Ligand-drug conjugates containing hydrophilic sugar structures

Abstract

A ligand drug conjugate comprising a hydrophilic sugar structure (SU), a pharma- ceutically acceptable salt or solvate thereof, a process for preparing the ligand drug conjugate, and its use in the preparation of a medicament for treating a disease, the ligand drug conjugate having improved hydrophilicity.

Description

[Technical field]

[0001] The present invention discloses ligand-drug conjugates with improved hydrophilicity, as well as methods for their preparation and their use. Specifically, the present invention discloses ligand-drug conjugates comprising hydrophilic sugar structures (SU), methods for their preparation and their use in the preparation of drugs for treating diseases. [Background technology]

[0002] Ligand-drug conjugates, especially antibody-drug conjugates, are formed by binding a targeting ligand to a small molecule drug (cytotoxin). Antibody-drug conjugates not only have the strong lethality of cytotoxic drugs, but also have the high targeting, stability and good pharmacokinetic properties of the ligands, and mainly exhibit the following characteristics: high therapeutic effect; high specificity; weak immunogenicity and low drug resistance; long circulation time in serum; and weak toxicity to non-target sites. Currently, there are more than 100 antibody-drug conjugates in clinical trials, most of which have progressed from Phase I to Phase II. Phase III trials of some antibody-drug conjugates have shown favorable results. As of November 2021, 14 types of ADCs (antibody-drug conjugates) have been launched worldwide, with their therapeutic areas being mainly hematological tumors and solid tumors, and are mainly used for second-line treatment of patients, including those with terminal, relapsed / refractory and metastatic tumor indications. A common challenge in the design of ligand-drug conjugates is the hydrophobicity of the payload, which may cause problems with water solubility, aggregation, and rapid clearance of the conjugate. Studies have shown that the amplification of the MDR1 gene and overexpression of its product (energy-dependent transporter P protein) in cancer cells may pump anticancer drugs out of the tumor cells, reducing the intracellular drug concentration and weakening or eliminating the cytotoxic effect of hydrophobic anticancer drugs, which is the main reason why tumor cells acquire multidrug resistance. Currently, the main way to overcome this problem is to improve the aqueous solubility of the ligand-drug conjugates using a polyethylene glycol (PEG) linker (see patent application WO2015057699A2). The marketed sacituzumab-govitecan (IMMU-132) solves this hydrophobic problem by attaching a cleavable maleimide linker with a short PEGylation unit to SN-38. Summary of the Invention

[0003] To solve the hydrophobicity problem of payload in ligand drug conjugates, the present invention discloses a ligand drug conjugate comprising a hydrophilic sugar structure. The present invention further discloses a linker drug conjugate comprising a hydrophilic sugar structure, and the use of the ligand drug conjugate or the linker drug conjugate in the preparation of a drug for treating or preventing a disease (e.g., a tumor or an autoimmune disease). Specifically, by linking a sugar structure (e.g., monosaccharide, disaccharide, or polysaccharide) to the payload, the hydrophilicity of the sugar is utilized to reduce the hydrophilicity of the payload, thereby improving the hydrophilicity of the ligand-drug conjugate. Compared with the prior art, the hydrophilic structures involved in the present invention are more readily available sugars, and it is easier to achieve coupling of the payload with the naturally occurring reactive sites in the sugar. Furthermore, tumor cells have a much higher ability to take up sugars than normal cells, further avoiding the non-target side effects of the ligand-drug conjugate.

[0004] In one aspect, the present invention provides a compound of formula I, II or III: [ka] In L is selected from a ligand; M is selected from any linker unit or a bond; A is selected from any linking scaffold; B is present or absent and, if present, is selected from any linking structure or bond; C is present or absent and, when present, is selected from any branching unit or bond; D, D 1 , D 2 are the same or different and are each independently selected from drugs; SU is selected from sugars or derivatives thereof; L a and L b are the same or different and are each independently selected from any linking unit or bond; m is selected from an integer from 1 to 5; n is selected from an integer from 1 to 10; o is selected from integers from 1 to 10; A ligand drug conjugate, or a pharma- ceutically acceptable salt or solvate thereof, is provided.

[0005] In some embodiments, the ligand L is selected from an antibody, a functional antibody fragment, and a protein with a targeting effect. In some embodiments, the ligand L is selected from an antibody, including but not limited to a chimeric antibody, a humanized antibody, a fully human antibody, or a murine antibody.

[0006] In some embodiments, the linker unit M has the following formula: [ka] where the structure includes a cycloalkyl or heterocyclyl, the linker unit M may also be selected from derivatives thereof in which the cycloalkyl or heterocyclyl is in an open ring form; During the ceremony, The positions indicated by wavy lines are non-limiting links to ligands, linking scaffolds A, linking structures B, or branching units C; * is a chiral carbon and has an R or S configuration; p and p' are each independently selected from integers of 1 to 10; Ac is a residue of a natural or unnatural amino acid, a polyethylene glycol segment having 1 to 20 repeating units, a phosphate group, a carboxylate group, a sulfonate group, a sulfinate group, or the following structure: [ka] selected, but not limited to, from In the formula, the position indicated by the wavy line is * It is connected to the carbon atom at the position indicated by.

[0007] In some embodiments, the linker unit M has the following formula: [ka] or a stereoisomer or a ring-opened version thereof, or a derived structure thereof in which the ring of the succinimide group is in a ring-opened form, During the ceremony, The positions indicated by wavy lines are non-limiting links to ligands, linking scaffolds A, linking structures B, or branching units C; * is a chiral carbon and has an R or S configuration; p and p' are each independently selected from integers of 1 to 10; Ac is a residue of a natural or unnatural amino acid, a polyethylene glycol segment having 1 to 20 repeating units, a phosphate group, a carboxylate group, a sulfonate group, a sulfinate group, or the following structure: [ka] selected, but not limited to, from In the formula, the position indicated by the wavy line is * The carbon atom is connected to the carbon atom at the position indicated by the symbol:

[0008] In some embodiments, the linker unit M has the following formula: [ka] or a stereoisomer or derived structure thereof in which the ring of the succinimide group is open. In some embodiments, p is 1. In some embodiments, Ac is a glycine residue.

[0009] In some embodiments, the linker unit M has the following formula: [ka] or a derivative thereof in which the ring of the succinimide group is open. In some embodiments, the linker unit M is [ka] and; During the ceremony, * is a chiral carbon and has an R or S configuration; The positions indicated by wavy lines are non-limitingly linked to a ligand, a linking scaffold A, a linking structure B or a branching unit C.

[0010] In some embodiments, the linking scaffold A is comprised of one or more natural or unnatural amino acids or has the following structure: [ka] is selected from X is selected from N, CH, C3-C8 cycloalkyl, 3-8 membered heterocyclyl, aryl, substituted aryl, and heteroaryl; Y is -NH-, -O-, -S-, -CO-, -CO 2 -, -CONH-, -NHCO-, -SO-, -SO 2 -, -OSO 2-, and -OP=O(OH)O-; q is selected from an integer from 1 to 10; The positions indicated by wavy lines are linker unit M and linking unit L. a Or L b , linked, without limitation, to a linking structure B or a branching unit C; R 1 is selected from hydrogen, deuterium, halogen, C1-C6 alkyl, C1-C6 substituted alkyl, C3-C8 cycloalkylC1-C6 alkyl, C1-C6 alkoxyC1-C6 alkyl, carboxyl, 3-8 membered heterocyclyl, aryl, substituted aryl and heteroaryl; R 2 teeth, [ka] selected, but not limited to, from In the formula, the wavy line on the left is R 2 but * The wavy line on the right indicates one of three optional linking positions of the linking scaffold A. In some embodiments, linking scaffold A is [ka] In some embodiments, R 2 teeth, [ka] It is.

[0011] In some embodiments, linking scaffold A has the following structure: [ka] or stereoisomers thereof, During the ceremony, Z is selected from -NH-, -O-, and -S-; The positions indicated by wavy lines are linker unit M and linking unit L. a Or L b, linked to linking structure B or branching unit C, without any restriction. In some embodiments, linking scaffold A is [ka] In some embodiments, Z is -NH-. In some embodiments, the aryl is phenyl. In some embodiments, the substituted aryl is phenyl substituted with one or more substituents selected from deuterium, halogen, C1-C6 alkyl, C1-C6 alkoxy, and C1-C6 haloalkyl. In some embodiments, the heteroaryl is a 6-membered monoheteroaryl containing one or more heteroatoms selected from nitrogen, oxygen, and sulfur.

[0012] In some embodiments, B is selected from, but not limited to, any linking structure or bond. In some embodiments, C is present or absent, and if present, is selected from, but not limited to, one or more natural or unnatural amino acids. In some embodiments, drugs D, D 1 , and D 2 are the same or different and are each independently selected from, but not limited to, an anti-tumor drug, an autoimmune disease drug, an anti-infective drug (e.g., an anti-viral drug), a radioisotope, a chromogenic molecule, or a pharma- ceutically acceptable salt or solvate thereof. In some embodiments, drugs D, D 1 , and D 2 are the same or different and are independently selected, without limitation, from an anti-tumor drug, including, but not limited to, a DNA damaging drug, an RNA damaging drug, an enzyme inhibitor, or an anti-microtubule drug.

[0013] In some embodiments, the SU is selected from, but is not limited to, natural or unnatural monosaccharides, disaccharides, polysaccharides, and derivatives thereof. In some embodiments, the natural or unnatural monosaccharide, disaccharide, polysaccharide, or derivative thereof has the following structure: [ka] [ka] is selected, but not limited to, from In some embodiments, the SU is selected from methylglucamine, maltose, maltobionic acid, mannuronic acid, β-cyclodextrin, and mono(6-amino-6-deoxy)-β-cyclodextrin. In some embodiments, SU is covalently linked to L b is non-limitingly linked to

[0014] In some embodiments, the linking unit L a and L b are the same or different and are each independently selected from, but not limited to, one or more of a chemically labile linking unit, an enzyme-catalyzed cleavable linking unit, and a non-cleavable linking unit. In some embodiments, L a is selected from chemically labile linking units and enzyme-catalyzed cleavable linking units. In some embodiments, the linking unit L a has the following structure: [ka] or stereoisomers thereof, During the ceremony, R a , R b , and R c are the same or different and are each independently selected from hydrogen, deuterium, halogen, alkyl, substituted alkyl, deuterated alkyl, cycloalkyl, cycloalkylalkyl, alkoxyalkyl, heterocyclyl, aryl, substituted aryl, or heteroaryl; or R b , R c and R b and R c The carbon atoms linked to form a C3-C8 cycloalkyl or a 3-8 membered heterocyclyl; R d H, NO 2 , H.O.S.O.3 -and-OSO 3 Selected from; r is selected from an integer from 1 to 10; The position indicated by the wavy line on the left is linked to a linking scaffold A, a linking structure B or a branching unit C; The positions indicated by the wavy lines on the right are drugs D and D 1 Or D 2 is linked to.

[0015] In some embodiments, the linking unit L a -D, L a -D 1 and L a -D 2 are the same or different and have the following structure: (1) Auristatins [ka] (2) Maytansinoids [ka] (3) Benzodiazepines [ka] (4) Camptothecin analogues [ka] (5) Tubulysin [ka] (6) Adriamycin [ka] (7) Calicheamicin [ka] (8) Duocarmycin [ka] (9) Other antitumor drugs [ka] or stereoisomers thereof; During the ceremony, The positions indicated by wavy lines are linked to linking scaffold A, linking structure B or branching unit C.

[0016] In some embodiments, L a -D, L a -D 1 Or L a -D 2 are auristatins, e.g. [ka] , benzodiazepines, e.g. [ka] , camptothecin analogues, e.g. [ka] , combretastatins, e.g. [ka] is selected from.

[0017] In some embodiments, the linking unit L b has the following structure: [ka] or a stereoisomer thereof, but not limited to: During the ceremony, R 3 is selected from hydrogen, deuterium, halogen, C1-C6 alkyl, deuterated C1-C6 alkyl, halogenated C1-C6 alkyl, C3-C8 cycloalkylC1-C6 alkyl, C1-C6 alkoxyC1-C6 alkyl, heterocyclyl, aryl, substituted aryl, and heteroaryl; W is -NR 4-, -O-, -S-, -CO- and -CONR 5 - Selected from; R 4 or R 5 are independently selected from hydrogen, deuterium, halogen, C1-C6 alkyl, deuterated C1-C6 alkyl, halogenated C1-C6 alkyl, C3-C8 cycloalkylC1-C6 alkyl, C1-C6 alkoxyC1-C6 alkyl, heterocyclyl, aryl, substituted aryl, and heteroaryl; s is selected from an integer from 1 to 10; s' is selected from an integer from 1 to 10; The positions indicated by the left wavy line and the right wavy line are non-restrictively linked to SU or linking scaffold A.

[0018] The present application also relates to compounds of formula IV, V or VI: [ka] M is selected from any linker unit; A is selected from any linking scaffold; B is present or absent and, if present, is selected from any linking structure or bond; C is present or absent and, when present, is selected from any branching unit or bond; D, D 1 , D 2 are the same or different and are each independently selected from drugs; SU is selected from sugars or derivatives thereof; L a and L b are the same or different and are each independently selected from any linking unit or bond; m is selected from an integer from 1 to 5; o is selected from integers from 1 to 10; A linker drug compound or a pharma- ceutically acceptable salt or solvate thereof is provided.

[0019] In some embodiments, the linker unit M has the following formula: [ka] where the structure includes a cycloalkyl or heterocyclyl, the linker unit M may also be selected from derivatives thereof in which the cycloalkyl or heterocyclyl is in an open ring form; During the ceremony, The positions indicated by wavy lines are non-restrictively linked to linking scaffold A, linking structure B or branching unit C; M' is selected, but not limited to, from halogen, OTf and OTs; * is a chiral carbon and has an R or S configuration; p and p' are each independently selected from integers of 1 to 10; Ac is a residue of a natural or unnatural amino acid, a polyethylene glycol segment having 1 to 20 repeating units, a phosphate group, a carboxylate group, a sulfonate group, a sulfinate group, or the following structure: [ka] selected, but not limited to, from In the formula, the position indicated by the wavy line is * The carbon atom is connected to the carbon atom at the position indicated by the symbol:

[0020] In some embodiments, the linker unit M has the following formula: [ka] or a stereoisomer thereof or a derivative thereof in which the ring of the succinimide group is open, During the ceremony, The positions indicated by wavy lines are non-restrictively linked to linking scaffold A, linking structure B or branching unit C; * is a chiral carbon and has an R or S configuration; p and p' are each independently selected from integers of 1 to 10; Ac is a residue of a natural or unnatural amino acid, a polyethylene glycol segment having 1 to 20 repeating units, a phosphate group, a carboxylate group, a sulfonate group, a sulfinate group, or the following structure: [ka] selected, but not limited to, from In the formula, the position indicated by the wavy line is * The carbon atom is connected to the carbon atom at the position indicated by the symbol:

[0021] In some embodiments, the linker unit M has the following formula: [ka] or a derivative thereof in which the ring of the succinimide group is open. In some embodiments, the linker unit M is [ka] where: * is a chiral carbon and has an R or S configuration; The positions indicated by wavy lines are non-restrictively linked to linking scaffold A, linking structure B or branching unit C.

[0022] In some embodiments, the linking scaffold A is comprised of one or more natural or unnatural amino acids or has the following structure: [ka] is selected from the group consisting of X is selected from N, CH, C3-C8 cycloalkyl, 3-8 membered heterocyclyl, aryl, substituted aryl, and heteroaryl; Y is -NH-, -O-, -S-, -CO-, -CO 2 -, -CONH-, -NHCO-, -SO-, -SO 2 -, -OSO 2 -, and -OP=O(OH)O-; q is selected from an integer from 1 to 10; The positions indicated by wavy lines are linker unit M and linking unit L. a Or L b , linked, without limitation, to a linking structure B or a branching unit C; R 1 is selected from hydrogen, deuterium, halogen, C1-C6 alkyl, C1-C6 substituted alkyl, C3-C8 cycloalkylC1-C6 alkyl, C1-C6 alkoxyC1-C6 alkyl, carboxyl, 3-8 membered heterocyclyl, aryl, substituted aryl and heteroaryl; R 2 teeth, [ka] selected, but not limited to, from In the formula, the wavy line on the left is R 2 but * The wavy line on the right indicates one of three optional linking positions of the linking scaffold A.

[0023] In some embodiments, linking scaffold A has the following structure: [ka] or stereoisomers thereof, During the ceremony, Z is selected from -NH-, -O-, and -S-; The positions indicated by wavy lines are linker unit M and linking unit L. a Or L b , linked to linking structure B or branching unit C, without any restriction. In some embodiments, linking scaffold A is [ka] In some embodiments, Z is -NH-.

[0024] In some embodiments, B is selected from, but not limited to, any linking structure or bond. In some embodiments, C is present or absent and, if present, is selected from, but not limited to, one or more natural or unnatural amino acids. In some embodiments, drugs D, D 1 , and D 2 are the same or different and are each independently selected from, but not limited to, an anti-tumor drug, an autoimmune disease drug, an anti-infective drug (e.g., an anti-viral drug), a radioisotope, a chromogenic molecule, or a pharma- ceutically acceptable salt or solvate thereof. In some embodiments, drugs D, D 1 , and D 2 are the same or different and are independently selected, without limitation, from an anti-tumor drug, including, but not limited to, a DNA damaging drug, an RNA damaging drug, an enzyme inhibitor, or an anti-microtubule drug.

[0025] In some embodiments, the SU is selected from, but is not limited to, natural or unnatural monosaccharides, disaccharides, polysaccharides, and derivatives thereof. In some embodiments, the natural or unnatural monosaccharide, disaccharide, polysaccharide, or derivative thereof has the following structure: [ka] [ka] [ka] is selected, but not limited to, from In some embodiments, the SU is selected from methylglucamine, maltose, maltobionic acid, mannuronic acid, β-cyclodextrin, and mono(6-amino-6-deoxy)-β-cyclodextrin. In some embodiments, SU is covalently linked to L b is non-limitingly linked to

[0026] In some embodiments, the linking unit L a and L bare the same or different and are each independently selected from, but not limited to, one or more of a chemically labile linking unit, an enzyme-catalyzed cleavable linking unit, and a non-cleavable linking unit. In some embodiments, L a is selected from chemically labile linking units and enzyme-catalyzed cleavable linking units. In some embodiments, the linking unit L a has the following structure: [ka] or stereoisomers thereof, During the ceremony, R a , R b , and R c are the same or different and are each independently selected from hydrogen, deuterium, halogen, alkyl, substituted alkyl, deuterated alkyl, cycloalkyl, cycloalkylalkyl, alkoxyalkyl, heterocyclyl, aryl, substituted aryl, or heteroaryl; or R b , R c and R b and R c The carbon atoms linked to form a C3-C8 cycloalkyl or a 3-8 membered heterocyclyl; R d H, NO 2 , H.O.S.O. 3 and -OSO 3 Selected from; r is selected from an integer from 1 to 10; The position indicated by the wavy line on the left is linked to a linking scaffold A, a linking structure B or a branching unit C; The positions indicated by the wavy lines on the right are drugs D and D 1 Or D 2 is linked to.

[0027] In some embodiments, L a -D, L a -D 1 and L a -D 2 are the same or different and have the following structure: (1) Auristatins [ka] (2) Maytansinoids [ka] (3) Benzodiazepines [ka] (4) Camptothecin analogues [ka] (5) Tubulysin [ka] (6) Adriamycin [ka] (7) Calicheamicin [ka] (8) Duocarmycin [ka] (9) Other antitumor drugs [ka] or stereoisomers thereof; During the ceremony, The positions indicated by wavy lines are linked to linking scaffold A, linking structure B or branching unit C.

[0028] In some embodiments, L a -D, L a -D 1 Or L a -D 2 teeth, Auristatins, e.g. [ka] , benzodiazepines, e.g. [ka] , camptothecin analogues, e.g. [ka] , combretastatins, e.g. [ka] is selected from.

[0029] In some embodiments, the linking unit L b has the following structure: [ka] or stereoisomers thereof; During the ceremony, R 3 is selected from hydrogen, deuterium, halogen, C1-C6 alkyl, deuterated C1-C6 alkyl, halogenated C1-C6 alkyl, C3-C8 cycloalkylC1-C6 alkyl, C1-C6 alkoxyC1-C6 alkyl, heterocyclyl, aryl, substituted aryl, and heteroaryl; W is -NR 4 -, -O-, -S-, -CO- and -CONR 5 - Selected from; R 4 or R 5 are independently selected from hydrogen, deuterium, halogen, C1-C6 alkyl, deuterated C1-C6 alkyl, halogenated C1-C6 alkyl, C3-C8 cycloalkylC1-C6 alkyl, C1-C6 alkoxyC1-C6 alkyl, heterocyclyl, aryl, substituted aryl, and heteroaryl; s is selected from an integer from 1 to 10; s' is selected from an integer from 1 to 10; The positions indicated by the left wavy line and the right wavy line are non-restrictively linked to SU or linking scaffold A.

[0030] In some embodiments, L b -SU has the following structure: [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] is selected, but not limited to, from

[0031] In some embodiments, L b -SU has the following structure: [ka] [ka] is selected, but not limited to, from

[0032] In some embodiments, the linker drug compound has the following structure: [ka] [ka] [ka] [ka] [ka] [ka] is selected, but not limited to, from

[0033] In some embodiments, the linker drug compound has the following structure: [ka] [ka] [ka] [ka] is selected, but not limited to, from

[0034] In some embodiments, the linker drug compound has the following structure: [ka] [ka] [ka] is selected, but not limited to, from

[0035] In some embodiments, the linker drug compound has the following structure: [ka] [ka] [ka] is selected, but not limited to, from

[0036] The present application further relates to a method for preparing a ligand drug conjugate of formula I, formula II or formula III, or a pharma- ceutically acceptable salt or solvate thereof, comprising the steps of: [ka] reacting the modified ligand with a compound of formula IV, V, or VI to obtain a ligand drug conjugate of formula I, II, or III, or a pharma- ceutically acceptable salt or solvate thereof; During the ceremony, L is selected from a ligand; M is selected from any linker unit; A is selected from any linking scaffold; B is present or absent and, if present, is selected from any linking structure or bond; C is present or absent and, when present, is selected from any branching unit or bond; D, D 1 , D 2 are the same or different and are each independently selected from drugs; SU is selected from sugars or derivatives thereof; L a and L b are the same or different and are each independently selected from any linking unit or bond; m is selected from an integer from 1 to 5; n is selected from an integer from 1 to 10; o is selected from integers from 1 to 10; A method is provided.

[0037] With respect to the ligand drug conjugate of the present invention or a pharma- ceutically acceptable salt or solvate thereof, or a ligand drug conjugate comprising a linker drug compound of the present invention or a pharma- ceutically acceptable salt or solvate of said ligand drug conjugate, in some embodiments the ligand drug conjugate or a pharma- ceutically acceptable salt or solvate thereof has the following structure: [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] or stereoisomers thereof, During the ceremony, L is selected from a ligand, preferably an antibody; n is selected from integers of 1 to 10, preferably 2 to 8.

[0038] In some embodiments, the pharma- ceutically acceptable salt is Sodium salts, potassium salts, calcium salts, or magnesium salts formed by the acidic functional groups in the above structural formula and alkali; and Acetate, trifluoroacetate, citrate, oxalate, tartrate, malate, nitrate, chloride, bromide, iodide, sulfate, bisulfate, phosphate, lactate, oleate, ascorbate, salicylate, formate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, or p-toluenesulfonate formed by a basic functional group in the structure and an acid. is selected from.

