Linker-drug molecule and antibody drug conjugate, preparation method therefor and use thereof

Abstract

A compound represented by formula (1), or a tautomer, mesomer, racemate, enantiomer, and diastereomer thereof, or mixture form thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the structure of formula (1) is as shown below: formula (1). According to the compound, a strong hydrophilic sulfonate inner salt and / or phosphate inner salt side chain are introduced at the position of polypeptide (VC, VA, AAA, etc.)-PAB (self-immolative group) of a linker, greatly enhancing the druggability (comprising hydrophilicity and stability) of an antibody drug conjugate corresponding to the compound, reducing the in-vivo drug clearance effect, and improving the tumor treatment effect.

Description

A linker-drug molecule and antibody-drug conjugate, preparation method and application thereof Technical Field

[0001] The present application relates to the field of biomedicine, and specifically to a linker-drug molecule and an antibody-drug conjugate, and their preparation methods and applications.

[0002] Cross-references

[0003] This application claims the benefit of PCT International Application No. PCT / CN2023 / 106385, filed on July 7, 2023, the contents of which are incorporated herein in their entirety.

[0004] Sequence listing file submitted at the same time

[0005] The entire contents of the following XML file are incorporated herein by reference in their entirety: Sequence Listing in Computer Readable Format (CRF) (Name: TFH01015PCT-SequenceListing.xml, Date: 2024.7.2, Size: 6KB). Background Art

[0006] By conjugating biologically active drug compounds to tumor-targeting antibodies, proteins, or peptides, the cancer therapeutic window of the drug compound can be increased. This is achieved primarily through two factors: first, the drug conjugate cannot produce toxic side effects before reaching the cancer target lesion because the active drug cannot be smoothly detached; second, the active drug can increase the concentration of the drug compound in the cancer target lesion through the conjugated targeting antibody, protein, or peptide, while the specially designed linker can be stimulated by the unique environment of the cancer target lesion to break and release the active drug compound.

[0007] The specially designed cathepsin-sensitive amino acid peptide in the linker is efficiently cleaved by cathepsins in the lysosomes of cancer cells. The drug compound is attached to the amino acid peptide of the linker via a chemical self-immolative group. After the cathepsin cleaves the peptide, the chemical self-immolative group releases the free drug compound through a "self-immolative effect," thereby producing a tumor-killing effect. Among these, antibody- or peptide-drug conjugates with linkers in the form of peptides (VC, VA, AAA, etc.)-PAB (self-immolative groups) have the most clinical validation data and approved drugs. However, because drug compounds, especially those with "bystander effects" that are important for tumor killing, have strong lipophilic characteristics, antibody-, protein-, or peptide-drug conjugates, especially at high drug-to-drug ratios (DARs), have poor drugability, manifesting as low hydrophilicity, forming aggregates, and accelerated clearance from the body. By adding hydrophilic groups to the linker, such as polyethylene glycol (PEG) (Mol Cancer Ther, 2017, 16 (1): 116-123; Nat. Biotech. 2015, 33: 733–735, Toxicol. App. Pharmacol., 2020, 392: 114932; WO2023280227) polysarcosine (PSAR) (Chem. Sci, 2019, 10 (14): 4048-4053; WO 2022228493) sugar alcohol (Cancer Res (2018) 78 (13_Supplement): 755; US 9907854), etc., to shield the drug compound, the drugability of the antibody, protein or peptide drug conjugate can be improved to a certain extent. However, the use of PEG polymers to modify proteins has exposed many shortcomings in clinical practice, such as a high proportion of patients with anti-PEG immunogenicity and related toxic side effects caused by it, and excessive modification leading to reduced protein function.

[0008] Summary of the Invention

[0009] Based on this, the present application introduces a strongly hydrophilic sulfonic acid inner salt and / or phosphate inner salt side chain (hereinafter sometimes also referred to as the linker or inner salt linker of the present application, i.e., the compound represented by formula (1) described below) at the polypeptide (VC, VA, AAA, etc.)-PAB (self-destructive group) position of the linker. Since the sulfonic acid group or the phosphate group has both a hydrogen bond donor and a hydrogen bond acceptor, it is a strongly hydrophilic group, and the sulfonic acid inner salt or the phosphate inner salt has both a positive charge and a negative charge, which can further increase the hydrophilicity. At the same time, the water-soluble group in the form of the inner salt also has the potential to enhance the tumor targeting of the antibody drug conjugate and reduce nonspecific binding, thereby greatly enhancing the drugability (including hydrophilicity and stability) of the test antibody drug conjugate, reducing the drug clearance effect in the body, and improving the tumor treatment effect.

[0010] The technical solution of this application is as follows:

[0011] 1. A compound represented by formula (1), or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein the structure of formula (1) is as follows:

[0012] M is a linking group; Sp is -L a -Q a -; A is selected from the group consisting of absence, hydrogen, and alkyl; T represents a linear or branched (C1-C8) trivalent alkyl group; Y, Y', and Y" are respectively -L b -Q b -;L a , L b are independently selected from the group consisting of absence, alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene, heteroarylene or polyethylene glycol group, wherein the alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene, heteroarylene or polyethylene glycol group is optionally substituted with a substituent;

[0013] Q a , Q b are each selected from absent, -C(=O)NR-, -C(=O)O-, -OC(=O)NR-, -S(=O)2NR-, -NRC(=O)-, -OC(=O)-, -NRC(=O)O-, -NRS(=O)2-, -NRC(=O)NR-, -NHC-(=NH)NH-, -C(=O)-, -OC(=O)O-, -O-, -NR-, -S-, S(=O)2-, S(=O)-, or -Se-, wherein R is selected from hydrogen or optionally substituted alkyl;

[0014] Z is selected from absent, hydrogen, optionally substituted alkyl, or sulfonate;

[0015] Z', Z" are selected from absent, hydrogen, optionally substituted alkyl, sulfonic acid inner salt or phosphate inner salt;

[0016] At least one of Z, Z', and Z" is a sulfonic acid inner salt or a phosphate inner salt;

[0017] X comprises a self-destruct unit or does not exist; D comprises a drug molecule; i is an integer greater than or equal to 1; h is an integer from 1 to 6; o, p, and q are integers from 0 to 5, respectively.

[0018] 2. The compound according to item 1, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the formula (1) is formula (1-1):

[0019] Wherein, M, Sp, A, T, Y, h, o, i, X, D, and Z are as defined in item 1;

[0020] At least one of the i Zs is a sulfonate.

[0021] 3. The compound according to item 2, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the formula (1-1) is formula (1-1-1):

[0022] Wherein, AA is an amino acid residue, and W is an integer greater than or equal to 1.

[0023] 4. The compound according to item 1, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the formula (1) is formula (1-2):

[0024] Wherein, M, Sp, A, T, Y, Z, o, p, i, h, X, Y′, and D are as defined in item 1;

[0025] Z' is a sulfonic acid inner salt or a phosphoric acid inner salt.

[0026] 5. The compound according to item 4, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the formula (1) is formula (1-2-1):

[0027] Wherein, AA is an amino acid residue, and W is an integer greater than or equal to 1.

[0028] 6. The compound according to item 1, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the formula (1) is formula (1-3):

[0029] wherein M, Sp, A, Y, T, Z, o, i, h, q, X, Y″, and D are as defined in item 1;

[0030] Z″ is a sulfonic acid inner salt or a phosphoric acid inner salt.

[0031] 7. The compound according to item 6, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the formula (1) is formula (1-3-1):

[0032] Wherein, AA is an amino acid residue, and W is an integer greater than or equal to 1.

[0033] 8. The compound according to any one of items 1 to 7, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein when A and Z are absent, N, Y, and T form a ring, namely

[0034] Preferably

[0035] 9. A compound according to any one of items 3, 5 or 7, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the amino acid is selected from one or more of alanine, arginine, asparagine, aspartic acid, γ-carboxyglutamate, citrulline, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, norleucine, norvaline, ornithine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine.

[0036] 10. A compound according to any one of items 3, 5 or 7, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the amino acid is selected from one or more of valine, glutamic acid, citrulline, lysine, alanine, phenylalanine, arginine, glutamine, asparagine and glycine.

[0037] 11. A compound according to any one of items 3, 5 or 7, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the amino acid is selected from one or more of valine, citrulline, alanine, phenylalanine, glutamine and glycine.

[0038] 12. The compound according to any one of items 1 to 11, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein:

[0039] M is selected from N3, SH, ONH2, NH2, CO2H, (alkyl)-CO-, HCO-, BrCH 2- 、ICH2-、

[0040] 13. The compound according to any one of items 1 to 12, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein:

[0041] M is

[0042] 14. The compound according to any one of items 1 to 13, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein:

[0043] M is

[0044] 15. A compound according to any one of items 1 to 14, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein A is hydrogen, methyl, ethyl or propyl.

[0045] 16. The compound according to any one of items 1 to 15, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein A is hydrogen.

[0046] 17. The compound according to any one of items 1 to 16, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein T is one of the following structures:

[0047] 18. The compound according to any one of items 1 to 17, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein T is

[0048] 19. The compound according to any one of items 1 to 18, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein L a One, two or more selected from alkylene, alkenylene, alkynylene, saturated or unsaturated alicyclic group having 3-10 ring carbon atoms, saturated or unsaturated heteroalicyclic group having 3-10 ring carbon atoms and 1 to 4 heteroatoms, arylene group having 6 to 18 carbon atoms in one or more rings, or heteroarylene group having 6 to 18 carbon atoms in one or more rings and 1 to 4 heteroatoms, or polyethylene glycol group, wherein the alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene group and heteroarylene group are optionally substituted by a substituent, and the heteroatom is selected from nitrogen, oxygen or sulfur.

[0049] 20. The compound according to any one of items 1 to 19, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein L a Selected from C1-C 10 Straight or branched alkylene, C2-C 12 Straight or branched alkenylene, C2-C 12 One, two or more of a straight or branched alkynylene group, a saturated or unsaturated alicyclic group having 3-6 ring carbon atoms, a saturated or unsaturated heteroalicyclic group having 3-6 ring carbon atoms and 1 to 4 heteroatoms, or an arylene group having 6 to 10 carbon atoms in one or more rings, a heteroarylene group having 6 to 10 carbon atoms in one or more rings and 1 to 4 heteroatoms, or a polyethylene glycol group; the alkylene group, alkenylene group, alkynylene group, alicyclic group, heteroalicyclic group, arylene group and heteroarylene group are optionally substituted by a substituent, and the heteroatom is selected from nitrogen, oxygen or sulfur.

[0050] 21. The compound according to any one of items 1 to 20, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein L a One, two or more selected from C1-C6 straight chain or branched chain alkylene, C2-C6 straight chain or branched chain alkenylene, C2-C6 straight chain or branched chain alkynylene, or polyethylene glycol group, wherein the alkylene, alkenylene and alkynylene groups are optionally substituted by a substituent.

[0051] 22. The compound according to any one of items 1 to 21, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein L a is one, two or more of a C1-C6 straight or branched alkylene group or a polyethylene glycol group,

[0052] The polyethylene glycol group is -(CH2CH2O) n -, -O(CH2CH2O) n -、-(CH2CH2O) n CH2-, -(CH2CH2O) n CH2CH2-, -CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n CH2-, n is an integer greater than or equal to 1.

[0053] 23. The compound according to any one of items 1 to 22, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein L a is ethylene, linear propylene, linear butylene, linear pentylene, or -(CH2CH2O) n One, two or more of CH2CH2-, wherein n is 2, 3, 4, 5, 6, 7 or 8.

[0054] 24. The compound according to any one of items 1 to 23, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein L a It is ethylene and / or -(CH2CH2O)4CH2CH2- or pentylene.

[0055] 25. The compound according to any one of items 1 to 24, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein L bOne, two or more than three selected from alkylene, alkenylene, alkynylene, saturated or unsaturated alicyclic group having 3-10 ring carbon atoms, saturated or unsaturated heteroalicyclic group having 3-10 ring carbon atoms and 1 to 4 heteroatoms, arylene group having 6 to 18 carbon atoms in one or more rings, or heteroarylene group having 6 to 18 carbon atoms in one or more rings and 1 to 4 heteroatoms, or polyethylene glycol, wherein the alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene group and heteroarylene group are optionally substituted by a substituent, and the heteroatom is selected from nitrogen, oxygen or sulfur.

[0056] 26. The compound according to any one of items 1 to 25, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein L b Selected from C1-C 10 Straight or branched alkylene, C2-C 12 Straight or branched alkenylene, C2-C 12 One or two or more of the group consisting of a straight or branched alkynylene group, a saturated or unsaturated alicyclic group having 3-6 ring carbon atoms, a saturated or unsaturated heteroalicyclic group containing 3-6 ring carbon atoms and 1 to 4 heteroatoms, or an arylene group having 6 to 10 carbon atoms in one or more rings, a heteroarylene group having 6 to 10 carbon atoms in one or more rings and 1 to 4 heteroatoms, or polyethylene glycol, wherein the alkylene group, alkenylene group, alkynylene group, alicyclic group, heteroalicyclic group, arylene group, and heteroarylene group are optionally substituted by a substituent, and the heteroatom is selected from nitrogen, oxygen, or sulfur.

[0057] 27. The compound according to any one of items 1 to 26, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein L b is selected from C1-C6 straight chain or branched chain alkylene, C2-C6 straight chain or branched chain alkenylene, or C2-C6 straight chain or branched chain alkynylene, or polyethylene glycol group,

[0058] The polyethylene glycol group is -(CH2CH2O) n -, -O(CH2CH2O) n -、-(CH2CH2O) n CH2-, -(CH2CH2O) n CH2CH2-, -CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) nOne, two, or more than three of CH2-, n is an integer greater than or equal to 1, and the alkylene, alkenylene, and alkynylene groups are optionally substituted with a substituent.

[0059] 28. The compound according to any one of items 1 to 27, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein L b is a C1-C6 straight or branched chain alkylene, or a C6-C 10 One or two or more than three arylene groups.

[0060] 29. The compound according to any one of items 1 to 28, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein L b It is one or two or more of the following: methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, sec-butylene, tert-butylene, or benzylene.

[0061] 30. The compound according to any one of items 1 to 29, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein Q a One, two or more selected from -C(=O)NR-, -NR-, -C(=O)O-, -OC(=O)NR-, -NRC(=O)-, -OC(=O)-, -NRC(=O)O-, -NRC(=O)NR-, NHC-(=NH)NH-, -C(=O)-, -OC(=O)O-, wherein R is selected from hydrogen, optionally substituted C1-C6 straight chain or branched alkyl.

[0062] 31. The compound according to item 30, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein Q a It is one, two, or more than three of -C(=O)-, -NR-, -C(=O)NR-, -NRC(=O)NR-, -OC(=O)O-, -OC(=O)NR-, -NRC(=O)-, or -OC(=O)-, wherein R is selected from hydrogen or an optionally substituted C1-C6 straight or branched alkyl group.

[0063] 32. The compound according to item 30, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein Q a It is one or two or more of -C(=O)-, -NR-, -NRC(=O)- and -(C=O)NR-, wherein R is selected from hydrogen or an optionally substituted C1-C6 linear or branched alkyl group.

[0064] 33. The compound according to item 30, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein Q a It is -C(=O)-, -N(CH3)-, -NHC(=O)-, -C(=O)-NH- or -C(=O)-N(CH3)-.

[0065] 34. The compound according to any one of items 1 to 33, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein Q b is selected from one, two or more of the group consisting of absent, -C(=O)NR-, -C(=O)O-, -OC(=O)NR-, -NRC(=O)-, -OC(=O)-, -NRC(=O)O-, -NRC(=O)NR-, NHC-(=NH)NH-, -C(=O)-, and -OC(=O)O-, wherein R is selected from hydrogen, an optionally substituted C1-C6 straight chain or branched alkyl group.

[0066] 35. The compound according to item 34, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein Q b is absent, one or two or more of -C(=O)-, -C(=O)NR-, -OC(=O)NR-, -NRC(=O)-, -NRC(=O)O-, or -NRC(=O)NR-, and R is selected from hydrogen, an optionally substituted C1-C6 linear or branched alkyl group.

[0067] 36. The compound according to item 34, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein Q b is absent, one, two, or more than one of -C(=O)-, -NRC(=O)-, -NRC(=O)NR-, or -C(=O)NR-, and R is hydrogen or methyl.

[0068] 37. The compound according to any one of items 1 to 36, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein X is selected from:

[0069] The right wavy line in X connects D; R4 is selected from hydrogen, C1-C6 straight or branched alkyl, -(CH2CH2O) m R2, -CH2CH2NR2R3, -CH2CH2SO2R2, R2, R3 are independently selected from hydrogen, C1-C6 straight chain or branched alkyl, and m is an integer greater than 1.

[0070] 38. The compound according to item 37, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein:

[0071] X is

[0072] R4 is hydrogen, methyl, -CH2CH2OCH2CH2OH, -CH2CH2NMe2, -CH2CH2SO2Me.

[0073] 39. The compound according to item 37, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein:

[0074] X is R4 is hydrogen or methyl.

[0075] 40. The compound according to any one of items 1 to 39, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein the sulfonic acid inner salt is

[0076] A wavy line connects Y, Y' or Y", wherein c, d, e, f and g are integers of 1, 2, 3, 4, 5 or 6;

[0077] R6, R7, R 11 、R 12 、R 13 、R 14 are independently selected from absence, hydrogen, and C1-C6 linear or branched alkyl.

[0078] 41. The compound according to item 40, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein the sulfonic acid inner salt is

[0079] in,

[0080] R6, R7, R 11 、R 12 、R 13 and R 14 are selected from absence, methyl or ethyl, c, d, e, f, g are 1, 2 or 3 respectively.

[0081] 42. The compound according to item 40, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the sulfonic acid inner salt is one of the following structures:

[0082] 43. The compound according to any one of items 1 to 42, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein the phosphate inner salt is

[0083] in,

[0084] a and b are both integers of 1, 2, 3, 4, 5 or 6; R8, R9 and R 10 are independently selected from hydrogen, C1-C6 straight chain or branched alkyl.

[0085] 44. The compound according to item 43, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein:

[0086] The phosphate inner salt is

[0087] Among them, R8, R9, R 10 are methyl or ethyl respectively; a is 1 or 2 or 3; b is 1 or 2 or 3.

[0088] 45. The compound according to item 43, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein:

[0089] The phosphate inner salt is

[0090] Among them, R8, R9, R 10 All are methyl; a is 2; b is 1.

[0091] 46. ​​A compound according to any one of items 1 to 45, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein D comprises a cytotoxin or an immunostimulant.

[0092] 47. A compound according to claim 46, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the cytotoxin is selected from one or two of a microtubule inhibitor, a DNA synthesis inhibitor, a topoisomerase inhibitor or an RNA polymerase 2 inhibitor.

[0093] 48. The compound according to item 46, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein:

[0094] The cytotoxin is selected from auristatins, maytansine alkaloids DMs, calicheamicin, duocarmycin, pyrrolobenzodiazepines One or more of camptothecin derivatives, camptothecin derivatives and α-amanitin.

[0095] 49. The compound according to item 46, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein:

[0096] The immune agonist is selected from TLR7 / 8 agonist or STING agonist.

[0097] 50. A compound according to any one of items 1-49, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein i is 2, 3, 4, 5, 6, 7 or 8.

[0098] 51. A compound according to any one of items 1 to 50, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein h is 1, 2, 3 or 4.

[0099] 52. A compound according to any one of items 1 to 51, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein o, p, q are 0, 1, 2 or 3, respectively.

[0100] 53. The compound according to any one of items 1 to 52, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein L a It is one, two or more of C1-C6 straight or branched alkylene groups, or polyethylene glycol groups, wherein the polyethylene glycol group is -(CH2CH2O) n -, -O(CH2CH2O) n -、-(CH2CH2O) n CH2-, -(CH2CH2O) n CH2CH2-, -CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n CH2-, n is an integer greater than or equal to 1; Q a It is one, two, or more than three of -C(=O)-, -NR-, -NR(C=O)-, and -(C=O)NR-, wherein R is selected from hydrogen, a C1-C6 linear or branched alkyl group.

[0101] 54. The compound according to item 53, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein L a is ethylene, linear propylene, linear butylene, linear pentylene, or -(CH2CH2O) n One, two or more of CH2CH2-, wherein n is 2, 3, 4, 5, 6, 7 or 8; Q a It is one, two, or more than three of -C(=O)-, -NR-, -NR(C=O)-, and -(C=O)NR-, wherein R is methyl and / or hydrogen.

[0102] 55. The compound according to item 53, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein:

[0103] L a is ethylene and / or -(CH2CH2O)4CH2CH2- or pentylene; Q aIt is -C(=O)-, -N(CH3)-, -NH(C=O)-, -C(=O)NH- or -C(=O)-N(CH3)-.

[0104] 56. The compound according to any one of items 1 to 55, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein L b is selected from a C1-C6 straight chain or branched chain alkylene group, a C2-C6 straight chain or branched chain alkenylene group, or a C2-C6 straight chain or branched chain alkynylene group, an arylene group having 6 to 10 carbon atoms in one or more rings, or a polyethylene glycol group, wherein the polyethylene glycol group is -(CH2CH2O) n -, -O(CH2CH2O) n -、-(CH2CH2O) n CH2-, -(CH2CH2O) n CH2CH2-, -CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n One or two or more of CH2-, n is an integer greater than or equal to 1, and the alkylene, alkenylene, and alkynylene groups are optionally substituted with substituents; Q b is selected from one, two or more of absent, -C(=O)NR-, -C(=O)O-, -OC(=O)NR-, -NRC(=O)-, -OC(=O)-, -NRC(=O)O-, -NRS(=O)2-, -NRC(=O)NR-, -C(=O)-, -OC(=O)O-, and R is selected from hydrogen, optionally substituted C1-C6 straight chain or branched alkyl.

[0105] 57. The compound according to item 56, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein L b is a C1-C6 straight or branched chain alkylene, or a C6-C 10 One or two or more than three arylene groups; Q b It is absent, one or two or more of -C(=O)-, -C(=O)NR-, -OC(=O)NR-, -NRC(=O)-, -NRC(=O)O-, or -NRC(=O)NR-, and R is selected from hydrogen, a C1-C6 straight chain or branched alkyl group.

[0106] 58. The compound according to item 56, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein L b Q is one or two or more of the group consisting of methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, sec-butylene, tert-butylene or benzylene; b is absent, one, two, or more than one of -C(=O)-, -NRC(=O)-, -NRC(=O)NR-, or -C(=O)NR-, and R is hydrogen or methyl.

[0107] 59. A compound according to any one of items 1 to 58, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the substituent is selected from deuterium, tritium, a halogen atom, an amino group, a hydroxyl group, a cyano group, a nitro group, an optionally substituted alkyl group, an alkenyl group, an alkynyl group, an optionally substituted heterocyclic group, an acyl group, an optionally substituted amino group, an optionally substituted carbamoyl group, an optionally substituted thiocarbamoyl group, an optionally substituted sulfamoyl group, an optionally substituted hydroxyl group, an optionally substituted sulfanyl group (SH) and an optionally substituted silyl group.

[0108] 60. The compound according to any one of items 1 to 59, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein the compound is selected from the following structures:

[0109] 61. A compound represented by formula (2), or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein the structure of formula (2) is as follows:

[0110] Ab comprises an antibody or an antigen-binding fragment thereof; M' is a linker; Sp is -L a -Q a -; A is selected from the group consisting of non-existent, hydrogen, and alkyl; T represents a linear or branched (C1-C8) trivalent alkyl group; Y, Y', and Y" are respectively -L b -Q b -;L a , L bQ is independently selected from absence, alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene, heteroarylene or polyethylene glycol group, wherein the alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene, heteroarylene or polyethylene glycol group is optionally substituted with a substituent; a , Q b are respectively selected from the group consisting of absence, -C(=O)NR-, -C(=O)O-, -OC(=O)NR-, -S(=O)2NR-, -NRC(=O)-, -OC(=O)-, -NRC(=O)O-, -NRS(=O)2-, -NRC(=O)NR-, -NHC-(=NH)NH-, -C(=O)-, -OC(=O)O-, -O-, -NR-, -S-, S(=O)2-, S(=O)-, or -Se-, wherein R is selected from hydrogen or an optionally substituted alkyl group; Z is selected from absence, hydrogen, an optionally substituted alkyl group, or a sulfonate inner salt; Z' and Z" are selected from absence, hydrogen, an optionally substituted alkyl group, a sulfonate inner salt, or a phosphate inner salt; at least one of Z, Z', and Z" is a sulfonate inner salt or a phosphate inner salt; X contains a self-destruct unit or does not exist; D comprises a drug molecule; i is an integer greater than or equal to 1; h is an integer from 1 to 6; o, p, and q are each an integer from 0 to 5; and j is an integer greater than or equal to 1.

[0111] 62. The compound according to item 61, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the formula (2) is formula (2-1):

[0112] wherein Ab, M', Sp, A, T, Y, h, o, i, j, X, D, and Z are as defined in item 61; and at least one of the i Zs is a sulfonate.

[0113] 63. The compound according to item 62, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the formula (2-1) is formula (2-1-1):

[0114] Wherein, AA is an amino acid residue, and W is an integer greater than or equal to 1.

[0115] 64. The compound according to item 61, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein the formula (2) is formula (2-2):

[0116] wherein Ab, M', Sp, A, T, Y, Z, o, p, i, h, j, X, Y', and D are as defined in item 61; and Z' is a sulfonic acid inner salt or a phosphate inner salt.

[0117] 65. The compound according to item 64, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the formula (2-2) is formula (2-2-1):

[0118] Wherein, AA is an amino acid residue, and W is an integer greater than or equal to 1.

[0119] 66. The compound according to item 61, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the formula (2) is formula (2-3):

[0120] wherein Ab, M', Sp, A, Y, T, Z, o, i, h, j, q, X, Y", and D are as defined in item 61; and Z" is a sulfonic acid inner salt or a phosphate inner salt.

[0121] 67. The compound according to item 66, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the formula (2-3) is formula (2-3-1):

[0122] Wherein, AA is an amino acid residue, and W is an integer greater than or equal to 1.

[0123] 68. The compound according to any one of items 61 to 67, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein, when A and Z are both absent, N, Y, and T form a ring, namely

[0124] Preferably

[0125] 69. A compound according to any one of items 63, 65 or 67, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the amino acid is selected from one or more of alanine, arginine, asparagine, aspartic acid, γ-carboxyglutamate, citrulline, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, norleucine, norvaline, ornithine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine.

[0126] 70. A compound according to any one of items 63, 65 or 67, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the amino acid is selected from one or more of valine, glutamic acid, citrulline, lysine, alanine, phenylalanine, arginine, glutamine, asparagine and glycine.

[0127] 71. A compound according to any one of items 63, 65 or 67, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the amino acid is selected from one or more of valine, citrulline, alanine, phenylalanine, glutamine and glycine.

[0128] 72. The compound according to any one of items 61 to 71, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein M' is selected from one of the following structures:

[0129] *Connect Ab.

[0130] 73. The compound according to any one of items 61 to 72, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein M' is selected from one of the following structures:

[0131] 74. The compound according to any one of items 61 to 73, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein:

[0132] M' is

[0133] 75. A compound according to any one of items 61 to 74, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein A is hydrogen, methyl, ethyl or propyl.

[0134] 76. A compound according to any one of items 61 to 75, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein A is hydrogen.

[0135] 77. The compound according to any one of items 61 to 76, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein T is one of the following structures:

[0136] 78. The compound according to any one of items 61 to 77, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein:

[0137] The T is

[0138] 79. The compound according to any one of items 61 to 78, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein L a One, two or more selected from alkylene, alkenylene, alkynylene, saturated or unsaturated alicyclic group having 3-10 ring carbon atoms, saturated or unsaturated heteroalicyclic group having 3-10 ring carbon atoms and 1 to 4 heteroatoms, arylene group having 6 to 18 carbon atoms in one or more rings, or heteroarylene group having 6 to 18 carbon atoms in one or more rings and 1 to 4 heteroatoms, or polyethylene glycol group, wherein the alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene group and heteroarylene group are optionally substituted by a substituent, and the heteroatom is selected from nitrogen, oxygen or sulfur.

[0139] 80. The compound according to any one of items 61 to 79, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein L a Selected from C1-C 10Straight or branched alkylene, C2-C 12 Straight or branched alkenylene, C2-C 12 One, two or more of a straight or branched alkynylene group, a saturated or unsaturated alicyclic group having 3-6 ring carbon atoms, a saturated or unsaturated heteroalicyclic group having 3-6 ring carbon atoms and 1 to 4 heteroatoms, or an arylene group having 6 to 10 carbon atoms in one or more rings, a heteroarylene group having 6 to 10 carbon atoms in one or more rings and 1 to 4 heteroatoms, or a polyethylene glycol group; the alkylene group, alkenylene group, alkynylene group, alicyclic group, heteroalicyclic group, arylene group and heteroarylene group are optionally substituted by a substituent, and the heteroatom is selected from nitrogen, oxygen or sulfur.

[0140] 81. The compound according to any one of items 61 to 80, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein L a One, two or more selected from C1-C6 straight chain or branched chain alkylene, C2-C6 straight chain or branched chain alkenylene, C2-C6 straight chain or branched chain alkynylene, or polyethylene glycol group, wherein the alkylene, alkenylene and alkynylene groups are optionally substituted by a substituent.

[0141] 82. The compound according to any one of items 61 to 81, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein L a is one, two or more of a C1-C6 straight or branched alkylene group or a polyethylene glycol group,

[0142] The polyethylene glycol group is -(CH2CH2O) n -, -O(CH2CH2O) n -、-(CH2CH2O) n CH2-, -(CH2CH2O) n CH2CH2-, -CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n CH2-, n is an integer greater than or equal to 1.

[0143] 83. The compound according to any one of items 61 to 82, or a tautomer, meso-racemate, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein L a is ethylene, linear propylene, linear butylene, linear pentylene, or -(CH2CH2O)n One, two or more of CH2CH2-, wherein n is 2, 3, 4, 5, 6, 7 or 8.

[0144] 84. The compound according to any one of items 61 to 83, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein L a It is ethylene and / or -(CH2CH2O)4CH2CH2- or pentylene.

[0145] 85. The compound according to any one of items 61 to 84, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein L b One, two or more than three selected from alkylene, alkenylene, alkynylene, saturated or unsaturated alicyclic group having 3-10 ring carbon atoms, saturated or unsaturated heteroalicyclic group having 3-10 ring carbon atoms and 1 to 4 heteroatoms, arylene group having 6 to 18 carbon atoms in one or more rings, or heteroarylene group having 6 to 18 carbon atoms in one or more rings and 1 to 4 heteroatoms, or polyethylene glycol, wherein the alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene group and heteroarylene group are optionally substituted by a substituent, and the heteroatom is selected from nitrogen, oxygen or sulfur.

[0146] 86. The compound according to any one of items 61 to 85, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein L b Selected from C1-C 10 Straight or branched alkylene, C2-C 12 Straight or branched alkenylene, C2-C 12 One or two or more of the group consisting of a straight or branched alkynylene group, a saturated or unsaturated alicyclic group having 3-6 ring carbon atoms, a saturated or unsaturated heteroalicyclic group containing 3-6 ring carbon atoms and 1 to 4 heteroatoms, or an arylene group having 6 to 10 carbon atoms in one or more rings, a heteroarylene group having 6 to 10 carbon atoms in one or more rings and 1 to 4 heteroatoms, or polyethylene glycol, wherein the alkylene group, alkenylene group, alkynylene group, alicyclic group, heteroalicyclic group, arylene group, and heteroarylene group are optionally substituted by a substituent, and the heteroatom is selected from nitrogen, oxygen, or sulfur.

[0147] 87. The compound according to any one of items 61 to 86, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein L b is selected from C1-C6 straight chain or branched chain alkylene, C2-C6 straight chain or branched chain alkenylene, or C2-C6 straight chain or branched chain alkynylene, or polyethylene glycol group, wherein the polyethylene glycol group is -(CH2CH2O) n -, -O(CH2CH2O) n -、-(CH2CH2O) n CH2-, -(CH2CH2O) n CH2CH2-, -CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n One, two, or more than three of CH2-, n is an integer greater than or equal to 1, and the alkylene, alkenylene, and alkynylene groups are optionally substituted with a substituent.

[0148] 88. The compound according to any one of items 61 to 87, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein L b is a C1-C6 straight or branched chain alkylene, or a C6-C 10 One or two or more than three arylene groups.

[0149] 89. The compound according to any one of items 61 to 88, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein L b It is one or two or more of the following: methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, sec-butylene, tert-butylene, or benzylene.

[0150] 90. The compound according to any one of items 61 to 89, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein Q a One, two or more selected from -C(=O)NR-, -NR-, -C(=O)O-, -OC(=O)NR-, -NRC(=O)-, -OC(=O)-, -NRC(=O)O-, -NRC(=O)NR-, NHC-(=NH)NH-, -C(=O)-, -OC(=O)O-, wherein R is selected from C1-C6 straight or branched chain alkyl optionally substituted with hydrogen.

[0151] 91. The compound according to item 90, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein:

[0152] Q a It is one, two, or more than three of -C(=O)-, -NR-, -C(=O)NR-, -NRC(=O)NR-, -OC(=O)O-, -OC(=O)NR-, -NRC(=O)-, or -OC(=O)-, wherein R is selected from hydrogen or an optionally substituted C1-C6 straight or branched alkyl group.

[0153] 92. The compound according to item 90, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein Q a It is one or two or more of -C(=O)-, -NR-, -NRC(=O)- and -(C=O)NR-, wherein R is selected from hydrogen or an optionally substituted C1-C6 linear or branched alkyl group.

[0154] 93. The compound according to item 90, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein Q a It is -C(=O)-, -N(CH3)-, -NHC(=O)-, -C(=O)-NH- or -C(=O)-N(CH3)-.

[0155] 94. The compound according to any one of items 61 to 93, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein Q b is selected from one, two or more of the group consisting of absent, -C(=O)NR-, -C(=O)O-, -OC(=O)NR-, -NRC(=O)-, -OC(=O)-, -NRC(=O)O-, -NRC(=O)NR-, NHC-(=NH)NH-, -C(=O)-, and -OC(=O)O-, wherein R is selected from hydrogen or an optionally substituted C1-C6 straight or branched alkyl group.

[0156] 95. The compound according to item 94, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein Q bis absent, one or two or more of -C(=O)-, -C(=O)NR-, -OC(=O)NR-, -NRC(=O)-, -NRC(=O)O-, or -NRC(=O)NR-, and R is selected from hydrogen or an optionally substituted C1-C6 linear or branched alkyl group.

[0157] 96. The compound according to item 94, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein Q b is absent, one, two, or more than one of -C(=O), -NRC(=O)-, -NRC(=O)NR-, or -C(=O)NR-, and R is hydrogen or methyl.

[0158] 97. The compound according to any one of items 61 to 96, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein X is selected from:

[0159] The right wavy line in X connects D; R4 is selected from hydrogen, C1-C6 straight or branched alkyl, -(CH2CH2O) m R2, -CH2CH2NR2R3, -CH2CH2SO2R2-, R2, R3 are independently selected from hydrogen, C1-C6 straight chain or branched alkyl, and m is an integer greater than 1.

[0160] 98. The compound according to item 97, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein:

[0161] X is

[0162] R4 is hydrogen, methyl, -CH2CH2OCH2CH2OH, -CH2CH2NMe2, -CH2CH2SO2Me.

[0163] 99. The compound according to item 97, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein:

[0164] X is R4 is hydrogen or methyl.

[0165] 100. The compound according to any one of items 61 to 99, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein the sulfonic acid inner salt is

[0166] A wavy line connects Y, Y' or Y";

[0167] wherein c, d, e, f, and g are all integers of 1, 2, 3, 4, 5, or 6;

[0168] R6, R7, R 11 、R 12 、R 13 、R 14 are independently selected from absence, hydrogen, and C1-C6 linear or branched alkyl.

[0169] 101. The compound according to item 100, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein the sulfonic acid inner salt is

[0170] Among them, R6, R7, R 11 、R 12 、R 13 and R 14 are selected from absence, methyl or ethyl, c, d, e, f, g are 1, 2 or 3 respectively.

[0171] 102. The compound according to item 100, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein the sulfonic acid inner salt is one of the following structures:

[0172] 103. The compound according to any one of items 61 to 102, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein the phosphate inner salt is

[0173] Wherein, a and b are integers of 1, 2, 3, 4, 5 or 6, R8, R9 and R 10 are independently selected from hydrogen, C1-C6 straight chain or branched alkyl.

[0174] 104. The compound according to item 103, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein the phosphate inner salt is

[0175] Among them, R8, R9, R 10 are methyl or ethyl respectively; a is 1 or 2 or 3; b is 1 or 2 or 3.

[0176] 105. The compound according to item 103, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein the phosphate inner salt is

[0177] Among them, R8, R9, R 10 All are methyl; a is 2; b is 1.

