Linker-drug molecules and antibody-drug conjugates, methods for their preparation and use

Abstract

Disclosed are compounds represented by formula (1), or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof. The structure of formula (1) is shown below. By introducing strongly hydrophilic sulfonic acid inner salts and / or phosphate inner salt side chains at the polypeptide (VC, VA, AAA, etc.)-PAB (self-destructing group) position of the linker, the compounds significantly improve the druggability (including hydrophilicity and stability) of the antibody-drug conjugates corresponding to the compounds, reduce drug clearance in the body, and enhance tumor therapeutic effects. JPEG2026523005000202.jpg3882

Description

[Technical Field]

[0001] This application relates to the field of biopharmaceuticals, and more specifically to linker-drug molecules and antibody-drug conjugates, methods for preparing them, and their use.

[0002] Cross-reference of related applications This application claims the interests of PCT / CN2023 / 106385, PCT International Application No. 7, July 2023, the contents of which are incorporated herein by reference in whole.

[0003] The sequence listing documents submitted at the same time All contents of the XML document below are incorporated herein by reference as a whole and are a computer-readable format (CRF) sequence listing (name: TFH01015PCT-sequence-listing.xml, date: 2024.7.2, size: 6KB). [Background technology]

[0004] By conjugating biologically active drug compounds to tumor-targeting antibodies, proteins, or polypeptides, the window of drug compounds for cancer treatment can be broadened. This is achieved primarily through two aspects. Firstly, because the active drug is not smoothly separated, the drug conjugate does not produce toxic side effects before reaching the cancer target lesion. Secondly, the active drug increases the concentration of the drug compound in the cancer target lesion via the conjugated targeted antibody, protein, or polypeptide, and a specially designed linker is stimulated and cleaved by the specific environment of the cancer target lesion, releasing the active drug compound.

[0005] The linker's specially designed cathepsin-sensitive amino acid peptide segment can be efficiently cleaved by cathepsin within the lysosomes of cancer cells. The drug compound is linked to the linker's amino acid peptide segment via a chemical self-destructing group. When cathepsin cleaves the peptide, the chemical self-destructing group releases the free drug compound through a "self-destructive effect," resulting in a tumor-killing effect. Among these, antibody or polypeptide drug conjugates with linkers in the form of polypeptides (VC, VA, AAA, etc.)-PAB (self-destructive group) are the most numerous in terms of clinically validated data and approved drugs. However, drug compounds, especially those with a "bystander-killing effect" crucial for tumor-killing, are highly lipophilic, and therefore antibody, protein, or polypeptide drug conjugates exhibit low druggability, particularly at high drug-antibody ratios (DAR), manifesting as decreased hydrophilicity, aggregate formation, and accelerated clearance in the body. To shield drug compounds, the druggability of antibody, protein, or polypeptide drug conjugates can be improved to some extent by adding hydrophilic groups 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), or sugar alcohols (Cancer Res (2018) 78 (13_Supplement): 755; US 9907854) to the linker. However, the use of PEG polymers to modify proteins has revealed many drawbacks in clinical practice, including a high proportion of patients with anti-PEG immunogenicity and related toxic side effects, and the impairment of protein function due to excessive modification. [Overview of the project] [Problems that the invention aims to solve]

[0006] Based on this, the present invention introduces a strongly hydrophilic sulfonic acid internal salt and / or phosphate internal salt side chain (hereinafter also referred to as the linker or internal salt linker of the present invention, i.e., the compound shown in formula (1) below) at the polypeptide (VC, VA, AAA, etc.)-PAB (self-destructing group) position of the linker. Since the sulfonic acid group or phosphate group has both a hydrogen bond donor and a hydrogen bond acceptor, it is a strongly hydrophilic group. Furthermore, since the sulfonic acid internal salt or phosphate internal salt has both a positive and negative charge, the hydrophilic properties can be further enhanced. In addition, the water-soluble group in the form of the internal salt may enhance tumor targeting of the antibody-drug conjugate and reduce nonspecific binding, thereby significantly improving the drugability (including hydrophilicity and stability) of the antibody-drug conjugate under test, reducing drug clearance in the body, and increasing the tumor therapeutic effect. [Means for solving the problem]

[0007] The technical solution of this application is as follows:

[0008] 1. The compound represented by formula (1), whose structure is shown below, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0009] [ka]

[0010] (M is a linking group, Sp is -L a -Q a -and, A is selected from none (non-existent), hydrogen, and alkyl. T represents a linear or branched (C1-C8) trivalent alkyl group. Y, Y', and Y'' are -L respectively. b -Q b -and, L a , Lb Each of these is selected from none, alkylene, alkenylene, alkylene, aliphatic cyclylene (alcylene, alcylene), aliphatic heterocyclylene, arylene, heteroarylene, or polyethylene glycol group, where the alkylene, alkenylene, alkylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, heteroarylene, and polyethylene glycol group are optionally substituted with substituents. Q a Q b Each of these is selected from none, -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-, where R is selected from hydrogen or optionally substituted alkyl. Z is selected from none, hydrogen, or optionally substituted alkyl or sulfonic acid internal salts. Z', Z'' are selected from none, hydrogen, optionally substituted alkyl, sulfonic acid end salts, or phosphate end salts. At least one of Z, Z', and Z'' is an end salt of a sulfonic acid or an end salt of a phosphate. X contains or does not contain a self-destructing unit. D contains drug molecules, i is an integer greater than or equal to 1, h is an integer between 1 and 6. o, p, and q are integers between 0 and 5.

[0011] 2. The compound described in item 1, wherein formula (1) is formula (1-1), or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0012] [ka]

[0013] (Here, the definitions of M, Sp, A, T, Y, h, o, i, X, D, and Z are as given in Section 1.) At least one of the i Zs is an intrasulfonic acid salt.

[0014] 3. The compound described in item 2, wherein formula (1-1) is formula (1-1-1), or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0015] [ka]

[0016] (Here, AA is an amino acid residue, and W is an integer greater than or equal to 1.)

[0017] 4. The compound described in item 1, wherein formula (1) is formula (1-2), or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0018] [ka]

[0019] (Here, the definitions of M, Sp, A, T, Y, Z, o, p, i, h, X, Y', and D are as in Section 1, and Z' is an endosodium sulfonic acid or endosodium phosphate.)

[0020] 5. The compound described in item 4, wherein formula (1) is formula (1-2-1), or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0021] [ka]

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

[0023] 6. The compound described in item 1, wherein formula (1) is formula (1-3), or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0024] [ka]

[0025] (Here, the definitions of M, Sp, A, Y, T, Z, o, i, h, q, X, Y'', and D are as in Section 1, and Z'' is an end salt of a sulfonic acid or an end salt of a phosphate.)

[0026] 7. The compound described in item 6, wherein formula (1) is formula (1-3-1), or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0027] [ka]

[0028] (Here, AA is an amino acid residue, and W is an integer greater than or equal to 1.)

[0029] 8. If both A and Z are absent, then N, Y, and T form a ring, i.e.

[0030] [ka]

[0031] Preferably

[0032] [ka]

[0033] The compounds described in any one of items 1 to 7, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0034] 9. The amino acids are one or more selected from alanine, arginine, asparagine, aspartic acid, γ-carboxyglutamic acid, citrulline, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, n-leucine, norvaline, ornithine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine, and are compounds described in any one of items 3, 5, or 7, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0035] 10. The amino acid is one or more selected from valine, glutamic acid, citrulline, lysine, alanine, phenylalanine, arginine, glutamine, asparagine, and glycine, and is a compound described in any one of items 3, 5, or 7, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0036] 11. The amino acid is one or more selected from valine, citrulline, alanine, phenylalanine, glutamine, and glycine, the compound according to any one of items 3, 5, or 7, or its tautomer, meso form, racemic form, enantiomer, diastereomer, a form of a mixture thereof, or its pharmaceutically acceptable salt, prodrug, or solvate.

[0037] 12. M is N3, SH, ONH2, NH2, CO2H, (alkyl)-CO-, HCO-, BrCH 2- , ICH2-,

[0038]

Chemical formula

[0039] The compound according to any one of items 1 to 11, or its tautomer, meso form, racemic form, enantiomer, diastereomer, a form of a mixture thereof, or its pharmaceutically acceptable salt, prodrug, or solvate, selected from

[0040] 13. M is

[0041]

Chemical formula

[0042] The compound according to any one of items 1 to 12, or its tautomer, meso form, racemic form, enantiomer, diastereomer, a form of a mixture thereof, or its pharmaceutically acceptable salt, prodrug, or solvate, and is

[0043] 14. M is

[0044]

Chemical formula

[0045] The compounds described in any one of items 1 to 13, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0046] 15.A is a compound described in any one of items 1 to 14, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein A is hydrogen, methyl, ethyl, or propyl.

[0047] 16.A is hydrogen, a compound described in any one of items 1 to 15, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0048] 17. The T is one of the following structures, a compound described in any one of the items 1 to 16, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0049] [ka]

[0050] 18. The above T is

[0051] [ka]

[0052] The compounds described in any one of items 1 to 17, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0053] 19.L a This refers to one, two, or three or more compounds selected from alkylene, alkenylene, alkynylene, saturated or unsaturated aliphatic cyclylene having 3 to 10 ring carbon atoms, saturated or unsaturated aliphatic heterocyclylene having 3 to 10 ring carbon atoms and 1 to 4 heteroatoms, arylene having 6 to 18 carbon atoms with one or more rings, heteroarylene having 6 to 18 carbon atoms and 1 to 4 heteroatoms with one or more rings, or polyethylene glycol groups, wherein the alkylene, alkenylene, alkynylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, and heteroarylene are optionally substituted by substituents, and the heteroatoms are selected from nitrogen, oxygen, or sulfur, as described in any one of claims 1 to 18, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0054] 20L a C1~C 10 Straight-chain or branched-chain alkylenes, C2-C 12 Straight-chain or branched-chain alkenylenes, C2-C 12 One, two, or three or more compounds selected from linear or branched alkylenes, saturated or unsaturated aliphatic cyclylenes having 3 to 6 ring carbon atoms, saturated or unsaturated aliphatic heterocyclylenes containing heteroatoms and having 3 to 6 ring carbon atoms and having 1 to 4 heteroatoms, arylenes having 6 to 10 carbon atoms and having one or more rings, heteroarylenes having 6 to 10 carbon atoms and having 1 to 4 heteroatoms, or polyethylene glycol groups, wherein the alkylenes, alkenylenes, alkylenes, aliphatic cyclylenes, aliphatic heterocyclylenes, arylenes, and heteroarylenes are optionally substituted by substituents, and the heteroatoms are selected from nitrogen, oxygen, or sulfur, as described in any one of claims 1 to 19, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0055] 21.L a The compound described in any one of claims 1 to 20, or in the form of a tautomer, mesomer, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt thereof, prodrug, or solvate thereof, wherein the alkylene, alkenylene, or alkylene is optionally substituted by substituents.

[0056] 22.L a This is one, two, or three or more C1-C6 linear or branched alkylenes, or polyethylene glycol groups, wherein the polyethylene glycol groups are -(CH2CH2O) n -, -O(CH2CH2O) n -,-(CH2CH2O) n CH2-, -(CH2CH2O) n CH2CH2-, -CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n A compound described in any one of items 1 to 21, wherein the compound is CH2- and n is an integer of 1 or more, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0057] 23.L a This refers to ethylene, linear propylene, linear butylene, linear pentylene, or -(CH2CH2O) n A compound described in any one of items 1 to 22, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein n is one, two or more of CH2CH2-, where n is 2, 3, 4, 5, 6, 7, or 8.

[0058] 24.L a The compound described in any one of the items 1 to 23, which is ethylene and / or -(CH2CH2O)4CH2CH2- or pentylene, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0059] 25L b This refers to one, two, or three or more compounds selected from alkylene, alkenylene, alkynylene, saturated or unsaturated aliphatic cyclylene having 3 to 10 ring carbon atoms, saturated or unsaturated aliphatic heterocyclylene having 3 to 10 ring carbon atoms and 1 to 4 heteroatoms, arylene having 6 to 18 carbon atoms with one or more rings, heteroarylene having 6 to 18 carbon atoms and 1 to 4 heteroatoms with one or more rings, or polyethylene glycol, wherein the alkylene, alkenylene, alkynylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, or heteroarylene is optionally substituted by substituents, and the heteroatoms are selected from nitrogen, oxygen, or sulfur, as described in any one of claims 1 to 24, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0060] 26.L b C1~C 10 Straight-chain or branched-chain alkylenes, C2-C 12 Straight-chain or branched-chain alkenylenes, C2-C 12One, two, or three or more compounds selected from linear or branched alkylylenes, saturated or unsaturated aliphatic cyclylenes having 3 to 6 ring carbon atoms, saturated or unsaturated aliphatic heterocyclylenes containing heteroatoms and having 3 to 6 ring carbon atoms and having 1 to 4 heteroatoms, arylenes having 6 to 10 carbon atoms and having one or more rings, heteroarylenes having 6 to 10 carbon atoms and having 1 to 4 heteroatoms, or polyethylene glycol, wherein the alkylenes, alkenylenes, alkylylenes, aliphatic cyclylenes, aliphatic heterocyclylenes, arylenes, and heteroarylenes are optionally substituted by substituents, and the heteroatoms are selected from nitrogen, oxygen, or sulfur, as described in any one of claims 1 to 25, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0061] 27.L b The group is selected from C1-C6 linear or branched alkylene, C2-C6 linear or branched alkenylene, C2-C6 linear 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 A compound according to any one of claims 1 to 26, wherein one, two or three or more CH2- groups are present, n is an integer of 1 or more, and the alkylene, alkenylene, or alkynylene is optionally substituted by substituents, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0062] 28.L bThis refers to C1-C6 straight-chain or branched-chain alkylenes, or C6-C6 10 Compounds described in any one of items 1 to 27, which are one, two or more allirenes, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0063] 29.L b This refers to one, two or more of the compounds described in any one of items 1 to 28, which are methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, sec-butylene, tert-butylene, or benzylene, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0064] 30.Q a is 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-, where R is selected from hydrogen, optionally substituted (may be substituted) C1-C6 linear or branched alkyl, one of the compounds described in any one of items 1-29, or in the form of tautomers, meso, racemics, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0065] 31.Q ais one, two or more of -C(=O)-, -NR-, -C(=O)NR-, -NRC(=O)NR-, -OC(=O)O-, -OC(=O)NR-, -NRC(=O)- or -OC(=O)-, where R is selected from hydrogen or optionally substituted C1-C6 linear or branched alkyl groups, the compound described in item 30, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0066] 32.Q a is one, two or more of -C(=O)-, -NR-, -NRC(=O)-, and -(C=O)NR-, where R is selected from hydrogen or optionally substituted C1-C6 linear or branched alkyl groups, the compound described in item 30, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0067] 33.Q a The compounds described in item 30, which are -C(=O)-, -N(CH3)-, -NHC(=O)-, -C(=O)-NH-, or -C(=O)-N(CH3)-, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0068] 34.Q bis one, two or more selected from none, -C(=O)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-, where R is selected from hydrogen, optionally substituted C1-C6 linear or branched alkyl, one of the compounds described in any one of items 1-33, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0069] 35.Q b is one, two or more of -C(=O)-, -C(=O)NR-, -OC(=O)NR-, -NRC(=O)-, -NRC(=O)O-, or -NRC(=O)NR-, where R is selected from hydrogen, optionally substituted C1-C6 linear or branched alkyl groups, the compounds described in item 34, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0070] 36.Q b The compound described in item 34, or its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof, wherein R is none, -C(=O)-, -NRC(=O)-, -NRC(=O)NR-, or -C(=O)NR-, and R is one, two or three or more of these, where R is hydrogen or methyl.

[0071] 37.X is a compound described in any one of sections 1 to 36, selected from the following: a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0072] [ka]

[0073] (The wavy line on the right in X is connected to D, R4 is hydrogen, C1-C6 straight or branched alkyl, -(CH2CH2O) m R2 is selected from -CH2CH2NR2R3 and -CH2CH2SO2R2, where R2 and R3 are selected from hydrogen and a C1-C6 straight-chain or branched-chain alkyl group, respectively, and m is an integer greater than 1.

[0074] 38.X is,

[0075] [ka]

[0076] And, R4 is hydrogen, methyl, -CH2CH2OCH2CH2OH, -CH2CH2NMe2, -CH2CH2SO2Me, or a compound as described in item 37, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0077] 39.X is,

[0078] [ka]

[0079] The compound described in item 37, or its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof, wherein R4 is hydrogen or methyl.

[0080] 40. The sulfonic acid salt is one of the following: a compound described in any one of items 1 to 39, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0081] [ka]

[0082] (The wavy line is connected to Y, Y', or Y'', Here, c, d, e, f, and g are all integers of 1, 2, 3, 4, 5, or 6. R6, R7, R 11 , R 12 , R 13 , R 14 (Each element is selected from none, hydrogen, or a C1-C6 linear or branched alkyl group.)

[0083] 41. The sulfonic acid salt is one of the compounds described in item 40, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0084] [ka]

[0085] (Here, R6, R7, R 11 , R 12 , R 13 and R 14 Each is selected from none, methyl, or ethyl. c, d, e, f, and g are 1, 2, or 3, respectively.

[0086] 42. The sulfonic acid salt is one of the following structures, a compound as described in item 40, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0087] [ka]

[0088] 43. The phosphate salt is one of the compounds described in any one of items 1 to 42, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0089] [ka]

[0090] (Here, a and b are all integers of 1, 2, 3, 4, 5, or 6, and R8, R9, and R 10 (Each element is selected from hydrogen, a C1-C6 linear chain, or a branched-chain alkyl group.)

[0091] 44. The phosphate salt is one of the compounds described in item 43, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0092] [ka]

[0093] (Here, R8, R9, R 10 (wherein 1 is methyl or ethyl, and 'a' is 1, 2, or 3, and 'b' is 1, 2, or 3.)

[0094] 45. The inner salt of phosphoric acid is the compound described in item 43, or its tautomer, meso form, racemic form, enantiomer, diastereomer, a mixture thereof, or its pharmaceutically acceptable salt, prodrug, or solvate.

[0095]

Chemical formula

[0096] (where R8, R9, R 10 are all methyl, a is 2, and b is 1.)

[0097] 46. D in the compound according to any one of items 1 to 45, or its tautomer, meso form, racemic form, enantiomer, diastereomer, a mixture thereof, or its pharmaceutically acceptable salt, prodrug, or solvate, contains a cytotoxin or an immune agonist.

[0098] 47. The cytotoxin in the compound according to item 46, or its tautomer, meso form, racemic form, enantiomer, diastereomer, a mixture thereof, or its pharmaceutically acceptable salt, prodrug, or solvate, is one or two selected from tubulin inhibitors, DNA synthesis inhibitors, topoisomerase inhibitors, or RNA polymerase 2 inhibitors.

[0099] 48. The cytotoxin in the compound according to item 46, or its tautomer, meso form, racemic form, enantiomer, diastereomer, a mixture thereof, or its pharmaceutically acceptable salt, prodrug, or solvate, is one or more selected from auristatins, maytansinoid DMs, calicheamicin, duocarmycin, pyrrolobenzodiazepines, camptothecin derivatives, and α - amanitin.

[0100] 49. The immunoagonist is a compound selected from a TLR7 / 8 agonist or a STING agonist, as described in item 46, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0101] 50.i is a compound described in any one of the items 1 to 49, which is 2, 3, 4, 5, 6, 7, or 8, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0102] 51.h is a compound described in any one of the items 1 to 50, where h is 1, 2, 3 or 4, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0103] 52. A compound described in any one of items 1 to 51, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, where o, p, and q are 0, 1, 2, or 3, respectively.

[0104] 53.L a This consists of one, two, or three or more C1-C6 linear or branched alkylene groups, or polyethylene glycol groups. The aforementioned polyethylene glycol group is -(CH2CH2O) n -, -O(CH2CH2O) n -,-(CH2CH2O) n CH2-, -(CH2CH2O) n CH2CH2-, -CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n CH2-, where n is an integer greater than or equal to 1. Q ais one, two, or three or more of -C(=O)-, -NR-, -NR(C=O)-, and -(C=O)NR-, where, R is a compound described in any one of items 1 to 52, selected from hydrogen, C1-C6 linear or branched alkyl groups, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0105] 54.L a This refers to ethylene, linear propylene, linear butylene, linear pentylene, or -(CH2CH2O) n It consists of one, two, or three or more of the CH2CH2- groups, where n is 2, 3, 4, 5, 6, 7, or 8, and Q a The compound described in item 53, or its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof, wherein R is one, two, or three or more of -C(=O)-, -NR-, -NR(C=O)-, and -(C=O)NR-, where R is methyl and / or hydrogen.

[0106] 55.L a These are ethylene and / or -(CH2CH2O)4CH2CH2- or pentylene, Q a The compounds described in item 53, which are -C(=O)-, -N(CH3)-, -NH(C=O)-, -C(=O)NH-, or -C(=O)-N(CH3)-, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0107] 56.L bThis is selected from C1-C6 linear or branched alkylenes, C2-C6 linear or branched alkenylenes, C2-C6 linear or branched alkynylenes, C6-C6 linear or branched alkynylenes, arylenes having one or more rings and 6-10 carbon atoms, or polyethylene glycol groups. The aforementioned polyethylene glycol group is -(CH2CH2O) n -, -O(CH2CH2O) n -,-(CH2CH2O) n CH2-, -(CH2CH2O) n CH2CH2-, -CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n It consists of one, two, or three or more CH2- groups, where n is an integer of 1 or more, and the alkylene, alkenylene, and alkynylene are optionally substituted by substituents. Q b is one, two or more selected from none, -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-, where R is selected from hydrogen, optionally substituted C1-C6 linear or branched alkyl, a compound described in any one of items 1-55, or its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0108] 57.L b This refers to C1-C6 straight-chain or branched-chain alkylenes, or C6-C6 10 It is one, two, or three or more of the allerenes. Q bis one, two or more of -C(=O)NR-, -C(=O)-, -OC(=O)NR-, -NRC(=O)-, -NRC(=O)O-, or -NRC(=O)NR-, where R is selected from hydrogen, C1-C6 linear or branched alkyl, one of the compounds described in item 56, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0109] 58.L b This is one, two, or three or more of methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, sec-butylene, tert-butylene, or bendiylene. Q b The compound described in item 56, or its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof, wherein R is none, -C(=O)-, -NRC(=O)-, -NRC(=O)NR-, or -C(=O)NR-, and R is one, two or three or more of these, where R is hydrogen or methyl.

[0110] 59. The substituents are selected from deuterium, tritium, halogen atoms, amino, hydroxy, cyano, nitro, optionally substituted alkyl, alkenyl, alkynyl groups, optionally substituted cyclyl groups, acyl, optionally substituted amino, optionally substituted carbamoyl, optionally substituted thiocarbamoyl, optionally substituted sulfamoyl, optionally substituted hydroxy, optionally substituted thioalkyl (SH), and optionally substituted silyl, and are compounds described in any one of items 1 to 58, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0111] 60. The compound is the compound according to any one of items 1 to 59 selected from the following structures, or its tautomer, meso form, racemic form, enantiomer, diastereomer, in the form of a mixture thereof, or its pharmaceutically acceptable salt, prodrug, or solvate.

[0112]

Chem.

[0113]

Chem.

[0114]

Chem.

[0115] 61. The compound represented by formula (2), or its tautomer, meso form, racemic form, enantiomer, diastereomer, in the form of a mixture thereof, or its pharmaceutically acceptable salt, prodrug, or solvate, wherein the structure of formula (2) is shown below.

[0116]

Chem.

[0117] (Ab includes an antibody or an antigen-binding fragment thereof, M ’ is a linking group, Sp is -L a -Q a -, A is selected from none, hydrogen, and alkyl, T represents a linear or branched (C1-C8) trivalent alkyl group, Y, Y’, Y” are each -L b -Q b -, L a L bEach of these is selected from none, alkylene, alkenylene, alkylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, heteroarylene, or polyethylene glycol group, where the alkylene, alkenylene, alkylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, heteroarylene, and polyethylene glycol group are optionally substituted with substituents. Q a Q b Each of these is selected from none, -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-, where R is selected from hydrogen or optionally substituted alkyl. Z is selected from none, hydrogen, or optionally substituted alkyl or sulfonic acid internal salts. Z', Z'' are selected from none, hydrogen, optionally substituted alkyl, sulfonic acid end salts, or phosphate end salts. At least one of Z, Z', and Z'' is an end salt of a sulfonic acid or an end salt of a phosphate. X includes self-destructing units or none. D contains drug molecules, i is an integer greater than or equal to 1, h is an integer between 1 and 6. o, p, and q are integers from 0 to 5, j is an integer greater than or equal to 1.

[0118] 62. The compound described in item 61, wherein formula (2) is formula (2-1), or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0119] [ka]

[0120] (Here, Ab, M ’ The definitions of Sp, A, T, Y, h, o, i, j, X, D, and Z are as given in section 61. At least one of the i Zs is an intrasulfonic acid salt.

[0121] 63. The compound described in item 62, wherein formula (2-1) is formula (2-1-1), or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0122] [ka]

[0123] (Here, AA is an amino acid residue, W is an integer greater than or equal to 1.

[0124] 64. The compound described in item 61, wherein formula (2) is formula (2-2), or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0125] [ka]

[0126] (Here, Ab, M ’ The definitions of Sp, A, T, Y, Z, o, p, i, h, j, X, Y', and D are as given in Section 61, where Z' is an endosodium sulfonic acid or endosodium phosphate.

[0127] The compound according to item 64, or a tautomer, meso form, racemic form, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein the formula (2-2) is formula (2-2-1).

[0128] [Chemical formula]

[0129] (where AA is a residue of an amino acid and W is an integer of 1 or more.)

[0130] The compound according to item 61, or a tautomer, meso form, racemic form, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein the formula (2) is formula (2-3).

[0131] [Chemical formula]

[0132] (where the definitions of Ab, M ’ , Sp, A, Y, T, Z, o, i, h, j, q, X, Y", D are as defined in item 61, and Z" is an inner salt of sulfonic acid or an inner salt of phosphoric acid.)

[0133] The compound according to item 66, or a tautomer, meso form, racemic form, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein the formula (2-3) is formula (2-3-1).

[0134] [Chemical formula]

[0135] (where AA is a residue of an amino acid and W is an integer of 1 or more.)

[0136] 68. If both A and Z are absent, N, Y, and T form a ring, i.e.,

[0137] [ka]

[0138] Preferably

[0139] [ka]

[0140] The compounds described in any one of paragraphs 61 to 67, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0141] 69. The amino acids are one or more selected from alanine, arginine, asparagine, aspartic acid, γ-carboxyglutamic acid, citrulline, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, n-leucine, norvaline, ornithine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine, and are compounds described in any one of items 63, 65, or 67, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0142] 70. The amino acid is one or more selected from valine, glutamic acid, citrulline, lysine, alanine, phenylalanine, arginine, glutamine, asparagine, and glycine, the compound described in any one of claims 63, 65, or 67, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0143] 71. The amino acid is one or more selected from valine, citrulline, alanine, phenylalanine, glutamine, and glycine, the compound described in any one of claims 63, 65, or 67, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0144] 72.M ’ This refers to a compound described in any one of the following sections 61 to 71, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, selected from the structures described below.

[0145] [ka]

[0146] (* is concatenated to Ab.)

[0147] 73.M ’ This refers to a compound described in any one of the following sections 61 to 72, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, selected from the structures described below.

[0148] [ka]

[0149] 74.M ’ teeth,

[0150] [ka]

[0151] The compounds described in any one of paragraphs 61 to 73, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0152] 75.A is a compound described in any one of sections 61 to 74, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein A is hydrogen, methyl, ethyl, or propyl.

[0153] 76.A is hydrogen, a compound as described in any one of subsections 61 to 75, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0154] 77. The T is one of the structures described below, a compound described in any one of the items 61 to 76, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0155] [ka]

[0156] 78. The above T is,

[0157] [ka]

[0158] The compounds described in any one of paragraphs 61 to 77, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0159] 79.L a The compound is one, two, or three or more selected from alkylene, alkenylene, alkynylene, saturated or unsaturated aliphatic cyclylene having 3 to 10 ring carbon atoms, saturated or unsaturated aliphatic heterocyclylene having 3 to 10 ring carbon atoms and 1 to 4 heteroatoms, arylene having 6 to 18 carbon atoms with one or more rings, heteroarylene having 6 to 18 carbon atoms and 1 to 4 heteroatoms, or polyethylene glycol groups, wherein the alkylene, alkenylene, alkynylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, or heteroarylene is optionally substituted by substituents, and the heteroatoms are selected from nitrogen, oxygen, or sulfur, as described in any one of claims 61 to 78, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0160] 80L a C1~C 10 Straight-chain or branched-chain alkylenes, C2-C 12 Straight-chain or branched-chain alkenylenes, C2-C 12One, two, or three or more compounds selected from linear or branched alkylenes, saturated or unsaturated aliphatic cyclylenes having 3 to 6 ring carbon atoms, saturated or unsaturated aliphatic heterocyclylenes containing heteroatoms and having 3 to 6 ring carbon atoms and having 1 to 4 heteroatoms, arylenes having 6 to 10 carbon atoms and having one or more rings, heteroarylenes having 6 to 10 carbon atoms and having 1 to 4 heteroatoms, or polyethylene glycol groups, wherein the alkylenes, alkenylenes, alkylenes, aliphatic cyclylenes, aliphatic heterocyclylenes, arylenes, and heteroarylenes are optionally substituted by substituents, and the heteroatoms are selected from nitrogen, oxygen, or sulfur, as described in any one of claims 61 to 79, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0161] 81.L a The compound described in any one of claims 61 to 80, or in the form of a tautomer, mesomer, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt thereof, prodrug, or solvate thereof, wherein the alkylene, alkenylene, or alkylene is optionally substituted by substituents, and is a compound described in any one of claims 61 to 80, or a tautomer, mesomer, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt thereof.

[0162] 82.L a This consists of one, two, or three or more C1-C6 linear or branched alkylene groups, or polyethylene glycol groups. The aforementioned polyethylene glycol group is -(CH2CH2O) n -, -O(CH2CH2O) n -,-(CH2CH2O) n CH2-, -(CH2CH2O) n CH2CH2-, -CH2O(CH2CH2O) n-, or -CH2O(CH2CH2O) n A compound described in any one of items 61 to 81, which is CH2- and n is an integer of 1 or more, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0163] 83.L a This refers to ethylene, linear propylene, linear butylene, linear pentylene, or -(CH2CH2O) n A compound described in any one of items 61 to 82, comprising one, two or more CH2CH2- groups, where n is 2, 3, 4, 5, 6, 7, or 8, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0164] 84.L a The compound described in any one of the items 61 to 83, which is ethylene and / or -(CH2CH2O)4CH2CH2- or pentylene, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0165] 85.L b This is one, two, or three or more selected from alkylene, alkenylene, alkynylene, saturated or unsaturated aliphatic cyclylene with 3 to 10 ring carbon atoms, saturated or unsaturated aliphatic heterocyclylene with 3 to 10 ring carbon atoms and 1 to 4 heteroatoms, arylene with 6 to 18 carbon atoms having one or more rings, heteroarylene with 6 to 18 carbon atoms having one or more rings and 1 to 4 heteroatoms, or polyethylene glycol. The alkylene, alkenylene, alkynylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, heteroarylene is optionally substituted by substituents, wherein the heteroatom is selected from nitrogen, oxygen, or sulfur, and the compounds are as described in any one of claims 61 to 84, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0166] 86.L b C1~C 10 Straight-chain or branched-chain alkylenes, C2-C 12 Straight-chain or branched-chain alkenylenes, C2-C 12 One, two, or three or more selected from linear or branched alkynylenes, saturated or unsaturated aliphatic cyclylenes with 3 to 6 ring carbon atoms, saturated or unsaturated aliphatic heterocyclylenes containing heteroatoms and having 3 to 6 ring carbon atoms and 1 to 4 heteroatoms, arylenes with 6 to 10 carbon atoms having one or more rings, heteroarylenes with 6 to 10 carbon atoms and 1 to 4 heteroatoms having one or more rings, or polyethylene glycol. The alkylene, alkenylene, alkynylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, heteroarylene is optionally substituted by substituents, wherein the heteroatom is selected from nitrogen, oxygen, or sulfur, and the compounds are as described in any one of claims 61 to 85, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0167] 87.L b This is selected from C1-C6 linear or branched alkylene, C2-C6 linear or branched alkenylene, C2-C6 linear or branched alkylylene, or polyethylene glycol group. The aforementioned polyethylene glycol group is -(CH2CH2O) n -, -O(CH2CH2O) n-,-(CH2CH2O) n CH2-, -(CH2CH2O) n CH2CH2-, -CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n It consists of one, two, or three or more of the CH2- elements, where n is an integer greater than or equal to 1. The alkylene, alkenylene, and alkynylene are optionally substituted with substituents in the form of compounds according to any one of claims 61 to 86, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0168] 88.L b This refers to C1-C6 straight-chain or branched-chain alkylenes, or C6-C6 10 Compounds described in any one of the items 61 to 87, which are one, two or more allirenes, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0169] 89.L b This refers to one, two or more of methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, sec-butylene, tert-butylene, or benzylene, as described in any one of the compounds in any one of the items 61 to 88, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0170] 90.Q ais 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-, where R is selected from C1-C6 linear or branched alkyl groups with optionally substituted hydrogen, and is a compound described in any one of items 61-89, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0171] 91.Q a is one, two, or three or more of -C(=O)-, -NR-, -C(=O)NR-, -NRC(=O)NR-, -OC(=O)O-, -OC(=O)NR-, -NRC(=O)-, or -OC(=O)-, where, R is selected from C1-C6 linear or branched alkyl groups that are hydrogen or optionally substituted, and is one of the compounds described in item 90, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0172] 92.Q a is one, two, or three or more of -C(=O)-, -NR-, -NRC(=O)-, and -(C=O)NR-, where, R is selected from C1-C6 linear or branched alkyl groups that are hydrogen or optionally substituted, and is one of the compounds described in item 90, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0173] 93.Q aThe compounds described in item 90, which are -C(=O)-, -N(CH3)-, -NHC(=O)-, -C(=O)-NH-, or -C(=O)-N(CH3)-, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0174] 94.Q b is one, two or more selected from none, -C(=O)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-, where R is selected from hydrogen or optionally substituted C1-C6 linear or branched alkyl groups, one of the compounds described in any one of items 61-93, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0175] 95.Q b The compound described in item 94, or its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof, wherein R is none, -C(=O)-, -C(=O)NR-, -OC(=O)NR-, -NRC(=O)-, -NRC(=O)O-, or -NRC(=O)NR-, where R is selected from hydrogen or optionally substituted C1-C6 linear or branched alkyl groups, or its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or its pharmaceutically acceptable salts, prodrugs, or solvates.