[0039] In some embodiments, the ligand is selected from a monoclonal antibody, an anti-EGFRvIII antibody, an anti-DLL-3 antibody, an anti-PSMA antibody, an anti-CD70 antibody, an anti-MUC16 antibody, an anti-ENPP3 antibody, an anti-TDGF1 antibody, an anti-ETBR antibody, an anti-MSLN antibody, an anti-TIM-1 antibody, an anti-LRRC15 antibody, an anti-LIV-1 antibody, an anti-CanAg / AFP antibody, an anti-claudin 18.2 antibody, an anti-mesothelin antibody, an anti-HER2 (ErbB2) antibody, an anti-EGFR antibody, an anti-C-Met antibody, an anti-SLITRK6 antibody, an anti-KIT / CD117 antibody, an anti-STEAP1 antibody, an anti-SLAMF7 / CS1 antibody, an anti-NaPi2B / SLC34A2 antibody, an anti-GPNMB antibody, an anti-HER3 (ErbB3) antibody, an anti-MUC1 / CD227 antibody, an anti-AXL antibody, an anti-CD166 antibody, an anti-B7-H3 (CD276) antibody, an anti-PTK7 / C CK4 antibody, anti-PRLR antibody, anti-EFNA4 antibody, anti-5T4 antibody, anti-NOTCH3 antibody, anti-Nectin 4 antibody, anti-Trop-2 antibody, anti-CD142 antibody, anti-CA6 antibody, anti-GPR20 antibody, anti-CD1 74 antibody, anti-CD71 antibody, anti-EphA2 antibody, anti-LYPD3 antibody, anti-FGFR2 antibody, anti-FGFR3 antibody, anti-FRα antibody, anti-CEACAMs antibody, anti-GCC antibody, anti-integrin Av antibody, anti-C The antibody may be selected from, but is not limited to, an AIX antibody, an anti-P-cadherin antibody, an anti-GD3 antibody, an anti-cadherin 6 antibody, an anti-LAMP1 antibody, an anti-FLT3 antibody, an anti-BCMA antibody, an anti-CD79b antibody, an anti-CD19 antibody, an anti-CD33 antibody, an anti-CD56 body antibody, an anti-CD74 antibody, an anti-CD22 antibody, an anti-CD30 antibody, an anti-CD37 antibody, an anti-CD138 antibody, an anti-CD352 antibody, an anti-CD25 antibody, and an anti-CD123 antibody. In some embodiments, the ligand is an anti-Trop-2 antibody, for example the light and heavy chain sequences of which are shown in SEQ ID NO: 1 and SEQ ID NO: 2, respectively. In some embodiments, the ligand is an anti-CD33 antibody, for example the light and heavy chain sequences of the anti-CD33 antibody are set forth in SEQ ID NO: 3 and SEQ ID NO: 4, respectively.

[0040] The present application further provides a pharmaceutical composition comprising a therapeutically effective amount of a ligand drug conjugate of the present invention or a pharma- ceutically acceptable salt of said ligand drug conjugate, or a linker drug compound of the present invention or a pharma- ceutically acceptable salt or solvate of said linker drug compound, and a pharma- ceutically acceptable carrier, diluent or excipient. The present application further provides the use of a ligand drug conjugate of the present invention or a pharma- ceutically acceptable salt of said ligand drug conjugate, or a linker drug compound of the present invention or a pharma- ceutically acceptable salt or solvate of said linker drug compound, in the preparation of a drug for treating or preventing a tumor or an autoimmune disease. In some embodiments, the tumor is selected from a solid tumor or a non-solid tumor, such as breast cancer, ovarian cancer, cervical cancer, uterine cancer, prostate cancer, kidney cancer, urethral cancer, bladder cancer, liver cancer, stomach cancer, endometrial cancer, salivary gland cancer, esophageal cancer, lung cancer, colon cancer, rectal cancer, colorectal cancer, bone cancer, skin cancer (e.g., epidermal cell carcinoma of the skin (also known as squamous cell carcinoma of the skin)), thyroid cancer, pancreatic cancer, melanoma, glioma, neuroblastoma, glioblastoma multiforme, sarcoma, lymphoma, and leukemia (e.g., acute and chronic leukemias, such as acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), chronic lymphocytic leukemia (CLL), hairy cell leukemia, and prolymphocytic leukemia).

[0041] definition Unless otherwise defined, all technical and scientific terms used herein are consistent with the common understanding of one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used to practice or test the present invention, the present invention describes preferred methods and materials. In describing and claiming the present invention, the following terms are used in accordance with the following definitions. When a trademark is used herein, applicants intend to include formulations of that trademarked product, generic versions of that trademarked product, and active pharmaceutical ingredients. Unless otherwise stated, terms used in the specification and claims have the following meanings:

[0042] The term "ligand" refers to a targeting agent that specifically binds to a target moiety. A ligand can specifically bind to a cellular component or other target molecule of interest. The target moiety or target is usually on the cell surface. In some embodiments, the ligand serves to deliver the drug unit to a specific target cell population with which the ligand unit interacts. Ligands include, but are not limited to, proteins, polypeptides and peptides, as well as non-proteins such as sugars. Suitable ligand units include, for example, antibodies, such as full-length (intact) antibodies and antigen-binding fragments thereof. In embodiments where the ligand unit is a non-antibody targeting agent, it can be a peptide or polypeptide, or a non-protein molecule. Examples of such targeting agents include interferons, lymphokines, hormones, growth factors and colony stimulating factors, vitamins, nutrient transport molecules, or any other cell-binding molecule or substance. In some embodiments, the linker is covalently bonded to a sulfur atom of the ligand. In some embodiments, the sulfur atom is of a cysteine ​​residue that forms an intrachain disulfide bond of an antibody. In another embodiment, the sulfur atom is the sulfur atom of a cysteine ​​residue that has been introduced into the Ligand unit, which sulfur atom forms an intrachain disulfide bond of the antibody, hi another embodiment, the sulfur atom is the sulfur atom of a cysteine ​​residue that has been introduced into the Ligand unit, for example, by site-directed mutagenesis or chemical reaction.

[0043] The term "drug" refers to a cytotoxic drug, i.e., a molecule that has a strong ability to destroy the normal proliferation of tumor cells or cancer cells. In principle, cytotoxic drugs can kill tumor cells at a sufficiently high concentration, but due to lack of specificity, while killing tumor cells or cancer cells, they may also cause apoptosis of normal cells, and thus are prone to cause severe side effects. The term "ligand drug conjugate" refers to a molecule formed by linking a ligand and a drug by a stable linking unit. In the present invention, the "ligand drug conjugate" is preferably an antibody drug conjugate (ADC), which refers to a monoclonal antibody or a functional antibody fragment or a target protein and a cytotoxic drug linked by a stable linking unit. The term "antibody" or "functional antibody fragment" includes within its scope any part of the antibody structure, which unit is capable of binding, reactively associating with or forming a complex with a receptor, antigen, or other receptor unit present in the target cell population. An antibody may be any protein or proteinaceous molecule that binds, complexes, or reacts with a portion of the cell population to be treated or biomodified. The antibodies of the present invention include, but are not limited to, murine antibodies, chimeric antibodies, humanized antibodies and fully human antibodies, preferably humanized antibodies and fully human antibodies.

[0044] The term "linker unit" refers to a chemical fragment or bond that is linked at one end to a ligand and at the other end to a drug, either directly or to another linker, which is then linked to a drug. In the general formulas I-VI of the present application, the linker unit is represented by M. The term "linking scaffold" refers to a chemical structure fragment or bond having a branched structure that can be linked to a linker unit, a ligand, a drug, or a hydrophilic unit, etc., and the linking scaffold is represented by A. The linking mode of the present invention is preferably a covalent linkage. The term "linking unit" refers to a chemical structure fragment or bond that can link different components in an antibody-drug conjugate or linker-drug compound, such as a branching unit, a linking structure, a drug or a hydrophilic unit. According to the drug release mechanism in cells, the linking units can be divided into two categories: non-cleavable linkers and cleavable linkers, and the linking units are L, L a , and L b It is expressed as:

[0045] For ligand-drug conjugates containing non-cleavable linkers, the drug release mechanism is as follows: after the conjugate binds to the antigen and is taken up by the cell by endocytosis, the antibody is enzymatically hydrolyzed in the lysosome, releasing the active molecule consisting of the small molecule drug, the linker, and the amino acid residues of the antibody. As a result, the cytotoxicity of the drug molecule is not weakened even if the structure of the drug molecule is changed, but the active molecule is charged (amino acid residues) and therefore cannot penetrate into neighboring cells. Therefore, such an active drug cannot kill adjacent tumor cells that do not express the target antigen (antigen-negative cells) (bystander effect) (Ducry et al., 2010, Bioconjugate Chem. 21: 5-13). As the name suggests, cleavable linkers can be cleaved in target cells to release the active drug (the small molecule drug itself). Cleavable linkers can be divided into two main categories: chemically labile linkers and enzyme-labile linkers. Chemically labile linkers are selectively cleavable due to differences in plasma and cytoplasmic properties, including pH and glutathione concentration. pH-sensitive linkers are often called acid-cleavable linkers. Such linkers are relatively stable in the neutral environment of blood (pH 7.3-7.5), but are hydrolyzed in the weakly acidic environment of endosomes (pH 5.0-6.5) and lysosomes (pH 4.5-5.0). Most of the first generation antibody-drug conjugates used this type of linker, such as hydrazones, carbonates, acetals, and ketals. Because of the limited plasma stability of acid-cleavable linkers, the half-life of antibody-drug conjugates based on this type of linker is usually short (2-3 days). This short half-life limits the application of pH-sensitive linkers to new generation antibody-drug conjugates to some extent.

[0046] Glutathione-sensitive linkers are also called disulfide linkers. Drug release is triggered by the difference between high intracellular glutathione concentrations (millimolar range) and relatively low blood glutathione concentrations (micromolar range). This is especially true in tumor cells, where low oxygen content leads to increased activity of reductase enzymes and therefore higher glutathione concentrations. Disulfide bonds are thermodynamically stable, so stability in plasma is good. Enzyme-labile linkers, such as peptide linkers, allow for better control of drug release. Peptide linkers can be effectively cleaved by lysosomal proteases, such as cathepsin B or plasmin (some tumor tissues have increased levels of such enzymes). Such peptide linkers are considered to be very stable in plasma circulation, since proteases are usually inactive due to inappropriate extracellular pH and serum protease inhibitors. Enzyme-labile linkers are widely used as cleavable linkers in antibody-drug conjugates, due to their high plasma stability and good selectivity and efficacy of intracellular cleavage. Representative enzyme-labile linkers include Val-Cit (VC), Phe-Lys, etc. The suicide linker may be embedded between the cleavable linker and the effective drug, or the suicide linker itself may be part of the cleavable linker. The mechanism of action of the suicide linker is as follows: when the cleavable linker is cleaved under appropriate conditions, the suicide linker can spontaneously rearrange its structure and release the effective drug linked to it. Common suicide linkers include p-aminobenzyl alcohol (PAB) and β-glucuronide.

[0047] The term "branching unit" or "linking structure" refers to a chemical structure fragment having a branching structure or bond that connects different components within a ligand drug conjugate. The linking mode of the present invention is preferably a covalent linkage. The term "sugar" refers to a polyhydroxy aldehyde or ketone compound that can be decomposed into one of polyhydroxy aldehyde and ketone compounds after hydrolysis. Sugars can be divided into monosaccharides, disaccharides and polysaccharides. The three-letter and one-letter codes for amino acids used in the present invention are as described in J. Boil. Chem. 1968, 243, 3558. The term "natural amino acid" refers to an amino acid that can be synthesized by living organisms. Natural amino acids are generally in the L-form, although there are some exceptions, such as glycine, which includes natural and biosynthetic glycine. The term "unnatural amino acid" refers to amino acids that can only be synthesized artificially.

[0048] The term "alkyl" refers to saturated aliphatic hydrocarbon groups, i.e., straight or branched chain groups containing from 1 to 20 carbon atoms, preferably alkyl containing from 1 to 12 carbon atoms, more preferably alkyl containing from 1 to 10 carbon atoms, and most preferably alkyl containing from 1 to 6 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, Examples of dimethylhexyl include 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched isomers thereof.More preferably, the alkyl group is a lower alkyl having 1-6 (e.g., 1-4) carbon atoms, non-limiting examples of which include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and the like. The alkyl may be substituted or unsubstituted. If substituted, the substituent may be substituted at any available attachment point. The substituents are preferably one or more groups independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, thiol, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, and oxo.

[0049] The term "substituted alkyl" means that a hydrogen in an alkyl is replaced by a substituent. Unless otherwise specified in the context, the substituents of an alkyl are -halogen, -OR', -NR'R'', -SR', -SiR'R''R''', -OC(O)R', -C(O)R', -CO 2 R', -CONR'R'', -OC(O)NR'R'', -NR''C(O)R', -NR'-C(O)NR''R''', -NR''C(O) 2 R', -NH-C(NH 2 )=NH, -NR'C(NH 2 )=NH, -NH-C(NH 2 )=NR', -S(O)R', -S(O) 2 R', -S(O) 2 NR'R'', -NR'S(O) 2 R'', -CN and -NO2 and the number of substituents ranges from 1 to (2m'+1), where m' is the total number of carbon atoms in the group, such as 1, 2, 3, 4, 5, or 6. R', R'', and R''' are each independently hydrogen, C 1-8 Alkyl, aryl, aryl substituted with 1-3 halogens, C substituted with 1-3 halogens 1-8 Alkyl, C 1-8 Alkoxy or C 1-8 Thioalkoxy or unsubstituted aryl-C 1-4 When R' and R" are attached to the same nitrogen atom, they can be combined with the nitrogen atom to form a 3-, 4-, 5-, 6-, or 7-membered ring. For example, -NR'R" includes 1-pyrrolidinyl and 4-morpholinyl.

[0050] The term "cycloalkyl" refers to a hydrocarbon group of saturated or partially unsaturated monocyclic or polycyclic configuration, the ring of the cycloalkyl containing from 3 to 20 carbon atoms, preferably from 3 to 12 carbon atoms, more preferably from 3 to 10 carbon atoms, and most preferably from 3 to 8 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, and cyclooctyl. Non-limiting examples of polycyclic cycloalkyls include spiro, fused, and bridged cycloalkyl groups. The term "cycloalkylalkyl" means that an alkyl is substituted with one or more cycloalkyl groups, preferably one cycloalkyl group, where alkyl is as defined above and cycloalkyl is as defined above. The cycloalkyl ring preferably contains 3 to 12 carbon atoms, more preferably 3 to 10 carbon atoms, and most preferably 3 to 8 carbon atoms. The alkyl is preferably a lower alkyl having 1 to 6 carbon atoms, more preferably 1 to 4 carbon atoms.

[0051] The term "alkoxy" refers to -O-(alkyl) and -O-(cycloalkyl) groups, where alkyl and cycloalkyl are as defined above. Non-limiting examples of alkoxy include methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, and cyclohexyloxy. An alkoxy may be optionally substituted or unsubstituted. If substituted, the substituents are preferably one or more groups independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, thiol, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, and heterocycloalkylthio. The term "heterocyclyl" refers to a heterocyclic ring containing 3 to 20 ring atoms, one or more (e.g., 1, 2, 3, or 4) of which are nitrogen, oxygen, or S(O). m (wherein m is 0, 1 or 2), with the remaining ring atoms being carbon. Preferably, the heterocyclyl contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms. More preferably, the heterocyclyl contains 3 to 10 ring atoms. Non-limiting examples of monocyclic heterocyclyls include pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, and homopiperazinyl. Non-limiting examples of polycyclic heterocyclyls include spiro, fused, and bridged heterocyclyls. The term "aryl" refers to a 6-14 membered all carbon monocyclic or fused polycyclic (i.e. rings sharing adjacent pairs of carbon atoms) group having a conjugated π electron system, preferably a 6-10 membered group, such as phenyl. Aryl can be substituted or unsubstituted. When aryl is substituted, the substituents are preferably one or more of the following groups selected from, but not limited to, alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, deuterium atom, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, and heterocycloalkylthio.

[0052] The term "heteroaryl" includes 5- to 8-membered monocyclic heteroaryls and 8- to 12-membered fused heteroaryls. The term "5-8 membered monocyclic heteroaryl" refers to an aromatic monocyclic group containing 5-8 ring atoms (at least one of which is a heteroatom such as a nitrogen atom, an oxygen atom, or a sulfur atom). Optionally, a ring atom (e.g., a carbon atom, a nitrogen atom, or a sulfur atom) in the ring structure may be oxo-substituted. "5-8 membered monocyclic heteroaryl" includes, for example, "5-7 membered monocyclic heteroaryl", "5-6 membered monocyclic heteroaryl", "5-6 membered monocyclic nitrogen-containing heteroaryl", and "6 membered monocyclic nitrogen-containing heteroaryl", and the heteroatom in "nitrogen-containing heteroaryl" includes at least one nitrogen atom, for example, only one or two nitrogen atoms, or includes one nitrogen atom and one or two other heteroatoms (e.g., oxygen atom and / or sulfur atom), or includes two nitrogen atoms and one or two other heteroatoms (e.g., oxygen atom and / or sulfur atom). Specific examples of the "5- to 8-membered monocyclic heteroaryl" include, but are not limited to, furyl, thienyl, pyrrolyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, imidazolyl, pyrazolyl, 1,2,3-triazole base, 1,2,4-triazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, pyridyl, 2-pyridonyl, 4-pyridonyl, pyrimidinyl, pyridazinyl, pyrazinyl, 1,2,3-triazinyl, 1,3,5-triazine base, 1,2,4,5-tetrazinyl, azepantrienyl, 1,3-diazacycloheptatrienyl, and azepantetraenyl.

[0053] The term "8-12 membered fused heteroaryl" refers to an unsaturated aromatic ring structure containing 8 to 12 ring atoms (at least one of which is a heteroatom such as a nitrogen atom, an oxygen atom, or a sulfur atom) formed by two or more ring structures sharing two adjacent atoms. Optionally, a ring atom (e.g., a carbon atom, a nitrogen atom, or a sulfur atom) in the ring structure may be oxo-substituted. The "8- to 12-membered fused heteroaryl" includes "8- to 10-membered fused heteroaryl" and "8- to 9-membered fused heteroaryl"; specific examples include, but are not limited to, pyrrolopyrrole, pyrrolofuran, pyrazopyrrole, pyrazothiophene, furothiophene, pyrazoxazole, benzofuryl, benzisofuryl, benzothienyl, indolyl, isoindolyl, benzoxazolyl, benzimidazolyl, indazolyl, benzotriazolyl, quinolinyl, 2-quinolinonyl, 4-quinolinonyl, 1-isoquinolinonyl, isoquinolinyl, acridinyl, phenanthridinyl, benzopyridazinyl, phthalazinyl, quinazolinyl, quinoxalinyl, purinyl, and naphthyridinyl.

[0054] The term "haloalkyl" refers to an alkyl substituted with one or more halogens, where alkyl is defined above. The term "deuterated alkyl" refers to an alkyl substituted with one or more deuterium atoms, where alkyl is defined above. The term "hydroxy" refers to an --OH group. The term "halogen" refers to fluorine, chlorine, bromine or iodine. The term "amino" means -NH 2 Refers to the base. The term "nitro" means -NO 2 Refers to the base. The term "derivative" refers to a substance that has a similar chemical structure to that of a compound, but also contains at least one chemical group not present in the compound and / or lacks at least one chemical group present in the compound. The compound to which the derivative is compared is called the "parent" compound. In general, a "derivative" can be produced from the parent compound in one or more chemical reaction steps.

[0055] The term "pharmaceutical acceptable salt" refers to a pharmaceutical acceptable organic or inorganic salt of a compound (e.g., a drug, a linker-drug compound, or a ligand-drug conjugate). The compound or conjugate may contain at least one amino or carboxyl group, and thus may form an addition salt with the corresponding acid or alkali. Exemplary salts include, but are not limited to, sulfate, trifluoroacetate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, perphosphate, isonicotinate, lactate, salicylate, percitrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, salicylate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, potassium salt, and sodium salt. The term "solvate" refers to a compound formed by a linker drug compound or a ligand drug conjugate of the invention with one or more solvent molecules, including but not limited to water, ethanol, acetonitrile, isopropanol, DMSO, and ethyl acetate.

[0056] The term "pharmaceutical composition" refers to a mixture containing one or more compounds of the present invention or their physiologically / pharmaceutical acceptable salts or prodrugs and other chemical components, as well as other components such as physiologically / pharmaceutical acceptable carriers and / or excipients. The purpose of a pharmaceutical composition is to facilitate drug administration to an organism and facilitate absorption of the active ingredient, thus exerting biological activity. The term "carrier" refers to a system that can change the way drugs enter the human body and their distribution in the body, control the release rate of drugs, and deliver drugs to a target. Drug carrier release and targeting systems can reduce the degradation and loss of drugs, reduce side effects, and improve bioavailability. The term "excipient" refers to additives or vehicles other than the main drug in a pharmaceutical formulation. For example, adhesives, fillers, disintegrants, and lubricants in tablets; matrix back-ups in semi-solid formulations such as ointments and creams; preservatives, antioxidants, flavors, perfumes, cosolvents, emulsifiers, permeation enhancers, osmotic pressure regulators, and colorants in liquid formulations can all be called excipients. The term "diluent" or "filler" is primarily used to increase the mass and / or volume of a formulation. The addition of a diluent not only ensures a constant volume, but also reduces the dosage deviation of the main ingredient and improves the compression molding of the drug.