[0178] 106. A compound according to any one of items 61-105, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein D comprises a cytotoxin or an immunoagonist.

[0179] 107. A compound according to claim 106, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the cytotoxin is selected from one or two of a microtubule inhibitor, a DNA synthesis inhibitor, a topoisomerase inhibitor or an RNA polymerase 2 inhibitor.

[0180] 108. The compound according to item 106, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein the cytotoxin is selected from auristatins, maytansine alkaloids DMs, calicheamicin, duocarmycin, pyrrolobenzodiazepines One or more of camptothecin derivatives, camptothecin derivatives and α-amanitin.

[0181] 109. A compound according to claim 106, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the immune agonist is selected from a TLR7 / 8 agonist or a STING agonist.

[0182] 110. A compound according to any one of items 61-109, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein i is 2, 3, 4, 5, 6, 7 or 8.

[0183] 111. A compound according to any one of items 61-110, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein h is 1, 2, 3 or 4.

[0184] 112. A compound according to any one of items 61 to 111, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein o, p, q are 0, 1, 2 or 3 respectively.

[0185] 113. The compound according to any one of items 61 to 112, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein L a It is one, two or more of C1-C6 straight or branched alkylene groups, or polyethylene glycol groups, wherein the polyethylene glycol group is -(CH2CH2O) n -, -O(CH2CH2O) n -、-(CH2CH2O) n CH2-, -(CH2CH2O) n CH2CH2-, -CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n CH2-, n is an integer greater than or equal to 1; Q a It is one, two, or more than three of -C(=O)-, -NR(C=O)-, and -(C=O)NR-, wherein R is selected from hydrogen, a C1-C6 linear or branched alkyl group.

[0186] 114. The compound according to any one of items 61 to 113, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein L a is ethylene, linear propylene, linear butylene, linear pentylene, or -(CH2CH2O) n One, two or more of CH2CH2-, wherein n is 2, 3, 4, 5, 6, 7 or 8; Qa It is one, two, or more than three of -C(=O)-, -NR-, -NR(C=O)-, and -(C=O)NR-, wherein R is methyl and / or hydrogen.

[0187] 115. The compound according to item 114, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein:

[0188] L a is ethylene and / or -(CH2CH2O)4CH2CH2- or pentylene; Q a It is -C(=O)-, -N(CH3)-, -NH(C=O)-, -C(=O)NH- or -C(=O)-N(CH3)-.

[0189] 116. The compound according to any one of items 61 to 115, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein L b is selected from a C1-C6 straight chain or branched chain alkylene group, a C2-C6 straight chain or branched chain alkenylene group, or a C2-C6 straight chain or branched chain alkynylene group, an arylene group having 6 to 10 carbon atoms in one or more rings, or a polyethylene glycol group, wherein the polyethylene glycol group is -(CH2CH2O) n -, -O(CH2CH2O) n -、-(CH2CH2O) n CH2-, -(CH2CH2O) n CH2CH2-, -CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n One or two or more of CH2-, n is an integer greater than or equal to 1, and the alkylene, alkenylene, and alkynylene groups are optionally substituted with substituents; Q b is selected from one, two or more of absent, -C(=O)NR-, -C(=O)O-, -OC(=O)NR-, -NRC(=O)-, -OC(=O)-, -NRC(=O)O-, -NRS(=O)2-, -NRC(=O)NR-, -C(=O)-, -OC(=O)O-, and R is selected from hydrogen, optionally substituted C1-C6 straight chain or branched alkyl.

[0190] 117. The compound according to item 116, or its tautomer, meso-racemate, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein Lb is a C1-C6 straight or branched chain alkylene, or a C6-C 10 One or two or more than three arylene groups; Q b is absent, one or two or more of -C(=O)-, -C(=O)NR-, -OC(=O)NR-, -NRC(=O)-, -NRC(=O)O-, or -NRC(=O)NR-, and R is selected from hydrogen, an optionally substituted C1-C6 linear or branched alkyl group.

[0191] 118. The compound according to item 116, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein L b Q is one or two or more of the group consisting of methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, sec-butylene, tert-butylene or benzylene; b is absent, one, two, or more than one of -C(=O)-, -NRC(=O)-, -NRC(=O)NR-, or -C(=O)NR-, and R is hydrogen or methyl.

[0192] 119. A compound according to any one of items 61 to 118, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the substituent is selected from deuterium, tritium, a halogen atom, an amino group, a hydroxyl group, a cyano group, a nitro group, an optionally substituted alkyl group, an alkenyl group, an alkynyl group, an optionally substituted heterocyclic group, an acyl group, an optionally substituted amino group, an optionally substituted carbamoyl group, an optionally substituted thiocarbamoyl group, an optionally substituted sulfamoyl group, an optionally substituted hydroxyl group, an optionally substituted sulfanyl group (SH) and an optionally substituted silyl group.

[0193] 120. A compound according to any one of items 61 to 119, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein j is an integer from 1 to 20.

[0194] 121. The compound according to any one of items 61 to 120, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein the compound is selected from the following structures:

[0195] 122. A compound according to any one of items 61-121, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the antibody is selected from the following groups: murine antibody, chimeric antibody, humanized antibody and fully human antibody.

[0196] 123. A compound according to any one of items 61 to 122, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the antibody comprises a monoclonal antibody.

[0197] 124. A compound according to any one of items 61 to 123, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the antibody comprises a bispecific antibody.

[0198] 125. A compound according to any one of items 61-124, or its tautomer, racemate, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the antigen-binding fragment is selected from the following groups: Fab, Fab', Fv fragment, F(ab')2, F(ab)2, scFv, di-scFv and VHH.

[0199] 126. A compound according to any one of items 61 to 125, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein the Ab is selected from the following group: anti-claudin 18.2 antibody, anti-human folate receptor antibody, anti-HER2 antibody, anti-HER3 antibody, anti-TROP2 antibody, anti-B7-H3 antibody, anti-GPC20 antibody, anti-CDH6 antibody, anti-EGFR antibody, anti-EGFRvIII antibody, anti-AXL antibody, anti-Nectin-4 antibody, anti-Tissue factor antibody, anti-TIM-1 antibody, anti-PSMA antibody, anti-EpCAM antibody, anti-MUC1 antibody, anti-STEAP1 antibody, anti-GPNMB antibody, anti-FGF2 antibody, anti-FOLR1 antibody, anti-c-MET antibody, anti-GFR antibody, anti-AGS-16, anti-Guanylyl cyclase C antibody, anti-Mesothelin antibody, anti-SLC44A4 antibody, anti-EphA2 antibody, anti-AGS-5 antibody, anti-GPC-3 antibody, anti-c-KIT antibody, anti-ROR1 antibody, anti-PD-L1 antibody, anti-CD27L antibody, anti-5T4 antibody, anti-Mucin 16 antibody, anti-NaPi2b antibody, anti-STEAP antibody, anti-SLITRK6 antibody, anti-ETBR antibody, anti-BCMA antibody, anti-CEACAM5 antibody, anti-SC-16 antibody, anti-SLC39A6 antibody, anti-Delta-like protein3 antibody, anti-Claudin18.2 antibody, anti-CD19 antibody, anti-CD20 antibody, anti-CD22 antibody, anti-CD30 antibody, anti-CD33 antibody, anti-CD37 antibody, anti-CD45 antibody, anti-CD56 antibody, anti-CD66e antibody, anti-CD70 antibody, anti-CD73 antibody, anti-CD74 antibody, anti-CD79b antibody, anti-CD138 antibody, anti-CD147 antibody, anti-CD166 antibody, anti-CD223 antibody, anti-MUC16 antibody, anti-MSLN antibody, anti-ENPP3 antibody, anti-SLTRK6 antibody, anti-FGFR antibody, anti-LIV-1 antibody, anti-Lewis Y antibody, anti-av-integrin antibody, anti-ASCT2 antibody, and anti-C4.4a antibody, anti-CA-IX antibody, anti-CD324 antibody, anti-CD352 antibody, anti-CD44v6 antibody, anti-CD48a antibody, anti-CLL-1 antibody, anti-Cripto antibody, anti-CS1 antibody, anti-DPEP3 antibody, anti-Ephrin-A2 antibody, anti-Ephrin-A4 antibody, anti-ETBR antibody, anti-FGFR2 antibody, anti-FGFR3 antibody, anti-FLT3 antibody, anti-GD3 antibody, anti-GloboH antibody, anti-GPC3 antibody, anti-LAMP-1 antibody, anti-LRRC15 antibody, anti-Ly6E antibody, anti-MFI2 antibody, anti-NOTCH3 antibody, anti-p-cadherin antibody, anti-PRLR antibody and anti-RNF43 antibody, and antigen-binding fragments of the above antibodies.

[0200] 127. A compound according to any one of items 61 to 126, or its tautomer, racemate, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the heavy chain HCDR1, HCDR2 and HCDR3 and light chain LCDR1, LCDR2 and LCDR3 of Ab respectively contain the heavy chain HCDR1, HCDR2 and HCDR3 and light chain LCDR1, LCDR2 and LCDR3 of the antibody.

[0201] 128. A compound according to any one of items 61 to 127, or its tautomer, racemate, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the heavy chain variable region VH and light chain variable region VL of the Ab respectively comprise the heavy chain variable region VH and light chain variable region VL of the antibody.

[0202] 129. A compound according to any one of items 61 to 128, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the heavy chain and light chain of Ab respectively comprise the heavy chain and light chain of an antibody.

[0203] 130. A compound according to any one of items 61 to 129, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the Ab is an anti-claudin 18.2 antibody or an antigen-binding fragment thereof.

[0204] 131. A compound according to any one of items 61-130, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein the Ab is an anti-human folate receptor antibody or an antigen-binding fragment thereof.

[0205] 132. A pharmaceutical composition comprising a compound according to any one of items 1 to 131, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0206] 133. Use of a compound described in any one of items 1 to 132, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate in the preparation of a medicament for treating and / or preventing tumors.

[0207] 134. The use according to item 133, wherein the tumor is selected from tumors associated with the expression of the following group of targets: claudin 18.2, human folate receptor, HER2, HER3, TROP2, B7-H3, GPC20, CDH6, EGFR, EGFRvⅢ, AXL, Nectin-4, Tissue factor, TIM-1, PSMA, EpCAM, MUC1, STEAP1, GPNMB, FGF2, FOLR1, c-MET, GFR, AGS-16, Guanylyl cyclase C, Mesothelin, SLC44A4, EphA2, AGS-5, GPC-3, c-KIT, ROR1, PD-L1, CD27L, 5T4, Mucin 16, NaPi2b, STEAP, SLITRK6, ETBR, BCMA, CEACAM5, SC-16, SLC39A6, Delta-like protein3, Claudin18.2, CD19, CD20, CD22, CD30, CD33, CD37, CD45, CD56, CD66e, CD70, CD73, CD74, CD79b, CD138, CD147, CD166, CD223, MUC16, MSLN, ENPP3, SLTRK6, FGFR, LIV-1, Lewis Y, av-integrin, ASCT2, C4.4a, CA-IX, CD324, CD352, CD44v6, CD48a, CLL-1, Cripto, CS1, DPEP3, Ephrin-A2, Ephrin-A4, ETBR, FGFR2, FGFR3, FLT3, GD3, Globo H, GPC3, LAMP-1, LRRC15, Ly6E, MFI2, NOTCH3, p-cadherin, PRLR, and RNF43.

[0208] 135. The use according to item 133, wherein the tumors associated with the expression of the target include tumors with high expression of the target and / or target-positive tumors.

[0209] 136. The use according to item 133, wherein the tumor is selected from solid tumors, hematological tumors and metastatic, refractory or recurrent lesions of cancer.

[0210] 137. The use according to claim 133, wherein the tumor is selected from the group consisting of esophageal cancer, gastrointestinal cancer, pancreatic cancer, thyroid cancer, colorectal cancer, renal cancer, lung cancer (e.g., non-small cell lung cancer), liver cancer, gastric cancer, gastric adenocarcinoma, gastroesophageal junction (GEJ) adenocarcinoma, head and neck cancer, bladder cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, germ cell cancer, bone cancer, skin cancer, thymic cancer, bile duct cancer, gallbladder cancer, melanoma, mesothelioma, lymphoma, myeloma (e.g., multiple myeloma), sarcoma, glioblastoma, and leukemia.

[0211] The antibody drug conjugates provided by the present application, compared to the anionic and cationic hydrophilic groups of PEG, the fluorophore-antibody drug conjugates modified with water-soluble groups in the form of sulfonic acid inner salts greatly increase the specific accumulation at the tumor site. The anti-conjugated drug of the present application introduces a strongly hydrophilic sulfonic acid inner salt and / or phosphate inner salt side chain at the polypeptide (VC, VA, AAA, etc.)-PAB (self-destructive group) position of the linker. Since the sulfonic acid group or the phosphate group has both hydrogen bond donors and hydrogen bond acceptors, it belongs to a strongly hydrophilic group, and the sulfonic acid inner salt or the phosphate inner salt has both positive and negative charges, which can further increase the hydrophilicity. At the same time, the water-soluble group in the form of inner salts also has the potential to enhance the tumor targeting of the antibody drug conjugate and reduce nonspecific binding, thereby greatly enhancing the drugability (including hydrophilicity, stability) of the test antibody drug conjugate, reducing the drug clearance effect in the body, and improving the tumor treatment effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0212] The accompanying drawings are provided to facilitate a better understanding of the present application and do not constitute an undue limitation on the present application.

[0213] FIG1 is a molecular sieve exclusion chromatography (SEC) analysis diagram of the antibody drug conjugates of Comparative Example 2 and Experimental Example 6 provided in this application.

[0214] FIG2 is a molecular sieve exclusion chromatography (SEC) analysis diagram of the antibody drug conjugates of Comparative Example 3 and Experimental Example 9 provided in this application.

[0215] FIG3 is a molecular sieve exclusion chromatography (SEC) analysis chart of the antibody drug conjugates of Comparative Example 1 and Test Examples 1, 5a, 7, 9, and 11 provided in this application.

[0216] FIG4 is a hydrophobic interaction chromatography (HIC) analysis diagram of the antibody drug conjugates of Comparative Example 2 and Test Example 6 provided in this application.

[0217] FIG5 is a hydrophobic interaction chromatography (HIC) analysis diagram of the antibody drug conjugates of Comparative Example 3 and Test Example 9 provided in this application.

[0218] FIG6 is a hydrophobic interaction chromatography (HIC) analysis chart of the antibody drug conjugates of Comparative Example 1, Test Example 1, Test Example 5a, Test Example 7, Test Example 9, and Test Example 11 provided in this application.

[0219] Figure 7 is a deconvoluted mass spectrum of the light chain of Experimental Example 5a provided in this application.

[0220] FIG8 is a deconvoluted mass spectrum of the heavy chain of Experimental Example 5a provided in this application.

[0221] FIG9 is a reverse chromatogram of Experimental Example 5a provided in this application.

[0222] FIG10 is an in vitro cell killing curve of the 9D1 antibody-drug conjugate provided by the present application and a comparative sample on MC38-18.2 cells.

[0223] FIG11 is a tumor therapeutic effect curve of the 9D1 antibody-drug conjugate provided in the present application in the MC38-hClaudin18.2 mouse subcutaneous tumor model.

[0224] FIG12 is a tumor therapeutic effect curve of the 9D1 antibody-drug conjugate provided in the present application in the NUGC4-hClaudin18.2 mouse subcutaneous tumor model.

[0225] FIG13 is a tumor therapeutic effect curve of the Farletuzumab-LD-38 antibody drug conjugate provided in the present application in the OVCAR-3 mouse subcutaneous tumor model.

[0226] FIG14 is a tumor therapeutic effect curve of the Farletuzumab-LD-38 antibody drug conjugate provided in the present application on the IGROV1 mouse subcutaneous tumor model.

[0227] FIG15 is the pharmacokinetic curves of 9D1-LD-38 and 9D1-GGFG-Exatecan in rats provided in the present application.

[0228] FIG16 shows the cytotoxic activity of 9D1-LD-38, 9D1-LD-82, 9D1-LD-88 and 9D1-LD-89 against MC38-hClaudin18.2 cells.

[0229] FIG17 shows the results of immunogenic cell death induced by 9D1-LD-38 and 9D1-GGFG-Dxd in B16F10-hClaudin18.2 cells.

[0230] FIG18 shows the bystander effects of 9D1-LD-38 and 9D1-GGFG-Dxd on HEK293-hClaudin18.2 / HEK293 cells.

[0231] Figure 19 shows the results of releasing free toxin Exatecan after 9D1-LD-38 enzyme digestion.

[0232] FIG20 shows the in vitro serum incubation stability results of 9D1-LD-38, 9D1-LD-65, 9D1-LD-73, 9D1-LD-74, and 9D1-Deruxtecan (9D1-GGFG-Dxd).

[0233] FIG21 shows the changes in DAR values ​​of 9D1-LD-38, 9D1-LD-65, and 9D1-Deruxtecan (9D1-GGFG-Dxd) in vivo.

[0234] Figure 22 shows the contents of total toxin (bound toxin + free toxin) and free toxin in the plasma of tumor-bearing mice administered 9D1-LD-38, 9D1-LD-65, and 9D1-Deruxtecan (9D1-GGFG-Dxd).

[0235] Figure 23 shows the contents of total toxin (bound toxin + free toxin) and free toxin in the tissues of tumor-bearing mice administered 9D1-LD-38, 9D1-LD-65, and 9D1-Deruxtecan (9D1-GGFG-Dxd).

[0236] Figure 24 shows the pharmacodynamic results of 9D1-LD-38, 9D1-Deruxtecan (9D1-GGFG-Dxd) and 9D1-vc-PAB-MMAE in the p-gp-mediated drug-resistant tumor-bearing mouse model.

[0237] FIG25 shows the pharmacodynamic results of 9D1-LD-38 combined with ICI in an immunocompetent tumor-bearing mouse model.

[0238] FIG26 shows the results of the bystander effect of 9D1-LD-38 in tumor-bearing mice. DETAILED DESCRIPTION

[0239] The following description of exemplary embodiments of the present application includes various details of the embodiments of the present application to facilitate understanding, and should be considered as merely exemplary. Therefore, those skilled in the art will recognize that various changes and modifications may be made to the embodiments described herein without departing from the scope and spirit of the present application. Similarly, for the sake of clarity and conciseness, descriptions of well-known functions and structures are omitted in the following description.

[0240] Unless otherwise stated, some of the terms used in the specification and claims of this application are defined as follows:

[0241] "Alkyl" may be branched or unbranched, and preferably has 1 to about 12 carbon atoms. A more preferred class of alkyl groups has 1 to about 6 carbon atoms. Even more preferred are alkyl groups having 1, 2, 3, or 4 carbon atoms. Alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, etc. Alkyl groups may be substituted or unsubstituted.

[0242] "Alkenyl" is a branched or unbranched group having one or more double bonds and 2 to about 12 carbon atoms. A more preferred class of alkenyl groups has 2 to about 6 carbon atoms. Even more preferred are alkenyl groups having 2, 3, or 4 carbon atoms. Alkenyl groups include, but are not limited to, ethenyl, 1-propenyl, 2-propenyl, 1-methylethenyl, 1-butenyl, 2-butenyl, and 3-butenyl, and the like. Alkenyl groups can be substituted or unsubstituted.

[0243] "Alkynyl" is a branched or unbranched group having one or more triple bonds and 2 to about 12 carbon atoms. A more preferred class of alkynyl groups has 2 to about 6 carbon atoms, and even more preferred are alkynyl groups having 2, 3, or 4 carbon atoms. Alkynyl groups include, but are not limited to, acetylene, propyne, 1-butyne, and the like.

[0244] The "alicyclic group" refers to a cyclic substituent that does not have aromaticity, and may be a saturated hydrocarbon substituent or an unsaturated hydrocarbon substituent, for example, a cycloalkyl group, a cycloalkenyl group, or the like.

[0245] "Heteroalicyclic" refers to a substituent group formed by a heterocyclic compound containing at least one heteroatom in the ring backbone and lacking aromaticity. The heteroatom can be oxygen, nitrogen, sulfur atoms, etc., and includes monocyclic, polycyclic, condensed, bridged, and spirocyclic rings. For example, it has a 5- to 7-membered monocyclic ring or a 7- to 10-membered bicyclic or tricyclic ring, which may contain 1, 2, or 3 atoms selected from nitrogen, oxygen, and / or sulfur. Heteroalicyclic groups include, but are not limited to, morpholinyl, oxetanyl, thiomorpholinyl, tetrahydrofuranyl, tetrahydropyranyl, 1,1-dioxo-thiomorpholinyl, piperidinyl, 2-oxo-piperidinyl, pyrrolidinyl, 2-oxo-pyrrolidinyl, piperazin-2-one, 8-oxa-3-aza-bicyclo[3.2.1]octyl, piperazinyl, and hexahydropyrimidine. The heterocyclic group can be substituted or unsubstituted.

[0246] "Aryl" refers to a carbocyclic aromatic system containing one or two rings, wherein the rings may be fused together. The term "aryl" includes monocyclic or bicyclic aromatic groups, such as phenyl, naphthyl, and tetrahydronaphthyl aromatic groups. Preferred aryl groups are C6-C10 aromatic groups, more preferably phenyl and naphthyl, and most preferably naphthyl. Aryl groups may be substituted or unsubstituted.

[0247] "Heteroaryl" refers to an aromatic 5-6 membered monocyclic or 8-10 membered bicyclic ring which may contain 1 to 4 atoms selected from nitrogen, oxygen and / or sulfur. Examples of "heteroaryl" include, but are not limited to, furyl, pyridyl, 2-oxo-1,2-dihydropyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, thienyl, isoxazolyl, oxazolyl, oxadiazolyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, benzodioxolyl, benzothienyl, benzimidazolyl, indolyl, isoindolyl, 1,3-dioxo-isoindolyl, quinolyl, indazolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl. Heteroaryl may be substituted or unsubstituted.

[0248] "Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3 hydrogen atoms, in a group are replaced independently of one another by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and a person skilled in the art can determine (by experiment or theory) which substitutions are possible or impossible without undue effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated (e.g., olefinic) bond.

[0249] The term "substituted" or "substituted" as used herein, unless otherwise specified, means that a group may be substituted by one or more groups selected from the group consisting of alkyl, alkoxy, alkylthio, alkylamino, halogen, thio, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, amino, haloalkyl, and hydroxyalkyl;

[0250] Whenever a group is described as "optionally substituted with substituents", the group may be unsubstituted or substituted with one or more of the substituents indicated. Similarly, when a group is described as "unsubstituted or substituted", if substituted, the substituents may be selected from one or more of the substituents indicated. If no substituents are indicated, it means that the group shown as "optionally substituted with substituents" or "substituted" may be substituted with one or more groups individually or independently selected from alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, heteroalicyclic, aralkyl, heteroaralkyl, (heteroalicyclic)alkyl, hydroxy, protected hydroxy, alkoxy, aryloxy, acyl, sulfhydryl, alkylthio, arylthio, cyano, halogen, thiocarbonyl, O-carbamyl, N- -carbamoyl, O-thiocarbamoyl, N-thiocarbamoyl, C-amide, N-amide, S-sulfonamide, N-sulfonamide, C-carboxyl, protected C-carboxyl, O-carboxyl, isocyanate, thiocyanate, isothiocyanate, nitro, silyl, sulfo, sulfinyl, sulfonyl, haloalkyl, haloalkoxy, trihalomethanesulfonyl, trihalomethanesulfonamido, amino, monosubstituted amino and disubstituted amino groups and protected derivatives thereof.

[0251] Halogen substituents include F, Cl, Br, and I. "Hydroxyl" refers to an -OH group. "Halogen" refers to fluorine, chlorine, bromine, and iodine.

[0252] "Amino" refers to -NH2. "Cyano" refers to -CN. "Nitro" refers to -NO2. "Carboxyl" refers to -C(O)OH. "Acyl" refers to -C(O)R.

[0253] The term "hydrogen" or "H" in the chemical structures used herein, unless otherwise indicated, should be understood to include not only 1 H, also includes deuterium ( 2 H, D), tritium ( 3 "Deuterated isotope derivative" refers to a compound in which one or more H atoms are replaced by D atoms to obtain the corresponding compound.

[0254] A "pharmaceutical composition" refers to a mixture containing one or more compounds described herein, or their physiologically acceptable salts or prodrugs, together with other chemical components, as well as other components such as physiologically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitating absorption of the active ingredients and thereby exerting their biological activity.

[0255] The present application proposes a compound represented by formula (1), or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein the structure of formula (1) is shown below,

[0256] M is a linking group; Sp is -L a -Q a -; A is selected from the group consisting of non-existent, hydrogen, and alkyl; T represents a linear or branched (C1-C8) trivalent alkyl group; Y, Y', and Y" are respectively -L b -Q b -;L a , L b Q is independently selected from absence, alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene, heteroarylene or polyethylene glycol group, wherein the alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene, heteroarylene or polyethylene glycol group is optionally substituted with a substituent; a , Q b are respectively selected from the group consisting of absence, -C(=O)NR-, -C(=O)O-, -OC(=O)NR-, -S(=O)2NR-, -NRC(=O)-, -OC(=O)-, -NRC(=O)O-, -NRS(=O)2-, -NRC(=O)NR-, -NHC-(=NH)NH-, -C(=O)-, -OC(=O)O-, -O-, -NR-, -S-, S(=O)2-, S(=O)-, or -Se-, wherein R is selected from hydrogen or an optionally substituted alkyl group; Z is selected from absence, hydrogen, an optionally substituted alkyl group, or a sulfonate inner salt; Z' and Z" are selected from absence, hydrogen, an optionally substituted alkyl group, a sulfonate inner salt, or a phosphate inner salt; at least one of Z, Z', and Z" is a sulfonate inner salt or a phosphate inner salt; X comprises a self-destructing unit or is absent; D comprises a drug molecule; i is an integer greater than or equal to 1, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, etc.; h is an integer from 1 to 6, for example, 1, 2, 3, 4, 5, 6, etc.; o, p, and q are each an integer from 0 to 5, for example, 0, 1, 2, 3, 4, 5, etc.

[0257] When h is greater than 1, multiple Sp can have different structures or the same structure, that is, multiple L a It can be of different structures or the same structure; multiple Q a It can be of different structures or the same structure. a and Q a The number can be the same or different.

[0258] When i is greater than 1, multiple A's may be different or the same, multiple T's may be different or the same, multiple Y's may be different or the same, and multiple Z's may be different or the same.

[0259] When p is greater than 1, the multiple Y's may have different structures or the same structure.

[0260] When q is greater than 1, the multiple Y″ may have different structures or the same structure.

[0261] In this application, the formula (1) is formula (1-1):

[0262] Wherein, M, Sp, A, T, Y, h, o, i, X, D, and Z are as defined above; at least one of the i Zs is a sulfonate.

[0263] In this application, the formula (1-1) is formula (1-1-1):

[0264] Wherein, AA is an amino acid residue, and w is an integer greater than or equal to 1, for example, 1, 2, 3, 4, 5, 6, etc. When w is greater than 1, multiple AAs may be residues of the same amino acid or residues of different amino acids.

[0265] In this application, the formula (1) is the formula (1-2):

[0266] Wherein, M, Sp, A, T, Y, Z, o, p, i, h, X, Y′, and D are as defined above; and Z′ is a sulfonic acid inner salt or a phosphate inner salt.

[0267] In this application, the formula (1) is formula (1-2-1):

[0268] Wherein, AA is an amino acid residue, and w is an integer greater than or equal to 1, for example, 1, 2, 3, 4, 5, 6, etc. When w is greater than 1, multiple AAs can be residues of the same amino acid or residues of different amino acids.

[0269] In this application, the formula (1) is formula (1-3):

[0270] Wherein, M, Sp, A, Y, T, Z, o, i, h, q, X, Y″, and D are as defined above; Z″ is a sulfonic acid inner salt or a phosphate inner salt.

[0271] In the present application, when Z is hydrogen or an optionally substituted alkyl group, It is an amino acid residue, also called AA.

[0272] In the present application, when Z is a sulfonic acid inner salt, for

[0273] In this application, when A and Z are absent, N, Y, and T form a ring. for Preferably

[0274] In the present application, when Z is selected from hydrogen or an optionally substituted alkyl group, the formula (1) is formula (1-3-1):

[0275] Wherein, AA is an amino acid residue, and w is an integer greater than or equal to 1, for example, 1, 2, 3, 4, 5, 6, etc. When w is greater than 1, multiple AAs can be residues of the same amino acid or residues of different amino acids.

[0276] In the present application, the amino acid is selected from one or more of alanine, arginine, asparagine, aspartic acid, γ-carboxyglutamate, citrulline, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, norleucine, norvaline, ornithine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine.

[0277] In the present application, the amino acid is selected from one or more of valine, glutamic acid, citrulline, lysine, alanine, phenylalanine, arginine, glutamine, asparagine and glycine.

[0278] In the present application, the amino acid is selected from one or more of valine, citrulline, alanine, phenylalanine, glutamine and glycine.

[0279] When w is greater than 1, multiple AAs can be the same or different, as follows:

[0280] In some embodiments, when w is 2, the two AAs may be independently a valine residue and a citrulline residue, respectively.

[0281] In some embodiments, when w is greater than 2, the w AAs may be independently a valine residue and a citrulline residue, wherein the number of valine residues may be 1 or 2 or more, or the number of citrulline residues may be 1 or 2 or more. For example, when w is 3, the three AAs are respectively two valine residues and one citrulline residue. For another example, when w is 3, the three AAs are respectively one valine residue and two citrulline residues.

[0282] In some embodiments, when w is 2, the two AAs may be independently a valine residue and an alanine residue, respectively.

[0283] In some embodiments, when w is greater than 2, the w AAs may be independently valine residues and alanine residues, wherein the number of valine residues may be 1 or 2 or more, or the number of alanine residues may be 1 or 2 or more. For example, when w is 3, the three AAs are respectively two valine residues and one alanine residue. For another example, when w is 3, the three AAs are respectively one valine residue and two alanine residues.

[0284] In some embodiments, when w is 2, the two AAs may be independently a valine residue and a phenylalanine residue, respectively.

[0285] In some embodiments, when w is greater than 2, the w AAs may be independently a valine residue and a phenylalanine residue, wherein the number of valine residues may be 1 or 2 or more, or the number of phenylalanine residues may be 1 or 2 or more. For example, when w is 3, the three AAs are respectively two valine residues and one phenylalanine residue. For another example, when w is 3, the three AAs are respectively one valine residue and two phenylalanine residues.

[0286] In some embodiments, when w is 2, the two AAs may be independently a valine residue and a glutamine residue, respectively.

[0287] In some embodiments, when w is greater than 2, the w AAs may be independently a valine residue and a glutamine residue, wherein the number of valine residues may be 1 or 2 or more, or the number of glutamine residues may be 1 or 2 or more. For example, when w is 3, the three AAs are respectively two valine residues and one glutamine residue. For another example, when w is 3, the three AAs are respectively one valine residue and two glutamine residues.

[0288] In some embodiments, when w is 2, the two AAs may be independently a valine residue and a glycine residue, respectively.

[0289] In some embodiments, when w is greater than 2, the w AAs may be independently a valine residue and a glycine residue, wherein the number of valine residues may be 1 or 2 or more, or the number of glycine residues may be 1 or 2 or more. For example, when w is 3, the three AAs are respectively two valine residues and one glycine residue. For another example, when w is 3, the three AAs are respectively one valine residue and two glycine residues.

[0290] In some embodiments, when w is 2, the two AAs may be independently a citrulline residue and an alanine residue, respectively.

[0291] In some embodiments, when w is greater than 2, the w AAs may be independently citrulline residues and alanine residues, wherein the number of citrulline residues may be 1 or 2 or more, or the number of alanine residues may be 1 or 2 or more. For example, when w is 3, the three AAs are respectively two citrulline residues and one alanine residue. For another example, when w is 3, the three AAs are respectively one citrulline residue and two alanine residues.

[0292] In some embodiments, when w is greater than 1, the w AAs may be independently citrulline residues and phenylalanine residues, respectively, wherein the number of citrulline residues may be 1 or 2 or more, or the number of phenylalanine residues may be 1 or 2 or more.

[0293] In some embodiments, when w is greater than 1, the w AAs may be independently citrulline residues and glutamine residues, respectively, wherein the number of citrulline residues may be 1 or 2 or more, or the number of glutamine residues may be 1 or 2 or more.

[0294] In some embodiments, when w is greater than 1, the w AAs may be independently citrulline residues and glycine residues, respectively, wherein the number of citrulline residues may be 1 or 2 or more, or the number of glycine residues may be 1 or 2 or more.

[0295] In some embodiments, when w is greater than 1, the w AAs may be independently alanine residues and phenylalanine residues, respectively, wherein the number of alanine residues may be 1 or more than 2 or the number of phenylalanine residues may be 1 or more than 2.

[0296] In some embodiments, when w is greater than 1, the w AAs may be independently alanine residues and glutamine residues, respectively, wherein the number of alanine residues may be 1 or 2 or more, or the number of glutamine residues may be 1 or 2 or more.

[0297] In some embodiments, when w is greater than 1, the w AAs may be independently alanine residues and glycine residues, respectively, wherein the number of alanine residues may be 1 or 2 or more, or the number of glycine residues may be 1 or 2 or more.

[0298] In some embodiments, when w is greater than 1, the w AAs may be independently phenylalanine residues and glutamine residues, respectively, wherein the number of phenylalanine residues may be 1 or 2 or more, or the number of glutamine residues may be 1 or 2 or more.

[0299] In some embodiments, when w is greater than 1, the w AAs may be independently phenylalanine residues and glycine residues, respectively, wherein the number of phenylalanine residues may be 1 or 2 or more, or the number of glycine residues may be 1 or 2 or more.

[0300] In some embodiments, when w is greater than 1, the w AAs may be independently glutamine residues and glycine residues, respectively, wherein the number of glutamine residues may be 1 or 2 or more, or the number of glycine residues may be 1 or 2 or more.

[0301] In some embodiments, when w is greater than 1, the w AAs may be independently valine residues, citrulline residues, and alanine residues, respectively, wherein the number of valine residues may be 1 or more, the number of citrulline residues may be 1 or more, or the number of alanine residues may be 1 or more.

[0302] In some embodiments, when w is greater than 1, the w AAs may be independently a valine residue, a citrulline residue, and a phenylalanine residue, respectively, wherein the number of valine residues may be 1 or more, the number of citrulline residues may be 1 or more, or the number of phenylalanine residues may be 1 or more.

[0303] In some embodiments, when w is greater than 1, the w AAs may be independently a valine residue, a citrulline residue, and a glutamine residue, respectively, wherein the number of valine residues may be 1 or more, the number of citrulline residues may be 1 or more, or the number of glutamine residues may be 1 or more.

[0304] In some embodiments, when w is greater than 1, the w AAs may be independently a valine residue, a citrulline residue, and a glycine residue, respectively, wherein the number of valine residues may be 1 or more, the number of citrulline residues may be 1 or more, or the number of glycine residues may be 1 or more.

[0305] In some embodiments, when w is greater than 1, the w AAs may be independently a valine residue, an alanine residue, and a phenylalanine residue, respectively, wherein the number of valine residues may be 1 or more, the number of alanine residues may be 1 or more, or the number of phenylalanine residues may be 1 or more.

[0306] In some embodiments, when w is greater than 1, the w AAs may be independently a valine residue, an alanine residue, and a glutamine residue, respectively, wherein the number of valine residues may be 1 or more, the number of alanine residues may be 1 or more, or the number of glutamine residues may be 1 or more.

[0307] In some embodiments, when w is greater than 1, the w AAs may be independently a valine residue, an alanine residue, and a glycine residue, respectively, wherein the number of valine residues may be 1 or more, the number of alanine residues may be 1 or more, or the number of glycine residues may be 1 or more.

[0308] In some embodiments, when w is greater than 1, the w AAs may be independently a valine residue, a phenylalanine residue, and a glutamine residue, respectively, wherein the number of valine residues may be 1 or more, the number of phenylalanine residues may be 1 or more, or the number of glutamine residues may be 1 or more.

[0309] In some embodiments, when w is greater than 1, the w AAs may be independently a valine residue, a phenylalanine residue, and a glycine residue, respectively, wherein the number of valine residues may be 1 or more, the number of phenylalanine residues may be 1 or more, or the number of glycine residues may be 1 or more.

[0310] In some embodiments, when w is greater than 1, the w AAs may be independently a valine residue, a glutamine residue, and a glycine residue, respectively, wherein the number of valine residues may be 1 or more, the number of glutamine residues may be 1 or more, or the number of glycine residues may be 1 or more.

[0311] In some embodiments, when w is greater than 1, the w AAs may be independently a citrulline residue, an alanine residue, and a phenylalanine residue, respectively, wherein the number of citrulline residues may be 1 or more, the number of alanine residues may be 1 or more, or the number of phenylalanine residues may be 1 or more.