[0176] 96.Q b The compound described in item 94, or its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof, wherein R is none, -C(=O)-, -NRC(=O)-, -NRC(=O)NR-, or -C(=O)NR-, and R is one, two or more of these, where R is hydrogen or methyl.

[0177] 97.X is a compound described in any one of sections 61 to 96, selected from the following: a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0178] [ka]

[0179] (The wavy line on the right in X is connected to D, R4 is hydrogen, C1-C6 straight or branched alkyl, -(CH2CH2O) m Selected from R2, -CH2CH2NR2R3, -CH2CH2SO2R2-, R2 and R3 are selected from hydrogen and a C1-C6 straight-chain or branched-chain alkyl group, respectively, and m is an integer greater than 1.

[0180] 98.X is,

[0181] [ka]

[0182] And, R4 is hydrogen, methyl, -CH2CH2OCH2CH2OH, -CH2CH2NMe2, -CH2CH2SO2Me, or a compound as described in item 97, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0183] 99.X is,

[0184] [ka]

[0185] The compound described in item 97, or its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof, wherein R4 is hydrogen or methyl.

[0186] 100. The sulfonic acid salt is one of the compounds described in any one of the items 61 to 99, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0187] [ka]

[0188] (The wavy line is connected to Y, Y', or Y'', Here, c, d, e, f, and g are all integers of 1, 2, 3, 4, 5, or 6. R6, R7, R 11 , R 12 , R 13 , R 14 (Each element is selected from none, hydrogen, or a C1-C6 linear or branched alkyl group.)

[0189] 101. The sulfonic acid salt is one of the compounds described in item 100, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0190] [ka]

[0191] (Here, R6, R7, R 11 , R 12 , R 13 and R 14 Each is selected from none, methyl, or ethyl. c, d, e, f, and g are 1, 2, or 3, respectively.

[0192] 102. The sulfonic acid internal salt is one of the following structures, the compound described in item 100, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0193] [ka]

[0194] 103. The phosphate salt is one of the compounds described in any one of items 61 to 102, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0195] [ka]

[0196] (Here, a and b are all integers of 1, 2, 3, 4, 5, or 6, and R8, R9, and R 10 (Each element is selected from hydrogen, a C1-C6 linear chain, or a branched-chain alkyl group.)

[0197] 104. The phosphate salt is one of the compounds described in item 103, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0198] [ka]

[0199] (Here, R8, R9, R 10 (wherein 1 is methyl or ethyl, and 'a' is 1, 2, or 3, and 'b' is 1, 2, or 3.)

[0200] 105. The phosphate salt is one of the compounds described in item 103, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0201] [ka]

[0202] (Here, R8, R9, R 10 (All are methyl, a is 2, and b is 1.)

[0203] 106. The D is a compound described in any one of the items 61 to 105, including a cytotoxin or immunoagonist, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0204] 107. The cytotoxin is one or two selected from tubulin inhibitors, DNA synthesis inhibitors, topoisomerase inhibitors, or RNA polymerase 2 inhibitors, the compound described in item 106, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0205] 108. The cytotoxin is one or more compounds selected from auristatins, meitansinoid DMs, calicheamicin, duocarmycin, pyrrolobenzodiazepines, camptothecin derivatives, and α-amanitin, as described in item 106, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0206] 109. The immunoagonist is a compound selected from a TLR7 / 8 agonist or a STING agonist, as described in subsection 106, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0207] 110.i is a compound described in any one of the sections 61 to 109, which is 2, 3, 4, 5, 6, 7, or 8, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0208] 111.h is 1, 2, 3 or 4, a compound as described in any one of the items 61 to 110, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0209] 112. A compound described in any one of the items 61 to 111, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, where o, p, and q are 0, 1, 2, or 3, respectively.

[0210] 113.L aThis consists of one, two, or three or more C1-C6 linear or branched alkylene groups, or polyethylene glycol groups. The aforementioned polyethylene glycol group is -(CH2CH2O) n -, -O(CH2CH2O) n -,-(CH2CH2O) n CH2-, -(CH2CH2O) n CH2CH2-, -CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n CH2-, where n is an integer greater than or equal to 1. Q a is one, two, or three or more of -C(=O)-, -NR(C=O)-, and -(C=O)NR-, where, R is a compound described in any one of items 61 to 112, selected from hydrogen, C1-C6 linear or branched alkyl groups, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0211] 114.L a This refers to ethylene, linear propylene, linear butylene, linear pentylene, or -(CH2CH2O) n It consists of one, two, or three or more of the CH2CH2- groups, where n is 2, 3, 4, 5, 6, 7, or 8. Q a The compound described in any one of the following sections 61 to 113, where R is one, two or more of -C(=O)-, -NR-, -NR(C=O)-, and -(C=O)NR-, where R is methyl and / or hydrogen, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0212] 115.L a These are ethylene and / or -(CH2CH2O)4CH2CH2- or pentylene, Qa The compounds described in item 114, which are -C(=O)-, -N(CH3)-, -NH(C=O)-, -C(=O)NH-, or -C(=O)-N(CH3)-, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0213] 116.L b This is selected from C1-C6 linear or branched alkylenes, C2-C6 linear or branched alkenylenes, C2-C6 linear or branched alkynylenes, C6-C6 linear or branched alkynylenes, arylenes having one or more rings and 6-10 carbon atoms, or polyethylene glycol groups. The aforementioned polyethylene glycol group is -(CH2CH2O) n -, -O(CH2CH2O) n -,-(CH2CH2O) n CH2-, -(CH2CH2O) n CH2CH2-, -CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n It consists of one, two, or three or more CH2- groups, where n is an integer of 1 or more, and the alkylene, alkenylene, and alkynylene are optionally substituted by substituents. Q b is none, -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-, where R is selected from hydrogen, optionally substituted C1-C6 linear or branched alkyl, a compound described in any one of items 61-115, or its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0214] 117.L b This refers to C1-C6 straight-chain or branched-chain alkylenes, or C6-C610 It is one, two, or three or more of the allerenes. Q b is one, two or more of -C(=O)-, -C(=O)NR-, -OC(=O)NR-, -NRC(=O)-, -NRC(=O)O-, or -NRC(=O)NR-, where R is selected from hydrogen, optionally substituted C1-C6 linear or branched alkyl groups, the compounds described in item 116, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0215] 118.L b This is one, two, or three or more of methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, sec-butylene, tert-butylene, or bendiylene. Q b The compound described in item 116, or its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof, wherein R is none, -C(=O)-, -NRC(=O)-, -NRC(=O)NR-, or -C(=O)NR-, where R is hydrogen or methyl.

[0216] 119. The substituents are selected from deuterium, tritium, halogen atoms, amino, hydroxy, cyano, nitro, optionally substituted alkyl, alkenyl, alkynyl groups, optionally substituted cyclyl groups, acyl, optionally substituted amino, optionally substituted carbamoyl, optionally substituted thiocarbamoyl, optionally substituted sulfamoyl, optionally substituted hydroxy, optionally substituted thioalkyl (SH), and optionally substituted silyl, and are compounds described in any one of items 61 to 118, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0217] 120.j is an integer from 1 to 20, and is a compound described in any one of items 61 to 119, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0218] 121. The compound is a compound described in any one of the items 61 to 120, selected from the structures described below, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0219] [ka]

[0220] [ka]

[0221] [ka]

[0222] 122. The antibody is selected from the group consisting of mouse antibodies, chimeric antibodies, humanized antibodies, and fully human antibodies, and is a compound described in any one of items 61 to 121, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0223] 123. The antibody includes a monoclonal antibody, or a compound described in any one of items 61 to 122, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0224] 124. The antibody includes a bispecific antibody, or a compound described in any one of items 61 to 123, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0225] 125. The antigen-binding fragment is Fab, Fab ’ , Fv fragment, F(ab ’ Compounds described in any one of items 61 to 124, selected from the group consisting of )2, F(ab)2, scFv, di-scFv, and VHH, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0226] 126. Preface Ab , Anti-Crowden 18.2 antibody, Anti-Folic Acid 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 Cycase 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 antibody Protein 3 antibody, anti-Claudin 18.2 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 Compounds described in any one of items 61 to 125, selected from the group consisting of the anti-Ly6E antibody, anti-MFI2 antibody, anti-NOTCH3 antibody, anti-p-cadherin antibody, anti-PRLR antibody, anti-RNF43 antibody, and antigen-binding fragments of the above antibodies, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0227] 127. The heavy chains HCDR1, HCDR2, and HCDR3 of Ab and the light chains LCDR1, LCDR2, and LCDR3 are, respectively, compounds described in any one of the items 61 to 126, or tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0228] 128. The heavy chain variable region VH and light chain variable region VL of Ab are, respectively, compounds described in any one of items 61 to 127, or tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0229] 129. The heavy chain and light chain of Ab are, respectively, compounds described in any one of the items 61 to 128, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0230] 130. The Ab is an anti-claudin 18.2 antibody or an antigen-binding fragment thereof, one of the compounds described in any one of the items 61 to 129, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

[0231] 131. The Ab is an anti-human folate receptor antibody or an antigen-binding fragment thereof, one of the compounds described in any one of the items 61 to 130, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

[0232] 132. A pharmaceutical composition, A pharmaceutical composition comprising any compound described in any one of items 1 to 131, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0233] 133. Use of any compound described in any one of sections 1 to 132, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof, in the preparation of a drug for treating and / or prophylactic purposes of tumors.

[0234] 134. The aforementioned tumor contains 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 (delta-like protein 3), 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 Use as described in item 133, selected from tumors associated with the expression of targets from the group consisting of H, GPC3, LAMP-1, LRRC15, Ly6E, MFI2, NOTCH3, p-cadherin, PRLR, and RNF43.

[0235] 135. The use according to paragraph 133, wherein tumors associated with the expression of the target include tumors in which the target is highly expressed and / or tumors in which the target is positive.

[0236] 136. The use described in paragraph 133, wherein the tumor is selected from solid tumors, hematological malignancies, and metastatic, refractory, or recurrent lesions of cancer.

[0237] 137. The use described in paragraph 133, wherein the tumor is selected from the group consisting of esophageal cancer, gastrointestinal cancer, pancreatic cancer, thyroid cancer, colorectal cancer, kidney cancer, lung cancer (e.g., non-small cell lung cancer), liver cancer, stomach 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. [Effects of the Invention]

[0238] Compared to the anionic and cationic hydrophilic groups of PEG, the antibody-drug conjugates provided in this application, i.e., fluorophore-antibody-drug conjugates modified with water-soluble groups in the form of sulfonic acid endosal salts, significantly increase specific accumulation at tumor sites. The antibody-conjugates of this application have strongly hydrophilic sulfonic acid endosal salt and / or phosphate endosal salt side chains introduced into the polypeptide (VC, VA, AAA, etc.)-PAB (self-destructing group) site of the linker. Sulfonic acid groups or phosphate groups are strongly hydrophilic because they possess both hydrogen bond donors and hydrogen bond acceptors, and sulfonic acid endosal salts or phosphate endosal salts can further enhance hydrophilicity because they possess both positive and negative charges. Furthermore, water-soluble groups in the form of endosal salts may enhance tumor targeting of the antibody-drug conjugate and reduce nonspecific binding, thereby significantly improving the drugability (including hydrophilicity and stability) of the antibody-drug conjugate under test, reducing drug clearance in the body, and enhancing tumor therapeutic effects. [Brief explanation of the drawing]

[0239] The drawings are provided for the purpose of better understanding the present application and do not constitute an unwarranted limitation thereto. [Figure 1] These are size exclusion chromatography (SEC) analysis diagrams of the antibody-drug conjugates of Comparative Example 2 and Test Example 6 provided in this application. [Figure 2]These are size exclusion chromatography (SEC) analysis diagrams of the antibody-drug conjugates of Comparative Example 3 and Test Example 9 provided in this application. [Figure 3] These are size exclusion chromatography (SEC) analysis diagrams of the antibody-drug conjugates of Comparative Example 1 and Test Examples 1, 5a, 7, 9, and 11 provided in this application. [Figure 4] These are hydrophobic interaction chromatography (HIC) analysis diagrams of the antibody-drug conjugates of Comparative Example 2 and Test Example 6 provided in this application. [Figure 5] These are hydrophobic interaction chromatography (HIC) analysis diagrams of the antibody-drug conjugates of Comparative Example 3 and Test Example 9 provided in this application. [Figure 6] This is a chart of the hydrophobic interaction chromatography (HIC) analysis 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. [Figure 7] This is the light chain deconvolution mass spectrum of test example 5a provided in this application. [Figure 8] This is the heavy chain deconvolution mass spectrum of test example 5a provided in this application. [Figure 9] This is the reversed-phase chromatogram of test example 5a provided in this application. [Figure 10] This is an in vitro cell-killing curve for MC38-18.2 cells of the 9D1 antibody-drug conjugate provided in this application and a comparative sample. [Figure 11] This is the tumor therapeutic action curve of the 9D1 antibody-drug conjugate provided in this application for the MC38-hClaudin18.2 mouse subcutaneous tumor model. [Figure 12] This is the tumor therapeutic action curve of the 9D1 antibody-drug conjugate provided in this application for a NUGC4-hClaudin18.2 mouse subcutaneous tumor model. [Figure 13] This is the tumor therapeutic action curve of the Farletuzumab-LD-38 antibody-drug conjugate provided in this application for an OVCAR-3 mouse subcutaneous tumor model. [Figure 14]This is the tumor therapeutic action curve of the Farletuzumab-LD-38 antibody-drug conjugate provided in this application for the IGROV1 mouse subcutaneous tumor model. [Figure 15] These are the pharmacokinetic curves of 9D1-LD-38 and 9D1-GGFG-Exatecan provided in this application in rats. [Figure 16] This describes the killing activity of 9D1-LD-38, 9D1-LD-82, 9D1-LD-88, and 9D1-LD-89 against MC38-hClaudin18.2 cells. [Figure 17] This is the result of immunogenic cell death induced by 9D1-LD-38 and 9D1-GGFG-Dxd in B16F10-hClaudin18.2 cells. [Figure 18] This describes the bystander-killing effect of 9D1-LD-38 and 9D1-GGFG-Dxd on HEK293-hClaudin18.2 / HEK293 cells. [Figure 19] This result shows that the free toxin Exatecan is released after digestion of 9D1-LD-38. [Figure 20] These are the in vitro serum incubation stability results for 9D1-LD-38, 9D1-LD-65, 9D1-LD-73, 9D1-LD-74, and 9D1-Deruxtecan (9D1-GGFG-Dxd). [Figure 21] These are the results of changes in in vivo DAR values ​​for 9D1-LD-38, 9D1-LD-65, and 9D1-Deruxtecan (9D1-GGFG-Dxd). [Figure 22] These are the total toxin (bound toxin + free toxin) and free toxin content in the plasma of tumor-bearing mice administered 9D1-LD-38, 9D1-LD-65, and 9D1-Deruxtecan (9D1-GGFG-Dxd). [Figure 23] These are the total toxin (bound toxin + free toxin) and free toxin content in the tissues of tumor-bearing mice administered 9D1-LD-38, 9D1-LD-65, and 9D1-Deruxtecan (9D1-GGFG-Dxd). [Figure 24]These are the pharmacodynamic results for 9D1-LD-38, 9D1-Deruxtecan (9D1-GGFG-Dxd), and 9D1-vc-PAB-MMAE in p-gp-mediated drug-resistant tumor-bearing mouse models. [Figure 25] These are the pharmacodynamic results in a mouse model with normal immune function and tumors treated with ICI and 9D1-LD-38. [Figure 26] This shows the bystander-killing effect of 9D1-LD-38 on tumor-bearing mice. [Modes for carrying out the invention]

[0240] The following description will focus on exemplary embodiments of the present application, including various details of the embodiments for the sake of ease of understanding, which should be considered merely examples. Those skilled in the art will understand that various changes and modifications of the embodiments described herein can be made without departing from the scope and spirit of the present application. Furthermore, for clarity and brevity, well-known descriptions of functions and structures will be omitted in the following description.

[0241] Unless otherwise stated, some terms used in the specification and claims of this application are defined as follows:

[0242] The alkyl group may be branched or unbranched, and preferably has 1 to about 12 carbon atoms. More preferably, the alkyl group has 1 to about 6 carbon atoms. Even more preferably, it is an alkyl group with 1, 2, 3, or 4 carbon atoms. Examples of 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, and 2,3-dimethylbutyl. The alkyl group may be substituted or unsubstituted.

[0243] The "alkenyl" may be branched or unbranched and has one or more double bonds and 2 to about 12 carbon atoms. More preferred alkenyls have 2 to about 6 carbon atoms. Even more preferred are alkenyls with 2, 3, or 4 carbon atoms. Examples of alkenyl groups include, but are not limited to, vinyl, 1-propenyl, 2-propenyl, 1-methylvinyl, 1-butenyl, 2-butenyl, and 3-butenyl. The alkenyl may be substituted or unsubstituted.

[0244] "Alkynyl" may be branched or unbranched, and has one or more triple bonds and 2 to about 12 carbon atoms. More preferred alkynyl groups have 2 to 6 carbon atoms, and even more preferably alkynyl groups with 2, 3, or 4 carbon atoms. Examples of alkynyl groups include, but are not limited to, acetylene, propyne, and 1-butyne.

[0245] "Aliphatic cyclyl" refers to a cyclic substituent that does not possess aromaticity, and may be a saturated hydrocarbon substituent or an unsaturated hydrocarbon substituent, such as a cycloalkyl or cycloalkenyl substituent.

[0246] An "aliphatic heterocyclyl" refers to a substituent formed by a heterocyclic compound that has at least one heteroatom in its ring skeleton and is not aromatic. The heteroatom may be oxygen, nitrogen, sulfur, etc., and includes monocyclic, polycyclic, fused, bridging, and spirocyclic structures, such as a 5- to 7-membered monocyclic or a 7- to 10-membered dicyclic or tricyclic, which may contain one, two, or three atoms selected from nitrogen, oxygen, and / or sulfur. Aliphatic heterocyclyls include, but are not limited to, morpholinyl, oxetanyl, thiomorpholinyl, tetrahydrofuranyl, tetrahydropyranyl, 1,1-dioxo-thiomorpholinyl, piperidinyl, 2-oxo-piperidinyl, pyrrolidinyl, 2-oxo-pyrrolidinyl, piperazine-2-one, 8-oxa-3-azabicyclo[3.2.1]octyl, piperazinyl, and hexahydropyrimidine. The heterocyclic group may be substituted or unsubstituted.

[0247] "Aryl" refers to a carbocyclic aromatic system containing one or two rings, which may be fused and linked together. The term "aryl" includes monocyclic or bicyclic aromatic groups such as phenyl, naphthyl, and tetrahydronaphthyl aromatic groups. Preferred aryls are C6-C10 aryls, more preferred aryls are phenyl and naphthyl, and most preferred aryl is naphthyl. Aryls may be substituted or unsubstituted.

[0248] A "heteroaryl" refers to an aromatic 5-6 member monocyclic or 8-10 member bicyclic ring that can contain 1-4 atoms selected from nitrogen, oxygen, and / or sulfur. Specific examples of heteroaryls include, but are not limited to, furanyl, 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, benzoisothiazolyl, benzoxazolyl, and benzoisoxazolyl. Heteroaryls may be substituted or unsubstituted.

[0249] "Substituted" means that one or more hydrogen atoms in the group, preferably up to five, more preferably one to three, are substituted independently by a corresponding number of substituents. Needless to say, substituents are only in substitutable chemical positions, and those skilled in the art can determine possible or impossible substitutions (by experiment or theory) without excessive effort. For example, amino or hydroxyl groups with free hydrogen can become unstable when bonded to carbon atoms with unsaturated (e.g., olefin) bonds.

[0250] As used herein, “substituted” or “substituted” means that the group may be substituted with one or more groups selected from alkyl, alkoxy, alkylthio, alkylamino, halogen, thio, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, amino, haloalkyl, and hydroxyalkyl substituents, unless otherwise specified.

[0251] If a group is described as "optionally substituted by substituents," that group may be unsubstituted or substituted by one or more of the substituents listed. Similarly, if a group is described as "unsubstituted or substituted," and is substituted, the substituent may be one or more selected from the substituents listed. If no substituent is specified, a group that is "optionally substituted by substituents" or "substituted" is an alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, aliphatic heterocyclyl, aralkyl, heteroaralkyl, (aliphatic heterocyclyl)alkyl, hydroxy, protected hydroxy, alkoxy, aryloxy, acyl, mercapto, alkylthio, arylthio, cyano, halogen, thiocarbonyl, O-carbamoyl, N-carbamoyl, O-thiocarbamoyl This means that the group may be substituted with one or more groups individually or independently selected from N-thiocarbamoyl, C-amide, N-amide, S-sulfonylamino, N-sulfonylamino, C-carboxy, protected C-carboxy, O-carboxy, isocyanate group, thiocyanate salt, isothiocyanate group, nitro, silyl, thio, sulfinyl, sulfonyl, haloalkyl, haloalkoxy, trihalomethanesulfonyl, trihalomethanesulfonamide, amino, monosubstituted amino and disubstituted amino and their protected derivatives.

[0252] Halogen substituents include F, Cl, Br, and I. "Hydroxy" refers to the -OH group. "Halogen" refers to fluorine, chlorine, bromine, and iodine.

[0253] "Amino" refers to -NH2. "Cyano" refers to -CN. "Nitro" refers to -NO2. "Carboxy" refers to -C(O)OH. "Acyl" refers to -C(O)R.

[0254] The term "hydrogen" or "H" in chemical structures used herein, unless otherwise specified, 1 Not only H, but also deuterium ( 2H, D), tritium ( 3 It should be understood that this includes H, T), or mixtures thereof. "Deuterated isotope derivative" refers to the corresponding compound obtained when one or more H atoms in the compound are substituted with D atoms.

[0255] "Pharmaceutical composition" means a mixture comprising one or more of the compounds described herein or their physiologically pharmaceutically acceptable salts or prodrugs, other chemical components, and other components such as physiologically pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to facilitate administration to a living organism, promote the absorption of the active ingredient, and thereby exert biological activity.

[0256] This application proposes a compound represented by formula (1), or its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof. The structure of formula (1) is shown below.

[0257] [ka]

[0258] (M is a linking group, and Sp is -L) a -Q a - where A is selected from none, hydrogen, or alkyl, T represents a linear or branched (C1-C8) trivalent alkyl group, and Y, Y', and Y'' are -L respectively. b -Q b - and L a , L b Each is selected from none, alkylene, alkenylene, alkylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, heteroarylene, or polyethylene glycol group, where the alkylene, alkenylene, alkylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, heteroarylene, and polyethylene glycol groups are optionally substituted with substituents, Q a Q bEach is selected from none, -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-, where R is selected from hydrogen or optionally substituted alkyl, and Z is none. Z', Z'' are selected from hydrogen, optionally substituted alkyl, or sulfonic acid internal salts, with none, hydrogen, optionally substituted alkyl, sulfonic acid internal salt, or phosphate internal salt, and at least one of Z, Z', Z'' is a sulfonic acid internal salt or phosphate internal salt, X contains a self-destructing unit or none, D contains a drug molecule, i is an integer of 1 or more, 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, q are integers from 0 to 5, for example 0, 1, 2, 3, 4, 5, etc.

[0259] If h is greater than 1, then multiple Sps may have different structures or the same structure, i.e., multiple Ls a These may be different structures or identical structures, and there may be multiple Qs. a The structures may be different or identical. a and Q a The number of each can be the same or different.

[0260] If i is greater than 1, then 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.

[0261] If p is greater than 1, multiple Y's may have different structures or the same structure.

[0262] If q is greater than 1, multiple Y'' may have different structures or the same structure.

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

[0264] [ka]

[0265] (Here, the definitions of M, Sp, A, T, Y, h, o, i, X, D, and Z are as described above, and at least one of the i Zs is an intrasulfonic acid salt.)

[0266] In this application, formula (1-1) is formula (1-1-1).

[0267] [ka]

[0268] (Here, AA are amino acid residues, and w is an integer greater than or equal to 1, such as 1, 2, 3, 4, 5, 6, etc. If w is greater than 1, multiple AAs may be residues of the same amino acid or residues of different amino acids.)

[0269] In this application, formula (1) is formula (1-2).

[0270] [ka]

[0271] (Here, the definitions of M, Sp, A, T, Y, Z, o, p, i, h, X, Y', and D are as described above, and Z' is an endosodium sulfonic acid or endosodium phosphate.)

[0272] In this application, formula (1) is formula (1-2-1).

[0273] [ka]

[0274] (Here, AA are amino acid residues, and w is an integer greater than or equal to 1, such as 1, 2, 3, 4, 5, 6, etc. If w is greater than 1, multiple AAs may be residues of the same amino acid or residues of different amino acids.)

[0275] In this application, formula (1) is formula (1-3).

[0276] [ka]

[0277] (Here, the definitions of M, Sp, A, Y, T, Z, o, i, h, q, X, Y'', and D are as described above, and Z'' is an end salt of sulfonic acid or an end salt of phosphate.) In this application, if Z is hydrogen or an optionally substituted alkyl group,

[0278] [ka]

[0279] AA is an amino acid residue, also known as AA.

[0280] In this application, if Z is an internal sulfonic acid salt,

[0281] [ka]

[0282] teeth,

[0283] [ka]

[0284] That is the case. In this application, if both A and Z are absent, then N, Y, and T are rings.

[0285] [ka]

[0286] To form, that is,

[0287] [ka]

[0288] Preferably

[0289] [ka]

[0290] That is the case. In this application, when Z is selected from hydrogen or optionally substituted alkyl, formula (1) becomes formula (1-3-1).

[0291] [ka]

[0292] (Here, AA are amino acid residues, and w is an integer greater than or equal to 1, such as 1, 2, 3, 4, 5, 6, etc. If w is greater than 1, multiple AAs may be residues of the same amino acid or residues of different amino acids.)

[0293] In this application, the amino acid is one or more selected from alanine, arginine, asparagine, aspartic acid, γ-carboxyglutamic acid, citrulline, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, n-leucine, norvaline, ornithine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine.

[0294] In this application, the amino acid is one or more selected from valine, glutamic acid, citrulline, lysine, alanine, phenylalanine, arginine, glutamine, asparagine, and glycine.

[0295] In this application, the amino acid is one or more selected from valine, citrulline, alanine, phenylalanine, glutamine, and glycine.

[0296] When w is greater than 1, multiple AAs may be the same or different, specifically as follows:

[0297] In some embodiments, when w is 2, the two AAs may be, independently, a valine residue and a citrulline residue.

[0298] In some embodiments, when w is greater than 2, the w AAs may each be independently a valine residue and a citrulline residue, where the number of valine residues may be one or more, or the number of citrulline residues may be one or more. For example, when w is 3, the three AAs each consist of two valine residues and one citrulline residue. As another example, when w is 3, the three AAs each consist of one valine residue and two citrulline residues.

[0299] In some embodiments, when w is 2, the two AAs may be, independently, a valine residue and an alanine residue.

[0300] In some embodiments, when w is greater than 2, the w AAs may each be independently a valine residue and an alanine residue, where the number of valine residues may be one or more, or the number of alanine residues may be one or more. For example, when w is 3, the three AAs each consist of two valine residues and one alanine residue. As another example, when w is 3, the three AAs each consist of one valine residue and two alanine residues.

[0301] In some embodiments, when w is 2, the two AAs may be, independently, a valine residue and a phenylalanine residue.

[0302] In some embodiments, when w is greater than 2, the w AAs may each be independently a valine residue and a phenylalanine residue, where the number of valine residues may be one or more, or the number of phenylalanine residues may be one or more. For example, when w is 3, the three AAs each consist of two valine residues and one phenylalanine residue. As another example, when w is 3, the three AAs each consist of one valine residue and two phenylalanine residues.

[0303] In some embodiments, when w is 2, the two AAs may be, independently, a valine residue and a glutamine residue.

[0304] In some embodiments, when w is greater than 2, the w AAs may each be independently a valine residue and a glutamine residue, where the number of valine residues may be one or more, or the number of glutamine residues may be one or more. For example, when w is 3, the three AAs each consist of two valine residues and one glutamine residue. As another example, when w is 3, the three AAs each consist of one valine residue and two glutamine residues.

[0305] In some embodiments, when w is 2, the two AAs may be a valine residue and a glycine residue, respectively.

[0306] In some embodiments, when w is greater than 2, the w AAs may each be independently a valine residue and a glycine residue, where the number of valine residues may be one or more, or the number of glycine residues may be one or more. For example, when w is 3, the three AAs each consist of two valine residues and one glycine residue. As another example, when w is 3, the three AAs each consist of one valine residue and two glycine residues.

[0307] In some embodiments, when w is 2, the two AAs may be, independently, a citrulline residue and an alanine residue.

[0308] In some embodiments, when w is greater than 2, the w AAs may each be independently a citrulline residue and an alanine residue, where the number of citrulline residues may be one or more, or the number of alanine residues may be one or more. For example, when w is 3, the three AAs each consist of two citrulline residues and one alanine residue. As another example, when w is 3, the three AAs each consist of one citrulline residue and two alanine residues.

[0309] In some embodiments, when w is greater than 1, the w AAs may each be independently a citrulline residue and a phenylalanine residue, where the number of citrulline residues may be one or more, or the number of phenylalanine residues may be one or more.

[0310] In some embodiments, when w is greater than 1, the w AAs may each be independently a citrulline residue and a glutamine residue, where the number of citrulline residues may be one or more, or the number of glutamine residues may be one or more.

[0311] In some embodiments, when w is greater than 1, the w AAs may each be independently a citrulline residue and a glycine residue, where the number of citrulline residues may be one or more, or the number of glycine residues may be one or more.

[0312] In some embodiments, when w is greater than 1, the w AAs may each be independently an alanine residue and a phenylalanine residue, where the number of alanine residues may be one or more, or the number of phenylalanine residues may be one or more.

[0313] In some embodiments, when w is greater than 1, the w AAs may each be independently an alanine residue and a glutamine residue, where the number of alanine residues may be one or more, or the number of glutamine residues may be one or more.

[0314] In some embodiments, when w is greater than 1, the w AAs may each be independently an alanine residue and a glycine residue, where the number of alanine residues may be one or more, or the number of glycine residues may be one or more.

[0315] In some embodiments, when w is greater than 1, the w AAs may each be independently a phenylalanine residue and a glutamine residue, where the number of phenylalanine residues may be one or more, or the number of glutamine residues may be one or more.

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

[0317] In some embodiments, when w is greater than 1, the w AAs may each be independently a glutamine residue and a glycine residue, where the number of glutamine residues may be one or more, or the number of glycine residues may be one or more.

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

[0319] In some embodiments, when w is greater than 1, the w AAs may each be independently a valine residue, a citrulline residue, and a phenylalanine residue, where the number of valine residues may be one or more, the number of citrulline residues may be one or more, or the number of phenylalanine residues may be one or more.

[0320] In some embodiments, when w is greater than 1, the w AAs may each be independently a valine residue, a citrulline residue, and a glutamine residue, where the number of valine residues may be one or more, the number of citrulline residues may be one or more, or the number of glutamine residues may be one or more.

[0321] In some embodiments, when w is greater than 1, the w AAs may each be independently a valine residue, a citrulline residue, and a glycine residue, where the number of valine residues may be one or more, the number of citrulline residues may be one or more, or the number of glycine residues may be one or more.

[0322] In some embodiments, when w is greater than 1, the w AAs may each be independently a valine residue, an alanine residue, and a phenylalanine residue, where the number of valine residues may be one or more, the number of alanine residues may be one or more, or the number of phenylalanine residues may be one or more.

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

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

[0325] In some embodiments, when w is greater than 1, the w AAs may each be independently a valine residue, a phenylalanine residue, and a glutamine residue, where the number of valine residues may be one or more, the number of phenylalanine residues may be one or more, or the number of glutamine residues may be one or more.

[0326] In some embodiments, when w is greater than 1, the w AAs may each be independently a valine residue, a phenylalanine residue, and a glycine residue, where the number of valine residues may be one or more, the number of phenylalanine residues may be one or more, or the number of glycine residues may be one or more.

[0327] In some embodiments, when w is greater than 1, the w AAs may each be independently a valine residue, a glutamine residue, and a glycine residue, where the number of valine residues may be one or more, the number of glutamine residues may be one or more, or the number of glycine residues may be one or more.

[0328] In some embodiments, when w is greater than 1, the w AAs may each be independently a citrulline residue, an alanine residue, and a phenylalanine residue, where the number of citrulline residues may be one or more, the number of alanine residues may be one or more, or the number of phenylalanine residues may be one or more.

[0329] In some embodiments, when w is greater than 1, the w AAs may each be independently a citrulline residue, an alanine residue, and a glutamine residue, where the number of citrulline residues may be one or more, the number of alanine residues may be one or more, or the number of glutamine residues may be one or more.

[0330] In some embodiments, when w is greater than 1, the w AAs may each be independently a citrulline residue, an alanine residue, and a glycine residue, where the number of citrulline residues may be one or more, the number of alanine residues may be one or more, or the number of glycine residues may be one or more.

[0331] In some embodiments, when w is greater than 1, the w AAs may each be independently an alanine residue, a phenylalanine residue, and a glutamine residue, where the number of alanine residues may be one or more, the number of phenylalanine residues may be one or more, or the number of glutamine residues may be one or more.

