cytotoxins. Anti-infective drugs of particular interest are rifamycin derivatives, quinolone antibiotics, anti-tuberculosis drug bedaquiline, anti-tuberculosis drug delamanid, anti-tuberculosis drug Q-203, anti-tuberculosis drug PBTZ-169, KRM-1657 and the like.

[0304] As used herein, the term "linker" is a molecule having two reactive ends, one of which can be coupled to an antibody and the other of which is used to couple to an active compound (e.g., a cytotoxin, an antitumor drug, an antiviral drug, an anti-infective drug, or an immunomodulator drug). The antibody-coupling end of the linker is typically a site capable of coupling to a thiol of a cysteine or an amine of a lysine on the antibody, and the active compound-coupling end of the linker is typically a site capable of coupling to a thiol, an amine, a carboxyl, or a hydroxyl on the active compound molecule. When the term linker is used to describe a coupled form of the linker, it can no longer include one or both of the reactive end sites (e.g., a leaving group for a thiol-reactive group, a leaving group for an amine-reactive group) because the linker has reacted to form a covalent bond with one or both of the antibody and the active compound.

[0305] As used herein, the term "antibody conjugate drug" or "ADC" is a product formed by coupling a plurality (typically 1-8) of active compounds (e.g., cytotoxins, antitumor drugs, antiviral drugs, anti-infective drugs, or immunomodulator drugs) to an antibody molecule via linkers. The antibody conjugated to one or more active compounds is typically a monoclonal antibody selective for a specific antigen (e.g., a cancer or a bacterium).

[0306] As used herein, "b" refers to the drug loading, i.e., the average number of active compounds loaded on each antibody of Formula III, which can also be expressed as the ratio of the amount of drug to the amount of antibody (DAR), and the drug loading can range from 0.1-10, preferably 1-8, 2-7, 3-6 active compounds per antibody (Ab). The number of drug moieties per ADC molecule after the conjugation reaction can be identified using conventional methods such as UV / visible spectroscopy, mass spectrometry, ELISA assays, and HPLC profiling.

[0307] In one embodiment of the application, the active compound is conjugated to the antibody via a linker at a cysteine thiol-SH on an antibody chain that is opened or at a thiol-SH of a cysteine residue that is the result of a site-directed mutation, or the active compound is conjugated to the antibody via a linker at a lysine side chain amine NH2 on the antibody. Generally, the number of drug molecules that can be conjugated to the antibody in a conjugation reaction will be less than or equal to the theoretical maximum.

[0308] As used herein, C l-6 "1 and 6" means the number of carbon atoms in the designated group. That is, the group can include carbon atom numbers from "1" to "6" inclusive, all alkyl groups having from 1 to 6 carbon atoms, including straight-chain or branched-chain alkyl groups. Thus, for example, "C1-C4 alkyl" or "C 1-4"Alkyl" refers to all alkyl groups having from 1 to 4 carbon atoms, i.e., CH3, CH3CH2, CH3CH2CH2-, (CH3)2CH-, CH3CH2CH2CH2-, CH3CH2CH(CH3)-, and (CH3)3C-.

[0309] As used herein, the term "C 1-6 "Alkoxy" refers to a radical of the formula: -O-alkyl, wherein alkyl is as previously described. For example, C1-6alkoxy, C1-4alkoxy, C1-2alkoxy, C3-6cycloalkoxy, C3-6cycloalkyl-C1-2alkoxy, C4-6cycloalkyl-C1-2alkoxy, and the like. 1-6 "Alkyl-O-" refers to a radical of the formula: -O-alkyl, wherein alkyl is as previously described. For example, C1-6alkyl-O-, C1-4alkyl-O-, C1-2alkyl-O-, C3-6cycloalkyl-O-, C3-6cycloalkyl-C1-2alkyl-O-, C4-6cycloalkyl-C1-2alkyl-O-, and the like. 1-6 "Alkyl" has the same meaning as previously described for "C 1-6 "Alkyl" has the same meaning as previously described for "C 1-4 "Alkoxy" refers to a radical of the formula: -O-alkyl, wherein alkyl is as previously described. For example, C1-6alkoxy, C1-4alkoxy, C1-2alkoxy, C3-6cycloalkoxy, C3-6cycloalkyl-C1-2alkoxy, C4-6cycloalkyl-C1-2alkoxy, and the like. 1-2 "Alkoxy" refers to a radical of the formula: -O-alkyl, wherein alkyl is as previously described. For example, C1-6alkoxy, C1-4alkoxy, C1-2alkoxy, C3-6cycloalkoxy, C3-6cycloalkyl-C1-2alkoxy, C4-6cycloalkyl-C1-2alkoxy, and the like. 1-4 "Alkoxy" refers to a radical of the formula: -O-alkyl, wherein alkyl is as previously described. For example, C1-6alkoxy, C1-4alkoxy, C1-2alkoxy, C3-6cycloalkoxy, C3-6cycloalkyl-C1-2alkoxy, C4-6cycloalkyl-C1-2alkoxy, and the like.

[0310] As used herein, the term "halogen" refers to any one of the radioactively stable atoms in column 7 of the Periodic Table of the Elements, e.g., fluorine, chlorine, bromine, iodine, and the like, with chlorine being the preferred.

[0311] "Individual" or "subject" is preferably a mammal. Mammals include, but are not limited to, domesticated animals (e.g., cows, sheep, cats, dogs, and horses), primates (e.g., humans and non-human primates such as monkeys), rabbits, and rodents (e.g., mice and rats). In certain embodiments, the individual or subject is a human.

[0312] As used herein, the term "treatment" is intended to refer to alleviating, abating, ameliorating, or eradicating a disease state or condition being treated. An individual is successfully "treated" if, after receiving a therapeutic amount of the antibody drug conjugate or racemate, tautomer, stereoisomer, enantiomeric, diastereomeric, or mixture of the foregoing, or a pharmaceutically acceptable salt or solvate or hydrate thereof according to the methods described herein, the individual exhibits an observable and / or detectable decrease in one or more signs and symptoms. It will also be appreciated that the treatment of the disease state or condition includes instances where not all signs and symptoms are completely eliminated, but some biologically or medically relevant result is achieved.

[0313] The term "treatment" as used herein in the context of treating a condition generally relates to the treatment and therapy of a human or animal with the purpose to achieve a desired therapeutic effect, such as inhibition of progression of the condition, including a decrease in the rate of progression, stasis of progression, regression of the condition, amelioration of the condition, and cure of the condition. For cancer, "treatment" can refer to inhibition or slowing of tumor or malignant cell growth, proliferation, or metastasis, or some combination thereof.

[0314] The term "effective amount" as used herein relates to the amount of an active compound or the amount of a material, composition, or dosage form containing an active compound, which is effective for producing some desired therapeutic effect in accordance with the treatment regimen intended. For example, when used in conjunction with treating a disease or disorder, "effective amount" refers to the amount or concentration of an antibody conjugate drug or a racemate, tautomer, stereoisomer, enantiomer, diastereomer, or mixture of the foregoing forms, or a pharmaceutically acceptable salt or solvate or hydrate thereof, effective to treat the disease or disorder.

[0315] It is further noted that the dosage and method of use of the antibody conjugate drug of the present application is dependent on a number of factors, including age, body weight, sex, natural health status, nutritional status, activity of the compound, time of administration, metabolic rate, severity of the condition, and the judgment of the attending medical or veterinary practitioner. Preferred dosages are between 0.001 and 1000 mg / kg body weight per day.

[0316] Sequence information

[0317] Information of the partial sequences involved in the present application is shown in the following table:

[0318]

[0319]

[0320] DETAILED DESCRIPTION

[0321] The essential characteristics of the present application are further illustrated in the following examples, which are intended to be purely exemplary of the present application and not limitative of the scope of the application. In the following examples, unless otherwise indicated, the conditions are conventional conditions or those as recommended by the manufacturer. The materials used in the following examples are commercially available unless otherwise indicated.

[0322] Although many of the materials and procedures used in the following examples are known in the art, the present application will nevertheless describe them in some detail. Those skilled in the art will appreciate that the materials and procedures used in the following examples are described in detail only to the extent necessary to understand the present application, unless specifically noted otherwise.

[0323] Example 1: Synthesis of Antibody Conjugate Drug Key Intermediate M12 Based on Benzoquin Structure Linker

[0324]

[0325] 1) Preparation of M01

[0326] Dissolve 2,6-dimethylhydroquinone (7.74 g, 56.02 mmol) completely with 400 mL of methyl sulfonic acid, add 3,3-dimethylacrylic acid (11.21 g, 112.04 mmol), replace with nitrogen three times after vacuumizing, heat to 70°C and reflux the reaction for 8 h. After the reaction is completed, cool the reaction liquid to room temperature, extract with ethyl acetate, distilled water, saturated sodium bicarbonate solution and saturated brine successively, dry with anhydrous sodium sulfate, evaporate the solvent under reduced pressure, and recrystallize with ethyl acetate-petroleum ether to obtain white solid powder M01 (9.91 g, 81.1% yield). 1 H-NMR (400 MHz, DMSO-d6) δ 8.12 (s, 1H), 6.66 (s, 1H), 2.60 (s, 2H), 2.28 (s, 3H), 2.13 (s, 3H), 1.35 (s, 6H). ESI m / z: calculated for C 13 H 16 O3 220.10; found 221.12 [M+H] + ; 243.11 [M+Na] + .