[0312] In some embodiments, when w is greater than 1, the w AAs may be independently a citrulline residue, an alanine residue, and a glutamine residue, respectively, wherein the number of citrulline residues may be 1 or more, the number of alanine residues may be 1 or more, or the number of glutamine residues may be 1 or more.

[0313] In some embodiments, when w is greater than 1, the w AAs may be independently a citrulline residue, an alanine residue, and a glycine residue, respectively, wherein the number of citrulline residues may be 1 or more, the number of alanine residues may be 1 or more, or the number of glycine residues may be 1 or more.

[0314] In some embodiments, when w is greater than 1, the w AAs may be independently an alanine residue, a phenylalanine residue, and a glutamine residue, respectively, wherein the number of alanine residues may be 1 or more, the number of phenylalanine residues may be 1 or more, or the number of glutamine residues may be 1 or more.

[0315] In some embodiments, when w is greater than 1, the w AAs may be independently an alanine residue, a phenylalanine residue, and a glycine residue, respectively, wherein the number of alanine residues may be 1 or more than 2, the number of phenylalanine residues may be 1 or more than 2, or the number of glycine residues may be 1 or more than 2.

[0316] In some embodiments, when w is greater than 1, the w AAs may be independently a phenylalanine residue, a glutamine residue, and a glycine residue, respectively, wherein the number of phenylalanine residues may be 1 or more than 2, the number of glutamine residues may be 1 or more than 2, or the number of glycine residues may be 1 or more than 2.

[0317] In some embodiments, when w is greater than 1, the w AAs may be independently a valine residue, a citrulline residue, an alanine residue, and a phenylalanine residue, respectively, wherein the number of valine residues may be 1 or more, the number of citrulline residues may be 1 or more, the number of alanine residues may be 1 or more, or the number of phenylalanine residues may be 1 or more.

[0318] In some embodiments, when w is greater than 1, the w AAs may be independently a valine residue, a citrulline residue, an alanine residue, and a glutamine residue, respectively, wherein the number of valine residues may be 1 or more, the number of citrulline residues may be 1 or more, the number of alanine residues may be 1 or more, or the number of glutamine residues may be 1 or more.

[0319] In some embodiments, when w is greater than 1, the w AAs may be independently a valine residue, a citrulline residue, an alanine residue, and a glycine residue, respectively, wherein the number of valine residues may be 1 or more, the number of citrulline residues may be 1 or more, the number of alanine residues may be 1 or more, or the number of glycine residues may be 1 or more.

[0320] In some embodiments, when w is greater than 1, the w AAs may be independently a citrulline residue, an alanine residue, a phenylalanine residue, and a glutamine residue, respectively, wherein the number of citrulline residues may be 1 or more, the number of alanine residues may be 1 or more, the number of phenylalanine residues may be 1 or more, or the number of glutamine residues may be 1 or more.

[0321] In some embodiments, when w is greater than 1, the w AAs may be independently a citrulline residue, an alanine residue, a phenylalanine residue, and a glycine residue, respectively, wherein the number of citrulline residues may be 1 or more, the number of alanine residues may be 1 or more, the number of phenylalanine residues may be 1 or more, or the number of glycine residues may be 1 or more.

[0322] In some embodiments, when w is greater than 1, the w AAs may be independently an alanine residue, a phenylalanine residue, a glutamine residue, and a glycine residue, respectively, wherein the number of alanine residues may be 1 or more, the number of phenylalanine residues may be 1 or more, the number of glutamine residues may be 1 or more, or the number of glycine residues may be 1 or more.

[0323] In some embodiments, when w is greater than 1, the w AAs may be independently a valine residue, a citrulline residue, an alanine residue, a phenylalanine residue, and a glutamine residue, wherein the number of valine residues may be 1 or more, the number of citrulline residues may be 1 or more, the number of alanine residues may be 1 or more, the number of phenylalanine residues may be 1 or more, or the number of glutamine residues may be 1 or more.

[0324] In some embodiments, when w is greater than 1, the w AAs may be independently a valine residue, a citrulline residue, an alanine residue, a phenylalanine residue, and a glycine residue, respectively, wherein the number of valine residues may be 1 or more, the number of citrulline residues may be 1 or more, the number of alanine residues may be 1 or more, the number of phenylalanine residues may be 1 or more, or the number of glycine residues may be 1 or more.

[0325] In some embodiments, when w is greater than 1, the w AAs may be independently a citrulline residue, an alanine residue, a phenylalanine residue, a glutamine residue, and a glycine residue, respectively, wherein the number of citrulline residues may be 1 or more, the number of alanine residues may be 1 or more, the number of phenylalanine residues may be 1 or more, the number of glutamine residues may be 1 or more, or the number of glycine residues may be 1 or more.

[0326] In the present application, M is N3, SH, ONH2, NH2, CO2H, (alkyl)-CO-, HCO-, BrCH 2- 、ICH2-、

[0327] Furthermore, M is

[0328] In some embodiments, M is

[0329] In some embodiments, M is

[0330] In some embodiments, M is

[0331] In some embodiments, M is

[0332] In the present application, A is hydrogen, methyl, ethyl or propyl.

[0333] In some embodiments, A is hydrogen. In some embodiments, A is methyl. In some embodiments, A is ethyl. In some embodiments, A is propyl.

[0334] In this application, T is one of the following structures:

[0335] In some embodiments, T is

[0336] In this application, La One, two or more selected from alkylene, alkenylene, alkynylene, saturated or unsaturated alicyclic group having 3-10 ring carbon atoms, saturated or unsaturated heteroalicyclic group having 3-10 ring carbon atoms and 1 to 4 heteroatoms, arylene group having 6 to 18 carbon atoms in one or more rings, or heteroarylene group having 6 to 18 carbon atoms in one or more rings and 1 to 4 heteroatoms, or polyethylene glycol group, wherein the alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene group and heteroarylene group are optionally substituted by a substituent, and the heteroatom is selected from nitrogen, oxygen or sulfur.

[0337] Furthermore, L a Selected from C1-C 10 Straight or branched alkylene, C2-C 12 Straight or branched alkenylene, C2-C 12 One, two or more of a straight or branched alkynylene group, a saturated or unsaturated alicyclic group having 3-6 ring carbon atoms, a saturated or unsaturated heteroalicyclic group having 3-6 ring carbon atoms and 1 to 4 heteroatoms, or an arylene group having 6 to 10 carbon atoms in one or more rings, a heteroarylene group having 6 to 10 carbon atoms in one or more rings and 1 to 4 heteroatoms, or a polyethylene glycol group, wherein the alkylene group, alkenylene group, alkynylene group, alicyclic group, heteroalicyclic group, arylene group and heteroarylene group are optionally substituted by a substituent, wherein the heteroatom is selected from nitrogen, oxygen or sulfur. Further, L a One, two or more selected from C1-C6 straight chain or branched chain alkylene, C2-C6 straight chain or branched chain alkenylene, C2-C6 straight chain or branched chain alkynylene, or polyethylene glycol group, wherein the alkylene, alkenylene and alkynylene groups are optionally substituted by a substituent.

[0338] Furthermore, L a It is one, two or more of C1-C6 straight or branched alkylene groups, or polyethylene glycol groups, wherein the polyethylene glycol group is -(CH2CH2O) n -, -O(CH2CH2O) n -、-(CH2CH2O) n CH2-, -(CH2CH2O) n CH2CH2-, -CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n CH2-, n is an integer greater than or equal to 1.

[0339] Furthermore, L a is ethylene, linear propylene, linear butylene, linear pentylene, or -(CH2CH2O)n One, two or more of CH2CH2-, wherein n is 2, 3, 4, 5, 6, 7 or 8.

[0340] Furthermore, L a It is ethylene and / or -(CH2CH2O)4CH2CH2- or pentylene.

[0341] In some embodiments, L a The number of them is 2, one of which is ethylene and the other is -(CH2CH2O)4CH2CH2-.

[0342] In some embodiments, L a When the number of is 1, it is ethylene. a When the number of is 1, it is pentylene. In some embodiments, L a When the number is 1, it is -(CH2CH2O)4CH2CH2-.

[0343] In this application, L b One or two or more selected from the group consisting of an alkylene group, an alkenylene group, an alkynylene group, a saturated or unsaturated alicyclic group having 3-10 ring carbon atoms, a saturated or unsaturated heteroalicyclic group having 3-10 ring carbon atoms and 1 to 4 heteroatoms, an arylene group having 6 to 18 carbon atoms in one or more rings, or a heteroarylene group having 6 to 18 carbon atoms in one or more rings and 1 to 4 heteroatoms, or polyethylene glycol,

[0344] The alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene group, and heteroarylene group are optionally substituted with a substituent, and the heteroatom is selected from nitrogen, oxygen, or sulfur.

[0345] Furthermore, L b Selected from C1-C 10 Straight or branched alkylene, C2-C 12 Straight or branched alkenylene, C2-C 12 One or more of a straight or branched alkynylene group, a saturated or unsaturated alicyclic group having 3 to 6 ring carbon atoms, a saturated or unsaturated heteroalicyclic group having 3 to 6 ring carbon atoms and 1 to 4 heteroatoms, an arylene group having 6 to 10 carbon atoms in one or more rings, a heteroarylene group having 6 to 10 carbon atoms in one or more rings and 1 to 4 heteroatoms, or polyethylene glycol.

[0346] The alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene group, and heteroarylene group are optionally substituted with a substituent, and the heteroatom is selected from nitrogen, oxygen, or sulfur.

[0347] Furthermore, L b is selected from C1-C6 straight chain or branched chain alkylene, C2-C6 straight chain or branched chain alkenylene, or C2-C6 straight chain or branched chain alkynylene, or polyethylene glycol group,

[0348] The polyethylene glycol group is -(CH2CH2O) n -, -O(CH2CH2O) n -、-(CH2CH2O) n CH2-, -(CH2CH2O) n CH2CH2-, -CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n One, two, or more than three of CH2-, n is an integer greater than or equal to 1, and the alkylene, alkenylene, and alkynylene groups are optionally substituted with a substituent.

[0349] Furthermore, L b is a C1-C6 straight or branched chain alkylene, or a C6-C 10 One or two or more than three arylene groups.

[0350] Furthermore, L b It is one or two or more of the following: methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, sec-butylene, tert-butylene, or benzylene.

[0351] When L b When the number is multiple, multiple L b It can be the same structure or different structures, as shown below:

[0352] In some embodiments, L b When the number is 1 or more, L b In some embodiments, L b When the number is 1 or more, L b In some embodiments, L b When the number is 1 or more, L b In some embodiments, L b When the number is 1 or more, L b In some embodiments, L b When the number is 1 or more, L b In some embodiments, L b When the number is 1 or more, L b In some embodiments, L bWhen the number is 1 or more, L b In some embodiments, L b When the number is 1 or more, L b In some embodiments, L b When the number is 1 or more, L b All are benzylene.

[0353] In some embodiments, L b When the number is multiple, multiple L b The number of methylene groups can be 1 or more than 2, and the number of ethylene groups can be 1 or more than 2, for example, L b When the number is 3, 3 L b There are two methylene groups and one ethylene group respectively. b When the number is 3, 3 L b One methylene and two ethylene groups respectively.

[0354] In some embodiments, L b When the number is multiple, multiple L b The number of methylene groups and n-propylene groups can be 1 or more, and the number of n-propylene groups can be 1 or more, for example, L b When the number is 3, 3 L b There are two methylene groups and one n-propylene group respectively. For example, L b When the number is 3, 3 L b There are 1 methylene and two n-propylene groups respectively.

[0355] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are methylene and isopropylene, wherein the number of methylene can be 1 or more than 2, and the number of isopropylene can be 1 or more than 2.

[0356] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are methylene and n-butylene, wherein the number of methylene can be 1 or more than 2, and the number of n-butylene can be 1 or more than 2.

[0357] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are methylene and isobutylene groups, wherein the number of methylene groups can be 1 or more than 2, and the number of isobutylene groups can be 1 or more than 2.

[0358] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are methylene and sec-butylene groups, wherein the number of methylene groups can be 1 or more than 2, and the number of sec-butylene groups can be 1 or more than 2.

[0359] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are methylene and tert-butylene groups, wherein the number of methylene groups can be 1 or more than 2, and the number of tert-butylene groups can be 1 or more than 2.

[0360] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are methylene and benzylene groups, wherein the number of methylene groups may be 1 or more than 2, and the number of benzylene groups may be 1 or more than 2.

[0361] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are ethylene and n-propylene, wherein the number of ethylene can be 1 or more than 2, and the number of n-propylene can be 1 or more than 2.

[0362] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are ethylene and isopropylene, wherein the number of ethylene can be 1 or more than 2, and the number of isopropylene can be 1 or more than 2.

[0363] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are ethylene and n-butylene, wherein the number of ethylene can be 1 or more than 2, and the number of n-butylene can be 1 or more than 2.

[0364] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are ethylene and isobutylene, wherein the number of ethylene can be 1 or more than 2, and the number of isobutylene can be 1 or more than 2.

[0365] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are ethylene and sec-butylene, wherein the number of ethylene can be 1 or more than 2, and the number of sec-butylene can be 1 or more than 2.

[0366] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are ethylene and tert-butylene groups, wherein the number of ethylene groups can be 1 or more than 2, and the number of tert-butylene groups can be 1 or more than 2.

[0367] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are ethylene and benzylene, wherein the number of ethylene can be 1 or more than 2, and the number of benzylene can be 1 or more than 2.

[0368] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are n-propylene and isopropylene, wherein the number of n-propylene can be 1 or more than 2, and the number of isopropylene can be 1 or more than 2.

[0369] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are n-propylene and n-butylene, wherein the number of n-propylene can be 1 or more than 2, and the number of n-butylene can be 1 or more than 2.

[0370] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are n-propylene and isobutylene, wherein the number of n-propylene can be 1 or more than 2, and the number of isobutylene can be 1 or more than 2.

[0371] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are n-propylene and sec-butylene, wherein the number of n-propylene can be 1 or more than 2, and the number of sec-butylene can be 1 or more than 2.

[0372] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are n-propylene and tert-butylene, wherein the number of n-propylene can be 1 or more than 2, and the number of tert-butylene can be 1 or more than 2.

[0373] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are n-propylene and benzylene, wherein the number of n-propylene can be 1 or more than 2, and the number of benzylene can be 1 or more than 2.

[0374] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are isopropylene and n-butylene, wherein the number of isopropylene can be 1 or more than 2, and the number of n-butylene can be 1 or more than 2.

[0375] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are isopropylene and isobutylene, wherein the number of isopropylene can be 1 or more than 2, and the number of isobutylene can be 1 or more than 2.

[0376] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are isopropylene and sec-butylene, wherein the number of isopropylene can be 1 or more than 2, and the number of sec-butylene can be 1 or more than 2.

[0377] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are isopropylene and tert-butylene groups, wherein the number of isopropylene groups can be 1 or more than 2, and the number of tert-butylene groups can be 1 or more than 2.

[0378] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are isopropylene and benzylene, wherein the number of isopropylene can be 1 or more than 2, and the number of benzylene can be 1 or more than 2.

[0379] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are methylene, ethylene and n-propylene, wherein the number of methylene groups can be 1 or more than 2, the number of ethylene groups can be 1 or more than 2, and the number of n-propylene groups can be 1 or more than 2.

[0380] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are methylene, ethylene and isopropylene, wherein the number of methylene groups can be 1 or more, the number of ethylene groups can be 1 or more, and the number of isopropylene groups can be 1 or more.

[0381] In some embodiments, L b When the number is greater than 1, multiple L bIndependently there are methylene, ethylene, n-propylene and isopropylene, wherein the number of methylene groups can be 1 or more, the number of ethylene groups can be 1 or more, the number of n-propylene groups can be 1 or more, and the number of isopropylene groups can be 1 or more.

[0382] In some embodiments, L b When the number is greater than 1, multiple L b Independently there are methylene, ethylene, n-propylene, isopropylene, and n-butylene, wherein the number of methylene groups can be 1 or more, the number of ethylene groups can be 1 or more, the number of n-propylene groups can be 1 or more, the number of isopropylene groups can be 1 or more, and the number of n-butylene groups can be 1 or more.

[0383] In this application, Q a One, two or more selected from -C(=O)NR-, -NR-, -C(=O)O-, -OC(=O)NR-, -NRC(=O)-, -OC(=O)-, -NRC(=O)O-, -NRC(=O)NR-, NHC-(=NH)NH-, -C(=O)-, -OC(=O)O-, wherein R is selected from hydrogen, optionally substituted C1-C6 straight chain or branched alkyl.

[0384] Furthermore, Q a It is one, two, or more than three of -C(=O)-, -NR-, -C(=O)NR-, -NRC(=O)NR-, -OC(=O)O-, -OC(=O)NR-, -NRC(=O)-, or -OC(=O)-, wherein R is selected from hydrogen or an optionally substituted C1-C6 straight or branched alkyl group.

[0385] Furthermore, Q a It is one or two or more of -C(=O)-, -NR-, -NRC(=O)- and -(C=O)NR-, wherein R is selected from hydrogen or an optionally substituted C1-C6 linear or branched alkyl group.

[0386] Furthermore, Q a It is -C(=O)-, -N(CH3)-, -NHC(=O)-, -C(=O)-NH-, -C(=O)-N(CH3)-.

[0387] In some embodiments, Q a When the number of is 1, it is -C(=O)-.

[0388] In some embodiments, Q aWhen the number of is 2, one is -C(=O)- and the other is -C(=O)NH-.

[0389] In some embodiments, Q a When the number of is 2, one is -C(=O)- and the other is -C(=O)-NR-, and R is a methyl group.

[0390] In some embodiments, Q a When the number of is 2, one is -C(=O)-, the other is -NR-, and R is a methyl group.

[0391] In this application, Q b is selected from one, two or more of the group consisting of absent, -C(=O)NR-, -C(=O)O-, -OC(=O)NR-, -NRC(=O)-, -OC(=O)-, -NRC(=O)O-, -NRC(=O)NR-, NHC-(=NH)NH-, -C(=O)-, and -OC(=O)O-, wherein R is selected from hydrogen, an optionally substituted C1-C6 straight chain or branched alkyl group.

[0392] Furthermore, Q b is absent, one or two or more of -C(=O)NR-, -OC(=O)NR-, -NRC(=O)-, -NRC(=O)O-, or -NRC(=O)NR-, and R is selected from hydrogen, an optionally substituted C1-C6 linear or branched alkyl group.

[0393] Furthermore, Q b is absent, one, two, or more than one of -C(=O)-, -NRC(=O)-, -NRC(=O)NR-, or -C(=O)NR-, and R is hydrogen or methyl.

[0394] In some embodiments, Q b In some embodiments, Q b is -C(=O)-. In some embodiments, Q b is -NRC(=O)-, R is hydrogen. In some embodiments, Q b is -NRC(=O)-, R is methyl. In some embodiments, Q b is -NRC(=O)NR-, R is hydrogen. In some embodiments, Q b is -NRC(=O)NR-, R is methyl. In some embodiments, Q b is -C(=O)NR-, R is hydrogen. In some embodiments, Q b is -C(=O)NR-, R is methyl. In the present application, X is selected from:

[0395] The right wavy line in X connects to D;

[0396] R4 is selected from hydrogen, C1-C6 straight or branched alkyl, -(CH2CH2O) m R2, -CH2CH2NR2R3, -CH2CH2SO2R2, R2, R3 are independently selected from hydrogen, C1-C6 straight chain or branched alkyl, and m is an integer greater than 1.

[0397] Furthermore, X is

[0398] R4 is hydrogen, methyl, -CH2CH2OCH2CH2OH, -CH2CH2NMe2, or -CH2CH2SO2Me.

[0399] In some embodiments, X is R4 is hydrogen.

[0400] In some embodiments, X is R4 is methyl.

[0401] In some embodiments, X is R4 is -CH2CH2OCH2CH2OH.

[0402] In some embodiments, X is R4 is -CH2CH2NMe2.

[0403] In some embodiments, X is R4 is -CH2CH2SO2Me.

[0404] In some embodiments, X is R4 is hydrogen.

[0405] In some embodiments, X is R4 is methyl.

[0406] In some embodiments, X is R4 is -CH2CH2OCH2CH2OH.

[0407] In some embodiments, X is R4 is -CH2CH2NMe2.

[0408] In some embodiments, X is R4 is -CH2CH2SO2Me.

[0409] In some embodiments, X is R4 is hydrogen.

[0410] In some embodiments, X is R4 is methyl.

[0411] In some embodiments, X is R4 is -CH2CH2OCH2CH2OH.

[0412] In some embodiments, X is R4 is -CH2CH2NMe2.

[0413] In some embodiments, X is R4 is -CH2CH2SO2Me.

[0414] In the present application, the sulfonic acid inner salt is

[0415] A wavy line connects Y, Y' or Y";

[0416] wherein c, d, e, f, g are all integers of 1, 2, 3, 4, 5 or 6; R6, R7, R 11 、R 12 、R 13 、R 14 are independently selected from absence, hydrogen, and C1-C6 linear or branched alkyl.

[0417] Furthermore, the sulfonic acid inner salt is

[0418] Among them, R6, R7, R 11 、R 12 、R 13 and R 14 are selected from absence, methyl or ethyl, c, d, e, f, g are 1, 2 or 3 respectively.

[0419] Furthermore, the sulfonic acid inner salt is one of the following structures:

[0420] In some embodiments, the sulfonic acid inner salt is

[0421] In some embodiments, the sulfonic acid inner salt is

[0422] In some embodiments, the sulfonic acid inner salt is

[0423] In some embodiments, the sulfonic acid inner salt is

[0424] In this application, the phosphate inner salt is

[0425] Wherein, a and b are integers of 1, 2, 3, 4, 5 or 6; R8, R9 and R 10 are independently selected from hydrogen, C1-C6 straight chain or branched alkyl.

[0426] Furthermore, the phosphate inner salt is

[0427] Among them, R8, R9, R 10 are methyl or ethyl respectively; a is 1 or 2 or 3; b is 1 or 2 or 3.

[0428] Furthermore, the phosphate inner salt is

[0429] Among them, R8, R9, R 10 All are methyl; a is 2; b is 1.

[0430] In the present application, the D includes cytotoxins or immune agonists.

[0431] In the present application, the cytotoxin is selected from one or two of tubulin inhibitors, DNA synthesis inhibitors, topoisomerase inhibitors or RNA polymerase 2 inhibitors.

[0432] In the present application, the cytotoxin is selected from auristatins, maytansine alkaloids DMs, calicheamicin, duocarmycin, pyrrolobenzodiazepines One or more of camptothecin derivatives, camptothecin derivatives and α-amanitin.

[0433] In the present application, the immune agonist is selected from TLR7 / 8 agonist or STING agonist.

[0434] In the present application, i is an integer of 2-8, for example, it can be 2, 3, 4, 5, 6, 7 or 8.

[0435] In the present application, h is an integer of 1-4, for example, it can be 1, 2, 3 or 4.

[0436] In the present application, o, p, and q are integers of 0-3, for example, 0, 1, 2, or 3.

[0437] In this application, L a It is one, two or more of C1-C6 straight or branched alkylene groups, or polyethylene glycol groups, wherein the polyethylene glycol group is -(CH2CH2O) n -, -O(CH2CH2O) n -、-(CH2CH2O) n CH2-, -(CH2CH2O)n CH2CH2-, -CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n CH2-, n is an integer greater than or equal to 1; Q a It is one, two, or more than three of -C(=O)-, -NR(C=O)-, and -(C=O)NR-, wherein R is selected from hydrogen, a C1-C6 linear or branched alkyl group.

[0438] Furthermore, L a is ethylene, linear propylene, linear butylene, linear pentylene, or -(CH2CH2O) n One, two or more of CH2CH2-, wherein n is 2, 3, 4, 5, 6, 7 or 8; Q a It is one, two, or more than three of -C(=O)-, -NR(C=O)-, and -(C=O)NR-, wherein R is methyl and / or hydrogen.

[0439] Furthermore, L a is ethylene and / or -(CH2CH2O)4CH2CH2- or pentylene; Q a is -C(=O)-, -NR-, -NH(C=O)- and / or -C(=O)NH-.

[0440] In some embodiments, L a The number of is 2, one of which is ethylene and the other is -(CH2CH2O)4CH2CH2-. a The number of them is 2, one of which is -C(=O)- and the other is -C(=O)NH-.

[0441] In some embodiments, L a The number is 1, which is pentylene, Q a The number of is 1, which is -C(=O)-.

[0442] In some embodiments, L a The number is 1, which is -(CH2CH2O)4CH2CH2-, Q a The number of them is 2, one is -C(=O)- and the other is -C(=O)NH-.

[0443] In some embodiments, L a The number is 1, which is -(CH2CH2O)4CH2CH2-, Q a The number of them is 2, one is -C(=O)- and the other is -C(=O)N(CH3)-.

[0444] In some embodiments, L a The number of is 2, which are ethylene and pentylene, Q a The number of them is 2, one is -C(=O)- and the other is -N(CH3)-.

[0445] In this application, L b is selected from a C1-C6 straight chain or branched chain alkylene group, a C2-C6 straight chain or branched chain alkenylene group, a C2-C6 straight chain or branched chain alkynylene group, an arylene group having 6 to 10 carbon atoms in one or more rings, or a polyethylene glycol group,

[0446] The polyethylene glycol group is -(CH2CH2O) n -, -O(CH2CH2O) n -、-(CH2CH2O) n CH2-, -(CH2CH2O) n CH2CH2-, -CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n One or two or more of CH2-, n is an integer greater than or equal to 1, and the alkylene, alkenylene, and alkynylene groups are optionally substituted with substituents;

[0447] Q b is selected from one, two or more of absent, -C(=O)NR-, -C(=O)O-, -OC(=O)NR-, -NRC(=O)-, -OC(=O)-, -NRC(=O)O-, -NRS(=O)2-, -NRC(=O)NR-, -C(=O)-, -OC(=O)O-, and R is selected from hydrogen, optionally substituted C1-C6 straight chain or branched alkyl.

[0448] Furthermore, L b is a C1-C6 straight or branched chain alkylene, or a C6-C 10 One or two or more than three arylene groups; Q b It is absent, one or two or more of -C(=O)NR-, -C(=O)-, -OC(=O)NR-, -NRC(=O)-, -NRC(=O)O-, or -NRC(=O)NR-, and R is selected from hydrogen, a C1-C6 straight chain or branched alkyl group.

[0449] Furthermore, L b It is one, two or more of the following: methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, sec-butylene, tert-butylene or benzylene;

[0450] Q bis absent, one, two, or more than one of -C(=O)-, -NRC(=O)-, -NRC(=O)NR-, or -C(=O)NR-, and R is hydrogen or methyl.

[0451] In some embodiments, in Y, L b When the number of is two, one is a methylene group and the other is an isopropylene group; b Does not exist.

[0452] In some embodiments, in Y, L b When the number of is three, they are methylene, isopropylene and ethylene respectively; Q b The number of is 1, which is -C(=O)NR-, and R is hydrogen.

[0453] In some embodiments, in Y, L b When the number of is two, one is ethylene and the other is isopropylene; b The number of is 1, which is -C(=O)NR-, and R is hydrogen.

[0454] In some embodiments, in Y, L b When the number of is two, one is n-propylene and the other is isopropylene; b The number of is 1, which is -NRC(=O)NR-, and R is hydrogen.

[0455] In some embodiments, in Y, L b When the number is 1, it is a benzyl group, Q b Does not exist.

[0456] In some embodiments, in Y, L b When the number of is 2, they are ethylene and isopropylene respectively; Q b The number of is 1, which is -C(=O)-.

[0457] In the present application, the substituent is selected from deuterium, tritium, a halogen atom, an amino group, a hydroxyl group, a cyano group, a nitro group, an optionally substituted alkyl group, an alkenyl group, an alkynyl group, an optionally substituted heterocyclic group, an acyl group, an optionally substituted amino group, an optionally substituted carbamoyl group, an optionally substituted thiocarbamoyl group, an optionally substituted sulfamoyl group, an optionally substituted hydroxyl group, an optionally substituted sulfanyl group (SH) and an optionally substituted silyl group.

[0458] In the present application, the compound is selected from the following structures:

[0459] The present application also provides a compound represented by formula (2), or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein the structure of formula (2) is represented as follows:

[0460] Ab comprises an antibody or an antigen-binding fragment thereof; M' is a linker; Sp is -L a -Q a -; A is selected from the group consisting of non-existent, hydrogen, and alkyl; T represents a linear or branched (C1-C8) trivalent alkyl group; Y, Y', and Y" are respectively -L b -Q b -;L a , L b Q is independently selected from absence, alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene, heteroarylene or polyethylene glycol group, wherein the alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene, heteroarylene or polyethylene glycol group is optionally substituted with a substituent; a , Q b are respectively selected from absence, -C(=O)NR-, -C(=O)O-, -OC(=O)NR-, -S(=O)2NR-, -NRC(=O)-, -OC(=O)-, -NRC(=O)O-, -NRS(=O)2-, -NRC(=O)NR-, -NHC-(=NH)NH-, -C(=O)-, -OC(=O)O-, -O-, -NR-, -S-, S(=O)2-, S(=O)-, or -Se -, wherein R is selected from hydrogen or an optionally substituted alkyl group; Z is selected from absence, hydrogen, an optionally substituted alkyl group, or a sulfonate inner salt; Z' and Z" are selected from absence, hydrogen, an optionally substituted alkyl group, a sulfonate inner salt, or a phosphate inner salt; at least one of Z, Z', and Z" is a sulfonate inner salt or a phosphate inner salt; X comprises a self-destruct unit or is absent; D comprises a drug molecule; i is an integer greater than or equal to 1, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, etc.

[0461] h is an integer from 1 to 6, for example, 1, 2, 3, 4, 5, 6, etc. o, p, and q are integers from 0 to 5, for example, 0, 1, 2, 3, 4, 5, etc. j is an integer greater than or equal to 1, for example, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, etc.

[0462] When h is greater than 1, multiple Sp can have different structures or the same structure, that is, multiple L a It can be of different structures or the same structure; multiple Q aIt can be of different structures or the same structure. a and Q a The number can be the same or different.

[0463] When i is greater than 1, multiple A's may be different or the same, multiple T's may be different or the same, multiple Y's may be different or the same, multiple o's may be different or the same, and multiple Z's may be different or the same.

[0464] When p is greater than 1, the multiple Y's may have different structures or the same structure.

[0465] When j is greater than 1, the parts in the brackets may have the same structure or different structures.

[0466] In this application, the formula (2) is formula (2-1):

[0467] wherein Ab, M', Sp, A, T, Y, h, o, i, j, X, D, and Z are as defined in the aforementioned formula (1).

[0468] At least one of the i Zs is a sulfonate.

[0469] Furthermore, the formula (2-1) is formula (2-1-1):

[0470] Among them, AA is the residue of amino acid,

[0471] w is an integer greater than or equal to 1, for example, 1, 2, 3, 4, 5, 6, etc. When w is greater than 1, the multiple AAs may be residues of the same amino acid or residues of different amino acids.

[0472] In this application, the formula (2) is formula (2-2):

[0473] wherein Ab, M', Sp, A, T, Y, Z, o, p, i, h, j, X, Y', and D are as defined in the aforementioned formula (1); and Z' is a sulfonic acid inner salt or a phosphate inner salt.

[0474] Furthermore, the formula (2-2) is the formula (2-2-1):

[0475] Wherein, AA is an amino acid residue, and w is an integer greater than or equal to 1, for example, 1, 2, 3, 4, 5, 6, etc. When w is greater than 1, multiple AAs can be residues of the same amino acid or residues of different amino acids.

[0476] In this application, the formula (2) is formula (2-3):

[0477] wherein Ab, M', Sp, A, Y, T, Z, o, i, h, j, q, X, Y", and D are as defined in the aforementioned formula (1); and Z" is a sulfonic acid inner salt or a phosphate inner salt.

[0478] Furthermore, the formula (2-3) is formula (2-3-1):

[0479] Wherein, AA is an amino acid residue, and w is an integer greater than or equal to 1, for example, 1, 2, 3, 4, 5, 6, etc. When w is greater than 1, multiple AAs can be residues of the same amino acid or residues of different amino acids.

[0480] In this article, all the definitions described in formula (1) apply to formula (2), all the definitions described in formula (1-1) apply to formula (2-1), all the definitions described in formula (1-1-1) apply to formula (2-1-1), all the definitions described in formula (1-2) apply to formula (2-2), all the definitions described in formula (1-2-1) apply to formula (2-2-1), all the definitions described in formula (1-3) apply to formula (2-3), and all the definitions described in formula (1-3-1) apply to formula (2-3-1). Therefore, for formula (2), formula (2-1), formula (2-1-1), formula (2-2), formula (2-2-1), formula (2-3), and formula (2-3-1), Sp, A, Y, T, Z, o, i, h, j, q, X, Y′, Z′, Y″, Z″, and D in formula (1) can refer to the description in formula (1) and are not further limited here.

[0481] In this application, M' is selected from one of the following structures:

[0482] *Connect Ab.

[0483] Furthermore, M' is selected from one of the following structures:

[0484] In some embodiments, M' is

[0485] In the present application, the compound is selected from the following structures:

[0486] In the present application, the antibody is selected from the group consisting of a murine antibody, a chimeric antibody, a humanized antibody, and a fully human antibody. In the present application, the antibody comprises a monoclonal antibody. In the present application, the antibody comprises a bispecific antibody. In the present application, the antigen-binding fragment is selected from the group consisting of a Fab, Fab', Fv fragment, F(ab')2, F(ab)2, scFv, di-scFv, and VHH.

[0487] In the present application, the Ab is selected from the following group: anti-claudin 18.2 antibody, anti-human folate receptor antibody, anti-HER2 antibody, anti-HER3 antibody, anti-TROP2 antibody, anti-B7-H3 antibody, anti-GPC20 antibody, anti-CDH6 antibody, anti-EGFR antibody, anti-EGFRvIII antibody, anti-AXL antibody, anti-Nectin-4 antibody, anti-Tissue factor antibody, anti-TIM-1 antibody, anti-PSMA antibody, anti-EpCAM antibody, anti-MUC1 antibody, anti-STEAP1 antibody, anti-GPNMB antibody, anti-FGF2 antibody, anti-FOLR1 antibody, anti-c-MET antibody, anti-GFR antibody, anti-AGS-16, anti-Guanylyl cyclase C antibody, anti-Mesothelin antibody, anti-SLC44A4 antibody, anti-EphA2 antibody, anti-AGS-5 antibody, anti-GPC-3 antibody, anti-c-KIT antibody, anti-ROR1 antibody, anti-PD-L1 antibody, anti-CD27L Antibodies, anti-5T4 antibody, anti-Mucin 16 antibody, anti-NaPi2b antibody, anti-STEAP antibody, anti-SLITRK6 antibody, anti-ETBR antibody, anti-BCMA antibody, anti-CEACAM5 antibody, anti-SC-16 antibody, anti-SLC39A6 antibody, anti-Delta-like protein3 antibody, anti-Claudin18.2 antibody, anti-CD19 antibody, anti-CD20 antibody, anti-CD22 antibody, anti-CD30 antibody, anti-CD33 antibody, anti-CD37 antibody, anti-CD45 antibody, anti-CD56 antibody, anti-CD66e antibody, anti-CD70 antibody, anti-CD73 antibody, anti-CD74 antibody, anti-CD79b antibody, anti-CD138 antibody, anti-CD147 antibody, anti-CD166 antibody, anti-CD223 antibody, anti-MUC16 antibody, anti-MSLN antibody, anti-ENPP3 antibody, anti-SLTRK6 antibody, anti-FGFR antibody, anti-LIV-1 antibody, anti-Lewis antibody Y antibody, anti-av-integrin antibody, anti-ASCT2 antibody, anti-C4.4a antibody, anti-CA-IX antibody, anti-CD324 antibody, anti-CD352 antibody, anti-CD44v6 antibody, anti-CD48a antibody, anti-CLL-1 antibody, anti-Cripto antibody, anti-CS1 antibody, anti-DPEP3 antibody, anti-Ephrin-A2 antibody, anti-Ephrin-A4 antibody, anti-ETBR antibody, anti-FGFR2 antibody, anti-FGFR3 antibody, anti-FLT3 antibody, anti-GD3 antibody, anti-GloboH antibody, anti-GPC3 antibody, anti-LAMP-1 antibody, anti-LRRC15 antibody, anti-Ly6E antibody, anti-MFI2 antibody, anti-NOTCH3 antibody, anti-p-cadherin antibody, anti-PRLR antibody and anti-RNF43 antibody, and antigen-binding fragments of the above antibodies.

[0488] In the present application, the heavy chain HCDR1, HCDR2 and HCDR3 and the light chain LCDR1, LCDR2 and LCDR3 of Ab comprise the heavy chain HCDR1, HCDR2 and HCDR3 and the light chain LCDR1, LCDR2 and LCDR3 of the antibody, respectively.