[0332] In some embodiments, when w is greater than 1, the w AAs may each be independently an alanine residue, a phenylalanine residue, and a glycine residue, where the number of alanine residues may be one or more, the number of phenylalanine residues may be one or more, or the number of glycine residues may be one or more.

[0333] In some embodiments, when w is greater than 1, the w AAs may each be independently a phenylalanine residue, a glutamine residue, and a glycine residue, where the number of phenylalanine residues may be one or more, the number of glutamine residues may be one or more, or the number of glycine residues may be one or more.

[0334] In some embodiments, when w is greater than 1, the w AAs may each be independently a valine residue, a citrulline residue, an alanine residue, and a phenylalanine residue, where the number of valine residues may be one or more, the number of citrulline residues may be one or more, the number of alanine residues may be one or more, or the number of phenylalanine residues may be one or more.

[0335] In some embodiments, when w is greater than 1, the w AAs may each be independently a valine residue, a citrulline residue, an alanine residue, and a glutamine residue, where the number of valine residues may be one or more, the number of citrulline residues may be one or more, the number of alanine residues may be one or more, or the number of glutamine residues may be one or more.

[0336] In some embodiments, when w is greater than 1, the w AAs may each be independently a valine residue, a citrulline residue, an alanine residue, and a glycine residue, where the number of valine residues may be one or more, the number of citrulline residues may be one or more, the number of alanine residues may be one or more, or the number of glycine residues may be one or more.

[0337] In some embodiments, when w is greater than 1, the w AAs may each be independently a citrulline residue, an alanine residue, a phenylalanine residue, and a glutamine residue, where the number of citrulline residues may be one or more, the number of alanine residues may be one or more, the number of phenylalanine residues may be one or more, or the number of glutamine residues may be one or more.

[0338] In some embodiments, when w is greater than 1, the w AAs may each be independently a citrulline residue, an alanine residue, a phenylalanine residue, and a glycine residue, where the number of citrulline residues may be one or more, the number of alanine residues may be one or more, the number of phenylalanine residues may be one or more, or the number of glycine residues may be one or more.

[0339] In some embodiments, when w is greater than 1, the w AAs may each be independently an alanine residue, a phenylalanine residue, a glutamine residue, and a glycine residue, where the number of alanine residues may be one or more, the number of phenylalanine residues may be one or more, the number of glutamine residues may be one or more, or the number of glycine residues may be one or more.

[0340] In some embodiments, when w is greater than 1, the w AAs may each be independently a valine residue, a citrulline residue, an alanine residue, a phenylalanine residue, and a glutamine residue, where the number of valine residues may be one or more, the number of citrulline residues may be one or more, the number of alanine residues may be one or more, the number of phenylalanine residues may be one or more, or the number of glutamine residues may be one or more.

[0341] In some embodiments, when w is greater than 1, the w AAs may each be independently a valine residue, a citrulline residue, an alanine residue, a phenylalanine residue, and a glycine residue, where the number of valine residues may be one or more, the number of citrulline residues may be one or more, the number of alanine residues may be one or more, the number of phenylalanine residues may be one or more, or the number of glycine residues may be one or more.

[0342] In some embodiments, when w is greater than 1, the w AAs may each be independently a citrulline residue, an alanine residue, a phenylalanine residue, a glutamine residue, and a glycine residue, where the number of citrulline residues may be one or more, the number of alanine residues may be one or more, the number of phenylalanine residues may be one or more, the number of glutamine residues may be one or more, or the number of glycine residues may be one or more.

[0343] In this application, M is N3, SH, ONH2, NH2, CO2H, (alkyl)-CO-, HCO-, BrCH 2- ICH2-,

[0344] [ka]

[0345] That is the case. Furthermore, M said,

[0346] [ka]

[0347] That is the case. In some embodiments, M is

[0348] [ka]

[0349] That is the case. In some embodiments, M is

[0350] [ka]

[0351] That is the case. In some embodiments, M is

[0352] [ka]

[0353] That is the case. In some embodiments, M is

[0354] [ka]

[0355] That is the case. In this application, A is hydrogen, methyl, ethyl, or propyl.

[0356] In some embodiments, A is hydrogen. In some embodiments, A is methyl. In some embodiments, A is ethyl. In some embodiments, A is propyl.

[0357] In this application, T is one of the following structures.

[0358] [ka]

[0359] In some embodiments, T is

[0360] [ka]

[0361] That is the case. In this application, L a The group consists of one, two, or three or more elements selected from alkylene, alkenylene, alkynylene, saturated or unsaturated aliphatic cyclylene with 3 to 10 ring carbon atoms, saturated or unsaturated aliphatic heterocyclylene with 3 to 10 ring carbon atoms and 1 to 4 heteroatoms, arylene with 6 to 18 carbon atoms having one or more rings, heteroarylene with 6 to 18 carbon atoms having one or more rings and 1 to 4 heteroatoms, or polyethylene glycol groups. The alkylene, alkenylene, alkynylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, and heteroarylene are optionally substituted with substituents, and the heteroatoms are selected from nitrogen, oxygen, or sulfur.

[0362] Furthermore, L a C1~C 10 Straight-chain or branched-chain alkylenes, C2-C 12 Straight-chain or branched-chain alkenylenes, C2-C 12 One, two, or three or more selected from linear or branched alkylenes, saturated or unsaturated aliphatic cyclylenes with 3 to 6 ring carbon atoms, saturated or unsaturated aliphatic heterocyclylenes containing heteroatoms and having 3 to 6 ring carbon atoms and 1 to 4 heteroatoms, arylenes with 6 to 10 carbon atoms having one or more rings, heteroarylenes with 6 to 10 carbon atoms having one or more rings and 1 to 4 heteroatoms, or polyethylene glycol groups, wherein the alkylenes, alkenylenes, alkylenes, aliphatic cyclylenes, aliphatic heterocyclylenes, arylenes, and heteroarylenes are optionally substituted with substituents, and the heteroatoms are selected from nitrogen, oxygen, or sulfur. Furthermore, L aThis group consists of one, two, or three or more C1-C6 linear or branched alkylenes, C2-C6 linear or branched alkenylenes, C2-C6 linear or branched alkynylenes, or polyethylene glycol groups, and the alkylenes, alkenylenes, and alkynylenes are optionally substituted with substituents.

[0363] Furthermore, L a This is one, two, or three or more C1-C6 linear or branched alkylenes, or polyethylene glycol groups, wherein the polyethylene glycol groups are -(CH2CH2O) n -, -O(CH2CH2O) n -,-(CH2CH2O) n CH2-, -(CH2CH2O) n CH2CH2-, -CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n CH2-, where n is an integer greater than or equal to 1.

[0364] Furthermore, L a This refers to ethylene, linear propylene, linear butylene, linear pentylene, or -(CH2CH2O) n One, two, or three or more CH2CH2- groups, where n is 2, 3, 4, 5, 6, 7, or 8.

[0365] Furthermore, L a These are ethylene and / or -(CH2CH2O)4CH2CH2- or pentylene.

[0366] In some embodiments, L a There are two of them, one of which is ethylene and the other is -(CH2CH2O)4CH2CH2-.

[0367] In some embodiments, L a If the number is 1, it is ethylene. In some embodiments, L a If the number is 1, it is pentylene. In some embodiments, La If the number is 1, it is -(CH2CH2O)4CH2CH2-.

[0368] In this application, L b This is one, two, or three or more selected from alkylene, alkenylene, alkynylene, saturated or unsaturated aliphatic cyclylene with 3 to 10 ring carbon atoms, saturated or unsaturated aliphatic heterocyclylene with 3 to 10 ring carbon atoms and 1 to 4 heteroatoms, arylene with 6 to 18 carbon atoms having one or more rings, heteroarylene with 6 to 18 carbon atoms having one or more rings and 1 to 4 heteroatoms, or polyethylene glycol. The alkylene, alkenylene, alkynylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, and heteroarylene are optionally substituted with substituents, the heteroatoms being selected from nitrogen, oxygen, or sulfur.

[0369] Furthermore, L b C1~C 10 Straight-chain or branched-chain alkylenes, C2-C 12 Straight-chain or branched-chain alkenylenes, C2-C 12 One, two, or three or more selected from linear or branched alkynylenes, saturated or unsaturated aliphatic cyclylenes with 3 to 6 ring carbon atoms, saturated or unsaturated aliphatic heterocyclylenes containing heteroatoms and having 3 to 6 ring carbon atoms and 1 to 4 heteroatoms, arylenes with 6 to 10 carbon atoms having one or more rings, heteroarylenes with 6 to 10 carbon atoms and 1 to 4 heteroatoms having one or more rings, or polyethylene glycol. The alkylene, alkenylene, alkynylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, and heteroarylene are optionally substituted with substituents, the heteroatoms being selected from nitrogen, oxygen, or sulfur.

[0370] Furthermore, L bThis is selected from C1-C6 linear or branched alkylene, C2-C6 linear or branched alkenylene, C2-C6 linear or branched alkylylene, or polyethylene glycol group. The aforementioned polyethylene glycol group is -(CH2CH2O) n -, -O(CH2CH2O) n -,-(CH2CH2O) n CH2-, -(CH2CH2O) n CH2CH2-, -CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n The molecule consists of one, two, or three or more CH2- groups, where n is an integer of 1 or more, and the alkylene, alkenylene, and alkynylene are optionally substituted by substituents.

[0371] Furthermore, L b This refers to C1-C6 straight-chain or branched-chain alkylenes, or C6-C6 10 It is one, two, or three or more of the allirenes.

[0372] Furthermore, L b is one, two, or three or more of methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, sec-butylene, tert-butylene, or bendiylene.

[0373] L b If there are multiple L b These may be identical or different structures, and specifically, they are shown as follows:

[0374] In some embodiments, L b If the number is one or more, L b These are all methylene. In some embodiments, L b If the number is one or more, L b These are all ethylene. In some embodiments, L b If the number is one or more, L ball are n-propylene. In some embodiments, L b If the number is one or more, L b all are isopropylene. In some embodiments, L b If the number is one or more, L b These are all n-butylene. In some embodiments, L b If the number is one or more, L b all are isobutylene. In some embodiments, L b If the number is one or more, L b These are all sec-butylene. In some embodiments, L b If the number is one or more, L b These are all tert-butylene. In some embodiments, L b If the number is one or more, L b They are all benzylene.

[0375] In some embodiments, L b If there are multiple L b These are independently methylene and ethylene, where the number of methylenes may be one or more, and the number of ethylenes may be one or more, for example L b If the number is 3, then there are 3 L b These are two methylene molecules and one ethylene molecule, respectively. As another example, L b If the number is 3, then there are 3 L b These are one methylene molecule and two ethylene molecules, respectively.

[0376] In some embodiments, L b If there are multiple L b These are independently methylene and n-propylene, where the number of methylene atoms may be one or more, and the number of n-propylene atoms may be one or more, for example L b If the number is 3, then there are 3 L bThese are two methylene groups and one n-propylene group, respectively. As another example, L b If the number is 3, then there are 3 L b These consist of one methylene group and two n-propylene groups, respectively.

[0377] In some embodiments, L b If the number is greater than 1, multiple L b These are independently methylene and isopropylene, where the number of methylene atoms may be one or more, and the number of isopropylene atoms may be one or more.

[0378] In some embodiments, L b If the number is greater than 1, multiple L b These are independently methylene and n-butylene, where the number of methylene atoms may be one or more, and the number of n-butylene atoms may be one or more.

[0379] In some embodiments, L b If the number is greater than 1, multiple L b These are independently methylene and isobutylene, where the number of methylene atoms may be one or more, and the number of isobutylene atoms may be one or more.

[0380] In some embodiments, L b If the number is greater than 1, multiple L b These are independently methylene and sec-butylene, where the number of methylene may be one or more, and the number of sec-butylene may be one or more.

[0381] In some embodiments, L b If the number is greater than 1, multiple L b These are independently methylene and tert-butylene, where the number of methylene may be one or more, and the number of tert-butylene may be one or more.

[0382] In some embodiments, L b If the number is greater than 1, multiple L b These are independently methylene and bendiylene, where the number of methylene may be one or more, and the number of bendiylene may be one or more.

[0383] In some embodiments, L b If the number is greater than 1, multiple L b These are independently ethylene and n-propylene, where the number of ethylenes may be one or more, and the number of n-propylenes may be one or more.

[0384] In some embodiments, L b If the number is greater than 1, multiple L b These are independently ethylene and isopropylene, where the number of ethylenes may be one or more, and the number of isopropylenes may be one or more.

[0385] In some embodiments, L b If the number is greater than 1, multiple L b These are independently ethylene and n-butylene, where the number of ethylenes may be one or more, and the number of n-butylenes may be one or more.

[0386] In some embodiments, L b If the number is greater than 1, multiple L b These are independently ethylene and isobutylene, where the number of ethylenes may be one or more, and the number of isobutylenes may be one or more.

[0387] In some embodiments, L b If the number is greater than 1, multiple L bThese are independently ethylene and sec-butylene, where the number of ethylenes may be one or more, and the number of sec-butylenes may be one or more.

[0388] In some embodiments, L b If the number is greater than 1, multiple L b These are independently ethylene and tert-butylene, where the number of ethylenes may be one or more, and the number of tert-butylenes may be one or more.

[0389] In some embodiments, L b If the number is greater than 1, multiple L b These are independently ethylene and benzylene, where the number of ethylenes may be one or more, and the number of benzylenes may be one or more.

[0390] In some embodiments, L b If the number is greater than 1, multiple L b These are independently n-propylene and isopropylene, where the number of n-propylene atoms may be one or more, and the number of isopropylene atoms may be one or more.

[0391] In some embodiments, L b If the number is greater than 1, multiple L b These are independently n-propylene and n-butylene, where the number of n-propylene may be one or more, and the number of n-butylene may be one or more.

[0392] In some embodiments, L b If the number is greater than 1, multiple L b These are independently n-propylene and isobutylene, where the number of n-propylene may be one or more, and the number of isobutylene may be one or more.

[0393] In some embodiments, L b If the number is greater than 1, multiple L b These are independently n-propylene and sec-butylene, where the number of n-propylene may be one or more, and the number of sec-butylene may be one or more.

[0394] In some embodiments, L b If the number is greater than 1, multiple L b These are independently n-propylene and tert-butylene, where the number of n-propylene may be one or more, and the number of tert-butylene may be one or more.

[0395] In some embodiments, L b If the number is greater than 1, multiple L b These are independently n-propylene and bendiylene, where the number of n-propylene may be one or more, and the number of bendiylene may be one or more.

[0396] In some embodiments, L b If the number is greater than 1, multiple L b These are independently isopropylene and n-butylene, where the number of isopropylene may be one or more, and the number of n-butylene may be one or more.

[0397] In some embodiments, L b If the number is greater than 1, multiple L b These are independently isopropylene and isobutylene, where the number of isopropylene may be one or more, and the number of isobutylene may be one or more.

[0398] In some embodiments, L b If the number is greater than 1, multiple L bThese are independently isopropylene and sec-butylene, where the number of isopropylene may be one or more, and the number of sec-butylene may be one or more.

[0399] In some embodiments, L b If the number is greater than 1, multiple L b These are independently isopropylene and tert-butylene, where the number of isopropylene may be one or more, and the number of tert-butylene may be one or more.

[0400] In some embodiments, L b If the number is greater than 1, multiple L b These are independently isopropylene and benzylene, where the number of isopropylene may be one or more, and the number of benzylene may be one or more.

[0401] In some embodiments, L b If the number is greater than 1, multiple L b These are independently methylene, ethylene, and n-propylene, where the number of methylenes may be one or more, the number of ethylenes may be one or more, and the number of n-propylenes may be one or more.

[0402] In some embodiments, L b If the number is greater than 1, multiple L b These are independently methylene, ethylene, and isopropylene, where the number of methylenes may be one or more, the number of ethylenes may be one or more, and the number of isopropylenes may be one or more.

[0403] In some embodiments, L b If the number is greater than 1, multiple L bThese are independently methylene, ethylene, n-propylene, and isopropylene, where the number of methylenes may be one or more, the number of ethylenes may be one or more, the number of n-propylenes may be one or more, and the number of isopropylenes may be one or more.

[0404] In some embodiments, L b If the number is greater than 1, multiple L b These are independently methylene, ethylene, n-propylene, isopropylene, and n-butylene, where the number of methylenes may be one or more, the number of ethylenes may be one or more, the number of n-propylenes may be one or more, the number of isopropylenes may be one or more, and the number of n-butylenes may be one or more.

[0405] In this application, Q a R is selected from one, two, or three or more elements 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)-, and -OC(=O)O-, where R is selected from hydrogen and optionally substituted C1-C6 linear or branched alkyl groups.

[0406] Furthermore, Q a R is one, two, or three or more of -C(=O)-, -NR-, -C(=O)NR-, -NRC(=O)NR-, -OC(=O)O-, -OC(=O)NR-, -NRC(=O)-, or -OC(=O)-, where R is selected from hydrogen or optionally substituted C1-C6 linear or branched alkyl groups.

[0407] Furthermore, Q aR is one, two, or three or more of -C(=O)-, -NR-, -NRC(=O)-, and -(C=O)NR-, where R is selected from hydrogen or optionally substituted C1-C6 linear or branched alkyl groups.

[0408] Furthermore, Q a are -C(=O)-, -N(CH3)-, -NHC(=O)-, -C(=O)-NH-, -C(=O)-N(CH3)-.

[0409] In some embodiments, Q a If the number is 1, then it is -C(=O)-.

[0410] In some embodiments, Q a If there are two of these, one is -C(=O)- and the other is -C(=O)NH-.

[0411] In some embodiments, Q a If there are two of them, one is -C(=O)- and the other is -C(=O)-NR-, where R is methyl.

[0412] In some embodiments, Q a If there are two of them, one is -C(=O)- and the other is -NR-, where R is methyl.

[0413] In this application, Q b R is selected from none, -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-, where R is selected from hydrogen, or optionally substituted C1-C6 linear or branched alkyl groups.

[0414] Furthermore, Q bR is one, two or more of the following: none, -C(=O)NR-, -OC(=O)NR-, -NRC(=O)-, -NRC(=O)O-, or -NRC(=O)NR-, where R is selected from hydrogen, or optionally substituted C1-C6 linear or branched alkyl groups.

[0415] Furthermore, Q b R is one, two or more of the following: none, -C(=O)-, -NRC(=O)-, -NRC(=O)NR-, or -C(=O)NR-, where R is hydrogen or methyl.

[0416] In some embodiments, Q b There is none. In some embodiments, Q b is -C(=O)-. In some embodiments, Q b is -NRC(=O)-, and R is hydrogen. In some embodiments, Q b is -NRC(=O)-, and R is methyl. In some embodiments, Q b is -NRC(=O)NR-, where R is hydrogen. In some embodiments, Q b is -NRC(=O)NR-, where R is methyl. In some embodiments, Q b is -C(=O)NR-, where R is hydrogen. In some embodiments, Q b is -C(=O)NR-, where R is methyl. In this application, X is selected from the following:

[0417] [ka]

[0418] (The wavy line on the right in X is connected to D, R4 is hydrogen, C1-C6 straight or branched alkyl, -(CH2CH2O) m Selected from R2, -CH2CH2NR2R3, and -CH2CH2SO2R2, R2 and R3 are selected from hydrogen and a C1-C6 straight-chain or branched-chain alkyl group, respectively, and m is an integer greater than 1.

[0419] Furthermore, X said,

[0420] [ka]

[0421] And, R4 can be hydrogen, methyl, -CH2CH2OCH2CH2OH, -CH2CH2NMe2, or -CH2CH2SO2Me.

[0422] In some embodiments, X is

[0423] [ka]

[0424] Therefore, R4 is hydrogen. In some embodiments, X is

[0425] [ka]

[0426] Therefore, R4 is methyl. In some embodiments, X is

[0427] [ka]

[0428] Therefore, R4 is -CH2CH2OCH2CH2OH. In some embodiments, X is

[0429] [ka]

[0430] Therefore, R4 is -CH2CH2NMe2. In some embodiments, X is

[0431] [ka]

[0432] Therefore, R4 is -CH2CH2SO2Me. In some embodiments, X is

[0433] [ka]

[0434] Therefore, R4 is hydrogen. In some embodiments, X is

[0435] [ka]

[0436] Therefore, R4 is methyl. In some embodiments, X is

[0437] [ka]

[0438] Therefore, R4 is -CH2CH2OCH2CH2OH. In some embodiments, X is

[0439] [ka]

[0440] Therefore, R4 is -CH2CH2NMe2. In some embodiments, X is

[0441] [ka]

[0442] Therefore, R4 is -CH2CH2SO2Me. In some embodiments, X is

[0443] [ka]

[0444] Therefore, R4 is hydrogen. In some embodiments, X is

[0445] [ka]

[0446] Therefore, R4 is methyl. In some embodiments, X is

[0447] [ka]

[0448] Therefore, R4 is -CH2CH2OCH2CH2OH. In some embodiments, X is

[0449] [ka]

[0450] Therefore, R4 is -CH2CH2NMe2. In some embodiments, X is

[0451] [ka]

[0452] Therefore, R4 is -CH2CH2SO2Me.

[0453] In this application, the sulfonic acid salt is as follows:

[0454] [ka]

[0455] (The wavy line is connected to Y, Y', or Y'', Here, c, d, e, f, g are all integers of 1, 2, 3, 4, 5, or 6, and R6, R7, R 11 , R 12 , R 13 , R 14 (Each element is selected from none, hydrogen, or a C1-C6 linear or branched alkyl group.)

[0456] Furthermore, the sulfonic acid salt is as follows:

[0457] [ka]

[0458] (Here, R6, R7, R 11 , R 12 , R 13 and R 14 (Each is selected from none, methyl, or ethyl, and c, d, e, f, and g are 1, 2, or 3, respectively.)

[0459] Furthermore, the sulfonic acid salt is one of the following structures.

[0460] [ka]

[0461] In some embodiments, the sulfonic acid salt is

[0462] [ka]

[0463] That is the case. In some embodiments, the sulfonic acid salt is

[0464] [ka]

[0465] That is the case. In some embodiments, the sulfonic acid salt is

[0466] [ka]

[0467] That is the case. In some embodiments, the sulfonic acid salt is

[0468] [ka]

[0469] That is the case. In this application, the phosphate salt is as follows:

[0470] [ka]

[0471] (where a and b are all integers of 1, 2, 3, 4, 5 or 6, and R8, R9, and R 10 (Each element is selected from hydrogen, a C1-C6 linear chain, or a branched-chain alkyl group.)

[0472] Furthermore, the phosphate salts are as follows:

[0473] [ka]

[0474] (Here, R8, R9, R 10 (wherein 1 is methyl or ethyl, and 'a' is 1, 2, or 3, and 'b' is 1, 2, or 3.)

[0475] Furthermore, the phosphate salts are as follows:

[0476] [ka]

[0477] (Here, R8, R9, R 10 (All are methyl, a is 2, and b is 1.)

[0478] In this application, D includes a cytotoxin or an immunoagonist.

[0479] In this application, the cytotoxin is one or two selected from tubulin inhibitors, DNA synthesis inhibitors, topoisomerase inhibitors, or RNA polymerase 2 inhibitors.

[0480] In this application, the cytotoxin is one or more selected from auristatins, meitansinoid DMs, calicheamicin, duocarmycin, pyrrolobenzodiazepines, camptothecin derivatives, and α-amanitin.

[0481] In this application, the immunoagonist is selected from a TLR7 / 8 agonist or a STING agonist.

[0482] In this application, i is an integer between 2 and 8, and may be, for example, 2, 3, 4, 5, 6, 7, or 8.

[0483] In this application, h is an integer between 1 and 4, and may be, for example, 1, 2, 3, or 4.

[0484] In this application, o, p, and q are each integers from 0 to 3, and may be, for example, 0, 1, 2, or 3.

[0485] In this application, L a This is one, two, or three or more C1-C6 linear or branched alkylenes, or polyethylene glycol groups, wherein the polyethylene glycol groups are -(CH2CH2O) n -, -O(CH2CH2O) n -,-(CH2CH2O) n CH2-, -(CH2CH2O) n CH2CH2-, -CH2O(CH2CH2O) n -, or -CH2O(CH2CH2O) n CH2-, where n is an integer greater than or equal to 1, Q a R is one, two, or three or more of -C(=O)-, -NR(C=O)-, and -(C=O)NR-, where R is selected from hydrogen, C1-C6 linear, or branched alkyl groups.

[0486] Furthermore, L a This refers to ethylene, linear propylene, linear butylene, linear pentylene, or -(CH2CH2O) n It consists of one, two, or three or more of the CH2CH2- groups, where n is 2, 3, 4, 5, 6, 7, or 8, and Q a R is one, two, or three or more of -C(=O)-, -NR(C=O)-, and -(C=O)NR-, where R is methyl and / or hydrogen.

[0487] Furthermore, L a Q is ethylene and / or -(CH2CH2O)4CH2CH2- or pentylene, a These are -C(=O)-, -NR-, -NH(C=O)- and / or -C(=O)NH-.

[0488] In some embodiments, L a There are two of them, one of which is ethylene and the other is -(CH2CH2O)4CH2CH2-. Q a There are two such pairs, one of which is -C(=O)- and the other is -C(=O)NH-.

[0489] In some embodiments, L a The number is 1, and it is pentylene, Q a The number is one, and it is -C(=O)-.

[0490] In some embodiments, L a The number is 1, and it is -(CH2CH2O)4CH2CH2-, Q a There are two such pairs: one is -C(=O)- and the other is -C(=O)NH-.

[0491] In some embodiments, L a The number is 1, and it is -(CH2CH2O)4CH2CH2-, Q a There are two such pairs: one is -C(=O)- and the other is -C(=O)N(CH3)-.

[0492] In some embodiments, L a The number is two, and they are ethylene and pentylene, respectively, Q a There are two such entities: one is -C(=O)- and the other is -N(CH3)-.

[0493] In this application, L b This is selected from C1-C6 linear or branched alkylene, C2-C6 linear or branched alkenylene, C2-C6 linear or branched alkynylene, C6-C6 linear or branched arylene having one or more rings and having 6-10 carbon atoms, 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 It consists of one, two, or three or more CH2- groups, where n is an integer of 1 or more, and the alkylene, alkenylene, and alkynylene are optionally substituted by substituents. Q b R is selected from none, -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, or optionally substituted C1-C6 linear or branched alkyl groups.

[0494] Furthermore, L b This refers to C1-C6 straight-chain or branched-chain alkylenes, or C6-C6 10 One, two, or three or more of the allerenes, Q b R is one, two or more of the following: none, -C(=O)NR-, -C(=O)-, -OC(=O)NR-, -NRC(=O)-, -NRC(=O)O-, or -NRC(=O)NR-, where R is selected from hydrogen, C1-C6 linear or branched alkyl groups.

[0495] Furthermore, L b This is one, two, or three or more of methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, sec-butylene, tert-butylene, or bendiylene. Q b R is one, two or more of the following: none, -C(=O)-, -NRC(=O)-, -NRC(=O)NR-, or -C(=O)NR-, where R is hydrogen or methyl.

[0496] In some embodiments, in Y, L b If there are two of them, one is methylene and the other is isopropylene, and Q b There is none.

[0497] In some embodiments, in Y, L b If there are three of them, they are methylene, isopropylene, and ethylene, respectively, and Q b The number of is 1, which is -C(=O)NR-, and R is hydrogen.

[0498] In some embodiments, in Y, L b If there are two of them, one is ethylene and the other is isopropylene, and Q b The number of is 1, which is -C(=O)NR-, and R is hydrogen.

[0499] In some embodiments, in Y, L b If there are two of them, one is n-propylene and the other is isopropylene, and Q b The number is one, and it is -NRC(=O)NR-, where R is hydrogen.

[0500] In some embodiments, in Y, L b If the number is 1, then it is benzilene, and Q b There is none.

[0501] In some embodiments, in Y, L b If there are two of them, they are ethylene and isopropylene, respectively, and Q b The number is one, and it is -C(=O)-.

[0502] In this application, the substituent is selected from deuterium, tritium, halogen atoms, amino, hydroxy, cyano, nitro, optionally substituted alkyl, alkenyl, alkynyl groups, optionally substituted cyclyl groups, acyl, optionally substituted amino, optionally substituted carbamoyl, optionally substituted thiocarbamoyl, optionally substituted sulfamoyl, optionally substituted hydroxy, optionally substituted thioalkyl (SH), and optionally substituted silyl.

[0503] In this application, the compound is selected from the following structures.

[0504] [ka]

[0505] [ka]

[0506] [ka]

[0507] [ka]

[0508] The present invention also provides a compound represented by formula (2), or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof, the structure of formula (2) is shown below.

[0509] [ka]

[0510] (Ab comprises an antibody or its antigen-binding fragment, M ’is a linking group, and Sp is -L a -Q a - where A is selected from none, hydrogen, or alkyl, T represents a linear or branched (C1-C8) trivalent alkyl group, and Y, Y', and Y'' are -L respectively. b -Q b - and L a , L b Each is selected from none, alkylene, alkenylene, alkylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, heteroarylene, or polyethylene glycol group, where the alkylene, alkenylene, alkylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, heteroarylene, and polyethylene glycol groups are optionally substituted with substituents, Q a Q b Each is selected from none, -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-, where R is selected from hydrogen or optionally substituted alkyl, and Z is none. Z', Z'' are selected from hydrogen, optionally substituted alkyl, or sulfonic acid internal salts, none, hydrogen, optionally substituted alkyl, sulfonic acid internal salt, or phosphate internal salt, at least one of Z, Z', Z'' is a sulfonic acid internal salt or phosphate internal salt, X contains a self-destructing unit or none, D contains a drug molecule, and i is an integer of 1 or more, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, etc.

[0511] h is an integer between 1 and 6, for example, 1, 2, 3, 4, 5, 6, etc. o, p, and q are each integers between 0 and 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.

[0512] If h is greater than 1, then multiple Sps may have different structures or the same structure, i.e., multiple Ls a These may be different structures or identical structures, and there may be multiple Qs. a The structures may be different or identical. a and Q a The number of each can be the same or different.

[0513] If i is greater than 1, then 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.

[0514] If p is greater than 1, multiple Y's may have different structures or the same structure.

[0515] If j is greater than 1, the parts in parentheses may have the same structure or different structures.

[0516] In this application, formula (2) is formula (2-1).

[0517] [ka]

[0518] (Here, Ab, M ’ The definitions of Sp, A, T, Y, h, o, i, j, X, D, and Z are the same as those given in equation (1) above. At least one of the i Zs is an intrasulfonic acid salt.

[0519] Furthermore, equation (2-1) is equivalent to equation (2-1-1).

[0520] [ka]

[0521] (Here, AA is an amino acid residue, w is an integer greater than or equal to 1, such as 1, 2, 3, 4, 5, or 6. If w is greater than 1, the multiple AAs may be residues of the same amino acid or different amino acids.

[0522] In this application, formula (2) is formula (2-2).

[0523] [ka]

[0524] (Here, Ab, M ’ The definitions of Sp, A, T, Y, Z, o, p, i, h, j, X, Y', and D are the same as those described in formula (1) above, where Z' is an end-sodium sulfonic acid or end-sodium phosphate.

[0525] Furthermore, equation (2-2) is equation (2-2-1).

[0526] [ka]

[0527] (Here, AA are amino acid residues, and w is an integer greater than or equal to 1, such as 1, 2, 3, 4, 5, 6, etc. If w is greater than 1, multiple AAs may be residues of the same amino acid or residues of different amino acids.)

[0528] In this application, formula (2) is formula (2-3).

[0529] [ka]

[0530] (Here, Ab, M ’The definitions of Sp, A, Y, T, Z, o, i, h, j, q, X, Y'', and D are the same as in the explanation of formula (1) above, where Z'' is an end-sulfonic acid salt or an end-phosphate salt.

[0531] Furthermore, equation (2-3) is equivalent to equation (2-3-1).

[0532] [ka]

[0533] (Here, AA are amino acid residues, and w is an integer greater than or equal to 1, such as 1, 2, 3, 4, 5, 6, etc. If w is greater than 1, multiple AAs may be residues of the same amino acid or residues of different amino acids.)

[0534] In this specification, all definitions given for formula (1) apply to formula (2), all definitions given for formula (1-1) apply to formula (2-1), all definitions given for formula (1-1-1) apply to formula (2-1-1), all definitions given for formula (1-2) apply to formula (2-2), all definitions given for formula (1-2-1) apply to formula (2-2-1), and all definitions given for formula (1-3) apply to formula (1-3). All the definitions stated apply to equation (2-3), and all the definitions explained with respect to equation (1-3-1) apply to equation (2-3-1). Therefore, for Sp, A, Y, T, Z, o, i, h, j, q, X, Y', Z', Y'', Z'', and D in equations (2), (2-1), (2-1-1), (2-2), (2-2-1), (2-3), and (2-3-1), we refer to the explanation in equation (1) and do not further limit them here.

[0535] In this application, M ’ It is one of the following structures.

[0536] [ka]

[0537] (* is concatenated to Ab.) Furthermore, M ’ It is one of the following structures.

[0538] [ka]

[0539] In some embodiments, M ’ teeth,

[0540] [ka]

[0541] That is the case. In this application, the compound is selected from the following structures.

[0542] [ka]

[0543] [ka]

[0544] [ka]

[0545] In this application, the antibody is selected from the group consisting of mouse antibodies, chimeric antibodies, humanized antibodies, and fully human antibodies. In this application, the antibody includes monoclonal antibodies. In this application, the antibody includes bispecific antibodies. In this application, the antigen-binding fragment is Fab, Fab ’ , Fv fragment, F(ab ’ The group is selected from )2, F(ab)2, scFv, di-scFv, and VHH.