[0057] In one embodiment of the present invention, the linker drug compound is linked to the ligand via a thiol group generated by an open interchain disulfide bond or a cysteine ​​thiol group introduced by genetic engineering. In general, the number of drug molecules that can be linked to the ligand in a conjugation reaction will be less than or equal to the theoretical maximum. The drug loading of the ligand drug conjugate is 1) controlling the molar ratio of linking agent to ligand; 2) Controlling reaction time and temperature 3) Selecting different reaction reagents The amount of oxygen can be controlled by a number of methods, including but not limited to: The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments. It should be understood that these embodiments are only used to explain the present invention, and are not used to limit the scope of the present invention. In the following embodiments, experimental methods that do not specify specific conditions are usually carried out under conventional conditions or conditions recommended by the manufacturer. Unless otherwise specified, percentages, proportions, ratios, or parts are all calculated by mass. [Brief description of the drawings]

[0058] [Figure 1A] FIG. 1A is a SEC-HPLC spectrum of ADC-29, showing aggregation of ADC-29. [Figure 1B] FIG. 1B is a SEC-HPLC spectrum of ADC-56, showing aggregation of ADC-56. [Figure 1C] FIG. 1C is a SEC-HPLC spectrum of ADC-111, showing aggregation of ADC-111. [Figure 2A] FIG. 2A is an RP-HPLC spectrum of ADC-29 showing the drug / antibody ratio (DAR) of ADC-29. [Figure 2B] FIG. 2B is an RP-HPLC spectrum of ADC-56, showing the DAR of ADC-56. [Figure 2C] FIG. 2C is an RP-HPLC spectrum of ADC-111, showing the DAR of ADC-111. [Figure 3A] FIG. 3A shows the in vitro efficacy of ADC-1, ADC-28, ADC-29, ADC-42, ADC-56, ADC-109, and ADC-111 in the A431 cell line. [Figure 3B] FIG. 3B shows the in vitro efficacy of ADC-1, ADC-28, ADC-29, ADC-42, ADC-56, ADC-109, and ADC-111 in the HL-60 cell line. [Figure 3C] FIG. 3C shows the in vitro efficacy of ADC-1, ADC-28, ADC-29, ADC-42, ADC-56, ADC-109, and ADC-111 in the TF-1 cell line. [Figure 3D] FIG. 3D shows the in vitro efficacy of ADC-1, ADC-28, ADC-29, ADC-42, ADC-56, ADC-109, and ADC-111 in the HEL92.1.7 cell line. [Figure 3E]FIG. 3E shows the in vitro efficacy of ADC-1, ADC-28, ADC-29, ADC-42, ADC-56, ADC-109, and ADC-111 in the MV4-11 cell line. [Figure 3F] FIG. 3F shows the in vitro efficacy of ADC-1, ADC-28, ADC-29, ADC-42, ADC-56, ADC-109, and ADC-111 in MOLM-13 cell lines. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0059] Detailed Description of the Preferred Embodiments Example 1: Synthesis of Compound 6 [ka] Add 400 mL of benzyl alcohol to a 1 L single-neck flask, then add SOCl 2 (13.38mL, 184.8mmol) was slowly added dropwise under cooling in a water bath, and the reaction was left at the water bath temperature for 1 hour. Compound (S)-2-(CBZ-amino)-3-aminopropionic acid (40g, 168mmol) was added in three portions, and the resulting reaction solution was reacted at room temperature overnight. After the reaction was completed, the unreacted benzyl alcohol was removed by vacuum evaporation using an oil pump, and the resulting brown residue was purified by silica gel column chromatography (DCM:MeOH=150:1) to obtain 44g of compound 1. LC-MS m / z (M+H) + :329.1. Compound 1 (40 g, 121.6 mmol) was dissolved in 600 mL of acetonitrile, and then DIPEA (18.88 mL, 146.32 mmol) was added under cooling in an ice-water bath. tert-Butyl bromoacetate (23.9 g, 120 mmol) was added dropwise under low temperature conditions, and the resulting reaction solution was stirred for 30 minutes, and then transferred to room temperature for reaction. After the reaction was found to be complete by monitoring TLC (DCM:MeOH=10:1), the reaction solution was concentrated under reduced pressure in a water bath, and the resulting residue was purified by silica gel column chromatography (dichloromethane:methanol=100:1) to obtain 45.5 g of compound 2. LC-MS m / z (M+H) + :443.3.

[0060] In a 500 mL one-necked reaction flask, compound 2 (45 g, 25.2 mmol), 100 mL of 1,4-dioxane and 100 mL of water were mixed, and then DIPEA (25.2 mL, 152.5 mmol) was added. Boc anhydride (110.6 g, 508.7 mmol) was added dropwise at room temperature to obtain a yellow reaction solution, and the reaction was allowed to stand at room temperature for about 3-4 hours. After the reaction was found to be complete by monitoring TLC (DCM:MeOH=30:1), the reaction solution was concentrated under reduced pressure in a water bath to remove dioxane and extracted with DCM (100 mL x 3). The resulting organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a yellow oily crude product. The crude product was purified by silica gel column chromatography (DCM:MeOH=300:1) to obtain 50.3 g of compound 3. LC-MS m / z (M+H) + :543.2. 50g of compound 3 was dissolved in 250mL of methanol, then 10g of 5% Pd / C was added, the atmosphere of the system was replaced with hydrogen 2-3 times, and the reaction was left at room temperature. After the reaction was found to be complete by monitoring TLC (DCM:MeOH=5:1), the reaction solution was filtered through two pieces of filter paper. The filter cake was rinsed with methanol three times, and the obtained filtrate was concentrated under reduced pressure at 40°C to obtain compound 4, which was sent directly to the next reaction without purification. Crude compound 4 was dissolved in 250 mL of glacial acetic acid, maleic anhydride (18.2 g, 184.4 mmol) was added, and the reaction solution was reacted at room temperature with stirring. After the reaction was found to be complete by monitoring TLC (DCM:MeOH=3:1), the reaction solution was directly concentrated under reduced pressure, and the residue was purified by preparative high performance liquid chromatography; Column: YMC-C18, 100 mm * Tested at 450 mm, 10 μm, acetonitrile / water (0.2% TFA), flow rate 200 mL / min, λ = 215 nm; Solvent A: 0.2% TFA in water; Solvent B: acetonitrile; Gradient: 0~10 min 90%A, 10~25 min 90%A~45%A, 25~55 min 45%A~40%A; The fraction with a retention time of 43 min was collected and lyophilized to give 41 g of compound 5 as a white solid. LC-MS m / z (M+H) + :417.9.

[0061] In a 1 L single-neck flask, compound 5 (40 g, 96 mmol, 1.0 equiv), triethylamine (26.7 mL, 2.0 equiv), and toluene (400 mL) were mixed and then heated to 120 °C and refluxed for 2 h. After TLC monitoring showed the reaction was complete, the reaction was cooled to 50 °C and rotary evaporated under reduced pressure to remove the solvent. The reaction product was then dissolved in ethyl acetate (150 mL) and water (40 mL), and then 1 M HCl was added with stirring in an ice bath to adjust the pH to 2-3. The resulting solution was allowed to stand and the liquids were separated; the aqueous layer was extracted once more with ethyl acetate; the organic layers were combined and washed with anhydrous NaCl. 2 SO 4 After filtration, the filtrate was concentrated to give a crude product as a pale yellow oil. The crude product was purified by column chromatography (DCM:MeOH=40:1) to give 26.6 g of compound 6; LC-MS m / z (M+H) + :399.3.

[0062] Example 2: Synthesis of Compound 9 [ka] Into a 1L single-neck flask, 1-tert-butyl N-Fmoc-L-glutamate (21 g, 48.9 mmol), DCC (20.6 g, 97.8 mmol), Val-Cit-PABOH (18.5 g, 48.9 mmol) and dichloromethane (400 mL) were added, and then the resulting solution was cooled to 0° C. under ice-water bath conditions; 4-DMAP (610 mg, 4.89 mmol) was weighed and added to the reaction flask in one portion, and the resulting solution was allowed to slowly warm to room temperature and react for 3 hours. After the reaction was found to be complete by TLC monitoring, the insoluble solid impurities were filtered, the filter cake was rinsed with dichloromethane three times, and the solvent was removed by rotary evaporation under reduced pressure, and the residue was concentrated to obtain a pale yellow oily crude product. The crude product was purified by column chromatography (DCM:MeOH=100:1 to 50:1) to give 33.2 g of compound 7; LC-MS m / z (M+H) + :787.5. The above compound 7 was dissolved in 300 mL of dichloromethane, and then the reaction solution was cooled to 0° C. under ice-water bath conditions, TFA (150 mL) was slowly added dropwise, and the resulting solution was allowed to slowly come to room temperature and react for 5 hours. After the reaction was found to be complete by monitoring TLC (DCM:MeOH=10:1), the reaction solution was concentrated under reduced pressure at 45° C. to obtain a crude product of yellow oily compound 8, which was directly sent to the next reaction without purification.

[0063] In a 1 L single neck flask, the above crude compound 8 (48.9 mmol based on the starting material charge), PyBOP (38.56 g, 73.4 mmol), propargylamine (3.3 g, 58.7 mmol) and dichloromethane (400 mL) were mixed and then cooled to 0 °C under ice-water bath conditions; DIPEA (2.6 mL, 14.7 mmol) was added dropwise to the reaction flask and the reaction was allowed to stand at room temperature overnight. After the reaction was found to be complete by TLC (20:1) monitoring, the reaction solution was diluted with saturated aqueous NaCl (100 mL) and extracted twice with dichloromethane (100 mL); the organic liquid was combined and washed with anhydrous Na2 SO 4 The mixture was dried at 40° C., filtered by suction, and rotary evaporated under reduced pressure to remove the solvent. The residue was concentrated to give a light yellow oily crude product, which was purified by column chromatography (DCM:MeOH=50:1) to give 25.4 g of compound 9; LC-MS m / z (M+H) + :768.4.

[0064] Example 3: Synthesis of Compound 10 [ka] At room temperature, maltobionic acid (7.0 g, 19.5 mmol) was dissolved in 70 mL of anhydrous methanol and the atmosphere of the system was purged with N 2 The reaction mixture was replaced with 1-amino-11-azido-3,6,9-trioxaundecane three times. 4 Å molecular sieves were added, and the reaction solution was heated to reflux overnight and then cooled to room temperature. 1-Amino-11-azido-3,6,9-trioxaundecane was added and stirred at room temperature for 24 hours. The reaction mixture was concentrated under reduced pressure and the crude product obtained was dissolved in 5 mL of methanol, followed by the addition of 30 mL of chloroform and 100 mL of tetrahydrofuran. The resulting solution was stirred and allowed to stand to give a white solid, which was then filtered and the filter cake was rinsed twice with tetrahydrofuran. The resulting white solid was dissolved in 30 mL of water and lyophilized to give 5.6 g of white solid compound 10; LC-MS m / z (M+H) + :559.3.

[0065] Example 4: Synthesis of Compound 12 [ka] Compound 9 (5.0 g, 6.5 mmol) and dichloromethane (100 mL) were added to a 250 mL single-neck flask at room temperature, followed by NPC (10.1 g, 32.6 mmol) and DIPEA (5.7 mL, 32.6 mmol) in sequence, and the reaction was allowed to stand at room temperature overnight. After TLC monitoring showed the reaction was complete, the reaction solution was diluted with saturated aqueous NaCl (50 mL) and extracted twice with dichloromethane (100 mL). The organic liquids were combined and washed with anhydrous Na 2 SO 4 The mixture was dried at 40° C. and filtered by suction. The solvent was removed by rotary evaporation under reduced pressure, and the residue was concentrated to give a light yellow oily crude product. The crude product was purified by column chromatography (DCM:MeOH=100:1 to 50:1) to give 5.3 g of compound 11; LC-MS m / z (M+H) + :933.5. Compound 11 (1.0 g, 1.1 mmol) and DMF (15 mL) were added to a 50 mL single-neck flask at room temperature, followed by the addition of MMAE (723 mg, 1 mmol), HOBt (304 mg, 2.2 mmol) and DIPEA (390 μL, 2.2 mmol) in sequence, and the reaction was allowed to stand at room temperature overnight. After the reaction was completed, the reaction solution was directly concentrated under reduced pressure using an oil pump. The residue was purified by preparative high performance liquid chromatography to give 1.09 g of compound 12 as a white solid; TOF-MS m / z (M+Na) + :1534.0.

[0066] Example 5: Synthesis of Compound 15 [ka] Compound 7 (10.0 g, 12.7 mmol) and dichloromethane (130 mL) were added to a 250 mL one-neck flask at room temperature, followed by NPC (19.7 g, 63.5 mmol) and DIPEA (11.1 mL, 63.5 mmol) in sequence, and the reaction was allowed to proceed at room temperature overnight. After TLC monitoring showed that the reaction was complete, the reaction solution was diluted with saturated aqueous NaCl (100 mL) and extracted three times with dichloromethane (100 mL). The organic phases were combined and washed with anhydrous Na2 SO 4 The mixture was dried at 40° C., filtered by suction, and the solvent was removed by rotary evaporation under reduced pressure. The residue was concentrated to give a crude product as a pale yellow oil. The crude product was purified by column chromatography (DCM:MeOH=100:1 to 50:1) to give 10.5 g of compound 11; LC-MS m / z (M+H) + :952.3. Compound 13 (1.0 g, 1.05 mmol) and DMF (15 mL) were added to a 50 mL single-neck flask at room temperature, followed by the addition of MMAE (759 mg, 1.05 mmol), HOBt (319 mg, 2.3 mmol) and DIPEA (408 μL, 2.3 mmol) in sequence, and the reaction was allowed to stand overnight at room temperature. After the reaction was completed, the reaction solution was directly concentrated under reduced pressure using an oil pump, and the residue was purified by preparative high performance liquid chromatography to give 1.22 g of compound 14 as a white solid; TOF-MS m / z (M+H) + :1530.9. Compound 14 (1 g, 0.653 mmol) and dichloromethane (5 mL) were added to a 25 mL single-neck flask at room temperature, followed by trifluoroacetic acid (2.5 mL), and the reaction was left at room temperature for 6 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure in a water bath, and the residue was purified by preparative high performance liquid chromatography to give 0.76 g of compound 15 as a white solid; TOF-MS m / z (M+Na) + :1496.8.

[0067] Example 6: Synthesis of compound P1-A [ka] Compound 12 (30 mg, 0.02 mmol) and tetrahydrofuran (5 mL) were added to a 25 mL one-neck flask at room temperature, and azido-β-cyclodextrin (24.2 mg, 0.02 mmol), cuprous iodide (2.0 mg, 0.1 mmol) and TEA (4 μL, 0.029 mmol) were added in sequence. The reaction system was heated to 50 °C and reacted for 10 hours. After the reaction was completed, the reaction solution was diluted with 10 mL of ethyl acetate, the organic phase was washed twice with 10 mL of saturated aqueous ammonium chloride solution, the aqueous phase was extracted three times with 10 mL of ethyl acetate, the organic layers were combined, and the mixture was washed with anhydrous Na 2 SO 4 The filtrate was concentrated under reduced pressure, and the residue was purified by preparative high performance liquid chromatography to give 32 mg of compound 16 as a white solid; TOF-MS m / z (M+H) + :2671.4. Compound 16 (30 mg, 11.2 umol) and DMF (1 mL) were added to a 5 mL single-neck flask at room temperature, followed by diethylamine (12 μL, 0.12 mmol), and the reaction was left at room temperature for 12 hours. The DMF solvent was removed under reduced pressure using an oil pump, and the resulting crude yellow oil was sent directly to the reaction without further purification.

[0068] To the above 5 mL one-neck flask, dichloromethane (1 mL) was added at room temperature, followed by compound 6 (45 mg, 112 umol) and carbonyldiimidazole (CDI) (19.9 mg, 0.12 mmol) in sequence, and the reaction was allowed to stand at room temperature for 10 hours. The resulting yellow reaction solution was directly purified by preparative high performance liquid chromatography to obtain 28 mg of compound 17 as a yellowish solid; TOF-MS m / z (M+H) + :2829.3. Compound 17 (25 mg, 8.84 umol) and dichloromethane (4 mL) were added to a 5 mL one-neck flask at room temperature, followed by trifluoroacetic acid (2 mL), and the reaction was allowed to stand at room temperature for 2 hours. After the reaction was completed, the reaction solution was directly concentrated under reduced pressure using a water pump. The residue was purified by preparative high performance liquid chromatography to obtain 13 mg of compound P1-A as a white solid; TOF-MS m / z (M+Na)+ :2695.3.

[0069] Example 7: Synthesis of compound P1-B [ka] Compound mono-6-O-(p-toluenesulfonyl)-β-cyclodextrin (45 mg, 0.035 mmol), 1-amino-11-azido-3,6,9-trioxaundecane (9.0 mg, 0.04 mmol) and 5 mL of DMF were added to a 25 mL one-necked reaction flask, followed by DIPEA (13 μL, 0.07 mmol), and the resulting yellow solution was reacted at room temperature for about 3-4 hours. The reaction solution was purified by preparative high performance liquid chromatography to obtain 42.3 mg of compound 18 as a white solid. LC-MS m / z (M+H) + :1335.6. The subsequent reaction was carried out according to the synthetic route of Example 6 to prepare 9 mg of compound P1-B; TOF-MS m / z (M+Na) + :2870.5.

[0070] Example 8: Synthesis of Compound P1-C [ka] Compound mono-6-O-(p-toluenesulfonyl)-β-cyclodextrin (90 mg, 0.07 mmol), N-tert-butyloxycarbonyl-1,2-ethylenediamine (13.2 mg, 0.08 mmol) and 5 mL of DMF were added to a 25 mL one-necked reaction flask, followed by DIPEA (26 μL, 0.14 mmol), and the resulting yellow solution was reacted at room temperature for about 4 hours. The reaction solution was purified by preparative high performance liquid chromatography to obtain 77 mg of compound 20 as a white solid; LC-MS m / z (M+H) + :1177.5. Compound 20 (70 mg, 0.06 mmol) and dichloromethane (4 mL) were added to a 5 mL one-neck flask at room temperature, followed by trifluoroacetic acid (2 mL), and the reaction was left at room temperature for 4 hours. After the reaction was complete, trifluoroacetic acid and dichloromethane were removed by rotary evaporation under reduced pressure, and the residue was sent directly to the next reaction. In a 25 mL single-neck flask, the above crude compound (0.06 mmol based on the starting material loading), PyBOP (63.7 mg, 0.12 mmol), compound 15 (88 mg, 0.06 mmol) and DMF (5 mL) were added, and then the resulting solution was cooled to 0 °C under ice-water bath conditions. DIPEA (22 μL, 0.12 mmol) was added dropwise to the reaction flask, and the resulting reaction solution was allowed to slowly come to room temperature naturally and react overnight. The resulting yellow reaction solution was directly purified by preparative high performance liquid chromatography to obtain 55 mg of compound 21; TOF-MS m / z (M+Na) + :2655.2.

[0071] Compound 21 (50 mg, 19 umol) and DMF (1 mL) were added to a 5 mL single-neck flask at room temperature, followed by diethylamine (35 μL, 0.19 mmol), and the reaction was left at room temperature overnight. The DMF solvent was removed under reduced pressure using an oil pump, and the resulting crude yellow oil was sent directly to the reaction without further purification. To the above 5 mL one-neck flask, dichloromethane (1 mL) was added at room temperature, followed by compound 6 (77 mg, 190 umol) and carbonyldiimidazole (CDI) (32 mg, 0.19 mmol) in sequence, and the reaction was allowed to stand at room temperature for 10 hours. The resulting yellow reaction solution was directly purified by preparative high performance liquid chromatography to obtain 46 mg of compound 22 as a yellowish solid; TOF-MS m / z (M+H) + :2813.5. Compound 22 (45 mg) and dichloromethane (4 mL) were added to a 5 mL one-neck flask at room temperature, followed by trifluoroacetic acid (2 mL), and the reaction was left at room temperature for 2 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure in a water bath. The residue was purified by preparative high performance liquid chromatography to give 20 mg of compound P1-C as a white solid; TOF-MS m / z (M+Na) + :2657.2.

[0072] Example 9: Synthesis of compound P1-D [ka] Compound 18 (40 mg) and DMF (5 mL) were added to a 25 mL single-neck flask at room temperature, followed by Pd / C (40 mg), and the atmosphere of the system was replaced with hydrogen three times. The reaction was left at room temperature for 1 h. The reaction solution was filtered to remove the catalyst. The DMF solvent was removed under reduced pressure by oil pump, and the resulting yellow crude oil 23 was sent directly to the reaction without further purification. Following the synthetic route of Example 8, 9.9 mg of yellowish solid compound P1-D was prepared; TOF-MS m / z (M+Na) + :2789.4.

[0073] Example 10: Synthesis of compound P1-E [ka] Compound 10 (100 mg) and DMF (10 mL) were added to a 25 mL one-neck flask at room temperature, followed by Pd / C (100 mg), and the atmosphere of the system was replaced with hydrogen three times. The reaction was left at room temperature for 3 hours. The reaction solution was filtered to remove the catalyst. The DMF solvent was removed under reduced pressure by oil pump. The obtained yellow crude oil 24 was sent directly to the reaction without further purification. Following the synthetic route of Example 8, 23 mg of yellowish solid compound P1-E was prepared; TOF-MS m / z (M+Na) + :2012.5.

[0074] Example 11: Synthesis of Compound P1-F [ka] Compound maltobionic acid (200 mg, 0.56 mmol), N-tert-butyloxycarbonyl-1,2-ethylenediamine (196 mg, 1.2 mmol) and 10 mL of DMF were added to a 25 mL one-neck reaction flask, followed by DIPEA (0.22 mL, 1.2 mmol), and the resulting yellow solution was reacted at room temperature for about 4 hours. The reaction solution was purified by preparative high performance liquid chromatography to obtain a white solid compound. The above compound and dichloromethane (6 mL) were added to a 25 mL one-neck flask at room temperature, followed by trifluoroacetic acid (3 mL), and the reaction was left at room temperature for 2 hours. After the reaction was completed, rotary evaporation was performed under reduced pressure to remove trifluoroacetic acid and dichloromethane, which was sent directly to the next reaction; LC-MS m / z (M+H) + :401.1. 23 mg of yellowish solid compound P1-F was prepared according to the synthetic route of Example 8. ;TOF-MS m / z(M+H) + :1858.1.

[0075] Example 12: Synthesis of compound P1-G [ka] Referring to the synthesis route of Example 6, 15 mg of white solid compound P1-G was obtained from compound 10 and compound 12; TOF-MS m / z (M+H) + :2071.4.

[0076] Example 13: Synthesis of compound P1-H [ka] 12 mg of white solid compound P1-H was prepared according to the synthetic route of Example 6; TOF-MS m / z (M+H) + :2291.6.

[0077] Example 14: Synthesis of compound P1-I [ka] Compound 27 was prepared according to the synthetic route of Example 3; LC-MS m / z (M+H) + :455.4. Following the synthetic route of Example 6, 11 mg of white solid compound P1-I was prepared; TOF-MS m / z (M+H) + :1989.2.

[0078] Example 15: Synthesis of Compound P1-J [ka] Maltose (500 mg, 1.46 mmol), 1,1-dimethylethyl 13-amino-5,8,11-trioxa-2-azatridecanoate (436 mg, 1.46 mmol) and 15 mL of absolute ethanol were added to a 25 mL one-necked reaction flask, and the reaction was left at room temperature for about 4 hours. The reaction flask was cooled in an ice-water bath, and sodium cyanoborohydride (186 mg, 2.92 mmol) was added twice, and the reaction was allowed to proceed at a constant temperature for 1 hour. The reaction solution was purified by preparative high performance liquid chromatography, and then freeze-dried to obtain a white solid compound. The above compound and dichloromethane (10 mL) were added to a 25 mL one-necked flask at room temperature, followed by trifluoroacetic acid (5 mL), and the reaction was allowed to proceed at room temperature for 3 hours. After the reaction was completed, rotary evaporation was performed under reduced pressure to remove trifluoroacetic acid and dichloromethane, and the residue was sent directly to the next reaction; LC-MS m / z (M+H) + :519.3. Following the synthetic route of Example 8, 8 mg of white solid compound P1-J was prepared; TOF-MS m / z (M+Na) + :1998.2.

[0079] Example 16: Synthesis of compound P1-K [ka] 21 mg of white solid compound P1-K was prepared according to the synthetic route of Example 8 and Example 14; TOF-MS m / z (M+Na) + :1866.0.

[0080] Example 17: Synthesis of compound P1-L [ka] In a 25mL one-neck flask, 1-amino-11-azido-3,6,9-trioxaundecane (60mg, 0.27mmol) and tetrahydrofuran (5mL) were added at room temperature, followed by N-Boc-aminopropyne (44mg, 0.27mmol), cuprous iodide (2.0mg, 0.1mmol) and TEA (40μL, 0.29mmol) in sequence. The mixture was heated to 50℃ and reacted for 10 hours, then filtered, and the residue was purified by preparative high performance liquid chromatography to obtain 82mg of compound 29 as a white solid; LC-MS m / z(M+H) + :374.3. Compound 30 was prepared according to the synthetic route of Example 14; LC-MS m / z (M+H) + :600.5. Following the synthetic route of Example 8, 17 mg of white solid compound P1-L was prepared; TOF-MS m / z (M+Na) + :2079.1.

[0081] Example 18: Synthesis of compound P1-M [ka] 21 mg of white solid compound P1-M was prepared according to the synthetic route of Example 16; TOF-MS m / z (M+Na) + :2299.1.