[0489] In the present application, the heavy chain variable region VH and light chain variable region VL of the Ab comprise the heavy chain variable region VH and light chain variable region VL of an antibody, respectively. In the present application, the heavy chain and light chain of the Ab comprise the heavy chain and light chain of an antibody, respectively. In the present application, the Ab is an anti-claudin 18.2 antibody or an antigen-binding fragment thereof. In the present application, wherein the Ab is an anti-human folate receptor antibody or an antigen-binding fragment thereof.

[0490] The present application also provides a pharmaceutical composition, which includes the aforementioned compound, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate.

[0491] The present application also provides the use of the aforementioned compound, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate in the preparation of a drug for treating and / or preventing tumors.

[0492] In the present application, the tumor is selected from tumors associated with the expression of the following group of targets: claudin 18.2, human folate receptor, HER2, HER3, TROP2, B7-H3, GPC20, CDH6, EGFR, EGFRvⅢ, AXL, Nectin-4, Tissue factor, TIM-1, PSMA, EpCAM, MUC1, STEAP1, GPNMB, FGF2, FOLR1, c-MET, GFR, AGS-16, Guanylyl cyclase C, Mesothelin, SLC44A4, EphA2, AGS-5, GPC-3, c-KIT, ROR1, PD-L1, CD27L, 5T4, Mucin 16, NaPi2b, STEAP, SLITRK6, ETBR, BCMA, CEACAM5, SC-16, SLC39A6, Delta-like protein3, Claudin18.2, CD19, CD20, CD22, CD30, CD33, CD37, CD45, CD56, CD66e, CD70, CD73, CD74, CD79b, CD138, CD147, CD166, CD223, MUC16, MSLN, ENPP3, SLTRK6, FGFR, LIV-1, Lewis Y, av-integrin, ASCT2, C4.4a, CA-IX, CD324, CD352, CD44v6, CD48a, CLL-1, Cripto, CS1, DPEP3, Ephrin-A2, Ephrin-A4, ETBR, FGFR2, FGFR3, FLT3, GD3, Globo H, GPC3, LAMP-1, LRRC15, Ly6E, MFI2, NOTCH3, p-cadherin, PRLR, and RNF43.

[0493] In the present application, tumors associated with the expression of the target include tumors with high expression of the target and / or target-positive tumors.

[0494] In the present application, the tumor is selected from solid tumors, blood tumors and metastatic, refractory or recurrent lesions of cancer.

[0495] In the present application, the tumor is selected from the following group: esophageal cancer, gastrointestinal cancer, pancreatic cancer, thyroid cancer, colorectal cancer, renal cancer, lung cancer (e.g., non-small cell lung cancer), liver cancer, gastric cancer, gastric adenocarcinoma, gastroesophageal junction (GEJ) adenocarcinoma, head and neck cancer, bladder cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, germ cell cancer, bone cancer, skin cancer, thymic cancer, bile duct cancer, gallbladder cancer, melanoma, mesothelioma, lymphoma, myeloma (e.g., multiple myeloma), sarcoma, glioblastoma and leukemia.

[0496] Example

[0497] Unless otherwise specified, the experimental methods used in the following examples are all conventional methods.

[0498] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0499] In the following examples, Fmoc-L-glutamic acid-5-tert-butyl ester, compound 12-1, HATU (Chinese name is 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate), DIEA (Chinese name is N,N-diisopropylethylamine), di(p-nitrobenzene) carbonate, exitecan mesylate, Mal-amido-PEG4-acid (Chinese name is maleimide-amide-PEG4-propionic acid), DCM (Chinese name is dichloromethane), TFA ( Chinese name is trifluoroacetic acid), 6-maleimidocaproic acid, Fmoc-L-valyl-L-alanine, di(p-nitrophenyl) carbonate, Fmoc-L-glutamic acid-5-tert-butyl ester, 4-aminobenzyl alcohol, Fmoc-L-valine, MMAE (Chinese name is monomethyl auristatin E), HOBT (Chinese name is 1-hydroxybenzotriazole), methyl tert-ether, compound 65-1, BH3-Me2S, DMAP (Chinese name is 4-dimethylaminopyridine), (BOC)2O, EA (Chinese name called ethyl acrylate), TBAF (Chinese name is tetra-n-butylammonium fluoride), EEDQ (Chinese name is 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline), compound 71-1, 6-(maleimido)hexanoic acid succinimide ester, compound 72-1, HOSu (Chinese name is N-hydroxysuccinimide), EDCI (Chinese name is carbodiimide), Fmoc-glycine, nitrophenyl chloroformate, compound 73-1, BCN-PEG4-acid (Chinese name is re l-1-((1R,8S,9s)-bicyclo[6.1.0]non-4-yn-9-yl)-3-oxo-2,7,10,13,16-pentaoxa-4-azanonadecan-19-oic acid), Fmoc-NMe-PEG4-C2-Acid, N,N-diisopropylethylamine, 4-methylsulfonylbenzoic acid, DIPEA (Chinese name is N,N-diisopropylethylamine), exo-3,6-epoxy-1,2,3,6-tetrahydrophthalic anhydride, and N-Boc-N-methylethylenediamine are all commercially available.

[0500] Example 1 (Preparation of LD-12)

[0501] The synthetic route of LD-12 is as follows:

[0502] Synthesis of 12-2

[0503] Fmoc-5-tert-butyl L-glutamate (600 mg, 1.41 mmol) and compound 12-1 (414 mg, 1.41 mmol) were dissolved in 18 mL of anhydrous DMF. HATU (644 mg, 1.69 mmol) and DIEA (365 mg, 2.82 mmol) were added. Stirring was continued at room temperature for 18 hours. LCMS indicated the reaction was complete. Water was added, and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to yield 0.9 g of the crude product.

[0504] Synthesis of 12-3

[0505] Compound 12-2 (300 mg, 0.43 mmol), di(p-nitrobenzene) carbonate (261 mg, 0.86 mmol), and DIEA (166 mg, 1.28 mmol) were dissolved in 4 mL of anhydrous DMF and stirred at room temperature for 18 hours. LCMS indicated the reaction was complete. Water was added, and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification by TLC (MeOH / DCM, 0% to 3%) yielded 0.3 g. MS+Na(+) 889.4.

[0506] Synthesis of 12-4

[0507] Compound 12-3 (350 mg, 0.425 mmol), DIEA (207 mg, 1.61 mmol), and isotecan mesylate (600 mg, 0.69 mmol) were dissolved in 2 mL of anhydrous DMF and reacted at room temperature for 6 hours. Water was then added and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed using a normal phase silica gel column (MeOH / DCM, 0% to 10%) to yield 470 mg. MS(+) was 1162.4.

[0508] Synthesis of 12-5

[0509] Compound 12-4 (100 mg, 0.086 mmol) was dissolved in 2 mL of anhydrous DMF. Piperidine (0.05 mL) was added and stirred at room temperature for 2 hours. After concentration under reduced pressure, the mixture was slurried with tert-methyl ether and filtered to obtain 80 mg of crude product, which was used directly in the next reaction. MS(+) 940.4.

[0510] Synthesis of 12-6

[0511] Compound 12-5 (46 mg, 0.11 mmol) was dissolved in 2 mL of anhydrous DMF, and HATU (48.5 mg, 0.13 mmol) was added. The mixture was stirred at room temperature for five minutes. Mal-amido-PEG4-acid (80 mg, 0.085 mmol) and DIEA (33 mg, 0.26 mmol) were added sequentially. Stirring was continued at room temperature for two hours. LCMS indicated the reaction was complete. Water was added, and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed by TLC (MeOH / DCM, 0% to 3%) to yield 30 mg. MS(+) 1339.2.

[0512] Synthesis of LD-12

[0513] Compound 12-6 (30 mg, 0.022 mmol) was dissolved in 2 mL DCM, 0.3 mL TFA was added at room temperature and stirred for 2 hours. LCMS showed that the reaction was complete. After concentration under reduced pressure, the mixture was prepared by reverse phase (mobile phase containing 0.1% formic acid) to obtain 7.7 mg.

[0514] LCMS: m / z(ES+)(M+H)+=1282.4, Rt=1.385min

[0515] 1H NMR (400MHz, DMSO-d6) δ10.00(s,1H),8.23(d,J=6.8Hz,1H),8.11-8.03(m,3H),7.79(d,J=10.8Hz,1H),7.72(d,J=8.4Hz,1H),7. 60(d,J=8.4Hz,2H),7.38(d,J=8.4Hz,2H),7.33(s,1H),7.00(s,2H),6.55(s,1H),5.46(s,2H),5.36-5.25(m,3H),5.09(s,2H),4 .41-4.32(m,2H),4.23-4.17(m,1H),3.60(t,J=7.2Hz,2H),3.50-3.49(m,16H),3.37-3.33(m,2H),3.16-3.14(m,2H),2.45-2.32 (m,7H),2.27-2.10(m,4H),2.01-1.96(m,1H),1.96-1.81(m,3H),1.74(d,J=6.8Hz,1H),1.31(d,J=7.2Hz,3H),0.91-0.83(m,9H).

[0516] Example 2 (Preparation of LD-27)

[0517] The process flow of LD-27 is as follows:

[0518] Synthesis of 27-1

[0519] Compound 12-5 (127.34 mg, 0.306 mmol; see Example 1 for the synthesis of compound 12-5) and 6-maleimidocaproic acid (80 mg, 0.085 mmol) were dissolved in 4 mL of anhydrous DMF. HATU (49 mg, 0.128 mmol) and DIEA (22 mg, 0.170 mmol) were added and stirred at room temperature for 3 hours. LCMS indicated the reaction was complete. Water was added, and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to yield 80 mg of crude product. MS(+) = 1133.4.

[0520] Synthesis of LD-27

[0521] Compound 27-1 (70 mg, 0.062 mmol) was dissolved in 5 mL DCM, 0.8 mL TFA was added at room temperature and stirred for 2 hours. LCMS showed that the reaction was complete. After concentration under reduced pressure, the mixture was prepared by reverse phase (mobile phase containing 0.1% formic acid) to obtain 22.8 mg.

[0522] LCMS: m / z(ES+)(M+H)+=1077.4, Rt=1.454min

[0523] 1H NMR (400MHz, DMSO-d6) δ12.08(s,1H),9.97(s,1H),8.21(d,J=6.8Hz,1H),8.06-7.98(m,2 H),7.77(d,J=10.8Hz,1H),7.65-7.57(m,3H),7.44-7.26(m,3H),6.99(s,2H),6.51(s,1H ),5.45(s,2H),5.28(d,J=4.4Hz,3H),5.08(s,2H),4.42-4.15(m,3H),3.26-3.08(m,2H), 2.37(s,3H),2.29-1.63(m,12H),1.51-1.42(m,4H),1.36-1.12(m,6H),0.89-0.81(m,9H).

[0524] Example 3 (Preparation of LD-31)

[0525] The synthetic route of LD-31 is as follows:

[0526] Synthesis of 31-2

[0527] Fmoc-L-valyl-L-alanine (220 mg, 0.54 mmol) was dissolved in 5 mL of anhydrous DMF and HATU (244.5 mg, 0.643 mmol) and DIEA (138.5 mg, 1.07 mmol) were added. After stirring at room temperature for 15 minutes, compound 31-1 (119.67 mg, 0.536 mmol, provided by Shanghai Haoyuan Pharmaceutical Co., Ltd., synthesis method reference WO2022058548A1) was added to the reaction solution. After stirring at room temperature for 2 hours, LCMS monitored the reaction to be complete. The crude product was concentrated and purified using a normal phase silica gel column (MeOH / DCM, 0% to 10%). 320 mg was obtained, MS+Na(+) = 638.4.

[0528] Synthesis of 31-3

[0529] Compound 31-2 (320 mg, 0.520 mmol) was dissolved in 5 mL of DCM. Di(p-nitrobenzene) carbonate (474.30 mg, 1.559 mmol) and DIEA (0.344 mL, 2.079 mmol) were added sequentially and stirred at room temperature for 2 hours. LCMS monitored the reaction for completion. Saturated brine was added and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed on a normal phase silica gel column (MeOH / DCM, 0% to 10%). The yield was 340 mg, MS+Na(+) = 803.2.

[0530] Synthesis of 31-4

[0531] Compound 31-3 (340 mg, 0.462 mmol) was dissolved in 2 mL of anhydrous DMF. Isotecan mesylate (245.61 mg, 0.666 mmol), DIEA (119.43 mg, 0.924 mmol), and DMAP (5.65 mg, 122.17 mmol) were added sequentially. After stirring at room temperature for 1 hour, the reaction was complete as monitored by LCMS. Saturated brine was added to precipitate a crude solid. The crude product was slurried with tert-methyl ether to yield 157 mg of compound 31-4, MS(+) = 1078.2.

[0532] Synthesis of 31-5

[0533] Compound 31-4 (440 mg, 0.408 mmol) was dissolved in 4 mL of anhydrous DMF, and piperidine (69.56 mg) was added to the reaction solution. The mixture was stirred at room temperature for one hour. The solution was concentrated to obtain 292 mg of crude product, which was used directly in the next reaction. MS(+)=855.2.

[0534] Synthesis of 31-6

[0535] Compound 31-5 (142.23 mg, 0.342 mmol) and Mal-amido-PEG4-acid (292 mg, 0.342 mmol) were dissolved in 4 mL of anhydrous DMF. DIEA (88.28 mg, 0.683 mmol) and HATU (155.84 mg, 0.410 mmol) were added. The mixture was stirred at room temperature for 2 hours. LCMS indicated the reaction was complete. Saturated brine was added to precipitate a crude solid. The crude product was slurried with tert-methyl ether to yield 350 mg of crude product, MS(+) = 1254.4.

[0536] Synthesis of 31-7

[0537] Compound 31-6 (90 mg crude product) was dissolved in 2.5 mL of anhydrous DCM, TFA (0.5 mL) was added to the reaction solution, and stirred at room temperature for 1 hour. The solution was concentrated to obtain 80 mg crude product, which was used directly in the next reaction. MS (+) = 1198.4.

[0538] Synthesis of LD-31

[0539] Compound 31-7 (76.62 mg, 0.064 mmol) was dissolved in 2 mL of anhydrous DMF. DIEA (16.54 mg, 0.128 mmol) and HATU (29.20 mg, 0.077 mmol) were added and stirred at room temperature for 15 minutes. Sulfonic acid quaternary ammonium salt A (14.81 mg, 0.070 mmol, CAS number: 78276-19-4, provided by Shanghai Haoyuan Pharmaceutical Co., Ltd., synthesis method reference WO2011146595A2) was added and stirred at room temperature for 2 hours. LCMS indicated the reaction was complete. Water was added and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification by reverse phase preparative (formic acid) afforded LD-31 (P1: 12.5 mg, P2: 11.3 mg) as a white solid.

[0540] LCMS: m / z(ES+)(M+H)+=1390.6, Rt=1.242min(P1), 1.279min(P2).

[0541] Example 4 (Preparation of LD-37)

[0542] The synthetic route of LD-37 is as follows:

[0543] Synthesis of LD-37

[0544] Compound LD-12 (49 mg, 0.038 mmol, see Example 1 for the synthesis of LD-12) was dissolved in 2 mL of anhydrous DMF. DIEA (9.88 mg, 0.076 mmol) and HATU (17.43 mg, 0.046 mmol) were added and stirred at room temperature for 15 minutes. Sulfonic acid quaternary ammonium salt A (8.04 mg, 0.038 mmol, CAS No. 78276-19-4, provided by Shanghai Haoyuan Pharmaceutical Co., Ltd., synthesis method reference WO2011146595A2) was added and stirred at room temperature for 2 hours. LCMS indicated the reaction was complete. Water was added and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed by reverse phase preparative (mobile phase containing 0.1% formic acid) to obtain LD-37 (7.35 mg, 13.04% yield) as a white solid.

[0545] LCMS: m / z(ES+)(M+H)+=1475.6, Rt=1.344min

[0546] 1H NMR (400MHz, DMSO) δ10.03(s,1H),8.24(d,J=6.4Hz,1H),8.16-8.07(m,3H),8.03(t,J=6.0Hz,1H),7.80(d,J=10.8Hz,1H),7.71(d,J=9.2Hz,1H), 7.62(d,J=8.0Hz,2H),7.38(d,J=8.4Hz,2H),7.33(s,1H),7.02(s,2H),6 .53(s,1H),5.47(s,2H),5.31(s,3H),5.10(s,2H),4.41(d,J=6.8Hz,1H), 4.31(s,1H),4.24-4.20(m,1H),3.61(t,J=7.2Hz,5H),3.46-3.51(m,16H ),3.16(dd,J=11.2,5.6Hz,4H),3.05(s,6H),2.69(s,3H),2.40(s,4H),2. 35(s,6H),2.18(s,4H),2.01(s,2H),1.90(d,J=9.2Hz,2H),1.79-1.69(m ,1H),1.32(d,J=7.2Hz,3H),0.89(d,J=6.8Hz,6H),0.84(d,J=6.8Hz,3H).

[0547] Example 5: (Preparation of LD-38)

[0548] The synthetic route of LD-38 is as follows:

[0549] Synthesis of 38-1

[0550] Fmoc-5-tert-butyl L-glutamate (144.7 mg, 1.175 mmol) and 4-aminobenzyl alcohol (500 mg, 1.175 mmol) were dissolved in 3 mL of anhydrous DMF. DIEA (303.7 mg, 2.350 mmol) and HATU (536.19 mg, 1.410 mmol) were added and stirred at room temperature for 16 hours. LCMS indicated the reaction was complete. Water was added and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed using a normal phase silica gel column (EA / PE, 0% to 45%) to yield 530 mg. MS(+) = 475.2.

[0551] Synthesis of 38-2

[0552] Compound 38-1 (530 mg, 0.999 mmol) was dissolved in 3 mL of anhydrous DMF, piperidine (0.2 mL) was added to the reaction solution, and the mixture was stirred at room temperature for one hour. The solution was concentrated to obtain 600 mg of crude product, which was used directly in the next reaction. MS(+)=309.2.

[0553] Synthesis of 38-3

[0554] Compound 38-2 (600 mg crude product) and Fmoc-L-valine (500 mg, 1.175 mmol) were dissolved in 3 mL of anhydrous DMF. DIEA (334.13 mg, 2.585 mmol) and HATU (589.82 mg, 1.551 mmol) were added and stirred at room temperature for 16 hours. LCMS showed the reaction was complete. Water was added and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed using a normal phase silica gel column (EA / PE, 40% to 60%) to obtain 450 mg of the product. MS(+) = 630.4.

[0555] Synthesis of 38-4

[0556] Compound 38-3 (450 mg, 0.715 mmol) was dissolved in 1 mL of anhydrous DCM and 4 mL of anhydrous DMF. Di(p-nitrobenzene) carbonate (869.5 mg, 2.858 mmol) and 0.354 mL of DIEA were added sequentially. After reacting at room temperature for 12 hours, the reaction mixture was added with water and extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed using a normal phase silica gel column (EA / PE, 30% to 70%) to yield 200 mg. MS(+) = 543.2.

[0557] Synthesis of 38-5

[0558] Compound 38-4 (100 mg, 0.126 mmol) and isotecan mesylate (54.78 mg, 0.126 mmol) were dissolved in 2 mL of anhydrous DMF. DMAP (0.15 mg, 0.0001 mmol) and DIEA (32.52 mg, 0.252 mmol) were added sequentially. After reacting at room temperature for 1 hour, the reaction mixture was added with water and extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed using a normal phase silica gel column (EA / PE, 30% to 70%) to yield 100 mg. MS(+) = 1091.4.

[0559] Synthesis of 38-6

[0560] Compound 38-5 (100 mg, 0.073 mmol) was dissolved in 3 mL of anhydrous DMF. Piperidine (0.3 mL) was added to the reaction mixture and stirred at room temperature for one hour. The mixture was concentrated to obtain 110 mg of crude product, which was used directly in the next reaction. MS(+) = 870.8.

[0561] Synthesis of 38-7

[0562] Compound 38-6 (110 mg crude) and Mal-amido-PEG4-acid (30.19 mg, 0.073 mmol) were dissolved in 2 mL of anhydrous DMF. DIEA (18.74 mg, 0.145 mmol) and HATU (33.08 mg, 0.087 mmol) were added and stirred at room temperature for 2 hours. LCMS indicated the reaction was complete. Water was added and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed using a normal phase silica gel column (MeOH / DCM, 5% to 20%) to yield 90 mg. MS(+) = 1267.6.

[0563] Synthesis of 38-8

[0564] Compound 38-7 (90 mg, 0.071 mmol) was dissolved in 3 mL of anhydrous DCM, and 0.3 mL of TFA was added. The mixture was stirred at room temperature for 2 hours. Concentration afforded 80 mg of crude product, which was used directly in the next reaction. MS(+) = 2111.4.

[0565] Synthesis of LD-38

[0566] Compound 38-8 (80 mg, crude) and quaternary ammonium sulfonate salt A (13.89 mg, 0.066 mmol, CAS No. 78276-19-4, provided by Shanghai Haoyuan Pharmaceutical Co., Ltd., synthesis method reference WO2011146595A2) were dissolved in 2 mL of anhydrous DMF. DIEA (17.07 mg, 0.132 mmol) and HATU (30.14 mg, 0.079 mmol) were added. The mixture was stirred at room temperature for 1 hour. LCMS indicated the reaction was complete. Purification was performed by reverse phase preparative chromatography (mobile phase containing 0.1% formic acid) to yield 13.5 mg of a white solid.

[0567] LCMS: m / z(ES+)(M+H)+=1404.6, Rt=1.314min

[0568] 1HNMR (400MHz, DMSO) δ9.89 (s, 1H), 8.18 (d, J = 7.2Hz, 2H), 8.10-7.93 (m, 3H), 7.78 (d,J=10.8Hz,1H),7.63(d,J=8.4Hz,2H),7.37(d,J=8.4Hz,2H),7.32(s,1H),6.99 (s,2H),6.53(s,1H),5.45(s,2H),5.29(d,J=4.Hz,3H),5.08(s,2H),4.32(dd,J=13.2,7.6Hz,1H),4.20-4.14(m,1H),3.59(dd,J=14.8,7. 6Hz,6H),3.40-3.51(m,18H),3.14-3.20(m,4H),3.03(s,7H),2.42-2.30(m,7H),2.20-2.31(m,4H),2.03-1.82(m,8H),0.87-0.91(m,9H).

[0569] Example 6 (Preparation of LD-57)

[0570] The synthetic route of LD-57 is as follows:

[0571] Synthesis of 57-1

[0572] Compound 38-4 (288 mg, 0.363 mmol; see Example 5 for its synthesis) and MMAE (200 mg, 0.279 mmol) were dissolved in 5 mL of anhydrous DMF. Pyridine (1 mL) and HOBT (59 mg, 0.279 mmol) were added sequentially. After reaction at room temperature for 18 hours, the crude product was concentrated to afford the product. Purification was performed using a normal phase silica gel column (MeOH / DCM, 0% to 10%) to afford 320 mg. MS(+) = 1374.2.

[0573] Synthesis of 57-2

[0574] Compound 57-1 (260 mg, 0.189 mmol) was dissolved in 3 mL of anhydrous DMF. Piperidine (0.05 mL) was added to the reaction mixture and stirred at room temperature for one hour. The mixture was concentrated and slurried with tert-methyl ether to obtain 215 mg of crude product, which was used directly in the next reaction. MS(+) = 1152.5.

[0575] Synthesis of 57-3

[0576] Compound 57-2 (215 mg crude product) and 6-maleimidocaproic acid (47.3 mg, 0.224 mmol) were dissolved in 3 mL of anhydrous DMF. DIEA (0.092 mL, 0.559 mmol) and HATU (142 mg, 0.374 mmol) were added and stirred at room temperature for 2 hours. LCMS indicated the reaction was complete. Water was added and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed using a normal phase silica gel column (MeOH / DCM, 0% to 10%) to yield 170 mg. MS(+) = 1345.7.

[0577] Synthesis of 57-4

[0578] Compound 57-3 (150 mg, 0.112 mmol) was dissolved in 3 mL of anhydrous DCM. TFA (0.6 mL) was added to the reaction mixture and stirred at room temperature for 2 hours. The mixture was concentrated to obtain 150 mg of crude product, which was used directly in the next reaction. MS(+) = 1289.5.

[0579] Synthesis of LD-57

[0580] Compound 57-4 (143 mg, 0.111 mmol) and quaternary ammonium sulfonate A (70 mg, 0.333 mmol, CAS No. 78276-19-4, provided by Shanghai Haoyuan Pharmaceutical Co., Ltd., synthesis method reference WO2011146595A2) were dissolved in 2 mL of anhydrous DMF. DIEA (0.092 mL, 0.554 mmol) and HATU (126.5 mg, 0.333 mmol) were added. The mixture was stirred at room temperature for 3 hours. LCMS indicated the reaction was complete. Purification was performed by reverse phase preparative chromatography (mobile phase containing 0.1% formic acid) to yield 30 mg of a white solid.

[0581] LCMS: m / z(ES+)(M+H)+=1481.2, Rt=1.475min

[0582] 1HNMR (400MHz, DMSO) δ9.89 (s, 1H), 8.40-8.03 (m, 3H), 7.92 (t, J = 7.6Hz, 1H),7.63(d,J=6.4Hz,2H),7.34-7.13(m,8H),7.02(s,2H),5.06-5.10(m, 3H),4.85-3.92(m,10H),3.63-3.43(m,9H),3.30-2.97(m,18H),2.89(d,J =14.4Hz,3H),2.37-1.68(m,17H),1.61-1.45(m,6H),1.40-0.70(m,36H).

[0583] Example 7 (Preparation of LD-65)

[0584] The synthetic route of LD-67 is as follows:

[0585] Synthesis of 65-2

[0586] Compound 65-1 (500 mg, 2.791 mmol) was dissolved in 5 mL of methanol and methylamine methanol solution (1.73 g, 55.825 mmol) was added. After reacting at room temperature for 1 hour, the crude product was filtered and washed twice with water and dried under reduced pressure to yield 550 mg. MS(+) 211.2.

[0587] Synthesis of 65-3

[0588] Compound 65-2 (500 mg, 2.379 mmol) was dissolved in 50 mL of anhydrous THF. BH₃-Me₂S (457 mg, 5.947 mmol) (10.0 M in THF) was slowly added dropwise under ice-cooling. After reflux for 5 hours, LCMS indicated the reaction was complete. 4M HCl (12 mL) in methanol was added to the reaction mixture under ice-cooling, and the reaction was continued by heating at 65°C for 8 hours. The mixture was filtered and dried under reduced pressure to yield 350 mg. MS(+)=197.08.

[0589] Synthesis of 65-4

[0590] Compound 65-3 (350 mg crude product) and imidazole (485.77 mg, 7.135 mmol) were dissolved in 5 mL of anhydrous DCM. TBDPS-Cl (735.44 mg, 2.676 mmol) was added at 0°C and stirred for 2 hours. LCMS indicated the reaction was complete. Water was added and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed using a normal phase silica gel column (EA / PE, 10% to 50%) to yield 710 mg. MS(+) 435.20.

[0591] Synthesis of 65-5

[0592] Compound 64-4 (1.5 g, 3.451 mmol) and DMAP (0.21 g, 1.726 mmol) were dissolved in DCM (20 mL). (BOC)2O (1.51 g, 6.903 mmol) was added and stirred for 2 hours. Water was added and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed using a normal phase silica gel column (EA / PE, 10% to 50%) to yield 1.87 g. MS+Na(+) 557.3.

[0593] Synthesis of 65-6

[0594] Compound 65-5 (1.7 g, 3.179 mmol) and Pd / C 5% (0.15 g, 1.410 mmol) were dissolved in EA (30 mL). The reaction mixture was stirred under a hydrogen atmosphere for 16 hours, filtered, and dried to obtain 1.7 g. MS+Na(+) 527.6.

[0595] Synthesis of 65-7

[0596] Compound 65-6 (350 mg, 0.693 mmol) was dissolved in anhydrous THF (4 mL), and TBAF (1.387 mL, 1.387 mmol) was added and stirred at room temperature for 4 hours. The reaction mixture was spin-dried and purified on a normal phase silica gel column (MeOH / DCM, 0% to 10%) to yield 170 mg. MS+Na(+) = 289.2.

[0597] Synthesis of 65-8

[0598] Compound 65-7 (170 mg, 0.638 mmol), Fmoc-L-valyl-L-alanine (392.99 mg, 0.957 mmol), and EEDQ (236.48 mg, 0.957 mmol) were dissolved in 2 mL of DCM and 1 mL of methanol and stirred at room temperature for 16 hours. The reaction mixture was then dried and purified on a normal phase silica gel column (MeOH / DCM, 0% to 10%) to yield 367 mg. MS(+)Na(+)=681.4.

[0599] Synthesis of 65-9

[0600] Compound 65-9 (367 mg, 0.557 mmol), di(p-nitrobenzene) carbonate (224.57 mg, 1.114 mmol), and pyridine (0.090 mL, 1.114 mmol) were dissolved in 5 mL of THF and stirred at room temperature for 16 hours. LCMS indicated the reaction was complete. Water was added, and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed using a normal phase silica gel column (MeOH / DCM, 0% to 10%) to yield 380 mg. MS(+)Na(+)=846.4.

[0601] Synthesis of 65-10

[0602] Compound 65-9 (350 mg, 0.425 mmol), DMAP (5.19 mg, 0.043 mmol), and isotecan mesylate (248.50 mg, 0.468 mmol) were dissolved in 2 mL of anhydrous DMF and reacted at room temperature for 2 hours. Water was then added and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed using a normal phase silica gel column (MeOH / DCM, 0% to 10%) to yield 350 mg. MS(+) was 1120.6.

[0603] Synthesis of 65-11

[0604] Compound 65-10 (350 mg, 0.312 mmol) was dissolved in 2 mL of anhydrous DMF. Pyridine (0.05 mL) was added and stirred at room temperature for 2 hours. After concentration under reduced pressure, 380 mg of the crude product was used directly in the next reaction. MS(+) = 898.41.

[0605] Synthesis of 65-12

[0606] Compound 65-11 (127.34 mg, 0.306 mmol) was dissolved in 3 mL of anhydrous DMF, and HATU (116.28 mg, 0.306 mmol) was added. The mixture was stirred at room temperature for five minutes. Mal-amido-PEG4-acid (250 mg, 0.278 mmol) and DIEA (3 mL) were added sequentially. Stirring was continued at room temperature for two hours. LCMS indicated the reaction was complete. Water was added, and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed using a normal phase silica gel column (MeOH / DCM, 0% to 10%) to yield 185 mg. MS(+) = 1296.6.

[0607] Synthesis of 65-13

[0608] Compound 65-12 (40 mg, 0.031 mmol) was dissolved in 3 mL of DCM, 0.6 mL of TFA was added at room temperature, and the mixture was stirred for 2 hours. LCMS showed that the reaction was complete, and the mixture was concentrated under reduced pressure to yield 37 mg. MS+Na(+)=1218.6.

[0609] Synthesis of LD-65

[0610] Sulfonic acid quaternary ammonium salt B (37 mg, 0.031 mmol, CAS number: 1349808-80-5, provided by Shanghai Haoyuan Pharmaceutical Co., Ltd., synthesis method reference WO2011146595A2) was dissolved in 2 mL of anhydrous DMF. HATU (12.94 mg, 0.034 mmol) was added and stirred at room temperature for five minutes. Compound 65-13 (11.75 mg, 0.046 mmol) and DIEA (0.008 mL, 0.046 mmol) were added sequentially. Stirring was continued at room temperature for 2 hours. LCMS indicated the reaction was complete. Reverse phase preparation (mobile phase containing 10 mM NH4OAc) was used to obtain 5.2 mg of a white solid.

[0611] LCMS: m / z(ES+)(M+H)+=1432.6, Rt=1.285min

[0612] 1H NMR (400MHz, DMSO) δ10.2-9.9(m,1H),8.19(t,J=7.2Hz,1H),8.12-8.00(m,2H),7.90-7.85(m,1H),7. 78(d,J=10.8Hz,1H),7.39-7.30(m,3H),7.00(s,2H),6.52(s,1H),5.44(d,J=4.0Hz,2H),5.29(s,4H) ,5.12(d,J=10.8Hz,2H),4.64-4.70(m,2H),4.39-4.34(m,1H),4.22-4.17(m,1H),3.48-3.47(m,16H) ,3.04-2.86(m,14H),2.39-2.30(m,11H),2.03-1.85(m,10H),1.29-1.30(m,3H),0.897-0.817(m,9H).

[0613] Example 8 (Preparation of LD-66)

[0614] The synthetic route of LD-66 is as follows:

[0615] Synthesis of LD-66

[0616] Quaternary ammonium phosphate (8.93 mg, 0.046 mmol, Cas No.: 853798-56-8, provided by Shanghai Haoyuan Pharmaceutical Co., Ltd., synthesis method reference WO2005114193) was dissolved in 2 mL of anhydrous DMF. DIEA (0.008 mL, 0.046 mmol) and HATU (12.94 mg, 0.034 mmol) were added and stirred at room temperature for 5 minutes. Compound 65-13 (37 mg, 0.031 mmol) was added and stirred for 2 hours. LCMS indicated the reaction was complete. Purification was performed by reverse phase preparative (formic acid) to yield 11.5 mg of a white solid.

[0617] LCMS: m / z(ES+)(M+H)+=1420.6, Rt=1.246min

[0618] 1H NMR (400MHz, DMSO) δ10.11(s,1H),8.23-8.17(m,1H),8.07-7.87(m,2H),7.79-7.69(m,2H),7.45-7.1 3(m,4H),7.00(s,2H),6.51(s,1H),5.47-5.06(m,8H),4.83(dd,J=45.6,17.8Hz,1H),4.59(d,J=7.6Hz ,1H),4.48(d,J=8.4Hz,1H),4.37(d,J=6.8Hz,1H),4.21-4.09(m,4H),3.49-3.48(m,16H),3.19-3.10 (m,12H),2.96(s,5H),2.39-2.32(m,8H),1.99-1.83(m,4H),1.30(d,J=6.8Hz,3H),0.90-0.82(m,9H).

[0619] Example 9 (Preparation of LD-71)

[0620] The synthetic route of LD-71 is as follows:

[0621] Synthesis of 71-2

[0622] Compound 31-1 (240 mg, 1.075 mmol, for its synthesis, see Example 1) and compound 71-1 (267 mg, 0.538 mmol, provided by Shanghai Haoyuan Pharmaceutical Co., Ltd., for its synthesis method, see WO2022058548A1) were dissolved in 3 mL of anhydrous DCM and 1.5 mL of methanol, and EEDQ (266 mg, 1.075 mmol) was added. The mixture was stirred at room temperature for 3 hours, during which time a solid precipitated. LCMS monitored the reaction for completion. Filter the filter cake, wash it three times with DCM, and dry it under reduced pressure to yield 260 mg. MS(+) = 702.4.

[0623] Synthesis of 71-3

[0624] Compound 71-2 (240 mg, 0.342 mmol), di(p-nitrobenzene) carbonate (312.09 mg, 1.026 mmol), and DIEA (0.170 mL, 1.026 mmol) were dissolved in 5 mL of DMF and stirred at room temperature for 16 hours. The reaction was monitored for completion by LCMS. Water was added and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed using a normal phase silica gel column (MeOH / DCM, 5% to 10%) to yield 220 mg. MS(+) = 867.4.

[0625] Synthesis of 71-5

[0626] Under nitrogen at room temperature, triphosgene (136.1 mg, 0.459 mmol) was dissolved in 4 mL of anhydrous DCM. Triethylamine (0.128 mL, 0.917 mol) and 71-4 (172.6 mg, 0.917 mol) were added dropwise under an ice bath. After stirring at room temperature for 3 hours, pyridine (2 mL), SN38 (120 mg, 0.306 mmol), and DMAP (7.47 mg, 0.061 mol) were added. The reaction was stirred at 50°C for 16 hours, and the reaction was complete as monitored by LCMS. The product was concentrated to dryness, slurried with methyl tert-ether, and filtered to yield 127 mg. MS(+) = 607.4.

[0627] Synthesis of 71-6

[0628] Compound 71-5 (140 mg, 0.231 mmol) was dissolved in 3 mL of anhydrous DCM, 0.6 mL of TFA was added at room temperature and stirred for 2 hours. LCMS showed that the reaction was complete. After concentration under reduced pressure, 116.9 mg was obtained. MS (+) = 507.2.

[0629] Synthesis of 71-7

[0630] Compound 71-6 (116.9 mg, 0.231 mmol) and compound 71-3 (200 mg, 0.231 mmol) were dissolved in 2 mL of DMSO and 1 mL of DMF. HOBT (31.17 mg, 0.231 mmol) and DIEA (59.64 mg, 0.461 mmol) were added sequentially. After stirring at 40°C for 3 hours, the reaction mixture was added with saturated brine to obtain the crude product. Purification was performed using a normal phase silica gel column (MeOH, 0% to 10%) to yield 200 mg. MS(+) = 1234.6.

[0631] Synthesis of 71-8

[0632] Dissolve 100 mg of crude compound 71-7 in 2 mL of anhydrous DMF, add pyridine (0.05 mL) at room temperature, and stir for 2 hours. After concentration under reduced pressure, the mixture was slurried with tert-methyl ether and filtered to obtain 100 mg of crude product, which was used directly in the next reaction. MS(+) 1012.6.

