Iproxetecan derivative as well as preparation method and application thereof

Abstract

The invention relates to the field of biological medicines, and particularly relates to an etiotecan derivative as well as a preparation method and application thereof. The relaxetecan derivative is specifically as shown in a formula (I). The definition of each group in the formula (I) is shown in the specification. The compound provided by the invention has relatively good anti-tumor activity.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, specifically to ixotecan derivatives, their preparation methods, and applications. Background Technology

[0002] Antibody-drug conjugates (ADCs) deliver cytotoxic drugs (or "toxins") to tumor cells via antibody carriers. Toxins play a crucial role in ADCs; they act as the direct cytotoxic agents, exerting a powerful killing effect on tumor cells. Currently, the cytotoxic payloads in ADCs mainly include potent microtubule inhibitors, DNA damaging agents, and immunomodulators. These toxins share the common characteristics of remaining stable under physiological conditions and possessing functional groups that can covalently bind to antibodies.

[0003] The selection and design of toxins are crucial to the efficacy and safety of ADCs. They need to possess sufficient toxicity to effectively kill cancer cells after payload release, while remaining stable in the bloodstream to avoid damage to normal cells. Furthermore, the chemical and physical properties of the toxins also affect the overall pharmacokinetic and pharmacodynamic characteristics of the ADC.

[0004] In recent years, most clinical trials and marketed antibody-drug conjugates (ADCs) have used ecstasylate (DX-8951f), a camptothecin derivative and a lower-potency but safer inhibitor of DNA topoisomerase I (TOP1), as the toxin. Ecitabine, as an ADC payload, has several significant advantages, such as potent killing ability, rapid rate of action, and minimal impact on multidrug resistance (MDR) mechanisms. However, ecstasylate itself has strong toxicity, particularly gastrointestinal and myelotoxicity. This limits its clinical application as a standalone small molecule drug, as high toxicity may lead to patient intolerance, limiting dosage and therapeutic window. Furthermore, ecstasylate's high hydrophobicity makes it difficult to directly conjugate with antibodies. Excessive hydrophobicity may lead to stability and solubility issues in the ADC in vivo, affecting the drug's pharmacokinetic properties and efficacy.

[0005] Given the challenges faced by eczemac in the development of antibody-drug conjugates, it is particularly urgent and important to develop an eczemac derivative with lower toxicity and better anti-tumor efficacy. Summary of the Invention

[0006] In light of the aforementioned problems, this disclosure aims to develop more eczema derivatives with good anti-tumor efficacy and low toxicity. The compounds provided in this disclosure possess excellent tumor cell inhibitory activity and can be used as monotherapy, in combination therapy, or as a toxic component of antitumor agonists (ADCs) for cancer treatment.

[0007] In one aspect of this disclosure, compounds of formula (I), pharmaceutically acceptable salts thereof, stereoisomers thereof, solvates or prodrugs are provided.

[0008]

[0009] In the formula,

[0010] R1 and R2 are each independently selected from hydrogen, fluorine, chlorine, amino, nitro, hydroxyl, and methyl.

[0011] R3 is selected from -CR4R5-X1R6R7, -X2R8R9, or R 10 ;

[0012] X1 is selected from N or CR 11 ;

[0013] X2 is selected from N or CR 12 ;

[0014] R4 and R5 are each independently selected from hydrogen, halogen, hydroxyl, substituted or unsubstituted C. 1-6 Alkyl and substituted or unsubstituted C 1-6 Alkoxy;

[0015] Alternatively, R4, R5, and the carbon atoms they are attached to can form substituted or unsubstituted C atoms. 3-10 Cycloalkyl and substituted or unsubstituted 3- to 10-membered heterocyclic alkyl groups;

[0016] R6 and R7 are each independently selected from hydrogen, halogen, oxo group, hydroxyl group, substituted or unsubstituted C. 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 3-10 Cycloalkyl groups and substituted or unsubstituted 3- to 10-membered nitrogen-containing heterocyclic alkyl groups;