[0327] 2) Preparation of M02

[0328] Dissolve M01 (9.01 g, 40.95 mmol) in 1000 mL of a flask, then slowly add bromine water (19.65 g, 122.86 mmol) after dissolving in methanol (200 mL), and stir the reaction at room temperature for 24 h. Add saturated sodium thiosulfate solution, and the reaction liquid changes from red-brown to yellow. Add distilled water, extract with dichloromethane successively, wash with saturated brine, and dry with anhydrous sodium sulfate. Evaporate the solvent under reduced pressure, and separate and purify by column chromatography (dichloromethane:methanol = 50:1 to 10:1 gradient elution, V / V) to obtain white solid powder M02 (4.50 g, 56.55% yield). 1 H-NMR (400 MHz, Chloroform-D) δ 4.64 (s, 1H), 2.58 (s, 2H), 2.39 (s, 3H), 2.35 (s, 3H), 1.47 (s, 6H). ESI m / z: calculated for C 13 H 15BrO3298.02; found 299.03 [M+H] + .

[0329] 3) Preparation of M03

[0330] M02(4.95 g, 16.61 mmol) was added to a mixed solution of 250 mL of acetonitrile and 100 mL of water, and N-bromosuccinimide (3.54 g, 19.93 mmol) was added portionwise, and the reaction was stirred at room temperature overnight. After the reaction was completed, it was extracted with ethyl acetate, washed with distilled water and saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a yellow oil M03 (5.03 g, 96.5%). 1 H-NMR (400 MHz, DMSO-d6) δ 12.21 (s, 1H), 2.85 (s, 2H), 2.11 (s, 3H), 2.08 (s, 3H), 1.37 (s, 6H). ESI m / z: calculated for C 13 H 15 BrO4 314.01; found 315.01 [M+H] + .

[0331] 4) Preparation of M04

[0332] M03 (6.05 g, 19.20 mmol) was dissolved in 100 mL of methanol, and then the reaction solution was transferred to a low-temperature reaction tank and cooled to 0°C, and thionyl chloride (3.42 g, 28.80 mmol) was slowly added portionwise dropwise, and after the addition was completed, the reaction solution was transferred to room temperature and stirred overnight. After the reaction was completed, it was concentrated under reduced pressure, and separated and purified by column chromatography (dichloromethane:methanol = 50:1 to 10:1 gradient elution, V / V) to obtain a yellow oil M04 (5.13 g, 81.5%). 1 H-NMR (400 MHz, Chloroform-D) δ 3.60 (s, 3H), 2.98 (s, 2H), 2.18 (d, J = 2.6 Hz, 6H), 1.44 (s, 6H). ESI m / z: calculated for C 14 H 17 BrO4 328.03; found 329.04 [M+H] + ; 351.02 [M+Na] + .

[0333] 5) Preparation of M05

[0334] Sodium azide (3.56 g, 54.74 mmol) was dissolved in 20 mL distilled water, M04 (5.13 g, 15.64 mmol) was dissolved in 200 mL methanol, the sodium azide aqueous solution was added slowly dropwise into the reaction solution, and the reaction was stirred at room temperature for 24 h. After the reaction was completed, the solvent was evaporated under reduced pressure to obtain yellow oily M05 (4.19 g, 92.08%).

[0335] 6) Preparation of M06

[0336] M05 (4.19 g, 14.39 mmol) was dissolved in 200 mL dichloromethane, triphenylphosphine (4.15 g, 15.83 mmol) was added portionwise into the reaction solution, the reaction system changed from light red to dark red, and the reaction was stirred at room temperature overnight. The solvent was evaporated under reduced pressure, 150 mL of tetrahydrofuran, distilled water and glacial acetic acid were added, the temperature was heated to 65°C to reflux the reaction for 8 h. After the reaction was completed, most of the water and acetic acid were evaporated under reduced pressure, the residue was extracted with ethyl acetate, washed with distilled water, saturated sodium bicarbonate solution and saturated brine in turn, dried over anhydrous sodium sulfate, evaporated under reduced pressure, and separated and purified by column chromatography (ethyl acetate: petroleum ether = 1:10 to 1:1 gradient elution, V / V) to obtain dark red solid powder M06 (3.06 g, 80.5%). 1 H-NMR (400 MHz, DMSO-d6) δ 6.36 (s, 2H), 3.51 (s, 3H), 2.91 (s, 2H), 2.09 (s, 3H), 1.73 (s, 3H), 1.37 (s, 6H). ESIm / z: calculated for C 14 H 19 NO4 265.13; found 266.14 [M+H] + ; 288.12 [M+Na] + .

[0337] 7) Preparation of M07

[0338] M06 (3.06 g, 11.54 mmol) was dissolved in a mixed solution of 250 mL ether and 100 mL distilled water, sodium dithionite (6.02 g, 34.62 mmol) was added portionwise into the reaction solution, and the reaction was stirred at room temperature for 8 h, and the reaction system changed from red to colorless. After the reaction was completed, dichloromethane was used for extraction, and distilled water and saturated brine were used for washing in turn, dried over anhydrous sodium sulfate, evaporated under reduced pressure, and recrystallized from ethyl acetate-petroleum ether to obtain white solid powder M07 (2.34 g, 86.34%). 1H-NMR (400 MHz, DMSO-d6) δ 7.77 (s, 1H), 4.41 (s, 2H), 2.59 (s, 2H), 2.19 (s, 3H), 1.98 (s, 3H), 1.34 (s, 6H). ESI m / z: calculated for C 13 H 17 NO3 235.12; found 236.13 [M+H] + ; 258.11 [M+Na] + .

[0339] 8) Preparation of M08

[0340] Boc-glycine (2.09 g, 11.94 mmol) was dissolved in 35 mL of N,N-dimethylformamide, and then the reaction solution was transferred to a low-temperature reaction tank and cooled to 0 °C. N,N-diisopropylethylamine (3.85 g, 29.85 mmol) and HATU (4.54 g, 11.94 mmol) were added, and after stirring at 0 °C for 15 minutes, M07 (2.34 g, 9.95 mmol) was added, and the solution was transferred to room temperature and stirred overnight. After the reaction was completed, distilled water was added to quench the reaction, and ethyl acetate was sequentially extracted, washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. Column chromatography (dichloromethane:methanol = 50:1 to 10:1 gradient elution, V / V) was used to separate and purify the white solid powder M08 (1.30 g, 33.3%). 1 H-NMR (400 MHz, DMSO-d6) δ 9.22 (s, 1H), 8.19 (s, 1H), 7.04 (t, J = 4.0 Hz, 1H), 3.74 (d, J = 4.1 Hz, 2H), 2.58 (s, 2H), 2.31 (s, 3H), 1.97 (s, 3H), 1.40 (s, 9H), 1.37 (s, 6H). ESI m / z: calculated for C 20 H 28 N2O6 392.19; found293.15 [M-boc] - .

[0341] 9) Preparation of M09

[0342] M08 (1.60 g, 4.07 mmol) was dissolved in 20 mL of dichloromethane, and trifluoroacetic acid (2.32 g, 20.35 mmol) was added, and the solution was stirred at room temperature for 2 h. After the reaction was completed, the solvent was evaporated under reduced pressure, and 20 mL of a mixed solution of toluene:dichloromethane (5:1) was added to evaporate the solvent under reduced pressure. After repeating the evaporation three times, a black oily substance was obtained as a trifluoroacetate salt of the target product M09.

[0343] 10) Preparation of M10

[0344] The trifluoroacetate salt of M09 (1.60 g) was dissolved in 25 mL of N,N- dimethylformamide, N,N-diisopropylethylamine (3.15 g, 24.4 mmol) and 6- (maleimidyl)hexanoic acid succinimidyl ester (1.50 g, 4.88 mmol) were added and stirred at room temperature for 18 h. After the reaction was completed, distilled water was added to quench, extracted with dichloromethane, washed with saturated brine in sequence, dried over anhydrous sodium sulfate, evaporated under reduced pressure and separated and purified by column chromatography (dichloromethane:methanol = 50:1 to 20:1 gradient elution, V / V) to obtain white solid powder M10 (0.60 g, 31.5%). 1 H-NMR (400 MHz, DMSO-d6) δ 9.23 (s, 1H), 8.19 (s, 1H), 8.10 (t, J = 3.8 Hz, 1H), 6.99 (s, 2H), 3.87 (d, J = 3.8 Hz, 2H), 3.37 (t, J = 4.7 Hz, 2H), 2.57 (s, 2H), 2.29 (s, 3H), 2.13 (t, J = 5.0 Hz, 2H), 1.95 (s, 3H), 1.52-1.45 (m, 4H), 1.36 (s, 6H), 1.23-1.18 (m, 2H). ESI m / z: calculated for C 25 H 31 N3O7 485.21; found 486.24 [M+H] + ; 508.22 [M+Na] + .

[0345] 11) Preparation of M11

[0346] M10 (220 mg, 0.453 mmol) was dissolved in 10 mL of a mixed solution of acetonitrile, distilled water and acetone (10:10:1), and N-bromosuccinimide (80.2 mg, 0.453 mmol) was completely dissolved using 1 mL of acetonitrile. The reaction solution was transferred to a low-temperature reaction tank and cooled to -5°C, and the acetonitrile solution of N-bromosuccinimide was added dropwise in portions. After the reaction was completed, 200 mL of dichloromethane was used for extraction, washed with saturated brine in sequence, dried over anhydrous sodium sulfate, evaporated under reduced pressure and obtained as a yellow oil M11 (120 mg, 53.1%). 1H-NMR (400 MHz, DMSO-d6) δ 12.04 (s, 1H), 9.62 (s, 1H), 8.12 (t, J = 4.0 Hz, 1H), 6.99 (s, 2H), 3.92 (d, J = 3.9 Hz, 2H), 3.37 (t, J = 4.7 Hz, 2H), 2.80 (s, 2H), 2.13 (t, J = 4.8 Hz, 2H), 2.11 (s, 3H), 1.76 (s, 3H), 1.51 - 1.46 (m, 4H), 1.36 (s, 6H), 1.25 - 1.19 (m, 2H). ESI m / z: calculated for C 25 H 31 N3O8 501.21; found 500.20 [M-H] - .