[0633] Synthesis of 71-9

[0634] Dissolve 100 mg of crude compound 71-8 and 6-(maleimido)hexanoic acid succinimidyl ester (43.9 mg, 0.142 mmol) in 2 mL of anhydrous DMF, add DIEA, and stir at room temperature for 2 hours. Water is added to the reaction mixture, and the mixture is extracted three times with ethyl acetate. The combined organic phases are washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification is performed on a thin-layer silica gel plate (MeOH / DCM, 0% to 10%) to yield 60 mg. MS(+) = 1205.6.

[0635] Synthesis of 71-10

[0636] Compound 71-9 (60 mg, 0.05 mmol) was dissolved in 3 mL of DCM, 0.3 mL of TFA was added at room temperature and stirred for 2 hours. LCMS showed that the reaction was complete, and the mixture was concentrated under reduced pressure to obtain 57 mg. MS (+) = 1149.6.

[0637] Synthesis of LD-71

[0638] Compound 71-10 (57 mg, 0.050 mmol) was dissolved in 2 mL of anhydrous DMF, and HATU (20.76 mg, 0.055 mmol) was added. The mixture was stirred at room temperature for five minutes. Sulfonic acid quaternary ammonium salt A (15.64 mg, 0.074 mmol, CAS No. 78276-19-4, provided by Shanghai Haoyuan Pharmaceutical Co., Ltd., synthesis method reference WO2011146595A2) and DIEA (0.025 mL, 0.149 mmol) were added sequentially. Stirring was continued at room temperature for 2 hours. LCMS indicated the reaction was complete. Reverse phase preparation (mobile phase containing 0.1% formic acid) was used to obtain 24.0 mg of a white solid.

[0639] LCMS: m / z(ES+)(M+H)+=1341.6, Rt=1.210min

[0640] 1H NMR(400MHz,DMSO)δ10.04-9.96(m,1H),8.45(s,1H),8.185-8.085(m,2H),7.968-7.7 82(m,3H),7.63-7.32(m,9H),6.99(s,2H),6.53(s,1H),5.99(s,1H),5.70(d,J=20.6Hz ,1H),5.43-5.41(m,5H),5.35(s,3H),4.35-4.25(m,1H),4.19-4.17(m,1H),3.07-2.9 1(m,20H),1.97-1.83(m,8H),1.52-1.42(m,6H),1.32-1.15(m,9H),0.90-0.82(m,9H).

[0641] Example 10 (Preparation of LD-72)

[0642] The synthetic route of LD-72 is as follows:

[0643] Synthesis of 72-2

[0644] Compound 72-1 (600 mg, 1.076 mmol) was dissolved in 4 mL of anhydrous DMF, and 0.1 mL of piperidine was added. The mixture was stirred at room temperature for one hour, and then evaporated under reduced pressure and slurried with petroleum ether to obtain 350 mg of crude product. MS+Na(+)=357.9.

[0645] Synthesis of 72-3

[0646] Compound 72-2 (360 mg, 1.073 mmol) and 6-maleimidocaproic acid (340 mg, 1.610 mmol) were dissolved in 10 mL of anhydrous DMF. DIEA (0.532 mL, 3.219 mmol) and HATU (817 mg, 2.150 mmol) were added and stirred at room temperature for 2 hours. LCMS indicated the reaction was complete. Water was added and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed using a normal phase silica gel column (MeOH / DCM, 0% to 5%) to yield 400 mg. MS(+) = 529.6.

[0647] Synthesis of 72-4

[0648] Compound 72-3 (90 mg, 0.071 mmol) was dissolved in 3 mL of anhydrous DCM. TFA (0.8 mL) was added to the reaction mixture and stirred at room temperature for 18 hours. The mixture was concentrated to obtain 80 mg of crude product, which was used directly in the next reaction. MS(+) = 473.5.

[0649] Synthesis of 72-5

[0650] Compound 72-4 (150 mg, 0.317 mmol) was dissolved in 2 mL, HOSu (40.2 mg, 0.349 mmol) and EDCI (67.2 mg, 0.351 mmol) were added, and the mixture was stirred at room temperature for 2 hours. The mixture was then distilled under reduced pressure to obtain 160 mg of crude product, which was used directly in the next reaction. MS(+) = 569.8.

[0651] Synthesis of 72-6

[0652] Compound 65-7 (700 mg, 2.628 mmol) and Fmoc-glycine (1.56 g, 5.25 mmol) were dissolved in 20 mL of anhydrous THF. EEDQ (780 mg, 3.158 mmol) was added and stirred at room temperature for 18 hours. LCMS indicated the reaction was complete. Water was added and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed using a normal phase silica gel column (MeOH / DCM, 0% to 5%) to yield 1000 mg. MS+Na(+) = 567.8.

[0653] Synthesis of 72-7

[0654] Compound 72-6 (290 mg, 0.531 mmol) was dissolved in 6 mL of anhydrous THF, and p-nitrophenyl chloroformate (129 mg, 0.640 mmol) and pyridine (63 mg, 0.797 mmol) were added sequentially. After reacting at room temperature for 3 hours, the reaction solution was added with water and extracted three times with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed using a normal phase silica gel column (MeOH / DCM, 0% to 5%) to yield 300 mg. MS+Na(+) = 732.7.

[0655] Synthesis of 72-8

[0656] Compound 72-7 (340 mg, 0.478 mmol) and isotecan mesylate (254 mg, 0.478 mmol) were dissolved in 8 mL of anhydrous DMF. DMAP (8 mg, 0.025 mmol) and DIEA (0.237 mL, 1.434 mmol) were added sequentially. After reacting at room temperature for 12 hours, the reaction mixture was added with water and extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed using a normal phase silica gel column (MeOH / DCM, 0% to 10%) to yield 380 mg. MS(+) = 1006.7.

[0657] Synthesis of 72-9

[0658] Compound 72-8 (300 mg, 0.298 mmol) was dissolved in 3 mL of anhydrous DMF, and 0.06 mL of piperidine was added. The mixture was stirred at room temperature for one hour, and then evaporated under reduced pressure and slurried with petroleum ether to obtain 230 mg of crude product. MS(+) = 785.8.

[0659] Synthesis of 72-10

[0660] Compound 72-9 (180 mg, 0.316 mmol) and compound 72-5 (207 mg, 0.264 mmol) were dissolved in 3 mL of anhydrous DMF, DIEA was added, and the mixture was stirred at room temperature for 2 hours. Water was added to the reaction mixture, and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed on a thin-layer silica gel plate (MeOH / DCM, 0% to 10%) to yield 100 mg. MS(+) = 1240.8.

[0661] Synthesis of 72-11

[0662] Compound 71-10 (90 mg, 0.071 mmol) was dissolved in 3 mL of anhydrous DCM. TFA (0.3 mL) was added to the reaction mixture and stirred at room temperature for 1 hour. The mixture was concentrated to obtain 36 mg of crude product, which was used directly in the next reaction. MS(+) = 1138.6.

[0663] Synthesis of LD-72

[0664] Compound 71-11 (36 mg, crude) and quaternary ammonium phosphate (11.4 mg, 0.047 mmol, Cas No.: 853798-56-8, provided by Shanghai Haoyuan Pharmaceutical Co., Ltd., synthesis method reference WO2005114193) were dissolved in 3 mL of anhydrous DMF. DIEA (0.015 mL, 0.093 mmol) and HATU (24 mg, 0.032 mmol) were added. The mixture was stirred at room temperature for 3 hours. LCMS indicated the reaction was complete. Purification was performed by reverse phase preparative chromatography (mobile phase containing 10 mM NH4OAc) to yield 15 mg of a white solid.

[0665] LCMS: m / z(ES+)(M+H)+=1361.5, Rt=1.114min

[0666] 1HNMR(400MHz,DMSO)δ10.01(d,J=12Hz,1H),9.04(d,J=8.0Hz,1H),8.40(s,2H),8.27-8.07(m,3H),7 .79-7.60(m,2H),7.48-7.11(m,9H),6.99(s,2H),6.52(s,1H),5.57-5.00(m,8H),4.94-4.37(m,5H),4 .25-4.09(m,2H),3.99-3.48(m,10H),3.08(d,J=13.6Hz,7H),2.94(d,J=13.2Hz,9H),2.40-2.33(m,3 H), 2.19-2.07 (m, 4H), 1.91-1.81 (m, 2H), 1.52-1.41 (m, 4H), 1.33-1.00 (m, 3H), 0.88 (t, J = 7.2Hz, 3H).

[0667] Example 11 (Preparation of LD-73)

[0668] The synthetic route of LD-73 is as follows:

[0669] Synthesis of LD-73

[0670] Compound 73-1 (25.19 mg, 0.055 mmol) and HATU (20.98 mg, 0.055 mmol) were dissolved in 2 mL of anhydrous DMF. After stirring at room temperature for 5 minutes, compound 65-13 (60 mg, 0.050 mmol) and DIEA (0.008 mL, 0.075 mmol) were added sequentially. After two hours, the reaction was complete as monitored by LCMS. The reaction system was then directly subjected to reverse phase preparative chromatography (mobile phase containing 0.1% formic acid) to yield 19 mg of a white solid.

[0671] LCMS: m / z(ES+)(M+H)+=1634.8, Rt=1.392min

[0672] 1H NMR (400MHz, DMSO) δ9.96 (d, J = 40.0Hz, 1H), 8.17 (d, J = 6.8Hz, 1H), 8.00 (s, 1H), 7.78 (d, J = 10. 8Hz,2H),7.62(s,1H),7.38-7.26(m,4H),6.99(s,2H),6.52(s,1H),5.44(s,3H),5.29(s,5H),5 .13-5.10(m,3H),4.67(s,1H),4.58(s,1H),4.36(d,J=7.2Hz,1H),4.22-4.19(m,1H),3.66-3.5 5(m,10H),3.52-3.40(m,50H),3.23(s,6H),3.18-3.10(m,5H),1.24(s,3H),0.89-0.81(m,9H).

[0673] Example 12 (Preparation of LD-74)

[0674] The synthetic route of LD-74 is as follows:

[0675] Synthesis of 74-1

[0676] 6-Maleimidocaproic acid (18.59 mg, 0.088 mmol) was dissolved in 1 mL of anhydrous DMF, and HATU (33.46 mg, 0.088 mmol) was added. The mixture was stirred at room temperature for five minutes. Compound 65-11 (72.11 mg, 0.080 mmol) and DIEA (0.020 mL) were added sequentially. Stirring was continued at room temperature for two hours. LCMS indicated the reaction was complete. Water was added, and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified using a normal phase silica gel column (MeOH / DCM, 0% to 10%) to yield 80 mg. MS(+) was 1091.6.

[0677] Synthesis of 74-2

[0678] Compound 74-1 (65 mg, 0.073 mmol) was dissolved in 3 mL of DCM, 0.6 mL of TFA was added at room temperature and stirred for 2 hours. LCMS showed that the reaction was complete, and the mixture was concentrated under reduced pressure to obtain 65 mg. MS (+) = 991.4.

[0679] Synthesis of LD-74

[0680] Compound 74-3 (20.00 mg, 0.026 mmol, purchased from Shanghai Haoyuan Pharmaceutical Co., Ltd.; synthesis method reference WO2022228493) was dissolved in 1 mL of anhydrous DMF. HATU (10.85 mg, 0.029 mmol) was added, and the mixture was stirred at room temperature for five minutes. Compound 74-2 (25.71 mg, 0.026 mmol) and DIEA (0.008 mL, 0.046 mmol) were added sequentially, and stirring was continued at room temperature for 2 hours. LCMS indicated the reaction was complete. The reaction mixture was prepared using reverse phase chromatography (mobile phase containing 10 mM NH4OAc) to yield 9.5 mg of a white solid.

[0681] LCMS: m / z(ES+)(M+H)+=1743.9, Rt=1.325min

[0682] 1H NMR (400MHz, DMSO) δ7.79(d,J=10.9Hz,2H),7.31(s,1H),6.99(s,2H),6.52(s,1H),5.45(s,2H),5.28(s,3H),5.12(s ,2H),4.33-4.04(m,26H),2.98-2.66(m,45H),1.90(m,7H),1.49-1.47(m,5H),1.31-1.15(m,7H),0.89-0.81(m,9H).

[0683] Example 13 (Preparation of LD-88)

[0684] The synthetic route of LD-88 is as follows:

[0685] Synthesis of LD-88

[0686] Compound 38-8 (50 mg, 0.041 mmol, for its synthesis, see Example 3) and quaternary ammonium sulfonate C (16.34 mg, 0.078 mmol, CN115521998, Shanghai Haoyuan Pharmaceutical Co., Ltd.) were dissolved in 2 mL of anhydrous DMF. DIEA (0.010 mL, 0.062 mmol) and HATU (17.27 mg, 0.045 mmol) were added. The mixture was stirred at room temperature for 2 hours. LCMS indicated the reaction was complete. The reaction mixture was directly purified by reverse phase preparative (trifluoroacetic acid) to obtain LD-88 as a pale yellow solid (10.0 mg, 17.28% yield).

[0687] LCMS: m / z(ES+)(M+H)+=1402.6, Rt=1.303min

[0688] 1H NMR (400MHz, DMSO) δ10.06(s,1H),9.95(s,1H),8.19(d,J=7.6Hz,1H),8.10-8.00(m,2H),7.91(d,J=8Hz,1 H),7.79(d,J=10.8Hz,1H),7.61(d,J=8.4Hz,2H),7.38(d,J=8.4Hz,2H),7.32(s,1H),7.00(s,2H),6.53(s, 1H),5.45(s,2H),5.29(s,3H),5.09(s,2H),4.45-4.41(m,2H),4.20-3.99(m,2H),3.62-3.57(m,4H),3.51- 3.47(m,14H),3.19-3.13(m,5H),2.67(dt,J=9.2,4.4Hz,4H),2.39(s,5H),2.36-2.31(m,5H),2.19(s,3H), 2.06-1.82(m,10H),0.90-0.84(m,9H).

[0689] Example 14 (Preparation of LD-89)

[0690] The synthetic route of LD-89 is as follows:

[0691] Synthesis of LD-89

[0692] Compound 38-8 (50 mg, 0.041 mmol, for its synthesis method, see Example 3) and quaternary ammonium sulfonate D (17.58 mg, 0.078 mmol, Bioconjugate Chem. 2022, 33, 718-725, Shanghai Haoyuan Pharmaceutical Co., Ltd.) were dissolved in 2 mL of anhydrous DMF. DIEA (0.010 mL, 0.062 mmol) and HATU (17.27 mg, 0.045 mmol) were added. The mixture was stirred at room temperature for 2 hours. LCMS showed the reaction was complete. The reaction solution was directly purified by reverse phase preparative (formic acid) to obtain LD-89 as a white solid (5.4 mg, 10.06% yield).

[0693] LCMS: m / z(ES+)(M+H)+=1418.6, Rt=1.288min

[0694] 1HNMR (400MHz, DMSO) δ9.86 (s, 1H), 8.11 (d, J = 7.2Hz, 1H), 8.01-7.89 (m, 3H), 7.71 (d, J = 10.8Hz, 1H), 7.55 (d ,J=8.4Hz,2H),7.30(d,J=8.4Hz,2H),7.24(s,1H),6.93(s,2H),6.45(s,1H),5.38(s,2H),5.22(d,J=4.0Hz,3 H),5.01(s,2H),4.35-4.30(m,1H),4.15-4.12(m,1H),3.52(s,8H),3.42(d,J=3.6Hz,17H),3.25(s,5H),3.0 9-2.99(m,14H),2.31(s,4H),2.26(d,J=7.2Hz,3H),2.16-2.07(m,2H),1.88-1.82(m,8H),0.82-0.76(m,9H).

[0695] Example 15 (Preparation of LD-94)

[0696] The synthetic route of LD-94 is as follows:

[0697] Synthesis of compound 94-1

[0698] Compound 38-5 (450 mg, 0.412 mmol, the synthesis method can be described in Example 3) was added to dichloromethane (5 mL) and trifluoroacetic acid (1 mL) was added dropwise to the system. The mixed system was stirred at room temperature for 2 hours. The system was directly concentrated to dryness and the crude product was used in the next step. m / z (ES+) (M+1) = 1035.4

[0699] Synthesis of compound 94-2

[0700] Compound 94-1 (400 mg, 0.386 mmol), quaternary ammonium sulfonate A (121.90 mg, 0.580 mmol) and DIEA (74.91 mg, 0.580 mmol) were dissolved in DMF (4 mL). The mixture was stirred at room temperature for 5 minutes, and HATU (161.63 mg, 0.425 mmol) was added. The reaction system was stirred at room temperature for 4 hours and then purified by reverse phase column (water:acetonitrile from 0% to 70%) to obtain 170 mg, m / z (ES+) (M+1) = 1227.6.

[0701] Synthesis of compound 94-3

[0702] Compound 94-2 (70 mg, 0.057 mmol) was dissolved in DMF (1 mL), and diethylamine (14.30 mg, 0.196 mmol) was added dropwise. The mixture was stirred at room temperature for 2 hours, concentrated to dryness, slurried with petroleum ether, and filtered. The filter cake was dried and used directly in the next step. m / z (ES+) (M+1) = 1005.6.

[0703] Synthesis of LD-94

[0704] Compound 94 (55 mg, 0.055 mmol), BCN-PEG4-acid (cas number: 1881221-47-1, 24.16 mg, 0.055 mmol) and DIEA (0.011 mL, 0.067 mmol) were dissolved in DMF (2 mL) and stirred at room temperature for 5 minutes. HATU (18.73 mg, 0.049 mmol) was added and the mixture was reacted at room temperature for 2 hours. The product was analyzed by Prep-HPLC (NH4HCO3 system) to give LD-94 (6 mg, yield: 7.68%).

[0705] LCMS: m / z (ES+) (M+H) + =1427.6,Rt=1.721min.

[0706] 1H NMR (400MHz, DMSO) δ9.90(s,1H),8.19(s,2H),8.09-8.05(m,1H),8.00-7.96(m,1H),7.79(d,J=11.2Hz,1H),7.63(d,J=7.6 Hz,2H),7.37(d,J=8.2Hz,2H),7.31(s,1H),6.53(s,1H),5.45(s,1H),5.29(s,3H),5.08(s,2H),4.19-4.15(m,1H),4.02(d, J=8.6Hz,2H),3.62-3.58(m,2H),3.50-3.47(m,16H),3.10(s,2H),3.04-3.02(m,7H),2.69-2.66(m,4H),2.40-2.36(m,5H) ,2.34-2.32(m,4H),2.22-2.15(m,9H),1.99(s,4H),1.92-1.86(m,3H),1.55-1.46(m,2H),1.23(s,2H),0.90-0.83(m,14H).

[0707] Example 16 (Preparation of LD-98)

[0708] The synthetic route of LD-98 is as follows:

[0709] Synthesis of compound 98-1

[0710] Compound 65-11 (120 mg, 0.134 mmol, for its synthesis method, see Example 5) and Fmoc-NMe-PEG4-C2-Acid (Cas No.: 2170240-98-7, 67.0 mg, 0.134 mmol) were dissolved in DMF (2 mL). HATU (55.9 mg, 0.147 mmol, 1.1 eq) and N,N-diisopropylethylamine (26 mg, 0.200 mmol, 1.5 eq) were added sequentially, and the mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM:MeOH, MeOH 0 to 10%) to obtain 100 mg of the product, m / z (ES+)(M+1) = 1381.6.

[0711] Synthesis of compound 98-2

[0712] Compound 98-1 (100 mg, 0.072 mmol) was dissolved in DMF (2 mL), and diethylamine (15.88 mg, 0.217 mmol) was added, followed by stirring at room temperature for 2 hours. The mixture was concentrated under reduced pressure to obtain 100 mg of a crude product, m / z (ES+) (M+1) = 1159.6.

[0713] Synthesis of compound 98-3

[0714] Compound 98-2 (80 mg, 0.069 mmol) and 4-methylsulfonylbenzoic acid (13.95 mg, 0.069 mmol) were dissolved in DMF (2 mL). HATU (28.86 mg, 0.076 mmol) and DIPEA (13.38 mg, 0.104 mmol) were added sequentially, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the residue was purified by reverse phase chromatography (C18, 20-60% acetonitrile-H2O solution) to give 26 mg of a yellow solid, m / z (ES+) (M+1) = 1343.49.

[0715] Synthesis of compound 98-4

[0716] Compound 98-3 (25 mg, 0.019 mmol) was dissolved in DCM (2 mL) and TFA (0.4 mL) was added. The reaction was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure to obtain crude compound 18 (23 mg), which was used directly in the next reaction. m / z (ES+) (M+1) = 1243.4.

[0717] Synthesis of LD-98

[0718] Compound 98-4 (23 mg, 0.018 mmol) and quaternary ammonium sulfonate B (7.03 mg, 0.028 mmol) were dissolved in DMF (1 mL). HATU (7.74 mg, 0.020 mmol) and DIPEA (3.59 mg, 0.028 mmol) were added sequentially. The reaction mixture was stirred at room temperature for 2 hours. The reaction system was purified by pre-HPLC (TFA) to afford LD-98 (6 mg, 22.2% yield).

[0719] LCMS: m / z (ES+) (M+H) + =1479.6,Rt=1.340min.

[0720] 1H NMR(400MHz, DMSO)δ9.97(d,J=40.2Hz,1H),9.14(d,J=12.0Hz,2H),8.19(t ,J=7.4Hz,1H),8.06(d,J=9.4Hz,1H),7.85(s,1H),7.78(d,J=11.2Hz,1H),7 .64(dd,J=49.2,8.4Hz,1H),7.33(d,J=12.4Hz,2H),6.52(s,1H),5.44(d,J= 4.0Hz,2H),5.30(s,3H),5.12(d,J=10.8Hz,2H),4.64(d,J=26.4Hz,2H),4.3 6(dd,J=13.6,6.8Hz,1H),4.23-4.14(m,1H),3.65(s,1H),3.59-3.54(m,3H) ,3.51-3.46(m,15H),3.43(s,4H),3.03(d,J=9.2Hz,7H),2.98(s,3H),2.92( s,2H),2.86(s,1H),2.68-2.67(m,1H),2.38(s,5H),2.33(dd,J=3.6,1.8Hz, 2H),2.20(s,2H),1.98-1.83(m,8H),1.31-1.23(m,4H),0.89-0.81(m,10H).

[0721] Example 17 (Preparation of LD-99)

[0722] The synthetic route of LD-99 is as follows:

[0723] Synthesis of Compound 99-1

[0724] Exo-3,6-epoxy-1,2,3,6-tetrahydrophthalic anhydride (CAS No. 6118-51-0, 2 g, 12.04 mmol, 1.0 eq) and N-Boc-N-methylethylenediamine (CAS No. 121492-06-6, 2.52 g, 14.46 mmol, 1.2 eq) were dissolved in methanol (60 mL). Triethylamine (2.43 g, 24.1 mmol, 2.0 eq) was added at room temperature. The reaction solution was heated to 65°C and reacted for 48 h. The reaction solution was concentrated and diluted with ethyl acetate (60 mL), dried over saturated brine and anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (petroleum ether:ethyl acetate = 1:1) to obtain 3 g of a white solid. m / z (ES+)(M+H) + =323.3.

[0725] Synthesis of compound 99-2

[0726] Compound 99-1 (2.3 g, 7.135 mmol) was dissolved in dichloromethane (20 mL), trifluoroacetic acid (4 mL) was added in an ice-water bath, and the reaction solution was warmed to room temperature and reacted for 1 hour. The reaction solution was concentrated to obtain 1.5 g of crude product, m / z (ES+) (M+H) + =223.2.

[0727] Synthesis of compound 99-3

[0728] Compound 99-2 (1.58 g, 7.11 mmol, 1.0 eq) and tert-butyl 6-bromohexanoate (2.14 g, 8.52 mmol, 1.2 eq) were dissolved in DMF (10 mL) and reacted at room temperature for 72 hours. The reaction solution was concentrated and then purified by column chromatography (petroleum ether:ethyl acetate = 3:1) to obtain 2.5 g of the desired product, m / z (ES+)(M+Na) + =393.2.

[0729] Synthesis of compound 99-4

[0730] Compound 99-3 (1.5 g, 3.822 mmol) was dissolved in toluene (15 mL) and heated to 110°C for 18 hours. The reaction solution was concentrated to obtain the desired product as a yellow oil (1 g). m / z (ES+) (M+H) + =325.2.

[0731] Synthesis of compound 99-5

[0732] Compound 99-4 (100 mg, 0.308 mmol) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (0.7 mL) was added, and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated to obtain 82 mg of crude product as a yellow oil, m / z (ES+) (M+H) + =269.2.

[0733] Synthesis of Compound 99-6

[0734] Compound 65-11 (88 mg, 0.098 mmol, 1.0 eq) and compound 99-5 (78.8 mg, 0.294 mmol, 3.0 eq) were dissolved in DMF (3 mL), and HATU (112 mg, 0.294 mmol, 3.0 eq) and diisopropylethylamine (0.081 mL, 0.489 mmol, 5.0 eq) were added in sequence. The mixture was reacted at room temperature for 2 hours. The reaction solution was filtered and purified by reverse column chromatography to obtain 80 mg of a yellow solid, m / z (ES+) (M+H) + =1147.7.

[0735] Synthesis of compound 99-7

[0736] Compound 99-6 (80 mg, 0.070 mmol) was dissolved in dichloromethane (3 mL), trifluoroacetic acid (0.3 mL) was added dropwise, and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated to obtain 73 mg of crude product as a yellow oil, m / z (ES+) (M+H) + =1047.6.

[0737] Synthesis of LD-99

[0738] Compound 99-7 (73 mg, 0.070 mmol, 1.0 eq) and quaternary ammonium sulfonate B (35 mg, 0.14 mmol, 2.0 eq) were dissolved in DMF (2 mL). HATU (53 mg, 0.14 mmol, 2.0 eq) and N,N-diisopropylethylamine (0.058 mL, 0.35 mmol, 5.0 eq) were added sequentially. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was filtered and purified to obtain 9.2 mg of a white solid.

[0739] LCMS: m / z (ES+) (M+H) + =1284.6,Rt=1.217min

[0740] 1 H NMR (400MHz, DMSO) δ10.01(d,J=38.5Hz,1H),9.24(s,1H),8.26-8.16(m,1H),8.13-8.01(m,1H),7.80(t,J=9.2Hz,2H),7.64(dd ,J=47.3,8.1Hz,1H),7.41-7.31(m,3H),7.17(s,1H),7.10(d,J=4.1Hz,2H),6.54(s,1H),5.44(d,J=4.0Hz,2H),5.29(s,3H),5. 13(d,J=8.2Hz,2H),4.64(d,J=27.5Hz,2H),4.36(d,J=7.5Hz,1H),4.22-4.16(m,1H),3.77(s,2H),3.22(s,2H),3.05(s,5H),2. 99(d,J=3.8Hz,4H),2.94(s,2H),2.87(s,2H),2.81(s,4H),2.36(d,J=18.7Hz,5H),2.19(dd,J=13.7,7.2Hz,5H),2.06-1.84(m, 9H),1.53(s,5H),1.34-1.21(m,7H),0.89-0.83(m,9H).

[0741] Test Example 18 (Preparation of the compound of formula (2))

[0742] Coupling method 1 (full reduction, drug-antibody ratio of 8)

[0743] The antibody solution was exchanged into a pH 7.4 phosphate buffer using an ultrafiltration centrifuge tube, and TCEP at 10-15 times the molar number of the antibody was added. The solution was reacted at 37°C for 2 hours to open the interchain disulfide bonds of the antibody. After the reduction was completed, the antibody solution was exchanged into a pH 7.4 phosphate buffer using an ultrafiltration centrifuge tube to remove excess TCEP. Linker-Drug (the compound of formula (1), compound MC-GGFG-Exatecan, compound MC-GGFG-Dxd, and compound ELC122 prepared in Examples 1-5, 7-17) at 12-20 times the molar number of the antibody was added and reacted at room temperature for 1 hour. After the reaction was completed, cysteine ​​at 2 times the molar number of the linker-drug was added to quench the reaction, and the solution was exchanged into a pH 5.5 acetate buffer using an ultrafiltration centrifuge tube. The antibody-drug conjugate sample after the solution exchange was filtered using a 0.22 micron sterilizing filter and then used.

[0744] Coupling method 2 (partial reduction, drug-antibody ratio of 4)

[0745] Use an ultrafiltration centrifuge tube to exchange the antibody solution into a pH 7.4 phosphate buffer, add 2.2-2.3 times the number of molar molecules of the antibody TCEP, and react at 37 ° C for 2 hours to open the disulfide bonds between the antibody chains. After the reduction is completed, add 6-8 times the number of molar molecules of the antibody Linker-Drug (the compound of formula (1) and the compound MC-VC-PAB-MMAE prepared in Example 6) and react at room temperature for 1 hour. After the reaction is completed, add 2 times the number of molar molecules of the Linker-Drug cysteine ​​to quench the reaction, and use an ultrafiltration centrifuge tube to exchange the solution into a pH 5.5 acetate buffer. The antibody drug conjugate sample after the solution exchange is filtered using a 0.22 micron sterilizing filter for use.

[0746] Conjugated antibody sequence

[0747] The amino acid sequence of the heavy chain of the anti-claudin 18.2 humanized monoclonal antibody 9D1 is as follows:

[0748] The amino acid sequence of the light chain of the anti-claudin 18.2 humanized monoclonal antibody 9D1 is as follows:

[0749] The heavy chain amino acid sequence of the anti-human folate receptor (FRa) humanized monoclonal antibody Farletuzumab is as follows:

[0750] The amino acid sequence of the light chain of the anti-human folate receptor (FRa) humanized monoclonal antibody Farletuzumab is as follows:

[0751] Chromatographic analysis of antibody drug conjugates

[0752] Size exclusion chromatography (SEC) analysis was performed on a Tskgel G3000SWXL column (TOSOH, 0008541) with a mobile phase of 0.1 M PB, pH 7.0, 0.1 M NaCl. Isocratic elution was performed at a flow rate of 1 mL / min and a column temperature of 25°C. Detection wavelengths were 280 nm and 366 nm. Hydrophobic interaction chromatography (HIC) analysis was performed on a Butyl-NPR 4.6 × 100 mm column (TOSOH, 0042168) with mobile phase A of 50 mM PB, pH 7.0, 1.5 M (NH₄)₂SO₄ and mobile phase B of 50 mM PB, pH 7.0, 20% acetonitrile. The separation gradient was 0% B to 100% B over 2–20 minutes. The column temperature was 30°C and the flow rate was 0.6 mL / min. The detection wavelengths were 280 nm and 366 nm, and the results are shown in Table 1 and Figures 1-6 (Figure 1 includes Figure 1A and Figure 1B, Figure 2 includes Figure 2A and Figure 2B, Figure 3 includes Figure 3A, Figure 3B, Figure 3C, Figure 3D, Figure 3E, Figure 3F, Figure 3G, and Figure 3H, Figure 4 includes Figure 4A and Figure 4B, Figure 5 includes Figure 5A and Figure 5B, Figure 6 includes Figure 6A, Figure 6B, Figure 6C, Figure 6D, Figure 6E, Figure 6F, Figure 6G, and Figure 6H).

[0753] Table 1 shows the molecular sieve exclusion chromatography (SEC) and hydrophobic interaction chromatography (HIC) of each test example and comparative example.

[0754] Summary: As shown in Table 1 and Figures 1-6, the anti-claudin 18.2 humanized monoclonal antibody 9D1 (WO2021160155) was used as a model antibody to couple with the designed linker drug compound and the control linker drug compound to prepare antibody-drug conjugates. The linker drug compounds MC-GGFG-Exatecan (Shanghai Haoyuan Biotechnology, Catalog No. HY-114233), MC-VC-PAB-MMAE (Shanghai Haoyuan Biotechnology, Catalog No. 646502-53-6), and MC-GGFG-Dxd (Deruxtecan) (Shanghai Haoyuan Biotechnology, Catalog No. HY-13631E) were purchased; LD-74 was synthesized using the synthesis method of WO2022228493; and ELC122 was synthesized using the synthesis method of WO2021225892.

[0755] Size exclusion chromatography (SEC) analysis of the resulting 9D1 antibody-drug conjugates demonstrated SEC purity exceeding 97%, regardless of whether the conjugated drug was exatecan, SN38, or MMAE. The purity of the control linker LD-27 antibody-drug conjugate was only 89.22% (Table 1).

[0756] The results of hydrophobic interaction chromatography (HIC) analysis of 9D1 antibody-drug conjugates showed that the internal salt linkers designed for toxins MMAE, SN38 and Exatecan significantly shortened the HIC retention time of 9D1 antibody-drug conjugates compared with the control linkers (excluding LD-74). The retention time of the main peak (antibody drug conjugate ratio of 4) of the inner salt linker-MMAE antibody drug conjugate (herein referring to the antibody drug conjugate of LD-57) is shortened by about 1 minute (16.07 vs 16.99) compared with the control linker-MMAE (MC-VC-PAB-MMAE) antibody drug conjugate (herein referring to the antibody drug conjugate of Comparative Example 2), and the retention time is shortened more significantly for components with increased antibody drug conjugate ratios (Table 1, Figure 4); the retention time of the inner salt linker-SN38 antibody drug conjugate (LD-71 antibody drug conjugate) is shortened by about 4 minutes (12.93 vs 16.74 Table 1, Figure 5) compared with the control linker-SN38 (ELC122, WO2021225892) antibody drug conjugate (herein referring to the antibody drug conjugate of Comparative Example 3). The HIC retention time of the internal salt linker-Exatecan antibody drug conjugates of the present application (LD-31, LD-37, LD-38, LD-65, LD-66, LD-72, LD-88, LD-89, LD-98 and LD-99 antibody drug conjugates) is between 13.12 and 14.17 min, which is significantly shorter than the retention time of the control linker-Exatecan (LD-12, LD-27, MC-GGFG-Exatecan) and MC-GGFG-Dxd antibody drug conjugates without hydrophilic side chains, and is also significantly shorter than the retention time of the LD-73 antibody drug conjugate containing a hydrophilic side chain of polyethylene glycol (PEG 9) (16.00 min), and is comparable to the retention time of the LD-74 antibody drug conjugate containing a hydrophilic side chain of polysarcosine (13.58 min) (Table 1, Figure 6). An interesting finding is that for the inner salt linker, Exatecan, the closer the inner salt is to the exatecan toxin on the linker, the shorter the HIC retention time of the corresponding antibody-drug conjugate. For example, for the sulfonic acid inner salt linker, Exatecan, the corresponding antibody-drug conjugate HIC retention time of LD-37 (sulfonic acid inner salt at position Z of formula (1-1), farther from the exatecan) is 14.07 minutes. The corresponding antibody-drug conjugates of LD-38 (sulfonic acid inner salt at position Z of formula (1-1-1)), LD-65 (sulfonic acid inner salt at position Z'), and LD-31(P2) (sulfonic acid inner salt at position Z") have retention times of 13.57, 13.46, and 13.40 minutes, respectively, which are significantly shorter than the 14.07 minutes of the antibody-drug conjugate corresponding to LD-37 (Table 1, Figure 6).

[0757] Hydrophobic interaction chromatography reveals the hydrophobic characteristics of antibody-drug conjugates. This hydrophobicity is primarily due to the hydrophobicity of the conjugated drug compound. Strong hydrophobicity of the antibody-drug conjugate results in prolonged retention time on HIC. However, the antibody-drug conjugate has poor water solubility and stability, and is easily cleared from the body. The exceptionally strong water solubility of the internal salt linker can significantly mask the hydrophobicity of the drug compound, enhancing the water solubility and stability of the antibody-drug conjugate and shortening its retention time on HIC.

[0758] In addition, conjugates of the internal salt linker-exatecan (LD-38) with the anti-human folate receptor (FRa) humanized monoclonal antibody farletuzumab and the IgG1 isotype control monoclonal antibody (Shanghai Bio-Pharma, Cat. No. B117901) were prepared with SEC purity >96% and HIC retention time of approximately 13 minutes (Table 1), which is comparable to the retention time of the corresponding 9D1 antibody-drug conjugate.

[0759] The 9D1 antibody-drug conjugates (i.e., the compounds represented by formula (2)) in the following experimental examples were all prepared according to Experimental Example 18.

[0760] Test Example 19 (Drug-Antibody Ratio (DAR))

[0761] Sample processing:

[0762] 9D1-LD-38 prepared in Experimental Example 5a was diluted to 1 mg / mL with 0.1 M Tris-HCl. 5 μL of 0.5 M DTT was added per 100 μg of 9D1-LD-38. The mixture was incubated in a 37°C water bath for 30 min, and then 1% (v / v) 10% TFA was added to terminate the reaction.