[0017] Alternatively, R6, R7, and the carbon or nitrogen atom they are attached to form substituted or unsubstituted C atoms. 3-10 Cycloalkyl or substituted or unsubstituted 3- to 10-membered heterocyclic alkyl groups;

[0018] R8 is selected from substituted or unsubstituted C. 3-10 Cycloalkyl and substituted or unsubstituted 3- to 10-membered heterocyclic alkyl groups;

[0019] R9 is selected from hydrogen, halogen, hydroxyl, substituted or unsubstituted C. 1-6Alkyl and substituted or unsubstituted C 1-6 Alkoxy;

[0020] R 10 Selected from substituted or unsubstituted C 3-10 Cycloalkyl and substituted or unsubstituted 3- to 10-membered heterocyclic alkyl groups;

[0021] R 11 R 12 Selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl and halogen-substituted C 1-6 alkyl;

[0022] The term "substitution" as used herein refers to each of the aforementioned groups being independently replaced by one or more groups selected from R. A The substituents are replaced by the R; A Selected from hydrogen, deuterium, halogen, hydroxyl, oxo, cyano, nitro, C 1-6 Alkyl, C 1-6 Alkoxy group, -NHC(O)-C 1-6 Alkyl group, -NHS(O)2-C 1-6 Alkyl, C 3-10 Cycloalkyl and 3 to 10-membered heterocycloalkyl, wherein C 1-6 Alkyl, C 1-6 Alkoxy group, -NHC(O)-C 1-6 Alkyl group, -NHS(O)2-C 1-6 Alkyl, C 3-10 The cycloalkyl or 3- to 10-membered heterocycloalkyl group is optionally substituted by one or more substituents selected from halogens and hydroxyl groups;

[0023] In each of the above groups, the 3 to 10-membered heterocyclic alkyl group may optionally contain 1, 2, 3 or 4 heteroatoms selected from nitrogen, oxygen or sulfur as ring atoms, and the ring atoms of the 3 to 10-membered nitrogen-containing heterocyclic alkyl group may contain at least 1 nitrogen atom and optionally also contain 1, 2 or 3 heteroatoms selected from nitrogen, oxygen or sulfur as ring atoms.

[0024] In some implementations, R3 contains at least one hydroxyl substituent.

[0025] In some implementations, R1 and R2 are each independently selected from hydrogen, fluorine, methyl, and amino.

[0026] In some specific implementations, R1 is methyl and R2 is fluorine; or R1 is amino and R2 is hydrogen.

[0027] In some specific implementations, R1 is methyl and R2 is fluorine.

[0028] In some implementation schemes, R4 and R5 are each independently selected from hydrogen, halogen, hydroxyl, and C. 1-6Alkyl and C 1-6 Alkoxy, the C 1-6 Alkyl and C 1-6 The alkoxy group is optionally surrounded by 1, 2, 3 or 4 groups selected from R A Substituents are substituted.

[0029] In some specific implementation schemes, R4 and R5 are each independently selected from hydrogen, fluorine, and chlorine.

[0030] In some specific implementations, R4 and R5 are each independently selected from hydrogen and fluorine.

[0031] In some specific implementation schemes, R4 and R5 are both hydrogen or both R4 and R5 are fluorine.

[0032] In some implementations, R4, R5, and the carbon atoms they are bonded to together form C. 3-10 cycloalkyl, the C 3-10 The cycloalkyl group is optionally surrounded by 1, 2, 3 or 4 groups selected from R A Substituents are substituted.

[0033] In some specific implementations, R4, R5, together with the carbon atoms they are attached to, form cyclopropyl, cyclobutyl, cycloalkyl, or cyclohexyl groups.

[0034] In some specific implementations, R4, R5 and the carbon atoms they are attached to together form cyclopropyl or cyclobutyl.