[0347] 12) Preparation of M12

[0348] M11 (200 mg, 0.399 mmol) was dissolved in 1 ml of dichloromethane, N,N- diisopropylethylamine (69 μl, 0.399 mmol) and isobutyl chloroformate (52 μl, 0.399 mmol) were added portion wise under ice bath, TLC was monitored until the reaction was completed, then p-aminophenol (49 mg, 0.399 mmol) was added and the reaction was allowed to proceed at room temperature overnight. After the reaction was completed, it was washed with water, the organic layer was concentrated and flash column chromatography was performed to obtain M12 as yellow oil 100 mg, 41.5%. 606.26; found 607.28 [M+H] - . 1 H-NMR (400 MHz, DMSO-d6) δ 9.62 (s, 1H), 8.35 (s, 1H), 7.52 (d, J = 6.4 Hz, 2H), 7.26 (d, J = 7.2 Hz, 2H), 7.16 (t, J = 4.8 Hz, 1H), 6.70 (s, 2H), 4.60 (d, J = 4.6 Hz, 2H), 3.94 (d, J = 4.6 Hz, 2H), 3.79 (t, J = 4.8 Hz, 2H), 2.82 (s, 2H), 2.61 (s, 3H), 2.55 (t, J = 4.6 Hz, 1H), 2.23 (t, J = 6.6 Hz, 2H), 2.15 (s, 3H), 1.69 (p, J = 4.7 Hz, 2H), 1.54 - 1.48 (m, 2H), 1.43 (s, 6H), 1.41 - 1.37 (m, 2H).

[0349] Example 2: Preparation of ADC-01 based on benzoquinone structure linker-MMAE

[0350] The equation of the reaction is as follows:

[0351]

[0352] M12 (80 mg, 0.132 mmol), bis(4-nitrophenyl) carbonate (80.3 mg, 0.264 mmol) were dissolved in 10 ml N,N-dimethylformamide, N,N-diisopropylethylamine (23 μl, 0.132 mmol) was added and stirred at room temperature overnight. After the reaction was completed, the solvent was evaporated under reduced pressure, and 10 ml of anhydrous diethyl ether was slurried twice to obtain brown solid M13 58 mg. 57.1 %. 771.27; found 772.28 [M+H] - . 1 H-NMR (400 MHz, DMSO-d6) δ 9.62 (s, 1H), 8.35 (s, 1H), 7.48 (d, J = 6, 2H), 7.36 - 7.30 (m, 8H), 7.16 (t, J = 4.8 Hz, 1H), 6.90 (d, J = 7.0 Hz, 1H), 6.70 (s, 2H), 5.15 (s, 2H), 4.78 - 4.73 (m, 1H), 4.43 - 4.39 (m, 2H), 4.16 (d, J = 4.2 Hz, 1H), 4.14 - 4.11 (m, 1H), 4.05 - 4.00 (m, 2H), 3.95 (d, J = 4.6 Hz, 2H), 3.81 - 3.77 (m, 3H), 3.71 - 3.66 (m, 1H), 3.61 - 3.56 (m, 1H), 3.48 - 3.44 (m, 1H), 3.33 (dd, J = 9.5, 1.0 Hz, 6H), 2.88 (t, J = 1.1 Hz, 6H), 2.82 (s, 2H), 2.73 - 2.62 (m, 2H), 2.61 (s, 2H), 2.53 (dd, J = 12.4, 5.4 Hz, 1H), 2.24 - 2.16 (m, 3H), 2.15 (s, 3H), 2.05 - 2.00 (m, 1H), 1.95 - 1.67 (m, 7H), 1.54 - 1.45 (m, 3H), 1.43 (s, 6H), 1.41 - 1.37 (m, 2H), 1.28 (dd, J = 5.0, 1.2 Hz, 3H), 1.23 - 1.18 (m, 1H), 1.15 (dd, J = 5.6, 1.2 Hz, 1H), 0.94 - 0.80 (m, 18H).

[0353] M13 (57 mg, 0.074 mmol) was dissolved in N,N-dimethylformamide (DMF). Subsequently, 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU, 45.6 mg, 0.137 mmol) and N,N-diisopropylethylamine (DIPEA, 19.37 mg, 0.171 mmol) were introduced into the solution. Thereafter, a DMF solution containing monomethyl auristatin E (MMAE) (72 mg, 0.114 mmol) was added dropwise to the reaction mixture. The resulting mixture was then stirred at ambient temperature for 24 hours. After completion of the reaction, the solvent was removed under reduced pressure. Finally, the reaction product was purified using column chromatography to obtain M14 (100 mg, 73.52%) as a yellow solid.1H NMR (400 MHz, DMSO-d6) δ 9.39-9.23 (m, 1H), 8.60 (d, J = 5.8 Hz, 1H), 8.10 (s, 1H), 7.90 (d, J = 5.8 Hz, 1H), 7.62 (d, J = 5.8 Hz, 1H), 7.31-7.25 (m, 7H), 7.17 (d, J = 5.6 Hz, 1H), 7.00 (s, 2H), 5.40 (s, 1H), 4.90-4.85 (m, 2H), 4.76-4.43 (m, 3H), 4.00-3.78 (m, 5H), 3.58-3.46 (m, 1H), 3.37 (t, J = 4.8 Hz, 2H), 3.24-3.11 (m, 8H), 3.05-2.91 (m, 5H), 2.80-2.69 (m, 2H), 2.41 (t, J = 9.4 Hz, 1H), 2.30-2.25 (m, 2H), 2.12 (d, J = 3.8 Hz, 4H), 2.08-2.04 (m, 3H), 1.95 (s, 1H), 1.74 (s, 6H), 1.48 (s, 7H), 1.36 (s, 6H), 1.23-1.20 (m, 6H), 1.05-0.97 (m, 6H), 0.90-0.75 (m, 16H). HR-MS m / z 1413.877; found 1414.707 [M+H]+, 1436.686 [M+Na]+.

[0354] The intermediate M12 prepared in Example 1 was reacted with MMAE to obtain the linker-MMAE (M14), and M14 was conjugated with the anti-EGFR antibody cetuximab to obtain ADC-01.

[0355] 1) Preparation of common buffer salt solutions:

[0356] Buffer-1: Take 3.11 g of L-histidine, dissolve in 1 L of double distilled water, after completely dissolved, use medical glacial acetic acid to adjust its pH = 5.50 (± 0.05); after filtering bacteria through 0.22 μm filter membrane, bottle and store at 4 °C for short-term storage, and wait for use.

[0357] Buffer-2: Take 6.06 g of TRIS·base and 0.93 g of EDTA·2Na, dissolve, and then make up to 100 mL; take 7.88 g of TRIS·HCl and 0.93 g of EDTA·2Na, dissolve, and then make up to 100 mL; add the TRIS·HCl solution to the TRIS·base solution, and adjust to pH = 8.50 (± 0.05); after filtering bacteria through 0.22 μm filter membrane, bottle and store at 4 °C for short-term storage, and wait for use.

[0358] Buffer-3: Take 1.715 mL of medical glacial acetic acid, dissolve in 200 mL of double distilled water, mix thoroughly, and then filter bacteria through 0.22 μm filter membrane, bottle and store at 4 °C for short-term storage, and wait for use.

[0359] 2) Antibody conjugation reaction:

[0360] 2.1 Displacement of the pharmaceutical antibody: First, freeze-thawed cetuximab stock solution (purchased from Merck Company) is slowly thawed at room temperature, and is displaced into buffer-1 through G25 dextran gel column. After displacement is completed, concentration is performed by ultrafiltration centrifugation (final concentration > 5 mg / mL), and the concentration is determined by ultraviolet spectrophotometer.

[0361] 2.2 Preparation of the conjugation reaction solution: According to the amount of the antibody (1 eq) to be conjugated, the antibody buffer-1 solution is accurately taken by using a pipette, and a certain amount of buffer-1 is added to make the antibody concentration about 10 mg / mL. The pH is adjusted to about 6-8 by using buffer-2, and the solution is transferred to a clean reaction vial by using a pipette.

[0362] 2.3 Reduction of the antibody: The reaction solution in the vial is slowly stirred (100 rpm), and 2-5 eq of 2.87 mg / mL TCEP·HCl solution is added. After addition is completed, the solution is slowly stirred at room temperature for 60-180 min.

[0363] 2.4 Antibody conjugation: Calculate the volume of organic solvent (DMAC or DMSO) needed to be added to make up 5% to 15% of the total volume; at the same time, calculate the mass of ADC small molecule payload (Linker-MMAE) needed to be added, usually a slight excess (usually 8 eq) of small molecule payload is needed, then calculate the concentration of the payload in the organic solvent needed to be added. After accurately preparing the solution of ADC small molecule payload, slowly add it into the reduced antibody reaction solution. Continue to slowly stir at room temperature, and the reaction will continue for 0.5 to 5 hours depending on the specific conjugation.

[0364] 2.5 Reaction termination: After the reaction solution reaches the predetermined conjugation time, add an excess of water-soluble small molecule N-acetyl cysteine solution containing a reducing thiol group (1.63 mg / mL), and continue to slowly stir for 30 minutes.

[0365] 2.6 Product preliminary purification: After the termination reaction of conjugation is completed, add buffer-3 to adjust the pH of the reaction solution to about 5.50; the obtained reaction solution is filtered and subjected to preliminary purification using a G25 dextran gel column, and the component effluent collected in the early stage (about 80%) is concentrated again by ultrafiltration, sterile filtered and subjected to sample dispensing to obtain ADC-01 (wherein b represents DAR). Except for the part of the sample reserved for product analysis, which is stored at 4°C for short-term storage, the other products are stored at -80°C for use.