[0082] Example 19: Synthesis of Compound P1-N [ka] 13 mg of solid compound P1-N was prepared according to the synthetic route of Example 16; TOF-MS m / z (M+H)+ :1953.2.

[0083] Example 20: Synthesis of Compounds P1-O8 [ka] Maltose (50 mg, 0.146 mmol), N-Boc-ethylenediamine (24 mg, 0.146 mmol) and 5 mL of absolute ethanol were added to a 25 mL single-necked reaction flask, and the reaction was left at room temperature for about 4 hours. The reaction flask was cooled in an ice-water bath, and sodium cyanoborohydride (18.6 mg, 0.292 mmol) was added twice and allowed to react at room temperature for 1 hour. The reaction solution was purified by preparative high performance liquid chromatography, and then freeze-dried to obtain a solid compound. The above compound and dichloromethane (4 mL) were added to a 25 mL single-necked flask at room temperature, followed by trifluoroacetic acid (2 mL), and the reaction was left at room temperature for 3 hours. After the reaction was complete, rotary evaporation was performed under reduced pressure to remove trifluoroacetic acid and dichloromethane to obtain crude compound 31, which was sent directly to the next reaction without purification. LC-MS m / z (M+H) + :387.2. In a 25 mL one-neck flask, the above crude compound (0.146 mmol based on the starting material loading), N-tert-butyloxycarbonyl-heptaethylene glycol-carboxylic acid (81 mg, 0.146 mmol), PyBOP (116 mg, 0.219 mmol) and DMF (5 mL) were added, and the resulting solution was then cooled to 0° C. under ice-water bath conditions. DIPEA (40 μL, 0.219 mmol) was added dropwise to the reaction flask, and the resulting solution was allowed to slowly come to room temperature and react overnight. The resulting yellow reaction solution was directly concentrated under reduced pressure by oil pump to obtain a yellow oil. To the above 25 mL one-neck flask containing the oil, dichloromethane (4 mL) was added at room temperature, followed by trifluoroacetic acid (2 mL), and the reaction was left at room temperature for 2 hours. After the reaction was completed, rotary evaporation was performed under reduced pressure to remove trifluoroacetic acid and dichloromethane to obtain the crude product. The crude product was purified by preparative high performance liquid chromatography column and lyophilized to give 66 mg of compound 32 as a white solid; LC-MS m / z (M+H) + :810.9. 15 mg of solid compound P1-O8 was prepared according to the synthetic route of Example 8; TOF-MS m / z (M+Na) + :2289.2.

[0084] Example 21: Synthesis of Compounds P1-P8 [ka] Compound 33 was prepared according to the synthetic route of Example 19; LC-MS m / z (M+H) + :836.9. 23 mg of solid compounds P1-P8 were prepared according to the synthetic route of Example 6; TOF-MS m / z (M+Na) + :2289.2.

[0085] Example 22: Synthesis of Compound P1-Q [ka] Compound 34 was prepared according to the synthetic route of Example 19; LC-MS m / z (M+H)+ :512.5 Following the synthetic route of Example 6, 9 mg of solid compound P1-Q was prepared; TOF-MS m / z (M+H) + :2024.1.

[0086] Example 23: Synthesis of compound P1-R [ka] Compound 35 was prepared according to the synthetic route of Example 19; LC-MS m / z (M+H) + :486.5 14 mg of solid compound P1-R was prepared according to the synthetic route of Example 8; TOF-MS m / z (M+H) + :1943.2.

[0087] Example 24: Synthesis of compound P1-S [ka] 24 mg of solid compound P1-S was prepared according to the synthetic route of Example 6; TOF-MS m / z (M+Na) + :1929.1.

[0088] Example 25: Synthesis of compound P1-T [ka] Following the synthetic route of Example 6, 16 mg of solid compound P1-T was prepared; TOF-MS m / z (M+H) + :1803.5.

[0089] Example 26: Synthesis of compound P1-U [ka] 12 mg of solid compound P1-U was prepared according to the synthetic route of Example 8; TOF-MS m / z (M+H) + :1693.9.

[0090] Example 27: Synthesis of Compound P1-V [ka] Following the synthetic route of Example 8, 19 mg of solid compound P1-V was prepared; TOF-MS m / z (M+H) + :1840.1.

[0091] Example 28: Synthesis of compound P1-W [ka] 32 mg of solid compound P1-W was prepared according to the synthetic route of Example 6; TOF-MS m / z (M+H) + :2127.1291.

[0092] Example 29: Synthesis of Compounds P1-X2 [ka] 27 mg of solid compound P1-W was prepared according to the synthetic route of Example 6; TOF-MS m / z (M+Na) + :2853.2939.

[0093] Example 30: Synthesis of Compound P1-Y8 [ka] Step 1: Compound NH 2 -PEG 8 Synthesis of -β-CD In a 25 mL single-neck flask, compound 27-amino-4,7,10,13,16,19,22,25-octaoxaheptadecanoic acid (100 mg, 0.22 mmol) and DMF (5 mL) were added at room temperature, followed by HOSu (28 mg, 0.23 mmol) and DCC (48 mg, 0.23 mmol), and the reaction was left at room temperature for 3 h. The DMF solvent was removed under reduced pressure with an oil pump. The resulting crude yellow oil was dissolved in 10 mL of DCM and filtered to remove insoluble solids; the filter cake was washed three times with DCM; the organic liquids were combined and spun dry in a 45 °C water bath with a water pump, and the residue was sent directly to the next reaction. The crude compound (0.22 mmol) and DMF (5 mL) were added to a 25 mL single-neck flask at room temperature, followed by mono(6-amino-6-deoxy)-β-cyclodextrin (275 mg, 0.23 mmol) and DIPEA (74 μL, 0.44 mmol), and the reaction was allowed to stand at room temperature for 2 hours. After HPLC monitoring showed the reaction was complete, the reaction solution was purified by preparative high performance liquid chromatography to obtain a purified solution. The purified solution was lyophilized to obtain compound NH. 2 -PEG 8 -β-CD (145 mg) was obtained; LC-MS m / z (M+H) + :1157.7. Step 2: Synthesis of Compound P1-Y8 Compound 15 and NH 2 -PEG 8 Starting from -β-CD, 13 mg of solid compound P1-Y8 was prepared according to the synthesis route of Example 8; TOF-MS m / z (M+H) + :3014.4188.

[0094] Example 31: Synthesis of Compound P1-Z [ka] Step 1: Synthesis of compound 36 Azide-PEG8-NHS (100 mg, 0.17 mmol) and DMF (5 mL) were added to a 25 mL single-neck flask at room temperature, followed by meglumine (34 mg, 0.17 mmol) and DIPEA (66 μL, 0.4 mmol), and the reaction was left at room temperature for 2 h. After HPLC monitoring showed the reaction was complete, the reaction solution was purified by preparative high performance liquid chromatography to obtain a purified solution. The purified solution was lyophilized to obtain compound 36 (120 mg); LC-MS m / z (M+H) + :645.7. Step 2: Synthesis of compound P1-Z Starting from compound 12 and compound 36, 18 mg of solid compound P1-Z was prepared according to the synthesis route of Example 6; TOF-MS m / z (M+H) + :2157.1796.

[0095] Example 32: Preparation of Compound 37 [ka] In a 1L single-neck flask, 5-tert-butyl Boc-L-glutamate (50g, 164.8mmol, 1.0eq) was completely dissolved in 250mL of THF, and propargylamine (9.97g, 181.3mmol, 1.1eq), HOBt (22.25g, 164.8mmol, 1.0eq), DIEA (54.3mL, 329.6mmol, 2.0eq) and EDCI (37.7g, 197.7mmol, 1.2eq) were added sequentially under ice-water bath conditions. The reaction was allowed to react at room temperature for 2 hours. After TLC monitoring showed that the reaction was complete, the reaction solution was poured into 600mL of ice water and extracted with ethyl acetate three times (200mL x 3); the organic liquid was combined and washed twice with saturated salt solution (300mL x 2) and diluted with anhydrous NaCl. 2 SO 4 The mixture was dried at 40° C., filtered and concentrated to give the crude product, which was purified with petroleum ether / ethyl acetate (2 / 1) to give 35.5 g of compound 37a; LC-MS m / z (M+H) + :341.2. In a 1L single-neck flask, compound 37a (20 g) was completely dissolved in 150 mL of dichloromethane, then 150 mL of trifluoroacetic acid was added, and the reaction was left at room temperature for 1 h. After TLC monitoring showed that the reaction was complete, the reaction solution was concentrated at 45° C. with a water pump to obtain an oily crude product. The crude product was added to methyl tert-ether to crystallize overnight. The reaction solution was filtered, and the filter cake was dried by blowing air at 45° C. for 4 h to obtain 7.68 g of product; LC-MS m / z (M+H) + :184.2.

[0096] Example 33: Preparation of Compound 39 [ka] In a 1 L single-neck flask, 5-tert-butyl Boc-L-glutamate (50 g, 164.8 mmol, 1.0 equiv.) was completely dissolved in 300 mL of THF. FmocNH-(CH 2 ) 2 -NH 2 (51.1 g, 181.3 mmol, 1.1 equiv), HOBt (22.25 g, 164.8 mmol, 1.0 equiv), DIEA (54.3 mL, 329.6 mmol, 2.0 equiv) and EDCI (37.7 g, 197.7 mmol, 1.2 equiv) were added sequentially under ice-water bath conditions, and the reaction was allowed to react at room temperature for 2 h. After TLC monitoring showed that the reaction was complete, the reaction solution was poured into 600 mL of ice water and extracted three times with ethyl acetate (300 mL x 3); the organic liquid was combined, washed twice with saturated salt solution (300 mL x 2), and extracted with anhydrous NaCl. 2 SO 4 The mixture was dried at 40° C., filtered and concentrated to give the crude product. The crude product was purified with petroleum ether / ethyl acetate (2 / 1) to give 57.5 g of pure product; LC-MS m / z (M+H) + :567.2. In a 1L single-neck flask, compound 38 (20 g) was completely dissolved in 150 mL of dichloromethane, then 150 mL of trifluoroacetic acid was added, and the reaction was left at room temperature for 1 hour. After TLC monitoring showed that the reaction was complete, the reaction solution was concentrated at 45° C. to give a crude oil. The crude product was crystallized in methyl tert-ether overnight and filtered. The filter cake was dried by blowing air at 45° C. for 4 hours and weighed to give 11.7 g of product; LC-MS m / z (M+H) + :412.2.

[0097] Example 34: Preparation of Compound 40 [ka] In a 50 mL single-neck flask, compound 37 (5.0 g, 27.1 mmol) and compound 6 (10.8 g, 27.1 mmol) were mixed and completely dissolved in 20 mL of DMF, and then EEDQ (13.4 g, 54.2 mmol) was added under ice-water bath conditions. After the addition was completed, the reaction was left at room temperature for 2 hours. After HPLC monitoring showed that the reaction was complete, the reaction solution was purified by preparative high performance liquid chromatography and lyophilized to give 7.84 g of product; LC-MS m / z (M+H) + :465.2.

[0098] Example 35: Preparation of Compound 44 [ka] In a 50mL one-neck flask, compound 41 (1.0g, 1.51mmol, see patent application WO2020063676A1) was completely dissolved in 15mL of DMF, 5% Pd / C (1.0g) was added, and the hydrogenation reaction was carried out at room temperature for 2 hours. After the reaction was found to be complete by HPLC monitoring, the filtrate was obtained by filtration and designated as filtrate 1; In a 25mL one-neck flask, compound 40 (0.85g, 1.51mmol) was completely dissolved in 15mL DMF, then DCC (1.55g, 7.56mmol) and pentafluorophenol (0.31g, 1.66mmol) were added under ice-water bath conditions, and the reaction was allowed to stand at room temperature for 2 hours. After the reaction was found to be complete by TLC monitoring, the filtrate was obtained by filtration, designated as filtrate 2; DIEA (0.37 mL, 2.26 mmol) was added to filtrate 1 under ice-water bath conditions, followed by filtrate 2, and the reaction was left at room temperature for 1 h. After HPLC monitoring showed the reaction was complete, the reaction solution was purified by preparative high performance liquid chromatography to obtain the prepared solution. The prepared solution was lyophilized to obtain 502 mg of compound 44 as a white solid; LC-MS m / z (M+H) + :985.2.

[0099] Example 36: Preparation of Compound 45 [ka] In a 50 mL single-neck flask, compound 44 (500 mg, 0.51 mmol) was completely dissolved in 10 mL of DMF, then PyBOP (396 mg, 0.76 mmol), HOBt (103 mg, 0.76 mmol) and exatecan mesylate (270 mg, 0.51 mmol) were added sequentially under ice-water bath conditions, followed by DIEA (252 μL, 1.52 mmol) and the reaction was left at room temperature for 2 hours. After HPLC monitoring showed the reaction was complete, the reaction solution was purified by preparative high performance liquid chromatography to obtain the prepared solution. The prepared solution was lyophilized to obtain 500 mg of compound 45 as a white solid; LC-MS m / z (M+H) + :1402.2.

[0100] Example 37: Synthesis of compound P2-A [ka] In a 25 mL one-neck flask, compound 45 (50 mg, 0.035 mmol) and β-cyclodextrin-N 3(41 mg, 0.035 mmol) was completely dissolved in 5 mL of DMF, and then cuprous iodide (2 mg, 0.007 mmol) and TEA (10 μL, 0.07 mmol) were added. The resulting solution was heated to 50° C. and reacted for 10 hours, and the reaction process was monitored by HPLC. After the reaction was completed, the reaction solution was purified by preparative high performance liquid chromatography to obtain a preparation solution of the product. The preparation solution was freeze-dried to obtain 59 mg of compound 46; TOF-MS m / z (M+Na) + :2583.0. In a 25mL one-neck flask, compound 46 (50mg, 0.019mmol) was completely dissolved in 5mL of nitromethane, then zinc bromide (88mg, 0.39mmol) was added, and the reaction was left at room temperature for 1h. The reaction process was monitored by HPLC. After the reaction was completed, the reaction solution was concentrated under reduced pressure by water pump at 45℃ to obtain the crude product. The crude product was purified by preparative high performance liquid chromatography to obtain the product preparation solution. The preparation solution was freeze-dried and weighed to obtain 30mg of compound P2-A; TOF-MS m / z(M+H) + :2404.9.

[0101] Example 38: Synthesis of compound P2-B [ka] 24 mg of compound P1-B was prepared according to the synthetic route of Example 37; TOF-MS m / z (M+H) + :2580.0.

[0102] Example 39: Synthesis of Compound 49 [ka] In a 50 mL single-neck flask, compound 39 (5.0 g, 12.1 mmol) and compound 6 (4.83 g, 12.1 mmol) were mixed and completely dissolved in 25 mL of DMF, and then EEDQ (6.0 g, 24.3 mmol) was added under ice-water bath conditions. After the addition was completed, the reaction was left at room temperature for 2 hours. After HPLC monitoring showed that the reaction was complete, the reaction solution was purified by preparative high performance liquid chromatography and then lyophilized to give 5.1 g of product; LC-MS m / z (M+H) + :792.3. In a 50 mL single-neck flask, compound 47 (0.3 g, 0.378 mmol) was completely dissolved in 5 mL of DMF, then 0.3 mL of diethylamine was added, and the resulting solution was stirred at room temperature for 1 h. After TLC monitoring showed that the reaction was complete, the reaction solution was concentrated under reduced pressure to remove DMF and diethylamine to obtain the crude product, which was directly sent to the next reaction. The crude product was dissolved in 5 mL of DMF, after which mono-6-O-(p-toluenesulfonyl)-β-cyclodextrin (488 mg, 0.378 mmol) was added and the reaction was immersed in N 2 After the reaction was found to be complete by HPLC monitoring, the reaction solution was purified by preparative high performance liquid chromatography and lyophilized to give 601 mg of compound 49; TOF-MS m / z (M+H) + :1686.9.

[0103] Example 40: Synthesis of compound P2-C [ka] In a 25mL single-neck flask, compound 49 (1.05g, 0.62mmol) was completely dissolved in 10mL DMF. DCC (653mg, 3.1mmol) and pentafluorophenol (119mg, 0.63mmol) were added under ice-water bath conditions, and the reaction was allowed to stand at room temperature for 2 hours. After TLC monitoring showed that the reaction was complete, the reaction solution was filtered to obtain the filtrate; DIEA (0.22 mL, 1.24 mmol) was added to the above filtrate under ice-water bath condition, followed by compound 42 (272 mg, 0.62 mmol) and the reaction was left at room temperature for 1 h. After the reaction was found to be complete by HPLC monitoring, the reaction solution was purified by preparative high performance liquid chromatography to obtain the prepared solution. The prepared solution was lyophilized to obtain 588 mg of compound 51 as a white solid; TOF-MS m / z (M+H) + :2105.8. In a 50 mL single-neck flask, compound 51 (500 mg, 0.24 mmol) was completely dissolved in 10 mL of DMF, then PyBOP (188 mg, 0.36 mmol), HOBt (49 mg, 0.36 mmol) and exatecan mesylate (140 mg, 0.26 mmol) were added sequentially under ice-water bath conditions, followed by DIEA (86 μL, 0.48 mmol) and the reaction was left at room temperature for 2 h. After HPLC monitoring showed the reaction was complete, the reaction solution was purified by preparative high performance liquid chromatography to obtain the prepared solution. The prepared solution was lyophilized to obtain 403 mg of compound 52 as a solid; TOF-MS m / z (M+Na) + :2544.0. In a 25 mL single-neck flask, compound 52 (60 mg, 0.024 mmol) was completely dissolved in 5 mL of nitromethane, then zinc bromide (109 mg, 0.48 mmol) was added and the reaction was left at room temperature for 1 h. After HPLC monitoring showed the reaction was complete, the reaction solution was concentrated under reduced pressure with a water pump at 45 °C to obtain the crude product. The crude product was purified by preparative high performance liquid chromatography to obtain a preparation solution of the product. The preparation solution was lyophilized and weighed to obtain 29 mg of compound P2-C; TOF-MS m / z (M+H) + :2365.9.

[0104] Example 41: Synthesis of compound P2-D [ka] 24 mg of compound P2-D was prepared according to the synthetic route of Examples 39 and 40; TOF-MS m / z (M+H) +:2500.3.

[0105] Example 42: Synthesis of compound P2-E [ka] Compound 53 was prepared according to the synthetic route of Example 29; Compound 54 was prepared according to the synthetic route of Example 31; 33 mg of compound P2-E was prepared according to the synthetic route of Example 40; TOF-MS m / z (M+Na) + :1244.8.

[0106] Example 43: Synthesis of compound P2-F [ka] 16 mg of compound P2-F was prepared according to the synthetic route of Examples 39 and 40; TOF-MS m / z (M+H) + :1590.7.

[0107] Example 44: Synthesis of compound P2-G [ka] In a 25mL one-neck flask, compound 45 (50mg, 0.035mmol) and compound 10 (20mg, 0.035mmol) were mixed and completely dissolved in 5mL of DMF, and then cuprous iodide (2mg, 0.007mmol) and TEA (10μL, 0.07mmol) were added, and the resulting solution was heated to 50℃ and reacted for 10 hours. The reaction process was monitored by HPLC. After the reaction was completed, the reaction solution was purified by preparative high-performance preparation liquid phase chromatography to obtain a preparation solution of the product. The preparation solution was freeze-dried and weighed to obtain 43mg of the product; TOF-MS m / z(M+Na) + :1981.9. In a 25mL one-neck flask, compound 55 (40mg, 0.02mmol) was completely dissolved in 3mL of nitromethane, then zinc bromide (92mg, 0.4mmol, 20eq) was added, and the reaction was left at room temperature for 1h. The reaction process was monitored by HPLC. After the reaction was completed, the reaction solution was concentrated under reduced pressure by water pump at 45℃ to obtain the crude product. The crude product was purified by preparative high performance liquid chromatography to obtain the product preparation solution. The preparation solution was freeze-dried and weighed to obtain 25mg of product P2-G; TOF-MS m / z(M+H) + :1803.7.

[0108] Example 45: Synthesis of compound P2-H [ka] 16 mg of compound P2-H was prepared according to the synthetic route of Example 44; TOF-MS m / z (M+H) + :2023.9.

[0109] Example 46: Synthesis of compound P2-I [ka] In a 25mL one-neck flask, compound 45 (50mg, 0.035mmol) and compound 27 (16mg, 0.035mmol) were mixed and completely dissolved in 5mL of DMF, and then cuprous iodide (2mg, 0.007mmol) and TEA (10μL, 0.07mmol) were added, and the resulting solution was heated to 50℃ and reacted for 10 hours. The reaction process was monitored by HPLC. After the reaction was completed, the reaction solution was purified by preparative high performance liquid chromatography to obtain a preparation solution of the product. The preparation solution was freeze-dried and weighed to obtain 43mg of the product; TOF-MS m / z(M+H) + :1955.9. In a 25mL one-neck flask, compound 56 (40mg, 0.021mmol) was completely dissolved in 5mL of nitromethane, then zinc bromide (97mg, 0.42mmol) was added, and the reaction was left at room temperature for 1h. The reaction process was monitored by HPLC. After the reaction was completed, the reaction solution was concentrated under reduced pressure by water pump at 45℃ to obtain the crude product. The crude product was purified by preparative high performance liquid chromatography to obtain the product preparation solution. The preparation solution was freeze-dried and weighed to obtain 25mg of compound P2-I; TOF-MS m / z(M+H) + :1699.7.

[0110] Example 47: Synthesis of compound P2-J [ka] In a 50 mL single-neck flask, compound 47 (3.0 g, 3.78 mmol) was completely dissolved in 15 mL of DMF, then 3 mL of diethylamine was added, and the resulting solution was stirred at room temperature for 1 h. After TLC monitoring showed that the reaction was complete, the reaction solution was concentrated under reduced pressure to remove DMF and diethylamine to obtain the crude product. The crude product was dissolved in 20 mL of methanol, then maltose (1.29 g, 3.78 mmol) was added, and the reaction was immersed in N 2 The mixture was left at room temperature for 2 hours in the presence of NaBH 4 (285 mg, 7.56 mmol) was added in one portion under ice-water bath conditions and allowed to react for 30 minutes. After TLC monitoring showed the reaction was complete, 20 mL of water was added, and the resulting reaction solution was extracted three times with ethyl acetate (30 mL x 3), the organic liquids were combined, washed three times with saturated NaCl solution, and then extracted with anhydrous Na 2 SO 4 The mixture was dried at 40° C., filtered, and the filtrate was concentrated to give the crude product, which was purified by preparative high performance liquid chromatography, lyophilized, and weighed to give 1.4 g of solid; LC-MS m / z (M+H) + :896.3. In a 25 mL single-neck flask, compound 57 (677 mg, 0.75 mmol) was completely dissolved in 15 mL of DMF, DCC (780 mg, 3.78 mmol) and pentafluorophenol (153 mg, 0.83 mmol) were added under ice-water bath conditions, and the reaction was left at room temperature for 2 hours. After TLC monitoring showed that the reaction was complete, the reaction solution was filtered to obtain a filtrate; DIEA (187 μL, 1.13 mmol) was added to the above filtrate under ice-water bath conditions, and then compound 42 (0.75 mmol) was added, and the reaction was left at room temperature for 1 hour. After HPLC monitoring showed that the reaction was complete, the reaction solution was purified by preparative high performance liquid chromatography to obtain a preparation solution. The preparation solution was freeze-dried to obtain 350 mg of compound 59 as a white solid; LC-MS m / z (M+H) + :1315.6.