[0546] In the present application, the Ab is selected from the group consisting of an anti-Claudin 18.2 antibody, an anti-human folate receptor antibody, an anti-HER2 antibody, an anti-HER3 antibody, an anti-TROP2 antibody, an anti-B7-H3 antibody, an anti-GPC20 antibody, an anti-CDH6 antibody, an anti-EGFR antibody, an anti-EGFRvIII antibody, an anti-AXL antibody, an anti-Nectin-4 antibody, an anti-Tissue factor antibody, an anti-TIM-1 antibody, an anti-PSMA antibody, an anti-EpCAM antibody, an anti-MUC1 antibody, an anti-STEAP1 antibody, an anti-GPNMB antibody, an anti-FGF2 antibody, an anti-FOLR1 antibody, an anti-c-MET antibody, an anti-GFR antibody, an anti-AGS-16, an anti-Guanylyl cyclase C antibody, an anti-Mesothelin antibody, an anti-SLC44A4 antibody, an anti-EphA2 antibody, an anti-AGS-5 antibody, an anti-GPC-3 antibody, an anti-c-KIT antibody, an anti-ROR1 antibody, an anti-PD-L1 antibody, an anti-CD27L antibody, an anti-5T4 antibody, an anti-Mucin 16 antibody, an anti-NaPi2b antibody, an anti-STEAP antibody, an anti-SLITRK6 antibody, an anti-ETBR antibody, an anti-BCMA antibody, an anti-CEACAM5 antibody, an anti-SC-16 antibody, an anti-SLC39A6 antibody, an anti-Delta-like protein3 antibody, an anti-Claudin18.2 antibody, an anti-CD19 antibody, an anti-CD20 antibody, an anti-CD22 antibody, an anti-CD30 antibody, an anti-CD33 antibody, an anti-CD37 antibody, an anti-CD45 antibody, an anti-CD56 antibody, an anti-CD66e antibody, an anti-CD70 antibody, an anti-CD73 antibody, an anti-CD74 antibody, an anti-CD79b antibody, an anti-CD138 antibody, an anti-CD147 antibody, an anti-CD166 antibody, an anti-CD223 antibody, an anti-MUC16 antibody, an anti-MSLN antibody, an anti-ENPP3 antibody, an anti-SLTRK6 antibody, an anti-FGFR antibody, an anti-LIV-1 antibody, an anti-Lewis Y antibody, an anti-av-integrin antibody, an anti-ASCT2 antibody, an anti-C4.4a antibody, an anti-CA-IX antibody, an anti-CD324 antibody, an anti-CD352 antibody, an anti-CD44v6 antibody, an anti-CD48a antibody, an anti-CLL-1 antibody, an anti-Cripto antibody, an anti-CS1 antibody, an anti-DPEP3 antibody, an anti-Ephrin-A2 antibody, an anti-Ephrin-A4 antibody, an anti-ETBR antibody, an anti-FGFR2 antibody, an anti-FGFR3 antibody, an anti-FLT3 antibody, an anti-GD3 antibody, an anti-GloboH antibody, an anti-GPC3 antibody, an anti-LAMP-1 antibody, an anti-LRRC15 antibody, an anti-Ly6E antibody, an anti-MFI2 antibody, an anti-NOTCH3 antibody, an anti-p-cadherin antibody, an anti-PRLR antibody, an anti-RNF43 antibody, and antigen-binding fragments of the above antibodies.

[0547] In this application, the heavy chains HCDR1, HCDR2, and HCDR3 of Ab, and the light chains LCDR1, LCDR2, and LCDR3 respectively include the heavy chains HCDR1, HCDR2, and HCDR3 and the light chains LCDR1, LCDR2, and LCDR3 of the antibody.

[0548] In this application, the heavy chain variable region VH and the light chain variable region VL of Ab each include the heavy chain variable region VH and the light chain variable region VL of the antibody, respectively. In this application, the heavy chain and the light chain of Ab each include the heavy chain and the light chain of the antibody, respectively. In this application, Ab is an anti-claudin 18.2 antibody or its antigen-binding fragment. In this application, Ab is an anti-human folate receptor antibody or its antigen-binding fragment.

[0549] The present invention also provides pharmaceutical compositions comprising the aforementioned compounds, or their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

[0550] The present invention also provides the use of the aforementioned compounds, or in the form of their tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates, in the preparation of drugs for treating and / or preventing tumors.

[0551] In this application, the tumor is defined as containing 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 The target is selected from tumors associated with the expression of a group of targets consisting of H, GPC3, LAMP-1, LRRC15, Ly6E, MFI2, NOTCH3, p-cadherin, PRLR, and RNF43.

[0552] In this application, tumors associated with the expression of the target include tumors in which the target is highly expressed and / or tumors in which the target is positive.

[0553] In this application, the tumor is selected from solid tumors, hematological malignancies, and metastatic, refractory, or recurrent lesions of cancer.

[0554] In this application, the tumor is selected from the group consisting of esophageal cancer, digestive tract cancer, pancreatic cancer, thyroid cancer, colorectal cancer, kidney cancer, lung cancer (e.g., non-small cell lung cancer), liver cancer, stomach 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. [Examples]

[0555] Examples Unless otherwise specified, the experimental methods used in the following examples are all standard procedures.

[0556] Unless otherwise specified, all materials, reagents, etc., used in the following examples are commercially available.

[0557] Fmoc-L-glutamic acid-5-tert-butyl ester, compound 12-1, HATU (Japanese name: O-(7-azabenzotriazole-1-yl)-N,N,N) in the following examples. ’ ,N ’-Tetramethyluronium hexafluorophosphate), DIEA (Japanese name: N,N-diisopropylethylamine), di(p-nitrophenyl) carbonate, exatecan mesylate, Mal-amido-PEG4-acid (Japanese name: maleimido-amide-PEG4-propionic acid), DCM (Japanese name: dichloromethane), TFA (Japanese name: 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 (Japanese name: Monomethyl auristatin E), HOBT (Japanese name: 1-hydroxybenzotriazole), methyl tert-butyl ether, compound 65-1, BH3-Me2S, DMAP (Japanese name: 4-dimethylaminopyridine), (BOC)2O, EA (Japanese name: Ethyl acrylate), TBAF (Japanese name: Tetra-n-butylammonium Compound 71-1, 6-maleimidocaproic acid N-succinimidyl ester, Compound 72-1, HOSu (Japanese name: N-hydroxysuccinimid), EDCI (Japanese name: carbodiimide), Fmoc-glycine, nitrophenyl chloroformate, Compound 73-1, BCN-PEG4-acid (Japanese name: rel-1-((1R,8S,9s)-bicyclo[6 [1.0] Non-4-in-9-yl)-3-oxo-2,7,10,13,16-pentaoxa-4-azanonadecane-19-acid), Fmoc-NMe-PEG4-C2-Acid, N,N-diisopropylethylamine, 4-methylsulfonylbenzoic acid, DIPEA (Japanese name: N,N-diisopropylethylamine), exo-3,6-epoxy-1,2,3,6-tetrahydrophthalic anhydride, and N-Boc-N-methylethylenediamine are all commercially available.

[0558] Example 1 (Preparation of LD-12) The synthesis scheme for LD-12 is as follows:

[0559] [ka]

[0560] 12-2 synthesis Fmoc-L-glutamic acid-5-tert-butyl ester (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, and the mixture was stirred continuously at room temperature for 18 hours. LC-MS confirmed that the reaction had proceeded completely. After adding water and extracting three times with ethyl acetate, the organic phases were washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 0.9 g of crude product.

[0561] 12-3 synthesis Compound 12-2 (300 mg, 0.43 mmol), di(p-nitrophenyl) 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. LC-MS confirmed that the reaction had proceeded completely. After adding water and extracting three times with ethyl acetate, the organic phases were washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification by thin-layer TLC plate (MeOH / DCM, 0%~3%) yielded 0.3 g. MS+Na(+)889.4.

[0562] 12-4 synthesis Compound 12-3 (350 mg, 0.425 mmol), DIEA (207 mg, 1.61 mmol), and exatecan mesylate (600 mg, 0.69 mmol) were dissolved in 2 mL of anhydrous DMF and reacted at room temperature for 6 hours. After adding water, the mixture was extracted three times with ethyl acetate. The organic phases were washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The product was purified by normal-phase silica gel column (MeOH / DCM, 0%~10%) to obtain 470 mg. MS (+) 1162.4.

[0563] 12-5 synthesis Compound 12-4 (100 mg, 0.086 mmol) was dissolved in 2 mL of anhydrous DMF, and piperidine (0.05 mL) was added at room temperature and the mixture was stirred for 2 hours. After concentration under reduced pressure, the mixture was slurryed with methyl tert-butyl ether and filtered to obtain 80 mg of crude product, which was used directly in the next step of the reaction. MS (+) 940.4.

[0564] 12-6 synthesis 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 5 minutes. Mal-amido-PEG4-acid (80 mg, 0.085 mmol) and DIEA (33 mg, 0.26 mmol) were added sequentially, and the mixture was stirred continuously at room temperature for 2 hours. LC-MS confirmed that the reaction was complete. After adding water and extracting three times with ethyl acetate, the organic phases were washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by thin-layer TLC plate (MeOH / DCM, 0%~3%) to obtain 30 mg. MS (+) 1339.2.

[0565] Synthesis of LD-12 Compound 12-6 (30 mg, 0.022 mmol) was dissolved in 2 mL of DCM, and 0.3 mL of TFA was added at room temperature. The mixture was stirred for 2 hours, and LC-MS confirmed that the reaction had proceeded completely. After concentration under reduced pressure, 7.7 mg was obtained by reverse-phase preparative chromatography (column chromatography) (mobile phase containing 0.1% formic acid). LCMS:m / z(ES+)(M+H)+=1282.4, Rt=1.385 min 1H NMR(400 MHz, DMSO-d6)δ 10.00 (s, 1H), 8.23 ​​(d, J = 6.8 Hz, 1H), 8.11-8.03 (m, 3H), 7.79 (d, J = 10.8 Hz, 1H), 7.72 (d, J = 8.4 Hz, 1H), 7.60 (d, J = 8.4 Hz, 2H), 7.38 (d, J = 8.4 Hz, 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 = 7.2 Hz,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 = 6.8 Hz, 1H), 1.31 (d, J = 7.2 Hz, 3H), 0.91-0.83 (m, 9H).

[0566] Example 2 (Preparation of LD-27) The LD-27 process is as follows:

[0567] [ka]

[0568] 27-1 synthesis 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 the mixture was stirred continuously at room temperature for 3 hours. LC-MS confirmed that the reaction had proceeded completely. After adding water and extracting three times with ethyl acetate, the organic phases were washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 80 mg of crude product. MS(+) = 1133.4.

[0569] Synthesis of LD-27 Compound 27-1 (70 mg, 0.062 mmol) was dissolved in 5 mL of DCM, and 0.8 mL of TFA was added at room temperature. The mixture was stirred for 2 hours, and LC-MS confirmed that the reaction had proceeded completely. After concentration under reduced pressure, 22.8 mg was obtained by reverse-phase preparative fractionation (mobile phase containing 0.1% formic acid). LCMS:m / z(ES+)(M+H)+=1077.4, Rt=1.454 min 1H NMR(400 MHz, DMSO-d6)δ 12.08 (s, 1H), 9.97 (s, 1H), 8.21 (d, J = 6.8Hz, 1H), 8.06-7.98 (m, 2H), 7.77 (d, J = 10.8 Hz, 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 = 4.4 Hz, 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).

[0570] Example 3 (Preparation of LD-31) The synthesis scheme for LD-31 is as follows:

[0571] [ka]

[0572] 31-2 synthesis Fmoc-L-valine-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 ChemExpress Co., Ltd., see WO2022058548A1 for synthesis method) was added to the reaction mixture. After stirring at room temperature for 2 hours, the reaction was confirmed to be complete as monitored by LC-MS. A concentrated crude product was obtained. It was purified by normal-phase silica gel column (MeOH / DCM, 0%~10%) to obtain 320 mg. MS+Na(+)=638.4.

[0573] 31-3 synthesis Compound 31-2 (320 mg, 0.520 mmol) was dissolved in 5 mL of DCM, and di(p-nitrophenyl) carbonate (474.30 mg, 1.559 mmol) and DIEA (0.344 mL, 2.079 mmol) were added sequentially, and the mixture was stirred at room temperature for 2 hours. Monitoring by LC-MS confirmed that the reaction was complete. Saturated saline solution 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. The crude product was purified by normal-phase silica gel column (MeOH / DCM, 0%~10%) to obtain 340 mg. MS+Na(+) = 803.2.

[0574] 31-4 synthesis Compound 31-3 (340 mg, 0.462 mmol) was dissolved in 2 mL of anhydrous DMF, and exatecan 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 monitored by LC-MS and confirmed to be complete. Saturated saline solution was added to precipitate the solid crude product. The crude product was slurryed with methyl tert-butyl ether to obtain 157 mg of compound 31-4. MS(+) = 1078.2.

[0575] 31-5 synthesis 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 mixture. The mixture was stirred at room temperature for 1 hour. The mixture was concentrated to obtain 292 mg of crude product, which was used directly in the next step of the reaction. MS(+) = 855.2.

[0576] 31-6 synthesis 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, and the mixture was stirred at room temperature for 2 hours. The reaction was completely carried out by LC-MS. Saturated saline solution was added to precipitate the solid crude product. The crude product was slurryed with methyl tert-butyl ether to obtain 350 mg of the crude product. MS (+) = 1254.4.

[0577] 31-7 synthesis Compound 31-6 (90 mg crude product) was dissolved in 2.5 mL of anhydrous DCM, and TFA (0.5 mL) was added to the reaction mixture. The mixture was stirred at room temperature for 1 hour. The mixture was concentrated to obtain 80 mg of crude product, which was used directly in the next step of the reaction. MS(+) = 1198.4.

[0578] Synthesis of LD-31 Compound 31-7 (76.62 mg, 0.064 mmol) was dissolved in 2 mL of anhydrous DMF, and DIEA (16.54 mg, 0.128 mmol) and HATU (29.20 mg, 0.077 mmol) were added, and the mixture was stirred at room temperature for 15 minutes. Quaternary ammonium sulfonic acid salt A (14.81 mg, 0.070 mmol, CAS number: 78276-19-4, supplied by ShangHai HaoYuan ChemExpress, see WO2011146595A2 for synthesis method) was added, and the mixture was stirred at room temperature for 2 hours. LC-MS confirmed that the reaction had proceeded completely. After adding water and extracting three times with ethyl acetate, the organic phases were washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The solution was purified by reverse-phase preparative fractionation (formic acid) to obtain a white solid LD-31 (P1: 12.5 mg, P2: 11.3 mg). LCMS: m / z (ES+)(M+H)+ = 1390.6, Rt = 1.242 min (P1), 1.279 min (P2).

[0579] Example 4 (Preparation of LD-37) The synthesis scheme for LD-37 is as follows:

[0580] [ka]

[0581] Synthesis of LD-37 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, and DIEA (9.88 mg, 0.076 mmol) and HATU (17.43 mg, 0.046 mmol) were added, and the mixture was stirred at room temperature for 15 minutes. Quaternary ammonium sulfonic acid salt A (8.04 mg, 0.038 mmol, CAS number: 78276-19-4, provided by ShangHai HaoYuan ChemExpress; see WO2011146595A2 for the synthesis method) was added, and the mixture was stirred at room temperature for 2 hours. LC-MS confirmed that the reaction had proceeded completely. After adding water and extracting three times with ethyl acetate, the organic phases were washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The solution was purified by reverse-phase preparative fractionation (with a mobile phase containing 0.1% formic acid) to obtain a white solid LD-37 (7.35 mg, 13.04% yield). LCMS: m / z (ES+)(M+H)+ = 1475.6, Rt = 1.344 min 1H NMR(400 MHz, DMSO)δ 10.03 (s, 1H), 8.24 (d, J = 6.4 Hz, 1H), 8.16 - 8.07 (m, 3H), 8.03 (t, J = 6.0 Hz, 1H), 7.80 (d, J = 10.8Hz, 1H), 7.71 (d, J = 9.2 Hz, 1H), 7.62 (d, J = 8.0 Hz, 2H), 7.38 (d, J = 8.4 Hz, 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 = 6.8 Hz, 1H), 4.31 (s, 1H), 4.24 - 4.20 (m, 1H), 3.61 (t, J = 7.2 Hz, 5H), 3.46-3.51 (m, 16H), 3.16 (dd, J = 11.2, 5.6 Hz, 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 = 9.2 Hz, 2H), 1.79 - 1.69 (m, 1H), 1.32 (d, J = 7.2 Hz, 3H), 0.89 (d, J = 6.8 Hz, 6H), 0.84 (d, J = 6.8 Hz, 3H).

[0582] Example 5: (modulation of LD-38) LD-38 is a synthesized product.

[0583]

change

[0584] 38-1のSynthesis Fmoc-L-glutamic acid-5-tert-butyl ester (144.7 mg, 1.175 mmol) and 4-aminobenzyl alcohol (500 mg, 1.175 mmol) were dissolved in 3 mL of anhydrous DMF, and DIEA (303.7 mg, 2.350 mmol) and HATU (536.19 mg, 1.410 mmol) were added. The mixture was stirred at room temperature for 16 hours, and LC-MS confirmed that the reaction had proceeded completely. After adding water and extracting three times with ethyl acetate, the organic phases were washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by normal-phase silica gel column (EA / PE, 0%~45%) to obtain 530 mg. MS(+) = 475.2.

[0585] 38-2 synthesis Compound 38-1 (530 mg, 0.999 mmol) was dissolved in 3 mL of anhydrous DMF, and piperidine (0.2 mL) was added to the reaction mixture. The mixture was stirred at room temperature for 1 hour. The mixture was concentrated to obtain 600 mg of crude product, which was used directly in the next step of the reaction. MS(+) = 309.2.

[0586] 38-3 synthesis Compound 38-2 (600 mg crude product) and Fmoc-L-valine (500 mg, 1.175 mmol) were dissolved in 3 mL of anhydrous DMF, and DIEA (334.13 mg, 2.585 mmol) and HATU (589.82 mg, 1.551 mmol) were added. The mixture was stirred at room temperature for 16 hours, and LC-MS confirmed that the reaction had proceeded completely. After adding water and extracting three times with ethyl acetate, the organic phases were washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. It was purified by normal-phase silica gel column (EA / PE, 40%~60%) to obtain 450 mg. 6. MS(+) = 630.4.

[0587] 38-4 synthesis Compound 38-3 (450 mg, 0.715 mmol) was dissolved in 1 mL of anhydrous DCM and 4 mL of anhydrous DMF, and di(p-nitrophenyl) carbonate (869.5 mg, 2.858 mmol) and 0.354 mL of DIEA were added sequentially. After reacting at room temperature for 12 hours, water was added to the reaction mixture, and the mixture was extracted three times with ethyl acetate. The organic phases were then washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by normal-phase silica gel column (EA / PE, 30%~70%) to obtain 200 mg. MS(+) = 543.2.

[0588] 38-5 synthesis Compound 38-4 (100 mg, 0.126 mmol) and exatecan mesylate (54.78 mg, 0.126 mmol) were dissolved in 2 mL of anhydrous DMF, and 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, water was added to the reaction mixture, and the mixture was extracted three times with ethyl acetate. The organic phases were then washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by normal-phase silica gel column (EA / PE, 30%~70%) to obtain 100 mg. MS(+) = 1091.4.

[0589] 38-6 synthesis Compound 38-5 (100 mg, 0.073 mmol) was dissolved in 3 mL of anhydrous DMF, and piperidine (0.3 mL) was added to the reaction mixture. The mixture was stirred at room temperature for 1 hour. The mixture was concentrated to obtain 110 mg of crude product, which was used directly in the next step of the reaction. MS(+) = 870.8.

[0590] 38-7 synthesis Compound 38-6 (110 mg crude product) and Mal-amido-PEG4-acid (30.19 mg, 0.073 mmol) were dissolved in 2 mL of anhydrous DMF, and DIEA (18.74 mg, 0.145 mmol) and HATU (33.08 mg, 0.087 mmol) were added. The mixture was stirred at room temperature for 2 hours, and LC-MS confirmed that the reaction had proceeded completely. After adding water and extracting three times with ethyl acetate, the organic phases were washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The product was purified by normal-phase silica gel column (MeOH / DCM, 5%~20%) to obtain 90 mg. MS(+) = 1267.6.

[0591] 38-8 synthesis 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. The mixture was concentrated to obtain 80 mg of crude product, which was used directly in the next step of the reaction. MS(+) = 2111.4.

[0592] Synthesis of LD-38 Compound 38-8 (80 mg, crude product) and quaternary ammonium sulfonic acid salt A (13.89 mg, 0.066 mmol, CAS number: 78276-19-4, supplied by ShangHai HaoYuan ChemExpress, see WO2011146595A2 for synthesis method) were dissolved in 2 mL of anhydrous DMF, and 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, and LC-MS confirmed that the reaction had proceeded completely. The mixture was purified by reverse-phase fractionation (mobile phase containing 0.1% formic acid) to obtain 13.5 mg of a white solid. LCMS: m / z (ES+)(M+H)+ = 1404.6, Rt = 1.314 min 1H NMR(400 MHz, DMSO)δ 9.89 (s, 1H), 8.18 (d, J = 7.2 Hz, 2H), 8.10 - 7.93 (m, 3H), 7.78 (d, J = 10.8 Hz, 1H), 7.63 (d, J = 8.4 Hz, 2H), 7.37 (d, J = 8.4 Hz, 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, 7.6 Hz, 1H), 4.20 - 4.14 (m, 1H), 3.59 (dd, J = 14.8, 7.6 Hz, 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).

[0593] Example 6 (Preparation of LD-57) The synthesis scheme for LD-57 is as follows:

[0594] [ka]

[0595] 57-1 synthesis Compound 38-4 (288 mg, 0.363 mmol; see Example 5 for its synthesis method) and MMAE (200 mg, 0.279 mmol) were dissolved in 5 mL of anhydrous DMF, and pyridine (1 mL) and HOBT (59 mg, 0.279 mmol) were added sequentially. The mixture was reacted at room temperature for 18 hours to obtain a concentrated crude product. It was purified by normal-phase silica gel column (MeOH / DCM, 0%~10%) to obtain 320 mg. MS(+) = 1374.2.

[0596] 57-2 synthesis Compound 57-1 (260 mg, 0.189 mmol) was dissolved in 3 mL of anhydrous DMF, and piperidine (0.05 mL) was added to the reaction mixture. The mixture was stirred at room temperature for 1 hour. The mixture was concentrated and slurryed with methyl tert-butyl ether to obtain 215 mg of crude product, which was used directly in the next step of the reaction. MS(+) = 1152.5.

[0597] 57-3 synthesis 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, and DIEA (0.092 mL, 0.559 mmol) and HATU (142 mg, 0.374 mmol) were added. The mixture was stirred at room temperature for 2 hours, and LC-MS confirmed that the reaction had proceeded completely. After adding water and extracting three times with ethyl acetate, the organic phases were washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The product was purified by normal-phase silica gel column (MeOH / DCM, 0%~10%) to obtain 170 mg. MS(+) = 1345.7.

[0598] 57-4 synthesis Compound 57-3 (150 mg, 0.112 mmol) was dissolved in 3 mL of anhydrous DCM, and TFA (0.6 mL) was added to the reaction mixture. The mixture was 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 step of the reaction. MS(+) = 1289.5.

[0599] Synthesis of LD-57 Compound 57-4 (143 mg, 0.111 mmol) and quaternary ammonium sulfonic acid salt A (70 mg, 0.333 mmol, CAS number: 78276-19-4, supplied by ShangHai HaoYuan ChemExpress; see WO2011146595A2 for synthesis method) were dissolved in 2 mL of anhydrous DMF, and 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, and LC-MS confirmed that the reaction had proceeded completely. The mixture was purified by reverse-phase fractionation (mobile phase containing 0.1% formic acid) to obtain 30 mg of a white solid. LCMS:m / z(ES+)(M+H)+=1481.2, Rt=1.475 min 1H NMR(400 MHz, DMSO)δ 9.89 (s, 1H), 8.40 - 8.03 (m, 3H), 7.92 (t, J = 7.6 Hz, 1H), 7.63 (d, J = 6.4 Hz, 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 = 14.4 Hz, 3H), 2.37 - 1.68 (m, 17H), 1.61 - 1.45 (m, 6H), 1.40 - 0.70 (m, 36H).

[0600] Example 7 (Preparation of LD-65) The synthesis scheme for LD-67 is as follows:

[0601] [ka]

[0602] 65-2 synthesis 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 mixture was filtered to obtain the crude product. It was washed twice with water and dried under reduced pressure to obtain 550 mg. MS(+) 211.2.

[0603] 65-3 synthesis Compound 65-2 (500 mg, 2.379 mmol) was dissolved in 50 mL of anhydrous THF, and BH3-Me2S (457 mg, 5.947 mmol) (10.0 M in THF) was slowly added dropwise under ice bath. After refluxing for 5 hours, LC-MS confirmed that the reaction had proceeded completely. A 4 M HCl (12 mL) methanol solution was added to the reaction mixture under ice bath, and the reaction was continued at 65°C for 8 hours. The mixture was filtered and dried under reduced pressure to obtain 350 mg. MS(+) 197.08.

[0604] 65-4 synthesis Compound 65-3 (350 mg crude product) and imidazole (485.77 mg, 7.135 mmol) were dissolved in 5 mL of anhydrous DCM, and TBDPS-Cl (735.44 mg, 2.676 mmol) was added at 0°C. The mixture was stirred for 2 hours, and LC-MS confirmed that the reaction had proceeded completely. After adding water and extracting three times with ethyl acetate, the organic phases were washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The product was purified by normal-phase silica gel column (EA / PE, 10%~50%) to obtain 710 mg. MS(+) 435.20.

[0605] 65-5 synthesis Compound 64-4 (1.5 g, 3.451 mmol) and DMAP (0.21 g, 1.726 mmol) were dissolved in DCM (20 mL), and (BOC)2O (1.51 g, 6.903 mmol) was added and the mixture was stirred for 2 hours. After adding water and extracting three times with ethyl acetate, the organic phases were washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by normal-phase silica gel column (EA / PE, 10%~50%) to obtain 1.87 g. MS+Na(+)557.3.

[0606] 65-6 synthesis 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 in an evaporator to obtain 1.7 g. MS+Na(+)527.6.

[0607] 65-7 synthesis 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. The mixture was stirred at room temperature for 4 hours. The reaction mixture was dried using an evaporator and purified by normal-phase silica gel column chromatography (MeOH / DCM, 0%~10%) to obtain 170 mg. MS+Na(+) = 289.2.

[0608] 65-8 synthesis 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 the mixture was stirred at room temperature for 16 hours. The reaction mixture was dried using an evaporator and purified by normal-phase silica gel column chromatography (MeOH / DCM, 0%~10%) to obtain 367 mg. MS(+)Na(+) = 681.4.

[0609] 65-9 synthesis Compound 65-9 (367 mg, 0.557 mmol), di(p-nitrophenyl) 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. LC-MS confirmed that the reaction had proceeded completely. After adding water and extracting three times with ethyl acetate, the organic phases were washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification by normal-phase silica gel column (MeOH / DCM, 0%~10%) yielded 380 mg. MS(+)Na(+) = 846.4.

[0610] 65-10 synthesis Compound 65-9 (350 mg, 0.425 mmol), DMAP (5.19 mg, 0.043 mmol), and exatecan mesylate (248.50 mg, 0.468 mmol) were dissolved in 2 mL of anhydrous DMF and reacted at room temperature for 2 hours. After adding water, the mixture was extracted three times with ethyl acetate. The organic phases were washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The product was purified by normal-phase silica gel column (MeOH / DCM, 0%~10%) to obtain 350 mg. MS(+) 1120.6.

[0611] 65-11 synthesis Compound 65-10 (350 mg, 0.312 mmol) was dissolved in 2 mL of anhydrous DMF, pyridine (0.05 mL) was added at room temperature, and the mixture was stirred for 2 hours. After concentration under reduced pressure, 380 mg of the crude product was used directly in the next reaction. MS(+) = 898.41.

[0612] 65-12 synthesis 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 5 minutes. Mal-amido-PEG4-acid (250 mg, 0.278 mmol) and DIEA (3 mL) were added sequentially, and the mixture was stirred continuously at room temperature for 2 hours. LC-MS confirmed that the reaction had proceeded completely. After adding water and extracting three times with ethyl acetate, the organic phases were washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by normal-phase silica gel column (MeOH / DCM, 0%~10%) to obtain 185 mg. MS (+) = 1296.6.

[0613] 65-13 synthesis 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. LC-MS confirmed that the reaction was complete. The solution was concentrated under reduced pressure to obtain 37 mg. MS+Na(+) = 1218.6.

[0614] Synthesis of LD-65 Quaternary ammonium sulfonic acid salt B (37 mg, 0.031 mmol, CAS number: 1349808-80-5, supplied by ShangHai HaoYuan ChemExpress; see WO2011146595A2 for synthesis method) was dissolved in 2 mL of anhydrous DMF, and HATU (12.94 mg, 0.034 mmol) was added, and the mixture was stirred at room temperature for 5 minutes. Compound 65-13 (11.75 mg, 0.046 mmol) and DIEA (0.008 mL, 0.046 mmol) were added sequentially, and the mixture was stirred continuously at room temperature for 2 hours. LC-MS confirmed that the reaction was complete. Reverse phase fractionation (mobile phase containing 10 mM NH4OAc) yielded 5.2 mg of a white solid. LCMS: m / z (ES+)(M+H)+ = 1432.6, Rt = 1.285 min 1H NMR(400 MHz, DMSO)δ 10.2-9.9 (m,1H), 8.19 (t, J = 7.2 Hz, 1H), 8.12 - 8.00 (m, 2H), 7.90 - 7.85 (m, 1H), 7.78 (d, J = 10.8 Hz, 1H), 7.39 - 7.30 (m, 3H), 7.00 (s, 2H), 6.52 (s, 1H), 5.44 (d, J = 4.0 Hz, 2H), 5.29 (s, 4H), 5.12 (d, J = 10.8 Hz, 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).

[0615] Example 8 (Preparation of LD-66) The synthesis scheme for LD-66 is as follows:

[0616] [ka]

[0617] Synthesis of LD-66 Quaternary ammonium phosphate (8.93 mg, 0.046 mmol, CAS number: 853798-56-8, supplied by ShangHai HaoYuan ChemExpress; see WO2005114193 for synthesis method) was dissolved in 2 mL of anhydrous DMF, and DIEA (0.008 mL, 0.046 mmol) and HATU (12.94 mg, 0.034 mmol) were added, and the mixture was stirred at room temperature for 5 minutes. Compound 65-13 (37 mg, 0.031 mmol) was added, and the mixture was stirred for 2 hours. LC-MS confirmed that the reaction was complete. The mixture was purified by reverse-phase preparative fractionation (formic acid) to obtain 11.5 mg of a white solid. LCMS: m / z (ES+)(M+H)+ = 1420.6, Rt = 1.246 min 1H NMR(400 MHz, DMSO)δ 10.11 (s, 1H), 8.23-8.17 (m,1H), 8.07 - 7.87 (m, 2H), 7.79 - 7.69 (m, 2H), 7.45 - 7.13 (m, 4H), 7.00 (s, 2H), 6.51 (s, 1H), 5.47 - 5.06 (m, 8H), 4.83 (dd, J = 45.6, 17.8 Hz, 1H), 4.59 (d, J = 7.6 Hz, 1H), 4.48 (d, J = 8.4 Hz, 1H), 4.37 (d, J = 6.8 Hz, 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 = 6.8 Hz, 3H), 0.90-0.82 (m, 9H).

[0618] Example 9 (Preparation of LD-71) The synthesis scheme for LD-71 is as follows:

[0619] [ka]

[0620] 71-2 synthesis Compound 31-1 (240 mg, 1.075 mmol; see Example 1 for its synthesis) and Compound 71-1 (267 mg, 0.538 mmol; provided by ShangHai HaoYuan ChemExpress; see WO2022058548A1 for its synthesis) 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 to precipitate a solid, and the reaction was confirmed to be complete by monitoring with LC-MS. The mixture was filtered, the filtered cake was washed three times with DCM, and dried under reduced pressure to obtain 260 mg. MS(+) = 702.4.

[0621] 71-3 synthesis Compound 71-2 (240 mg, 0.342 mmol), di(p-nitrophenyl) 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 by LC-MS and confirmed to be complete. After adding water and extracting three times with ethyl acetate, the organic phases were washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The product was purified by normal-phase silica gel column (MeOH / DCM, 5%~10%) to obtain 220 mg. MS(+) = 867.4.

[0622] 71-5 synthesis Under nitrogen protection at room temperature, triphosgene (136.1 mg, 0.459 mmol) was dissolved in 4 mL of anhydrous DCM, and 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 carried out at 50°C for 16 hours and monitored by LC-MS, confirming that the reaction was complete. After concentration and drying in an evaporator, the mixture was slurryed with methyl tert-butyl ether and filtered to obtain 127 mg. MS(+) = 607.4.

[0623] 71-6 synthesis Compound 71-5 (140 mg, 0.231 mmol) was dissolved in 3 mL of anhydrous DCM, and 0.6 mL of TFA was added at room temperature. The mixture was stirred for 2 hours, and LC-MS confirmed that the reaction had proceeded completely. The solution was concentrated under reduced pressure to obtain 116.9 mg. MS(+) = 507.2.

[0624] 71-7 synthesis 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, and HOBT (31.17 mg, 0.231 mmol) and DIEA (59.64 mg, 0.461 mmol) were added sequentially. After heating at 40°C and stirring for 3 hours, saturated saline solution was added to the reaction mixture to obtain the crude product. The product was purified by normal-phase silica gel column chromatography (MeOH, 0%~10%) to obtain 200 mg. MS(+) = 1234.6.

[0625] 71-8 synthesis Crude compound 71-7, 100 mg, was dissolved in 2 mL of anhydrous DMF, and pyridine (0.05 mL) was added at room temperature. The mixture was stirred for 2 hours. After concentration under reduced pressure, it was slurryed with methyl tert-butyl ether and filtered to obtain 100 mg of crude product, which was used directly in the next step. MS (+) 10¹².6.

[0626] 71-9 synthesis Crude compound 71-8 (100 mg) and 6-maleimidocaproate N-succinimidyl ester (43.9 mg, 0.142 mmol) were dissolved in 2 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 after extraction three times with ethyl acetate, the organic phases were washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by thin-layer silica gel plate (MeOH / DCM, 0%~10%) to obtain 60 mg. MS(+) = 1205.6.