[0366] The DAR of the prepared ADC-01 based on the quinone structure is 3.23. The determination of the DAR value can be performed according to the method described in the literature (J. Ouyang, in Antibody-Drug Conjugates, Vol. 1045 (Ed.: L. Ducry), 2013, pp. 275-283.).

[0367] Example 3: Preparation of ADC-02 based on quinone structure linker-KRM-1657

[0368] The reaction equation is as follows:

[0369]

[0370] M12 (440 mg, 0.74 mmol) was dissolved in a mixed solution of 4 mL of N,N- dimethylformamide and 5 mL of dichloromethane and cooled at -8°C, and thionyl chloride (170 mg, 1.42 mmol) was slowly added in batches dropwise, and the reaction was carried out at low temperature for 90 minutes. After the reaction was completed, the solvent was removed under reduced pressure to obtain a yellow solid residue. Ethyl ether was added for dispersion, and the slurry was ultrasonically treated, and the filter cake was washed with ethyl ether several times to obtain a light yellow solid powder M14 395 mg, 85.7%.

[0371] M14 (24.96 mg, 0.04 mmol), KRM-1657 (25 mg, 0.027 mmol), N,N- diisopropylethylamine (17.5 mg, 0.135 mmol) were dissolved in 8 mL of N,N- dimethylformamide, stirred at room temperature for 48 h. After the reaction was completed, the solvent was removed under reduced pressure to obtain a blue solid residue, which was further purified by column chromatography to obtain a blue solid powder M15 (31 mg, 75.80% yield). 1 H-NMR (400 MHz, DMSO-d6) δ 9.62 (s, 1H), 9.47 (s, 1H), 8.35 (s, 1H), 8.28 (s, 1H), 7.58 (d, J = 6.0 Hz, 2H), 7.49 (d, J = 6.0 Hz, 2H), 7.16 (t, J = 4.6 Hz, 1H), 6.70 (s, 3H), 6.50-6.48 (m, 1H), 6.45 (d, J = 1.7 Hz, 2H), 6.38 (dd, J = 10.8, 0.8 Hz, 1H), 6.28-6.25 (m, 1H), 6.05-5.98 (m, 1H), 5.14 (dd, J = 4.5, 1.3 Hz, 1H), 5.00-4.95 (m, 1H), 4.72 (s, 2H), 4.41 (d, J = 4.4 Hz, 1H), 3.99-3.97 (m, 4H), 3.94 (d, J = 4.6 Hz, 2H), 3.90-3.88 (m, 4H), 3.83 (d, J = 4.6 Hz, 1H), 3.79 (t, 4.7 Hz, 2H), 3.75-3.70 (m, 1H), 3.66-3.63 (m, 1H), 3.60 (t, J = 3.5 Hz, 2H), 3.37-3.31 (m, 1H), 3.27 (d, J = 1.2 Hz, 3H), 2.82 (s, 2H), 2.61 (s, 3H), 2.41-2.35 (m, 1H), 2.23 (t, J = 6.6 Hz, 2H), 2.16 (d, J = 5.2 Hz, 6H), 2.07 (t, J = 2.0 Hz, 3H), 2.06 (s, 3H), 1.84-1.62 (m, 8H), 1.54-1.48 (m, 2H), 1.43 (s, 5H), 1.41-1.37 (m, 2H), 1.26-1.19 (m, 1H), 1.05-1.01 (m, 7H), 0.92-0.88 (m, 7H), 0.81 (dt, J = 5.8, 1.2 Hz, 3H). HR-MS m / z [M+H] + calculated for C 82 H 99 N8O 20 +: 1515.69, found 1516.69 [M+H] + .

[0372] Referring to Example 2, M15 was conjugated with anti-Golden Staphylococcus antibody to obtain ADC-02 (where b represents DAR). The anti-Golden Staphylococcus antibody was obtained by outsourcing expression, and its sequence is referred to in the literature Novel antibody-antibiotic conjugate eliminates intracellular S. aureus, 2015, Nature, 10.1038 / nature16057. The DAR value of ADC-02 was 3.53.

[0373] Example 4: Preparation of ADC-03 based on benzoquinone structure linker-PBTZ-169

[0374] The equation of the reaction is as follows:

[0375]

[0376] Referring to Example 3, M16 was prepared by replacing KRM-1657 with PBTZ-169. 1 H-NMR (400 MHz, DMSO-d6) δ 9.62 (s, 1H), 8.58 (d, J = 2.6 Hz, 1H), 8.40 (s, 1H), 8.35 (d, J = 2.0 Hz, 1H), 7.62-7.60 (m, 2H), 7.48 (d, J = 6.0 Hz, 2H), 7.16 (t, J = 4.7 Hz, 1H), 6.70 (s, 2H), 5.06 (s, 2H), 4.27-4.25 (m, 4H), 3.94 (d, J = 4.6 Hz, 2H), 3.79 (d, J = 4.67 Hz, 2H), 3.17-3.15 (m, 4H), 2.82 (s, 2H), 2.61 (s, 2H), 2.58 (d, J = 4.1 Hz, 2H), 2.23 (t, J = 6.7 Hz, 2H), 2.15 (s, 3H), 1.72-1.67 (m, 2H), 1.65-1.58 (m, 1H), 1.55-1.47 (m, 8H), 1.43 (s, 6H), 1.41-1.35 (m, 5H), 1.28-1.22 (m, 2H). HR-MS m / z [M+H] + calculated for C 52 H 60 F3N8O 10 S + : 1045.41, found 1046.41 [M+H] + .

[0377] Referring to Example 2, M16 was conjugated with anti-tuberculosis antibody to obtain ADC-03 (wherein b represents DAR). The anti-tuberculosis antibody was obtained by commission expression, and its sequence is referred to WO 2017139153, and the DAR value of ADC-03 is 3.13.

[0378] Example 5: Preparation of ADC-04 based on quinone structure linker-Q203

[0379] The equation of the reaction is as follows:

[0380]

[0381] Referring to Example 3, M17 was prepared by replacing KRM-1657 with Q-203. 1 H-NMR (400 MHz, DMSO-d6) δ 9.62 (s, 1H), 9.18 (d, J = 1.2 Hz, 1H), 8.40 (s, 1H), 8.24 (t, J = 1.6 Hz, 1H), 8.03-7.99 (m, 2H), 7.57-7.52 (m, 5H), 7.46-7.41 (m, 3H), 7.32-7.30 (d, J = 7.0 Hz, 1H), 7.17-7.10 (m, 5H), 6.70 (s, 2H), 5.25 (s, 2H), 4.57 (d, J = 4.2 Hz, 2H), 4.40-4.35 (m, 2H), 4.27-4.22 (m, 2H), 3.94 (d, J = 4.6 Hz, 2H), 3.79 (t, J = 5.0 Hz, 2H), 3.14-3.10 (m, 1H), 2.90 (q, J = 5.6 Hz, 2H), 2.82 (s, 2H), 2.61 (s, 3H), 2.47-2.41 (m, 2H), 2.24-2.17 (m, 4H), 2.15 (s, 3H), 1.69 (p, J = 4.7 Hz, 2H), 1.53-1.47 (m, 2H), 1.43 (s, 5H), 1.42-1.37 (m, 2H), 1.28 (t, J = 5.6 Hz, 3H). HR-MS m / z [M+H] + calculated for C 61 H 65 ClF3N8O9 + :1145.45, found 1146.45[M+H] + .

[0382] Referring to Example 2, M17 was conjugated with anti-tuberculosis antibody to obtain ADC-04 (wherein b represents DAR), and its DAR value is 3.35.

[0383] Example 6: Plasma stability of prepared ADC-01 to ADC-04

[0384] ADC-01 to ADC-04 (15 μL, 700 μg / mL) were incubated in mouse plasma (285 μL) at 37 °C for different time. The required working solution was added into 1.1-mL micro-culture tubes (n = 3), the tubes were gently shaken and vortexed to ensure homogeneity, and then they were placed in a water bath at 37 °C. At 0 h, 4 h, 8 h, 12 h, 24 h, 48 h, 96 h and 168 h after the start of the reaction, 30 μL of the reaction mixture was taken and added to 90 μL of reaction termination solution. The mixture was vortexed for about 1 min, and then centrifuged at 4 °C and 3500 rpm for 10 min. Subsequently, 100 μL of the supernatant was taken for subsequent determination.

[0385] According to the established MMAE standard curve, the concentration of MMAE in the samples at different time points was determined using LC-MS / MS, and the plasma stability data of ADC-01 was obtained.

[0386] According to the established KRM-1657 standard curve, the concentration of KRM-1657 in the samples at different time points was determined using LC-MS / MS, and the plasma stability data of ADC-02 was obtained.

[0387] According to the established PBTZ-163 standard curve, the concentration of PBTZ-163 in the samples at different time points was determined using LC-MS / MS, and the plasma stability data of ADC-03 was obtained.

[0388] According to the established Q-203 standard curve, the concentration of Q-203 in the samples at different time points was determined using LC-MS / MS, and the plasma stability data of ADC-04 was obtained.

[0389] The plasma stability data is shown in Table 1, and the results show that ADC-01 to ADC-04 can remain stable in plasma.

[0390] Table 1 Plasma stability of ADC-01 to ADC-04

[0391]

[0392] Example 7: Drug release behavior of prepared ADC-01 to ADC-04 in the presence of NQO1

[0393] ADC-01 to ADC-04 (90 μL, 140 μg / mL), quinone oxidoreductase 1 (NAD(P)H:quinone oxidoreductase 1, NQOl, 90 μL, 120 μg / mL), reduced coenzyme II (NADPH, 90 μL, 360 μM) and PBS (90 μL, pH 7.4) were added into 1.1 mL micro-culture tubes (n=3). The tubes were shaken gently and vortexed to ensure uniform mixing, and then they were placed in a water bath at 37°C. After the reaction started, 30 μL of the reaction solution was taken at 0 min, 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h, 12 h and 24 h, and 90 μL of the reaction termination solution was added thereto. The mixture was vortexed for about 1 min, and then centrifuged at 4°C and 3500 rpm for 10 min. 100 μL of the supernatant was taken for subsequent determination. According to the standard curve described above, the concentration of the released drug in the samples at different time points was determined using LC-MS / MS. The drug release data are shown in Table 2, and the results show that ADC-01 to ADC-05 can release drugs in the presence of NQOl enzyme.