[0035] In some implementation schemes, R6 and R7 are each independently selected from hydrogen, halogen, hydroxyl, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 3-10 Cycloalkyl and 3 to 10-membered heterocycloalkyl, wherein C 1-6 Alkyl, C 1-6 Alkoxy, C 3-10 Cycloalkyl or 3 to 10-membered heterocycloalkyl groups optionally composed of 1, 2, 3 or 4 selected from R A Substituents are substituted.

[0036] In some specific implementation schemes, R6 and R7 are each independently selected from hydrogen, halogen, hydroxyl, and C. 1-6 Alkyl and halogen-substituted C 1-6 alkyl.

[0037] In some specific implementation schemes, R6 and R7 are each independently selected from hydrogen, halogen, hydroxyl, and C. 1-3 Alkyl and halogen-substituted C 1-3 alkyl.

[0038] In some specific implementations, R6 and R7 are each independently selected from hydrogen, fluorine, chlorine, hydroxyl, methyl, ethyl, propyl, fluoromethyl, difluoromethyl, trifluoromethyl, fluoroethyl, difluoroethyl and trifluoroethyl.

[0039] In some specific implementations, R6 and R7 are each independently selected from hydroxyl, methyl, ethyl and trifluoromethyl.

[0040] In some specific implementation schemes, X1 is CR 11 ;R 11 Selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl and halogen-substituted C 1-6 alkyl.

[0041] In some specific implementation schemes, R 11 Selected from hydrogen and C 1-6 alkyl.

[0042] In some specific implementation schemes, R 11 Selected from hydrogen and C 1-3 alkyl.

[0043] In some specific implementation schemes, R 11 Selected from hydrogen and methyl.

[0044] In some implementations, X1 is CH or N; R6, R7, and the carbon or nitrogen atoms they are bonded to together form C. 3-10 Cycloalkyl or 3 to 10-membered heterocycloalkyl, wherein the C 3-10 Cycloalkyl or 3 to 10-membered heterocycloalkyl groups optionally composed of 1, 2, 3 or 4 selected from R A Substituents are substituted.

[0045] In some specific embodiments, R6, R7 and the carbon or nitrogen atom connected to them together form a 3 to 10-membered nitrogen-containing heterocyclic alkyl group, wherein the 3 to 10-membered nitrogen-containing heterocyclic alkyl group is optionally substituted by 1, 2, 3 or 4 substituents selected from hydroxyl or oxo groups.

[0046] In some implementations, R8 is selected from C 3-10 Cycloalkyl and substituted 3- to 10-membered heterocycloalkyl, wherein C 3-10 Cycloalkyl or 3 to 10-membered heterocycloalkyl groups optionally composed of 1, 2, 3 or 4 selected from R A Substituents are substituted.

[0047] In some specific implementation schemes, R8 is The Optionally selected from 1, 2, 3 or 4 of R A Substituents are substituted.

[0048] In some specific implementation schemes, R8 is The Optionally, it is selected from 1, 2, 3 or 4 halogens or C 1-6 Substitution of alkoxy groups.

[0049] In some specific implementation schemes, R8 is The Optionally, it is selected from 1, 2, 3 or 4 halogens or C 1-3 Substitution of alkoxy groups.

[0050] In some specific implementation schemes, R8 is The It may optionally be substituted with 1, 2, 3 or 4 substituents selected from fluorine or methoxy groups.

[0051] In some specific implementations, R9 is selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl and C 1-6 Alkyl-OH.

[0052] In some specific embodiments, R9 is selected from hydroxyl and C. 1-6 Alkyl-OH.

[0053] In some specific embodiments, R9 is selected from hydroxyl and C. 1-3 Alkyl-OH.

[0054] In some specific implementations, R9 is selected from hydroxyl and -(CH2)2-OH.

[0055] In some implementation schemes, R 10 Selected from C 3-10 Cycloalkyl and 3 to 10-membered heterocycloalkyl, wherein C 3-10 Cycloalkyl or 3 to 10-membered heterocycloalkyl groups optionally composed of 1, 2, 3 or 4 selected from R A Substituents are substituted.