[0111] In a 50 mL single-neck flask, compound 59 (200 mg, 0.15 mmol) was completely dissolved in 10 mL of DMF, and PyBOP (118 mg, 0.22 mmol), HOBt (31 mg, 0.22 mmol), and exatecan mesylate (80 mg, 0.15 mmol) were added sequentially under ice-water bath conditions, followed by DIEA (75 μL, 0.45 mmol), and the reaction was left at room temperature for 2 h. After HPLC monitoring showed the reaction was complete, the reaction solution was purified by preparative high performance liquid chromatography to obtain the prepared solution. The prepared solution was lyophilized to obtain 150 mg of compound 60 as a white solid; TOF-MS m / z (M+H) + :1732.7. In a 25mL one-neck flask, compound 60 (50mg, 0.028mmol) was completely dissolved in 5mL of nitromethane, then zinc bromide (130mg, 0.57mmol) was added, and the reaction was left at room temperature for 1h. The reaction process was monitored by HPLC. After the reaction was completed, the reaction solution was concentrated under reduced pressure by water pump at 45℃ to obtain the crude product. The crude product was purified by preparative high performance liquid chromatography to obtain the product preparation solution. The preparation solution was freeze-dried and weighed to obtain 30mg of product; TOF-MS m / z(M+H)+ :1576.6.

[0112] Example 48: Synthesis of compound P2-K [ka] 28 mg of compound P2-K was prepared according to the synthetic route of Example 47; TOF-MS m / z (M+H) + :1708.7.

[0113] Example 49: Synthesis of compound P2-L [ka] 33 mg of compound P2-L was prepared according to the synthetic route of Example 47; TOF-MS m / z (M+H) + :1789.8.

[0114] Example 50: Synthesis of compound P2-M [ka] Following the synthetic route of compound 30, 77 mg of compound 61 was prepared; LC-MS m / z (M+H) + :820.9. 15 mg of compound P2-M was prepared according to the synthetic route of Example 40; TOF-MS m / z (M+H) + :2010.0.

[0115] Example 51: Synthesis of Compound P2-N [ka] Following the synthetic route of compound 30, 62 mg of compound 64 was prepared; LC-MS m / z (M+H) + :496.5. 23 mg of compound P2-N was prepared according to the synthetic route of Example 40; TOF-MS m / z (M+Na) + :1707.7.

[0116] Example 52: Synthesis of compound P2-O [ka] Following the synthetic route of compound 32, 71 mg of compound 66 was prepared; LC-MS m / z (M+H) + :590.6. 34 mg of compound P2-O was prepared according to the synthetic route of Example 40; TOF-MS m / z (M+Na) + :1801.9.

[0117] Example 53: Synthesis of compound P2-P [ka] 20 mg of compound P2-P was prepared according to the synthetic route of Example 46; TOF-MS m / z (M+H) + :2081.1.

[0118] Example 54: Synthesis of Compound P2-Q [ka] 29 mg of compound P2-Q was prepared according to the synthetic route of Example 46; TOF-MS m / z (M+H) + :1756.7.

[0119] Example 55: Synthesis of Compound P2-R [ka] 18 mg of compound P2-R was prepared according to the synthetic route of Example 40; TOF-MS m / z (M+H) + :1675.9.

[0120] Example 56: Synthesis of compound P2-S [ka] 33 mg of compound P2-S was prepared according to the synthetic route of Example 40; TOF-MS m / z (M+H) + :1558.9.

[0121] Example 57: Synthesis of compound P2-T [ka] 42 mg of compound P2-T was prepared according to the synthetic route of Example 40; TOF-MS m / z (M+H) + :1426.8.

[0122] Example 58: Synthesis of compound P2-U [ka] 30 mg of compound P2-U was prepared according to the synthetic route of Example 46; TOF-MS m / z (M+H) + :1640.1.

[0123] Example 59: Synthesis of compound P2-V [ka] 18 mg of compound P2-V was prepared according to the synthetic route of Example 46; TOF-MS m / z (M+H) + :1860.0.

[0124] Example 60: Synthesis of compound P2-W [ka] 21 mg of compound P2-W was prepared according to the synthetic route of Example 46; TOF-MS m / z (M+H) + :1535.8.

[0125] Example 61: Synthesis of Compounds P2-X2 [ka] 11 mg of compound P2-X2 was prepared according to the synthetic route of Example 37; TOF-MS m / z (M+H) + :2563.9238.

[0126] Example 62: Synthesis of Compound P2-Y8 [ka] 12 mg of compound P2-Y8 was prepared according to the synthetic route of Example 40; TOF-MS m / z (M+H) + :2747.0488.

[0127] Example 63: Synthesis of Compound P2-Z [ka] Compound 45 (70 mg, 0.05 mmol) and PJA-PEG8-N3 (32 mg, 0.05 mmol) were mixed and completely dissolved in 5 mL of DMF, and then cuprous iodide (1 mg, 0.005 mmol) and triethylamine (14 μL, 0.1 mmol) were added, and the resulting reaction solution was heated to 50° C. and reacted overnight. The reaction process was monitored by HPLC. After the reaction was completed, the reaction solution was purified by preparative high performance liquid chromatography to obtain a preparation solution of the product. The preparation solution was freeze-dried to obtain 69 mg of the product. In a 25mL one-neck flask, the above compound (69mg) was completely dissolved in 5mL of nitromethane, then zinc bromide (230mg, 1mmol) was added, and the reaction was left at room temperature for 1 hour. The reaction process was monitored by HPLC. After the reaction was completed, the reaction solution was concentrated under reduced pressure by water pump at 45℃ to obtain the crude product. The crude product was purified by preparative high performance liquid chromatography to obtain the preparation solution of the product. The preparation solution was freeze-dried and weighed to obtain 44mg of compound P2-Z; TOF-MS m / z(M+H) + :1889.8077.

[0128] Example 64: Synthesis of compound P-1 [ka] Compound 45 (50 mg, 0.035 mmol) and 1-methoxy-11-azido-3,6,9-trioxane (8.3 mg, 0.035 mmol) were mixed and completely dissolved in 5 mL of DMF, and then cuprous iodide (1.4 mg, 0.007 mmol) and triethylamine (10 μL, 0.07 mmol) were added, and the resulting reaction solution was heated to 50° C. and reacted for 10 hours. The reaction process was monitored by HPLC. After the reaction was completed, the reaction solution was purified by preparative high performance liquid chromatography to obtain a preparation solution of the product. The preparation solution was freeze-dried to obtain 41 mg of the product; TOF-MS m / z (M+H) + :1634.7. In a 25mL one-neck flask, compound 67 (40mg, 0.024mmol) was completely dissolved in 5mL of nitromethane, then zinc bromide (110mg, 0.48mmol) was added, and the reaction was left at room temperature for 1 hour. The reaction process was monitored by HPLC. After the reaction was completed, the reaction solution was concentrated under reduced pressure by water pump at 45℃ to obtain the crude product. The crude product was purified by preparative high performance liquid chromatography to obtain the product preparation solution. The preparation solution was freeze-dried and weighed to obtain 25mg of compound P-1; TOF-MS m / z(M+H) + :1478.6.

[0129] Example 65: Synthesis of Compound P-2 [ka] Compound 45 (50 mg, 0.035 mmol) and 28-azido-2,5,8,11,14,17,20,23,26-nonaoxaoctacosane (16.1 mg, 0.035 mmol) were mixed and completely dissolved in 5 mL of DMF, and then cuprous iodide (1.4 mg, 0.007 mmol) and triethylamine (10 μL, 0.07 mmol) were added. The resulting reaction solution was heated to 50 °C and reacted for 10 hours, and the reaction process was monitored by HPLC. After the reaction was completed, the reaction solution was purified by preparative high performance liquid chromatography to obtain a preparation solution of the product. The preparation solution was freeze-dried to obtain 45 mg of the product; TOF-MS m / z (M+H)+ :1853.8. In a 25mL single-neck flask, compound 68 (40mg, 0.021mmol) was completely dissolved in 5mL of nitromethane, and then zinc bromide (97mg, 0.43mmol) was added, and the reaction was left at room temperature for 1 hour. The reaction process was monitored by HPLC. After the reaction was completed, the reaction solution was concentrated under reduced pressure by a water pump at 45℃ to obtain a crude product. The crude product was purified by preparative high performance liquid chromatography to obtain a preparation solution of the product. The preparation solution was freeze-dried and weighed to obtain 24mg of compound P-2; TOF-MS m / z(M+H) + :1698.7.

[0130] Example 66: Synthesis of compound P-3 [ka] 21 mg of compound P-3 was prepared according to the synthetic route of Example 6; TOF-MS m / z (M+H) + :1746.1.

[0131] Example 67: Synthesis of compound P-4 [ka] 26 mg of compound P-4 was prepared according to the synthetic route of Example 6; TOF-MS m / z (M+H) + :1966.3.

[0132] Example 68: Synthesis of Compound 69 [ka] Step 1: Synthesis of compound 69a In a 100 mL single-neck flask, compound 40 (10 g, 17.7 mmol) and compound VAPAB-OH (5.19 g, 17.7 mmol) were mixed and dissolved in 30 mL of DMF, then HATU (8.08 g, 21.2 mmol), HOBt (2.87 g, 21.2 mmol) and DIEA (4.38 mL, 26.5 mmol) were added, and the reaction was left at room temperature for 2 hours. After the reaction was found to be complete by HPLC monitoring, the reaction solution was purified by preparative liquid chromatography to obtain the preparation solution. The preparation solution was lyophilized to obtain compound 69a (9.3 g) in 62.6% yield, LCMS: [M+H] + =840.4. Step 2: Synthesis of compound 69b In a 100 mL single-neck flask, compound 69a (9 g, 10.7 mmol) and compound NPC (16.2 g, 53.5 mmol) were mixed and dissolved in 30 mL of DMF, then DIEA (3.24 mL, 32.1 mmol) was added, and the reaction was left at room temperature for 3 hours. After TLC monitoring showed that the reaction was complete, the reaction solution was purified by preparative liquid chromatography to obtain the preparation solution. The preparation solution was lyophilized to obtain compound 69b (8.23 g) in 76.5% yield, LCMS: [M+H] + =1005.4.

[0133] Step 3: Synthesis of compound 69c In a 25 mL single-neck flask, compound 69b (100 mg, 0.099 mmol), benzeneamine 2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]- and hydrochloride (1:1) (31.3 mg, 0.099 mmol) were mixed and dissolved in 5 mL of DMF, then DIEA (32.8 μL, 0.199 mmol) was added and the reaction was left at room temperature for 2 h. After HPLC monitoring showed that the reaction was complete, the reaction solution was purified by preparative high performance liquid chromatography to obtain the preparation solution. The preparation solution was lyophilized to obtain compound 69c (75.8 mg) in 64.5% yield, LCMS: [M+H] + =1181.5. Step 4: Synthesis of compound 69 In a 25 mL single-neck flask, compound 69c (75 mg, 0.063 mmol) was dissolved in 2 mL of DCM, then 4 mL of TFA was added and the reaction was left at room temperature for 2 h. After HPLC monitoring showed the reaction was complete, the solvent was removed under reduced pressure with a water pump at 45 °C to obtain a residue. The residue was purified by preparative high performance liquid chromatography to obtain the prepared solution. The prepared solution was lyophilized to obtain compound 69 (49.6 mg) in 76.3% yield, LCMS: [M+H] + =1025.4.

[0134] Example 69: Synthesis of compound P3-A [ka] Step 1: Synthesis of compound P3-A In a 25 mL one-neck flask, compound 69 (25 mg, 0.024 mmol) and cyclodextrin-N 3( 28.2 mg, 0.024 mmol) were mixed and dissolved in 5 mL of DMF and 2.5 mL of water, and then CuSO 4 5H 2 O (6.7 mg, 0.026 mmol) and sodium ascorbate (5.3 mg, 0.026 mmol) were added and the reaction was left at room temperature for 1 h. After the reaction was found to be complete by HPLC monitoring, the reaction solution was purified by preparative high performance liquid chromatography to obtain the prepared solution. The prepared solution was lyophilized to obtain compound P3-A (40.6 mg). TOF: [M+Na] + =2206.7.

[0135] Example 70: Synthesis of compound P3-B [ka] Step 1: Synthesis of compound P3-B In a 25 mL one-neck flask, compound 69 (25 mg, 0.024 mmol) and compound 18 (27.2 mg, 0.024 mmol) were mixed and dissolved in 5 mL of DMF and 2.5 mL of water, and then added with CuSO 4 5H 2O (6.7 mg, 0.026 mmol) and sodium ascorbate (5.3 mg, 0.026 mmol) were added and the reaction was left at room temperature for 1 h. After the reaction was found to be complete by HPLC monitoring, the reaction solution was purified by preparative high performance liquid chromatography to obtain the prepared solution. The prepared solution was lyophilized to obtain compound P3-B (34.5 mg). TOF: [M+H] + =2360.1.

[0136] Example 71: Synthesis of compound P3-C [ka] Step 1: Synthesis of compound 71 In a 100mL single-neck flask, compound 70 (5g, 9.7mmol) and compound NCP (14.7g, 48.5mmol) were mixed and dissolved in 30mL of DMF, then DIEA (4.9mL, 48.5mmol) was added, and the reaction was left at room temperature for 3 hours. The end point of the reaction was monitored by TLC. After the reaction was completed, the reaction solution was purified by preparative liquid chromatography to obtain a preparation solution. The preparation solution was freeze-dried to obtain compound 71 (7.66g), LCMS: [M+H] + =516.3. Step 2: Synthesis of compound 72 In a 25 mL single-neck flask, compound 71 (4 g, 7.76 mmol), benzeneamine, 2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]- and hydrochloride (1:1) (2.45 g, 7.76 mmol) were mixed and dissolved in 15 mL of DMF, after which DIEA (2.56 mL, 15.5 mmol) was added and the reaction was left at room temperature for 2 hours. After HPLC monitoring showed that the reaction was complete, the reaction solution was purified by preparative high performance liquid chromatography to obtain the prepared solution. The prepared solution was lyophilized to obtain compound 72 (4.1 g), LCMS: [M+H] + =857.5.

[0137] Step 3: Synthesis of compound 73 In a 50 mL single-neck flask, compound 72 (4 g) was dissolved in 27 mL of DMF, then diethylamine (3 mL) was added and the reaction was left at room temperature for 4 hours. After HPLC monitoring showed the reaction was complete, the reaction solution was concentrated under reduced pressure using an oil pump. The resulting yellow oil was purified by preparative high performance liquid chromatography to obtain the preparation solution. The preparation solution was lyophilized to obtain compound 73 (2.8 g), LCMS: [M+H] + =635.5. Step 4: Synthesis of compound P3-C In a 25 mL single-neck flask, compound 73 (80 mg, 0.126 mmol) was dissolved in 5 mL of DMF, then DIPEA (42 μL, 0.252 mmol) and compound 50 (0.126 mmol) were added, and the reaction was left at room temperature for 1 h. After HPLC monitoring showed the reaction to be complete, the reaction solution was concentrated under reduced pressure with a water pump at 45 °C to remove the solvent, and the resulting residue was sent directly to the next reaction. In a 25mL single-neck flask, the above crude compound was dissolved in 2mL of DCM, then 4mL of TFA was added, and the reaction was left at room temperature for 0.5 hours. After the reaction was found to be complete by HPLC monitoring, the reaction solution was concentrated under reduced pressure with a water pump at 45°C to remove the solvent and obtain a residue. The residue was purified by preparative high performance liquid chromatography to obtain the preparation solution. The preparation solution was lyophilized to obtain compound P3-C (37.7mg), TOF: [M+H] + =2146.8.

[0138] Example 72: Synthesis of compound P3-D [ka] Step 1: Synthesis of compound 75 Compound 73 (200 mg, 0.315 mmol) and 10 mL of DMF were added to a 25 mL one-neck flask, then compound 74 (134 mg, 0.315 mmol) was added at room temperature, HATU (240 mg, 0.62 mmol), HOBt (84 mg, 0.62 mmol) and DIPEA (0.12 mL) were added successively under cooling in an ice-water bath, and the resulting reaction solution was allowed to slowly warm to room temperature and react for 3 hours. After the reaction was found to be complete by TLC monitoring, the solvent was removed by rotary evaporation under reduced pressure. The concentrated residue was a pale yellow oily crude product. The crude product was sent directly to the next reaction without purification. The above compound was dissolved in 3 mL of dichloromethane, and the resulting solution was cooled to 0° C. under ice-water bath conditions, and TFA (1.5 mL) was slowly added dropwise. The resulting reaction solution was allowed to slowly warm to room temperature and react for 5 hours, and the reaction was monitored by HPLC. After the reaction was completed, the reaction solution was concentrated under reduced pressure at 45° C. to obtain a yellow oily crude product compound. The residue was purified by preparative high performance liquid chromatography to obtain the preparation solution. The preparation solution was freeze-dried to obtain compound 75 (162.0 mg), LC-MS: [M+H] + =986.6.

[0139] Step 2: Synthesis of compound 76 Compound 75 (80 mg, 0.081 mmol) and 10 mL of DMF were added to a 25 mL one-neck flask, then compound 23 (106 mg, 0.081 mmol) was added at room temperature, HATU (94 mg, 0.0.243 mmol), HOBt (16.5 mg, 0.122 mmol) and DIPEA (14 μL) were added sequentially under cooling in an ice-water bath, and the reaction was carried out for 2 hours. After the reaction was found to be complete by HPLC monitoring, the reaction solution was purified by preparative high performance liquid chromatography to obtain a preparation solution. The preparation solution was lyophilized to obtain an intermediate compound, which was directly sent to the next reaction. The intermediate was dissolved in 3 mL of DMF, and diethylamine (0.3 mL) was added dropwise at room temperature to react for 6 hours, and the reaction was monitored by HPLC. After the reaction was completed, the reaction solution was purified by preparative high performance liquid chromatography to obtain a preparation solution. The preparation solution was freeze-dried to obtain compound 76 (123.0 mg). TOF: [M+H] + =2055.1.

[0140] Step 3: Synthesis of compound P3-D Compound 76 (100 mg, 0.042 mmol) and 3 mL of DMF were added to a 10 mL PE tube, followed by compound 6 (17 mg, 0.042 mmol) at room temperature, followed by HATU (34 mg, 0.084 mmol), HOBt (12.5 mg, 0.084 mmol) and DIPEA (22 μL) in sequence under cooling in an ice-water bath, and the reaction was allowed to stand at room temperature for 1 h. After the reaction was found to be complete by HPLC monitoring, the solvent was removed by rotary evaporation under reduced pressure using an oil pump, and the residue was concentrated to obtain a pale yellow oily crude product. The crude product was sent directly to the next reaction without purification. The above crude product was dissolved in 2 mL of dichloromethane, the resulting solution was cooled to 0° C. under ice-water bath conditions, TFA (2 mL) was slowly added dropwise, and then the solution was allowed to slowly warm to room temperature and react for 2 hours. The reaction was monitored by HPLC. After the reaction was completed, the reaction solution was concentrated under reduced pressure at 45° C. to obtain a yellow oily crude product compound; the residue was purified by preparative high performance liquid chromatography to obtain the preparation solution. The preparation solution was freeze-dried to obtain compound P3-D (52.0 mg), TOF: [M+Na] + =2301.1.

[0141] Example 73: Synthesis of compound P3-E [ka] Compound P3-E (44 mg) was prepared according to the synthetic route of compound P3-D using compound 75 and compound 24 as starting materials. LC-MS: [M+H] + =1502.8.

[0142] Example 74: Synthesis of compound P3-F [ka] Compound P3-F (26 mg) was prepared according to the synthetic route of compound P3-D using compound 75 and compound 25 as starting materials. LC-MS: [M+H] + =1370.5.

[0143] Example 75: Synthesis of compound P3-G [ka] Compound P3-G (71 mg) was prepared according to the synthetic route of compound P3-A using compound 69 and compound 10 as starting materials. TOF: [M+H] + =1583.6.

[0144] Example 76: Synthesis of compound P3-H [ka] Step 1: Synthesis of compound PJA-PEG8-N3 In a 25 mL single-neck flask, meglumine (50 mg, 0.256 mmol), COOH-PEG-8-N3 (150 mg, 0.256 mmol) and 5 mL of DMF were added, followed by T3P (244 mg, 0.384 mmol, total 50%) at room temperature, and the reaction was left at room temperature for 5 hours. After the reaction was found to be complete by HPLC monitoring, the reaction solution was purified by preparative high performance liquid chromatography to obtain the preparation solution. The preparation solution was lyophilized to obtain compound PJA-PEG8-N3 (35 mg), LC-MS: [M+H] + =645.7. Step 2: Synthesis of compound P3-H In a 25 mL one-neck flask, compound 69 (25 mg, 0.024 mmol) and PJA-PEG 8 -N 3 (15.7 mg, 0.024 mmol) were mixed and dissolved in 5 mL of DMF and 2.5 mL of water, and then CuSO 4 5H2 O (6.7 mg, 0.026 mmol) and sodium ascorbate (5.3 mg, 0.026 mmol) were added and the reaction was left at room temperature for 1 h. After the reaction was found to be complete by HPLC monitoring, the reaction solution was purified by preparative high performance liquid chromatography to obtain the preparation solution. The preparation solution was lyophilized to obtain compound 2 (35 mg) in 78.6% yield, TOF: [M+H] + =1669.7.

[0145] Example 77: Synthesis of compound P3-I [ka] Compound P3-I (28 mg) was prepared according to the synthetic route of compound P3-A using compound 69 and compound 27 as starting materials. LC-MS: [M+H] + =1479.8.

[0146] Example 78: Synthesis of compound P3-J [ka] Step 1: Synthesis of compound 77 In a 25mL one-neck flask, maltose (2g, 5.85mmol) and Fmoc ethylenediamine (1.7g, 6mmol) were mixed and dissolved in 10mL of anhydrous tetrahydrofuran, then the resulting solution was cooled to 0°C under ice-water bath conditions, after which sodium borohydride (240mg, 6mmol) was added, and the resulting reaction solution was allowed to slowly warm to room temperature and react for 1 hour. After HPLC monitoring showed that the reaction was complete, 10mL of acetone was added while cooling in an ice-water bath to quench the reaction. Filtration through diatomaceous earth was performed. The filter cake was rinsed with 200mL of methanol, and the reaction solution was concentrated under reduced pressure and sent directly to the next reaction. In a 25mL single-neck flask, the above crude compound was dissolved in 8mL of DMF, then diethylamine (2mL) was added dropwise at room temperature, and the reaction was left at room temperature for 5 hours. After the reaction was found to be complete by HPLC monitoring, the reaction solution was purified by preparative high performance liquid chromatography to obtain the preparation solution. The preparation solution was lyophilized to obtain compound 77 (1.1g), LC-MS: [M+H] + =387.4. Step 2: Synthesis of compound P3-J Compound P3-J (45 mg) was prepared according to the synthetic route of compound P3-D using compound 75 and compound 77 as starting materials. LC-MS: [M+H] + =1356.9.

[0147] Example 79: Synthesis of compound P3-K [ka] Compound P3-K (39 mg) was prepared according to the synthetic route of compound P3-D using compound 75 and compound 28 as starting materials. LC-MS: [M+H] + =1488.4.

[0148] Example 80: Synthesis of compound P3-L [ka] Compound P3-L (22 mg) was prepared according to the synthetic route of compound P3-D using compound 75 and compound 30-PEG3 as starting materials. LC-MS: [M+H] + =1569.6.