[0394] Table 2 Drug release of ADC-01 to ADC-04 in the presence of NQOl enzyme

[0395]

[0396] Example 8: Anti-tumor activity of ADC-01

[0397] Cell proliferation was detected by Cell Counting Kit-8 (CCK8, Japan Tongren). The cytotoxic effects of ADC-01 and MMAE were detected. U251 (ATCC CRL-3304), T98G (ATCC CRL-1690) cells were seeded at about 8 x 10 3 The cells were seeded into 96-well plates at a density of 8 x 10 50 .

[0398] IC of ADC-01 to U251 cells 50 = 0.03 nM, IC of ADC-01 to T98G cells 50 = 0.03 nM, IC of MMAE to U251 cells 50= 0.01 nM, IC50 = 0.02 nM for T98G cells 50 = 0.02 nM. ADC-01 showed comparable anti-tumor activity as MMAE.

[0399] Example 9: Anti-S. aureus activity of ADC-02

[0400] (1) Cell plating: well-grown RAW264.7 (provided by Nanjing Medical University) or A549 (provided by Nanjing Medical University) cells were plated in 24-well plates at about 2 x 10 5 cells per well and incubated at 37 °C for 24 h.

[0401] (2) Bacteria preparation: S. aureus USA300 (provided by Nanjing Medical University) was scraped from solid plates and grown in LB liquid medium at 37 °C, 200 rpm overnight. The next day, the overnight grown S. aureus USA300 liquid culture was transferred to fresh LB medium at a ratio of 1 : 100, shaken until OD600= 0.6, and centrifuged at 4700 rpm for 10 min to collect the bacteria, which were resuspended in 1640 medium (containing 10% FBS) and incubated at 37 °C for 30 min. The bacteria were then adjusted to OD600= 0.05 and ready for use.

[0402] (3) Bacterial infection: the medium of the 24-h incubated adherent cells was discarded, and the cells were washed twice with PBS solution. The adjusted S. aureus USA300 bacteria solution was added at a ratio of 1 : 100, and the cells were infected for 2 h.

[0403] (4) Killing of extracellular bacteria: after 2 h of bacterial infection, the medium was discarded, and the cells were washed twice with PBS solution. The cells were treated with 100 μg / mL gentamicin for 1 h to kill the bacteria that did not enter the cells.

[0404] (5) Evaluation of the intracellular bacteria killing ability of ADC-02: after 1 h of gentamicin treatment, the medium was discarded, and the cells were washed twice with PBS solution. The tested compound ADC-02 was added at the corresponding concentration, and the cells were incubated at 37 °C for 24 h.

[0405] (6) Bacterial counting: after 24 h, the medium was discarded, and the cells were washed three times with PBS solution. The cells were resuspended with 1 ml of 1% saponin per well, lysed, diluted, and plated at 37 °C for 24 h.

[0406] (7) Result statistics: after 24 h, the number of viable bacterial colonies in each group was counted and expressed as CFU / ml. The number of colonies in the blank group was taken as 100%, and the survival rate of intracellular S. aureus USA300 was calculated.

[0407] The results show that the bacterial load of RAW264.7 cells treated with 10 μg / ml ADC-02 is reduced to 6.775 CFU / mL. When the treatment concentration of ADC-02 is increased to 100 μg / ml, the bacterial load of RAW264.7 cells is reduced by about 10 5 times, almost reaching the detection limit.

[0408] Example 10: Anti-tuberculosis activity of ADC-03 and ADC-04

[0409] (1) Cell plating: J774A.1 cells (purchased from Beijing Union Cell Resource Center) were cultured in 75 cm 2 cell culture dishes with DMEM medium containing 10% fetal bovine serum. When the cells grew to 80% of the area of the culture dish, the cells were collected after trypsin digestion, resuspended with 5 mL of culture medium. The cell concentration was adjusted to about 4 x 10 5 cells / mL, 1 mL of cell suspension was added to each well of a 24-well cell culture plate, and the adherent cells were incubated at 37°C in a 5% CO2 incubator for 16 h for infection with Mycobacterium tuberculosis.

[0410] (2) Bacterial solution preparation: Mycobacterium tuberculosis H37Rv (strain preserved in the Drug Research Laboratory of the Beijing Tuberculosis and Thoracic Tumor Institute) was inoculated in 7H9 liquid medium containing 10% OADC and 0.05% Tween 80, and cultured at 37°C in a 5% CO2 incubator for 3 weeks to the logarithmic growth phase. The bacterial suspension was filtered through an 8 μm filter to obtain a single bacterial suspension, and the absorbance of the single bacterial suspension was measured at 570 nm using a multifunctional enzyme marker. OD570 0.1 corresponds to a bacterial concentration of 10 7 CFU / mL.

[0411] (3) Bacterial infection: The culture medium of the adherent cells cultured in the 24-well plate for 16 h was discarded, and the cells were washed twice with PBS solution. The adjusted H37Rv bacterial solution was added at a multiplicity of infection (MOI) of 5:1, and the cells were infected for 4 h.

[0412] (4) Removal of extracellular bacteria: After incubation at 37°C for 4 h, the culture medium was discarded, and the cells were washed twice with PBS solution to remove extracellular Mycobacterium tuberculosis.

[0413] (5) Evaluation of the ability of the test compound to kill intracellular bacteria: 2 mL of fresh 10% fetal bovine serum DMEM medium containing different concentrations of the compound was added to each well, and duplicate wells and negative control wells were set up. The cells were incubated at 37°C for 72 h. ADC-03 and ADC-04 were each set at three dosing concentrations, i.e., 1 μg / mL, 5 μg / mL, and 10 μg / mL.

[0414] (6) Bacterial counting: 72 h after the drug effect infection, the culture medium was discarded, 200 μL of 0.1% SDS was added to each well, and lysis was performed at 37°C for 5 min. Then 800 μL of fresh culture medium was added to neutralize the lysis, and after mixing, 10-fold dilution was performed with normal saline. 100 μL of different concentrations of diluents were inoculated on 7H10 solid culture plates, and incubated at 37°C in a 5% CO2 incubator for 3 weeks.

[0415] (7) Result statistics: After 3 weeks, the viable bacterial colonies of each group were counted and expressed as CFU / ml.

[0416] The results show that at a dose of 1 μg / ml, ADC-03 and ADC-04 reduce the number of intracellular bacteria by 1.39 and 1.18 log10 CFU / ml, respectively, both of which exceed 1.0 log10 CFU / ml. At a dose of 5 μg / ml, ADC-03 and ADC-04 reduce the number of bacteria by 1.69 and 2.59 log10 CFU / ml, respectively.

[0417] Although the specific embodiments of the present application have been described in detail, those skilled in the art will understand that various modifications and substitutions can be made to those details based on all the teachings disclosed herein, and such changes are within the scope of protection of the present application. The entire scope of the present application is given by the appended claims and any equivalents thereof.

Claims

1. The compound of Formula I or thereof, in racemic, tautomer, stereoisomer, enantiomer, diastereomer, or mixture of the foregoing configurations, or in pharmaceutically acceptable salt, solvate, or hydrate form, wherein... R1 is hydrogen or C. 1-6 alkyl; R2 is hydrogen or C. 1-6 alkyl; X1 is -O-, -S-, or NR7-, where R7 is hydrogen or C. 1-6 alkyl; Z can be -CH2-, -CH(CH3)-, -C(CH3)2-, or -CH2-NH-; R3 is a hydroxyl group, halogen, or... Where R6 is hydrogen, halogen, or C. 1-6 Alkyl, nitro or C 1-6 Alkoxy group, p is 0, 1, 2, 3 or 4; R5 represents hydrogen, halogen, or C. 1-6 Alkyl, nitro or C 1-6 Alkoxy group; r is 0, 1, 2, 3 or 4; X2 is -O-, -S-, or NR8-, where R8 is hydrogen or C. 1-6 alkyl; L stands for -(CH2) m -C(O)NH-(CH2CH2O) n -(CH2)2-C(O)NH-(CH2) t -C(O)-, -(CH2) m -C(O)NH-(CH2CH2O) n -C(O)NH-(CH2) t -C(O)-、-(CH2CH2O) n -(CH2) m -C(O)-, -(CH2) m -(CH2CH2O) n -C(O)-、 Or -(CH2) m -C(O)NH-(CH2) t -C(O)-, where each t is independently 1, 2, 3, or 4, each m is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12, and each n is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; and R4 is 2. The compound of claim 1 or its racemic, tautomer, stereoisomer, enantiomer, diastereomer, or mixture of the foregoing configurations, or its pharmaceutically acceptable salt, solvate, or hydrate, characterized by the following features One or more: 1) R1 is hydrogen or C 1-4 alkyl; 2) R2 is hydrogen or C 1-4 alkyl; 3) X1 is -O- or -NR7-, wherein R7 is defined as described in claim 1; 4) Z is -CH2- or -CH2-NH-; 5) R3 is a hydroxyl group, fluorine, chlorine, bromine, iodine, or... R6 and p are defined as described in claim 1; 6) R5 is hydrogen, fluorine, chlorine, bromine, iodine, or C. 1-4 Alkyl, nitro or C 1-4 Alkoxy; 7) X2 is -O- or -NR8-, wherein R8 is defined as described in claim 1; 8) L is -(CH2) m -C(O)NH-(CH2CH2O) n -(CH2)2-C(O)NH-(CH2) t -C(O)-, -(CH2) m -C(O)NH-(CH2CH2O) n -C(O)NH-(CH2) t -C(O)-、-(CH2CH2O) n -(CH2) m -C(O)-, -(CH2) m -(CH2CH2O) n -C(O)- or -(CH2) m -C(O)NH-(CH2) t -C(O)-, where t, m, and n are defined as described in claim 1; 9) R4 is 3. The compound of claim 1 or 2, or its racemic, tautomer, stereoisomer, enantiomer, diastereomer, or mixture of the foregoing configurations, or its pharmaceutically acceptable salt, solvate, or hydrate, is characterized by the following features One or more: 1) R1 is hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, preferably hydrogen, methyl, ethyl or n-propyl, and more preferably hydrogen, methyl or ethyl; 2) R2 is hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, preferably hydrogen, methyl, ethyl or n-propyl, and more preferably hydrogen, methyl or ethyl; 3) X1 is -NR7-, where R7 is defined as described in claim 1; 4) Z is -CH2-; 5) R3 is chlorine or R6 and p are defined as described in claim 1; 6) R5 is hydrogen, fluorine, chlorine, bromine, iodine, methoxy, ethoxy, propoxy, butoxy, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl or tert-butyl, preferably hydrogen, fluorine, chlorine, bromine, iodine, methoxy, ethoxy, propoxy, methyl, ethyl, n-propyl or isopropyl, more preferably hydrogen, fluorine, chlorine, bromine, methoxy, ethoxy, methyl or ethyl; 7) X2 is -NR8-, where R8 is defined as described in claim 1; 8) L is -(CH2) m -C(O)NH-(CH2CH2O) n -(CH2)2-C(O)NH-(CH2) t -C(O)- or -(CH2) m -C(O)NH-(CH2) t -C(O)-, where t, m, and n are defined as described in claim 1.