[0763] LC-MS parameters:

[0764] Mobile phase A consisted of 0.10% TFA and 0.02% FA in water, and mobile phase B consisted of 0.08% FA and 0.02% TFA in acetonitrile. The separation gradient was from 20% to 32% mobile phase B over 2-4 minutes, and from 32% to 45% over 4-12 minutes. The flow rate was 0.4 mL / min. The column was a BioResolve™ RP mAb Polyphenyl Column (2.7 μm, 2.1 × 150 mm), and the column temperature was 70°C.

[0765] The mass spectrometer was a Themo Q Exactive Plus, using the positive ion mode Full Scan, spray voltage 3.8 kV, and scan range 500-3500 m / z.

[0766] RP-HPLC parameters:

[0767] Mobile phase A was 0.1% TFA in water, and mobile phase B was 0.1% TFA in acetonitrile. The separation gradient was 5-25 min, with mobile phase B increasing from 30% to 50%. The flow rate was 0.6 mL / min. The chromatographic column was a Proteomix RP-1000 (5 μm, 4.6 × 100 mm) and the column temperature was 60°C.

[0768] The results are shown in Table 2 and Figures 7-9.

[0769] Table 2 shows the theoretical molecular weight and detected molecular weight of the main components of 9D1-LD-38 LC / MS

[0770] Summary: As shown in Table 2 and Figures 7-9, since 9D1-LD-38 utilizes cysteine ​​conjugation, it underwent a pretreatment to retain glycosylation in its reduced state. The results showed that both the light chain and heavy chain had a single major component. Mass spectrometry molecular weight identification revealed that the primary conjugated form of the light chain was one LD-38 molecule, while the primary conjugated form of the heavy chain was three LD-38 molecules. Molecular weight deconvolution results, shown in Figures 7 and 8, were consistent with the theoretical molecular weight. Partial ring opening of the LD-38 molecule and the presence of heterogeneous heavy chain conjugations were also observed. The DAR value of 9D1-LD-38 was calculated using the area normalization method of the liquid chromatography chromatogram (Figure 9), which is comparable to the theoretical drug-antibody conjugated ratio of 8.

[0771] Test Example 20 (In vitro tumor cell killing activity)

[0772] 1. Human claudin 18.2-positive MC38-hClaudin18.2 cell line

[0773] The 9D1 antibody-drug conjugates prepared in the aforementioned experimental and comparative examples were used to test in vitro cytotoxicity in MC38 cells stably expressing human claudin 18.2. MC38-hClaudin18.2 cells were seeded at 3,000 cells per well in a 96-well cell culture plate and cultured overnight. The next day, serially diluted 9D1 ADCs were added and cultured for 48 to 72 hours. Fluorescence in each well was measured using AlamarBlue reagent (ThermoFisher, Cat. No. DAL1100), and cell viability was calculated.

[0774] MC38-hClaudin18.2 cells were cultured in RPMI 1640 medium containing 10% FBS and 1 μg / mL puromycin in a 37°C, 5% CO2 incubator for adherent culture. When the cell confluence was greater than 80%, the cells were passaged at a ratio of 1:10 to 1:20 and every 3 days.

[0775] MC38-hClaudin18.2 cells were digested with recombinant enzyme digestion solution (TrypLETM Express Enzyme), and after counting, an appropriate amount of cells were taken and resuspended in RPMI 1640 + 10% FBS to adjust the cell density to 3×10 4 The cells were plated at 100 μL / well in a 96-well cell culture plate and incubated overnight at 37°C in a 5% CO2 incubator. The next day, the test drug was serially diluted 10-fold to a total of 7-8 concentration points; a blank control with a sample concentration of 0 was also set up. The diluted sample was added to the 96-well cell culture plate containing cells at 100 μL / well and incubated at 37°C in a 5% CO2 incubator for 48-72 hours. 20 μL of AlamarBlue detection reagent was added to each well and incubated at 37°C for 4-6 hours. The fluorescence signal intensity was measured on a multifunctional microplate reader with an excitation wavelength of 560 nm and an emission wavelength of 590 nm.

[0776] The cell viability of the blank control was set as 100%, and the cell viability (%) of the experimental wells was calculated according to the following formula:

[0777] Cell viability (%) = (fluorescence value of experimental well / fluorescence value of blank control well) × 100%

[0778] Nonlinear regression using the Sigmoidal dose-response (Variable Slope) method (GraphPad Prism software, GraphPad Software, San Diego, California) was performed using the mean cell viability as the ordinate to generate cell proliferation inhibition curves and calculate the corresponding IC50 (Half Maximum Inhibitory Concentration) values. The results are shown in Table 3 and Figure 10 (Figures 10A, 10B, 10C, 10D, and 10E).

[0779] Table 3 shows the half effective concentration (IC50) of 9D1 antibody drug conjugate and control antibody drug conjugate on MC38-hClaudin18.2 cells in vitro

[0780] Summary: As shown in Table 3 and Figure 10, 9D1 antibody-drug conjugates exhibited specific cytotoxicity against MC38-hClaudin18.2, compared to IgG1 isotype control antibody-drug conjugates (Isotype IgG1-LD-12 and Isotype IgG1-LD-38), with IC50 values ​​ranging from 0.7 to 11.03 nM. 9D1-GGFG-Exatecan exhibited the poorest pharmacodynamic activity, with an IC50 of 11.03 nM. 9D1 antibody-drug conjugates containing internal salt hydrophilic side chains exhibited IC50 values ​​ranging from 0.8 to 4.4 nM. With the exception of 9D1-LD-31-P1, all exhibited significantly greater activity than the exatecan small molecule toxin (IC50 value of 4.10 nM).

[0781] Against MMAE toxin, the IC50 value of the IgG1 isotype control antibody drug conjugate (Isotype IgG1-VC-PAB-MMAE) was 41.3 nM against MC38-hClaudin18.2, and the 9D1 antibody drug conjugates (9D1-vc-PAB-MMAE and 9D1-LD-57) showed specific killing with IC50 values ​​of 0.41 nM and 0.53 nM, respectively.

[0782] 2. Human claudin 18.2-positive NUGC4-hClaudin18.2 cell line

[0783] Human gastric cancer cell line NUGC4 stably expressing human claudin 18.2 (NUGC4-hClaudin18.2) cells were cultured in RPMI 1640 medium containing 10% FBS and supplemented with 1 μg / mL puromycin in a 37°C, 5% CO2 incubator for adherent culture. When the cell confluence was greater than 80%, the cells were passaged at a ratio of 1:3 to 1:10 and passaged every 3 to 4 days.

[0784] NUGC4-hClaudin18.2 cells were digested with recombinant enzyme digestion solution (TrypLETM Express Enzyme). After counting, an appropriate amount of cells were taken and resuspended in RPMI 1640 + 10% FBS to adjust the cell density to 3×10 4Cells were plated at 100 μL / well in a 96-well cell culture plate and incubated overnight at 37°C in a 5% CO2 incubator. The next day, 9D1 ADC and small molecule toxins were serially diluted fivefold starting at 1000 nM (final concentration 500 nM) for a total of 7-8 concentration points. A negative control with a sample concentration of 0 and a blank control consisting of only culture medium without cells were also set up. The diluted samples were added to a 96-well cell culture plate containing cells at 100 μL / well (in duplicate) and incubated at 37°C in a 5% CO2 incubator for 120 hours. 20 μL of CCK-8 detection reagent (Dongren Chemical, Cat. No. CK04) was added to each well and incubated at 37°C for 3-4 hours. The OD value of each well was measured on a multi-function microplate reader at a measurement wavelength of 450 nm and a reference wavelength of 630 nm. OD value per well = OD450 nm - OD630 nm. The cell viability of the negative control was set as 100%, and the cell viability (%) of the experimental wells was calculated according to the following formula.

[0785] Cell viability (%) = (OD value of experimental well - OD value of blank well) / (OD value of negative control well - OD value of blank control well) × 100%

[0786] Sigmoidal dose-response (Variable Slope) method (GraphPad Prism software, GraphPad Software, San Diego, California) was used for nonlinear regression to calculate the corresponding IC50 (Half Maximal Inhibitory Concentration). The results are shown in Table 4.

[0787] Table 4 shows the half effective concentration (IC50) of 9D1 antibody drug conjugate and control antibody drug conjugate on NUGC4-hClaudin18.2 cells in vitro

[0788] Summary: As shown in Table 4, 9D1 antibody-drug conjugates exhibited specific cytotoxicity against NUGC4-hClaudin18.2, with IC50 values ​​ranging from 0.18 to 1.27 nM, compared to an IgG1 isotype control antibody-drug conjugate (Isotype IgG1-LD-38, IC50 141.3 nM). 9D1 antibody-drug conjugates containing internal salt hydrophilic side chains exhibited IC50 values ​​(0.55 to 1.00 nM), superior to those containing polyethylene glycol (PEG, 9D1-LD-73, IC50 1.27 nM) and polysarcosine (PSAR, 9D1-LD-74, IC50 1.16 nM) hydrophilic side chains.

[0789] Against MMAE toxin, the IC50 values ​​of the 9D1-LD-57 antibody drug conjugate containing an internal salt hydrophilic side chain and the control antibody drug conjugate 9D1-vc-PAB-MMAE were comparable, at 0.05 nM and 0.06 nM, respectively.

[0790] 3. Human claudin 18.2-positive HEK293-hClaudin18.2 cell line

[0791] HEK293-hClaudin18.2 cells were cultured in DMEM medium containing 10% FBS and supplemented with 250 μg / mL G418 in a 37°C, 5% CO2 incubator for adherent culture. When the cell confluence was greater than 80%, the cells were passaged at a ratio of 1:20 to 1:40 and every 3 days.

[0792] HEK293-hClaudin18.2 cells were digested with recombinant enzyme digestion solution (TrypLETM Express Enzyme), and after counting, appropriate amount of cells were taken and resuspended in DMEM + 10% FBS to adjust the cell density to 2×10 4 / mL, inoculated into 96-well cell culture plates at 100 μL / well, and cultured overnight in a 37°C, 5% CO2 incubator. The next day, 9D1 ADC and small molecule toxins were diluted 10-fold starting from 1000nM (final concentration of 500nM), with a total of 7 to 8 concentration points; at the same time, a negative control with a sample concentration of 0 and a blank control with only the same volume (200 μL / well) of culture medium added without cells were set. The diluted samples were added to 96-well cell culture plates containing cells at 100 μL / well (double wells) and incubated in a 37°C, 5% CO2 incubator for 72 hours. The cell activity test and data processing were performed using the aforementioned AlamarBlue detection method to calculate the IC50 value. The results are shown in Table 5.

[0793] Table 5 shows the in vitro cell killing half effective concentration (IC50) of 9D1 antibody drug conjugate and control antibody drug conjugate on HEK293-hClaudin18.2 cells Note: NA means not applicable

[0794] Summary: As shown in Table 5, for Exatecan toxin, compared with the IgG1 isotype control antibody drug conjugate (Isotype IgG1-LD-38, IC50688.2 nM), the 9D1 antibody drug conjugate containing an internal salt hydrophilic side chain showed specific killing against HEK293-hClaudin18.2, with IC50 values ​​of 0.09-1.00 nM, and the IC50 of the small molecule Exatecan (0.51 nM).

[0795] Targeting the SN38 toxin, the 9D1 antibody-drug conjugate demonstrated specific cytotoxicity against HEK293-hClaudin18.2 compared to an IgG1 isotype control antibody-drug conjugate (Isotype IgG1-ELC122, which showed no cytotoxicity). The 9D1 antibody-drug conjugate containing an internal salt hydrophilic side chain (9D1-LD-57) showed slightly stronger cytotoxicity than the control (9D1-ELC122), with IC50 values ​​of 0.40 and 0.63 nM, respectively.

[0796] Test Example 21 (In vivo tumor therapeutic effect)

[0797] 1. Human Claudin 18.2-positive MC38-hClaudin18.2 cell line mouse transplant tumor model

[0798] C57BL / 6 mice, female, 5-6 weeks old, were purchased from Shanghai Lingchang. The animals were housed in a laminar flow rack in an SPF animal room, 5 per cage, with 12 hours of alternating light, and free access to water and food. The mouse colorectal cancer cell line MC38-Claudin18.2 that stably expresses human claudin 18.2 was cultured in T75 square flasks in RPMI 1640 medium containing 10% FBS (Fetal Bovine Serum), placed in a 37°C, 5% CO2 incubator, and passaged every three days. MC38 h Claudin18.2 cells in the logarithmic growth phase were collected, centrifuged at 1000 rpm for 5 minutes, and resuspended in PBS. 2×10 6 The cells were mixed with matrix gel at a volume ratio of 1:1 and then inoculated subcutaneously into C57BL / 6 mice. 3 The tumor size was measured twice a week and the tumor volume (TV, mm 3 ), and draw the tumor growth curve.

[0799] TV(mm 3 )=(L×W 2 ) / 2, L represents the tumor length, W represents the tumor width

[0800] Tumor inhibition rate (%) = (TV control group - TV administration group) × 100 / TV control group

[0801] GraphPad Prism software (GraphPad Software, San Diego, California) was used to plot mouse tumor curves in XY mode, with the number of days after the first dose as the horizontal axis and the tumor volume as the vertical axis. Two-way ANOVA was used to analyze the statistical differences between each group and the vehicle control PBS group: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

[0802] The treatment regimen of the 9D1 antibody-drug conjugate conjugated with Exatecan in the MC38-hClaudin18.2 mouse subcutaneous tumor model is shown in Table 6, and the tumor treatment effect and corresponding tumor growth curve are shown in Table 6 and Figure 11 (Figure 11 includes Figures 11A, 11B, and 11C).

[0803] Table 6 shows the grouping, dosing regimen, and tumor inhibition rate after drug treatment of the MC38-hClaudin18.2 mouse subcutaneous tumor model. Note: ip: intraperitoneal injection; NA: not applicable.

[0804] Summary: As shown in Table 6 and Figure 11 , at a single 10 mg / kg dose, the control antibody-drug conjugates (9D1-ELC122, 9D1-GGFG-Exatecan, and 9D1-LD-12) slowed tumor growth ( Figure 11A ), with tumor inhibition rates (TGIs) of 39.82%, 48.34%, and 48.63%, respectively, at the end of the experiment. Antibody-drug conjugates containing internal salt groups in their linkers (9D1-LD-31-P2 and 9D1-LD-38) exhibited superior tumor therapeutic effects ( Figure 11B ), with TGIs of 80.0% and 80.6%, respectively.

[0805] At 10 mg / kg, administered once weekly for two doses, the control antibody drug conjugate 9D1-GGFG-Dxd achieved a tumor inhibition rate of 50.0% at the end of the experiment, while 9D1-LD-38 tumors were almost completely eliminated (tumor inhibition rate 98.0%). The isotype control antibody drug conjugate (IgG1 isotype-LD-38) exhibited only a slight tumor inhibition effect (Table 6).

[0806] To further compare and differentiate the tumor therapeutic efficacy of other 9D1 antibody-drug conjugates containing internal salt groups, a low-dose (5 mg / kg) pharmacodynamic study was conducted, with 9D1-LD-38 serving as an internal reference. At the low-dose (5 mg / kg) dose, the isotype control antibody-drug conjugate (IgG1 isotype-LD-38) showed no significant tumor therapeutic effect. 9D1-LD-38 demonstrated a significant tumor therapeutic effect, with a TGI of 39.2%. Other 9D1 antibody-drug conjugates containing internal salt groups in their linkers (9D1-LD-65, 9D1-LD-66, and 9D1-LD-72) demonstrated even better tumor therapeutic efficacy (Figure 11C), with TGIs of 71.7%, 48.0%, and 42.3%, respectively. In particular, 9D1-LD-65 exhibited the best tumor inhibition effect, with the tumor inhibition growth curve ( Figure 11C ) and TGI significantly superior to those of reported 9D1 antibody-drug conjugates containing other water-soluble groups on the linker, such as the 9D1-LD-73 (TGI 47.0%) control containing a polyethylene glycol (PEG) water-soluble group and the 9D1-LD-74 (TGI 56.5%) control containing a polysarcosine (PSAR) water-soluble group.

[0807] In summary, 9D1 ADCs containing water-soluble groups on the linker (9D1-LD-31-P2, 9D1-LD-38, 9D1-LD-65, 9D1-LD-66, and 9D1-LD-72) showed significantly better efficacy than classic ADCs without linker-enhancing groups, such as 9D1-ELC122, 9D1-GGFG-Exatecan, and 9D1-LD12. In particular, 9D1-LD-65, 9D1-LD-66, and 9D1-LD-72 demonstrated comparable or superior tumor therapeutic efficacy to 9D1 ADCs containing other types of water-soluble groups on the linker (9D1-LD-73 containing PEG and 9D1-LD-74 containing PSAR), such as 9D1-LD-65.

[0808] 2. Human Claudin 18.2-positive NUGC4-hClaudin18.2 cell line mouse transplant tumor model

[0809] The gastric cancer cell line NUGC4-hClaudin18.2 stably expressing human claudin 18.2 was cultured in RPMI-1640 medium containing 10% fetal bovine serum, 100 U / ml penicillin, and 100 μg / mL streptomycin at 37°C in a 5% CO2 incubator. The cells were passaged every 4 to 5 days after confluence. Tumor cells in the logarithmic growth phase were used for inoculation of tumors in vivo.

[0810] NUGC4-hClaudin18.2 tumor cells were adjusted to a concentration of 5×10 7 / mL, inoculated subcutaneously in the right flank of BALB / c nu / nu mice (female, 6-7 weeks old), 100 μL / mouse, and the tumor cell inoculation volume was 5×10 6 / mouse. When the tumor volume grows to about 150mm 3 The groups and dosing schedules are shown in Table 7. Tumor size was measured twice a week and tumor volume (TV, mm 3 ), and draw the tumor growth curve.

[0811] TV(mm 3 )=(L×W 2 ) / 2, L represents the tumor length, W represents the tumor width

[0812] The mouse tumor curve was drawn using GraphPad Prism software (GraphPad Software, San Diego, California) in XY mode, with the number of days after the first administration as the horizontal axis and the tumor volume as the vertical axis. The results are shown in FIG12 .

[0813] Table 7 shows the grouping and dosing regimen of the NUGC4-hClaudin18.2 mouse subcutaneous tumor model. Note: ip: intraperitoneal injection; QW X 3: administration once a week, 3 times.

[0814] Summary: Table 7 and Figure 12 show that at a 5 mg / kg dose, administered once weekly for three times, 9D1-LD-38, whose linker contains an internal salt-promoting water-solubilizing group, exhibited the best therapeutic effect, significantly outperforming both 9D1-ECL122 and 9D1-GGFG-Exatecan. Tumors in the 9D1-LD-38 group were essentially eliminated by day 24 of treatment, while tumor growth was inhibited in the 9D1-GGFG-Exatecan group, but no tumor shrinkage was observed. 9D1-ECL122 exhibited only a weak tumor-suppressing effect, likely due to the fact that its toxin is the less toxic SN38.

[0815] 3. Folate receptor-positive OVCAR-3 cell mouse transplant tumor model

[0816] OVCAR-3 (OVCAR3) human ovarian cancer cells expressing folate receptor (FRa) were cultured in RPMI-1640 medium containing 20% ​​inactivated fetal bovine serum, 100 U / ml penicillin, and 100 μg / mL streptomycin in a 37°C, 5% CO2 incubator. The cells were passaged every 4 to 5 days after confluence. Tumor cells in the logarithmic growth phase were used for inoculation of tumors in vivo.

[0817] OVCAR3 tumor cells were adjusted to a concentration of 1×10 8 / mL, inoculated subcutaneously in the right flank of M-NSG experimental mice, 100uL / mouse, and the tumor cell inoculation volume was 1×10 7 / mice.

[0818] When the tumor grows to about 150 mm 3 The control group was given PBS, and the treatment group was given the anti-human folate receptor (FRa) humanized monoclonal antibody Farletuzumab-LD-38 antibody drug conjugate. The dosage was 5 mg / kg, a single dose. The tumor size was measured twice a week, and the tumor volume (TV, mm 3 ), and draw the tumor growth curve.

[0819] TV(mm 3 )=(L×W 2 ) / 2, L represents the tumor length, W represents the tumor width

[0820] Tumor inhibition rate (%) = (TV control group - TV administration group) × 100 / TV control group

[0821] Mouse tumor curves were plotted using GraphPad Prism software (GraphPad Software, San Diego, California) in XY mode, with the number of days after the first dose as the horizontal axis and tumor volume as the vertical axis. Two-way ANOVA was used to analyze statistical differences between each group and the vehicle control PBS group: *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. The results are shown in Figure 13.

[0822] Summary: As shown in Figure 13, one week after a single dose of 5 mg / kg, the Farletuzumab-LD-38 antibody drug conjugate exhibited tumor growth inhibition compared to the control group, and tumor shrinkage subsequently occurred. On day 24, the tumor inhibition rate (TGI) reached 93.5%, which was significantly different from the control group.

[0823] 4. Folate receptor-positive IGROV1 cell mouse transplant tumor model

[0824] IGROV1 human ovarian cancer tumor cells expressing the folate receptor (FRa) were cultured in DMEM medium containing inactivated 10% fetal bovine serum, 100 U / ml penicillin, and 100 μg / ml streptomycin at 37°C in a 5% CO2 incubator. The cells were passaged every 3 to 4 days after confluence. Tumor cells in the logarithmic growth phase were used for inoculation of tumors in vivo.

[0825] IGROV1 tumor cells were adjusted to a concentration of 1×10 8 / ml, inoculated subcutaneously in the right flank of BALB / c nude experimental mice, 100 μL / mouse, and the tumor cell inoculation volume was 1×10 7 / mice.

[0826] When the average tumor volume reaches 150 mm 3 The animals were divided into two groups (the day of grouping was designated as D0) and each model was divided into two groups, with 5 animals in each group. The control group was given PBS, and the treatment group was given the anti-human folate receptor (FRa) humanized monoclonal antibody Farletuzumab-LD-38 antibody drug conjugate. The dosage was 5 mg / kg, once a week for 2 times. The tumor size was measured twice a week and the tumor volume (TV, mm 3 ), and draw the tumor growth curve.

[0827] TV(mm 3 )=(L×W 2 ) / 2, L represents the tumor length, W represents the tumor width

[0828] Tumor inhibition rate (%) = (TV control group - TV administration group) × 100 / TV control group

[0829] GraphPad Prism software (GraphPad Software, San Diego, California) was used to plot mouse tumor curves in XY mode, with the number of days after the first dose as the horizontal axis and the tumor volume as the vertical axis. Two-way ANOVA was used to analyze the statistical differences between each group and the vehicle control PBS group: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

[0830] The results are shown in Figure 14. After two doses of 5 mg / kg once a week, the Farletuzumab-LD-38 antibody drug conjugate showed rapid tumor shrinkage relative to the control group, with a tumor inhibition rate (TGI) of 100% on day 30, and the efficacy was maintained 50 days after administration.

[0831] Test Example 22 Pharmacokinetics in rats

[0832] The experiment used 3 male SD rats and randomly divided them into 3 groups. They were given a single intravenous injection of 9D1-LD-38 prepared in the test example and 9D1-GGFG-Exatecan prepared in the comparative example, with a dose of 10 mg / kg. Whole blood was collected from the animals in each test group before administration and 5 minutes, 1 hour, 5 hours, 24 hours (D2), 48 hours (D3), 120 hours (D6), 168 hours (D8), 240 hours (D11), 336 hours (D15), and 504 hours (D22) after the start of administration, and serum and plasma were separated. The concentrations of Total Ab (total antibody) and ADC (bound antibody) of each test sample in the serum samples were analyzed by ELISA. The pharmacokinetic parameters of each animal were calculated using the PK Solver non-compartmental model (NCA).

[0833] Determination of total antibody and bound antibody: ELISA method was used to determine the blood concentration of total antibody and bound antibody. The specific method is as follows:

[0834] a) Dilute anti-human IgG (Fab) 2 to 1 μg / mL in PBS, coat a 96-well ELISA plate with 100 μL / well, and incubate at 4°C overnight.

[0835] b) Wash the plate three times and block with 3% BSA-PBST (300 μL / well) at room temperature for 2 h.

[0836] c) Wash the plate three times and dilute 9D1-LD-38 / 9D1-GGFG-Exatecan in rat blank serum at a 2-fold dilution ratio, starting at 1 μg / mL, for a total of seven concentrations. Set the "0" point as rat blank serum. Simultaneously, dilute samples at different post-drug time points with rat blank serum at appropriate dilution ratios. Dilute all samples 50-fold (9D1-GGFG-Dxd) and 100-fold (9D1-LD-38) in 1% BSA-PBST. Add 100 μL / well to the plate and incubate at room temperature for 2 hours.

[0837] d) Wash the plate 6 times.

[0838] Detect total antibody: dilute HRP-goat anti-human IgGFC at 1:20,000 with 1% BSA-PBST, add 100 μL / well to the ELISA plate, and incubate at room temperature for 1 hour.

[0839] ADC measurement: Dilute anti-exatecan antibody to 0.5 μg / mL in 1% BSA-PBST, add 100 μL / well to the ELISA plate, and incubate at room temperature for 1 hour. Wash the plate six times, then dilute HRP-goat anti-mouse IgG (H+L) at a 1:20,000 dilution in 1% BSA-PBST, add 100 μL / well to the ELISA plate, and incubate at room temperature for 1 hour.

[0840] Wash the plate six times, add TMB to the ELISA plate at 100 μL / well, and develop the color at room temperature in the dark for approximately 10 minutes.

[0841] 50 μL / well of stop solution was added and the results were detected on a microplate reader (wavelength 450 / 630 nm).

[0842] Table 8 shows the pharmacokinetic parameters of 9D1-LD-38 and 9D1-GGFG-Exatecan in rats

[0843] Summary: As shown in Table 8 and Figure 15, after a single intravenous injection of 10 mg / kg to rats, the serum drug concentrations of total antibody and bound antibody (ADC) of 9D1-GGFG-Exatecan decreased rapidly (Figure 15), and the half-life was short, 65.1 hours and 57.8 hours, respectively. The AUC ratio of bound antibody to total antibody was 0.67.

[0844] The serum drug concentrations of 9D1-LD-38 total antibody and bound antibody decreased relatively slowly ( FIG15 ), with half-lives of 145.4 h and 119.8 h, respectively, and the AUC ratios of bound antibody and total antibody were 0.78.

[0845] The half-life of 9D1-LD-38-bound antibody was twice that of 9D1-GGFG-Exatecan; the AUC of 9D1-LD-38-bound antibody was 1.3 times that of 9D1-GGFG-Exatecan; the ratio of AUC of 9D1-LD-38-bound antibody to total antibody was greater than that of 9D1-GGFG-Exatecan (0.78 vs 0.67), indicating that 9D1-LD-38 was more stable and the small molecule was not easily shed.

[0846] In summary, the drug clearance of 9D1-LD-38 in rats, whose linker contains an internal salt water-solubilizing group, is significantly weaker than that of 9D1-GGFG-Exatecan, whose linker does not contain a water-solubilizing group, and its in vivo stability is better, as manifested by a significantly prolonged half-life of 9D1-LD-38, a significantly increased drug exposure, and a higher AUC ratio of bound antibody to total antibody.

[0847] Test Example 23 (In vitro cell killing effects of 9D1-LD-38, 9D1-LD-82, 9D1-LD-88, and 9D1-LD-89)

[0848] MC38-hClaudin18.2 cells were cultured in RPMI 1640 (Thermo, Cat: 11875-085) supplemented with 10% FBS (Thermo, Cat: 10099-141C) and 1 μg / mL puromycin (Thermo, Cat: A11138-03) in a 37°C, 5% CO2 incubator for adherent culture. When the cell confluence was greater than 80%, the cells were passaged at a ratio of 1:10 to 1:20 and every 3 days. MC38-hClaudin18.2 cells were digested with recombinant enzyme digestion solution (TrypLE™ Express Enzyme (Chinese name: trypsin), Thermo, Cat: 12605028). After counting, an appropriate amount of cells were resuspended in RPMI 1640 + 10% FBS to adjust the cell density to 1.5×10 4 Cells were seeded into 96-well cell culture plates at a concentration of 100 μL / well and incubated overnight in a 37°C, 5% CO2 incubator. The next day, 9D1-LD-38, 9D1-LD-82, 9D1-LD-88, and 9D1-LD-89 were serially diluted 10-fold starting at 500 nM, for a total of eight dilutions. A negative control with a sample concentration of 0 and a blank control with the same volume of culture medium (200 μL / well) without cells were also set up. The diluted samples were added to the 96-well cell culture plates containing cells at a concentration of 100 μL / well (in duplicate) and incubated in a 37°C, 5% CO2 incubator for 72 hours. Add 20 μL of AlamarBlue (Thermo, Cat: DAL1100) detection reagent to each well and incubate at 37°C for 6 hours. Fluorescence signal intensity is measured on a multi-function microplate reader (Molecular Devices, M5) at an excitation wavelength of 560 nm and an emission wavelength of 590 nm. Assuming the cell viability of the negative control is 100%, calculate the cell viability (%) of the experimental wells according to the following formula: Cell viability (%) = (OD value of the experimental well - OD value of the blank well) / (OD value of the negative control well - OD value of the blank control well) × 100%

[0849] The logarithm of ADC concentration was used as the abscissa, and the cell viability was used as the ordinate. Sigmoidal dose-response (Variable Slope) method (GraphPad Prism software, GraphPad Software, San Diego, California) was used for nonlinear regression to obtain a cell proliferation inhibition curve, as shown in Figure 16.

[0850] Summary: As shown in Figure 16, 9D1-LD-38, 9D1-LD-82, 9D1-LD-88 and 9D1-LD-89 can inhibit the proliferation of MC38-hClaudin18.2 cells in a dose-dependent manner. The inhibitory effects of 9D1-LD-38, 9D1-LD-88 and 9D1-LD-89 are comparable (IC50 values ​​are 0.632, 1.458 and 0.757 nM, respectively), while 9D1-LD-82 is slightly weaker (IC50 value is 8.013 nM).

[0851] Test Example 24 (Inducing Immunogenic Cell Death in B16F10-hClaudin18.2 Cells)

[0852] B16F10-hClaudin18.2 cells were revived and cultured in DMEM (Thermo, cat: 10569044) + 10% FBS (Thermo, cat: 10099-141C) + 0.5 μg / mL Puromycin in a 37°C, 5% CO2 incubator for adherent culture. When the cell confluence was greater than 80%, the cell ratio was subcultured at a ratio of 1:10 to 1:20 and every 3 to 4 days.

[0853] When the cells reached a confluence of about 80%, they were digested with recombinant enzyme digestion solution (TrypLE™ Express Enzyme, Thermo, cat: 12605028), resuspended in DMEM + 10% FBS, and an appropriate volume was taken for counting. The cell density was adjusted to 1×10 4Cells were plated at 100 μL / well in a 96-well plate and incubated overnight at 37°C in a 5% CO2 incubator. The next day, the ADC drug was diluted to a final concentration of 750 nM in DMEM + 10% FBS, and the small molecule toxin exatecan was diluted to a final concentration of 10 nM. A blank control with a sample concentration of 0 was set up. The diluted samples were added to the 96-well plate containing cells at 100 μL / well and incubated in a 37°C, 5% CO2 incubator for 48 hours. The 96-well plate was removed from the incubator, digested with Accutase (Thermo, cat: 00-4555-56), and washed once with 1% FBS-PBS. Anti-Calreticulin-AlexaFluor 647 antibody (Abcam, cat: AB196159, diluted 1:100 in 1% FBS-PBS) was added at 100 μL / well and incubated at room temperature in the dark for 30 minutes. After washing twice with 1% FBS PBS, 2 μl of PI dye (Thermo, cat: 00-4555-56) was added to each sample and incubated at room temperature in the dark for 10 minutes. FACS analysis (Bechman, CytoFLEX) was then performed. The ratio of calreticulin (+) to PI (-) cells was plotted.

[0854] Summary: Figure 17 shows that at an ADC concentration of 750 nM, the 9D1-LD-38 / 9D1-GGFG-Dxd-treated group showed a significant increase in the calreticulin(+) / PI(-) cell ratio compared to the control ADC, with the 9D1-LD-38 group showing a higher ratio than the 9D1-GGFG-Dxd group. When the small molecule drug Dxd was loaded at a concentration of 10 nM, the calreticulin(+) / PI(-) cell ratio also increased significantly, with Exatecan (MCE, cat: HY-13631A) exceeding Dxd (MCE, cat: HY-13631D). These results demonstrate that 9D1-LD-38 induces more potent immunogenic cell death than 9D1-GGFG-Dxd, providing a theoretical basis for its combination with immune checkpoint inhibitors.

[0855] Test Example 25 (Byronocytic Effect on HEK293-hClaudin18.2 / HEK293 Cells)

[0856] AlarmaBlue detection method: hClaudin18.2-positive HEK293-hClaudin18.2 cells and hClaudin18.2-negative HEK293 cells were revived and cultured in DMEM + 10% FBS + 250 μg / mL G418 and DMEM + 10% FBS, respectively. The cells were cultured in a 37°C, 5% CO2 incubator for adherence. When the cell confluence was greater than 80%, the cells were passaged at a ratio of 1:10 to 1:20 and every 3 to 4 days.

[0857] When the cells grew to a confluence of about 80%, negative and positive cells were digested with recombinant enzyme digestion solution (TrypLETM Express Enzyme). After counting, an appropriate amount of cell suspension was centrifuged and resuspended in DMEM + 10% FBS to adjust the cell density to 5 × 10 3 Each positive and negative cell group was seeded at 100 μL / well into a 96-well cell culture plate. A mixed group of positive and negative cells was mixed at a 1:1 ratio and seeded at 200 μL / well into a 96-well cell culture plate. The plates were incubated overnight at 37°C in a 5% CO2 incubator. The following day, the ADC drug was diluted to a final concentration of 0.5 nM in DMEM + 10% FBS, and a blank control with a sample concentration of 0 was set up. 100 μL of medium was discarded from each well of the mixed positive and negative cell groups, and the diluted ADC drug was added to the cells at 100 μL / well into the 96-well cell culture plate. The plates were incubated at 37°C in a 5% CO2 incubator for 120 hours. The 96-well cell culture plate was removed from the incubator, 20 μL of AlamarBlue (Thermo, cat: DAL1100) detection reagent was added to each well, and the cells were incubated at 37°C for 4 h. The fluorescence signal intensity was detected on a multifunctional microplate reader at an excitation wavelength of 560 nm and an emission wavelength of 590 nm.

[0858] The cell viability of the blank control was set as 100%, and the cell viability (%) of the experimental wells was calculated according to the following formula:

[0859] Cell viability (%) = (fluorescence value of experimental well / fluorescence value of blank control well) × 100%

[0860] FACS detection method: HEK293 cells were labeled and counted in the logarithmic growth phase. An appropriate amount of cell suspension was centrifuged and washed twice with PBS. The density was then adjusted to 5 × 10 6 / mL, add an appropriate volume of CFSE (eBioscience, 65-0850-84) to a final concentration of 0.5nM, mix immediately, incubate at room temperature in the dark for 10 minutes, add 4-5 times the volume of pre-cooled DMEM + 10% FBS, incubate on ice for 5 minutes to terminate the reaction. After washing twice with pre-cooled DMEM + 10% FBS, adjust the cell density to 2×10 with DMEM + 10% FBS medium. 4 pieces / mL.

[0861] Co-incubation and detection: HEK293-hClaudin18.2 positive cells were digested with recombinant enzyme digestion solution (TrypLETM Express Enzyme). After counting, an appropriate amount of cell suspension was centrifuged and resuspended in DMEM + 10% FBS to adjust the cell density to 2×10 4 Equal volumes of CFSE-labeled HEK293 cells and HEK293-hClaudin18.2 cells were mixed and seeded into 96-well cell culture plates at 100 μL / well. The cells were cultured overnight at 37°C in a 5% CO2 incubator. The next day, the ADC drug was diluted to a final concentration of 0.5 nM in DMEM + 10% FBS medium. A blank control with a sample concentration of 0 was set up. The diluted sample was added to the 96-well cell culture plate containing the cells at 100 μL / well and incubated at 37°C in a 5% CO2 incubator for 72 hours. The supernatant of the 96-well cell culture plate was discarded, and the cells were digested with recombinase digestion solution and washed once with 1% FBS PBS. Each sample was resuspended in 250 μL of 1% FBS PBS and counted to calculate the total cell number of each sample. 100 μL of each sample was taken for FACS analysis to determine the ratio of hClaudin18.2-positive and -negative cells. Based on the total number of cells, the number of hClaudin18.2-positive and -negative cells in each sample was calculated.

[0862] Summary: After 5 days of co-culture of ADC drugs with HEK293, HEK293-hClaudin18.2 and HEK293+HEK293-hClaudin18.2 mixed cultures, the cell viability was detected by the AlarmaBlue method. The results are shown in Figure 18, which includes Figure 18A and Figure 18B, where Figure 18A is the AlarmaBlue method and Figure 18B is the flow cytometry method. As shown in Figure 18A, none of the four ADCs had a killing effect on negative HEK293 cells. Compared with the control (Isotype IgG1) ADC, 9D1-LD-38 and 9D1-GGFG-Dxd both had a killing effect on positive HEK293-hClaudin18.2 cells, and the killing effects of the two were comparable. Compared with the control (Isotype IgG1) ADC, 9D1-LD-38 and 9D1-GGFG-Dxd had a killing effect on both positive and negative mixed cells, and the killing effect of 9D1-LD38 was stronger than that of 9D1-GGFG-Dxd. These results indicate that 9D1-LD-38 produces a stronger bystander killing effect than 9D1-GGFG-Dxd.