[0149] Example 81: Synthesis of compound P3-M [ka] Compound P3-M (29 mg) was prepared according to the synthetic route of compound P3-D using compound 75 and compound 61 as starting materials. TOF: [M+H] + =1790.1.

[0150] Example 82: Synthesis of compound P3-N [ka] Compound P3-N (24 mg) was prepared according to the synthetic route of compound P3-D using compound 75 and compound 64 as starting materials. TOF: [M+H] + =1465.7.

[0151] Example 83: Synthesis of compound P3-O [ka] Compound P3-O (31 mg) was prepared according to the synthetic route of compound P3-D using compound 75 and compound 66 as starting materials. TOF: [M+H] + =1759.6.

[0152] Example 84: Synthesis of compound P3-P [ka] Compound P3-P (41 mg) was prepared according to the synthetic route of compound P3-A using compound 69 and compound 33 as starting materials. TOF: [M+H] + =1861.1.

[0153] Example 85: Synthesis of Compound P3-Q [ka] Compound P3-Q (36 mg) was prepared according to the synthetic route of compound P3-A using compound 69 and compound 34 as starting materials. TOF: [M+H] + =1861.1.

[0154] Example 86: Synthesis of compound P3-R [ka] Compound P3-R (18 mg) was prepared according to the synthetic route of compound P3-D using compound 75 and compound 35 as starting materials. TOF: [M+H] +=1455.9.

[0155] Example 87: Synthesis of compound P3-S [ka] Step 1: Synthesis of compound 78 In a 100 mL one-neck flask, mannuronic acid (5 g, 25.8 mmol) and propargylamine (11.1 g, 25.8 mmol) were mixed and completely dissolved in 50 mL of DMF, the resulting solution was cooled to 0 °C under ice-water bath conditions, and then EDCI (9.9 g, 51.6 mmol), HOBt (7 g, 51.6 mmol) and DIEA (12.7 mL, 77.4 mmol) were added, the resulting solution was allowed to naturally warm to room temperature and react for 2 hours. After the reaction was found to be complete by TLC monitoring, the reaction solution was poured into 100 mL of water and extracted three times with ethyl acetate (100 mL). * 3) The organic liquids were combined, washed twice with saturated NaCl solution, and then washed with anhydrous Na 2 SO 4 Drying at rt gave the crude compound which was carried on directly to the next reaction. The crude compound was added to a 250 mL single-neck flask and dissolved in 50 mL of DMF, followed by the addition of 10 mL of diethylamine, and the reaction was allowed to stand at room temperature for 2 hours. After TLC monitoring showed the reaction was complete, 500 mL of methyl tert-ether was added to the reaction solution, resulting in a white solid precipitate. The precipitate was then separated from the solution by filtration and dried to give compound 78 (3.96 g), LC-MS: [MH] + =369.5. Step 2: Synthesis of compound P3-S Compound P3-S (24 mg) was prepared according to the synthetic route of compound P3-D using compound 75 and compound 78 as starting materials. TOF: [M+H] + =1338.4.

[0156] Example 88: Synthesis of compound P3-T [ka] Step 1: Synthesis of compound 79 Following the synthetic route of compound 78, 13.2 g of compound 79 was prepared. LC-MS: [MH] + =237.5. Step 2: Synthesis of compound P3-T Compound P3-T (44 mg) was prepared according to the synthetic route of compound P3-D using compound 75 and compound 79 as starting materials. TOF: [M+H] + =1206.6.

[0157] Example 89: Synthesis of compound P3-U [ka] Step 1: Synthesis of Compound 80 In a 50 mL single-neck flask, mannuronic acid (1 g, 5.2 mmol) and propargylamine (2.2 g, 5.2 mmol) were mixed and completely dissolved in 10 mL of DMF, the resulting solution was cooled to 0°C under ice-water bath conditions, EDCI (2.0 g, 10.3 mmol), HOBt (1.4 g, 10.3 mmol) and DIEA (2.6 mL, 15.5 mmol) were added, and the reaction was left at room temperature for 2 hours. After the reaction was found to be complete by HPLC monitoring, the reaction solution was purified by preparative high performance liquid chromatography to obtain the prepared solution. The prepared solution was lyophilized to obtain compound 80 (0.97 g), LC-MS: [M+H] + =395.6. Step 2: Synthesis of compound P3-U Compound P3-U (16 mg) was prepared according to the synthetic route of compound P3-A using compound 69 and compound 80 as starting materials. TOF: [M+H] + =1419.8.

[0158] Example 90: Synthesis of compound P3-V [ka] Step 1: Synthesis of Compound 81 Compound 81 (0.49 g) was prepared according to the synthetic route of compound 80. LC-MS: [M+H] + =615.4. Step 2: Synthesis of compound P3-V Compound P3-V (30 mg) was prepared according to the synthetic route of compound P3-A using compound 69 and compound 81 as starting materials. TOF: [M+H] + =1639.9.

[0159] Example 91: Synthesis of compound P3-W [ka] Step 1: Synthesis of Compound 82 Compound 82 (2.17 g) was prepared according to the synthetic route of compound 80. LC-MS: [M+H] + =291.1. Step 2: Synthesis of compound P3-W Compound P3-W (35 mg) was prepared according to the synthetic route of compound P3-A using compound 69 and compound 82 as starting materials. TOF: [M+H] + =1315.8.

[0160] Example 92: Synthesis of Compounds P3-X2 [ka] Step 1: Synthesis of Compound 83 In a 25 mL single-neck flask, mono-6-O-(amino)-β-cyclodextrin (1 g, 0.88 mmol) and azido-ethylene glycol-acetic acid (180 mg, 0.88 mmol) were mixed and completely dissolved in 5 mL of DMF. The resulting solution was cooled to about 0° C. under ice-water bath conditions. Then, EDCI (386 mg, 2 mmol), HOBt (276 mg, 2 mmol) and DIEA (0.66 mL, 4 mmol) were added and the reaction was left at room temperature for 2 hours. After the reaction was found to be complete by HPLC monitoring, the reaction solution was purified by preparative high performance liquid chromatography to obtain the prepared solution. The prepared solution was lyophilized to obtain compound 83 (0.66 g), LC-MS: [M+H] + =1319.6. Step 2: Synthesis of Compound P3-X2 Compound P3-X2 (17 mg) was prepared according to the synthesis route of compound P3-A using compound 69 and compound 83 as starting materials. TOF: [M+Na] + =2365.9.

[0161] Example 93: Synthesis of Compound P3-Y8 [ka] Step 1: Synthesis of Compound 84 In a 25 mL one-neck flask, mono-6-O-(amino)-β-cyclodextrin (1 g, 0.88 mmol), 5,8,11,14,17,20,23,26-octaoxa-2-azanonaconicoic acid 1-(9H-fluoren-9-ylmethyl) ester (585.1 mg, 0.88 mmol) were completely dissolved in 5 mL of DMF. The resulting solution was cooled to about 0 °C in ice-water bath conditions, EDCI (388 mg, 2 mmol), HOBt (275 mg, 2 mmol) and DIEA (0.66 mL, 4 mmol) were added, and the reaction was left at room temperature for 2 hours. After the reaction was found to be complete by HPLC monitoring, the reaction solution was concentrated under reduced pressure by oil pump and sent directly to the next reaction. In a 25mL single-neck flask, the above crude compound was added and dissolved in 10mL of DMF, then 2mL of diethylamine was added, and the reaction was left at room temperature for 2 hours. After the reaction was found to be complete by HPLC monitoring, the reaction solution was purified by preparative high performance liquid chromatography to obtain the preparation solution. The preparation solution was lyophilized to obtain compound 84 (0.87g), TOF: [M+H] + =1557.5. Step 2: Synthesis of compound P3-Y8 Compound P3-Y8 (21 mg) was prepared according to the synthesis route of compound P3-D using compound 75 and compound 84 as starting materials. TOF: [M+Na] + =2549.1.

[0162] Example 94: Synthesis of Compounds P3-Z4 [ka] Step 1: Synthesis of Compound 85 Compound 85 (0.91 g) was prepared according to the synthetic route of compound 84. LC-MS: [M+H] + =1233.3. Step 2: Synthesis of compound P3-Y8 Compound P3-Z4 (33 mg) was prepared according to the synthetic route of compound P3-D using compound 75 and compound 85 as starting materials. TOF: [M+H] + =2203.1.

[0163] Example 95: Synthesis of Compound 86 [ka] Step 1: Synthesis of compound 86b To obtain compound 86a, please refer to the synthesis of "compound 26" in patent application CN102933236A. In a 250 mL three-neck flask, add N 2 Compound 86a (3 g, 6.7 mmol) and 50 mL of anhydrous THF were added in the presence of , the temperature was maintained at 0 °C with an ice-water bath, and solid sodium borohydride (384 mg, 10.2 mmol) was added. The reaction solution was reacted at 0 °C for 30 min, then allowed to naturally warm to room temperature and react for 2 h. After TLC monitoring showed the reaction was complete, the reaction solution was cooled with ice water, 50 mL of water was slowly added to quench the reaction, and 1N dilute HCl was added until no bubbles were observed. The resulting solution was dissolved in ethyl acetate (60 mL). * 3) Extract with 100 ml of NaCl; combine the organic layers, wash twice with saturated NaCl solution, and 2 SO 4 The crude product was purified by column chromatography (DCM / MeOH=20:1 to 5:1) to give compound 86b (2.08 g, yellow solid) in 74.3% yield. LC-MS: [M+H] + =415.1. Step 2: Synthesis of compound 86c In a 100 mL single-neck flask, compound 86b (2 g, 4.8 mmol), TBSCl (1.32 g, 8.8 mmol), and imidazole (1.2 g, 17.6 mmol) were mixed and dissolved in 15 mL of anhydrous DMF, and the reaction was allowed to stand at room temperature for 3 h. After TLC monitoring showed the reaction was complete, the reaction solution was poured into 30 mL of water and diluted with DCM (25 mL). * 3) Extract with 100 mL of sodium chloride; combine the organic layers, wash with water and brine, and 2 SO 4 The crude product was subjected to column chromatography (PE / EA=5:1 to 1:2) to give compound 86c (2.28 g, yellow solid) in 89.3% yield. LC-MS: [M+H] + =529.2.

[0164] Step 3: Synthesis of compound 86d In a 100 mL single-neck flask, compound 3f (2.2 g, 4.16 mmol) and 30 mL of 5% formic acid / methanol solution were mixed and cooled to less than 5° C. in an ice bath, zinc powder (5.44 g, 83.2 mmol) was added slowly with stirring, and the reaction was allowed to stand at room temperature for 1 h. After TLC monitoring showed the reaction was complete, the reaction solution was filtered while the reaction solution was hot. The filter cake was washed with a small amount of methanol. The filtrated was adjusted to pH 7 with saturated sodium bicarbonate solution, then concentrated under reduced pressure at 45° C. to remove the solvent, giving a brown oil. 60 mL of DCM was added, the organic layer was separated, washed once with saturated salt solution, and diluted with anhydrous Na 2 SO 4 The crude product was purified by column chromatography (PE:EA=1:1-1:3) to give compound 86d (1.87 g, pale yellow solid) in 90.1% yield. LC-MS: [M+H] + =500.2. Step 4: Synthesis of compound 86e In a 50 mL single-neck flask, compound 3g (1.8 g, 3.6 mmol) and Fmoc-VA-PABO-PNP (2.45 g, 3.6 mmol) were mixed and completely dissolved in 15 mL of DMF, then DIEA (893 μL, 5.4 mmol) was added and the reaction was left at room temperature for 2 hours. After the reaction was found to be complete by HPLC monitoring, the reaction solution was purified by preparative high performance liquid chromatography to obtain the preparation solution. The preparation solution was lyophilized to obtain compound 86e (2.94 g, pale yellow solid) in 78.3% yield, LC-MS: [M+H] + =1040.5.

[0165] Step 5: Synthesis of compound 86f In a 50 mL single-neck flask, compound 86e (2.9 g, 2.78 mmol) was dissolved in 8 mL of THF and 8 mL of water, then 20 mL of glacial acetic acid was added and the reaction was allowed to stand at room temperature. After the reaction was found to be complete by HPLC monitoring, the reaction solution was slowly dripped into 400 mL of saturated sodium bicarbonate solution and ethyl acetate (100 mL * 3), extract with 100 ml of water, combine the organic liquids, wash with water and brine, and add anhydrous Na 2 SO 4 The crude product was subjected to column chromatography (DCM / MeOH=10:1 to 5:1) to give compound 86f (1.94 g, pale yellow solid) in 75% yield. LC-MS: [M+H] + =926.3. Step 6: Synthesis of compound 86g In a 100 mL three-neck flask, compound 86f (1.9 g, 2.0 mmol) was dissolved in 40 mL of anhydrous DCM and heated with N 2 Dess-Martin high iodine reagent (0.93 g, 2.2 mmol) was added in the presence of 1,000,000 sulphate, and the reaction was allowed to stand at room temperature for 4 hours. After the reaction was found to be complete as determined by TLC monitoring, the reaction solution was filtered, and the filtrate was washed with saturated sodium bicarbonate solution, water and saturated NaCl solution, and then washed with anhydrous Na 2 SO 4The crude product was purified by preparative high performance liquid chromatography to obtain a crude solution of the product. The crude solution was freeze-dried to obtain compound 86g (1.61 g, a kind of white solid) in 85% yield. LC-MS: [M+H] + =924.4.

[0166] Step 7: Synthesis of compound 86h In a 50 mL single-neck flask, compound 86g (1.5 g, 1.62 mmol) was completely dissolved in 15 mL of DMF, then 3 mL of diethylamine was added, and the reaction was left at room temperature for 1 h. After TLC monitoring showed that the reaction was complete, the reaction solution was poured into 100 mL of methyl tert-ether to obtain a solid precipitate. The precipitate was separated from the solution by filtration to obtain the deprotected product. The deprotected product was dissolved in 15 mL of DMF, compound 40 (0.92 g, 1.62 mmol) and EEDQ (0.48 g, 1.94 mmol) were added, and the reaction was left at room temperature for 2 h. After HPLC monitoring showed that the reaction was complete, the reaction solution was purified by preparative high performance liquid chromatography to obtain the prepared solution. The prepared solution was lyophilized to obtain compound 86h (1.30 g, a kind of white solid) in 64.2% yield, LC-MS: [M+H] + =1248.5. Step 8: Synthesis of Compound 86 In a 50 mL single-neck flask, compound 86h (500 mg, 0.4 mmol) was dissolved in 5 mL of DCM, then trifluoroacetic acid (5 mL) was added and the reaction was left at room temperature for 1 h. After HPLC monitoring showed the reaction was complete, the reaction solution was concentrated under reduced pressure at 45° C. to remove the solvent and obtain a residue. The residue was purified by preparative high performance liquid chromatography to obtain the preparation solution. The preparation solution was lyophilized to obtain compound 86 (294 mg) in 67.4% yield, LC-MS: [M+H] + =1092.4.

[0167] Example 96: Synthesis of compound P4-A [ka] Step 1: Synthesis of compound P4-A In a 25 mL single-neck flask, compound 86 (25 mg, 0.023 mmol) and cyclodextrin-N 3( 26.5 mg, 0.023 mmol) were mixed and dissolved in 5 mL of DMF and 2.5 mL of water, and then CuSO 4 5H 2 O (6.7 mg, 0.026 mmol) and sodium ascorbate (5.3 mg, 0.026 mmol) were added. The reaction was left at room temperature for 1 h. After the reaction was found to be complete by HPLC monitoring, the reaction solution was purified by preparative high performance liquid chromatography to obtain the prepared solution. The prepared solution was lyophilized to obtain compound P4-A (36.9 mg) in 71.6% yield, TOF: [M+H] + =2251.8.

[0168] Example 97: Synthesis of compound P4-B [ka] Step 1: Synthesis of compound P4-B In a 25 mL one-neck flask, compound 86 (25 mg, 0.023 mmol) and compound 18 (30.7 mg, 0.023 mmol) were mixed and dissolved in 5 mL of DMF and 2.5 mL of water, and then added with CuSO 4 5H 2 O (6.6 mg, 0.026 mmol) and sodium ascorbate (5.3 mg, 0.026 mmol) were added. The reaction was left at room temperature for 1 h. After the reaction was found to be complete by HPLC monitoring, the reaction solution was purified by preparative high performance liquid chromatography to obtain the prepared solution. The prepared solution was lyophilized to obtain compound P4-B (37.2 mg) in 66.6% yield, TOF: [M+H] + =2427.2.

[0169] Example 98: Synthesis of compound P4-C [ka] Step 1: Synthesis of Compound 88 In a 50 mL single-neck flask, compound 86g (2 g, 2.16 mmol) was dissolved in 16 mL of DMF, then diethylamine (4 mL) was added and the reaction was left at room temperature for 8 hours. After HPLC monitoring showed the reaction to be complete, the reaction solution was concentrated under reduced pressure using an oil pump, and the resulting yellow oil was concentrated using an oil pump to remove the solvent to give compound 88, which was sent directly to the next reaction. The above crude compound was dissolved in 10 mL of DMF, then DIPEA (714 μL, 4.32 mmol) was added at room temperature, followed by compound 50 (4 g, 2.16 mmol), and the reaction was left at room temperature for 3 hours. After the reaction was found to be complete by HPLC monitoring, the reaction solution was purified by preparative high performance liquid chromatography to obtain the preparation solution. The preparation solution was lyophilized to obtain compound 88 (3.9 g), TOF: [M+H] + =2370.3. Step 2: Synthesis of compound P4-C In a 25mL single-neck flask, compound 88 (1g, 0.42mmol) and 4mL DCM were mixed and completely dissolved, then 4mL TFA was added and the reaction was left at room temperature for 0.5h. After the reaction was found to be complete by HPLC monitoring, the reaction solution was concentrated by water pump at 45°C to remove the solvent and obtain a residue. The residue was purified by preparative high performance liquid chromatography to obtain the preparation solution. The preparation solution was freeze-dried to obtain compound P4-C (577mg), TOF:[M+H] + =2214.0.

[0170] Example 99: Synthesis of compound P4-D [ka] Step 1: Synthesis of Compound 89 In a 25mL one-neck flask, compound 87 (300mg, 0.43mmol) and 10mL of DMF were added, followed by compound 74 (183mg, 0.43mmol) at room temperature. HATU (333mg, 0.86mmol), HOBt (118mg, 0.86mmol) and DIPEA (0.22mL) were added successively under cooling with ice-water bath, and the resulting solution was allowed to slowly warm to room temperature and react for 5 hours. After TLC monitoring showed the reaction was complete, the solvent was removed by rotary evaporation under reduced pressure. The residue was concentrated to give a pale yellow oily crude product. The crude product was directly sent to the next reaction without purification. The above compound was dissolved in 3 mL of dichloromethane, and the resulting solution was cooled to 0° C. under ice-water bath conditions, and TFA (3 mL) was slowly added dropwise, and the resulting reaction solution was allowed to slowly warm to room temperature and react for 0.5 hours. After the reaction was found to be complete by HPLC monitoring, the reaction solution was concentrated under reduced pressure at 45° C. to obtain a yellow oily crude compound. The residue was purified by preparative high performance liquid chromatography to obtain the preparation solution. The preparation solution was lyophilized to obtain compound 89 (266 mg), LC-MS: [M+H] + =1053.5. Step 2: Synthesis of Compound 90 Compound 89 (200 mg, 0.19 mmol) and 10 mL of DMF were added to a 25 mL one-neck flask, followed by compound 23 (484 mg, 0.19 mmol) at room temperature. HATU (156 mg, 0.4 mmol), HOBt (82 mg, 0.6 mmol) and DIPEA (100 μL) were added sequentially under cooling in an ice-water bath and reacted for 3 hours. After the reaction was found to be complete by HPLC monitoring, the reaction solution was purified by preparative high performance liquid chromatography to obtain a preparation solution. The preparation solution was lyophilized, and the intermediate compound was sent directly to the next reaction. The above intermediate was dissolved in 3 mL of DMF, then diethylamine (0.6 mL) was added dropwise at room temperature, and the reaction was left at room temperature for 5 hours. The reaction was monitored by HPLC. After the reaction was completed, the reaction solution was purified by preparative high performance liquid chromatography to obtain the preparation solution. The preparation solution was freeze-dried to obtain compound 90 (301 mg), TOF: [M+H] + =2122.1.

[0171] Step 3: Synthesis of Compound 91 In a 25 mL single-neck flask, compound 90 (200 mg, 0.094 mmol) and 5 mL of DMF were added, followed by compound 6 (38 mg, 0.042 mmol) at room temperature. HATU (76 mg, 0.188 mmol), HOBt (29 mg, 0.188 mmol) and DIPEA (64 μL) were added sequentially under cooling in an ice-water bath, and the reaction was allowed to stand at room temperature for 2 hours. After the reaction was found to be complete by HPLC monitoring, the reaction solution was purified by preparative high performance liquid chromatography to obtain the preparation solution. The preparation solution was lyophilized to obtain compound 91 (198 mg), TOF: [M+H] + =2502.2. Step 4: Synthesis of compound P4-D Compound 91 (100 mg, 0.04 mmol) was dissolved in 2 mL of dichloromethane, and the resulting solution was cooled to 0° C. under ice-water bath conditions. TFA (2 mL) was slowly added dropwise, and the resulting solution was allowed to slowly warm to room temperature and react for 0.5 h. The reaction was monitored by HPLC. After the reaction was completed, the reaction solution was concentrated under reduced pressure at 45° C. to obtain a yellow oily crude compound. The residue was purified by preparative high performance liquid chromatography to obtain the preparation solution. The preparation solution was freeze-dried to obtain compound P4-D (62.0 mg), TOF: [M+Na] + =2368.0.

[0172] Example 100: Synthesis of compound P4-E [ka] Compound P4-E (37 mg) was prepared according to the synthetic route of compound P4-D using compound 89 and compound 24 as starting materials. LC-MS: [M+H] + =1569.6.

[0173] Example 101: Synthesis of compound P4-F [ka] Compound P4-F (41 mg) was prepared according to the synthetic route of compound P4-D using compound 89 and compound 25 as starting materials. LC-MS: [M+H] + =1437.5.

[0174] Example 102: Synthesis of compound P4-G [ka] Compound P4-G (25 mg) was prepared according to the synthetic route of compound P4-A using compound 86 and compound 10 as starting materials. LC-MS: [M+H] + =1650.6.

[0175] Example 103: Synthesis of compound P4-H [ka] Compound P4-H (34 mg) was prepared according to the synthetic route of compound P4-A using compound 86 and compound PJA-PEG8-N3 as starting materials. LC-MS: [M+H] + =1736.8.

[0176] Example 104: Synthesis of compound P4-I [ka] Compound P4-I (55 mg) was prepared according to the synthetic route of compound P4-A using compound 86 and compound 27 as starting materials. LC-MS: [M+H] + =1546.9.

[0177] Example 105: Synthesis of Compound P4-J [ka] Compound P4-J (31 mg) was prepared according to the synthetic route of compound P4-D using compound 89 and compound 77 as starting materials. LC-MS: [M+H] + =1423.7.

[0178] Example 106: Synthesis of compound P4-K [ka] Starting from compound 89 and compound 28, compound P4-K (24 mg) was prepared according to the synthetic route of compound P4-D, LC-MS: [M+H]+=1555.7.

[0179] Example 107: Synthesis of compound P4-L [ka] Compound P4-L (24 mg) was prepared according to the synthetic route of compound P4-D using compound 89 and compound 30 as starting materials. LC-MS: [M+H] + =1636.6.