[0056] In some specific implementation schemes, R 10 Selected from C 3-10 cycloalkyl, the C 3-10 The cycloalkyl group is optionally surrounded by one or two hydroxyl groups and -NHC(O)-C. 1-6 Substitution of alkyl-OH groups.

[0057] In some specific implementation schemes, R 10 Selected from cyclopropyl, cyclobutyl, cyclopentyl, and adamantyl, wherein the cyclopropyl, cyclobutyl, cyclopentyl, or adamantyl group is optionally surrounded by one or two groups selected from hydroxyl and -NHC(O)-C. 1-6 Substitution of alkyl-OH groups.

[0058] In some specific implementation schemes, R10 Selected from cyclopropyl, cyclobutyl, cyclopentyl, and adamantyl, wherein the cyclopropyl, cyclobutyl, cyclopentyl, or adamantyl group is optionally surrounded by one or two groups selected from hydroxyl and -NHC(O)-C. 1-3 Substitution of alkyl-OH groups.

[0059] In some specific implementation schemes, R 10 Selected from cyclopropyl, cyclobutyl, cyclopentyl, and adamantyl, wherein the cyclopropyl, cyclobutyl, cyclopentyl, or adamantyl group is optionally surrounded by one or two groups selected from hydroxyl and -NHC(O)-C. 1-3 Substitution of alkyl-OH groups.

[0060] In some specific implementation schemes, R 10 Selected from cyclobutyl and adamantyl, wherein the cyclobutyl or adamantyl group is optionally substituted with a substituent selected from hydroxyl or -NHC(O)-(CH2)2-OH.

[0061] In some implementation schemes, R 10 The derivatives are selected from aziridine, pyrrolidinyl, piperidinyl, and piperazine, wherein the aziridine, pyrrolidinyl, piperidinyl, or piperazine is optionally composed of 1, 2, 3, or 4 derivatives selected from R. A Substituents are substituted.

[0062] In some specific implementation schemes, R 10 The group is selected from aziridine, pyrrolidinyl, piperidinyl, and piperazine, wherein the aziridine, pyrrolidinyl, piperidinyl, or piperazine is optionally surrounded by 1, 2, 3, or 4 groups selected from -C. 1-6 Alkoxy and -C 1-6 Substitution of alkyl-OH groups.

[0063] In some specific implementation schemes, R 10 The group is selected from aziridine, pyrrolidinyl, piperidinyl, and piperazine, wherein the aziridine, pyrrolidinyl, piperidinyl, or piperazine is optionally surrounded by 1, 2, 3, or 4 groups selected from -C. 1-3 Alkoxy and -C 1-3 Substitution of alkyl-OH groups.

[0064] In some specific implementation schemes, R 10 Selected from aziridine and pyrrolidinyl groups, wherein the aziridine or pyrrolidinyl group is optionally surrounded by 1, 2, 3 or 4 groups selected from -C 1-3 Alkoxy and -C 1-3 Substitution of alkyl-OH groups.

[0065] In some specific implementation schemes, R 10 Selected from The Optionally substituted with one or two substituents selected from methoxy and -(CH2)2-OH.

[0066] In some implementations, the C 3-10 Each cycloalkyl group is independently selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, bicyclo[1.1.0]butyl, bicyclo[1.1.1]pentyl, bicyclo[2.2.0]hexyl, bicyclo[2.1.1]hexyl, bicyclo[3.1.0]hexyl, bicyclo[2.2.1]heptyl, bicyclo[3.2.0]heptyl, bicyclo[3.2.1]octyl, bicyclo[2.2.2]octyl, bicyclo[4.2.0]octyl, bicyclo[4.3.0]nonyl, bicyclo[3.3.1]nonyl, and adamantyl.

[0067] In some embodiments, the 3 to 10-membered heterocyclic alkyl groups are each independently selected from tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, oxazolyl, thiazolyl, imidazolyl, piperidinyl, piperazineyl, and morpholinyl.

[0068] In some implementations, R3 is selected from... Among them, the thick solid line This indicates that the attached carbon atom exhibits chiral isomerism.