[0863] The ADC drug was co-cultured with HEK293 (CFSE-labeled) + HEK293-hClaudin18.2 cells for 3 days. The total number and ratio of positive and negative cells were measured using a cell counter and flow cytometry, respectively. As shown in Figure 18B, the isotype control (Isotype IgG1) ADC had no killing effect on either the positive HEK293-hClaudin18.2 cells or the negative HEK293 cells, while 9D1-LD-38 and 9D1-GGFG-Dxd had killing effects on both HEK293-hClaudin18.2 and negative HEK293 cells, with 9D1-LD38 having a stronger killing effect on negative HEK293 cells than 9D1-GGFG-Dxd. These results indicate that 9D1-LD38 produces a stronger bystander killing effect than 9D1-GGFG-Dxd.

[0864] Test Example 26 (In vitro enzymatic release of free toxin molecules)

[0865] Preparation of activation buffer solution (10 mM L-cysteine ​​(ACRO, cat: 173600250), 15 mM EDTA, 0.1 M His-HCl, pH 5.5): Weigh 3.63 g of solid cysteine ​​and dissolve it in 3 mL of 0.1 M His-HCl, pH 5.5 solution. Then add 90 μL of 0.5 M EDTA solution and mix well.

[0866] Sample Preparation: 1) Add 5 μL of Cathepsin B enzyme solution (0.15 mg / mL, Sino Biological, cat: 10483-H08H) to 30 μL of activation buffer solution, mix well, and incubate in a 37°C water bath for 15 minutes. Then, add 300 μL of 0.1 M His-HCl, pH 5.5, and 36 μL of 9D1-LD-38 (375 μg) and incubate in a 37°C water bath overnight. This is used to confirm the presence of free small molecules. 2) Dilute LD-38, LD-65, and Deruxtecan (MC-GGFG-Dxd) to 14 mg / mL with DMSO; take 10 μL of the diluted linker-toxin, add 20 μL of activation buffer to each, then add 250 μL (LD-38), 244.5 μL (LD-65), 344.8 μL (MC-GGFG-Dxd), activation buffer, mix well, and incubate in a 37°C water bath for 15 minutes; take 0.4 mg / mL of Cathepsin B enzyme (Sino Biological, cat: 10483-H08H) was added to 950 μL of activation buffer and incubated at 37°C in a water bath for 15 min. Then, 230 μL of activated linker-toxin solution was added to 230 μL of activated enzyme solution (molar ratio of linker-toxin to Cathepsin B approximately 1200:1) and incubated at 37°C in a water bath for 0.5, 2, 4, and 20 hours. At each time point, 50 μL of the enzyme digestion solution was removed and 2.5 μL of 10% TFA solution was added to terminate the reaction. Kinetic analysis of enzyme digestion was performed, as shown in Figure 19.

[0867] Negative control preparation: Place 30 μL of activation buffer in a 37°C water bath for 15 minutes. Add 300 μL of 0.1 M His-HCl, pH 5.5, and 36 μL of 9D1-LD-38 (375 μg) and incubate in a 37°C water bath overnight. RP-HPLC (Proteomix RP-1000) parameters are shown in Table A:

[0868] Table A

[0869] Summary: As shown in Figure 19, the toxin molecules released from 9D1-LD-38 after Cathepsin B cleavage are consistent with the peak time of the standard Exatecan, and no interference peak appears in the control group (Control) without Cathepsin B enzyme. Therefore, it is determined that Cathepsin B enzyme can effectively cleave the linker of 9D1-LD-38, and the cleavage product is free Exatecan.

[0870] The enzymatic cleavage kinetics of LD-38, LD-65, and Deruxtecan (MC-GGFG-Dxd) are shown in Table 9 below. LD-38 can effectively release toxin molecules after cleavage by Cathepsin B, and the cleavage is almost complete (95%) after 4 hours. LD-65 and MC-GGFG-Dxd are insensitive to the Cathepsin B enzyme. After 20 hours of cleavage, only a small amount of toxin is released from LD-65 (4%), and no cleavage product is released from MC-GGFG-Dxd.

[0871] Table 9 Cathepsin B cleavage kinetics by LD-38, LD-65, and Deruxtecan (MC-GGFG-Dxd)

[0872] Test Example 27 (In vitro serum stability)

[0873] Sample incubation: 9D1-LD-38, 9D1-LD-65, 9D1-LD-73, 9D1-LD-74, and 9D1-Deruxtecan (9D1-GGFG-Dxd), prepared in the aforementioned experimental examples, each with a DAR value of 8, were prepared with human, cynomolgus macaque, and rat serum (9D1-LD-38 and 9D1-LD-65 were supplemented with mouse serum) at a final concentration of 300 μg / mL. 200 μL / tube was plated in triplicate. A negative control (0.5% BSA-PBS) and a standard were also included. All samples were incubated at 37°C for 0, 1, 3, 7, 14, and 21 days, followed by determination of free exatecan.

[0874] Sample preparation: 10.0 μL of standard curve samples, quality control samples, test samples, double blank samples (10.0 μL matrix sample), and zero concentration samples (10.0 μL matrix sample) were added to strip tubes, and 25.0 μL of internal standard working solution (Exatecan-D5 concentration was 3.00 ng / mL, and 25.0 μL of internal standard diluent was added to the double blank samples); 90.0 μL of methanol:acetonitrile (1:1, v / v) was added to each well, and the tubes were shaken at 2500 rpm for 5 min; the tubes were centrifuged at 3800 g at 4°C for 5 min; 80.0 μL of supernatant was collected and dried with nitrogen at 40°C; 100 μL of 0.1% FA in 15% MeOH was added to each well, mixed well, and analyzed by LC-MS.

[0875] Sample analysis: Liquid chromatography (Nexera, Shimadzu) conditions: mobile phase A: 0.1% formic acid in water; mobile phase B: 0.1% acetonitrile solution; column: ACE Excel 3μm 50 x 2.1 mm; flow rate: 0.6 mL / min; column temperature: 40°C; mass spectrometry analysis (Triple Quad 7500, AB / Sciex); data acquisition (Analyst, Version 1.6.2, Applied Biosystems / Sciex).

[0876] Data processing: Through formula C 毒素 = (toxin molecular weight * DAR) / ADC molecular weight * 300 μg / mL to calculate the toxin content in ADC, serum toxin release rate = serum toxin content / C 毒素 A curve was drawn using Graphpad Prism 9 with the incubation time as the horizontal axis and the release rate as the vertical axis.

[0877] Summary: In vitro serum stability studies show exatecan release rate versus time, as shown in Figure 20. Exatecan release from the ADC increases with prolonged incubation time. Figure 20 includes Figures 20A, 20B, 20C, 20D, 20E, and 20F. 9D1-LD-38 (Figure 20A) releases only 2.8%, 0.9%, and 3.3% of the toxin in human, cynomolgus monkey, and rat serum, respectively, after 21 days of incubation at 37°C (peak release in rat serum occurs approximately 7 days after incubation). 9D1-Deruxtecan (9D1-GGFG-Dxd) (Figure 20B) releases 8.5% (human serum), 4.6% (cynomolgus monkey serum), and 5.3% (rat serum), respectively, after 21 days of incubation under the same conditions. This represents 3-fold, 5-fold, and 1.5-fold the toxin release of 9D1-LD-38 in serum from the corresponding species. The serum stability of 9D1-LD-65 (Figure 20C), 9D1-LD-73 (Figure 20D), and 9D1-LD74 (Figure 20E) in humans, cynomolgus macaques, and rats was similar to that of 9D1-LD-38. The maximum toxin release rate in human serum was approximately 3%, while the maximum toxin release rates in cynomolgus macaque and rat serum were both below 1.5%. The toxin release of 9D1-LD-38 in mouse serum was slightly higher than that of 9D1-LD-65 (Figure 20F), but the difference was not significant. The maximum release rate of both agents did not exceed 2% over 21 days.

[0878] Test Example 28 (Dynamic Changes of ADC DAR Values ​​in Rats)

[0879] Three male SD rats were randomly divided into three groups and administered a single intravenous bolus of 10 mg / kg to each of the test articles: 9D1-LD-38, 9D1-LD-65, and 9D1-Deruxtecan (9D1-GGFG-Dxd) (DAR values ​​of 8, respectively) prepared in the aforementioned experimental examples. Whole blood was collected from each test article group before administration and at 5 minutes, 1 hour, 5 hours, 24 hours (D2), 48 hours (D3), 120 hours (D6), 168 hours (D8), 240 hours (D11), 336 hours (D15), and 504 hours (D22) after the start of administration. The blood was divided into two aliquots for serum and plasma collection, respectively, and stored at -60°C.

[0880] Serum total antibody determination method: 10.0 μL of standard curve sample, quality control sample, test sample, double blank sample (10.0 μL matrix sample) and zero concentration sample (10.0 μL matrix sample) were added to a row of tubes; 100 μL of precipitant (1% trichloroacetic acid in isopropanol solution) was added to each well, shaken at 2000 rpm for 5 min, centrifuged at 10°C, 500 g for 5 min, and the supernatant was removed; 100 μL of ammonium bicarbonate solution (100 mM, pH = 8.0) was added to each well, vortexed to mix, the precipitate was resuspended, centrifuged at 10°C, 1000 g for 5 min, and the supernatant was removed; 50.0 μL of ammonium bicarbonate solution (100 mM, pH = 8.0) was added to each well, vortexed to mix; 10.0 μL of Sigma trypsin solution (Sigma, cat: Y0002311) (5 mg / mL), centrifuged at 3500 rpm for 30 s, vortexed at 1000 rpm for 3 min, placed in a shaking incubator at 60°C, 550 rpm, and incubated for 1.5 hours; after the incubation, 20.0 μL of 10% FA in each tube was added. ACN (terminator); for zero-concentration samples, standard curve samples, quality control samples, and test samples, 10.0 μL of internal standard working solution (TTPP-IS, human Fc universal peptide, 5000 ng / ml) was added; for double blank samples, 10.0 μL of internal standard preparation solvent (ammonium bicarbonate solution (100 mM, pH = 8.0)) was added. The above samples were centrifuged at 3800 g for 10 seconds, vortexed at 1200 rpm for 2 minutes; centrifuged at 3800 g for 5 minutes, 50.0 μL of the supernatant was taken, 50.0 μL of water was added to each well, mixed evenly, and analyzed by LC-MS.

[0881] Serum-bound toxin assay method: 10.0 μL of standard curve samples, quality control samples, test samples, double blank samples (10.0 μL matrix sample) and zero concentration samples (10.0 μL matrix sample) were added to a series of tubes; 90 μL of precipitant (1% trichloroacetic acid in isopropanol solution) was added to each well, shaken at 2000 rpm for 5 minutes, centrifuged at 10°C and 500 g for 5 minutes, and the supernatant was removed; 60 μL of ammonium bicarbonate solution (100 mM, pH = 6.0) was added to each well, vortexed to mix, the precipitate was resuspended, centrifuged at 10°C and 1000 g for 5 minutes, and the supernatant was removed; 60.0 μL of ammonium bicarbonate solution (100 mM, pH = 8.0) was added to each well, vortexed to mix; 60.0 μL of papain (Adamas life, cat: 88915D, 5 mg / mL), centrifuged at 3500 rpm for 30 s, vortexed at 1000 rpm for 3 min, placed in a shaking incubator, 40°C, 550 rpm, incubated for 2 hours; 10 μL of 1% formic acid aqueous solution was added to each well, centrifuged at 3500 rpm for 30 s, 40°C, 550 rpm, and incubated for 0.5 hours; for zero concentration samples, standard curve samples, quality control samples, and test samples, 25.0 μL of internal standard working solution ((Exatecan-D5, 1000 ng / ml)) was added; for double blank samples, 25.0 μL of internal standard preparation solvent was added, the above samples were centrifuged at 3800 g for 10 seconds, vortexed at 1200 rpm for 2 min; 200 μL of pre-cooled acetonitrile was added to each well, shaken at 2500 rpm for 5 min, centrifuged at 3800 g for 5 min, 50.0 μL of supernatant was taken, and 150 μL was added to each well. 0.5% formic acid in water, mixed well, and analyzed by LC-MS.

[0882] The total Ab (total antibody) and bound toxin concentrations of each test article in serum samples were analyzed by LC / MS, and the bound antibody DAR values ​​at each time point of the animals were calculated using DAR = bound toxin concentration (mmol / L) / total antibody concentration (mmol / L).

[0883] Summary: The DAR value-time curves of the ADC in the serum of each test article group are shown in Figure 21. Following a single intravenous bolus of 10 mg / kg to rats, the DAR values ​​of 9D1-LD-38, 9D1-LD-65, and 9D1-Deruxtecan (9D1-GGFG-Dxd) gradually decreased over time. The DAR curves for 9D1-LD-65 and 9D1-LD-38 were nearly identical and maintained at relatively high DAR levels, with DAR values ​​remaining around 7 at 48 hours post-dose and remaining around 3-4 at 21 days. The DAR value of 9D1-Deruxtecan (9D1-GGFG-Dxd) decreased more rapidly, dropping from approximately 7.5 at the beginning of dosing to 5.5 at 48 hours and to approximately 1.6 at 21 days. The above results indicate that after in vivo administration, 9D1-LD-38 and 9D1-LD-65 are more stable than 9D1-Deruxtecan (9D1-GGFG-Dxd), as shown by higher DAR.

[0884] Test Example 29 (Tissue Distribution in Tumor-Bearing Mice)

[0885] Tumor-bearing mouse modeling and administration: Male C57BL / 6 mice (6 weeks old) were subcutaneously inoculated with MC38-hClaudin18.2 cells (1 x 10 6 When the tumor volume of the mice grew to ∼300 mm3, the mice were divided into groups and dosed progressively. 9D1-LD-38, 9D1-LD-65, 9D1-Deruxtecan (9D1-GGFG-Dxd), and PBS prepared in the above-mentioned experimental example were injected into the tail vein. The DAR values ​​were 8, and the dose was 10 mg / kg.

[0886] Sample Collection: Blood and tumor tissue were collected 24, 72, and 168 hours after administration, with samples collected from three mice at each time point. Whole blood was collected from tumor-bearing mice via the orbital cavity and placed in an anticoagulant tube containing EDTA-K2. Plasma was then centrifuged within 30 minutes. Mice were sacrificed by CO2 asphyxiation, and tumors were collected. Plasma and tumor tissue were stored at -80°C.

[0887] Determination of free toxin in plasma: 10.0 μL of standard curve samples, quality control samples, test samples, double blank samples (10.0 μL matrix sample), and zero concentration sample (10.0 μL matrix sample) were added to a strip tube, and 25.0 μL of internal standard working solution (Exatecan-D5 concentration was 3.00 ng / mL, and 25.0 μL of internal standard diluent was added to the double blank sample); 90.0 μL of methanol:acetonitrile (1:1, v / v) was added to each well, and the tubes were shaken at 2500 rpm for 5 min; centrifuged at 3800 g at 4°C for 5 min; 80.0 μL of supernatant was taken and dried with nitrogen at 40°C; 100 μL of 0.1% FA in 15% MeOH was added to each well, mixed evenly, and analyzed by LC-MS.

[0888] Determination of total toxins (bound toxins + free toxins) in blood: 10.0 μL of standard curve samples, quality control samples, test samples, double blank samples (10.0 μL matrix samples), and zero concentration samples (10.0 μL matrix samples) were added to strip tubes, 10.0 μL of papain (Adamas life, cat: 88915D, 5 mg / ml) was added to each well, and centrifuged at 3500 rpm for 1 min; the samples were placed in a shaking incubator at 40°C, 550 rpm, and incubated for 6-8 hours; 1.00 μL of 1% FA in each well was added H2O, centrifuge at 3500rpm for 1min, incubate at 40℃, 550rpm for another 30min; add 10.0μL of internal standard working solution (Exatecan-D5, 1000ng / ml) to the zero concentration sample, standard curve sample, quality control sample, and test sample, and add 10.0μL of internal standard preparation solvent to the double blank sample; centrifuge the above samples at 3800g for 10 seconds and vortex at 1200rpm for 2min; add 100μL of pre-cooled ACN to each well, shake at 2500rpm for 5min, and centrifuge at 3500g for 5min; take 30.0μL of supernatant, add 90.0μL of 0.5% FA in H2O to each well, vortex for 3min, and analyze by LCMS.

[0889] Determination of free toxins in tumor tissues: Tumor tissue (drug administration and blank PBS groups) homogenization treatment: Tumor tissue was weighed and added with 10 times the volume of 50% methanol water for homogenization treatment. The homogenate of the blank PBS group was used as the matrix standard curve and quality control sample configuration. 10.0 μL of standard curve samples, quality control samples, test samples, double blank samples (10.0 μL matrix samples) and zero concentration samples (10.0 μL matrix samples) were added to strip tubes, and 25.0 μL of internal standard working solution (Exatecan-D5 concentration was 3.00 ng / mL, and 25.0 μL of internal standard diluent was added to the double blank samples); 90.0 μL of methanol:acetonitrile (1:1, v / v) was added to each well, and the tubes were shaken at 2500 rpm for 5 min; centrifuged at 4°C, 3800 g for 5 min; 80.0 μL of supernatant was taken and dried with nitrogen at 40°C; 100 μL of 0.1% FA in 15% MeOH was added to each well, vortexed for 3 min, and analyzed by LCMS.

[0890] Determination of total toxin (bound toxin + free toxin) in tumor tissue: 10.0 μL of standard curve samples, quality control samples, test samples, double blank samples (10.0 μL matrix sample), and zero concentration samples (10.0 μL matrix sample) were added to a series of tubes; 10.0 μL of papain (Adamas life, cat: 88915D 5 mg / ml) was added to each well and centrifuged at 3500 rpm for 1 min; the tubes were placed in a shaking incubator at 40°C, 550 rpm, and incubated for 6-8 hours; 1.00 μL of 1% FA in each well was added H2O, centrifuge at 3500rpm for 1min, incubate at 40℃, 550rpm for another 30min; 5. For zero-concentration samples, standard curve samples, quality control samples, and test samples, add 10.0μL of internal standard working solution (Exatecan-D5, 1000ng / ml); for double blank samples, add 10.0μL of internal standard preparation solvent; centrifuge the above samples at 3800g for 10 seconds and vortex at 1200rmp for 2min; add 100μL of pre-cooled ACN to each well, shake at 2500rpm for 5min; centrifuge at 3500g for 5min; take 30.0μL of supernatant, add 90.0μL of 0.5% FA in H2O to each well, vortex for 3min, and analyze by LCMS.

[0891] Sample analysis: Liquid chromatography (Nexera, Shimadzu) conditions: mobile phase A: 0.1% formic acid in water; mobile phase B: 0.1% acetonitrile solution; column: ACE Excel 3μm 50x 2.1mm; flow rate: 0.6mL / min; column temperature: 40℃; mass spectrometry analysis (Triple Quad 7500, AB / Sciex); data acquisition (Analyst, Version 1.6.2, Applied Biosystems / Sciex). GraphPad Prism 9 software was used to input the time points of sample collection and the corresponding toxin contents to draw toxin content curves in plasma and tumor tissues. The results are shown in Figures 22 and 23. Figure 22 includes Figures 22A and 22B, wherein Figure 22A shows the total toxin (bound toxin + free toxin) in the plasma of tumor-bearing mice, and Figure 22B shows the free toxin content in the plasma of tumor-bearing mice. Figure 23 includes Figures 23A and 23B, wherein Figure 23A shows the total toxin (bound toxin + free toxin) in the tissues of tumor-bearing mice, and Figure 23B shows the free toxin content in the tissues of tumor-bearing mice.

[0892] Summary: As shown in Figure 22, the free and total toxin levels in the plasma of animals in each test group showed similar trends, reaching a peak at 24 hours and then slowly declining. Total toxin levels in the 9D1-LD-38 and 9D1-LD-65 groups (Figure 22A) were essentially identical, ranging from 3000-4000 ng / mL at 24 hours, and decreased by approximately 70%-80% after 7 days. The total toxin levels in the plasma of 9D1-Deruxtecan (9D1-GGFG-Dxd) were approximately half (1 / 2) those of 9D1-LD-38 and 9D1-LD-65 at all time points. The free toxin levels and trends in the plasma of animals in the 9D1-LD-38, 9D1-LD-65, and 9D1-Deruxtecan (9D1-GGFG-Dxd) groups were also similar (Figure 22B), reaching a peak at 24 hours and then slowly declining. Relative to the total toxin content in plasma, the free toxin content was extremely low, with 24-hour levels below 0.4 ng / mL, representing 0.01%-0.02% of the total toxin content. This was also well below the IC50 value for target cell killing in vitro (3 ng / mL), indicating that the toxins of the three ADC drugs were not easily shed in plasma and were relatively stable.

[0893] As shown in Figure 23, total toxin levels in tumor tissues of all treatment groups reached their highest level 24 hours after administration (Figure 23A), then rapidly declined to approximately one-third of the 24-hour level after 168 hours. The accumulation levels of 9D1-LD38 and 9D1-LD-65 in tumor tissues were significantly higher than those of 9D1-Deruxtecan (9D1-GGFG-Dxd). Free toxin levels in tumor tissues of all treatment groups reached their highest level 24 hours after administration (Figure 23B), gradually decreasing by 72 hours and then remaining largely constant until 168 hours. Free toxin levels in tumor tissues of 9D1-LD-38 and 9D1-LD-65 remained significantly higher than those of 9D1-Deruxtecan (9D1-GGFG-Dxd) throughout the entire time period (2-5 times), with 9D1-LD-38 achieving the highest level. The results of the total toxin (bound toxin + free toxin) and free toxin content in the tumor tissue of the above-mentioned tumor-bearing mice showed that 9D1-LD-38 and 9D1-LD-65 had better penetration and enrichment in the tumor tissue, and released higher levels of free toxins, thus being able to produce better tumor treatment effects compared with 9D1-Deruxtecan (9D1-GGFG-Dxd).

[0894] Test Example 30 (Efficacy in a P-gp-mediated drug-resistant tumor-bearing mouse model)

[0895] Tumor-bearing mouse modeling and drug administration: BALB / c Nude mice (female, 5-6 weeks old) were subcutaneously inoculated with MC38hClaudin18.2 cells (1×10 6 When the tumor volume grew to about 120 mm3, the patients were randomly divided into groups and given medication according to Table 10. The tumor volume and body weight were measured twice a week and the tumor volume (TV, mm 3 ), draw the tumor growth curve. TV (mm 3 )=(L×W 2 ) / 2, where L represents tumor length and W represents tumor width. GraphPad Prism software (GraphPad Software, San Diego, California) was used to plot mouse tumor growth and body weight curves in XY mode. Tumor volume and body weight are expressed as mean ± standard error. The results are shown in Figure 24 , which includes Figures 24A and 24B , with Figure 24A showing the mouse tumor growth curve and Figure 24B showing the mouse body weight curve.

[0896] Table 10 Grouping and dosing regimen of the p-gp-mediated drug-resistant tumor-bearing mouse model Note: ip, Intraperitoneal injection; po, Peros, oral administration; QW, once a week; QD, once a day; 5 on + 2 off / cycle, 7 days a cycle, 5 days of administration and 2 days of rest; 9D1-LD-38 and 9D1-Deruxtecan (9D1-GGFG-Dxd), DAR = 8; 9D1-vd-PAB-MMAE, DAR = 4; Tariquidar, p-gp inhibitor (Cat: HY-10550 / CS-0722 Lot: 242595).

[0897] Summary: P-glycoprotein (P-gp) transmembrane transporter causes chemotherapy resistance through active drug efflux, which is one of the main reasons for treatment failure. The mouse MC38 rectal cancer model has high expression of p-gp and is insensitive to many chemotherapy drugs. The tumor therapeutic effect of ADC drugs alone or in combination with the p-gp inhibitor Tariquidar in the MC38-hClaudin18.2 tumor-bearing mouse model is shown in Figure 24A. Tariquidar alone had no tumor inhibitory effect compared with the PBS control group; the tumor in the 9D1-vc-PAB-MMAE group grew rapidly, and the tumor growth curve showed a weak inhibitory effect compared with the PBS control group (the tumor volume was approximately 2600 mm on the 17th day after administration). 3 and 1900mm 3 ), 9D1-vc-PAB-MMAE+Tariquidar can greatly inhibit tumor growth, causing the tumor to shrink to almost disappear; 9D1-Deruxtecan (9D1-GGFG-Dxd) can significantly inhibit tumor growth, and the tumor inhibitory effect of 9D1-Deruxtecan (9D1-GGFG-Dxd)+Tariquidar is slightly enhanced compared with 9D1-Deruxtecan (9D1-GGFG-Dxd) alone, but neither can lead to tumor shrinkage (the tumor volume was approximately 1000mm on the 17th day after administration). 3 and 660mm 3 ); Both 9D1-LD-38 alone and 9D1-LD-38+Tariquidar were able to strongly inhibit tumor growth and cause the tumor to shrink to almost disappearance. The body weight of the mice remained within the normal fluctuation range after administration, and no weight loss was observed due to treatment (Figure 24B). The above results show that compared with 9D1-vc-PAB-MMAE and 9D1-Deruxtecan (9D1-GGFG-Dxd), 9D1-LD-38 can resist tumor resistance caused by p-gp, and single-drug administration can produce a good tumor treatment effect.

[0898] Test Example 31 (Anti-tumor effect of combined ICI administration in tumor-bearing mice)

[0899] Tumor-bearing mouse modeling and drug administration: 4T1-hClaudin18.2 cells were cultured in 1640 complete medium (supplemented with 10% FBS and 2 μg / mL puromycin, Gibco, Cat: A1113803) in adherent flasks in a 37°C, 5% CO2 incubator. When the cells reached approximately 80% confluence, they were digested with TRYPLE EXPRESS trypsin (Gibco, Cat: 12605028) and centrifuged to collect 2.5×10 7 Cells were collected and washed once with PBS. The supernatant was discarded and resuspended in PBS containing 50% Matrigel (Corning, Cat: 326234) in a total volume of 5 ml. 100 μL of cell suspension was inoculated into the right abdominal skin of BALB / c female mice (female, 5-6 weeks old). Tumors grew to approximately 150 mm3 in volume and were randomly divided into groups and dosed according to Table 11. Tumor size was measured twice a week with a vernier caliper. TV (mm 3 )=(L×W 2 ) / 2, L represents tumor length, and W represents tumor width. GraphPad Prism software (GraphPad Software, San Diego, California) was used to plot mouse tumor growth curves in XY mode, and tumor volumes were expressed as mean ± standard error.

[0900] Table 11 Grouping and dosing regimen of ICI combination therapy in tumor-bearing mouse model Note: ip, intraperitoneal injection; QW, once a week; BIW, twice a week; αPD-1, anti-mouse PD-1 antibody (InVivoMAb (anti-mouse PD-1, Cat: BE0146).

[0901] Summary: Immune checkpoint inhibitors (ICIs) have become one of the cornerstones of cancer treatment. The combined therapeutic effect of 9D1-LD-38 and the anti-mouse PD1 drug αPD-1 in the 4T1-hClaudin18.2 tumor-bearing mouse model of immune-competent BALB / c mice is shown in Figure 25. Compared with the PBS control group, both 9D1-LD38 monotherapy and αPD-1 monotherapy were able to partially inhibit tumor growth (tumor volumes were approximately 850 mm and 100 mm, respectively, on day 14 after administration). 3 , 480mm 3 , 426mm3 ), the combined administration of 9D1-LD38+αPD-1 can greatly inhibit tumor growth and even produce significant tumor regression (the tumor volume was about 70mm on the 10th day after administration). 3 ).

[0902] Test Example 32 (Anti-tumor effect of bystander action on tumor-bearing mice)

[0903] Tumor-bearing mouse modeling and drug administration: MC38-hClaudin18.2 cells and MC38-luc (luciferase) were cultured in DMEM medium (Meilunbio, cat: MA0212-Nov-02H) containing inactivated 10% fetal bovine serum (Excell Bio, cat: 12A222) and 1x penicillin / streptomycin (Solarbio, cat: 20221231) at 37°C in a 5% CO2 incubator. Tumor cells in the logarithmic growth phase were used for inoculation of tumors in vivo. MC38-hClaudin18.2 cells and MC38-luc cells were mixed (quantity ratio 1:1) and resuspended in PBS to adjust the cell concentration to 5×10 6 / ml, inoculated subcutaneously on the right side of the abdomen of BALB / c nude mice, 100μL / mouse. 3 Around 8:00 am, the mice were divided into groups and given the drug (this day was designated as PG-D0). There were 4 groups in total, with 5 mice in each group. The specific dosing schedule is shown in the table below:

[0904] Table 12 Grouping and dosing regimen of bystander effect in tumor-bearing mouse model

[0905] Note: *, the administration volume is 10 μl / g body weight;

[0906] iv: intravenous injection; QW×2: administered once a week for a total of 2 times.

[0907] Tumor volume was measured 2-3 times weekly using a vernier caliper. The long and short diameters of the tumors were measured. The volume was calculated using the formula: volume (TV) = 0.5 × long diameter × short diameter × short diameter. Mice were intraperitoneally injected with luciferin substrate (15 mg / ml, 10 μl / g) and anesthetized with isoflurane. Luminescence signals were collected using a small animal in vivo imager (IVIS Lumina Series III, PerkinElmer).

[0908] Summary: Bystander killing is the ability of ADC drugs to release toxins after being internalized by target-positive tumor cells, and to kill adjacent target-negative tumor cells through transmembrane action. It is considered to be one of the reasons why ADCs can produce good efficacy against tumors with heterogeneous target expression. In this experimental example, target-positive (MC38-hClaudin18.2) cells and target-negative (MC38-luc) cells were mixed and inoculated into mice to produce a tumor-bearing mouse model with heterogeneous target expression. The bystander killing effect of the ADC drug was verified and tested. The results are shown in Figure 26, which includes Figure 26A and Figure 26B. Figure 26A shows the changes in tumor volume of mice after drug administration, and Figure 26B shows the bioluminescent signal of target-negative tumor cells in tumor-bearing mice 3 days and 7 days after drug administration. As shown in Figure 26A, compared with the PBS control group (Contral), ISO (Isotype)-LD-38 grew faster and only produced a weak tumor inhibitory effect (16 days after tumor inoculation, the tumor volume was approximately 2700mm and 2700mm, respectively). 3 and 1700mm 3 ); 9D1-Deruxtecan (9D1-GGFG-Dxd) was able to partially inhibit tumor growth (16 days after tumor inoculation, the tumor volume was about 1000mm 3 ), but the tumor volume continued to increase; 9D1-LD-38 was able to significantly inhibit tumor growth, and by day 16 the tumor was almost completely eliminated, indicating that 9D1-LD-38 was able to kill not only target-positive MC38-hClaudin18.2 cells, but also target-negative MC38-luc cells. The bioluminescent signals of target-negative tumor cells in tumor-bearing mice collected on day 3 (PG-3, day 3 after inoculation) and day 7 (PG-7, day 12 after inoculation) after grouping (Figure 26B) also clearly showed that compared with the control group and the ISO-LD38 and 9D1-Deruxtecan (9D1-GGFG-Dxd) treatment groups, 9D1-LD38 was able to effectively kill target-negative tumor cells and caused the bioluminescent signal to be greatly weakened or eliminated.

[0909] Although the embodiments of the present application are described above in conjunction with the accompanying drawings, the present application is not limited to the above-mentioned specific embodiments and application fields. The above-mentioned specific embodiments are merely illustrative and instructive, and not restrictive. A person of ordinary skill in the art, guided by this specification and without departing from the scope of protection of the claims of this application, may also devise various forms, all of which fall within the scope of protection of this application.

Claims

1. A compound represented by formula (1), or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein: The structure of formula (1) is shown below: M is a linking group; Sp is -L a -Q a -; A is selected from the group consisting of absent, hydrogen, and alkyl; T represents a linear or branched (C1-C8) trivalent alkyl group; Y, Y', and Y" are -L b -Q b -; L a , L b are respectively selected from absence, alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene, heteroarylene or polyethylene glycol group, wherein the alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene, heteroarylene, polyethylene glycol group is optionally substituted with a substituent; Q a , Q b are respectively selected from absent, -C(=O)NR-, -C(=O)O-, -OC(=O)NR-, -S(=O)2NR-, -NRC(=O)-, -OC(=O)-, -NRC(=O)O-, -NRS(=O)2-, -NRC(=O)NR-, -NHC-(=NH)NH-, -C(=O)-, -OC(=O)O-, -O-, -NR-, -S-, S(=O)2-, S(=O)-, or -Se-, wherein R is selected from hydrogen or optionally substituted alkyl; Z is selected from absent, hydrogen, optionally substituted alkyl or sulfonate; Z', Z" are selected from absent, hydrogen, optionally substituted alkyl, sulfonate or phosphate; At least one of Z, Z', and Z" is a sulfonic acid inner salt or a phosphate inner salt; X contains a self-destruct unit or does not exist; D includes drug molecules; i is an integer greater than or equal to 1; h is an integer from 1 to 6; o, p, and q are integers from 0 to 5 respectively.

2. The compound according to claim 1, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein: The formula (1) is formula (1-1): Wherein, the definitions of M, Sp, A, T, Y, h, o, i, X, D, and Z are as described in claim 1; At least one of the i Zs is a sulfonate.

3. The compound according to claim 2, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein: The formula (1-1) is formula (1-1-1): Wherein, AA is an amino acid residue, and W is an integer greater than or equal to 1.

4. The compound according to claim 1, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein: The formula (1) is formula (1-2): Wherein, the definitions of M, Sp, A, T, Y, Z, o, p, i, h, X, Y', and D are as described in claim 1; and Z' is a sulfonic acid inner salt or a phosphate inner salt.

5. The compound according to claim 4, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein: The formula (1) is formula (1-2-1): Wherein, AA is an amino acid residue, and W is an integer greater than or equal to 1.

6. The compound according to claim 1, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein the formula (1) is formula (1-3): in, The definitions of M, Sp, A, Y, T, Z, o, i, h, q, X, Y", and D are as described in claim 1; Z" is a sulfonic acid inner salt or a phosphate inner salt.

7. The compound according to claim 6, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein: The formula (1) is formula (1-3-1): Wherein, AA is an amino acid residue, and W is an integer greater than or equal to 1.

8. A compound according to any one of claims 1 to 7, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: When A and Z do not exist, N, Y, and T form a ring, which is Preferably 9. A compound according to any one of claims 3, 5 or 7, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The amino acids are selected from one or more of alanine, arginine, asparagine, aspartic acid, γ-carboxyglutamate, citrulline, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, norleucine, norvaline, ornithine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine.

10. A compound according to any one of claims 3, 5 or 7, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The amino acids are selected from one or more of valine, glutamic acid, citrulline, lysine, alanine, phenylalanine, arginine, glutamine, asparagine and glycine.

11. A compound according to any one of claims 3, 5 or 7, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The amino acid is selected from one or more of valine, citrulline, alanine, phenylalanine, glutamine and glycine.

12. A compound according to any one of claims 1 to 11, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: M is selected from N3, SH, ONH2, NH2, CO2H, (alkyl)-CO-, HCO-, BrCH 2- 、ICH2-、 13. A compound according to any one of claims 1 to 12, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: M is 14. A compound according to any one of claims 1 to 13, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: M is 15. A compound according to any one of claims 1 to 14, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: A is hydrogen, methyl, ethyl or propyl.

16. A compound according to any one of claims 1 to 15, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: A is hydrogen.

17. A compound according to any one of claims 1 to 16, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The T is one of the following structures:

18. A compound according to any one of claims 1 to 17, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The T is 19. A compound according to any one of claims 1 to 18, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: L a One, two or more selected from alkylene, alkenylene, alkynylene, saturated or unsaturated alicyclic group having 3-10 ring carbon atoms, saturated or unsaturated heteroalicyclic group having 3-10 ring carbon atoms and 1 to 4 heteroatoms, arylene group having 6 to 18 carbon atoms in one or more rings, or heteroarylene group having 6 to 18 carbon atoms in one or more rings and 1 to 4 heteroatoms, or polyethylene glycol group; the alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene group and heteroarylene group are optionally substituted by a substituent, and the heteroatom is selected from nitrogen, oxygen or sulfur.

20. A compound according to any one of claims 1 to 19, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: L a Selected from C1-C 10 Straight or branched alkylene, C2-C 12 Straight or branched alkenylene, C2-C 12 One, two or more of a straight or branched alkynylene group, a saturated or unsaturated alicyclic group having 3-6 ring carbon atoms, a saturated or unsaturated heteroalicyclic group having 3-6 ring carbon atoms and 1 to 4 heteroatoms, or an arylene group having 6 to 10 carbon atoms in one or more rings, a heteroarylene group having 6 to 10 carbon atoms in one or more rings and 1 to 4 heteroatoms, or a polyethylene glycol group, wherein the alkylene group, alkenylene group, alkynylene group, alicyclic group, heteroalicyclic group, arylene group and heteroarylene group are optionally substituted by a substituent, and the heteroatom is selected from nitrogen, oxygen or sulfur.