4. The compound of any one of claims 1-3, or a racemic, tautomer, stereoisomer, enantiomer, diastereomer, or mixture of the aforementioned configurations, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, characterized by the following features One or more: 1) R7 is hydrogen or C 1-4 Alkyl groups, preferably hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, or tert-butyl, more preferably hydrogen, methyl, ethyl, or n-propyl, and even more preferably hydrogen, methyl, or ethyl. 2) R8 is hydrogen or C 1-4 Alkyl groups, preferably hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, or tert-butyl, more preferably hydrogen, methyl, ethyl, or n-propyl, and even more preferably hydrogen, methyl, or ethyl. 3) r is 0, 1, 2 or 3, preferably 0, 1 or 2, and more preferably 0 or 1; 4) p is 0, 1, 2 or 3, preferably 0, 1 or 2, and more preferably 0 or 1; 5) R6 is hydrogen, fluorine, chlorine, bromine, iodine, methoxy, ethoxy, propoxy, butoxy, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl or tert-butyl, preferably hydrogen, fluorine, chlorine, bromine, iodine, methoxy, ethoxy, propoxy, methyl, ethyl, n-propyl or isopropyl, more preferably hydrogen, fluorine, chlorine, bromine, methoxy, ethoxy, methyl or ethyl; 6) t is 1, 2 or 3, preferably 1 or 2; 7) m is 1, 2, 3, 4, 5, 6, 7 or 8, preferably 2, 3, 4, 5, 6, 7 or 8, more preferably 4, 5 or 6, or more preferably 7 or 8; 8) n is 1, 2, 3, 4, 5, 6, 7 or 8, preferably 2, 3, 4, 5, 6, 7 or 8, more preferably 4, 5 or 6, or more preferably 7 or 8.

5. The compound of any one of claims 1-4, or a racemic, tautomer, stereoisomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein the compound is selected from: Wherein the definitions of m, n and t are as described in any one of claims 1-4; Preferably, the compound is selected from:

6. Use of the compound of any one of claims 1-5 or its racemic, tautomer, stereoisomer, enantiomer, diastereomer, or mixture of the foregoing configurations, or its pharmaceutically acceptable salts, solvates, or hydrates in the preparation of antibody-drug conjugates.

7. A coupling compound or thereof in the form of a racemate, tautomer, stereoisomer, enantiomer, diastereomer, or mixture of the aforementioned configurations, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, obtained by coupling a compound of any one of claims 1-5 or thereof in the form of a racemate, tautomer, stereoisomer, enantiomer, diastereomer, or mixture of the aforementioned configurations, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, with an active compound. Preferably, the coupling reaction is achieved by coupling the N atom in the active compound molecule with the R3 group in the compound of any one of claims 1-5 to form a carbamate coupling or a quaternary ammonium salt coupling. Preferably, the coupling is as shown in Formula II. The definitions of R1, R2, X1, Z, R5, r, X2, L, and R4 are as described in any one of claims 1-5; Y is -OC(O)- or does not exist; B is an active compound; Preferably, B is selected from drugs, cytotoxins, detection reagents, diagnostic reagents, or targeting carriers; preferably, B is a cytotoxin, antitumor drug, antiviral drug, anti-infective drug, or immunomodulatory drug; more preferably, B is a cytotoxin or anti-infective drug, such as a tubulin inhibitor, antituberculosis drug, DNA alkylating agent, DNA chimera, enzyme inhibitor, antimetabolite, peptide or nucleotide, quinolone antibiotic, or rifamycin antibiotic; even more preferably, B is selected from: auristatin, monomethyl auristatin... Statin E (MMAE), maytansine or its derivatives (e.g., maytansine-like drugs, DM1, DM3, DM4), paclitaxel, calcitrinin, pyroxine, doxorubicin, camptothecin, PBD (pyrrolobenzodiazepines) cytotoxic agents and their derivatives, rifamycin and its derivatives, quinolone antibiotics, antituberculosis drug bedaquiline, antituberculosis drug delamani, antituberculosis drug Q-203, antituberculosis drug PBTZ-169, KRM-1657; More preferably, B is monomethylorisstatin E (MMAE), rifamycin and its derivatives, quinolone antibiotics, anti-tuberculosis drug bedaquiline, anti-tuberculosis drug delamani, anti-tuberculosis drug Q-203, anti-tuberculosis drug PBTZ-169, or KRM-1657.

8. The conjugate of claim 7 or its racemic, tautomer, stereoisomer, enantiomer, diastereomer, or mixture of the foregoing configurations, or its pharmaceutically acceptable salt, solvate, or hydrate, wherein the conjugate is selected from: Wherein the definitions of m, n and t are as described in any one of claims 1-5, and the definition of B is as described in claim 7; Preferably, the coupling agent is selected from:

9. Use of the conjugate of claim 7 or 8 or its racemic, tautomer, stereoisomer, enantiomer, diastereomer, or mixture of the foregoing configurations, or its pharmaceutically acceptable salts, solvates, or hydrates in the preparation of antibody-drug conjugates.

10. An antibody-drug conjugate or thereof in the form of a racemic mixture, tautomer, stereoisomer, enantiomer, diastereomer, or mixture of the aforementioned configurations, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, obtained by conjugation of the conjugate of claim 7 or 8 or thereof in the form of a racemic mixture, tautomer, stereoisomer, enantiomer, diastereomer, or mixture of the aforementioned configurations, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, with an antibody or an antigen-binding fragment thereof. Preferably, the antibody-drug conjugate is as shown in Formula III. The definitions of R1, R2, X1, Z, Y, R5, r, X2, L, and B are as described in claim 7 or 8; A is the target compound, selected from proteins, antibodies, peptides, enzymes, and small molecules; b represents DAR, which is an integer or decimal between 1 and 8.