[0069] In some implementations, R3 is selected from...

[0070] In some embodiments, the compound of formula (I) is a compound of formula (II):

[0071]

[0072] In the formula, R4, R5, R6, R7, and R9 are each defined as before.

[0073] In some specific implementations, R4 and R5 are hydrogen or fluorine; or R4 and R5 together with the carbon atoms they are attached to form cyclopropyl or cyclobutyl.

[0074] In some specific implementations, R6, R7, and R9 are each independently selected from hydroxyl, methyl, ethyl, and trifluoromethyl.

[0075] In some specific implementation plans, Selected from

[0076] In some embodiments, the compound of formula (I) is a compound of formula (III):

[0077]

[0078] In the formula, R4, R5, and X1 are each defined as before.

[0079] R A1 Selected from halogen, hydroxyl, oxo, cyano, nitro, C 1-6 Alkyl, C 1-6 Alkoxy group, -NHC(O)-C 1-6 Alkyl group, -NHS(O)2-C 1-6 Alkyl, C 3-10 Cycloalkyl and 3 to 10-membered heterocycloalkyl, wherein C 1-6 Alkyl, C 1-6 Alkoxy group, -NHC(O)-C 1-6 Alkyl group, -NHS(O)2-C 1-6 Alkyl, C 3-10 The cycloalkyl or 3- to 10-membered heterocycloalkyl group is optionally substituted by one or more substituents selected from halogens and hydroxyl groups;

[0080] m1 and n1 are each independently selected from 0, 1, 2 and 3.

[0081] In some specific implementation schemes, R4 and R5 are hydrogen.

[0082] In some specific implementations, X1 is N.

[0083] In some specific implementation schemes, R A1 It is an oxo group or a hydroxyl group, n1 is 0, 1 or 2, and n1 has 1 R. A1 Same or different.

[0084] In some specific implementations, m1 is 1.

[0085] In some specific implementation plans, for

[0086] In some embodiments, the compound of formula (I) is a compound of formula (IV):

[0087]

[0088] In the formula, Y is NH or CH2;

[0089] R A2 Selected from halogen, hydroxyl, oxo, cyano, nitro, C 1-6 Alkyl, C 1-6 Alkoxy group, -NHC(O)-C 1-6 Alkyl group, -NHS(O)2-C 1-6 Alkyl, C 3-10 Cycloalkyl and 3 to 10-membered heterocycloalkyl, wherein C 1-6 Alkyl, C1-6 Alkoxy group, -NHC(O)-C 1-6 Alkyl group, -NHS(O)2-C 1-6 Alkyl, C 3-10 The cycloalkyl or 3- to 10-membered heterocycloalkyl group is optionally substituted by one or more substituents selected from halogens and hydroxyl groups;

[0090] m2 and n2 are each independently selected from 0, 1, 2 and 3;

[0091] In some specific implementation schemes, R A2 Selected from methoxy, -(CH2)2-OH and -NH-C(O)-CH2-OH, n2 is 0, 1 or 2, n2 R A2 Same or different.

[0092] In some specific implementations, m2 is 0 or 1.

[0093] In some specific implementation plans, Selected from

[0094] In some embodiments, the compound of formula (I) is a compound of formula (V):

[0095]

[0096] In the formula, X2 is the same as before;

[0097] R A3 R A4 Each is independently selected from hydrogen, halogen, hydroxyl, oxo, cyano, nitro, C 1-6 Alkyl, C 1-6 Alkoxy group, -NHC(O)-C 1-6 Alkyl group, -NHS(O)2-C 1-6 Alkyl, C 3-10 Cycloalkyl and 3 to 10-membered heterocycloalkyl, wherein C 1-6 Alkyl, C 1-6 Alkoxy group, -NHC(O)-C 1-6 Alkyl group, -NHS(O)2-C 1-6 Alkyl, C 3-10 The cycloalkyl or 3- to 10-membered heterocycloalkyl group is optionally replaced by one or more substituents selected from halogens and hydroxyl groups.