[0180] Example 108: Synthesis of compound P4-M [ka] Compound P4-M (20 mg) was prepared according to the synthetic route of compound P4-D using compound 89 and compound 61 as starting materials. LC-MS: [M+H] + =1857.0.

[0181] Example 109: Synthesis of compound P4-N [ka] Compound P4-N (17 mg) was prepared according to the synthetic route of compound P4-D using compound 89 and compound 64 as starting materials. LC-MS: [M+H] + =1532.7.

[0182] Example 110: Synthesis of compound P4-O [ka] Compound P4-O (32 mg) was prepared according to the synthetic route of compound P4-D using compound 89 and compound 66 as starting materials. LC-MS: [M+H] + =1626.8.

[0183] Example 111: Synthesis of compound P4-P [ka] Compound P4-P (42 mg) was prepared according to the synthetic route of compound P4-A using compound 86 and compound 33 as starting materials. LC-MS: [M+H] + =1928.0.

[0184] Example 112: Synthesis of Compound P4-Q [ka] Compound P4-Q (35 mg) was prepared according to the synthetic route of compound P4-A using compound 86 and compound 34 as starting materials. LC-MS: [M+H] + =1603.6.

[0185] Example 113: Synthesis of compound P4-R [ka] Compound P4-R (31 mg) was prepared according to the synthetic route of compound P4-D using compound 89 and compound 35 as starting materials. LC-MS: [M+H] + =1522.8.

[0186] Example 114: Synthesis of compound P4-S [ka] Compound P4-S (21 mg) was prepared according to the synthetic route of compound P4-D using compound 89 and compound 78 as starting materials. LC-MS: [M+H] + =1405.5.

[0187] Example 115: Synthesis of compound P4-T [ka] Compound P4-T (31 mg) was prepared according to the synthetic route of compound P4-D using compound 89 and compound 79 as starting materials. LC-MS: [M+H] + =1273.4.

[0188] Example 116: Synthesis of compound P4-U [ka] Compound P4-U (47 mg) was prepared according to the synthetic route of compound P4-A using compound 86 and compound 80 as starting materials. LC-MS: [M+H] + =1486.7.

[0189] Example 117: Synthesis of compound P4-V [ka] Compound P4-V (40 mg) was prepared according to the synthetic route of compound P4-A using compound 86 and compound 81 as starting materials. LC-MS: [M+H] + =1706.7.

[0190] Example 118: Synthesis of compound P4-W [ka] Compound P4-W (37 mg) was prepared according to the synthetic route of compound P4-A using compound 86 and compound 82 as starting materials. LC-MS: [M+H] + =1382.9.

[0191] Example 119: Synthesis of Compound P4-X2 [ka] Compound P4-X2 (22 mg) was prepared according to the synthetic route of compound P4-A using compound 86 and compound 83 as starting materials. TOF: [M+H] + =2411.2.

[0192] Example 120: Synthesis of compound P4-Y8 [ka] Compound P4-Y8 (29 mg) was prepared according to the synthetic route of compound P4-D using compound 89 and compound 84 as starting materials. TOF: [M+H] + =2594.5.

[0193] Example 121: Synthesis of Compound P4-Z4 [ka] Compound P4-Z4 (28 mg) was prepared according to the synthetic route of compound P4-D using compound 89 and compound 85 as starting materials. TOF: [M+Na] + =2292.1. The antibodies were prepared according to conventional antibody preparation methods, such as vector construction followed by purification and expression by transfection into eukaryotic cells. The sequence of the anti-Trop-2 antibody is shown below: Light chain (SEQ ID NO: 1) DIQMTQSPSSLSASVGDRVTITCRASQDINKYLAWYQQKPGKVPKLLIYSTSTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCLQYDDLFTFGQGTKLEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Heavy Chain (SEQ ID NO:2) QVQLVQSGAEVKKPGASVKLSCKASGYTFTSFDINWVRQAPEQRLEWMGWIFPGDGNTKYSQKFQGRATITRDTSASTAYMELSSLRSEDTAVYYCVRGEALYYFDYWGQG TLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG The sequence of the anti-CD33 antibody is shown below: Light chain (SEQ ID NO:3) DIQLTQSPSTLSASVGDRVTITCRASESLDNYGIRFLTWFQQKPGKAPKLLMYAASNQGSGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQTKEVPWSFGQGTKVE VKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVCTKSFNRGEC Heavy Chain (SEQ ID NO:4) EVQLVQSGAEVKKPGSSVKVSCKASGYTITDSNIHWVRQAPGQSLEWIGYIYPYNGGTDYNQKFKNRATLTVDNPTNTAYMELSSLRSEDTAFYYCVNGNPWLAYWGQGTL VTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKT HTCPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG In the following examples, anti-CD33 antibodies were used to prepare ligand-drug conjugates. Ligand-drug conjugates containing anti-TROP-2 antibodies were prepared according to the preparation methods disclosed in these examples.

[0194] Example 122: Preparation of Ligand Drug Conjugates: General join method After preliminary purification, antibody molecules L with a monomer ratio of more than 95% were transferred to phosphate buffer solution at a concentration of 10 mg / mL using ultrafiltration centrifuge tubes. TCEP was added in an amount 20 times the antibody mole number and reacted at room temperature for 4 hours to open the interchain disulfide bond of the antibody. Linker drug compound (payload) was added in an amount 20 times the antibody mole number and reacted at room temperature for 2 hours. After the reaction was completed, the reaction solution was transferred to PBS using a filtration centrifuge tube with a molecular weight cutoff of 30 KDa to remove unbound payload. The ADC sample after solution exchange was filtered using a 0.22 μm sterilization filter for later use.

[0195] Example 123: [ka] ADC-1 was synthesized according to the general method for the preparation of ligand-drug conjugates.

[0196] Example 124: [ka] ADC-2 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0197] Example 125: [ka] ADC-3 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0198] Example 126: [ka] ADC-4 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0199] Example 127: [ka] ADC-5 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0200] Example 128: [ka] ADC-6 was synthesized according to the general method for the preparation of ligand-drug conjugates.

[0201] Example 129: [ka] ADC-7 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0202] Example 130: [ka] ADC-8 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0203] Example 131: [ka] ADC-9 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0204] Example 132: [ka] ADC-10 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0205] Example 133: [ka] ADC-11 was synthesized according to the general method for the preparation of ligand-drug conjugates.

[0206] Example 134: [ka] ADC-12 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0207] Example 135: [ka] ADC-13 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0208] Example 136: [ka] ADC-14 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0209] Example 137: [ka] ADC-15 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0210] Example 138: [ka] ADC-16 was synthesized according to the general method for the preparation of ligand-drug conjugates.

[0211] Example 139: [ka] ADC-17 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0212] Example 140: [ka] ADC-18 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0213] Example 141: [ka] ADC-19 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0214] Example 142: [ka] ADC-20 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0215] Example 143: [ka] ADC-21 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0216] Example 144: [ka] ADC-22 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0217] Example 145: [ka] ADC-23 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0218] Example 146: [ka] ADC-24 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0219] Example 147: [ka] ADC-25 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0220] Example 148: [ka] ADC-26 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0221] Example 149: [ka] ADC-27 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0222] Example 150: [ka] ADC-28 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0223] Example 151: [ka] ADC-29 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0224] Example 152: [ka] ADC-30 was synthesized according to the general method for preparation of ligand drug conjugates.

[0225] Example 153: [ka] ADC-31 was synthesized according to the general method for the preparation of ligand-drug conjugates.

[0226] Example 154: [ka] ADC-32 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0227] Example 155: [ka] ADC-33 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0228] Example 156: [ka] ADC-34 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0229] Example 157: [ka] ADC-35 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0230] Example 158: [ka] ADC-36 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0231] Example 159: [ka] ADC-37 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0232] Example 160: [ka] ADC-38 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0233] Example 161: [ka] ADC-39 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0234] Example 162: [ka] ADC-40 was synthesized according to the general method for preparation of ligand drug conjugates.

[0235] Example 163: [ka] ADC-41 was synthesized according to the general method for preparation of ligand drug conjugates.

[0236] Example 164: [ka] ADC-42 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0237] Example 165: [ka] ADC-43 was synthesized according to the general method for preparation of ligand drug conjugates.

[0238] Example 166: [ka] ADC-44 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0239] Example 167: [ka] ADC-45 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0240] Example 168: [ka] ADC-46 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0241] Example 169: [ka] ADC-47 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0242] Example 170: [ka] ADC-48 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0243] Example 171: [ka] ADC-49 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0244] Example 172: [ka] ADC-50 was synthesized according to the general method for preparation of ligand drug conjugates.

[0245] Example 173: [ka] ADC-51 was synthesized according to the general method for preparation of ligand drug conjugates.

[0246] Example 174: [ka] ADC-52 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0247] Example 175: [ka] ADC-53 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0248] Example 176: [ka] ADC-54 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0249] Example 177: [ka] ADC-55 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0250] Example 178: [ka] ADC-56 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0251] Example 179: [ka] ADC-57 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0252] Example 180: [ka] ADC-58 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0253] Example 181: [ka] ADC-59 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0254] Example 182: [ka] ADC-60 was synthesized according to the general method for preparation of ligand drug conjugates.

[0255] Example 183: [ka] ADC-61 was synthesized according to the general method for the preparation of ligand-drug conjugates.

[0256] Example 184: [ka] ADC-62 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0257] Example 185: [ka] ADC-63 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0258] Example 186: [ka] ADC-64 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0259] Example 187: [ka] ADC-65 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0260] Example 188: [ka] ADC-66 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0261] Example 189: [ka] ADC-68 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0262] Example 190: [ka] ADC-68 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0263] Example 191: [ka] ADC-69 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0264] Example 192: [ka] ADC-70 was synthesized according to the general method for preparation of ligand drug conjugates.

[0265] Example 193: [ka] ADC-71 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0266] Example 194: [ka] ADC-72 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0267] Example 195: [ka] ADC-73 was synthesized according to the general method for preparation of ligand drug conjugates.

[0268] Example 196: [ka] ADC-74 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0269] Example 197: [ka] ADC-75 was synthesized according to the general method for preparation of ligand drug conjugates.

[0270] Example 198: [ka] ADC-76 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0271] Example 199: [ka] ADC-77 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0272] Example 200: [ka] ADC-78 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0273] Example 201: [ka] ADC-79 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0274] Example 202: [ka] ADC-80 was synthesized according to the general method for preparation of ligand drug conjugates.

[0275] Example 203: [ka] ADC-81 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0276] Example 204: [ka] ADC-82 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0277] Example 205: [ka] ADC-83 was synthesized according to the general method for preparation of ligand drug conjugates.

[0278] Example 206: [ka] ADC-84 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0279] Example 207: [ka] ADC-85 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0280] Example 208: [ka] ADC-86 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0281] Example 209: [ka] ADC-87 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0282] Example 210: [ka] ADC-88 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0283] Example 211: [ka] ADC-89 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0284] Example 212: [ka] ADC-90 was synthesized according to the general method for preparation of ligand drug conjugates.

[0285] Example 213: [ka] ADC-91 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0286] Example 214: [ka] ADC-92 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0287] Example 215: [ka] ADC-93 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0288] Example 216: [ka] ADC-94 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0289] Example 217: [ka] ADC-95 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0290] Example 218: [ka] ADC-96 was synthesized according to the general method for preparation of ligand drug conjugates.

[0291] Example 219: [ka] ADC-97 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0292] Example 220: [ka] ADC-98 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0293] Example 221: [ka] ADC-99 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0294] Example 222: [ka] ADC-100 was synthesized according to the general method for preparation of ligand drug conjugates.

[0295] Example 223: [ka] ADC-101 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0296] Example 224: [ka] ADC-102 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0297] Example 225: [ka] ADC-103 was synthesized according to the general method for preparation of ligand drug conjugates.

[0298] Example 226: [ka] ADC-104 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0299] Example 227: [ka] ADC-105 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0300] Example 228: [ka] ADC-106 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0301] Example 229: [ka] ADC-107 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0302] Example 230: [ka] ADC-108 was synthesized according to the general method for the preparation of ligand drug conjugates.

[0303] Example 231: Control ADC-109 [ka] The ADC-109 payload was prepared according to the synthetic route of Example 11 of patent application "WO2018233571A1", and the ligand-drug conjugate was prepared according to the general method of the present invention.

[0304] Example 232: Control ADC-110 [ka] The payload of ADC-110 was prepared according to the synthetic route of Example 4-3 on page 148 of the patent application "WO2019195665A1", and the ligand-drug conjugate was prepared according to the general method of the present invention.

[0305] Example 233: Control ADC-111 [ka] The payload of ADC-111 was prepared according to the synthetic route of the compound of formula (14) in patent application WO2019044946A1, and the ligand-drug conjugate was prepared according to the general method of the present invention.

[0306] Example 234: Determination of Monomer Ratio of ADC 1) Determination of monomer ratio using SEC-HPLC: Chromatography column: Biocore SEC-300 5 μm, 4.6 × 300 mm; Manufacturer: NanoChrom, Product code: B213-050030-04630S; Mobile phase: 50 mM PB + 300 mM NaCl + 200 mM Arg + 5% IPA, pH = 6.5 [Table 1] 2) Results: [Table 2] 1A-1C are SEC-HPLC spectra of ADC-29, ADC-56, and ADC-111, respectively, showing aggregation of the ADCs. 3) Conclusion: The ADC of the present invention has excellent properties, being characterized by low degradation and low aggregation, and having a high monomer ratio.

[0307] Example 235: Determination of DAR 1) Method: RP-HPLC was used to determine DAR: Chromatography column: Proteomix RP-1000 4.6 x 100 mm 5 μm 1000A; Manufacturer: Sepax Product code: 465950-4610 [Table 3] 2) Results: [Table 4] 2A-2C are RP-HPLC spectra of ADC-29, ADC-56, and ADC-111, respectively, showing the DAR of the ADCs. 3) Conclusion: The ADC of the present invention has the excellent property of having a high DAR, and significantly increased the drug concentration at the target site at the same dose of ADC.

[0308] Example 236: Plasma Stability 1) Operation A fixed amount of ADC samples was spiked into human IgG-depleted human plasma. There were three tubes for each ADC. Incubation was performed in a 37°C water bath. After 7, 14, and 21 days of incubation, ADC samples were taken and 100uL of Protein A resin (MabSelect SuReTM LX Lot:#10221479GE, washed with PBS) was added to each tube. After adsorption by shaking on a vertical mixer for 2 hours, washing and elution steps were performed to obtain incubated ADC samples. The ADC samples incubated for the indicated times were tested by RP-HPLC. 2) Results: [Table 5] 3) Conclusion Compared to the control ADC-109, the hydrophilicity-enhanced ADCs of the invention, such as ADC-1 and ADC-28, had higher monomer ratios and less aggregation in plasma after incubation in plasma at 37° C. for 21 days, indicating that the ADCs of the invention have better hydrophilicity and stability.

[0309] Example 237: In vitro efficacy testing of ADCs 1) Method: In this example, various human tumor cell lines (A431 (human epidermoid carcinoma cells), HL-60 (human promyelocytic acute leukemia cells), TF-1 (human hematologic leukemia cells), HEL92.1.7 (human erythroleukemia cells), MV4-11 (human myelomonocytic leukemia cells), and MOLM-13 (human acute myeloid leukemia cells)) were used as test models to evaluate the in vitro efficacy of ADCs. A certain number of tumor cell lines were seeded in a 96-well plate, and gradient dilutions of the test antibodies and the corresponding ADCs were added to the cells, and after 5 days, cell viability was examined using Alamar Blue or MTS. The inhibitory effects of the test antibodies and ADCs on the tumor cell lines were evaluated using IC 50 The antibody drug was diluted 7-fold with a starting concentration of 500 nM for a total of 8 concentration points, and treatment was performed for 5 days. The final algorithm was based on Viability = (Experimental group - Blank) / (Control - Blank) x 100%, followed by curve fitting using Graph Pad Prism to calculate the inhibitory concentration (IC 50 ) Median was calculated. 2) Results: [Table 6] 3) Conclusion: The results are shown in Table 6 and Figures 3A to 3F. In in vitro efficacy studies in various human tumor cell lines (A431, HL-60, TF-1, HEL92.1.7, MV4-11 and MOLM-13), the ADCs of the invention show excellent activity.

[0310] Example 238: In vivo efficacy testing of ADCs The present invention established a subcutaneously transplanted tumor model in HEL92.1.7 BALB / c nude mice to evaluate the in vivo efficacy of ADCs. 1) Method: 5×10 6HEL92.1.7 cells (0.1 mL / mouse) were subcutaneously injected into the right scapula of 5-6 week-old BALB / c nude mice. The average tumor size in mice was approximately 230-270 mm. 3 At age 18, mice were randomly assigned to vehicle control group (vehicle), monoclonal antibody (CD33-mAb) treatment group (10 mg / kg), and ADC treatment group (5 mg / kg, 10 mg / kg), with 5 mice per group, and administered the drug (D0) via tail vein injection at 10 mL / kg body weight. 2) Results: [Table 7] [Table 8] 3) Conclusion: As shown in Table 7, compared to the blank control and monoclonal antibody-treated groups, the ADCs of the present invention significantly inhibited tumor growth at various doses, showing dose-dependence to some extent. As shown in Table 8, the body weight of the test animals in each group was normal without significant changes during administration at various doses, indicating that the ADC of the present invention has no obvious toxic or side effects.

[0311] Although specific embodiments of the present invention have been described in detail, it will be understood that those skilled in the art can, in accordance with all the teachings disclosed, make various modifications and changes in the details, all of which are within the scope of the present invention, the full scope of which is set forth in the appended claims and any equivalents thereof.

Claims

1. Formula I, Formula II, or Formula III: 【Chemistry 1】 In, L is selected from the ligand; M is selected from any linker unit or combination; A is selected from any bridging scaffolding; B may or may not exist, and if present, it may be selected from any bridging structure or bond; C may or may not exist, and if it exists, it is selected from any branching unit or combination; D, D 1 , D 2 They are either identical or different, and each is independently selected from the drugs; SU is selected from sugars or their derivatives; L a and L b They may be identical or different, and each may be independently selected from any bridging unit or bond; m is selected from integers between 1 and 5; n is selected from integers between 1 and 10; o is selected from integers between 1 and 10. Ligand-drug conjugates, or pharmaceutically acceptable salts or solvates thereof.

2. The ligand-drug conjugate according to claim 1, or a pharmaceutically acceptable salt or solvate thereof, wherein the ligand L is selected from an antibody, a functional antibody fragment, and a protein having a targeting effect.

3. The ligand-drug conjugate according to claim 1, or a pharmaceutically acceptable salt or solvate thereof, wherein the ligand L is selected from, but is not limited to, an antibody comprising a chimeric antibody, a humanized antibody, a fully human antibody, or a mouse antibody.

4. The aforementioned linker unit M is given by the following formula: 【Chemistry 2】 The linker unit M is selected non-restrictively from the structures or bonds represented by; if the structure includes a cycloalkyl or heterocyclil, the linker unit M may also be selected from the derived structures in which the cycloalkyl or heterocyclil is ring-opened. During the ceremony, The positions indicated by the dashed lines are non-restrictively connected to the ligand, bridging scaffold A, bridging structure B, or branching unit C; * The carbon at the position indicated by is a chiral carbon and has either an R configuration or an S configuration; p and p' are each independently selected from integers between 1 and 10; Ac is a residue of a natural or unnatural amino acid, a polyethylene glycol segment having 1 to 20 repeating units, a phosphate group, a carboxylic acid group, a sulfonic acid group, a sulfinic acid group, or the following structure: 【Transformation 3】 Selected without restriction from, In the formula, the position indicated by the dashed line is, * The carbon atom at the position indicated by is linked to, The ligand-drug conjugate according to claim 1, or a pharmaceutically acceptable salt or solvate thereof.

5. The aforementioned linker unit M is given by the following formula: 【Chemistry 4】 A structure represented by, or a non-restrictive selection from these stereoisomers or derived structures in which the succinimide ring is in an open-ring form, During the ceremony, The positions indicated by the dashed lines are non-restrictively connected to the ligand, bridging scaffold A, bridging structure B, or branching unit C; * The carbon at the position indicated by is a chiral carbon and has either an R configuration or an S configuration; p and p' are each independently selected from integers between 1 and 10; Ac is a residue of a natural or unnatural amino acid, a polyethylene glycol segment having 1 to 20 repeating units, a phosphate group, a carboxylic acid group, a sulfonic acid group, a sulfinic acid group, or the following structure: 【Transformation 5】 Selected without restriction from, In the formula, the position indicated by the dashed line is, * The carbon atom at the position indicated by is linked to, The ligand-drug conjugate according to claim 1, or a pharmaceutically acceptable salt or solvate thereof.

6. The aforementioned linker unit M is given by the following formula: 【Transformation 6】 The structure represented by, or these derived structures in which the succinimide ring is in an open-ring form, preferably, 【Transformation 7】 Selected indefinitely from; During the ceremony, * The carbon at the position shown by is a chiral carbon and has an R configuration or an S configuration; The positions indicated by the dashed lines are not limited to being connected to the ligand, bridging scaffold A, bridging structure B, or branching unit C. The ligand-drug conjugate according to claim 1, or a pharmaceutically acceptable salt or solvate thereof.

7. The aforementioned cross-linking scaffold A is made of natural or unnatural amino acids or the following structure: 【Transformation 8】 One or more of the following are selected in an unrestricted manner: During the ceremony, X is selected from N, CH, C3-C8 cycloalkyl, 3- to 8-membered heterocyclyl, aryl, substituted aryl, and heteroaryl; Y is -NH-, -O-, -S-, -CO-, -CO 2 -, -CONH-, -NHCO-, -SO-, -SO 2 -, -OSO 2 - and -OP=O(OH)O- are selected; q is selected from integers between 1 and 10; The positions indicated by the dashed lines are linker unit M and bridge unit L. a Or L b , non-restrictively connected to bridge structure B or branching unit C; R 1 This is selected from hydrogen atoms, deuterium atoms, halogens, C1-C6 alkyl, C1-C6 substituted alkyl, C3-C8 cycloalkyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkyl, carboxyl, 3-8 membered heterocyclyl, aryl, substituted aryl, and heteroaryl; R 2 teeth, 【Chemistry 9】 Selected indefinitely from; In the formula, the wavy line on the left represents R 2 but * The hyphen indicates the position where the chiral carbon is connected, and the dashed line on the right indicates one of three arbitrary connection positions on the bridging scaffold A. The ligand-drug conjugate according to claim 1, or a pharmaceutically acceptable salt or solvate thereof.

8. The aforementioned bridging scaffold A has the following structure: 【Chemistry 10】 Or, non-restrictively selected from these stereoisomers, During the ceremony, Z is selected from -NH-, -O-, and -S-; The positions indicated by the dashed lines are linker unit M and bridge unit L. a Or L b , non-restrictively connected to bridge structure B or branching unit C; Preferably, the bridging scaffold A is 【Chemistry 11】 And in the formula, Z is -NH-. The ligand-drug conjugate according to claim 1, or a pharmaceutically acceptable salt or solvate thereof.

9. The ligand-drug complex according to claim 1, or a pharmaceutically acceptable salt or solvate thereof, wherein B is non-limitingly selected from any cross-linking structure or bond.

10. The ligand-drug complex according to claim 1, or a pharmaceutically acceptable salt or solvate thereof, wherein C is present or absent, and if present, is non-limitingly selected from one or more natural or non-natural amino acids.

11. The aforementioned drugs D, D 1 , and D 2 The ligand-drug conjugate or pharmaceutically acceptable salt or solvate thereof according to claim 1, wherein the elements are identical or different, and each is independently and non-limitingly selected from antitumor agents, autoimmune disease agents, anti-infective agents (e.g., antiviral agents), radioisotopes, chromogenic molecules, or pharmaceutically acceptable salts or solvates thereof.