[0627] 71-10 synthesis 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 the mixture was stirred for 2 hours. LC-MS confirmed that the reaction was complete. The solution was concentrated under reduced pressure to obtain 57 mg. MS(+) = 1149.6.

[0628] Synthesis of LD-71 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 5 minutes. Quaternary ammonium sulfonic acid salt A (15.64 mg, 0.074 mmol, CAS number: 78276-19-4, supplied by ShangHai HaoYuan ChemExpress, see WO2011146595A2 for synthesis method) and DIEA (0.025 mL, 0.149 mmol) were added sequentially, and the mixture was stirred continuously at room temperature for 2 hours. LC-MS confirmed that the reaction was complete. Reverse phase fractionation (mobile phase containing 0.1% formic acid) yielded 24.0 mg of a white solid. LCMS: m / z(ES+)(M+H)+=1341.6 ,Rt=1.210 min 1H NMR(400 MHz, DMSO)δ 10.04-9.96 (m, 1H), 8.45 (s, 1H), 8.185-8.085 (m, 2H), 7.968-7.782 (m, 3H), 7.63 - 7.32 (m, 9H), 6.99 (s, 2H), 6.53 (s, 1H), 5.99 (s, 1H), 5.70 (d, J = 20.6 Hz, 1H), 5.43-5.41 (m, 5H), 5.35 (s, 3H), 4.35-4.25 (m, 1H), 4.19-4.17 (m, 1H), 3.07-2.91 (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).

[0629] Example 10 (Preparation of LD-72) The synthesis scheme for LD-72 is as follows:

[0630] [ka]

[0631] 72-2 synthesis 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 1 hour, and then distilled under reduced pressure to form a slurry with petroleum ether to obtain 350 mg of crude product. MS+Na(+) = 357.9.

[0632] 72-3 synthesis 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, and DIEA (0.532 mL, 3.219 mmol) and HATU (817 mg, 2.150 mmol) were added. The mixture was stirred at room temperature for 2 hours, and LC-MS confirmed that the reaction had proceeded completely. After adding water and extracting three times with ethyl acetate, the organic phases were washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by normal-phase silica gel column (MeOH / DCM, 0%~5%) to obtain 400 mg. MS(+) = 529.6.

[0633] 72-4 synthesis Compound 72-3 (90 mg, 0.071 mmol) was dissolved in 3 mL of anhydrous DCM, and TFA (0.8 mL) was added to the reaction mixture. The mixture was 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 step of the reaction. MS(+) = 473.5.

[0634] 72-5 synthesis Compound 72-4 (150 mg, 0.317 mmol) was dissolved in 2 mL of HOSu (40.2 mg, 0.349 mmol) and EDCI (67.2 mg, 0.351 mmol), stirred at room temperature for 2 hours, and then distilled under reduced pressure to obtain 160 mg of crude product, which was used directly in the next step of the reaction. MS(+) = 569.8.

[0635] 72-6 synthesis 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 the mixture was stirred at room temperature for 18 hours. LC-MS confirmed that the reaction had proceeded completely. After adding water and extracting three times with ethyl acetate, the organic phases were washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by normal-phase silica gel column (MeOH / DCM, 0%~5%) to obtain 1000 mg. MS+Na(+) = 567.8.

[0636] 72-7 synthesis 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, water was added to the reaction mixture, and the mixture was extracted three times with ethyl acetate. The organic phases were then washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by normal-phase silica gel column (MeOH / DCM, 0%~5%) to obtain 300 mg. MS+Na(+) = 732.7.

[0637] 72-8 synthesis Compound 72-7 (340 mg, 0.478 mmol) and exatecan mesylate (254 mg, 0.478 mmol) were dissolved in 8 mL of anhydrous DMF, and 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, water was added to the reaction mixture, and the mixture was extracted three times with ethyl acetate. The organic phases were then washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The product was purified by normal-phase silica gel column (MeOH / DCM, 0%~10%) to obtain 380 mg. MS(+) = 1006.7.

[0638] 72-9 synthesis 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 1 hour, and then distilled under reduced pressure to form a slurry with petroleum ether to obtain 230 mg of crude product. MS(+) = 785.8.

[0639] 72-10 synthesis 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 after extraction three times with ethyl acetate, the organic phases were washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by thin-layer silica gel plate (MeOH / DCM, 0%~10%) to obtain 100 mg. MS(+) = 1240.8.

[0640] 72-11 synthesis Compound 71-10 (90 mg, 0.071 mmol) was dissolved in 3 mL of anhydrous DCM, and TFA (0.3 mL) was added to the reaction mixture. The mixture was 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 step of the reaction. MS(+) = 1138.6.

[0641] Synthesis of LD-72 Compound 71-11 (36 mg, crude product) and quaternary ammonium phosphate (11.4 mg, 0.047 mmol, Cas number: 853798-56-8, supplied by ShangHai HaoYuan ChemExpress; see WO2005114193 for synthesis method) were dissolved in 3 mL of anhydrous DMF, and 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, and LC-MS confirmed that the reaction had proceeded completely. The mixture was purified by reverse-phase fractionation (mobile phase containing 10 mM NH4OAc) to obtain 15 mg of a white solid. LCMS:m / z(ES+)(M+H)+=1361.5, Rt=1.114 min 1H NMR(400 MHz, DMSO)δ 10.01 (d, J = 12 Hz, 1H), 9.04 (d, J = 8.0 Hz, 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 = 13.6 Hz, 7H), 2.94 (d, J = 13.2 Hz, 9H), 2.40 - 2.33 (m, 3H), 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 = 7.2 Hz, 3H).

[0642] Example 11 (Preparation of LD-73) The synthesis scheme for LD-73 is as follows:

[0643] [ka]

[0644] Synthesis of LD-73 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 2 hours, monitoring by LC-MS confirmed that the reaction had proceeded completely, and the reaction system was subjected to reverse-phase fractionation (mobile phase containing 0.1% formic acid) to obtain 19 mg of a white solid. LCMS: m / z (ES+)(M+H)+ = 1634.8, Rt = 1.392 min 1H NMR(400 MHz, DMSO)δ 9.96 (d, J = 40.0 Hz, 1H), 8.17 (d, J = 6.8 Hz, 1H), 8.00 (s, 1H), 7.78 (d, J = 10.8 Hz, 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 - 3.55 (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).

[0645] Example 12 (Preparation of LD-74) The synthesis scheme for LD-74 is as follows:

[0646] [ka]

[0647] 74-1 synthesis 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 5 minutes. Compound 65-11 (72.11 mg, 0.080 mmol) and DIEA (0.020 mL) were added sequentially, and the mixture was stirred continuously at room temperature for 2 hours. LC-MS confirmed that the reaction had proceeded completely. After adding water and extracting three times with ethyl acetate, the organic phases were washed together with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by normal-phase silica gel column (MeOH / DCM, 0%~10%) to obtain 80 mg. MS (+) 1091.6.

[0648] 74-2 synthesis 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 the mixture was stirred for 2 hours. LC-MS confirmed that the reaction was complete. The solution was concentrated under reduced pressure to obtain 65 mg. MS (+) = 991.4.

[0649] Synthesis of LD-74 Compound 74-3 (20.00 mg, 0.026 mmol, purchased from ShangHai HaoYuan ChemExpress; see WO2022228493 for synthesis method) was dissolved in 1 mL of anhydrous DMF, and HATU (10.85 mg, 0.029 mmol) was added, and the mixture was stirred at room temperature for 5 minutes. Compound 74-2 (25.71 mg, 0.026 mmol) and DIEA (0.008 mL, 0.046 mmol) were added sequentially, and the mixture was stirred continuously at room temperature for 2 hours. LC-MS confirmed that the reaction had proceeded completely. The reaction mixture was subjected to reverse-phase fractionation (mobile phase containing 10 mM NH4OAc) to obtain 9.5 mg of a white solid. LCMS: m / z (ES+)(M+H)+ = 1743.9, Rt = 1.325 min 1H NMR(400 MHz, DMSO)δ 7.79 (d, J = 10.9 Hz, 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).

[0650] Example 13 (Preparation of LD-88) The synthesis scheme for LD-88 is as follows:

[0651] [ka]

[0652] Synthesis of LD-88 Compound 38-8 (50 mg, 0.041 mmol; see Example 3 for its synthesis method) and quaternary ammonium sulfonic acid salt C (16.34 mg, 0.078 mmol, CN115521998, ShangHai HaoYuan ChemExpress) were dissolved in 2 mL of anhydrous DMF, and 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, and LC-MS confirmed that the reaction had proceeded completely. The reaction mixture was purified by reverse-phase preparation (trifluoroacetic acid) to obtain a pale yellow solid LD-88 (10.0 mg, 17.28% yield). LCMS: m / z (ES+)(M+H)+ = 1402.6, Rt = 1.303 min 1H NMR(400 MHz, DMSO)δ 10.06 (s, 1H), 9.95 (s, 1H), 8.19 (d, J = 7.6 Hz, 1H), 8.10-8.00 (m, 2H), 7.91 (d, J = 8 Hz, 1H), 7.79 (d, J = 10.8 Hz, 1H), 7.61 (d, J = 8.4 Hz, 2H), 7.38 (d, J = 8.4 Hz, 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, 4.4 Hz, 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).

[0653] Example 14 (Preparation of LD-89) The synthesis scheme for LD-89 is as follows:

[0654] [ka]

[0655] Synthesis of LD-89 Compound 38-8 (50 mg, 0.041 mmol; see Example 3 for its synthesis method) and quaternary ammonium sulfonic acid salt D (17.58 mg, 0.078 mmol, Bioconjugate Chem. 2022, 33, 718-725, ShangHai HaoYuan ChemExpress) were dissolved in 2 mL of anhydrous DMF, and 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, and LC-MS confirmed that the reaction had proceeded completely. The reaction mixture was purified by reverse-phase preparative (formic acid) to obtain a white solid LD-89 (5.4 mg, 10.06% yield). LCMS: m / z (ES+)(M+H)+ = 1418.6, Rt = 1.288 min 1H NMR(400 MHz, DMSO)δ 9.86 (s, 1H), 8.11 (d, J = 7.2 Hz, 1H), 8.01 - 7.89 (m, 3H), 7.71 (d, J = 10.8 Hz, 1H), 7.55 (d, J = 8.4 Hz, 2H), 7.30 (d, J = 8.4 Hz, 2H), 7.24 (s, 1H), 6.93 (s, 2H), 6.45 (s, 1H), 5.38 (s, 2H), 5.22 (d, J = 4.0 Hz, 3H), 5.01 (s, 2H), 4.35 - 4.30 (m, 1H), 4.15-4.12 (m, 1H), 3.52 (s, 8H), 3.42 (d, J = 3.6 Hz, 17H), 3.25 (s, 5H), 3.09 - 2.99 (m, 14H), 2.31 (s, 4H), 2.26 (d, J = 7.2 Hz, 3H), 2.16-2.07 (m, 2H), 1.88-1.82 (m, 8H), 0.82-0.76 (m, 9H).

[0656] Example 15 (Preparation of LD-94) The synthesis scheme for LD-94 is as follows:

[0657] [ka]

[0658] Synthesis of compound 94-1 Compound 38-5 (450 mg, 0.412 mmol; see Example 3 for its synthesis method) was added to dichloromethane (5 mL), trifluoroacetic acid (1 mL) was added dropwise to the system, the mixture was stirred at room temperature for 2 hours, the system was concentrated and dried by aspirate, and the crude product was used in the next step. m / z (ES+)(M+1) = 1035.4

[0659] Synthesis of compound 94-2 Compound 94-1 (400 mg, 0.386 mmol), quaternary ammonium sulfonic acid salt 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, then 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 chromatography (water:acetonitrile 0%-70%) to obtain 170 mg. m / z (ES+)(M+1) = 1227.6.

[0660] Synthesis of compound 94-3 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 to the system. The mixture was stirred at room temperature for 2 hours, then concentrated and dried by suction. The mixture was then slurryed with petroleum ether, filtered, and the filtered cake was dried and used directly in the next step. m / z (ES+)(M+1) = 1005.6.

[0661] Synthesis of LD-94 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), stirred at room temperature for 5 minutes, then HATU (18.73 mg, 0.049 mmol) was added, and the mixture was reacted at room temperature for 2 hours. The product was then subjected to Prep-HPLC (NH4HCO3 system) to obtain LD-94 (6 mg, yield: 7.68%). LCMS: m / z (ES+)(M+H) + = 1427.6, Rt = 1.721 min. 1 H NMR(400 MHz, DMSO)δ 9.90 (s, 1H), 8.19 (s, 2H), 8.09 - 8.05 (m, 1H), 8.00 - 7.96 (m, 1H), 7.79 (d, J = 11.2 Hz, 1H), 7.63 (d, J = 7.6 Hz, 2H), 7.37 (d, J = 8.2 Hz, 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 = 8.6 Hz, 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).

[0662] Example 16 (Preparation of LD-98) The synthesis scheme for LD-98 is as follows:

[0663] [ka]

[0664] Synthesis of compound 98-1 Compound 65-11 (120 mg, 0.134 mmol; see Example 5 for its synthesis method) and Fmoc-NMe-PEG4-C2-Acid (CAS No.: 2170240-98-7, 67.0 mg, 0.134 mmol) were dissolved in DMF (2 mL), and 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. The mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM: MeOH, MeOH content 0-10%) to obtain 100 mg. m / z (ES+)(M+1) = 1381.6.

[0665] Synthesis of compound 98-2 Compound 98-1 (100 mg, 0.072 mmol) was dissolved in DMF (2 mL), and diethylamine (15.88 mg, 0.217 mmol) was added. The mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure to obtain 100 mg of crude product. m / z (ES+)(M+1) = 1159.6.

[0666] Synthesis of compound 98-3 Compound 98-2 (80 mg, 0.069 mmol) and 4-methylsulfonylbenzoic acid (13.95 mg, 0.069 mmol) were dissolved in DMF (2 mL), and HATU (28.86 mg, 0.076 mmol) and DIPEA (13.38 mg, 0.104 mmol) were added sequentially. 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 column (C18, 20-60% acetonitrile-H2O solution) to obtain 26 mg of a yellow solid. m / z (ES+)(M+1) = 1343.49.

[0667] Synthesis of compound 98-4 Compound 98-3 (25 mg, 0.019 mmol) was dissolved in DCM (2 mL), TFA (0.4 mL) was added, and the reaction mixture 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 step of the reaction. m / z (ES+)(M+1) = 1243.4.

[0668] Synthesis of LD-98 Compound 98-4 (23 mg, 0.018 mmol) and quaternary ammonium sulfonic acid salt B (7.03 mg, 0.028 mmol) were dissolved in DMF (1 mL), and 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 subjected to pre-HPLC (TFA) to obtain LD-98 (6 mg, yield: 22.2%). LCMS: m / z (ES+)(M+H) + = 1479.6, Rt = 1.340 min. 1H NMR(400 MHz, DMSO)δ 9.97 (d, J = 40.2 Hz, 1H), 9.14 (d, J = 12.0 Hz, 2H), 8.19 (t, J = 7.4 Hz, 1H), 8.06 (d, J = 9.4 Hz, 1H), 7.85 (s, 1H), 7.78 (d, J = 11.2 Hz, 1H), 7.64 (dd, J = 49.2, 8.4 Hz, 1H), 7.33 (d, J = 12.4 Hz, 2H), 6.52 (s, 1H), 5.44 (d, J = 4.0 Hz, 2H), 5.30 (s, 3H), 5.12 (d, J = 10.8 Hz, 2H), 4.64 (d, J = 26.4 Hz, 2H), 4.36 (dd, J = 13.6, 6.8 Hz, 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 = 9.2 Hz, 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, 1.8 Hz, 2H), 2.20 (s, 2H), 1.98 - 1.83 (m, 8H), 1.31 - 1.23 (m, 4H), 0.89 - 0.81 (m, 10H).

[0669] Example 17 (modulation of LD-99) LD-99 is a synthetic product.

[0670]

change

[0671] Synthesis of compound 99-1 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, and the reaction mixture was heated to 65 °C and reacted for 48 hours. After concentrating the reaction mixture, it was 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.

[0672] Synthesis of compound 99-2 Compound 99-1 (2.3 g, 7.135 mmol) was dissolved in dichloromethane (20 mL), and trifluoroacetic acid (4 mL) was added under ice water. The reaction mixture was then heated to room temperature and allowed to react for 1 hour. The reaction mixture was concentrated to obtain 1.5 g of the crude product. m / z(ES+)(M+H) + = 223.2.

[0673] Synthesis of compound 99-3 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. After concentrating the reaction mixture, it was purified by column chromatography (petroleum ether:ethyl acetate = 3:1) to obtain 2.5 g of the target product. m / z(ES+)(M+Na) + = 393.2.

[0674] Synthesis of compound 99-4 Compound 99-3 (1.5 g, 3.822 mmol) was dissolved in toluene (15 mL) and reacted at 110°C for 18 hours. The reaction mixture was concentrated to obtain 1 g of the target product as a yellow oily substance. m / z (ES+)(M+H) + = 325.2.

[0675] Synthesis of compound 99-5 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 mixture was concentrated to obtain 82 mg of the crude product as a yellow oily substance. m / z(ES+)(M+H) + = 269.2.

[0676] Synthesis of compound 99-6 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 sequentially. The reaction was allowed to proceed at room temperature for 2 hours. After filtering the reaction mixture, it was purified by reverse-phase column chromatography to obtain 80 mg of a yellow solid. m / z(ES+)(M+H) + = 1147.7.

[0677] Synthesis of compound 99-7 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 mixture was concentrated to obtain 73 mg of crude product as a yellow oily substance. m / z(ES+)(M+H) + = 1047.6.

[0678] Synthesis of LD-99 Compound 99-7 (73 mg, 0.070 mmol, 1.0 eq) and quaternary ammonium sulfonic acid salt B (35 mg, 0.14 mmol, 2.0 eq) were dissolved in DMF (2 mL), and 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. After filtering the reaction mixture, 9.2 mg of a white solid was obtained by preparative purification. LCMS:m / z(ES+)(M+H) + =1284.6, Rt=1.217 min 1 H NMR(400 MHz, DMSO)δ 10.01 (d, J = 38.5 Hz, 1H), 9.24 (s, 1H), 8.26 - 8.16 (m, 1H), 8.13 - 8.01 (m, 1H), 7.80 (t, J = 9.2 Hz, 2H), 7.64 (dd, J = 47.3, 8.1 Hz, 1H), 7.41 - 7.31 (m, 3H), 7.17 (s, 1H), 7.10 (d, J = 4.1 Hz, 2H), 6.54 (s, 1H), 5.44 (d, J = 4.0 Hz, 2H), 5.29 (s, 3H), 5.13 (d, J = 8.2 Hz, 2H), 4.64 (d, J = 27.5 Hz, 2H), 4.36 (d, J = 7.5 Hz, 1H), 4.22 - 4.16 (m, 1H), 3.77 (s, 2H), 3.22 (s, 2H), 3.05 (s, 5H), 2.99 (d, J = 3.8 Hz, 4H), 2.94 (s, 2H), 2.87 (s, 2H), 2.81 (s, 4H), 2.36 (d, J = 18.7 Hz, 5H), 2.19 (dd, J = 13.7, 7.2 Hz, 5H), 2.06 - 1.84 (m, 9H), 1.53 (s, 5H), 1.34 - 1.21 (m, 7H), 0.89 - 0.83 (m, 9H).

[0679] Test Example 18 (Preparation of compound of formula (2)) Binding method 1 (complete reduction, drug-to-antibody ratio 8) The antibody was liquid-exchanged to pH 7.4 phosphate buffer using an ultrafiltration centrifuge tube, and 10 to 15 times the amount of TCEP relative to the molar number of antibodies was added. The mixture was reacted at 37°C for 2 hours to open the disulfide bonds between the antibody chains. After reduction, the antibody was liquid-exchanged to pH 7.4 phosphate buffer using an ultrafiltration centrifuge tube to remove excess TCEP, and 12 to 20 times the amount of Linker-Drug (formula (1) compound, compound MC-GGFG-Exatecan, compound MC-GGFG-Dxd, and compound ELC122 prepared in Examples 1 to 5 and Examples 7 to 17) relative to the molar number of antibodies was added to each, and the mixture was reacted at room temperature for 1 hour. After the reaction was complete, 2 times the amount of cysteine ​​relative to the molar number of Linker-Drugs was added to quench the reaction, and the mixture was liquid-exchanged to pH 5.5 acetate buffer using an ultrafiltration centrifuge tube. The antibody-drug conjugate samples after liquid exchange were filtered through a 0.22 micron sterile filter to prepare for use.

[0680] Binding method 2 (partial reduction, drug-antibody ratio 4) The antibody was liquid-exchanged to pH 7.4 phosphate buffer using an ultrafiltration centrifuge tube, and 2.2 to 2.3 times the amount of TCEP relative to the molar number of antibody molecules was added. The mixture was reacted at 37°C for 2 hours to open the disulfide bonds between the antibody chains. After reduction, 6 to 8 times the amount of Linker-Drug (compound (1) and compound MC-VC-PAB-MMAE prepared in Example 6) relative to the molar number of antibody molecules was added, and the mixture was reacted at room temperature for 1 hour. After the reaction was complete, 2 times the amount of cysteine ​​relative to the molar number of Linker-Drug molecules was added to quench the reaction, and the mixture was liquid-exchanged to pH 5.5 acetate buffer using an ultrafiltration centrifuge tube. The antibody-drug conjugate sample after liquid exchange was filtered through a 0.22 micron sterile filter and prepared for use.

[0681] Binding antibody sequence Anti-Claudin 18.2 humanized monoclonal antibody 9D1 heavy chain amino acid sequence SEQ ID NO:1: QIQLVQSGAEVKKPGASVKISCKASGYSFTDYHMNWVRQAPGKGLEWIGNIDPYYGSPTYNHKFKGRVTLTVDTSTSTAYMELSSLRSEDTAVYYCANYGRGNSFPYWGQG TLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0682] Anti-Claudin 18.2 humanized monoclonal antibody 9D1 light chain amino acid sequence SEQ ID NO:2: DIVMTQSPSSLAVSLGERATINCKSSQSLLNSGNQRNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQNDYSFPFTFGQGTKL EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0683] Farletuzumab, a humanized monoclonal antibody targeting the human folate receptor (FRa). Antibody heavy chain amino acid sequence (SEQ ID NO: 3): EVQLVESGGGVVQPGRSLRLSCSASGFTFSGYGLSWVRQAPGKGLEWVAMISSGGSYTYYADSVKGRFAISRDNAKNTLFLQMDSLRPEDTGVYFCARHGDDPAWFAYWGQG TPVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0684] Farletuzumab, a humanized monoclonal antibody targeting the human folate receptor (FRa). Antibody light chain amino acid sequence (SEQ ID NO: 4): DIQLTQSPSSLSASVGDRVTITCSVSSSISSNNLHWYQQKPGKAPKPWIYGTSNLASGVPSRFSGSGSGTDYTFTISSLQPEDIATYYCQQWSSYPYMYTFGQGTKVE IKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0685] Chromatographic analysis of antibody-drug conjugates For size exclusion chromatography (SEC), the analytical column was Tskgel G3000SWXL (TOSOH, 0008541), and the mobile phase was 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. The detection wavelengths were 280 nm and 366 nm. For hydrophobic interaction chromatography (HIC), the analytical column was Butyl-NPR 4.6 × 100 mm (TOSOH, 0042168), and mobile phase A was 50 mM PB pH 7.0, 1.5 M (NH4)2SO4, while mobile phase B was 50 mM PB pH 7.0, 20% acetonitrile. The separation gradient was from 0% B to 100% B in 2 to 20 minutes, with a column temperature of 30°C and a flow rate of 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 Figures 1A and 1B, Figure 2 includes Figures 2A and 2B, Figure 3 includes Figures 3A, 3B, 3C, 3D, 3E, 3F, 3G, and 3H, Figure 4 includes Figures 4A and 4B, Figure 5 includes Figures 5A and 5B, and Figure 6 includes Figures 6A, 6B, 6C, 6D, 6E, 6F, 6G, and 6H).

[0686] [Table 1-1]

[0687] [Table 1-2]

[0688] Summary: As can be seen from Table 1 and Figures 1-6, the anti-claudin 18.2 humanized monoclonal antibody 9D1 (WO2021160155) was used as a model antibody, and antibody-drug conjugates were prepared by conjugating it with the designed linker drug compound and the control linker drug compound. The linker drug compounds MC-GGFG-Exatecan (ShangHai HaoYuan ChemExpress, catalog number HY-114233), MC-VC-PAB-MMAE (ShangHai HaoYuan ChemExpress, catalog number 646502-53-6), and MC-GGFG-Dxd (Deruxtecan) (ShangHai HaoYuan ChemExpress, catalog number HY-13631E) were purchased. LD-74 was synthesized using the synthesis method of WO2022228493. ELC122 was synthesized using the synthesis method described in WO2021225892.

[0689] Size exclusion chromatography (SEC) analysis of the obtained 9D1 antibody-drug conjugates showed that the SEC purity of the antibody-drug conjugates exceeded 97%, regardless of whether the conjugated drug was Exatecan, SN38, or MMAE. The purity of the antibody-drug conjugate with the control linker LD-27 was only 89.22% (Table 1).

[0690] Hydrophobic interaction chromatography (HIC) analysis of 9D1 antibody-drug conjugates revealed that the HIC retention times of the internal salt linkers designed and invented for the toxins MMAE, SN38, and Exatecan were significantly shorter compared to the 9D1 antibody-drug conjugates of the control linker (excluding LD-74). The retention time of the main peak (drug-antibody ratio 4) of the internal salt linker-MMAE antibody-drug conjugate (here, the antibody-drug conjugate of LD-57) was shortened by approximately 1 minute (16.07 vs 16.99) compared to the control linker-MMAE (MC-VC-PAB-MMAE) antibody-drug conjugate (here, the antibody-drug conjugate of Comparative Example 2), with the retention time being significantly shorter for components with increasing drug-antibody ratios (Table 1, Figure 4). The retention time of the internal salt linker-SN38 antibody-drug conjugate (LD-71 antibody-drug conjugate) was approximately 4 minutes shorter than that of the control linker-SN38 (ELC122, WO2021225892) antibody-drug conjugate (here, the antibody-drug conjugate of Comparative Example 3) (12.93 vs 16.74, Table 1, Figure 5). The HIC retention times of the internal salt linker-Exatecan antibody-drug conjugates of this 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) were 13.12 to 14.17 minutes. This was significantly shorter than the retention times of the control linker-Exatecan (LD-12, LD-27, MC-GGFG-Exatecan) and MC-GGFG-Dxd antibody-drug conjugates without hydrophilic side chains, significantly shorter than the retention time of the LD-73 antibody-drug conjugate with polyethylene glycol (PEG9) hydrophilic side chains (16.00 minutes), and equivalent to the retention time of the LD-74 antibody-drug conjugate with polysarcosine hydrophilic side chains (13.58 minutes) (Table 1, Figure 6). An interesting finding is that, in the case of the internal salt linker-Exatecan, the closer the internal salt with the same structure is to the Exatecan toxin on the linker, the shorter the HIC retention time of the corresponding antibody-drug conjugate.For example, in the case of the sulfonic acid internal salt linker-Exatecan, the HIC retention time of the antibody-drug conjugate corresponding to LD-37 (where the sulfonic acid internal salt is at position Z in formula (1-1), far from Exatecan) was 14.07 minutes, while the retention times of the antibody-drug conjugates corresponding to LD-38 (where the sulfonic acid internal salt is at position Z in formula (1-1-1)), LD-65 (where the sulfonic acid internal salt is at position Z' in the formula), and LD-31(P2) (where the sulfonic acid internal salt is at position Z'' in the formula) were 13.57, 13.46, and 13.40 minutes, respectively, which were significantly shorter than the retention time of 14.07 minutes for the antibody-drug conjugate corresponding to LD-37 (Table 1, Figure 6).

[0691] Hydrophobic interaction chromatography reveals the hydrophobic properties of antibody-drug conjugates, which are primarily due to the hydrophobicity of the bound drug compound. Antibody drugs are highly hydrophobic, resulting in longer retention times in hydrophobic interaction chromatography, while antibody-drug conjugates are easily excreted (cleared) from the body due to their low water solubility and stability. The extremely strong water solubility of the internal salt linker significantly masks the hydrophobic properties of the drug compound, improving the water solubility and stability of the antibody-drug conjugate and shortening its retention time in hydrophobic interaction chromatography.

[0692] Furthermore, conjugates were prepared of the internal salt linker-Exatecan (LD-38) with the anti-human folate receptor (FRa) humanized monoclonal antibody farletuzumab and an IgG1 isotype control (Biointron, catalog no. B117901) monoclonal antibody. Their SEC purity was >96%, and their HIC retention time was approximately 13 minutes (Table 1). This is comparable to the retention time of the corresponding 9D1 antibody-drug conjugate.

[0693] The 9D1 antibody-drug conjugates (i.e., the compounds shown in formula (2)) in the following test examples were all prepared in Test Example 18.

[0694] Test Example 19 (Drug-Antibody Ratio (DAR)) Sample processing: The 9D1-LD-38 prepared in Test Example 5a was diluted to 1 mg / mL with 0.1 M Tris-HCl, and 5 μL of 0.5 M DTT was added per 100 μg of 9D1-LD-38. The mixture was treated in a 37°C water bath for 30 minutes, and then 1% (v / v) 10% TFA was added to stop the reaction.

[0695] LC-MS parameters: Mobile phase A was an aqueous solution of 0.10% TFA and 0.02% FA, and mobile phase B was an acetonitrile solution of 0.08% FA and 0.02% TFA. The separation gradient was adjusted by changing the concentration of mobile phase B from 20% to 32% over 2-4 minutes, and from 32% to 45% over 4-12 minutes. The flow rate was 0.4 mL / min. A BioResolve™ RP mAb Polyphenyl Column (2.7 μm, 2.1 × 150 mm) was used as the chromatography column, and the column temperature was 70°C.

[0696] The mass spectrometer was a Themo Q Exactive Plus with a spray voltage of 3.8kV, a scan range of 500-3500m / z, and full scan in positive ion mode.

[0697] RP-HPLC parameters: Mobile phase A was a 0.1% TFA aqueous solution, and mobile phase B was a 0.1% TFA acetonitrile solution. The separation gradient was adjusted by changing the concentration of mobile phase B from 30% to 50% over 5 to 25 minutes. The flow rate was 0.6 mL / min, the chromatography column was a Proteomix RP-1000 (5 μm, 4.6 × 100 mm), and the column temperature was 60°C. The results are shown in Table 2 and Figures 7-9.

[0698] [Table 2]

[0699] Summary: As can be seen from Table 2 and Figures 7-9, 9D1-LD-38 is pretreated to maintain glycosylation in the reduced state because it is bound to cysteine. The results showed that the light chain and heavy chain each have one main component. According to molecular weight identification by mass spectrometry, the main binding form of the light chain was one LD-38 molecule, and the main binding form of the heavy chain was three LD-38 molecules. The results of molecular weight deconvolution are shown in Figures 7 and 8, and the results are consistent with the theoretical molecular weight. Furthermore, partial ring opening of the LD-38 molecule and the existence of heteromorphs with different numbers of bonds in the heavy chain were revealed. The DAR value of 9D1-LD-38 was calculated to be 7.8 by area normalization of the liquid phase spectrum (Figure 9), which corresponds to a theoretical binding drug-antibody ratio of 8.

[0700] Test Example 20 (In vitro tumor cell killing activity) 1. Human claudin 18.2-positive MC38-hClaudin18.2 cell line In vitro sterilization was performed on MC38 cell lines stably expressing human claudin 18.2 using the 9D1 antibody-drug conjugates prepared in the aforementioned test examples and comparative examples. MC38-hClaudin18.2 cells were inoculated at a rate of 3000 cells / well into 96-well cell culture plates and cultured overnight. The following day, serially diluted 9D1 ADCs were added, and the cells were cultured for 48-72 hours. Cell viability was calculated by detecting the fluorescence value of each experimental well using Alamar Blue reagent (ThermoFisher, catalog number DAL1100).

[0701] MC38-hClaudin18.2 cells were cultured in adherent culture in a 37°C, 5% CO2 incubator using RPMI 1640 medium containing 10% FBS and 1 μg / mL puromycin. Once cell confluence exceeded 80%, the cells were subcultured every 3 days at a ratio of 1:10 to 1:20.

[0702] MC38-hClaudin18.2 cells were digested with recombinase digestate (TrypLE™ Express Enzyme). After counting the number of cells, an appropriate amount was collected and resuspended in RPMI 1640 + 10% FBS, and the cell density was set to 3 × 10⁶. 4 The reagent was adjusted to cells / mL and inoculated at 100 μL / well into a 96-well cell culture plate, where it was incubated overnight at 37°C in a 5% CO2 incubator. The following day, the reagent was serially diluted 10-fold to a total of 7-8 concentrations. A blank control with a sample concentration of 0 was also prepared. The diluted samples were added at 100 μL / well to a 96-well cell culture plate containing cells 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. Fluorescence signal intensity was detected using a multi-functional microplate reader at an excitation wavelength of 560 nm and an fluorescence wavelength of 590 nm.

[0703] The cell viability of the blank control was set to 100%, and the cell viability (%) of the experimental wells was calculated according to the following formula. Cell viability (%) = (Fluorescence value of experimental well / Fluorescence value of blank control well) × 100%

[0704] The mean of the measured cell viability was used as the vertical axis, and cell proliferation inhibition curves were obtained using the sigmoidal dose-response (Variable Slope) method (GraphPad Prism software, GraphPad Software, San Diego, California) for nonlinear regression. The corresponding half-maximal inhibitory concentration (IC50) values ​​were calculated. The results are shown in Table 3 and Figure 10 (Figures 10A, 10B, 10C, 10D, and 10E).