[0098] In some specific implementations, X2 is CH or N.

[0099] In some specific implementation schemes, R A3 Selected from hydrogen, fluorine, and methoxy groups.

[0100] In some specific implementation schemes, RA4 Selected from hydroxyl and -(CH2)2-OH.

[0101] In some specific implementation plans, Selected from

[0102] This disclosure provides the following compounds, their pharmaceutically acceptable salts, stereoisomers, solvates, or prodrugs:

[0103]

[0104]

[0105] In another aspect of this disclosure, a method for preparing the above-mentioned compound is provided, comprising:

[0106] The condensation reaction of the -2'(S)-amino- of 7,9-naphthene in the eczetidine structure with the corresponding carboxylic acid-substituted compound forms an amide bond, yielding the corresponding eczetidine amide derivative.

[0107] The carboxylic acid-substituted compound is selected from... Any one of them.

[0108] In some embodiments of this disclosure, the substituents of the eczetidine derivative contain NH-substituted cycloalkyl groups. In this case, NH can react with an ethyl hydroxyl donor compound to obtain an ethyl hydroxyl-substituted eczetidine derivative.

[0109] In some embodiments of this disclosure, the substituents of the eczetidine derivative contain NH2. In this case, NH2 can undergo a nucleophilic addition reaction or cyclization reaction with 2-(2,2-dimethyl-5-oxo-1,3-dioxolane-4-yl)acetaldehyde to form a pyrrole ring, or undergo a condensation reaction with the corresponding carboxylic acid-substituted compound to form an amide bond to obtain an eczetidine derivative containing two amide bonds.

[0110] In another aspect of this disclosure, an antibody-drug conjugate is provided, comprising a small molecule drug, a linker, and an antibody, wherein the effective loading of the small molecule drug is the aforementioned compound or a pharmaceutically acceptable salt thereof.

[0111] In another aspect of this disclosure, a pharmaceutical composition is provided comprising the above-described compound or a pharmaceutically acceptable salt, stereoisomer, or prodrug, or the above-described antibody-drug conjugate, and a pharmaceutically acceptable excipient.

[0112] In another aspect of this disclosure, the use of the above-described compound or its pharmaceutically acceptable salt, stereoisomer or prodrug, the antibody-drug conjugate or the pharmaceutical composition thereof in the preparation of an antitumor drug is provided.

[0113] Another aspect of this disclosure provides a method for inhibiting tumors, comprising the step of administering a therapeutically effective amount of the aforementioned compound or a pharmaceutically acceptable salt, stereoisomer, or prodrug, antibody-drug conjugate, or pharmaceutical composition thereof to a patient in need.

[0114] The aforementioned compounds, or their pharmaceutically acceptable salts, stereoisomers, or prodrugs, may be used alone, in combination, or prepared as antibody-drug conjugates for the treatment of cancer.

[0115] In some embodiments of this disclosure, the tumor is selected from one or more of the following: liver cancer, lung cancer, colorectal cancer, pancreatic cancer, bone cancer, breast cancer, kidney cancer, esophageal cancer, ovarian cancer, oral cancer, laryngeal cancer, bile duct cancer, uterine cancer, nasal cancer, testicular cancer, meningioma, skin cancer, cervical cancer, glioma, lymphoma, leukemia, or sarcoma.

[0116] In some embodiments of this disclosure, the tumor is selected from one or more of lung cancer, breast cancer, colorectal cancer, and gastric cancer.

[0117] In some embodiments of this disclosure, the tumor and breast cancer are selected. Detailed Implementation

[0118] In this invention, unless otherwise stated, the scientific and technical terms used in this disclosure have the meanings commonly understood by those skilled in the art. Furthermore, the related terms and laboratory procedures used in this disclosure are all widely used terms and routine procedures in the respective fields. Meanwhile, to better understand this invention, definitions and explanations of related terms are provided below.