12. The aforementioned drugs D, D 1 , and D 2 A ligand-drug conjugate according to claim 1, or a pharmaceutically acceptable salt or solvate thereof, which is identical or different from, but not limited to, an antitumor agent, including a DNA damaging agent, an RNA damaging agent, an enzyme inhibitor, or a microtubule inhibitor, selected independently and non-limitingly.

13. The ligand-drug conjugate according to claim 1, or a pharmaceutically acceptable salt or solvate thereof, wherein the SU is non-limitingly selected from natural or unnatural monosaccharides, disaccharides, polysaccharides and derivatives thereof.

14. The aforementioned natural or unnatural monosaccharides, disaccharides, polysaccharides, or derivatives thereof have the following structure: 【Chemistry 12-1】 【Chemistry 12-2】 【Chemistry 12-3】 Selected without restriction from, Preferably, the SU is selected from methylglucamine, maltose, maltobionic acid, mannuronic acid, β-cyclodextrin, and mono(6-amino-6-deoxy)-β-cyclodextrin. The ligand-drug conjugate according to claim 1, or a pharmaceutically acceptable salt or solvate thereof.

15. The SU is covalently bonded to L b A ligand-drug complex according to claim 1, or a pharmaceutically acceptable salt or solvate thereof, non-limitingly linked to the ligand-drug complex.

16. The aforementioned cross-linking unit L a and L b The ligand-drug complex according to claim 1, or a pharmaceutically acceptable salt or solvate thereof, wherein the units are identical or different, and each is independently and non-limitingly selected from one or more chemically unstable crosslinking units, enzyme-catalyzed cleavable crosslinking units, and non-cleavable crosslinking units.

17. L a The ligand-drug complex according to claim 1, or a pharmaceutically acceptable salt or solvate thereof, wherein the crosslinking units are selected from chemically unstable crosslinking units and enzyme-catalyzed crosslinking units.

18. The aforementioned cross-linking unit L a The structure is as follows: 【Chemistry 13】 Or, non-restrictively selected from these stereoisomers, During the ceremony, R a , R b , and R c They are either identical or different, and each is independently selected from hydrogen atoms, deuterium atoms, halogens, alkyls, substituted alkyls, deuterated alkyls, cycloalkyls, cycloalkylalkyls, alkoxyalkyls, heterocyclyls, aryls, substituted aryls, or heteroaryls; Or, R b , R c and R b and R c The carbon atoms linked thereto constitute a C3-C8 cycloalkyl or a 3-8 membered heterocycline; R d H, NO 2 HO-SO 3 --and --OSO 3 Selected from; r is selected from integers between 1 and 10; The location indicated by the wavy line on the left is connected to bridge scaffolding A, bridge structure B, or branching unit C; The location indicated by the wavy line on the right is drug D, D 1 Or D 2 It is connected to, The ligand-drug conjugate according to claim 1, or a pharmaceutically acceptable salt or solvate thereof.

19. The aforementioned cross-linking unit L a -D, L a -D 1 and L a -D 2 They are either identical or different, and have the following structure: (1) Auristatin 【Chemistry 14】 (2) Maytansinoids 【Chemistry 15】 (3) Benzodiazepines 【Chemistry 16】 (4) Camptothecin analogs 【Chemistry 17】 (5) Tubericin [Chemistry 18] (6) Adriamycin 【Chemistry 19】 (7) Calicheamycin 【Chemistry 20】 (8) Duocalmycin 【Chemistry 21】 (9) Other antitumor drugs 【Chemistry 22-1】 【Chemistry 22-2】 Or they are selected non-restrictively from these stereoisomers; During the ceremony, The positions indicated by the dashed lines are connected to bridge scaffolding A, bridge structure B, or branching unit C; Preferably, the L a -D, L a -D 1 or L a -D 2 teeth, Auristatin, for example, 【Chemistry 23】 , benzodiazepines, for example, 【Chemistry 24】 , camptothecin analogs, for example, 【Chemistry 25】 Combretastatin, for example, 【Chemistry 26】 A ligand-drug conjugate according to claim 1, or a pharmaceutically acceptable salt or solvate thereof, selected from the above.

20. The aforementioned cross-linking unit L b The structure is as follows: 【Chemistry 27】 Or, non-restrictively selected from these stereoisomers, During the ceremony, R 3 This is selected from hydrogen atoms, deuterium atoms, halogens, C1-C6 alkyl, deuterated C1-C6 alkyl, halogenated C1-C6 alkyl, C3-C8 cycloalkyl C1-C6 alkyl, C1-C6 alkoxy C1-C6 alkyl, heterocyclyl, aryl, substituted aryl, and heteroaryl; W is -NR 4 -, -O-, -S-, -CO-, and -CONR 5 - Selected from; R 4 or R 5 These are independently selected from hydrogen atoms, deuterium atoms, halogens, C1-C6 alkyls, deuterated C1-C6 alkyls, halogenated C1-C6 alkyls, C3-C8 cycloalkyls, C1-C6 alkoxys, heterocyclyls, aryls, substituted aryls, and heteroaryls; s is selected from integers between 1 and 10; s' is selected from integers between 1 and 10; The positions indicated by the left and right wavy lines are not limited to being connected to the SU or bridge scaffolding A. The ligand-drug conjugate according to claim 1, or a pharmaceutically acceptable salt or solvate thereof.

21. Formulas IV, V, or VI: 【Chemistry 28】 In, M is selected from any linker unit; A is selected from any bridging scaffolding; B may or may not exist, and if present, it may be selected from any bridging structure or bond; C may or may not exist, and if it exists, it is selected from any branching unit or combination; D, D 1 , D 2 They are either identical or different, and each is independently selected from the drugs; SU is selected from sugars or their derivatives; L a and L b They may be identical or different, and each may be independently selected from any bridging unit or bond; m is selected from integers between 1 and 5; o is selected from integers between 1 and 10. Linker drug compounds or their pharmaceutically acceptable salts or solvates.

22. The aforementioned linker unit M is given by the following formula: 【Chemistry 29】 The structure is not limited to those represented by; if the structure includes a cycloalkyl or heterocyclyl, the linker unit M may also be selected from its derived structure in which the cycloalkyl or heterocyclyl is in an open-ring form. During the ceremony, The positions indicated by the dashed lines are not limited to being connected to bridge scaffolding A, bridge structure B, or branching unit C; M' is non-restrictively selected from halogen, OTf, and OTs; * The carbon at the position indicated by is a chiral carbon and has either an R configuration or an S configuration; p and p' are each independently selected from integers between 1 and 10; Ac is a residue of a natural or unnatural amino acid, a polyethylene glycol segment having 1 to 20 repeating units, a phosphate group, a carboxylic acid group, a sulfonic acid group, a sulfinic acid group, or the following structure: 【Transformation 30】 Selected without restriction from, In the formula, the position indicated by the dashed line is, * The carbon atom at the position indicated by is linked to, The linker drug compound according to claim 21, or a pharmaceutically acceptable salt or solvate thereof.

23. The aforementioned linker unit M is given by the following formula: 【Chemistry 31】 A structure represented by, or a non-restrictive selection from these stereoisomers or derived structures in which the succinimide ring is in an open-ring form, During the ceremony, The positions indicated by the dashed lines are not limited to being connected to bridge scaffolding A, bridge structure B, or branching unit C; * The carbon at the position indicated by is a chiral carbon and has either an R configuration or an S configuration; p and p' are each independently selected from integers between 1 and 10; Ac is a residue of a natural or unnatural amino acid, a polyethylene glycol segment having 1 to 20 repeating units, a phosphate group, a carboxylic acid group, a sulfonic acid group, a sulfinic acid group, or the following structure: 【Chemistry 32】 Selected without restriction from, In the formula, the position indicated by the dashed line is, * The carbon atom at the position indicated by is linked to, The linker drug compound according to claim 21, or a pharmaceutically acceptable salt or solvate thereof.

24. The aforementioned linker unit M is given by the following formula: 【Transformation 33】 A structure represented by, or any derived structure in which the succinimide ring is in an open-ring form, can be selected non-restrictively from these structures. Preferably, 【Transformation 34】 Selected from, in the formula, * The carbon atom at the position indicated by is a chiral carbon and has either an R configuration or an S configuration; The positions indicated by the dashed lines are not limited to being connected to bridge scaffolding A, bridge structure B, or branching unit C. The linker drug compound according to claim 21, or a pharmaceutically acceptable salt or solvate thereof.

25. The aforementioned cross-linking scaffold A is made of natural or unnatural amino acids or the following structure: 【Chemistry 35】 One or more of the following are selected: During the ceremony, X is selected from N, CH, C3-C8 cycloalkyl, 3- to 8-membered heterocyclyl, aryl, substituted aryl, and heteroaryl; Y is -NH-, -O-, -S-, -CO-, -CO 2 -, -CONH-, -NHCO-, -SO-, -SO 2 -, -OSO 2 - and -OP=O(OH)O- are selected; q is selected from integers between 1 and 10; The positions indicated by the dashed lines are linker unit M and bridge unit L. a Or L b , non-restrictively connected to bridge structure B or branching unit C; R 1 This is selected from hydrogen atoms, deuterium atoms, halogens, C1-C6 alkyl, C1-C6 substituted alkyl, C3-C8 cycloalkyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkyl, carboxyl, 3-8 membered heterocyclyl, aryl, substituted aryl, and heteroaryl; R 2 teeth, 【Transformation 36】 Selected without restriction from, In the formula, the wavy line on the left represents R 2 but * The hyphen indicates the position where the chiral carbon is connected, and the dashed line on the right indicates one of three arbitrary connection positions on the bridging scaffold A. The linker drug compound according to claim 21, or a pharmaceutically acceptable salt or solvate thereof.

26. The aforementioned bridging scaffold A has the following structure: 【Chemistry 37】 Or, non-restrictively selected from these stereoisomers, During the ceremony, Z is selected from -NH-, -O-, and -S-; The positions indicated by the dashed lines are linker unit M and bridge unit L. a Or L b , non-restrictively connected to bridge structure B or branching unit C; Preferably, the bridging scaffold A is 【Transformation 38】 And in the formula, Z is -NH-. The linker drug compound according to claim 21, or a pharmaceutically acceptable salt or solvate thereof.

27. The linker drug compound according to claim 21, or a pharmaceutically acceptable salt or solvate thereof, wherein B is non-limitingly selected from any cross-linking structure or bond.

28. The linker drug compound according to claim 21 or a pharmaceutically acceptable salt or solvate thereof, wherein C is present or absent, and if present, is non-limitingly selected from one or more natural or unnatural amino acids.

29. The aforementioned drugs D, D 1 , and D 2 The linker drug compound according to claim 21 or a pharmaceutically acceptable salt or solvate thereof, which is identical or different and independently and non-limitingly selected from antitumor drugs, autoimmune disease drugs, anti-infective drugs (e.g., antiviral drugs), radioisotopes, chromogenic molecules, or pharmaceutically acceptable salts or solvates thereof.

30. The aforementioned drugs D, D 1 , and D 2 The linker drug compound according to claim 21, or a pharmaceutically acceptable salt or solvate thereof, which is identical or different from, but not limited to, an antitumor agent, which includes a DNA damaging agent, an RNA damaging agent, an enzyme inhibitor, or a microtubule inhibitor.

31. The linker drug compound according to claim 21, or a pharmaceutically acceptable salt or solvate thereof, wherein the SU is not limited to being selected from natural or unnatural monosaccharides, disaccharides, polysaccharides and derivatives thereof.

32. The aforementioned natural or unnatural monosaccharides, disaccharides, polysaccharides, or derivatives thereof have the following structure: 【Chemistry 39-1】 【Chemistry 39-2】 【Chemistry 39-3】 Selected indefinitely from; Preferably, the SU is selected from methylglucamine, maltose, maltobionic acid, mannuronic acid, β-cyclodextrin, and mono(6-amino-6-deoxy)-β-cyclodextrin. The linker drug compound according to claim 21, or a pharmaceutically acceptable salt or solvate thereof.

33. The SU is covalently bonded to L b A linker drug compound according to claim 21 or a pharmaceutically acceptable salt or solvate thereof, which is non-limitingly linked to the linker drug compound according to claim 21.

34. The aforementioned cross-linking unit L a and L b The linker drug compound according to claim 21 or a pharmaceutically acceptable salt or solvate thereof, wherein the linker is selected from one or more of the following: identical or different, independently and non-limitingly chemically unstable crosslinking units, enzyme-catalyzed cleavable crosslinking units, and non-cleavable crosslinking units.

35. L a The linker drug compound according to claim 21, or a pharmaceutically acceptable salt or solvate thereof, wherein the linker is selected from chemically unstable crosslinking units and enzyme-catalyzed crosslinking units.

36. The aforementioned cross-linking unit L a The structure is as follows: 【Chemistry 40】 Or, non-restrictively selected from these stereoisomers, During the ceremony, R a 、 R b 、 and R c are the same or different and each independently is selected from a hydrogen atom, a deuterium atom, a halogen, an alkyl, a substituted alkyl, a deuterated alkyl, a cycloalkyl, a cycloalkylalkyl, an alkoxyalkyl, a heterocyclyl, an aryl, a substituted aryl or a heteroaryl; Or, R b , R c and R b and R c The carbon atoms linked thereto constitute a C3-C8 cycloalkyl or a 3-8 membered heterocycline; R d is selected from H, NO 2 , HO-SO 3 and -OSO 3 ; and r is selected from integers between 1 and 10; The location indicated by the wavy line on the left is connected to bridge scaffolding A, bridge structure B, or branching unit C; The location indicated by the wavy line on the right is drug D, D 1 Or D 2 It is connected to, The linker drug compound according to claim 21, or a pharmaceutically acceptable salt or solvate thereof.

37. Said L a -D, L a -D 1 and L a -D 2 They are either identical or different, and have the following structure: (1) Auristatin 【Chemistry 41】 (2) Maytansinoids 【Chemistry 42】 (3) Benzodiazepines 【Chemistry 43】 (4) Camptothecin analogs 【Chemistry 44】 (5) Tubericin 【Chemistry 45】 (6) Adriamycin 【Chemistry 46】 (7) Calicheamycin 【Chemistry 47】 (8) Duocalmycin 【Chemistry 48】 (9) Other antitumor drugs 【Chemistry 49】 Or they are selected non-restrictively from these stereoisomers; During the ceremony, The positions indicated by the dashed lines are connected to bridge scaffolding A, bridge structure B, or branching unit C; Preferably, the L a -D, L a -D 1 or L a -D 2 teeth, Auristatin, for example, [Transformation 50] , benzodiazepines, for example, 【Chemistry 51】 , camptothecin analogs, for example, 【Chemistry 52】 Combretastatin, for example, 【Chemistry 53】 Selected from, The linker drug compound according to claim 21, or a pharmaceutically acceptable salt or solvate thereof.

38. The aforementioned cross-linking unit L b The structure is as follows: 【Chemistry 54】 Or, non-restrictively selected from these stereoisomers, During the ceremony, R 3 This is selected from hydrogen atoms, deuterium atoms, halogens, C1-C6 alkyl, deuterated C1-C6 alkyl, halogenated C1-C6 alkyl, C3-C8 cycloalkyl C1-C6 alkyl, C1-C6 alkoxy C1-C6 alkyl, heterocyclyl, aryl, substituted aryl, and heteroaryl; W is -NR 4 -, -O-, -S-, -CO-, and -CONR 5 - Selected from; R 4 or R 5 These are independently selected from hydrogen atoms, deuterium atoms, halogens, C1-C6 alkyls, deuterated C1-C6 alkyls, halogenated C1-C6 alkyls, C3-C8 cycloalkyls, C1-C6 alkoxys, heterocyclyls, aryls, substituted aryls, and heteroaryls; s is selected from integers between 1 and 10; s' is selected from integers between 1 and 10; The positions indicated by the left and right wavy lines are not limited to being connected to the SU or bridge scaffolding A. The linker drug compound according to claim 21, or a pharmaceutically acceptable salt or solvate thereof.

39. Said L b -SU has the following structure: 【Chemistry 55-1】 【Chemistry 55-2】 【Chemistry 55-3】 【Chemistry 55-4】 【Transformation 55-5】 【Chemistry 55-6】 【Chemistry 55-7】 A linker drug compound according to claim 21, or a pharmaceutically acceptable salt or solvate thereof, non-limitingly selected from the above.

40. The aforementioned linker drug compound has the following structure: 【Chemistry 56-1】 【Chemistry 56-2】 【Chemistry 56-3】 【Chemistry 56-4】 【Chemistry 56-5】 A linker drug compound according to claim 21, or a pharmaceutically acceptable salt or solvate thereof, non-limitingly selected from the above.

41. The aforementioned linker drug compound has the following structure: 【Chemistry 57-1】 【Chemistry 57-2】 【Chemistry 57-3】 A linker drug compound according to claim 21, or a pharmaceutically acceptable salt or solvate thereof, non-limitingly selected from the above.

42. The aforementioned linker drug compound has the following structure: 【Chemistry 58-1】 【Chemistry 58-2】 【Chemistry 58-3】 A linker drug compound according to claim 21, or a pharmaceutically acceptable salt or solvate thereof, non-limitingly selected from the above.

43. The aforementioned linker drug compound has the following structure: 【Chemistry 59-1】 【Chemistry 59-2】 【Chemistry 59-3】 A linker drug compound according to claim 21, or a pharmaceutically acceptable salt or solvate thereof, non-limitingly selected from the above.

44. A method for preparing a ligand-drug conjugate of formula I, formula II, or formula III as described in claim 1, or a pharmaceutically acceptable salt or solvate thereof, comprising the following steps: 【Transformation 60】 The process includes reacting a modified ligand with a compound of formula IV, V, or VI to obtain a ligand-drug conjugate of formula I, II, or III, or a pharmaceutically acceptable salt or solvate thereof. During the ceremony, L is selected from the ligand; M is selected from any linker unit; A is selected from any bridging scaffolding; B may or may not exist, and if present, it may be selected from any bridging structure or bond; C may or may not exist, and if it exists, it is selected from any branching unit or combination; D, D 1 , D 2 They are either identical or different, and each is independently selected from the drugs; SU is selected from sugars or their derivatives; L a and L b They may be identical or different, and each may be independently selected from any bridging unit or bond; m is selected from integers between 1 and 5; n is selected from integers between 1 and 10; o is selected from integers between 1 and 10. method.

45. A ligand-drug conjugate or a pharmaceutically acceptable salt or solvate thereof has the following structure: 【Chemistry 61-1】 【Chemistry 61-2】 【Chemistry 61-3】 【Chemistry 61-4】 【Chemistry 61-5】 【Chemistry 61-6】 【Chemistry 61-7】 【Chemistry 61-8】 【Chemistry 61-9】 【Chemistry 61-10】 【Chemistry 61-11】 【Chemistry 61-12】 【Chemistry 61-13】 Or, non-restrictively selected from these stereoisomers, During the ceremony, L is selected from a ligand, preferably an antibody; n is selected from integers 1 to 10, preferably 2 to 8. The ligand-drug conjugate according to claim 1, or a pharmaceutically acceptable salt or solvate thereof.

46. The aforementioned pharmaceutically acceptable salt is Sodium salt, potassium salt, calcium salt, or magnesium salt formed by the acidic functional group in the aforementioned structural formula and an alkali; and Acetates, trifluoroacetates, citrates, oxalates, tartrates, malates, nitrates, chlorides, bromides, iodides, sulfates, bisulfates, phosphates, lactates, oleates, ascorbic acid, salicylates, formates, glutamates, methanesulfonates, ethanesulfonates, benzenesulfonates, or p-toluenesulfonates formed by the basic functional groups in the aforementioned structure and acids. A ligand-drug conjugate according to claim 1, or a pharmaceutically acceptable salt thereof, selected from the above.

47. The ligand is selected from monoclonal antibodies, including anti-EGFRvIII antibody, anti-DLL-3 antibody, anti-PSMA antibody, anti-CD70 antibody, anti-MUC16 antibody, anti-ENPP3 antibody, anti-TDGF1 antibody, anti-ETBR antibody, anti-MSLN antibody, anti-TIM-1 antibody, anti-LRRC15 antibody, anti-LIV-1 antibody, anti-CanAg / AFP antibody, anti-claudin 18.2 antibody, anti-mesothelin antibody, anti-HER2 ( ErbB2) antibody, anti-EGFR antibody, anti-C-Met antibody, anti-SLITRK6 antibody, anti-KIT / CD117 antibody, anti-STEAP1 antibody, anti-SLAMF7 / CS1 antibody, anti-NaPi2B / SLC34 A2 antibody, anti-GPNMB antibody, anti-HER3 (ErbB3) antibody, anti-MUC1 / CD227 antibody, anti-AXL antibody, anti-CD166 antibody, anti-B7-H3 (CD276) antibody, anti-PTK7 / CCK4 antibody , anti-PRLR antibody, anti-EFNA4 antibody, anti-5T4 antibody, anti-NOTCH3 antibody, anti-Nectin-4 antibody, anti-Trop-2 antibody, anti-CD142 antibody, anti-CA6 antibody, anti-GPR20 antibody, anti-CD174 antibody body, anti-CD71 antibody, anti-EphA2 antibody, anti-LYPD3 antibody, anti-FGFR2 antibody, anti-FGFR3 antibody, anti-FRα antibody, anti-CEACAMs antibody, anti-GCC antibody, anti-integrin Av antibody, anti-CAI Non-limiting selection from X antibody, anti-P-cadherin antibody, anti-GD3 antibody, anti-cadherin 6 antibody, anti-LAMP1 antibody, anti-FLT3 antibody, anti-BCMA antibody, anti-CD79b antibody, anti-CD19 antibody, anti-CD33 antibody, anti-CD56 antibody body, anti-CD74 antibody, anti-CD22 antibody, anti-CD30 antibody, anti-CD37 antibody, anti-CD138 antibody, anti-CD352 antibody, anti-CD25 antibody, and anti-CD123 antibody; Preferably, the ligand is an anti-Trop-2 antibody or an anti-CD33 antibody. The ligand-drug conjugate or pharmaceutically acceptable salt according to claim 1.

48. A pharmaceutical composition comprising a therapeutically effective amount of a ligand-drug complex or a pharmaceutically acceptable salt of a ligand-drug complex according to any one of claims 1 to 38 and 45 to 47, or a linker drug compound or a pharmaceutically acceptable salt or solvate of a linker drug compound according to any one of claims 21 to 43, and a pharmaceutically acceptable carrier, diluent, or excipient.

49. Use of a ligand-drug conjugate or a pharmaceutically acceptable salt of a ligand-drug conjugate according to any one of claims 1 to 38 and 45 to 47, or a linker drug compound or a pharmaceutically acceptable salt or solvate of a linker drug compound according to any one of claims 21 to 43, in the preparation of a drug for treating or preventing a tumor or autoimmune disease.

50. The use according to claim 49, wherein the tumor is selected from solid tumors or non-solid tumors, for example, breast cancer, ovarian cancer, cervical cancer, uterine cancer, prostate cancer, kidney cancer, urethral cancer, bladder cancer, liver cancer, stomach cancer, endometrial cancer, salivary gland cancer, esophageal cancer, lung cancer, colon cancer, rectal cancer, colorectal cancer, bone cancer, skin cancer, thyroid cancer, pancreatic cancer, melanoma, glioma, neuroblastoma, glioblastoma multiforme, sarcoma, lymphoma, and leukemia.

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