[0705] [Table 3]

[0706] Summary: As can be seen from Table 3 and Figure 10, all 9D1 antibody-drug conjugates specifically killed MC38-hClaudin18.2 with Exatecan toxin, compared to drug conjugates of IgG1 isotype control antibodies (Isotype IgG1-LD-12, Isotype IgG1-LD-38), with IC50 values ​​ranging from 0.7 to 11.03 nM. Among these, 9D1-GGFG-Exatecan had the worst pharmacodynamic activity, with an IC50 of 11.03 nM. The IC50 values ​​of 9D1 antibody-drug conjugates with internal hydrophilic side chains ranged from 0.8 to 4.4 nM. With the exception of 9D1-LD-31-P1, their activity was significantly stronger than that of Exatecan small molecule toxin (IC50 value 4.10 nM).

[0707] Regarding MMAE toxins, the IC50 value of the drug conjugate of the IgG1 isotype control antibody (Isotype IgG1-VC-PAB-MMAE) was 41.3 nM for MC38-hClaudin 18.2, while 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.

[0708] 2. Human claudin 18.2-positive NUGC4-hClaudin18.2 cell line Human gastric cancer cell line NUGC4 (NUGC4-hClaudin18.2), which stably expresses human claudin 18.2, was cultured in adherent culture in a 37°C, 5% CO2 incubator using RPMI 1640 medium containing 10% FBS and 1 μg / mL puromycin. When cell confluence exceeded 80%, the cells were subcultured every 3-4 days at a ratio of 1:3 to 1:10.

[0709] NUGC4-hClaudin18.2 cells were digested with recombinase digestate (TrypLE™ Express Enzyme). After counting the number of cells, an appropriate amount was collected and resuspended in RPMI 1640 + 10% FBS, and the cell density was set to 3 × 10⁶. 4The sample was adjusted to cells / mL and inoculated at 100 μL / well into a 96-well cell culture plate. The cells were incubated overnight at 37°C in a 5% CO2 incubator. The following day, 9D1 ADC and small molecule toxins were serially diluted five-fold from 1000 nM (final concentration 500 nM) to a total of 7-8 concentrations. A negative control with a sample concentration of 0 and a blank control with only culture medium were also prepared. The diluted samples were added to 96-well cell culture plates containing cells at 100 μL / well (overlapping wells) and incubated at 37°C in a 5% CO2 incubator for 120 hours. 20 μL of CCK-8 detection reagent (Tojin Chemical Co., Ltd., catalog number CK04) was added to each well and incubated at 37°C for 3-4 hours. The OD values ​​of each well in the ELISA plate were measured using a multifunctional microplate reader at a measurement wavelength of 450 nm and a reference wavelength of 630 nm. The OD values ​​for each well ranged from OD450 nm to OD630 nm. The cell viability of the negative control was set to 100%, and the cell viability (%) of the experimental wells was calculated according to the following formula. 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%

[0710] The sigmoidal dose-response (Variable Slope) method (GraphPad Prism software, GraphPad Software, San Diego, California) was used for nonlinear regression to calculate the corresponding half-maximal inhibitory concentration (IC50). The results are shown in Table 4.

[0711] [Table 4]

[0712] Summary: As can be seen from Table 4, for Exatecan toxin, all 9D1 antibody-drug conjugates specifically killed NUGC4-hClaudin18.2 with IC50 values ​​of 0.18–1.27 nM, compared to drug conjugates of IgG1 isotype control antibody (Isotype IgG1-LD-38, IC50 141.3 nM). The IC50 values ​​of 9D1 antibody-drug conjugates with internal salt hydrophilic side chains (0.55–1.00 nM) were all superior to those of antibody-drug conjugates with polyethylene glycol (PEG, 9D1-LD-73, IC50 1.27 nM) and polysarcosine (PSAR, 9D1-LD-74, IC50 1.16 nM) hydrophilic side chains.

[0713] Regarding MMAE toxins, the IC50 values ​​of the 9D1-LD-57 antibody-drug conjugate with an internally salty hydrophilic side chain and the control antibody-drug conjugate 9D1-vc-PAB-MMAE were equivalent, at 0.05 nM and 0.06 nM, respectively.

[0714] 3. Human claudin 18.2-positive HEK293-hClaudin18.2 cell line HEK293-hClaudin18.2 cells were cultured in adherent culture in a 37°C, 5% CO2 incubator using DMEM medium containing 10% FBS and 250 μg / mL of G418. Once cell confluence exceeded 80%, the cells were subcultured every 3 days at a ratio of 1:20 to 1:40.

[0715] HEK293-hClaudin18.2 cells were digested with recombinase digestate (TrypLE™ Express Enzyme), the cell count was taken, and an appropriate amount of cells were collected and resuspended in DMEM + 10% FBS. The cell density was set to 2 × 10⁻⁶. 4The sample was adjusted to cells / mL and inoculated at 100 μL / well into a 96-well cell culture plate. The cells were incubated overnight at 37°C in a 5% CO2 incubator. The following day, 9D1 ADC and small molecule toxins were serially diluted 10-fold from 1000 nM (final concentration 500 nM) to a total of 7-8 concentrations. A negative control with a sample concentration of 0 and a blank control with no cells and only the same volume (200 μL / well) of culture medium were also prepared. The diluted samples were added at 100 μL / well (overlapping wells) to a 96-well cell culture plate containing cells and incubated for 72 hours at 37°C in a 5% CO2 incubator. Cell activity was detected and data processed using the AlamarBlue detection method described above, and the IC50 value was calculated. The results are shown in Table 5.

[0716] [Table 5]

[0717] Summary: As can be seen from Table 5, regarding Exatecan toxin, compared to the drug conjugate of the IgG1 isotype control antibody (Isotype IgG1-LD-38, IC50 688.2 nM), all 9D1 antibody-drug conjugates with an internal salt hydrophilic side chain specifically killed HEK293-hClaudin18.2, with IC50 values ​​ranging from 0.09 to 1.00 nM, while the IC50 of small molecule Exatecan was 0.51 nM.

[0718] For the SN38 toxin, the 9D1 antibody-drug conjugate was shown to specifically kill HEK293-hClaudin18.2 compared to the drug conjugate of the IgG1 isotype control antibody (Isotype IgG1-ELC122, no killing). Killing by the 9D1 antibody-drug conjugate with an internal salt hydrophilic side chain (9D1-LD-57) was slightly stronger than that of the control (9D1-ELC122), with IC50 values ​​of 0.40 nM and 0.63 nM, respectively.

[0719] Test Example 21 (In vivo tumor treatment effect) 1. Human claudin 18.2-positive MC38-hClaudin18.2 cell line mouse transplant tumor model C57BL / 6 mice (female, 5-6 weeks old) were purchased from Shanghai Lingchang Co., Ltd. The animals were housed in laminar flow racks in the SPF animal room, with 5 animals per cage, under 12-hour alternating lighting. The animals had free access to water and food. Mouse colorectal cancer cell line MC38-Claudin18.2, stably expressing human claudin 18.2, was cultured in T75 square flasks in RPMI 1640 medium containing 10% FBS (Fetal Bovine Serum) and incubated at 37°C in a 5% CO2 incubator. Cells were passaged approximately every 3 days. MC38h Claudin18.2 cells in the logarithmic growth phase were harvested, centrifuged at 1000 rpm for 5 minutes, and resuspended in PBS. 2 × 10 6 The cells were uniformly mixed with a matrix gel in a 1:1 volume ratio and inoculated subcutaneously into C57BL / 6 mice. The tumors were 100-150 mm in size. 3 Once the tumors reached a certain volume, the mice were divided into groups and administered drugs (5-6 mice / group). The size of the tumors was measured twice a week, and the tumor volume (TV, mm) was measured. 3 The coefficients were calculated and the tumor growth curve was plotted. TV (mm 3 )=(L×W 2 ) / 2, where L represents the length of the tumor and W represents the width of the tumor. Tumor inhibition rate (%) = (TV control group - TV treatment group) × 100 / TV control group

[0720] Mouse tumor curves were plotted in XY mode using GraphPad Prism software (GraphPad Software, San Diego, California), with the number of days after the first dose on the x-axis and tumor volume on the y-axis. Two-way ANOVA was used to analyze whether there were statistically significant differences between each group and the Hibicul-controlled PBS group (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001).

[0721] Table 6 shows the treatment regimens of the 9D1 antibody-drug conjugate conjugate conjugate conjugate conjugate conjugate in the MC38-hClaudin18.2 mouse subcutaneous tumor model, and the tumor therapeutic effects and corresponding tumor growth curves are shown in Table 6 and Figure 11 (Figure 11 includes Figures 11A, 11B, and 11C).

[0722] [Table 6]

[0723] Summary: As can be seen from Table 6 and Figure 11, under a single dose of 10 mg / kg, tumor growth was slowed with control antibody-drug conjugates (9D1-ELC122, 9D1-GGFG-Exatecan, 9D1-LD-12) (Figure 11A), and the tumor inhibition rates (TGI) at the end of the experiment were 39.82%, 48.34%, and 48.63%, respectively. Antibody-drug conjugates with internal bases in the linker (9D1-LD-31-P2 and 9D1-LD-38) showed superior tumor therapeutic activity, with TGIs of 80.0% and 80.6%, respectively (Figure 11B).

[0724] When 10 mg / kg was administered once a week for a total of two doses, the tumor inhibition rate for the control antibody-drug conjugate 9D1-GGFG-Dxd was 50.0% at the end of the experiment, while tumors treated with 9D1-LD-38 almost completely disappeared (tumor inhibition rate 98.0%). The isotype control antibody-drug conjugate (IgG1 Isotype-LD-38) showed a slight tumor inhibitory effect (Table 6).

[0725] To further compare and differentiate the tumor therapeutic effects of other 9D1 antibody-drug conjugates containing internal bases, low doses (5 mg / kg) were selected for the efficacy experiments, and 9D1-LD-38 was set as an internal reference. Under low-dose (5 mg / kg) conditions, the isotype control antibody-drug conjugate (IgG1 Isotype-LD-38) did not show any significant tumor therapeutic effect. The tumor therapeutic effect of 9D1-LD-38 was remarkable, with a TGI of 39.2%. Other 9D1 antibody-drug conjugates containing internal bases in the linker (9D1-LD-65, 9D1-LD-66, and 9D1-LD-72) showed superior tumor therapeutic effects (Figure 11C), with TGIs reaching 71.7%, 48.0%, and 42.3%, respectively. In particular, 9D1-LD-65 exhibited the most superior tumor inhibitory effect, with significantly better tumor growth inhibition curves (Figure 11C) and TGI than other reported 9D1 antibody-drug conjugates containing other water-soluble groups in the linker, such as 9D1-LD-73 (TGI 47.0%) control containing polyethylene glycol (PEG) water-soluble group and 9D1-LD-74 (TGI 56.5%) control containing polysarcosine (PSAR) water-soluble group.

[0726] As described above, 9D1 antibody-drug conjugates containing an internal salt water-soluble group in the linker (9D1-LD-31-P2, 9D1-LD-38, 9D1-LD-65, 9D1-LD-66, and 9D1-LD-72) exhibited significantly superior efficacy compared to conventional linker-antibody-drug conjugates without water-soluble groups, such as 9D1-ELC122, 9D1-GGFG-Exatecan, and 9D1-LD12. In particular, 9D1-LD-65, 9D1-LD-66, and 9D1-LD-72 showed tumor therapeutic efficacy equivalent to or better than 9D1 antibody-drug conjugates containing other types of water-soluble groups in the linker (such as 9D1-LD-73 containing PEG and 9D1-LD-74 containing PSAR).

[0727] 2. Human claudin 18.2-positive NUGC4-hClaudin18.2 cell line mouse transplant tumor model NUGC4-hClaudin18.2, a positive gastric cancer cell line 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 in a 37°C, 5% CO2 incubator. Every 4-5 days, after the cells had grown sufficiently (reached 100% confluence), they were subcultured in a separate flask, and tumor cells in the logarithmic growth phase were used for in vivo tumor inoculation.

[0728] NUGC4-hClaudin18.2 tumor cells were sampled in PBS:Matrigel (mixture in a 1:1 volume ratio) at a rate of 5 × 10⁻¹⁴. 7 The concentration was adjusted to 100 μL / mL and subcutaneously inoculated into the right flank of BALB / c nu / nu mice (female, 6-7 weeks old) at a dose of 100 μL / mice. The tumor cell inoculation amount was 5 × 10⁻⁶. 6 It was one animal. The tumor was approximately 150 mm. 3 Once the tumors had grown to a certain volume, the mice were divided into groups and administered the treatment. The group divisions and administration regimens are shown in Table 7. Tumor size was measured twice a week, and tumor volume (TV, mm) was measured. 3 The coefficients were calculated and the tumor growth curve was plotted. TV (mm 3 )=(L×W 2 ) / 2, where L represents the length of the tumor and W represents the width of the tumor.

[0729] Using GraphPad Prism software (GraphPad Software, San Diego, California), mouse tumor curves were plotted in XY mode with the number of days after the first dose on the x-axis and tumor volume on the y-axis. The results are shown in Figure 12.

[0730] [Table 7]

[0731] Summary: As can be seen from Table 7 and Figure 12, under the condition of administration of 5 mg / kg once a week for three times, 9D1-LD-38, which contains an internal salt water solubility promoting group in its linker, showed the best therapeutic effect and was significantly superior to 9D1-ECL122 and 9D1-GGFG-Exatecan. Tumors treated with 9D1-LD-38 almost disappeared 24 days after treatment, whereas in the 9D1-GGFG-Exatecan treatment group, tumor growth was inhibited but no tumor reduction was observed. 9D1-ECL122 showed only slight tumor inhibitory effect, which is thought to be because the toxin of 9D1-ECL122 is the less toxic SN38.

[0732] 3. Folate receptor-positive OVCAR-3 cell transplantation tumor model in mice Human ovarian cancer tumor cells expressing folate receptor (FRa) (OVCAR-3) were cultured in RPMI-1640 medium containing inactivated 20% fetal bovine serum, 100 U / ml penicillin, and 100 μg / ml streptomycin in a 37°C, 5% CO2 incubator. Every 4-5 days, after the cells had grown sufficiently, they were subcultured in a separate flask, and tumor cells in the logarithmic growth phase were used for in vivo tumor inoculation.

[0733] OVCAR3 tumor cells were mixed in PBS:Matrigel (1:1 volume ratio) at a rate of 1 × 10⁶ 8 The concentration was adjusted to 1 / mL, and 100uL / mice were subcutaneously inoculated into the right flank of M-NSG experimental mice, with a tumor cell inoculation amount of 1 × 10⁶. 7 There were / animals.

[0734] The tumor was approximately 150 mm in size. 3 Once the tumors had grown to a certain volume, the mice were divided into groups (5 mice per group) and administered the treatment. The control group received PBS, while the treatment group received Farletuzumab-LD-38 antibody-drug conjugate, a humanized monoclonal antibody against the human folate receptor (FRa). The dose was 5 mg / kg, administered as a single dose. Tumor size was measured twice a week, and tumor volume (TV, mm) was monitored. 3 The coefficients were calculated and the tumor growth curve was plotted. TV (mm3 )=(L×W 2 ) / 2, where L represents the length of the tumor and W represents the width of the tumor. Tumor inhibition rate (%) = (TV control group - TV treatment group) × 100 / TV control group

[0735] Mouse tumor curves were plotted in XY mode using GraphPad Prism software (GraphPad Software, San Diego, California), with the number of days after the first dose on the x-axis and tumor volume on the y-axis. Two-way ANOVA was used to analyze whether there were statistically significant differences between each group and the Hibicul-controlled PBS group (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001). The results are shown in Figure 13.

[0736] Summary: As shown in Figure 13, the Farletuzumab-LD-38 antibody-drug conjugate showed tumor growth inhibition one week after a single 5 mg / kg dose compared to the control group, and subsequent tumor reduction was observed. By day 24, the tumor growth inhibitory rate (TGI) reached 93.5%, showing a significant difference compared to the control group.

[0737] 4. Folate receptor-positive IGROV1 cell transplantation tumor model in mice 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 in a 37°C, 5% CO2 incubator. Every 3-4 days, after the cells had grown sufficiently, they were subcultured in a separate flask, and tumor cells in the logarithmic growth phase were used for in vivo tumor inoculation.

[0738] IGROV1 tumor cells were incubated in PBS at a rate of 1 × 10⁶ 8 The concentration was adjusted to 1 / ml, and 100 μL / mice was subcutaneously inoculated into the right flank of BALB / c nude experimental mice, with a tumor cell inoculation amount of 1 × 10⁶. 7 There were / animals.

[0739] The average tumor volume is approximately 150 mm². 3At the point when the target was reached, the animals were divided into groups and administered the treatment (the day of group division was recorded as D0). Each model was divided into two groups, with five animals in each group. The control group was administered PBS, and the treatment group was administered Farletuzumab-LD-38 antibody-drug conjugate, a humanized monoclonal antibody against the human folate receptor (FRa). The dose was 5 mg / kg, administered twice a week. Tumor size was measured twice a week, and tumor volume (TV, mm) was measured. 3 The coefficients were calculated and the tumor growth curve was plotted. TV (mm 3 )=(L×W 2 ) / 2, where L represents the length of the tumor and W represents the width of the tumor. Tumor inhibition rate (%) = (TV control group - TV treatment group) × 100 / TV control group

[0740] Mouse tumor curves were plotted in XY mode using GraphPad Prism software (GraphPad Software, San Diego, California), with the number of days after the first dose on the x-axis and tumor volume on the y-axis. Two-way ANOVA was used to analyze whether there were statistically significant differences between each group and the Hibicul-controlled PBS group (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001).

[0741] The results are shown in Figure 14. Farletuzumab-LD-38 antibody-drug conjugate, administered at a dose of 5 mg / kg once weekly for two doses, showed rapid tumor reduction compared to the control group, with a tumor inhibition rate (TGI) reaching 100% on day 30, and its efficacy was maintained until day 50 after administration.

[0742] Study Example 22: In vivo pharmacokinetics in rats For the study, three male SD rats were selected and randomly divided into three groups. Each rat received a single intravenous injection of either 9D1-LD-38 (prepared in the test example) or 9D1-GGFG-Exatecan (prepared in the comparative example) at a dose of 10 mg / kg. Whole blood samples were collected from each 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. Serum and plasma were separated. The concentrations of Total Ab (total antibody) and ADC (bound antibody) of each test in the serum samples were analyzed by ELISA. Pharmacokinetic parameters for each animal were calculated using the PK Solver non-compartment model (NCA).

[0743] Measurement method for total antibody and conjugated antibody: Blood concentrations of total antibody and conjugated antibody were measured using the ELISA method. The specific method is as follows.

[0744] a) Anti-human IgG(Fab)2 was diluted to 1 μg / mL with PBS, coated at 100 μL / well in a 96-well ELISA plate, and left overnight at 4°C.

[0745] b) The plates were washed three times and blocked with 3% BSA-PBST at 300 μL / well for 2 hours at room temperature.

[0746] c) The plate was washed three times, and 9D1-LD-38 / 9D1-GGFG-Exatecan was diluted 2-fold with rat blank serum to a total of 7 concentrations, with a starting concentration of 1 μg / mL. The "0" point was set as rat blank serum. Additionally, samples were diluted appropriately using rat blank serum at different time points after drug administration. All of the above samples were diluted 50-fold (9D1-GGFG-Dxd) and 100-fold (9D1-LD-38) with 1% BSA-PBST, added to an ELISA plate at 100 μL / well, and incubated at room temperature for 2 hours.

[0747] d) The plate was washed six times. Detection of total antibodies: HRP-goat anti-human IgGFC was diluted 1:20000 with 1% BSA-PBST, added to an ELISA plate at a rate of 100 μL / well, and incubated at room temperature for 1 hour.

[0748] ADC measurement: Anti-Exatecan antibody was diluted to 0.5 μg / mL using 1% BSA-PBST, added to an ELISA plate at 100 μL / well, and incubated at room temperature for 1 hour. The plate was washed 6 times, and HRP goat anti-mouse IgG (H+L) was diluted 1:20000 using 1% BSA-PBST, added to an ELISA plate at 100 μL / well, and incubated at room temperature for 1 hour.

[0749] The plate was washed six times, and 100 μL / well of TMB was added to the ELISA plate. The mixture was allowed to develop color in the dark at room temperature for approximately 10 minutes. A stop solution was added at a rate of 50 μL / well. The results were detected using a microplate reader (wavelength 450 / 630 nm) and are shown in Table 8 and Figure 15.

[0750] [Table 8]

[0751] Summary: As can be seen from Table 8 and Figure 15, a single intravenous injection of 10 mg / kg into rats resulted in a rapid decrease in serum drug concentrations of 9D1-GGFG-Exatecan total antibody and conjugated antibody (ADC) (Figure 15), with short half-lives of 65.1 hours and 57.8 hours, respectively. The ratio of the AUC of the conjugated antibody to the AUC of the total antibody was 0.67.

[0752] The serum drug concentrations of 9D1-LD-38 total antibody and conjugated antibody decreased relatively slowly (Figure 15), with half-lives of 145.4 hours and 119.8 hours, respectively, and the ratio of the AUC of the conjugated antibody to the AUC of the total antibody was 0.78.

[0753] The half-life of the 9D1-LD-38 conjugated antibody is twice that of 9D1-GGFG-Exatecan, and the AUC of the 9D1-LD-38 conjugated antibody is 1.3 times that of 9D1-GGFG-Exatecan. The ratio of the AUC of the 9D1-LD-38 conjugated antibody to the AUC of the total antibody is higher than the ratio of the AUC of 9D1-GGFG-Exatecan (0.78 vs 0.67), indicating that 9D1-LD-38 is more stable and that small molecules do not easily detach.

[0754] As described above, 9D1-LD-38, which contains an internal salt water solubility-enhancing group in its linker, exhibited significantly weaker drug clearance in rats compared to 9D1-GGFG-Exatecan, which does not contain a water solubility-enhancing group in its linker. Furthermore, 9D1-LD-38 had a significantly longer half-life, resulting in a substantial increase in drug exposure. Its in vivo stability was also superior, as evidenced by the higher ratio of AUC of bound antibody to AUC of total antibody.

[0755] Test Example 23 (In vitro cell-killing effects of 9D1-LD-38, 9D1-LD-82, 9D1-LD-88, and 9D1-LD-89) MC38-hClaudin18.2 cells were cultured in adherent culture in a 37°C, 5% CO2 incubator using RPMI 1640 (Thermo, catalog number: 11875-085) medium containing 10% FBS (Thermo, catalog number: 10099-141C) supplemented with 1 μg / mL puromycin (Thermo, catalog number: A11138-03). Once cell confluence exceeded 80%, the cells were subcultured every 3 days at a ratio of 1:10 to 1:20. MC38-hClaudin18.2 cells were digested with recombinase digestate (TrypLE™ Express Enzyme (Thermo, catalog number: 12605028)). After counting the number of cells, an appropriate amount of cells was collected, resuspended in RPMI 1640 + 10% FBS, and the cell density was increased to 1.5 × 10⁶. 4The samples were adjusted to cells / mL and inoculated at 100 μL / well into 96-well cell culture plates. They were incubated overnight at 37°C in a 5% CO2 incubator. The following day, 9D1-LD-38, 9D1-LD-82, 9D1-LD-88, and 9D1-LD-89 were serially diluted 10-fold from a 500 nM molar concentration to a total of eight concentrations. A negative control with a sample concentration of 0 and a blank control with no cells and only the same volume (200 μL / well) of culture medium were also prepared. The diluted samples were added at 100 μL / well (overlapping wells) to 96-well cell culture plates containing cells and incubated at 37°C in a 5% CO2 incubator for 72 hours. 20 μL of AlamarBlue (Thermo, catalog number: DAL1100) detection reagent was added to each well and incubated at 37°C for 6 hours. Fluorescence signal intensity was measured using a multifunctional microplate reader (Molecular Devices, M5) at an excitation wavelength of 560 nm and an fluorescence wavelength of 590 nm. The cell viability of the negative control was set to 100%, and the cell viability (%) of the experimental wells was calculated according to the following formula. 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%

[0756] Using the logarithm of ADC concentration on the x-axis and cell viability on the y-axis, a cell proliferation inhibition curve was obtained using the sigmoidal dose-response (Variable Slope) method (GraphPad Prism software, GraphPad Software, San Diego, California) for nonlinear regression, and is shown in Figure 16.

[0757] Summary: As can be seen in Figure 16, 9D1-LD-38, 9D1-LD-82, 9D1-LD-88, and 9D1-LD-89 all dose-dependently inhibited the proliferation of MC38-hClaudin18.2 cells. The inhibitory effects of 9D1-LD-38, 9D1-LD-88, and 9D1-LD-89 were equivalent (IC50 values ​​of 0.632, 1.458, and 0.757 nM, respectively), while 9D1-LD-82 was slightly weaker (IC50 value of 8.013 nM).

[0758] Test Example 24 (Induction of immunogenic cell death in B16F10-hClaudin18.2 cells) B16F10-hClaudin18.2 cells were revived and cultured in DMEM (Thermo, catalog number: 10569044) + 10% FBS (Thermo, catalog number: 10099-141C) + 0.5 μg / mL puromycin medium in a 37°C, 5% CO2 incubator. When cell confluence exceeded 80%, the cells were subcultured at a ratio of 1:10 to 1:20 every 3-4 days.

[0759] Once the cells have grown to approximately 80% confluence, they are digested with recombinase digestate (TrypLE™ Express Enzyme, Thermo, catalog number: 12605028), resuspended in DMEM + 10% FBS, an appropriate amount of cells is harvested, the cells are counted, and the cell density is set to 1 × 10⁻⁶. 4The ADC drug was adjusted to cells / mL and inoculated at 100 μL / well into a 96-well cell plate. The cells were incubated overnight at 37°C in a 5% CO2 incubator. The following day, the ADC drug was diluted to a final concentration of 750 nM in DMEM + 10% FBS medium, 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 prepared. The diluted samples were added at 100 μL / well to a 96-well cell culture plate containing cells and incubated for 48 hours at 37°C in a 5% CO2 incubator. After removing the 96-well cell culture plate from the incubator and digesting it with Accutase (Thermo, catalog number: 00-4555-56), the plates were washed once with 1% FBS PBS. 100 μl / well of anti-calreticulin-AlexaFluor647 antibody (Abcam, catalog number: AB196159, diluted 1:100 in 1% FBS PBS) was added, and the plates were incubated in the dark at room temperature for 30 minutes. After washing twice with 1% FBS PBS, 2 μl of PI dye (Thermo, catalog number: 00-4555-56) was added to each sample, and the plates were incubated in the dark at room temperature for 10 minutes before being placed in a machine for detection by FACS (Bechman, CytoFLEX). Figure 17 was plotted based on the ratio of calreticulin (+) / PI (-) cells.

[0760] Summary: As can be seen in Figure 17, when the ADC concentration was 750 nM, the calreticulin (+) / PI (-) cell ratio in the 9D1-LD-38 / 9D1-GGFG-Dxd treatment group was significantly higher than that of the control ADC, with the 9D1-LD-38 group having a higher ratio than the 9D1-GGFG-Dxd group. When Dxd was added to the small molecule drug at a concentration of 10 nM, the calreticulin (+) / PI (-) cell ratio increased significantly, with Exatecan (MCE, catalog number: HY-13631A) > Dxd (MCE, catalog number: HY-13631D). The results indicate that 9D1-LD-38 can induce more potent immunogenic cell death compared to 9D1-GGFG-Dxd, providing a rationale for its use in combination with immune checkpoint inhibitors.

[0761] Test Example 25 (Bystander-killing effect on HEK293-hClaudin18.2 / HEK293 cells) AlarmaBlue detection method: hClaudin18.2-positive HEK293-hClaudin18.2 cells and hClaudin18.2-negative HEK293 cells were resuscitated and cultured in DMEM+10%FBS+250μg / mL G418 and DMEM+10%FBS medium, respectively, and adherently cultured in a 37°C, 5% CO2 incubator. When cell confluence exceeded 80%, the cells were subcultured at a ratio of 1:10 to 1:20 every 3-4 days.

[0762] Once the cells have grown to approximately 80% confluence, negative and positive cells are digested with recombinase digestate (TrypLE™ Express Enzyme), the cells are counted, an appropriate amount of cell suspension is centrifuged, and the cells are resuspended in DMEM + 10% FBS to a cell density of 5 × 10⁶. 3 The concentration was adjusted to cells / mL. Both positive and negative cell populations were inoculated into 96-well cell culture plates at 100 μL / well. A mixture of positive and negative cells was uniformly mixed in a 1:1 ratio and inoculated into 96-well cell culture plates at 200 μL / well, and incubated overnight in a 37°C, 5% CO2 incubator. The following day, the ADC drug was diluted to a final concentration of 0.5 nM in DMEM + 10% FBS medium, and a blank control with a sample concentration of 0 was also prepared. 100 μL of medium was discarded from each well of the mixed positive and negative cell population, and the diluted ADC drug was added to the 96-well cell culture plate containing the cells at 100 μL / well, and incubated for 120 hours in a 37°C, 5% CO2 incubator. A 96-well cell culture plate was removed from the incubator, and 20 μL of AlamarBlue (Thermo, catalog number: DAL1100) detection reagent was added to each well. The plate was incubated at 37°C for 4 hours. Fluorescence signal intensity was detected using a multi-functional microplate reader at an excitation wavelength of 560 nm and an fluorescence wavelength of 590 nm.

[0763] The cell viability of the blank control was set to 100%, and the cell viability (%) of the experimental wells was calculated according to the following formula. Cell viability (%) = (Fluorescence value of experimental well / Fluorescence value of blank control well) × 100%

[0764] FACS detection method: HEK293 cell labeling: After counting HEK293 cells in the logarithmic growth phase, an appropriate amount of cell suspension is centrifuged, washed twice with PBS, and the cell density is reduced to 5 × 10⁻⁶. 6 The cells / mL were adjusted, and an appropriate amount of CFSE (eBioscience, 65-0850-84) was added to a final concentration of 0.5 nM. The mixture was immediately homogenized and incubated at room temperature in the dark for 10 minutes. Then, 4-5 times the volume of pre-cooled DMEM+10%FBS was added, and the reaction was stopped by incubation on ice for 5 minutes. After washing twice with pre-cooled DMEM+10%FBS, the cell density was increased to 2 × 10⁶ in DMEM+10%FBS medium. 4 The concentration was adjusted to 1 / mL.

[0765] Co-incubation and detection: HEK293-hClaudin18.2-positive cells were digested with recombinase digestate (TrypLE™ Express Enzyme). After counting the cells, an appropriate amount of cell suspension was centrifuged and resuspended in DMEM + 10% FBS to a cell density of 2 × 10⁶. 4The concentration was adjusted to cells / mL. CFSE-labeled HEK293 cells and HEK293-hClaudin18.2 cells were homogeneously mixed in equal volumes and inoculated into 96-well cell culture plates at 100 μL / well. The mixture was 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 medium, and a blank control with a sample concentration of 0 was prepared. 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, the cells were digested with recombinase digestant, and washed once with 1% FBS PBS. Each sample was resuspended in 250 μL of 1% FBS PBS, and the total number of cells in each sample was calculated by counting the cells. 100 μL was collected from each sample, and the ratio of hClaudin18.2-positive to hClaudin18.2-negative cells was measured by FACS detection. The number of hClaudin18.2-positive and hClaudin18.2-negative cells in each sample was calculated based on the total number of cells.

[0766] Summary: ADC drugs were co-cultured for 5 days with HEK293, HEK293-hClaudin18.2, and HEK293+HEK293-hClaudin18.2 mixed culture cells, and cell viability was measured by the AlarmaBlue method. The results are shown in Figure 18. Figure 18 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, all four ADC drugs did not have a killing effect on negative HEK293 cells, but 9D1-LD-38 and 9D1-GGFG-Dxd all had killing effects on positive HEK293-hClaudin18.2 cells compared to the control (Isotype IgG1) ADC, and the killing effects of these two were equivalent. Compared to the control (Isotype IgG1) ADC, 9D1-LD-38 and 9D1-GGFG-Dxd exhibited killing activity against both positive and negative mixed cells, with 9D1-LD38 being more potent than 9D1-GGFG-Dxd. These results indicate that 9D1-LD-38 produces more potent bystander killing than 9D1-GGFG-Dxd.

[0767] ADC drugs were co-cultured with HEK293 (CFSE-labeled) + HEK293-hClaudin18.2 mixed culture cells for 3 days, and the total amount and ratio of positive and negative cells were measured using a cell counter and flow cytometer, respectively. The results are shown in Figure 18B. The isotype control (Isotype IgG1) ADC did not have a killing effect on either positive cells HEK293-hClaudin18.2 or negative cells HEK293, whereas 9D1-LD-38 and 9D1-GGFG-Dxd had killing effects on both HEK293-hClaudin18.2 and negative cells HEK293, and the killing effect of 9D1-LD38 on negative cells HEK293 was stronger than that of 9D1-GGFG-Dxd. The above results indicate that 9D1-LD38 produces a stronger bystander killing effect than 9D1-GGFG-Dxd.

[0768] Test Example 26 (In vitro enzyme release of free toxin molecules) Preparation of Activation Buffer (10 mM cysteine ​​(ACRO, catalog number: 173600250), 15 mM EDTA, 0.1 M His-HCl, pH 5.5): Weigh 3.63 g of cysteine ​​solid, dissolve in 3 mL of 0.1 M His-HCl, pH 5.5 solution, and then add 90 μL of 0.5 M EDTA solution and mix uniformly.

[0769] Sample preparation: 1) 30 μL of activation buffer was taken, 5 μL of cathepsin B enzyme solution (0.15 mg / mL, Sino Biological, catalog number: 10483-H08H) was added, and after homogeneous mixing, the mixture was allowed to stand in a 37°C water bath for 15 minutes. 300 μL of 0.1 M His-HCl (pH 5.5) solution and 36 μL of 9D1-LD-38 (375 μg) were added, and the mixture was allowed to stand overnight in a 37°C water bath. This confirmed the enzymatic digestion of free small molecules. 2) LD-38, LD-65, and Deruxtecan (MC-GGFG-Dxd) were diluted to 14 mg / mL with DMSO. 10 μL of the diluted linker-toxin was taken, and 20 μL of activation buffer was added to each. Then, 250 μL (LD-38), 244.5 μL (LD-65), and 344.8 μL (MC-GGFG-Dxd) of activation buffer were added, and the mixtures were homogeneously mixed and left in a 37°C water bath for 15 minutes. 2.3 μL of 0.4 mg / mL cathepsin B enzyme (Sino Biological, catalog number: 10483-H08H) was taken, 950 μL of activation buffer was added, and the mixture was treated in a water bath at 37°C for 15 minutes. 230 μL of the activated linker-toxin solution was taken, and 230 μL of the activated enzyme solution (the molar ratio of linker-toxin to cathepsin B is approximately 1200:1) was added, and the mixtures were treated in a 37°C water bath for 0.5 hours, 2 hours, 4 hours, and 20 hours. At each stage of the study, 50 μL of enzyme digestion solution was collected, and 2.5 μL of 10% TFA solution was added to stop the reaction. This solution was then used to detect the enzyme cleavage kinetics, and the results are shown in Figure 19.