11. The antibody-drug conjugate of claim 10 or thereof, in the form of a racemic mixture, tautomer, stereoisomer, enantiomer, diastereomer, or mixture of the foregoing configurations, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein A is an antibody or an antigen-binding fragment thereof; Preferably, the antibody or its antigen-binding fragment comprises: (a) The following three heavy chain variable region (VH) complementarity-determining regions (CDRs): (i) VH CDR1, having the sequence of CDR1 contained in VH as shown in SEQ ID NO:1, or having a sequence with one or more amino acid substitutions, deletions or additions (e.g., substitutions, deletions or additions of 1 or 2 or 3 amino acids) compared to the sequence of CDR1 contained in VH. (ii) VH CDR2, having the sequence of CDR2 contained in VH as shown in SEQ ID NO:1, or having a sequence with one or more amino acid substitutions, deletions, or additions (e.g., substitutions, deletions, or additions of one or two amino acids) compared to the sequence of CDR2 contained in VH; and (iii) VH CDR3, having the sequence of CDR3 contained in VH as shown in SEQ ID NO:1, or having a sequence with one or more amino acid substitutions, deletions or additions (e.g., substitution, deletion or addition of 1 amino acid) compared to the sequence of CDR3 contained in VH. and / or (b) The following three light chain variable region (VL) CDRs: (iv) VL CDR1, having the sequence of CDR1 contained in VL as shown in SEQ ID NO:2, or having a sequence with one or more amino acid substitutions, deletions or additions (e.g., substitutions, deletions or additions of 1 or 2 amino acids) compared to the sequence of CDR1 contained in VL. (v) VL CDR2, which has the sequence of CDR2 contained in VL as shown in SEQ ID NO:2, or has a sequence with one or more amino acid substitutions, deletions or additions (e.g., substitutions, deletions or additions of 1 or 2 amino acids) compared to the sequence of CDR2 contained in VL. and (vi) VL CDR3, having the sequence of CDR3 contained in VL as shown in SEQ ID NO:2, or having a sequence with one or more amino acid substitutions, deletions, or additions (e.g., substitutions, deletions, or additions of 1 or 2 amino acids) compared to the sequence of CDR3 contained in VL; or The antibody or its antigen-binding fragment comprises: (a) The following three heavy chain variable region (VH) complementarity-determining regions (CDRs): (i) VH CDR1, having the sequence of CDR1 contained in VH as shown in SEQ ID NO:11, or having a sequence with one or more amino acid substitutions, deletions or additions (e.g., substitutions, deletions or additions of 1, 2 or 3 amino acids) compared to the sequence of CDR1 contained in VH. (ii) VH CDR2, having the sequence of CDR2 contained in VH as shown in SEQ ID NO:11, or having a sequence with one or more amino acid substitutions, deletions, or additions (e.g., substitutions, deletions, or additions of one or two amino acids) compared to the sequence of CDR2 contained in VH; and (iii) VH CDR3, having the sequence of CDR3 contained in VH as shown in SEQ ID NO:11, or having a sequence with one or more amino acid substitutions, deletions or additions (e.g., substitution, deletion or addition of 1 amino acid) compared to the sequence of CDR3 contained in VH. and / or (b) The following three light chain variable region (VL) CDRs: (iv) VL CDR1, having the sequence of CDR1 contained in VL as shown in SEQ ID NO:12, or having a sequence with one or more amino acid substitutions, deletions or additions (e.g., substitutions, deletions or additions of 1 or 2 amino acids) compared to the sequence of CDR1 contained in VL. (v) VL CDR2, which has the sequence of CDR2 contained in VL as shown in SEQ ID NO:12, or has a sequence with one or more amino acid substitutions, deletions or additions (e.g., substitutions, deletions or additions of 1 or 2 amino acids) compared to the sequence of CDR2 contained in VL. and (vi) VL CDR3, having the sequence of CDR3 contained in VL as shown in SEQ ID NO:12, or having a sequence with one or more amino acid substitutions, deletions, or additions (e.g., substitutions, deletions, or additions of one or two amino acids) compared to the sequence of CDR3 contained in VL; or The antibody or its antigen-binding fragment comprises: (a) The following three heavy chain variable region (VH) complementarity-determining regions (CDRs): (i) VH CDR1, having the sequence of CDR1 contained in VH as shown in SEQ ID NO:21, or having a sequence with one or more amino acid substitutions, deletions or additions (e.g., substitutions, deletions or additions of 1, 2 or 3 amino acids) compared to the sequence of CDR1 contained in VH. (ii) VH CDR2, having the sequence of CDR2 contained in VH as shown in SEQ ID NO:21, or having a sequence with one or more amino acid substitutions, deletions, or additions (e.g., substitutions, deletions, or additions of one or two amino acids) compared to the sequence of CDR2 contained in VH; and (iii) VH CDR3, having the sequence of CDR3 contained in VH as shown in SEQ ID NO:21, or having a sequence with one or more amino acid substitutions, deletions or additions (e.g., substitution, deletion or addition of 1 amino acid) compared to the sequence of CDR3 contained in VH. and / or (b) The following three light chain variable region (VL) CDRs: (iv) VL CDR1, having the sequence of CDR1 contained in VL as shown in SEQ ID NO:22, or having a sequence with one or more amino acid substitutions, deletions or additions (e.g., substitutions, deletions or additions of 1 or 2 amino acids) compared to the sequence of CDR1 contained in VL. (v) VL CDR2, which has the sequence of CDR2 contained in VL as shown in SEQ ID NO:22, or has a sequence with one or more amino acid substitutions, deletions or additions (e.g., substitutions, deletions or additions of 1 or 2 amino acids) compared to the sequence of CDR2 contained in VL. and (vi) VL CDR3, having the sequence of CDR3 contained in VL as shown in SEQ ID NO:22, or having a sequence with one or more amino acid substitutions, deletions or additions (e.g., substitutions, deletions or additions of 1 or 2 amino acids) compared to the sequence of CDR3 contained in VL. Preferably, the substitution is a conservative substitution; Preferably, the CDR1, CDR2, and CDR3 contained in the heavy chain variable region (VH), and / or the CDR1, CDR2, and CDR3 contained in the light chain variable region (VL) are defined by the Kabat, Chothia, or IMGT numbering system.

12. The antibody-drug conjugate of claim 11 or its racemic, tautomer, stereoisomer, enantiomer, diastereomer, or mixture of the foregoing configurations, or its pharmaceutically acceptable salt, solvate, or hydrate, wherein the antibody or its antigen-binding fragment comprises: The sequences of CDR1, CDR2, and CDR3 contained in VH as shown in SEQ ID NO:1; and / or The CDR1, CDR2, and CDR3 sequences contained in VL as shown in SEQ ID NO:2; or The antibody or its antigen-binding fragment comprises: The sequences of CDR1, CDR2, and CDR3 contained in VH as shown in SEQ ID NO:11; and / or The CDR1, CDR2, and CDR3 sequences contained in VL as shown in SEQ ID NO:12; or The antibody or its antigen-binding fragment comprises: The sequences of CDR1, CDR2, and CDR3 contained in VH as shown in SEQ ID NO:21; and / or The CDR1, CDR2 and CDR3 sequences contained in VL as shown in SEQ ID NO:22; Preferably, the CDR1, CDR2 and CDR3 contained in the heavy chain variable region (VH), and / or the CDR1, CDR2 and CDR3 contained in the light chain variable region (VL) are defined by the Kabat, Chothia or IMGT numbering system; Preferably, the antibody or its antigen-binding fragment comprises: (a) The following three heavy chain variable region (VH) CDRs: (i) VH CDR1, which consists of the sequence shown in SEQ ID NO:

3. (ii) VH CDR2, which consists of the sequence shown in SEQ ID NO:4, and (iii) VH CDR3, which consists of the sequence shown in SEQ ID NO:5; and / or (b) The following three light chain variable region (VL) CDRs: (iv) VL CDR1, which consists of the sequence shown in SEQ ID NO:

6. (v)VL CDR2, which consists of the sequence shown in SEQ ID NO:7, and (vi)VL CDR3, which consists of the sequence shown in SEQ ID NO:8; or The antibody or its antigen-binding fragment comprises: (a) The following three heavy chain variable region (VH) CDRs: (i) VH CDR1, which consists of the sequence shown in SEQ ID NO:

13. (ii) VH CDR2, which consists of the sequence shown in SEQ ID NO:14, and (iii) VH CDR3, which consists of the sequence shown in SEQ ID NO:15; and / or (b) The following three light chain variable region (VL) CDRs: (iv) VL CDR1, which consists of the sequence shown in SEQ ID NO:

16. (v)VL CDR2, which consists of the sequence shown in SEQ ID NO:17, and (vi)VL CDR3, which consists of the sequence shown in SEQ ID NO:18; or The antibody or its antigen-binding fragment comprises: (a) The following three heavy chain variable region (VH) CDRs: (i) VH CDR1, which consists of the sequence shown in SEQ ID NO:

23. (ii) VH CDR2, which consists of the sequence shown in SEQ ID NO:24, and (iii) VH CDR3, which consists of the sequence shown in SEQ ID NO:25; and / or (b) The following three light chain variable region (VL) CDRs: (iv) VL CDR1, which consists of the sequence shown in SEQ ID NO:

26. (v)VL CDR2, which consists of the sequence shown in SEQ ID NO:27, and (vi)VL CDR3, which consists of the sequence shown in SEQ ID NO:28; Preferably, the VH of the antibody or its antigen-binding fragment comprises: VH CDR1 as shown in SEQ ID NO:3; VH CDR2 as shown in SEQ ID NO:4; and VH CDR3 as shown in SEQ ID NO:5; and / or, the VL of the antibody or its antigen-binding fragment comprises: VL CDR1 as shown in SEQ ID NO:6; VL CDR2 as shown in SEQ ID NO:7; and VL CDR3 as shown in SEQ ID NO:8; or The VH of the antibody or its antigen-binding fragment comprises: VH CDR1 as shown in SEQ ID NO:13; VH CDR2 as shown in SEQ ID NO:14; and VH CDR3 as shown in SEQ ID NO:15; and / or, the VL of the antibody or its antigen-binding fragment comprises: VL CDR1 as shown in SEQ ID NO:16; VL CDR2 as shown in SEQ ID NO:17; and VL CDR3 as shown in SEQ ID NO:18; or The VH of the antibody or its antigen-binding fragment comprises: VH CDR1 as shown in SEQ ID NO:23; VH CDR2 as shown in SEQ ID NO:24; and VH CDR3 as shown in SEQ ID NO:25; and / or, the VL of the antibody or its antigen-binding fragment comprises: VL CDR1 as shown in SEQ ID NO:26; VL CDR2 as shown in SEQ ID NO:27; and VL CDR3 as shown in SEQ ID NO:28; Preferably, the VH of the antibody or its antigen-binding fragment comprises: VH CDR1 as shown in SEQ ID NO:3; VH CDR2 as shown in SEQ ID NO:4; and VH CDR3 as shown in SEQ ID NO:5; and the VL of the antibody or its antigen-binding fragment comprises: VL CDR1 as shown in SEQ ID NO:6; VL CDR2 as shown in SEQ ID NO:7; and VL CDR3 as shown in SEQ ID NO:8; or The VH of the antibody or its antigen-binding fragment comprises: VH CDR1 as shown in SEQ ID NO:13; VH CDR2 as shown in SEQ ID NO:14; and VH CDR3 as shown in SEQ ID NO:15; and the VL of the antibody or its antigen-binding fragment comprises: VL CDR1 as shown in SEQ ID NO:16; VL CDR2 as shown in SEQ ID NO:17; and VL CDR3 as shown in SEQ ID NO:18; or The VH of the antibody or its antigen-binding fragment comprises: VH CDR1 as shown in SEQ ID NO:23; VH CDR2 as shown in SEQ ID NO:24; and VH CDR3 as shown in SEQ ID NO:25; and the VL of the antibody or its antigen-binding fragment comprises: VL CDR1 as shown in SEQ ID NO:26; VL CDR2 as shown in SEQ ID NO:27; and VL CDR3 as shown in SEQ ID NO:28; Preferably, the antibody or its antigen-binding fragment comprises: (a) Heavy chain variable region (VH), which contains an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 1; (ii) A sequence having one or more amino acid substitutions, deletions, or additions (e.g., substitutions, deletions, or additions of 1, 2, 3, 4, or 5 amino acids) compared to the sequence shown in SEQ ID NO: 1; and (iii) A sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence shown in SEQ ID NO: 1; and (b) Light chain variable region (VL), which contains an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 2; (v) A sequence having one or more amino acid substitutions, deletions, or additions (e.g., substitutions, deletions, or additions of 1, 2, 3, 4, or 5 amino acids) compared to the sequence shown in SEQ ID NO: 2; and (vi) A sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence shown in SEQ ID NO: 2; or The antibody or its antigen-binding fragment comprises: (a) Heavy chain variable region (VH), which contains an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 11; (ii) A sequence having one or more amino acid substitutions, deletions, or additions (e.g., substitutions, deletions, or additions of 1, 2, 3, 4, or 5 amino acids) compared to the sequence shown in SEQ ID NO: 11; and (iii) A sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence shown in SEQ ID NO: 11; and (b) Light chain variable region (VL), which contains an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 12; (v) A sequence having one or more amino acid substitutions, deletions, or additions (e.g., substitutions, deletions, or additions of 1, 2, 3, 4, or 5 amino acids) compared to the sequence shown in SEQ ID NO: 12; and (vi) A sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence shown in SEQ ID NO: 12; or The antibody or its antigen-binding fragment comprises: (a) Heavy chain variable region (VH), which contains an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 21; (ii) A sequence having one or more amino acid substitutions, deletions, or additions (e.g., substitutions, deletions, or additions of 1, 2, 3, 4, or 5 amino acids) compared to the sequence shown in SEQ ID NO: 21; and (iii) A sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence shown in SEQ ID NO: 21; and (b) Light chain variable region (VL), which contains an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 22; (v) A sequence having one or more amino acid substitutions, deletions, or additions (e.g., substitutions, deletions, or additions of 1, 2, 3, 4, or 5 amino acids) compared to the sequence shown in SEQ ID NO: 22; and (vi) A sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence shown in SEQ ID NO: 22; Preferably, the substitution is a conservative substitution; Preferably, the antibody or its antigen-binding fragment comprises: The heavy chain variable region (VH) comprises a sequence having the sequence shown in SEQ ID NO:1 or having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence shown in SEQ ID NO:1; and the light chain variable region (VL) comprises a sequence having the sequence shown in SEQ ID NO:2 or having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence shown in SEQ ID NO:2; or The heavy chain variable region (VH) comprises a sequence having the sequence shown in SEQ ID NO:11 or having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence shown in SEQ ID NO:1; and the light chain variable region (VL) comprises a sequence having the sequence shown in SEQ ID NO:12 or having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence shown in SEQ ID NO:12; or The heavy chain variable region (VH) comprises a sequence having the sequence shown in SEQ ID NO:21 or having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence shown in SEQ ID NO:21; and the light chain variable region (VL) comprises a sequence having the sequence shown in SEQ ID NO:22 or having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence shown in SEQ ID NO:22; Preferably, the amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity does not involve changes to the CDR sequence; Preferably, the antibody or its antigen-binding fragment comprises: VH having the sequence shown in SEQ ID NO:1 and VL having the sequence shown in SEQ ID NO:2; or The antibody or its antigen-binding fragment comprises: VH having the sequence shown in SEQ ID NO:11 and VL having the sequence shown in SEQ ID NO:12; or The antibody or its antigen-binding fragment comprises: VH having the sequence shown in SEQ ID NO:21 and VL having the sequence shown in SEQ ID NO:

22.

13. The antibody-drug conjugate of claim 11 or 12 or its racemic, tautomer, stereoisomer, enantiomer, diastereomer, or mixture of the foregoing configurations, or its pharmaceutically acceptable salt or solvate or hydrate, wherein the antibody or its antigen-binding fragment is humanized. Preferably, the antibody or its antigen-binding fragment further comprises the framework region of a human immunoglobulin; Preferably, the antibody or its antigen-binding fragment further comprises a heavy chain framework region of human immunoglobulin (e.g., a heavy chain framework region contained in the amino acid sequence encoded by the human heavy chain germline antibody gene), and / or a light chain framework region of human immunoglobulin (e.g., a light chain framework region contained in the amino acid sequence encoded by the human light chain germline antibody gene). Preferably, the heavy chain framework region and / or the light chain framework region optionally include one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) reversion mutations from human residues to mouse residues; Preferably, the antibody or its antigen-binding fragment comprises: A heavy chain having the sequence shown in SEQ ID NO:9 and a light chain having the sequence shown in SEQ ID NO:10; or A heavy chain having the sequence shown in SEQ ID NO:19 and a light chain having the sequence shown in SEQ ID NO:20; or A heavy chain having a sequence as shown in SEQ ID NO:29 and a light chain having a sequence as shown in SEQ ID NO:30; Preferably, the antibody or its antigen-binding fragment further comprises a constant region derived from human immunoglobulin; Preferably, the heavy chain of the antibody or its antigen-binding fragment includes a heavy chain constant region derived from human immunoglobulins (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, IgM, IgD, or IgE). Preferably, the light chain of the antibody or its antigen-binding fragment includes a light chain constant region derived from human immunoglobulins (e.g., κ or λ); Preferably, the antibody or its antigen-binding fragment is selected from: (1) Mouse-derived antibodies, chimeric antibodies (e.g., human-mouse chimeric antibodies), humanized antibodies, and fully human antibodies; (2) Full-length antibody, single-chain antibody, Fab, Fab', F(ab')2, Fv, scFv, sdAb, dAb, complementarity-determining region fragment, Fd; (3) Monoclonal antibodies, polyclonal antibodies; and (4) Monospecific antibodies, bispecific antibodies, and multispecific antibodies; Preferably, the monoclonal antibody includes a non-CDR region, and the non-CDR region is derived from a species other than rodents, such as from human antibodies; Preferably, the antibody or its antigen-binding fragment is a full-length antibody; Preferably, the antibody or its antigen-binding fragment is trastuzumab (HERCEPTIN), cetuximab (ERBITUX), bevacizumab (AVASTIN), anti-tuberculosis antibody, anti-Staphylococcus aureus antibody, or its antigen-binding fragment; Preferably, A is coupled to site # by targeting the S or N atom in the compound molecule.

14. The antibody-drug conjugate of any one of claims 11-13, or a racemic, tautomer, stereoisomer, enantiomer, diastereomer, or mixture of the foregoing configurations, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein b is an integer or decimal between 2 and 7. Preferably, b is an integer or decimal between 3 and 6, or an integer or decimal between 4 and 5; Preferably, b is an integer or decimal between 3 and 4; Preferably, b is about 2, about 3, about 4, about 5, or about 6; Preferably, b is an integer or decimal between 3 and 3.6; Preferably, b is about 3.1, about 3.2, about 3.3, about 3.4 or about 3.

5.

15. The antibody-drug conjugate of any one of claims 11-14, or a racemic, tautomer, stereoisomer, enantiomer, diastereomer, or mixture of the foregoing configurations, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein the antibody-drug conjugate is selected from: The definitions of A, m, n, t, B, and b are as described in any one of claims 10-14; Preferably, the antibody-drug conjugate is selected from: Wherein the definitions of A, B, and b are as described in any one of claims 10-14; preferably, the antibody-drug conjugate is selected from: The definitions of A and b are as described in any one of claims 10-14.

16. A pharmaceutical composition comprising at least one antibody-drug conjugate of any one of claims 10-15, or a racemic, tautomer, stereoisomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, and one or more pharmaceutically acceptable carriers or excipients.

17. Use of any one of the antibody-drug conjugates of claims 10-15, or in the form of a racemic, tautomer, stereoisomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, in the preparation of a medicament, wherein the medicament is intended to treat a disease or condition or to reduce the severity of said disease or condition in a subject, said disease or condition being selected from tumors, bacterial infections, infectious diseases, hematological diseases, metabolic diseases, and inflammation; Preferably, the tumor is selected from cancer, lymphoma, lymphoid tumor, blastoma, sarcoma, leukemia, and glioma; Preferably, the infectious disease is caused by intracellular bacterial infection; Preferably, the infectious disease is caused by Mycobacterium tuberculosis or Staphylococcus aureus infection. Preferably, the bacterial infection is Mycobacterium tuberculosis infection or Staphylococcus aureus infection; Preferably, the cancer is selected from: breast cancer (e.g., HER2-positive breast cancer); squamous cell carcinoma (e.g., epithelial squamous cell carcinoma); lung cancer, including small cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, and squamous cell carcinoma of the lung; peritoneal cancer; liver cancer; gastric cancer; gastrointestinal cancer; mesenteric adenocarcinoma; glioblastoma; cervical cancer; ovarian cancer; bladder cancer; urethral cancer; hepatocellular carcinoma; intestinal cancer; colon cancer; rectal cancer; colorectal cancer; endometrial cancer; uterine cancer; salivary gland cancer; kidney cancer; prostate cancer; vulvar cancer; thyroid cancer; anal cancer; penile cancer; melanoma; multiple myeloma and B-cell lymphoma; brain cancer; gallbladder cancer; esophageal cancer; bile duct cancer; head and neck cancer and related metastases.

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