21. A compound according to any one of claims 1 to 20, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: L a One, two or more selected from C1-C6 straight chain or branched alkylene, C2-C6 straight chain or branched alkenylene, C2-C6 straight chain or branched alkynylene, or polyethylene glycol group, wherein the alkylene, alkenylene and alkynylene are optionally substituted by a substituent.

22. A compound according to any one of claims 1 to 21, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: L a It is one, two or more than three of C1-C6 straight or branched alkylene groups or polyethylene glycol groups, wherein the polyethylene glycol group is -(CH2CH2O) n -、-O(CH2CH2O) n -、-(CH2CH2O) n CH2-、-(CH2CH2O) n CH2CH2-、-CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n CH2-, n is an integer greater than or equal to 1.

23. A compound according to any one of claims 1 to 22, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: L a is ethylene, straight-chain propylene, straight-chain butylene, straight-chain pentylene, or -(CH2CH2O) n One, two or more of CH2CH2-, wherein n is 2, 3, 4, 5, 6, 7 or 8.

24. A compound according to any one of claims 1 to 23, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: L a It is ethylene and / or -(CH2CH2O)4CH2CH2- or pentylene.

25. A compound according to any one of claims 1 to 24, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: L b One or two or more selected from alkylene, alkenylene, alkynylene, saturated or unsaturated alicyclic group having 3-10 ring carbon atoms, saturated or unsaturated heteroalicyclic group having 3-10 ring carbon atoms and 1 to 4 heteroatoms, arylene group having 6 to 18 carbon atoms in one or more rings, or heteroarylene group having 6 to 18 carbon atoms in one or more rings and 1 to 4 heteroatoms, or polyethylene glycol, wherein the alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene group and heteroarylene group are optionally substituted by a substituent, and the heteroatom is selected from nitrogen, oxygen or sulfur.

26. A compound according to any one of claims 1 to 25, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: L b Selected from C1-C 10 Straight or branched alkylene, C2-C 12 Straight or branched alkenylene, C2-C 12 One or two or more of a straight or branched alkynylene group, a saturated or unsaturated alicyclic group having 3-6 ring carbon atoms, a saturated or unsaturated heteroalicyclic group having 3-6 ring carbon atoms and 1 to 4 heteroatoms, or an arylene group having 6 to 10 carbon atoms in one or more rings, a heteroarylene group having 6 to 10 carbon atoms in one or more rings and 1 to 4 heteroatoms, or polyethylene glycol, wherein the alkylene group, alkenylene group, alkynylene group, alicyclic group, heteroalicyclic group, arylene group and heteroarylene group are optionally substituted by a substituent, and the heteroatom is selected from nitrogen, oxygen or sulfur.

27. A compound according to any one of claims 1 to 26, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: L b is selected from C1-C6 straight chain or branched alkylene, C2-C6 straight chain or branched alkenylene, or C2-C6 straight chain or branched alkynylene, or polyethylene glycol group, wherein the polyethylene glycol group is -(CH2CH2O) n -、-O(CH2CH2O) n -、-(CH2CH2O) n CH2-、-(CH2CH2O) n CH2CH2-、-CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n One or two or more than three of CH2-, n is an integer greater than or equal to 1, and the alkylene, alkenylene and alkynylene groups are optionally substituted by a substituent.

28. A compound according to any one of claims 1 to 27, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: L b is a C1-C6 straight or branched chain alkylene group, or a C6-C 10 One or two or more than three of the arylene groups.

29. A compound according to any one of claims 1 to 28, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: L b It is one or two or more of methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, sec-butylene, tert-butylene, or benzylene.

30. A compound according to any one of claims 1 to 29, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: Q a One, two or more selected from -C(=O)NR-, -NR-, -C(=O)O-, -OC(=O)NR-, -NRC(=O)-, -OC(=O)-, -NRC(=O)O-, -NRC(=O)NR-, NHC-(=NH)NH-, -C(=O)-, -OC(=O)O-, wherein R is selected from hydrogen, optionally substituted C1-C6 straight chain or branched alkyl.

31. The compound according to claim 30, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein: Q a It is one or two or more of -C(=O)-, -NR-, -C(=O)NR-, -NRC(=O)NR-, -OC(=O)O-, -OC(=O)NR-, -NRC(=O)- or -OC(=O)-, wherein R is selected from hydrogen or an optionally substituted C1-C6 straight chain or branched alkyl.

32. The compound according to claim 30, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein: Q a It is one or two or more of -C(=O)-, -NR-, -NRC(=O)- and -(C=O)NR-, wherein R is selected from hydrogen or an optionally substituted C1-C6 straight or branched alkyl group.

33. The compound according to claim 30, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein: Q a It is -C(=O)-, -N(CH3)-, -NHC(=O)-, -C(=O)-NH- or -C(=O)-N(CH3)-.

34. A compound according to any one of claims 1 to 33, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: Q b is selected from one, two or more of the group consisting of absence, -C(=O)NR-, -C(=O)O-, -OC(=O)NR-, -NRC(=O)-, -OC(=O)-, -NRC(=O)O-, -NRC(=O)NR-, NHC-(=NH)NH-, -C(=O)-, and -OC(=O)O-, wherein R is selected from hydrogen, an optionally substituted C1-C6 straight chain or branched alkyl.

35. A compound according to claim 34, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: Q b is absent, one or two or more of -C(=O)-, -C(=O)NR-, -OC(=O)NR-, -NRC(=O)-, -NRC(=O)O-, or -NRC(=O)NR-, and R is selected from hydrogen, an optionally substituted C1-C6 straight chain or branched alkyl.

36. A compound according to claim 34, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: Q b is absent, -C(=O)-, -NRC(=O)-, -NRC(=O)NR- or -C(=O)NR-, or two or more thereof; and R is hydrogen or methyl.

37. A compound according to any one of claims 1 to 36, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: X is selected from: The right wavy line in X connects D; R4 is selected from hydrogen, C1-C6 straight or branched alkyl, -(CH2CH2O) m R2, -CH2CH2NR2R3, -CH2CH2SO2R2, R2 and R3 are independently selected from hydrogen, C1-C6 straight chain or branched alkyl, and m is an integer greater than 1.

38. The compound according to claim 37, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein: X is R4 is hydrogen, methyl, -CH2CH2OCH2CH2OH, -CH2CH2NMe2, -CH2CH2SO2Me.

39. The compound according to claim 37, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein: X is R4 is hydrogen or methyl.

40. A compound according to any one of claims 1 to 39, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The sulfonic acid salt is A wavy line connects Y, Y' or Y"; Wherein c, d, e, f, g are all integers of 1, 2, 3, 4, 5 or 6; R6, R7, R 11 , R 12 , R 13 , R 14 are independently selected from absence, hydrogen, and C1-C6 straight or branched chain alkyl.

41. A compound according to claim 40, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The sulfonic acid salt is in, R6, R7, R 11 , R 12 , R 13 and R 14 are respectively selected from absence, methyl or ethyl, c, d, e, f, and g are 1, 2, or 3 respectively.

42. A compound according to claim 40, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The sulfonic acid inner salt is one of the following structures:

43. A compound according to any one of claims 1 to 42, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: The phosphate inner salt is in, a and b are both integers of 1, 2, 3, 4, 5 or 6; R8, R9 and R 10 are selected from hydrogen, C1-C6 straight chain or branched alkyl.

44. A compound according to claim 43, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The phosphate inner salt is in, R8, R9, R 10 They are methyl or ethyl respectively; a is 1 or 2 or 3; b is 1 or 2 or 3.

45. A compound according to claim 43, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The phosphate inner salt is in, R8, R9, R 10 All are methyl; a is 2; b is 1.

46. ​​A compound according to any one of claims 1 to 45, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The D comprises a cytotoxin or an immune agonist.

47. A compound according to claim 46, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The cytotoxin is selected from one or two of a microtubule protein inhibitor, a DNA synthesis inhibitor, a topoisomerase inhibitor or an RNA polymerase 2 inhibitor.

48. A compound according to claim 46, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The cytotoxin is selected from auristatins, maytansine alkaloids, calicheamicin, duocarmycin, pyrrolobenzodiazepines, One or more of the group consisting of camptothecin derivatives, camptothecin derivatives and α-amanitin.

49. A compound according to claim 46, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: The immune agonist is selected from TLR7 / 8 agonist or STING agonist.

50. A compound according to any one of claims 1 to 49, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: i is 2, 3, 4, 5, 6, 7 or 8.

51. A compound according to any one of claims 1 to 50, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: h is 1, 2, 3 or 4.

52. A compound according to any one of claims 1 to 51, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: o, p, q are 0, 1, 2 or 3 respectively.

53. A compound according to any one of claims 1 to 52, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: L a is one, two or more than three of a C1-C6 straight or branched alkylene group or a polyethylene glycol group, The polyethylene glycol group is -(CH2CH2O) n -、-O(CH2CH2O) n -、-(CH2CH2O) n CH2-, -(CH2CH2O) n CH2CH2-、-CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n CH2-, n is an integer greater than or equal to 1; Q a is one, two, or more than three of -C(=O)-, -NR-, -NR(C=O)-, and -(C=O)NR-, wherein, R is selected from hydrogen, C1-C6 straight chain or branched chain alkyl.

54. A compound according to claim 53, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: L a is ethylene, straight-chain propylene, straight-chain butylene, straight-chain pentylene, or -(CH2CH2O) n One, two or more of CH2CH2-, wherein n is 2, 3, 4, 5, 6, 7 or 8; Q a It is one, two, or more than three of -C(=O)-, -NR-, -NR(C=O)-, and -(C=O)NR-, wherein R is methyl and / or hydrogen.

55. A compound according to claim 53, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: L a is ethylene and / or -(CH2CH2O)4CH2CH2- or pentylene; Q a It is -C(=O)-, -N(CH3)-, -NH(C=O)-, -C(=O)NH- or -C(=O)-N(CH3)-.

56. A compound according to any one of claims 1 to 55, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: L b is selected from a C1-C6 straight chain or branched alkylene group, a C2-C6 straight chain or branched alkenylene group, or a C2-C6 straight chain or branched alkynylene group, an arylene group having 6 to 10 carbon atoms in one or more rings, or a polyethylene glycol group, The polyethylene glycol group is -(CH2CH2O) n -、-O(CH2CH2O) n -、-(CH2CH2O) n CH2-、-(CH2CH2O) n CH2CH2-、-CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n One or two or more than three of CH2-, n is an integer greater than or equal to 1, and the alkylene, alkenylene, and alkynylene groups are optionally substituted with substituents; Q b is selected from one, two or more of the group consisting of absence, -C(=O)NR-, -C(=O)O-, -OC(=O)NR-, -NRC(=O)-, -OC(=O)-, -NRC(=O)O-, -NRS(=O)2-, -NRC(=O)NR-, -C(=O)-, and -OC(=O)O-, and R is selected from hydrogen, an optionally substituted C1-C6 straight chain or branched alkyl group.

57. A compound according to claim 56, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: L b is a C1-C6 straight or branched chain alkylene group, or a C6-C 10 One or two or more than three kinds of arylene groups; Q b It is absent, one or two or more of -C(=O)NR-, -C(=O)-, -OC(=O)NR-, -NRC(=O)-, -NRC(=O)O-, or -NRC(=O)NR-, and R is selected from hydrogen, a C1-C6 straight chain or branched alkyl group.

58. A compound according to claim 56, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: L b It is one or two or more of the following: methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, sec-butylene, tert-butylene or benzylene; Q b is absent, -C(=O)-, -NRC(=O)-, -NRC(=O)NR- or -C(=O)NR-, or two or more thereof; and R is hydrogen or methyl.

59. A compound according to any one of claims 1 to 58, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: The substituent is selected from deuterium, tritium, a halogen atom, an amino group, a hydroxyl group, a cyano group, a nitro group, an optionally substituted alkyl group, an alkenyl group, an alkynyl group, an optionally substituted heterocyclic group, an acyl group, an optionally substituted amino group, an optionally substituted carbamoyl group, an optionally substituted thiocarbamoyl group, an optionally substituted sulfamoyl group, an optionally substituted hydroxyl group, an optionally substituted sulfanyl group (SH) and an optionally substituted silyl group.

60. A compound according to any one of claims 1 to 59, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: The compound is selected from the following structures:

61. A compound represented by formula (2), or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein: The structure of formula (2) is expressed as follows: Ab comprises an antibody or an antigen-binding fragment thereof; M' is a linking group; Sp is -L a -Q a -; A is selected from the group consisting of absent, hydrogen, and alkyl; T represents a linear or branched (C1-C8) trivalent alkyl group; Y, Y', and Y" are -L b -Q b -; L a , L b are respectively selected from absence, alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene, heteroarylene or polyethylene glycol group, wherein the alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene, heteroarylene, polyethylene glycol group is optionally substituted with a substituent; Q a , Q b are respectively selected from absent, -C(=O)NR-, -C(=O)O-, -OC(=O)NR-, -S(=O)2NR-, -NRC(=O)-, -OC(=O)-, -NRC(=O)O-, -NRS(=O)2-, -NRC(=O)NR-, -NHC-(=NH)NH-, -C(=O)-, -OC(=O)O-, -O-, -NR-, -S-, S(=O)2-, S(=O)-, or -Se-, wherein R is selected from hydrogen or optionally substituted alkyl; Z is selected from absent, hydrogen, optionally substituted alkyl or sulfonate; Z', Z" are selected from absent, hydrogen, optionally substituted alkyl, sulfonate or phosphate; At least one of Z, Z', and Z" is a sulfonic acid inner salt or a phosphate inner salt; X contains a self-destruct unit or does not exist; D includes drug molecules; i is an integer greater than or equal to 1; h is an integer from 1 to 6; o, p, q are integers from 0 to 5 respectively; j is an integer greater than or equal to 1.

62. A compound according to claim 61, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The formula (2) is formula (2-1): wherein Ab, M', Sp, A, T, Y, h, o, i, j, X, D, and Z are defined as in claim 61; At least one of the i Zs is a sulfonate.

63. A compound according to claim 62, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The formula (2-1) is formula (2-1-1): Among them, AA is the residue of amino acid, W is an integer greater than or equal to 1.

64. A compound according to claim 61, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The formula (2) is formula (2-2): Wherein, Ab, M', Sp, A, T, Y, Z, o, p, i, h, j, X, Y', and D are defined as in claim 61; and Z' is a sulfonic acid salt or a phosphate salt.

65. A compound according to claim 64, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The formula (2-2) is the formula (2-2-1): Wherein, AA is an amino acid residue, and W is an integer greater than or equal to 1.

66. A compound according to claim 61, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The formula (2) is formula (2-3): Wherein, Ab, M', Sp, A, Y, T, Z, o, i, h, j, q, X, Y", and D are defined as in claim 61; and Z" is a sulfonic acid salt or a phosphate salt.

67. A compound according to claim 66, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The formula (2-3) is formula (2-3-1): Wherein, AA is an amino acid residue, and W is an integer greater than or equal to 1.

68. A compound according to any one of claims 61-67, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: When A and Z do not exist, N, Y, and T form a ring, which is Preferably 69. A compound according to any one of claims 63, 65 or 67, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The amino acids are selected from one or more of alanine, arginine, asparagine, aspartic acid, γ-carboxyglutamate, citrulline, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, norleucine, norvaline, ornithine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine.

70. A compound according to any one of claims 63, 65 or 67, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The amino acids are selected from one or more of valine, glutamic acid, citrulline, lysine, alanine, phenylalanine, arginine, glutamine, asparagine and glycine.

71. A compound according to any one of claims 63, 65 or 67, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The amino acid is selected from one or more of valine, citrulline, alanine, phenylalanine, glutamine and glycine.

72. A compound according to any one of claims 61-71, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: M' is selected from one of the following structures: * Connect Ab.

73. A compound according to any one of claims 61-72, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: M' is selected from one of the following structures:

74. A compound according to any one of claims 61 to 73, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: M' is 75. A compound according to any one of claims 61-74, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: A is hydrogen, methyl, ethyl or propyl.

76. A compound according to any one of claims 61 to 75, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: A is hydrogen.

77. A compound according to any one of claims 61 to 76, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: The T is one of the following structures:

78. A compound according to any one of claims 61 to 77, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: The T is 79. A compound according to any one of claims 61 to 78, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: L a One, two or more selected from alkylene, alkenylene, alkynylene, saturated or unsaturated alicyclic group having 3-10 ring carbon atoms, saturated or unsaturated heteroalicyclic group having 3-10 ring carbon atoms and 1 to 4 heteroatoms, arylene group having 6 to 18 carbon atoms in one or more rings, or heteroarylene group having 6 to 18 carbon atoms in one or more rings and 1 to 4 heteroatoms, or polyethylene glycol group; the alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene group and heteroarylene group are optionally substituted by a substituent, and the heteroatom is selected from nitrogen, oxygen or sulfur.

80. A compound according to any one of claims 61-79, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: L a Selected from C1-C 10 Straight or branched alkylene, C2-C 12 Straight or branched alkenylene, C2-C 12 One, two or more of a straight or branched alkynylene group, a saturated or unsaturated alicyclic group having 3-6 ring carbon atoms, a saturated or unsaturated heteroalicyclic group having 3-6 ring carbon atoms and 1 to 4 heteroatoms, or an arylene group having 6 to 10 carbon atoms in one or more rings, a heteroarylene group having 6 to 10 carbon atoms in one or more rings and 1 to 4 heteroatoms, or a polyethylene glycol group, wherein the alkylene group, alkenylene group, alkynylene group, alicyclic group, heteroalicyclic group, arylene group and heteroarylene group are optionally substituted by a substituent, and the heteroatom is selected from nitrogen, oxygen or sulfur.

81. A compound according to any one of claims 61-80, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: L a One, two or more selected from C1-C6 straight chain or branched alkylene, C2-C6 straight chain or branched alkenylene, C2-C6 straight chain or branched alkynylene, or polyethylene glycol group, wherein the alkylene, alkenylene and alkynylene are optionally substituted by a substituent.

82. A compound according to any one of claims 61-81, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: L a is one, two or more than three of a C1-C6 straight or branched alkylene group or a polyethylene glycol group, The polyethylene glycol group is -(CH2CH2O) n -、-O(CH2CH2O) n -、-(CH2CH2O) n CH2-、-(CH2CH2O) n CH2CH2-、-CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n CH2-, n is an integer greater than or equal to 1.

83. A compound according to any one of claims 61-82, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: L a is ethylene, straight-chain propylene, straight-chain butylene, straight-chain pentylene, or -(CH2CH2O) n One, two or more of CH2CH2-, wherein n is 2, 3, 4, 5, 6, 7 or 8.

84. A compound according to any one of claims 61 to 83, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: L a It is ethylene and / or -(CH2CH2O)4CH2CH2- or pentylene.

85. A compound according to any one of claims 61-84, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: L b One or two or more selected from alkylene, alkenylene, alkynylene, saturated or unsaturated alicyclic group having 3-10 ring carbon atoms, saturated or unsaturated heteroalicyclic group having 3-10 ring carbon atoms and 1 to 4 heteroatoms, arylene group having 6 to 18 carbon atoms in one or more rings, or heteroarylene group having 6 to 18 carbon atoms in one or more rings and 1 to 4 heteroatoms, or polyethylene glycol, The alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene group, heteroarylene group may be substituted with a substituent, and the heteroatom may be selected from nitrogen, oxygen or sulfur.

86. A compound according to any one of claims 61-85, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: L b Selected from C1-C 10 Straight or branched alkylene, C2-C 12 Straight or branched alkenylene, C2-C 12 One or two or more of a linear or branched alkynylene group, a saturated or unsaturated alicyclic group having 3 to 6 ring carbon atoms, a saturated or unsaturated heteroalicyclic group having 3 to 6 ring carbon atoms and 1 to 4 heteroatoms, or an arylene group having 6 to 10 carbon atoms in one or more rings, a heteroarylene group having 6 to 10 carbon atoms in one or more rings and 1 to 4 heteroatoms, or polyethylene glycol, The alkylene, alkenylene, alkynylene, alicyclic group, heteroalicyclic group, arylene group, heteroarylene group may be substituted with a substituent, and the heteroatom may be selected from nitrogen, oxygen or sulfur.

87. A compound according to any one of claims 61-86, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: L b is selected from C1-C6 straight chain or branched chain alkylene, C2-C6 straight chain or branched chain alkenylene, or C2-C6 straight chain or branched chain alkynylene, or polyethylene glycol group, The polyethylene glycol group is -(CH2CH2O) n -、-O(CH2CH2O) n -、-(CH2CH2O) n CH2-、-(CH2CH2O) n CH2CH2-、-CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n One or two or more than three of CH2-, n is an integer greater than or equal to 1, The alkylene group, alkenylene group and alkynylene group may be substituted with a substituent.

88. A compound according to any one of claims 61-87, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: L b is a C1-C6 straight or branched chain alkylene group, or a C6-C 10 One or two or more than three of the arylene groups.

89. A compound according to any one of claims 61 to 88, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: L b It is one or two or more of methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, sec-butylene, tert-butylene, or benzylene.

90. A compound according to any one of claims 61-89, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: Q a One, two or more selected from -C(=O)NR-, -NR-, -C(=O)O-, -OC(=O)NR-, -NRC(=O)-, -OC(=O)-, -NRC(=O)O-, -NRC(=O)NR-, NHC-(=NH)NH-, -C(=O)-, -OC(=O)O-, wherein R is selected from C1-C6 straight chain or branched alkyl optionally substituted with hydrogen.

91. The compound of claim 90, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: Q a is one, two, or more than three of -C(=O)-, -NR-, -C(=O)NR-, -NRC(=O)NR-, -OC(=O)O-, -OC(=O)NR-, -NRC(=O)-, or -OC(=O)-, wherein, R is selected from hydrogen or an optionally substituted C1-C6 straight or branched chain alkyl group.

92. The compound according to claim 90, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein, Q a is one or two or more than three of -C(=O)-, -NR-, -NRC(=O)- and -(C=O)NR-, wherein, R is selected from hydrogen or an optionally substituted C1-C6 straight or branched chain alkyl group.

93. A compound according to claim 90, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: Q a It is -C(=O)-, -N(CH3)-, -NHC(=O)-, -C(=O)-NH- or -C(=O)-N(CH3)-.

94. A compound according to any one of claims 61-93, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: Q b is selected from one, two or more of the group consisting of absent, -C(=O)NR-, -C(=O)O-, -OC(=O)NR-, -NRC(=O)-, -OC(=O)-, -NRC(=O)O-, -NRC(=O)NR-, NHC-(=NH)NH-, -C(=O)-, and -OC(=O)O-, wherein R is selected from hydrogen or an optionally substituted C1-C6 straight or branched alkyl group.

95. A compound according to claim 94, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: Q b is absent, -C(=O)-, -C(=O)NR-, -OC(=O)NR-, -NRC(=O)-, -NRC(=O)O-, or -NRC(=O)NR-, and R is selected from hydrogen or an optionally substituted C1-C6 straight or branched alkyl group.

96. A compound according to claim 94, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: Q b is absent, -C(=O)-, -NRC(=O)-, -NRC(=O)NR- or -C(=O)NR-, or two or more thereof; and R is hydrogen or methyl.

97. A compound according to any one of claims 61-96, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: X is selected from: The right wavy line in X connects D; R4 is selected from hydrogen, C1-C6 straight or branched alkyl, -(CH2CH2O) m R2, -CH2CH2NR2R3, -CH2CH2SO2R2-, R2 and R3 are independently selected from hydrogen, C1-C6 straight chain or branched alkyl, and m is an integer greater than 1.

98. A compound according to claim 97, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: X is R4 is hydrogen, methyl, -CH2CH2OCH2CH2OH, -CH2CH2NMe2, -CH2CH2SO2Me.

99. The compound of claim 97, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein: X is R4 is hydrogen or methyl.

100. A compound according to any one of claims 61-99, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: The sulfonic acid salt is A wavy line connects Y, Y' or Y"; Wherein c, d, e, f, g are all integers of 1, 2, 3, 4, 5 or 6; R6, R7, R 11 , R 12 , R 13 , R 14 are independently selected from absence, hydrogen, and C1-C6 straight or branched chain alkyl.

101. A compound according to claim 100, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The sulfonic acid salt is in, R6, R7, R 11 , R 12 , R 13 and R 14 are respectively selected from absence, methyl or ethyl, c, d, e, f, and g are 1, 2, or 3 respectively.

102. A compound according to claim 100, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: The sulfonic acid inner salt is one of the following structures:

103. A compound according to any one of claims 61-102, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: The phosphate inner salt is in, a and b are both integers of 1, 2, 3, 4, 5 or 6, R8, R9 and R 10 are selected from hydrogen, C1-C6 straight chain or branched alkyl.

104. A compound according to claim 103, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The phosphate inner salt is in, R8, R9, R 10 They are methyl or ethyl respectively; a is 1 or 2 or 3; b is 1 or 2 or 3.

105. A compound according to claim 103, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The phosphate inner salt is in, R8, R9, R 10 All are methyl; a is 2; b is 1.

106. A compound according to any one of claims 61-105, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The D comprises a cytotoxin or an immune agonist.

107. A compound according to claim 106, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The cytotoxin is selected from one or two of a microtubule protein inhibitor, a DNA synthesis inhibitor, a topoisomerase inhibitor or an RNA polymerase 2 inhibitor.

108. A compound according to claim 106, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The cytotoxin is selected from auristatins, maytansine alkaloids, calicheamicin, duocarmycin, pyrrolobenzodiazepines, One or more of the group consisting of camptothecin derivatives, camptothecin derivatives and α-amanitin.

109. The compound according to claim 106, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein: The immune agonist is selected from TLR7 / 8 agonist or STING agonist.

110. A compound according to any one of claims 61 to 109, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: i is 2, 3, 4, 5, 6, 7 or 8.

111. A compound according to any one of claims 61 to 110, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: h is 1, 2, 3 or 4.

112. A compound according to any one of claims 61-111, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: o, p, q are 0, 1, 2 or 3 respectively.

113. A compound according to any one of claims 61-112, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: L a is one, two or more than three of a C1-C6 straight or branched alkylene group or a polyethylene glycol group, The polyethylene glycol group is -(CH2CH2O) n -、-O(CH2CH2O) n -、-(CH2CH2O) n CH2-、-(CH2CH2O) n CH2CH2-、-CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n CH2-, n is an integer greater than or equal to 1; Q a is one, two, or more than three of -C(=O)-, -NR(C=O)-, and -(C=O)NR-, wherein, R is selected from hydrogen, C1-C6 straight chain or branched chain alkyl.

114. A compound according to any one of claims 61 to 113, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: L a is ethylene, straight-chain propylene, straight-chain butylene, straight-chain pentylene, or -(CH2CH2O) n One, two or more of CH2CH2-, wherein n is 2, 3, 4, 5, 6, 7 or 8; Q a It is one, two, or more than three of -C(=O)-, -NR-, -NR(C=O)-, and -(C=O)NR-, wherein R is methyl and / or hydrogen.

115. A compound according to claim 114, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: L a is ethylene and / or -(CH2CH2O)4CH2CH2- or pentylene; Q a It is -C(=O)-, -N(CH3)-, -NH(C=O)-, -C(=O)NH- or -C(=O)-N(CH3)-.

116. A compound according to any one of claims 61 to 115, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: L b is selected from a C1-C6 straight chain or branched alkylene group, a C2-C6 straight chain or branched alkenylene group, or a C2-C6 straight chain or branched alkynylene group, an arylene group having 6 to 10 carbon atoms in one or more rings, or a polyethylene glycol group, The polyethylene glycol group is -(CH2CH2O) n -、-O(CH2CH2O) n -、-(CH2CH2O) n CH2-、-(CH2CH2O) n CH2CH2-、-CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n One or two or more than three of CH2-, n is an integer greater than or equal to 1, and the alkylene, alkenylene, and alkynylene groups are optionally substituted with substituents; Q b is selected from one, two or more of the group consisting of absence, -C(=O)NR-, -C(=O)O-, -OC(=O)NR-, -NRC(=O)-, -OC(=O)-, -NRC(=O)O-, -NRS(=O)2-, -NRC(=O)NR-, -C(=O)-, and -OC(=O)O-, and R is selected from hydrogen, an optionally substituted C1-C6 straight chain or branched alkyl group.

117. The compound according to claim 116, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein: L b is a C1-C6 straight or branched chain alkylene group, or a C6-C 10 One or two or more than three kinds of arylene groups; Q b is absent, one or two or more of -C(=O)-, -C(=O)NR-, -OC(=O)NR-, -NRC(=O)-, -NRC(=O)O-, or -NRC(=O)NR-, and R is selected from hydrogen, an optionally substituted C1-C6 straight chain or branched alkyl.

118. The compound according to claim 116, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate, wherein: L b It is one or two or more of the following: methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, sec-butylene, tert-butylene or benzylene; Q b is absent, -C(=O)-, -NRC(=O)-, -NRC(=O)NR- or -C(=O)NR-, or two or more thereof; and R is hydrogen or methyl.

119. A compound according to any one of claims 61 to 118, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: The substituent is selected from deuterium, tritium, a halogen atom, an amino group, a hydroxyl group, a cyano group, a nitro group, an optionally substituted alkyl group, an alkenyl group, an alkynyl group, an optionally substituted heterocyclic group, an acyl group, an optionally substituted amino group, an optionally substituted carbamoyl group, an optionally substituted thiocarbamoyl group, an optionally substituted sulfamoyl group, an optionally substituted hydroxyl group, an optionally substituted sulfanyl group (SH) and an optionally substituted silyl group.

120. A compound according to any one of claims 61-119, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: j is an integer from 1 to 20.

121. A compound according to any one of claims 61-120, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The compound is selected from the following structures:

122. A compound according to any one of claims 61-121, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The antibody is selected from the group consisting of a murine antibody, a chimeric antibody, a humanized antibody and a fully human antibody.

123. A compound according to any one of claims 61-122, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The antibodies comprise monoclonal antibodies.

124. A compound according to any one of claims 61-123, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The antibodies comprise bispecific antibodies.

125. A compound according to any one of claims 61-124, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The antigen binding fragment is selected from the group consisting of Fab, Fab', Fv fragment, F(ab')2, F(ab)2, scFv, di-scFv and VHH.

126. A compound according to any one of claims 61-125, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The Ab is selected from the following group: anti-claudin 18.2 antibody, anti-human folate receptor antibody, anti-HER2 antibody, anti-HER3 antibody, anti-TROP2 antibody, anti-B7-H3 antibody, anti-GPC20 antibody, anti-CDH6 antibody, anti-EGFR antibody, anti-EGFRvⅢ antibody, anti-AXL antibody, anti-Nectin-4 antibody, anti-Tissue factor antibody, anti-TIM-1 antibody, anti-PSMA antibody, anti-EpCAM antibody, anti-MUC1 antibody, anti-STEAP1 antibody, anti-GPNMB antibody, anti-FGF2 antibody, anti-FOLR1 antibody, anti-c-MET antibody, anti-GFR antibody, anti-AGS-16, anti-Guanylyl cyclase C antibody, anti-Mesothelin antibody, anti-SLC44A4 antibody, anti-EphA2 antibody, anti-AGS-5 antibody, anti-GPC-3 antibody, anti-c-KIT antibody, anti-ROR1 antibody, anti-PD-L1 antibody, anti-CD27L antibody, anti-5T4 antibody, anti-Mucin 16 antibody, anti-NaPi2b antibody, anti-STEAP antibody, anti-SLITRK6 antibody, anti-ETBR antibody, anti-BCMA antibody, anti-CEACAM5 antibody, anti-SC-16 antibody, anti-SLC39A6 antibody, anti-Delta-like protein3 antibody, anti-Claudin18.2 antibody, anti-CD19 antibody, anti-CD20 antibody, anti-CD22 antibody, anti-CD30 antibody, anti-CD33 antibody, anti-CD37 antibody, anti-CD45 antibody, anti-CD56 antibody, anti-CD66e antibody, anti-CD70 antibody, anti-CD73 antibody, anti-CD74 antibody, anti-CD79b antibody, anti-CD138 antibody, anti-CD147 antibody, anti-CD166 antibody, anti-CD223 antibody, anti-MUC16 antibody, anti-MSLN antibody, anti-ENPP3 antibody, anti-SLTRK6 antibody, anti-FGFR antibody, anti-LIV-1 antibody, anti-Lewis Y antibody, anti-av-integrin antibody, anti-ASCT2 antibody, anti-C4.4a antibody, anti-CA-IX antibody, anti-CD324 antibody, anti-CD352 antibody, anti-CD44v6 antibody, anti-CD48a antibody, anti-CLL-1 antibody, anti-Cripto antibody, anti-CS1 antibody, anti-DPEP3 antibody, anti-Ephrin-A2 antibody, anti-Ephrin-A4 antibody, anti-ETBR antibody, anti-FGFR2 antibody, anti-FGFR3 antibody, anti-FLT3 antibody, anti-GD3 antibody, anti-GloboH antibody, anti-GPC3 antibody, anti-LAMP-1 antibody, anti-LRRC15 antibody, anti-Ly6E antibody, anti-MFI2 antibody, anti-NOTCH3 antibody, anti-p-cadherin antibody, anti-PRLR antibody and anti-RNF43 antibody, and antigen-binding fragments of the above antibodies.

127. A compound according to any one of claims 61-126, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The heavy chain HCDR1, HCDR2 and HCDR3 and the light chain LCDR1, LCDR2 and LCDR3 of Ab comprise the heavy chain HCDR1, HCDR2 and HCDR3 and the light chain LCDR1, LCDR2 and LCDR3 of antibody, respectively.

128. A compound according to any one of claims 61-127, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The heavy chain variable region VH and light chain variable region VL of the Ab respectively comprise the heavy chain variable region VH and light chain variable region VL of the antibody.

129. A compound according to any one of claims 61-128, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The heavy chain and light chain of the Ab comprise the heavy chain and light chain of an antibody, respectively.

130. A compound according to any one of claims 61-129, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein: The Ab is an anti-claudin 18.2 antibody or an antigen-binding fragment thereof.

131. A compound according to any one of claims 61-130, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: The Ab is an anti-human folate receptor antibody or an antigen-binding fragment thereof.

132. A pharmaceutical composition, wherein It includes the compound according to any one of claims 1 to 131, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate.

133. Use of the compound described in any one of claims 1-132, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, prodrug or solvate in the preparation of a medicament for treating and / or preventing tumors.

134. The use according to claim 133, wherein The tumor is selected from tumors associated with the expression of the following group of targets: claudin 18.2, human folate receptor, HER2, HER3, TROP2, B7-H3, GPC20, CDH6, EGFR, EGFRvⅢ, AXL, Nectin-4, Tissue factor, TIM-1, PSMA, EpCAM, MUC1, STEAP1, GPNMB, FGF2, FOLR1, c-MET, GFR, AGS-16, Guanylyl cyclase C, Mesothelin, SLC44A4, EphA2, AGS-5, GPC-3, c-KIT, ROR1, PD-L1, CD27L, 5T4, Mucin 16, NaPi2b, STEAP, SLITRK6, ETBR, BCMA, CEACAM5, SC-16, SLC39A6, Delta-like protein3, Claudin18.2, CD19, CD20, CD22, CD30, CD33, CD37, CD45, CD56, CD66e, CD70, CD73, CD74, CD79b, CD138, CD147, CD166, CD223, MUC16, MSLN, ENPP3, SLTRK6, FGFR, LIV-1, Lewis Y, av-integrin, ASCT2, C4.4a, CA-IX, CD324, CD352, CD44v6, CD48a, CLL-1, Cripto, CS1, DPEP3, Ephrin-A2, Ephrin-A4, ETBR, FGFR2, FGFR3, FLT3, GD3, Globo H, GPC3, LAMP-1, LRRC15, Ly6E, MFI2, NOTCH3, p-cadherin, PRLR, and RNF43.

135. The use according to claim 133, wherein Tumors associated with the target expression include tumors with high expression of the target and / or target-positive tumors.

136. The use according to claim 133, wherein The tumor is selected from solid tumors, hematological tumors, and metastatic, refractory or recurrent lesions of cancer.

137. The use according to claim 133, wherein The tumor is selected from the group consisting of esophageal cancer, gastrointestinal cancer, pancreatic cancer, thyroid cancer, colorectal cancer, renal cancer, lung cancer (e.g., non-small cell lung cancer), liver cancer, gastric cancer, gastric adenocarcinoma, gastroesophageal junction (GEJ) adenocarcinoma, head and neck cancer, bladder cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, germ cell cancer, bone cancer, skin cancer, thymic cancer, bile duct cancer, gallbladder cancer, melanoma, mesothelioma, lymphoma, myeloma (e.g., multiple myeloma), sarcoma, glioblastoma, and leukemia.