[0770] Preparation of negative control: 30 μL of activation buffer was taken and left in a 37°C water bath for 15 minutes. 300 μL of 0.1 M His-HCl (pH 5.5) solution and 36 μL of 9D1-LD-38 (375 μg) were added, and the mixture was treated overnight in a 37°C water bath. The parameters for RP-HPLC (Proteomix RP-1000) are shown in Table A.

[0771] [Table A]

[0772] Summary: As can be seen in Figure 19, the toxin molecules released from 9D1-LD-38 after digestion by cathepsin B enzyme coincided with the peak appearance time of the standard product, Exatecan, and no interference peak appeared in the control group (Contral) without the addition of cathepsin B enzyme. Therefore, it was determined that cathepsin B enzyme effectively cleaves the linker of 9D1-LD-38, and the cleavage product released is free Exatecan.

[0773] Table 9 shows the enzymatic cleavage kinetics of LD-38, LD-65, and Deruxtecan (MC-GGFG-Dxd). LD-38 was able to effectively release toxin molecules after cleavage by cathepsin B enzyme, and enzymatic digestion was almost complete after 4 hours (95%). LD-65 and MC-GGFG-Dxd were unresponsive to cathepsin B enzyme. After 20 hours of enzymatic digestion, only a small amount of toxin (4%) was released from LD-65, while no enzymatic cleavage product was released from MC-GGFG-Dxd.

[0774] [Table 9]

[0775] Test Example 27 (In vitro serum stability) Sample incubation: 9D1-LD-38, 9D1-LD-65, 9D1-LD-73, 9D1-LD-74, and 9D1-Deruxtecan (9D1-GGFG-Dxd), each with a DAR value of 8, prepared in the aforementioned test examples, were prepared with human, cynomolgus monkey, and rat serum (mouse serum was added to 9D1-LD-38 and 9D1-LD-65). The final concentration was 300 μg / mL, 200 μL / tube, in three overlapping wells. A negative control (0.5% BSA-PBS) and a standard were also placed simultaneously. All samples were left in a 37°C incubator for 0, 1, 3, 7, 14, and 21 days, after which the free toxin Exatecan was measured.

[0776] Sample preparation: 10.0 μL each 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 tube strip, and 25.0 μL of internal standard working solution was added (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 mixture was shaken at 2500 rpm for 5 minutes. The mixture was centrifuged for 5 minutes at 4°C and 3800 g. 80.0 μL of the supernatant was collected and dried with nitrogen at 40°C. 100 μL of 0.1% FA in 15% MeOH was added to each well, and the mixture was homogeneously mixed and subjected to LC-MS analysis.

[0777] Sample analysis: Liquid chromatography (Nexera, Shimadzu) conditions, mobile phase A: 0.1% formic acid aqueous solution, mobile phase B: 0.1% acetonitrile solution. Chromatography column: ACE Excel 3μm 300-Å, 50x2.1mm, flow rate: 0.6mL / min, column temperature: 40℃, mass spectrometry (Triple Quad 7500, AB / Sciex), data acquisition (Analyst, version 1.6.2, Applied Biosystems / Sciex).

[0778] Data processing: C 毒素The toxin content in ADC is calculated using the formula: (toxin molecular weight × DAR) / ADC molecular weight × 300 μg / mL. Serum toxin release rate = serum toxin content / C 毒素 Using Graphpad Prism9, a curve was plotted with incubation time on the x-axis and release rate on the y-axis.

[0779] Summary: Figure 20 shows the toxin Exatecan release rate-time curve in an in vitro serum stability study. Toxin release from ADC increases with incubation time. Figure 20 includes Figures 20A, 20B, 20C, 20D, 20E, and 20F. The toxin release rates of 9D1-LD-38 (Figure 20A) after incubation at 37°C for 21 days in human, cynomolgus monkey, and rat serum were only 2.8%, 0.9%, and 3.3%, respectively (the release peak in rat serum was approximately 7 days after incubation). The Dxd release rates of 9D1-Deruxtecan (9D1-GGFG-Dxd) (Figure 20B), incubated under the same conditions for 21 days, were 8.5% (human serum), 4.6% (cynomolgus monkey serum), and 5.3% (rat serum), which correspond to 3 times, 5 times, and 1.5 times the toxin release of 9D1-LD-38 in the serum of the corresponding species. The serum stability of 9D1-LD-65 (Figure 20C), 9D1-LD-73 (Figure 20D), and 9D1-LD-74 (Figure 20E) in humans, cynomolgus monkeys, and rats was essentially the same as that of 9D1-LD-38. The maximum toxin release rate in human serum was approximately 3%, while the maximum toxin release rates in cynomolgus monkey and rat serum were all less than 1.5%. 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 statistically significant, and the maximum release rates of both at day 21 did not exceed 2%.

[0780] Test Example 28 (Dynamic changes in ADC DAR values ​​in rats) Three male SD rats were selected and randomly divided into three groups. Each group received a single intravenous dose of 10 mg / kg of the test products 9D1-LD-38, 9D1-LD-65, and 9D1-Deruxtecan (9D1-GGFG-Dxd) (each with a DAR value of 8), prepared in the above-described example. Whole blood was collected from each test product group before administration, 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). The blood was divided into two parts, and serum and plasma were collected separately and stored frozen at -60°C.

[0781] Method for measuring total serum antibodies: 10.0 μL each 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 tube strip. 100 μL of precipitant (1% isopropanol solution of trichloroacetic acid) was added to each well, and the mixture was shaken at 2000 rpm for 5 minutes. The mixture was then centrifuged at 500 g at 10°C for 5 minutes, and the supernatant was removed. 100 μL of ammonium bicarbonate solution (100 mM, pH=8.0) was added to each well, and the mixture was homogeneously mixed by vortexing. The precipitate was resuspended, and the mixture was centrifuged at 1000 g for 5 minutes at 10°C, and the supernatant was removed. 50.0 μL of ammonium bicarbonate solution (100 mM, pH=8.0) was added to each well, and the mixture was homogeneously mixed by vortexing. 10.0 μL of sigmatrypsin solution (Sigma-Ace, catalog number: Y0002311) (5 mg / mL) was added to each well, centrifuged at 3500 rpm for 30 seconds, vortexed at 1000 rpm for 3 minutes, and incubated in a shaking incubator at 60°C and 550 rpm for 1.5 hours. After incubation, 20.0 μL of 10% FA in ACN (termination agent) was added to each tube. 10.0 μL of internal standard working solution (TTPP-IS, human Fc universal peptide segment, 5000 ng / ml) was added to the zero-concentration sample, standard curve sample, quality control sample, and test sample. 10.0 μL of internal standard preparation solvent (ammonium bicarbonate solution (100 mM, pH=8.0)) was added to the double blank sample. The above samples were centrifuged at 3800 g for 10 seconds and vortexed at 1200 rpm for 2 minutes. The mixture was centrifuged at 3800g for 5 minutes, 50.0 μL of the supernatant was collected, 50.0 μL of water was added to each well and mixed uniformly, and then subjected to LC-MS analysis.

[0782] Method for measuring serum-bound toxins: 10.0 μL each 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 tube strip. 90 μL of precipitant (isopropanol solution of 1% trichloroacetic acid) was added to each well, and the mixture was shaken at 2000 rpm for 5 minutes. The mixture was then 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, and the mixture was homogeneously mixed by vortexing to resuspend the precipitate. The mixture was then 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, and the mixture was homogeneously mixed by vortexing. 60.0 μL of papain (Adamas Life, catalog number: 88915D, 5 mg / mL) was added to each well, centrifuged at 3500 rpm for 30 seconds, vortexed at 1000 rpm for 3 minutes, and incubated in a shaking incubator at 40°C and 550 rpm for 2 hours. 10 μL of 1% formic acid aqueous solution was added to each well, centrifuged at 3500 rpm for 30 seconds, and incubated at 40°C and 550 rpm for 0.5 hours. 25.0 μL of internal standard working solution (Exatecan-D5, 1000 ng / ml) was added to the zero-concentration sample, standard curve sample, quality control sample, and test sample. 25.0 μL of internal standard preparation solvent was added to the double blank sample. The above samples were centrifuged at 3800 g for 10 seconds and vortexed at 1200 rpm for 2 minutes. 200 μL of pre-cooled acetonitrile was added to each well, and the mixture was shaken at 2500 rpm for 5 minutes. The mixture was then centrifuged at 3800 g for 5 minutes, and 50.0 μL of the supernatant was collected. 150 μL of 0.5% formic acid aqueous solution was added to each well, and the mixture was homogeneously mixed before being subjected to LC-MS analysis.

[0783] The total antibody (Ab) and conjugated toxin concentrations of each test product in serum samples were analyzed using LC / MS, and the conjugated anti-DAR value in animals at each time point was calculated using the formula DAR = conjugated toxin concentration (mmol / L) / total antibody concentration (mmol / L).

[0784] Summary: Figure 21 shows the DAR value-time curves of ADCs in animal serum for each test sample group. When rats were given a single intravenous injection of 10 mg / kg, the DAR values ​​of 9D1-LD-38, 9D1-LD-65, and 9D1-Deruxtecan (9D1-GGFG-Dxd) gradually decreased over time. Among these, the DAR values ​​of 9D1-LD-65 and 9D1-LD-38 showed nearly identical curves and were able to maintain relatively high DAR values. The DAR value remained at approximately 7 at 48 hours post-administration and at approximately 3-4 on day 21. The DAR value of 9D1-Deruxtecan (9D1-GGFG-Dxd) rapidly decreased from approximately 7.5 at the start of administration to 5.5 at 48 hours post-administration, and further to approximately 1.6 on day 21. 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 indicated by higher DAR.

[0785] Test Example 29 (Tissue Distribution in the Body of Tumor-Bearing Mice) Modeling and administration of tumor-bearing mice: MC38-hClaudin18.2 cells (1 × 10⁶) were introduced into the ventral region of male C57BL / 6 mice (6 weeks old). 6 (100 μL PBS) was subcutaneously inoculated. The tumors in the mice were approximately 300 mm. 3 Once the mice reached a certain volume, they were divided into groups and administered drugs. 9D1-LD-38, 9D1-LD-65, 9D1-Deruxtecan (9D1-GGFG-Dxd), and PBS, prepared in the above-described test example, were administered by tail vein injection. The DAR value for each was 8, and the dose was 10 mg / kg.

[0786] Sample collection: Blood and tumor tissue were collected 24, 72, and 168 hours after administration, with samples taken from three mice at each time point. Whole blood was collected from the orbits of tumor-bearing mice, placed in an anticoagulant tube containing EDTA-K2 anticoagulant, and centrifuged within 30 minutes to obtain plasma. After the mice were sacrificed by CO2 asphyxiation, the tumors were collected. Plasma and tumor tissue were stored at -80°C.

[0787] Measurement of free toxins in plasma: 10.0 μL each 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 tube strip, and 25.0 μL of internal standard working solution (Exatecan D5 concentration was 3.00 ng / mL; 25.0 μL of internal standard diluent was added to the double blank sample) was added. 90.0 μL of methanol:acetonitrile (1:1, v / v) was added to each well, and the mixture was shaken at 2500 rpm for 5 minutes. The mixture was centrifuged for 5 minutes at 4°C and 3800 g. 80.0 μL of the 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 homogeneously, and subjected to LC-MS analysis.

[0788] Measurement of total toxins (bound toxins + free toxins) in blood: 10.0 μL each 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 tube strips. 10.0 μL of papain (Adamas Life, catalog number: 88915D, 5 mg / ml) was added to each well, and the samples were centrifuged at 3500 rpm for 1 minute. The samples were then placed in a shaking incubator and incubated at 40°C, 550 rpm for 6-8 hours. 1.00 μL of 1% FA in H2O was added to each well, and the samples were centrifuged at 3500 rpm for 1 minute, followed by incubation at 40°C, 550 rpm for 30 minutes. 10.0 μL of internal standard working solution (Exatecan-D5, 1000 ng / ml) was added to the zero-concentration sample, standard curve sample, quality control sample, and test sample. 10.0 μL of internal standard preparation solvent was added to the double blank sample. The above sample was centrifuged at 3800 g for 10 seconds and vortexed at 1200 rpm for 2 minutes. 100 μL of pre-cooled ACN was added to each well, and the mixture was shaken at 2500 rpm for 5 minutes, followed by centrifugation at 3500 g for 5 minutes. 30.0 μL of the supernatant was collected, and 90.0 μL of 0.5% FA in H2O was added to each well, vortexed for 3 minutes, and subjected to LC-MS analysis.

[0789] Measurement of free toxins in tumor tissue: Homogenization of tumor tissue (dosage group and blank PBS group): Tumor tissue was weighed and homogenized by adding 10 times the volume of 50% methanol water. The homogenized material from the blank PBS group was used as the matrix to prepare standard curves and quality control samples. 10.0 μL each of the 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 tube strips, and 25.0 μL of internal standard working solution was added (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 mixture was shaken at 2500 rpm for 5 minutes, followed by centrifugation at 4°C and 3800 g for 5 minutes. 80.0 μL of the supernatant was collected and dried with nitrogen at 40°C. 100 μL of 0.1% FA in 15% MeOH was added to each well, vortexed for 3 minutes, and then subjected to LC-MS analysis.

[0790] Measurement of total toxins (bound toxins + free toxins) in tumor tissue: 10.0 μL each 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 tube strips. 10.0 μL of papain (Adamas Life, catalog number: 88915D 5 mg / ml) was added to each well, centrifuged at 3500 rpm for 1 minute, and incubated in a shaking incubator (40°C, 550 rpm) for 6-8 hours. 1.00 μL of 1% FA in H2O was added to each well, centrifuged at 3500 rpm for 1 minute, and incubated at 40°C, 550 rpm for 30 minutes. 5. 10.0 μL of internal standard working solution (Exatecan-D5, 1000 ng / ml) was added to the zero-concentration sample, standard curve sample, quality control sample, and test sample. 10.0 μL of internal standard preparation solvent was added to the double blank sample. The above sample was centrifuged at 3800 g for 10 seconds and vortexed at 1200 rpm for 2 minutes. 100 μL of pre-cooled ACN was added to each well and shaken at 2500 rpm for 5 minutes. Centrifuged at 3500 g for 5 minutes. 30.0 μL of the supernatant was collected, 90.0 μL of 0.5% FA in H2O was added to each well, vortexed for 3 minutes, and subjected to LC-MS analysis.

[0791] Sample analysis: Liquid chromatography (Nexera, Shimadzu) conditions: Mobile phase A: 0.1% formic acid aqueous solution, Mobile phase B: 0.1% acetonitrile solution, Chromatography column: ACE Excel 3μm 300-Å, 50x2.1mm, Flow rate: 0.6mL / min, Column temperature: 40℃, Mass spectrometry (Triple Quad 7500, AB / Sciex), Data acquisition (Analyst, version 1.6.2, Applied Biosystems / Sciex). Using GraphPad Prism9 software, sample collection time and corresponding toxin content were entered, and toxin content curves in plasma and tumor tissue were plotted. The results are shown in Figures 22 and 23. Figure 22 includes Figures 22A and 22B, where Figure 22A shows the total toxin content (bound toxins + free toxins) 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, where Figure 23A shows the total toxins (bound toxins + free toxins) in the tissues of tumor-bearing mice, and Figure 23B shows the content of free toxins in the tissues of tumor-bearing mice.

[0792] Summary: As can be seen from Figure 22, the free toxins and total toxins in the plasma of animals in each test sample group showed essentially the same trend of change, reaching peak levels at 24 hours and then slowly decreasing. Among these, the total toxin content of the 9D1-LD-38 group and the 9D1-LD-65 group (Figure 22A) were essentially identical, reaching 3000-4000 ng / mL after 24 hours and decreasing by approximately 70-80% after 7 days. The total toxin content in the plasma of 9D1-Deruxtecan (9D1-GGFG-Dxd) at each time point was only about half (1 / 2) that of 9D1-LD-38 and 9D1-LD-65. The free toxin content and trends in change in the plasma of animals in the 9D1-LD-38, 9D1-LD-65, and 9D1-Deruxtecan (9D1-GGFG-Dxd) groups were also largely consistent (Figure 22B), reaching peak levels at 24 hours and then slowly decreasing. Compared to the total toxin content in plasma, the free toxin content was extremely low, with all levels below 0.4 ng / mL after 24 hours, accounting for 0.01% to 0.02% of the total toxin content, and far below the IC50 value for in vitro target cell killing (3 ng / mL). This indicates that the toxins of the three ADC drugs are not easily eliminated in plasma and are relatively stable.

[0793] As can be seen in Figure 23, the total toxin levels in the tumor tissue of each treatment group were highest 24 hours after administration (Figure 23A). Subsequently, they rapidly decreased to about one-third of the 24-hour level by 168 hours. Among these, the concentration levels of 9D1-LD38 and 9D1-LD-65 in the tumor tissue were significantly higher than those of 9D1-Deruxtecan (9D1-GGFG-Dxd). The free toxin levels in the tumor tissue of each treatment group were highest 24 hours after administration (Figure 23B). Subsequently, they gradually decreased over 72 hours and remained nearly constant until 168 hours. The free toxin levels in the tumor tissue of 9D1-LD-38 and 9D1-LD-65 were significantly higher than those of 9D1-Deruxtecan (9D1-GGFG-Dxd) (2-5 times) throughout the entire administration period, with 9D1-LD-38 showing the best results. Based on the results regarding the total toxin (bound toxin + free toxin) and free toxin content in the tumor tissue of the above-mentioned tumor-bearing mice, it was found that 9D1-LD-38 and 9D1-LD-65 exhibit superior penetration and concentration into tumor tissue, releasing higher levels of free toxin, and thus producing superior tumor therapeutic effects compared to 9D1-Deruxtecan (9D1-GGFG-Dxd).

[0794] Test Example 30 (Pharmacological efficacy in a mouse model of p-gp-mediated drug-resistant tumors) Modeling and administration of tumor-bearing mice: MC38hClaudin18.2 cells (1 × 10⁶) were placed ventrally in BALB / c nude mice (female, 5-6 weeks old). 6 Subcutaneous inoculation was performed using PBS (100 μL / 100 mg). The tumor was approximately 120 mm. 3 Once the tumors reached a certain volume, they were randomly divided into groups according to Table 10 and administered accordingly. Tumor weight and body weight were measured twice a week, and tumor volume (TV, mm) was measured. 3 The TV (mm²) was calculated and the tumor growth curve was plotted. 3 )=(L×W 2) / 2, where L represents the tumor length and W represents the tumor width. The tumor growth curves and body weight curves of mice were plotted in XY mode using GraphPad Prism software (GraphPad Software, San Diego, California). Tumor volume and body weight are expressed as mean ± standard error. The results are shown in Figure 24, which includes Figures 24A and 24B, where Figure 24A is the mouse tumor growth curve and Figure 24B is the mouse body weight curve.

[0795] [Table 10]

[0796] Summary: P-glycoprotein (P-gp) membrane transporters cause chemotherapy resistance through active drug efflux, and chemotherapy resistance is one of the main causes of treatment failure. The mouse MC38 colorectal cancer model overexpresses p-gp and is unresponsive to many chemotherapy drugs. Figure 24A shows the tumor therapeutic effects of ADC drugs alone or in combination with a p-gp inhibitor and thaliquidal in the MC38-hClaudin18.2 tumor-bearing mouse model. With thaliquidal alone, no tumor inhibitory effect was observed compared to the PBS control group. Tumors in the 9D1-vc-PAB-MMAE group grew rapidly, and the tumor growth curve showed a weak inhibitory effect compared to the PBS control group (tumor volume at 17 days post-administration was approximately 2600 mm² in each group). 3 and 1900mm 3 ). 9D1-vc-PAB-MMAE + Taliquidal significantly inhibited tumor growth and reduced tumor size to almost complete disappearance. 9D1-Deruxtecan (9D1-GGFG-Dxd) significantly inhibited tumor growth, and the tumor inhibitory effect of 9D1-Deruxtecan (9D1-GGFG-Dxd) + Taliquidal was slightly enhanced compared to 9D1-Deruxtecan (9D1-GGFG-Dxd) alone, but neither resulted in tumor reduction (tumor volume 17 days post-administration was approximately 1000 mm² in each case). 3 , and 660mm 3). Both 9D1-LD-38 alone and 9D1-LD-38 + talikidal strongly inhibited tumor growth and reduced tumor size to almost complete disappearance. After administration, the mice's body weight remained within a normal range, and no weight loss was observed (Figure 24B). These results indicate that 9D1-LD-38 is resistant to drug resistance in tumors caused by p-gp compared to 9D1-vc-PAB-MMAE and 9D1-Deruxtecan (9D1-GGFG-Dxd), and that good tumor therapeutic effects can be obtained with monotherapy.

[0797] Test Example 31 (Anticumon effect of concomitant administration of ICI in tumor-bearing mice) Modeling and administration of tumor-bearing mice: 4T1-hClaudin18.2 cells were cultured in a rectangular flask using 1640 complete medium (10% FBS, supplemented with 2 μg / mL puromycin, Gibco, catalog number: A1113803) and placed in a 37°C, 5% CO2 incubator. When cell confluence reached approximately 80%, the cells were digested with TRYPLE EXPRESS trypsin (Gibco, catalog number: 12605028), centrifuged, and extracted to 2.5 × 10⁶ cells. 7 Individual cells were collected, washed once with PBS, the supernatant was discarded, and the cells were resuspended in PBS containing 50% Matrigel (Japanese name: Matrigel, Corning, catalog number: 326234) to a total volume of 5 ml. 100 μL / mice of the cell suspension were inoculated into the skin of the right abdomen of BALB / c female mice (female, 5-6 weeks old). The tumor was approximately 150 mm. 3 Once the tumors reached a certain volume, they were randomly divided into groups according to Table 11 and administered. Tumor size was measured twice a week with calipers. TV(mm 3 )=(L×W 2 ) / 2, where L represents the tumor length and W represents the tumor width. The tumor growth curves of mice were plotted in XY mode using GraphPad Prism software (GraphPad Software, San Diego, California), and the tumor volume was expressed as mean ± standard error.

[0798] [Table 11]

[0799] Summary: Immune checkpoint inhibitors (ICIs) are one of the foundations of cancer treatment. Figure 25 shows the combined therapeutic effect of 9D1-LD-38 and the anti-mouse PD-1 drug αPD-1 in a 4T1-hClaudin18.2 tumor-bearing mouse model of immune-normal BALB / c mice. Compared to the PBS control group, both 9D1-LD38 monotherapy and αPD-1 monotherapy showed partial tumor growth inhibition (tumor volume was approximately 850 mm² at 14 days post-administration). 3 , 480mm 3 , 426mm 3 The combination therapy of 9D1-LD38 + αPD-1 significantly inhibited tumor growth and also showed a remarkable tumor regression effect (tumor volume was approximately 70 mm on day 10 after administration). 3 ).

[0800] Test Example 32 (Antitumor effect of bystander-killing action on tumor-bearing mice) Tumor-bearing mouse modeling and administration: MC38-hClaudin18.2 cells and MC38-luc (luciferase) were cultured in DMEM medium (Meilunbio, catalog number: MA0212-Nov-02H) containing inactivated 10% fetal bovine serum (Excell Bio, catalog number: 12A222) and 1x penicillin / streptomycin (Solarbio, catalog number: 20221231) in a 37°C, 5% CO2 incubator. Tumor cells in the logarithmic growth phase were used for in vivo tumor inoculation. MC38-hClaudin18.2 cells were mixed with MC38-luc cells in a (1:1) ratio, resuspended in PBS, and the cell concentration was increased to 5 × 10⁶. 6 The solution was adjusted to 100 μL / ml and subcutaneously inoculated into the right flank of BALB / c nude mice at a dose of 100 μL / mice. The tumor was approximately 120 mm. 3 Once they reached the required volume, they were divided into four groups of five and administered (recorded as PG-D0 on this day). The specific administration regimen is as shown in the table below.

[0801] [Table 12]

[0802] The tumor volume was measured 2-3 times per week using calipers, and the longest and shortest diameters of the tumor were recorded. The volume was calculated using the following formula: Volume (TV) = 0.5 × Longest diameter × Shortest diameter × Shortest diameter. Luciferin substrate (15 mg / ml, 10 μl / g) was administered intraperitoneally to mice, and they were anesthetized with isoflurane. Luminescence signals were collected using a small animal in vivo imaging system (IVIS Lumina Series III, PerkinElmer).

[0803] Summary: Bystander killing refers to the ability of ADC drugs to release toxins after being taken up by target-positive tumor cells, and these toxins kill adjacent target-negative tumor cells via transmembrane action. This is considered one of the reasons why ADCs can exert superior efficacy against target-expressing heterogeneous tumors. In this study, a target-expressing heterogeneous tumor-bearing mouse model was created by mixing target-positive cells (MC38-hClaudin18.2) and target-negative cells (MC38-luc) and inoculating mice, and the bystander killing effect of the ADC drugs tested was verified. The results are shown in Figure 26, which includes Figures 26A and 26B. Figure 26A shows the change in tumor volume in mice after administration, and Figure 26B shows the bioluminescence signal diagrams of target-negative tumor cells in tumor-bearing mice on days 3 and 7 after administration. As can be seen from Figure 26A, compared to the PBS control group (Contral), ISO(Isotype)-LD-38 grew faster and showed only weak tumor inhibitory activity (16 days after tumor inoculation, the tumor volume was approximately 2700 mm² in each case). 3 and 1700mm 3 ). 9D1-Deruxtecan (9D1-GGFG-Dxd) was able to partially inhibit tumor growth (16 days after tumor inoculation, tumor volume was approximately 1000 mm²). 3), the tumor volume continued to increase. 9D1-LD-38 significantly inhibited tumor growth, and the tumor was almost completely removed by day 16. This indicates that 9D1-LD-38 can kill not only target-positive MC38-hClaudin18.2 cells but also target-negative MC38-luc cells. Bioluminescence signals labeled with target-negative tumor cells from tumor-bearing mice collected on day 3 (PG-3, day 3 after inoculation) and day 7 (PG-7, day 12 after inoculation) (Figure 26B) also clearly showed that 9D1-LD38 effectively kills target-negative tumor cells and significantly weakens or eliminates bioluminescence signals compared to the control group and the ISO-LD38 and 9D1-Deruxtecan (9D1-GGFG-Dxd) administered groups.

[0804] Although embodiments of the present application have been described above with reference to the drawings, the present application is not limited to the specific embodiments and applications described above, and the specific embodiments described above are merely general and indicative, not limiting. Those skilled in the art can create many more embodiments under the disclosure of this specification and without departing from the scope of protection of the claims of the present application, all of which are covered by the present application.

Claims

1. The compound represented by formula (1), whose structure is shown below, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates. 【Chemistry 1】 (M is a linking group, Sp is -L ａ -Q ａ - and A is selected from none, hydrogen, and alkyl. T is linear or branched (C １ ~C ８ ) shows a trivalent alkyl group, Y, Y', and Y'' are -L respectively ｂ -Q ｂ - and L ａ , L ｂ Each of these is selected from none, alkylene, alkenylene, alkylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, heteroarylene, or polyethylene glycol group, where the alkylene, alkenylene, alkylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, heteroarylene, and polyethylene glycol group are optionally substituted with substituents. Q ａ , Q ｂ is each independently none, -C(=O)NR-, -C(=O)O-, -OC(=O)NR-, -S(=O) ２ NR-, -NRC(=O)-, -OC(=O)-, -NRC(=O)O-, -NR S(=O) ２ -, -NRC(=O)NR-, -NHC-(=NH)NH-, -C(=O)-, -OC(=O)O-, -O-, -NR-, -S-, S(=O) ２ -, S(=O)-, or -Se-, where R is selected from hydrogen or optionally substituted alkyl, Z is selected from none, hydrogen, or optionally substituted alkyl or sulfonic acid internal salts. Z' and Z'' are selected from none, hydrogen, optionally substituted alkyl, sulfonic acid end salts, or phosphate end salts. At least one of Z, Z', and Z'' is an end salt of a sulfonic acid or an end salt of a phosphate. X includes self-destructing units or none, D contains 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 between 0 and 5.

2. The compound according to claim 1, wherein formula (1) is formula (1-1), or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof. 【Chemistry 2】 (Here, the definitions of M, Sp, A, T, Y, h, o, i, X, D, and Z are as defined in claim 1.) At least one of the i Zs is an internal sulfonic acid salt.

3. The compound according to claim 2, wherein formula (1-1) is formula (1-1-1), or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates. 【Transformation 3】 (Here, AA is an amino acid residue, and W is an integer greater than or equal to 1.)

4. The compound according to claim 1, wherein formula (1) is formula (1-2), or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates. 【Chemistry 4】 (Here, the definitions of M, Sp, A, T, Y, Z, o, p, i, h, X, Y', and D are as in claim 1, and Z' is an endosodium sulfonic acid or endosodium phosphate.)

5. The compound according to claim 4, wherein formula (1) is formula (1-2-1), or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates. 【Transformation 5】 (Here, AA is an amino acid residue, and W is an integer greater than or equal to 1.)

6. The compound according to claim 1, wherein formula (1) is formula (1-3), or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof. 【Transformation 6】 (Here, the definitions of M, Sp, A, Y, T, Z, o, i, h, q, X, Y'', and D are as in Claim 1, and Z'' is an end salt of sulfonic acid or an end salt of phosphate.)

7. The compound according to claim 6, wherein formula (1) is formula (1-3-1), or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates. 【Transformation 7】 (Here, AA is an amino acid residue, and W is an integer greater than or equal to 1.)

8. If both A and Z are absent, then N, Y, and T form a ring, i.e. 【Transformation 8】 Preferably 【Chemistry 9】 The compound according to any one of claims 1 to 7, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

9. The aforementioned amino acid is one or more selected from alanine, arginine, asparagine, aspartic acid, γ-carboxyglutamic acid, citrulline, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, n-leucine, norvaline, ornithine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine, and is a compound according to any one of claims 3, 5, or 7, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

10. The compound according to any one of claims 3, 5, or 7, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt thereof, prodrug, or solvate thereof, wherein the amino acid is one or more selected from valine, glutamic acid, citrulline, lysine, alanine, phenylalanine, arginine, glutamine, asparagine, and glycine.

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

12. M is N ３ SH, ONH ２ NH ２ CO ２ H, (alkyl)-CO-,HCO-,BrCH ２－ ICH ２ - 【Chemistry 10】 A compound according to any one of claims 1 to 11, selected from, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

13. M is, 【Chemistry 11】 The compound according to any one of claims 1 to 12, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

14. M is 【Chemistry 12】 The compound according to any one of claims 1 to 13, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

15. A is hydrogen, methyl, ethyl, or propyl, and is a compound according to any one of claims 1 to 14, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

16. A is hydrogen, and the compound according to any one of claims 1 to 15, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

17. The T is one of the following structures, and is a compound according to any one of claims 1 to 16, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof. 【Chemistry 13】

18. The aforementioned T is 【Chemistry 14】 The compound according to any one of claims 1 to 17, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

19. L ａ The compound according to any one of claims 1 to 18, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein the alkylene, alkenylene, alkynylene, saturated or unsaturated aliphatic cyclylene having 3 to 10 ring carbon atoms, saturated or unsaturated aliphatic heterocyclylene having 3 to 10 ring carbon atoms and 1 to 4 heteroatoms, arylene having 6 to 18 carbon atoms with one or more rings, heteroarylene having 6 to 18 carbon atoms and 1 to 4 heteroatoms, or polyethylene glycol group, wherein the alkylene, alkenylene, alkynylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, or heteroarylene is optionally substituted by substituents, and the heteroatoms are selected from nitrogen, oxygen, or sulfur.

20. L ａ C １ ~C １０ Linear or branched alkylenes, C ２ ~C １２ Linear or branched alkenylenes, C ２ ~C １２ One, two, or three or more selected from linear or branched alkylene, saturated or unsaturated aliphatic cyclylene having 3 to 6 ring carbon atoms, saturated or unsaturated aliphatic heterocyclylene containing a heteroatom and having 3 to 6 ring carbon atoms and having 1 to 4 heteroatoms, arylene having 6 to 10 carbon atoms having one or more rings, heteroarylene having 6 to 10 carbon atoms and having 1 to 4 heteroatoms, or polyethylene glycol group, wherein the alkylene, alkenylene, alkylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, and heteroarylene are optionally substituted with substituents, and the heteroatoms are selected from nitrogen, oxygen, or sulfur, as described in any one of claims 1 to 19, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

21. L ａ C １ ~C ６ Linear or branched alkylenes, C ２ ~C ６ Linear or branched alkenylenes, C ２ ~C ６ A compound according to any one of claims 1 to 20, comprising one, two, or three or more selected linear or branched alkylenes or polyethylene glycol groups, wherein the alkylenes, alkenylenes, or alkylenes are optionally substituted with substituents, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

22. L ａ C １ ~C ６ It consists of one, two, or three or more linear or branched alkylenes or polyethylene glycol groups, wherein the polyethylene glycol groups are -(CH ２ CH ２ O) ｎ -, -O(CH ２ CH ２ O) ｎ -, - (CH ２ CH ２ O) ｎ CH ２ -, - (CH ２ CH ２ O) ｎ CH ２ CH ２ -ien-CH ２ O(CH ２ CH ２ O) ｎ -, or -CH ２ O(CH ２ CH ２ O) ｎ CH ２ A compound according to any one of claims 1 to 21, or a tautomer, meso compound, racemic compound, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein n is an integer of 1 or more.

23. L ａ This includes ethylene, linear propylene, linear butylene, linear pentylene, or -(CH ２ CH ２ O) ｎ CH ２ CH ２ A compound according to any one of claims 1 to 22, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein n is one, two, or three or more of the following, where n is 2, 3, 4, 5, 6, 7, or 8.

24. L ａ is ethylene and / or -(CH ２ CH ２ O) ４ CH ２ CH ２ - or pentylene, the compound according to any one of claims 1 to 23, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

25. L ｂ The compound according to any one of claims 1 to 24, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof. The compound is described in any one of claims 1 to 24, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt thereof. The compound is described in any one of claims 1 to 24, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt thereof.

26. L ｂ C １ ~C １０ Linear or branched alkylenes, C ２ ~C １２ Linear or branched alkenylenes, C ２ ~C １２ One, two, or three or more compounds selected from linear or branched alkylylene, saturated or unsaturated aliphatic cyclylene with 3 to 6 ring carbon atoms, saturated or unsaturated aliphatic heterocyclylene containing a heteroatom and having 3 to 6 ring carbon atoms and having 1 to 4 heteroatoms, arylene with 6 to 10 carbon atoms having one or more rings, heteroarylene with 6 to 10 carbon atoms and having 1 to 4 heteroatoms having one or more rings, or polyethylene glycol, wherein the alkylene, alkenylene, alkylylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, and heteroarylene are optionally substituted with substituents, and the heteroatoms are selected from nitrogen, oxygen, or sulfur, as described in any one of claims 1 to 25, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

27. L ｂ is selected from C １ to C ６ linear or branched alkylene, C ２ to C ６ linear or branched alkenylene, C ２ to C ６ linear or branched alkynylene, or a polyethylene glycol group, wherein the polyethylene glycol group is -(CH ２ CH ２ O) ｎ -, -O(CH ２ CH ２ O) ｎ -, -(CH ２ CH ２ O) ｎ CH ２ -, -(CH ２ CH ２ O) ｎ CH ２ CH ２ -, -CH ２ O(CH ２ CH ２ O) ｎ -, or -CH ２ O(CH ２ CH ２ O) ｎ CH ２ -, and is one, two or more than three of them, n is an integer of 1 or more, and the alkylene, alkenylene, alkynylene are optionally substituted by substituents. The compound according to any one of claims 1 to 26, or its tautomer, meso form, racemic form, enantiomer, diastereomer, in the form of a mixture thereof, or its pharmaceutically acceptable salt, prodrug, or solvate.

28. L ｂ C １ ~C ６ Linear or branched alkylenes, or C ６ ~C １０ A compound according to any one of claims 1 to 27, which is one, two, or three or more arylenes, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

29. L ｂ The compound according to any one of claims 1 to 28, wherein methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, sec-butylene, tert-butylene, or benzylene, is one, two or three or more of these compounds, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

30. Q ａ is 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-, where R is hydrogen, or optionally substituted C １ ~C ６ A compound according to any one of claims 1 to 29, selected from linear or branched alkyl groups, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

31. Q ａ is one, two or three or more of -C(=O)-, -NR-, -C(=O)NR-, -NRC(=O)NR-, -OC(=O)O-, -OC(=O)NR-, -NRC(=O)-, or -OC(=O)-, where R is hydrogen or optionally substituted C １ ~C ６ A compound according to claim 30, selected from linear or branched alkyl groups, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

32. Q ａ is one, two, or three or more of -C(=O)-, -NR-, -NRC(=O)-, and -(C=O)NR-, where R is hydrogen or optionally substituted C １ ~C ６ A compound according to claim 30, selected from linear or branched alkyl groups, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

33. Q ａ is -C(=O)-, -N(CH ３ )-, -NHC(=O)-, -C(=O)-NH-, or -C(=O)-N(CH ３ ) - the compound according to claim 30, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

34. Q ｂ is one, two or more selected from none, -C(=O)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-, where R is hydrogen, or optionally substituted C １ ~C ６ A compound according to any one of claims 1 to 33, selected from linear or branched alkyl groups, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

35. Q ｂ C is one, two or three or more of the following: none, -C(=O)-, -C(=O)NR-, -OC(=O)NR-, -NRC(=O)-, -NRC(=O)O-, or -NRC(=O)NR-, where R is hydrogen, or optionally substituted C １ ~C ６ A compound according to claim 34, selected from linear or branched alkyl groups, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

36. Q ｂ The compound according to claim 34, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof, wherein R is none, -C(=O)-, -NRC(=O)-, -NRC(=O)NR-, or -C(=O)NR-, and R is hydrogen or methyl.

37. X is a compound according to any one of claims 1 to 36, selected from the following, in the form of a tautomer, meso compound, racemic compound, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof. 【Chemistry 15】 (The wavy line on the right in X is connected to D, R ４ is hydrogen, C １ ~C ６ Linear or branched alkyl, -(CH ２ CH ２ O) ｍ R ２ ien-CH ２ CH ２ NR ２ R ３ ien-CH ２ CH ２ SO ２ R ２ Selected from, R ２ , R ３ These are hydrogen and C, respectively. １ ~C ６ (Selected from linear or branched alkyl groups, where m is an integer greater than 1.)

38. X is, 【Chemistry 16】 And, R ４ is hydrogen, methyl, -CH ２ CH ２ OCH ２ CH ２ OH, -CH ２ CH ２ NMe ２ ien-CH ２ CH ２ SO ２ The compound according to claim 37, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

39. X is, 【Chemistry 17】 And R ４ The compound according to claim 37, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein is hydrogen or methyl.

40. The sulfonic acid salt is one of the following: a compound according to any one of claims 1 to 39, or a tautomer, meso compound, racemic compound, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof. [Chemistry 18] (The wavy line is connected to Y, Y', or Y'', Here, c, d, e, f, and g are all integers of 1, 2, 3, 4, 5, or 6. R ６ , R ７ , R １１ , R １２ , R １３ , R １４ These are none, hydrogen, and C, respectively. １ ~C ６ (Selected from linear or branched alkyl groups.)

41. The sulfonic acid salt is the compound described in claim 40, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof. 【Chemistry 19】 (Here, R ６ , R ７ , R １１ , R １２ , R １３ and R １４ Each is selected from none, methyl, or ethyl. c, d, e, f, and g are 1, 2, or 3, respectively.

42. The sulfonic acid salt is one of the following structures, and is the compound according to claim 40, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof. 【Chemistry 20】

43. The phosphate salt is one of the following: a compound according to any one of claims 1 to 42, or a tautomer, meso compound, racemic compound, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof. 【Chemistry 21】 (Here, a and b are all integers of 1, 2, 3, 4, 5, or 6, and R ８ , R ９ , and R １０ These are hydrogen and C, respectively. １ ~C ６ (Selected from linear or branched alkyl groups.)

44. The phosphate salt is the compound described in claim 43, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof. 【Chemistry 22】 (Here, R ８ , R ９ , R １０ (wherein 1 is methyl or ethyl, and 'a' is 1, 2, or 3, and 'b' is 1, 2, or 3.)

45. The phosphate salt is the compound described in claim 43, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof. 【Chemistry 23】 (Here, R ８ , R ９ , R １０ (All are methyl, a is 2, and b is 1.)

46. The D is a compound according to any one of claims 1 to 45, comprising a cytotoxin or an immunoagonist, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

47. The cytotoxin is one or two selected from tubulin inhibitors, DNA synthesis inhibitors, topoisomerase inhibitors, or RNA polymerase 2 inhibitors, and is the compound according to claim 46, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

48. The cytotoxin is one or more selected from auristatins, meitansinoid DMs, calicheamycin, duocarmycin, pyrrolobenzodiazepines, camptothecin derivatives, and α-amanitin, and is the compound according to claim 46, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

49. The immunoagonist is selected from a TLR7 / 8 agonist or a STING agonist and is a compound according to claim 46, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

50. i is 2, 3, 4, 5, 6, 7, or 8, and is a compound according to any one of claims 1 to 49, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

51. The compound according to any one of claims 1 to 50, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, 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 in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein o, p, and q are 0, 1, 2, or 3, respectively.

53. L ａ C １ ~C ６ It consists of one, two, or three or more linear or branched alkylenes, or polyethylene glycol groups. The polyethylene glycol group is -(CH ２ CH ２ O) ｎ -, -O(CH ２ CH ２ O) ｎ -, - (CH ２ CH ２ O) ｎ CH ２ -, - (CH ２ CH ２ O) ｎ CH ２ CH ２ -ien-CH ２ O(CH ２ CH ２ O) ｎ -, or -CH ２ O(CH ２ CH ２ O) ｎ CH ２ - and n is an integer greater than or equal to 1, Q ａ is one, two, or three or more of -C(=O)-, -NR-, -NR(C=O)-, and -(C=O)NR-, where, R is hydrogen, C １ ~C ６ A compound according to any one of claims 1 to 52, selected from linear or branched alkyl groups, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

54. L ａ This includes ethylene, linear propylene, linear butylene, linear pentylene, or -(CH ２ CH ２ O) ｎ CH ２ CH ２ - is one, two or three or more of the following, where n is 2, 3, 4, 5, 6, 7 or 8, Q ａ The compound according to claim 53, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof, wherein R is one, two, or three or more of -C(=O)-, -NR-, -NR(C=O)-, and -(C=O)NR-, where R is methyl and / or hydrogen.

55. L ａ is ethylene and / or -(CH ２ CH ２ O) ４ CH ２ CH ２ - or pentylene, Q ａ is -C(=O)-, -N(CH ３ )-, -NH(C=O)-, -C(=O)NH-, or -C(=O)-N(CH ３ ) - The compound according to claim 53, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

56. L ｂ C １ ~C ６ Linear or branched alkylenes, C ２ ~C ６ Linear or branched alkenylenes, C ２ ~C ６ Selected from linear or branched alkynylenes, arylenes having one or more rings and 6 to 10 carbon atoms, or polyethylene glycol groups, The polyethylene glycol group is -(CH ２ CH ２ O) ｎ -, -O(CH ２ CH ２ O) ｎ -, - (CH ２ CH ２ O) ｎ CH ２ -, - (CH ２ CH ２ O) ｎ CH ２ CH ２ -ien-CH ２ O(CH ２ CH ２ O) ｎ -, or -CH ２ O(CH ２ CH ２ O) ｎ CH ２ - is one, two, or three or more of the above, n is an integer of 1 or more, and the alkylene, alkenylene, and alkynylene are optionally substituted by substituents. Q ｂ None, -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-, one, two or more selected from these, where R is hydrogen, or optionally substituted C １ ~C ６ A compound according to any one of claims 1 to 55, selected from linear or branched alkyl groups, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

57. L ｂ C １ ~C ６ Linear or branched alkylenes, or C ６ ~C １０ It is one, two, or three or more of the allerenes. Q ｂ is one, two or three or more of the following: none, -C(=O)NR-, -C(=O)-, -OC(=O)NR-, -NRC(=O)-, -NRC(=O)O-, or -NRC(=O)NR-, where R is hydrogen, C １ ~C ６ A compound according to claim 56, selected from linear or branched alkyl groups, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

58. L ｂ This is one, two, or three or more of methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, sec-butylene, tert-butylene, or bendiylene. Q ｂ The compound according to claim 56, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof, wherein R is none, -C(=O)-, -NRC(=O)-, -NRC(=O)NR-, or -C(=O)NR-, and R is hydrogen or methyl.

59. The substituent is selected from deuterium, tritium, halogen atoms, amino, hydroxy, cyano, nitro, optionally substituted alkyl, alkenyl, alkynyl groups, optionally substituted cyclyl groups, acyl, optionally substituted amino, optionally substituted carbamoyl, optionally substituted thiocarbamoyl, optionally substituted sulfamoyl, optionally substituted hydroxy, optionally substituted thioalkyl (SH), and optionally substituted silyl, and is a compound according to any one of claims 1 to 58, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

60. The compound is a compound according to any one of claims 1 to 59, selected from the following structures, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof. 【Chemistry 24】 【Chemistry 25】 【Chemistry 26】

61. The compound represented by formula (2), whose structure is shown below, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates. 【Chemistry 27】 (Ab comprises an antibody or its antigen-binding fragment, M ’ is a linking group, Sp is -L ａ -Q ａ - and A is selected from none, hydrogen, and alkyl. T is linear or branched (C １ ~C ８ ) shows a trivalent alkyl group, Y, Y', and Y'' are -L respectively ｂ -Q ｂ - and L ａ , L ｂ Each of these is selected from none, alkylene, alkenylene, alkylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, heteroarylene, or polyethylene glycol group, where the alkylene, alkenylene, alkylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, heteroarylene, and polyethylene glycol group are optionally substituted with substituents. Q ａ Q ｂ These are none, -C(=O)NR-, -C(=O)O-, -OC(=O)NR-, and -S(=O). ２ NR-, -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-, where R is selected from hydrogen or optionally substituted alkyl groups. Z is selected from none, hydrogen, or optionally substituted alkyl or sulfonic acid internal salts. Z' and Z'' are selected from none, hydrogen, optionally substituted alkyl, sulfonic acid end salts, or phosphate end salts. At least one of Z, Z', and Z'' is an end salt of a sulfonic acid or an end salt of a phosphate. X includes self-destructing units or none, D contains 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, j is an integer greater than or equal to 1.

62. The compound according to claim 61, wherein formula (2) is formula (2-1), or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates. 【Chemistry 28】 (Here, Ab, M ’ The definitions of Sp, A, T, Y, h, o, i, j, X, D, and Z are as set forth in claim 61. At least one of the i Zs is an internal sulfonic acid salt.

63. The compound according to claim 62, wherein formula (2-1) is formula (2-1-1), or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates. 【Chemistry 29】 (Here, AA is an amino acid residue, W is an integer greater than or equal to 1.

64. The compound according to claim 61, wherein formula (2) is formula (2-2), or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates. 【Transformation 30】 (Here, Ab, M ’ The definitions of Sp, A, T, Y, Z, o, p, i, h, j, X, Y', and D are as defined in claim 61, where Z' is an endosodium sulfonic acid or endosodium phosphate.

65. The compound according to claim 64, wherein formula (2-2) is formula (2-2-1), or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates. 【Chemistry 31】 (Here, AA is an amino acid residue, and W is an integer greater than or equal to 1.)

66. The compound according to claim 61, wherein formula (2) is formula (2-3), or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof. 【Chemistry 32】 (Here, Ab, M ’ The definitions of Sp, A, Y, T, Z, o, i, h, j, q, X, Y'', and D are as in claim 61, where Z'' is an end salt of sulfonic acid or an end salt of phosphate.

67. The compound according to claim 66, wherein formula (2-3) is formula (2-3-1), or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates. 【Transformation 33】 (Here, AA is an amino acid residue, and W is an integer greater than or equal to 1.)

68. If both A and Z are absent, then N, Y, and T form a ring, i.e., 【Transformation 34】 Preferably 【Chemistry 35】 The compound according to any one of claims 61 to 67, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

69. The aforementioned amino acid is one or more selected from alanine, arginine, asparagine, aspartic acid, γ-carboxyglutamic acid, citrulline, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, n-leucine, norvaline, ornithine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine, and is a compound according to any one of claims 63, 65, or 67, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

70. The compound according to any one of claims 63, 65, or 67, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein the amino acid is one or more selected from valine, glutamic acid, citrulline, lysine, alanine, phenylalanine, arginine, glutamine, asparagine, and glycine.

71. The compound according to any one of claims 63, 65, or 67, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein the amino acid is one or more selected from valine, citrulline, alanine, phenylalanine, glutamine, and glycine.

72. M ’ The compound according to any one of claims 61 to 71, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, selected from the following structures. 【Transformation 36】 (* is concatenated to Ab.)

73. M ’ The compound according to any one of claims 61 to 72, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, selected from the following structures. 【Chemistry 37】

74. M ’ teeth, 【Transformation 38】 The compound according to any one of claims 61 to 73, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

75. A is hydrogen, methyl, ethyl, or propyl, and is a compound according to any one of claims 61 to 74, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

76. A is hydrogen, and the compound according to any one of claims 61 to 75, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

77. The T is one of the following structures, in the form of a compound according to any one of claims 61 to 76, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof. 【Chemistry 39】

78. The aforementioned T is, 【Chemistry 40】 The compound according to any one of claims 61 to 77, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

79. L ａ The compound according to any one of claims 61 to 78, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof. The compound is further described by: alkylene, alkenylene, alkynylene, saturated or unsaturated aliphatic cyclylene having 3 to 10 ring carbon atoms, saturated or unsaturated aliphatic heterocyclylene having 3 to 10 ring carbon atoms and 1 to 4 heteroatoms, arylene having one or more rings and 6 to 18 carbon atoms and 1 to 4 heteroatoms, or polyethylene glycol group, wherein the alkylene, alkenylene, alkynylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, or heteroarylene is optionally substituted by substituents, and the heteroatoms are selected from nitrogen, oxygen, or sulfur.

80. L ａ C １ ~C １０ Linear or branched alkylenes, C ２ ~C １２ Linear or branched alkenylenes, C ２ ~C １２ One, two, or three or more selected from linear or branched alkylenes, saturated or unsaturated aliphatic cyclylenes having 3 to 6 ring carbon atoms, saturated or unsaturated aliphatic heterocyclylenes containing heteroatoms and having 3 to 6 ring carbon atoms and having 1 to 4 heteroatoms, arylenes having 6 to 10 carbon atoms and having one or more rings, heteroarylenes having 6 to 10 carbon atoms and having 1 to 4 heteroatoms, or polyethylene glycol groups, wherein the alkylenes, alkenylenes, alkylenes, aliphatic cyclylenes, aliphatic heterocyclylenes, arylenes, and heteroarylenes are optionally substituted with substituents, and the heteroatoms are selected from nitrogen, oxygen, or sulfur, as described in any one of claims 61 to 79, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

81. L ａ C １ ~C ６ Linear or branched alkylenes, C ２ ~C ６ Linear or branched alkenylenes, C ２ ~C ６ A compound according to any one of claims 61 to 80, comprising one, two, or three or more selected linear or branched alkylylenes or polyethylene glycol groups, wherein the alkylenes, alkenylenes, or alkylylenes are optionally substituted with substituents, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

82. L ａ C １ ~C ６ It consists of one, two, or three or more linear or branched alkylenes, or polyethylene glycol groups. The polyethylene glycol group is -(CH ２ CH ２ O) ｎ -, -O(CH ２ CH ２ O) ｎ -, - (CH ２ CH ２ O) ｎ CH ２ -, - (CH ２ CH ２ O) ｎ CH ２ CH ２ -ien-CH ２ O(CH ２ CH ２ O) ｎ -, or -CH ２ O(CH ２ CH ２ O) ｎ CH ２ A compound according to any one of claims 61 to 81, or a tautomer, meso compound, racemic compound, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein n is an integer of 1 or more.

83. L ａ This includes ethylene, linear propylene, linear butylene, linear pentylene, or -(CH ２ CH ２ O) ｎ CH ２ CH ２ A compound according to any one of claims 61 to 82, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein n is one, two, or three or more of the following, where n is 2, 3, 4, 5, 6, 7, or 8.

84. L ａ is ethylene and / or -(CH ２ CH ２ O) ４ CH ２ CH ２ - or pentylene, the compound according to any one of claims 61 to 83, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

85. L ｂ This is one, two, or three or more selected from alkylene, alkenylene, alkynylene, saturated or unsaturated aliphatic cyclylene with 3 to 10 ring carbon atoms, saturated or unsaturated aliphatic heterocyclylene with 3 to 10 ring carbon atoms and 1 to 4 heteroatoms, arylene with 6 to 18 carbon atoms having one or more rings, heteroarylene with 6 to 18 carbon atoms having one or more rings and 1 to 4 heteroatoms, or polyethylene glycol. The alkylene, alkenylene, alkynylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, heteroarylene is optionally substituted with substituents, wherein the heteroatom is selected from nitrogen, oxygen, or sulfur, and the compound according to any one of claims 61 to 84, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

86. L ｂ C １ ~C １０ Linear or branched alkylenes, C ２ ~C １２ Linear or branched alkenylenes, C ２ ~C １２ One, two, or three or more selected from linear or branched alkynylenes, saturated or unsaturated aliphatic cyclylenes with 3 to 6 ring carbon atoms, saturated or unsaturated aliphatic heterocyclylenes containing heteroatoms and having 3 to 6 ring carbon atoms and 1 to 4 heteroatoms, arylenes with 6 to 10 carbon atoms having one or more rings, heteroarylenes with 6 to 10 carbon atoms and 1 to 4 heteroatoms having one or more rings, or polyethylene glycol. The alkylene, alkenylene, alkynylene, aliphatic cyclylene, aliphatic heterocyclylene, arylene, heteroarylene is optionally substituted with substituents, wherein the heteroatom is selected from nitrogen, oxygen, or sulfur, and the compound according to any one of claims 61 to 85, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

87. L ｂ C １ ~C ６ Linear or branched alkylenes, C ２ ~C ６ Linear or branched alkenylenes, C ２ ~C ６ Selected from linear or branched alkylylenes, or polyethylene glycol groups, The polyethylene glycol group is -(CH ２ CH ２ O) ｎ -, -O(CH ２ CH ２ O) ｎ -, - (CH ２ CH ２ O) ｎ CH ２ -, - (CH ２ CH ２ O) ｎ CH ２ CH ２ -ien-CH ２ O(CH ２ CH ２ O) ｎ -, or -CH ２ O(CH ２ CH ２ O) ｎ CH ２ - is one, two, or three or more of the above, and n is an integer greater than or equal to 1. The alkylene, alkenylene, and alkynylene are optionally substituted with substituents, and the compounds according to any one of claims 61 to 86, or in the form of tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

88. L ｂ C １ ~C ６ Linear or branched alkylenes, or C ６ ~C １０ A compound according to any one of claims 61 to 87, which is one, two, or three or more arylenes, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

89. L ｂ The compound according to any one of claims 61 to 88, wherein methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, sec-butylene, tert-butylene, or benzylene, is one, two or three or more of these compounds, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

90. Q ａ is 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-, where R is C which is optionally substituted with hydrogen. １ ~C ６ A compound according to any one of claims 61 to 89, selected from linear or branched alkyl groups, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

91. Q ａ is one, two or three or more of -C(=O)-, -NR-, -C(=O)NR-, -NRC(=O)NR-, -OC(=O)O-, -OC(=O)NR-, -NRC(=O)-, or -OC(=O)-, where, R is hydrogen or optionally substituted C １ ~C ６ A compound according to claim 90, selected from linear or branched alkyl groups, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

92. Q ａ is one, two, or three or more of -C(=O)-, -NR-, -NRC(=O)-, and -(C=O)NR-, where, R is hydrogen or optionally substituted C １ ~C ６ A compound according to claim 90, selected from linear or branched alkyl groups, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

93. Q ａ is -C(=O)-, -N(CH ３ )-, -NHC(=O)-, -C(=O)-NH-, or -C(=O)-N(CH ３ ) - the compound according to claim 90, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

94. Q ｂ is one, two or three or more selected from none, -C(=O)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-, where R is hydrogen or optionally substituted C １ ~C ６ A compound according to any one of claims 61 to 93, selected from linear or branched alkyl groups, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

95. Q ｂ is one, two or three or more of the following: none, -C(=O)-, -C(=O)NR-, -OC(=O)NR-, -NRC(=O)-, -NRC(=O)O-, or -NRC(=O)NR-, where R is hydrogen or optionally substituted C １ ~C ６ A compound according to claim 94, selected from linear or branched alkyl groups, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

96. Q ｂ The compound according to claim 94, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof, wherein R is none, -C(=O)-, -NRC(=O)-, -NRC(=O)NR-, or -C(=O)NR-, and R is hydrogen or methyl.

97. X is a compound according to any one of claims 61 to 96, selected from the following: a tautomer, meso compound, racemic compound, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof. 【Chemistry 41】 (The wavy line on the right in X is connected to D, R ４ is hydrogen, C １ ~C ６ Linear or branched alkyl, -(CH ２ CH ２ O) ｍ R ２ ien-CH ２ CH ２ NR ２ R ３ ien-CH ２ CH ２ SO ２ R ２ - Selected from, R ２ , R ３ These are hydrogen and C, respectively. １ ~C ６ (Selected from linear or branched alkyl groups, where m is an integer greater than 1.)

98. X is, 【Chemistry 42】 And, R ４ is hydrogen, methyl, -CH ２ CH ２ OCH ２ CH ２ OH, -CH ２ CH ２ NMe ２ ien-CH ２ CH ２ SO ２ The compound according to claim 97, or its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.

99. X is, 【Chemistry 43】 And R ４ The compound according to claim 97, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein is hydrogen or methyl.

100. The sulfonic acid salt is one of the following: a compound according to any one of claims 61 to 99, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof. 【Chemistry 44】 (The wavy line is connected to Y, Y', or Y'', Here, c, d, e, f, and g are all integers of 1, 2, 3, 4, 5, or 6. R ６ , R ７ , R １１ , R １２ , R １３ , R １４ These are none, hydrogen, and C, respectively. １ ~C ６ (Selected from linear or branched alkyl groups.)

101. The sulfonic acid salt is the compound according to claim 100, or in the form of its tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof. 【Chemistry 45】 (Here, R ６ , R ７ , R １１ , R １２ , R １３ and R １４ Each is selected from none, methyl, or ethyl. c, d, e, f, and g are 1, 2, or 3, respectively.

102. The sulfonic acid salt is one of the following structures, and is the compound according to claim 100, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof. 【Chemistry 46】

103. The phosphate salt is one of the following: a compound according to any one of claims 61 to 102, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof. 【Chemistry 47】 (Here, a and b are all integers of 1, 2, 3, 4, 5, or 6, and R ８ , R ９ , and R １０ These are hydrogen and C, respectively. １ ~C ６ (Selected from linear or branched alkyl groups.)

104. The phosphate salt is the compound according to claim 103, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates. 【Chemistry 48】 (Here, R ８ , R ９ , R １０ (wherein 1 is methyl or ethyl, and 'a' is 1, 2, or 3, and 'b' is 1, 2, or 3.)

105. The phosphate salt is the compound according to claim 103, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates. 【Chemistry 49】 (Here, R ８ , R ９ , R １０ (All are methyl, a is 2, and b is 1.)

106. The D is a compound according to any one of claims 61 to 105, comprising a cytotoxin or an immunoagonist, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

107. The cytotoxin is one or two selected from tubulin inhibitors, DNA synthesis inhibitors, topoisomerase inhibitors, or RNA polymerase 2 inhibitors, according to claim 106, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

108. The cytotoxin is one or more selected from auristatins, meitansinoid DMs, calicheamycin, duocarmycin, pyrrolobenzodiazepines, camptothecin derivatives, and α-amanitin, and is the compound according to claim 106, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

109. The immunoagonist is selected from a TLR7 / 8 agonist or a STING agonist and is the compound according to claim 106, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

110. i is 2, 3, 4, 5, 6, 7, or 8, the compound according to any one of claims 61 to 109, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

111. The compound according to any one of claims 61 to 110, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, 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 to 111, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein o, p, and q are 0, 1, 2, or 3, respectively.

113. L ａ C １ ~C ６ It consists of one, two, or three or more linear or branched alkylenes, or polyethylene glycol groups. The polyethylene glycol group is -(CH ２ CH ２ O) ｎ -, -O(CH ２ CH ２ O) ｎ -, - (CH ２ CH ２ O) ｎ CH ２ -, - (CH ２ CH ２ O) ｎ CH ２ CH ２ -ien-CH ２ O(CH ２ CH ２ O) ｎ -, or -CH ２ O(CH ２ CH ２ O) ｎ CH ２ - and n is an integer greater than or equal to 1, Q ａ is one, two, or three or more of -C(=O)-, -NR(C=O)-, and -(C=O)NR-, where, R is hydrogen, C １ ~C ６ A compound according to any one of claims 61 to 112, selected from linear or branched alkyl groups, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

114. L ａ This includes ethylene, linear propylene, linear butylene, linear pentylene, or -(CH ２ CH ２ O) ｎ CH ２ CH ２ - is one, two, or three or more of the following, where n is 2, 3, 4, 5, 6, 7, or 8. Q ａ The compound according to any one of claims 61 to 113, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein R is one, two or three or more of -C(=O)-, -NR-, -NR(C=O)-, and -(C=O)NR-, where R is methyl and / or hydrogen.

115. L ａ is ethylene and / or -(CH ２ CH ２ O) ４ CH ２ CH ２ - or pentylene, Q ａ is -C(=O)-, -N(CH ３ )-, -NH(C=O)-, -C(=O)NH-, or -C(=O)-N(CH ３ ) - the compound according to claim 114, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates.

116. L ｂ C １ ~C ６ Linear or branched alkylenes, C ２ ~C ６ Linear or branched alkenylenes, C ２ ~C ６ Selected from linear or branched alkynylenes, arylenes having one or more rings and 6 to 10 carbon atoms, or polyethylene glycol groups, The polyethylene glycol group is -(CH ２ CH ２ O) ｎ -, -O(CH ２ CH ２ O) ｎ -, - (CH ２ CH ２ O) ｎ CH ２ -, - (CH ２ CH ２ O) ｎ CH ２ CH ２ -ien-CH ２ O(CH ２ CH ２ O) ｎ -, or -CH ２ O(CH ２ CH ２ O) ｎ CH ２ - is one, two, or three or more of the above, n is an integer of 1 or more, and the alkylene, alkenylene, and alkynylene are optionally substituted by substituents. Q ｂ None, -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-, one, two or more selected from these, where R is hydrogen, or optionally substituted C １ ~C ６ A compound according to any one of claims 61 to 115, selected from linear or branched alkyl groups, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

117. L ｂ C １ ~C ６ Linear or branched alkylenes, or C ６ ~C １０ It is one, two, or three or more of the allerenes. Q ｂ C is one, two or three or more of the following: none, -C(=O)-, -C(=O)NR-, -OC(=O)NR-, -NRC(=O)-, -NRC(=O)O-, or -NRC(=O)NR-, where R is hydrogen, or optionally substituted C １ ~C ６ A compound according to claim 116, selected from linear or branched alkyl groups, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

118. L ｂ This is one, two, or three or more of methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, sec-butylene, tert-butylene, or bendiylene. Q ｂ The compound according to claim 116, or in the form of its tautomers, meso compounds, racemic compounds, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof, wherein R is none, -C(=O)-, -NRC(=O)-, -NRC(=O)NR-, or -C(=O)NR-, and R is hydrogen or methyl.

119. The substituent is selected from deuterium, tritium, halogen atoms, amino, hydroxy, cyano, nitro, optionally substituted alkyl, alkenyl, alkynyl groups, optionally substituted cyclyl groups, acyl, optionally substituted amino, optionally substituted carbamoyl, optionally substituted thiocarbamoyl, optionally substituted sulfamoyl, optionally substituted hydroxy, optionally substituted thioalkyl (SH), and optionally substituted silyl, and is in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

120. j is an integer from 1 to 20, and the compound according to any one of claims 61 to 119, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

121. The compound is a compound according to any one of claims 61 to 120, selected from the following structures, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof. [Transformation 50] 【Chemistry 51】 【Chemistry 52】

122. The antibody is selected from the group consisting of mouse antibodies, chimeric antibodies, humanized antibodies, and fully human antibodies, and is a compound according to any one of claims 61 to 121, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

123. The antibody is a compound according to any one of claims 61 to 122, including a monoclonal antibody, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

124. The antibody is a compound according to any one of claims 61 to 123, including a bispecific antibody, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

125. The antigen-binding fragment is Fab, Fab ’ , Fv fragment, F(ab ’ ) ２ F(ab) ２ A compound according to any one of claims 61 to 124, selected from the group consisting of scFv, di-scFv, and VHH, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

126. Preface Ab is, anti-Crowden 18.2 antibody, anti-folic acid 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-TM-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 Anti-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-STEP antibody, anti-SLITRK6 antibody, anti-ETBR antibody, anti-BCMA antibody, anti-CEACAM5 antibody, anti-SC-16 antibody, anti-SLC39A6 antibody, anti-Delta-like antibody Protein 3 antibody, anti-Claudin 18.2 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-LewisY 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-LA A compound according to any one of claims 61 to 125, selected from the group consisting of MP-1 antibody, anti-LRRC15 antibody, anti-Ly6E antibody, anti-MFI2 antibody, anti-NOTCH3 antibody, anti-p-cadherin antibody, anti-PRLR antibody, anti-RNF43 antibody, and antigen-binding fragments of the above antibodies, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

127. The heavy chains HCDR1, HCDR2, and HCDR3 of Ab and the light chains LCDR1, LCDR2, and LCDR3 are, respectively, the compound according to any one of claims 61 to 126, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

128. The heavy chain variable region VH and light chain variable region VL of Ab each comprise the heavy chain variable region VH and light chain variable region VL of an antibody, respectively, and the compound according to any one of claims 61 to 127, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

129. The heavy chain and light chain of Ab are, respectively, the compound according to any one of claims 61 to 128, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

130. The compound according to any one of claims 61 to 129, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein Ab is an anti-claudin 18.2 antibody or its antigen-binding fragment.

131. The compound according to any one of claims 61 to 130, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein Ab is an anti-human folate receptor antibody or an antigen-binding fragment thereof.

132. A pharmaceutical composition comprising a compound according to any one of claims 1 to 131, or a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

133. Use of a compound according to any one of claims 1 to 132, or in the form of a tautomer, meso, racemic, enantiomer, diastereomer, mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, in the preparation of a drug for treating and / or preventing tumors.

134. The aforementioned tumor contains 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 cycle 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 The use according to claim 133, selected from tumors associated with the expression of targets from the group consisting of 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 the tumor associated with the expression of the target includes tumors in which the target is highly expressed and / or tumors in which the target is positive.

136. The use according to claim 133, wherein the tumor is selected from solid tumors, hematological malignancies, 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, digestive tract cancer, pancreatic cancer, thyroid cancer, colorectal cancer, kidney cancer, lung cancer (e.g., non-small cell lung cancer), liver cancer, stomach 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.

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