[0119] I. Definition

[0120] As used in this disclosure and unless otherwise stated, the terms “comprising,” “including,” “having,” “containing,” and their grammatically equivalent forms, including their grammatical equivalents, should generally be understood as open-ended and non-restrictive, e.g., not excluding other unlisted elements or steps.

[0121] This disclosure refers to compounds of formula (I), and also includes their tautomers, stereoisomers, mixtures of stereoisomers, solvates or derivatives.

[0122] In the description of this disclosure, references to “some embodiments,” “some implementations,” or “some implementation schemes” describe a subset of all possible embodiments. However, it is understood that “some embodiments” may be the same subset or different subsets of all possible embodiments and may be combined with each other without conflict.

[0123] As used in this disclosure and unless otherwise stated, the terms “comprising,” “including,” “having,” “containing,” and their grammatically equivalent forms, including their grammatical equivalents, should generally be understood as open-ended and non-restrictive, e.g., not excluding other unlisted elements or steps.

[0124] When listing numerical ranges, each value and subranges within that range are included by default. For example, “C1-6 alkyl” includes C1, C2, C3, C4, C5, C6, C1-6, C1-5, C1-4, C1-3, C1-2, C2-6, C2-5, C2-4, C2-3, C3-6, C3-5, C3-4, C4-6, C4-5, and C5-6 alkyl.

[0125] As used in this disclosure, the term "substitution" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include deuterium and hydrogen variants, provided that the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is an oxo group (i.e., =O), it means that two hydrogen atoms are replaced. The terms "optional substitution" or "optionally substituted" mean that substitution is optional, and unless otherwise specified, the type and number of substituents can be arbitrary on a chemically feasible basis.

[0126] As used in this disclosure, "pharmaceutically acceptable salt" refers to a salt formed by a corresponding amine compound and an inorganic or organic acid, or a salt formed by a corresponding carboxylic acid compound and an alkali or alkaline earth metal, or a salt formed by an organic amine. Inorganic acids include, but are not limited to, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, and phosphoric acid; organic acids include, but are not limited to, acetic acid, propionic acid, butyric acid, benzoic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, oxalic acid, succinic acid, lactic acid, citric acid, succinic acid, gluconic acid, maleic acid, fumaric acid, and tartaric acid; alkali or alkaline earth metal salts include, but are not limited to, sodium, potassium, calcium, and magnesium salts; organic amine salts include, but are not limited to, salts composed of ammonia, methylamine, ethylamine, propylamine, isopropylamine, dimethylamine, diethylamine, trimethylamine, triethylamine, tert-butylamine, ethylenediamine, ethanolamine, diethanolamine, triethanolamine, morpholine, piperidine, piperazine, and amino acids. In addition to salt forms, the compounds provided by this invention also exist as stereoisomers, solvates, and prodrug forms. The prodrugs of the compounds described in this disclosure readily undergo chemical changes under physiological conditions to be converted into the compounds of the present invention. Furthermore, the prodrugs can be converted into the compounds of the present invention in the in vivo environment via chemical or biochemical methods.

[0127] The term "stereoisomer" refers to an isomer formed due to at least one asymmetric center. In compounds having one or more (e.g., one, two, three, or four) asymmetric centers, racemic mixtures, single enantiomers, diastereomer mixtures, and individual diastereomers can be produced. Specific individual molecules may also exist as geometric isomers (cis / trans).

[0128] This disclosure of "compounds" also includes tautomer forms. A tautomer form arises from the exchange of a single bond with an adjacent double bond, accompanied by the migration of a proton. The terms "tautomer" or "tautomer form" refer to isomers of different functional groups in dynamic equilibrium at room temperature that can rapidly interconvert. It refers to one of two or more structural isomers that exist in equilibrium and readily transform from one isomer form to another. This transformation results in the formal migration of a hydrogen atom, accompanied by the conversion of adjacent conjugated double bonds. Tautomers exist as a mixture of tautomer groups in solution. In solutions where tautomerization is possible, chemical equilibrium of the tautomers will be reached. The exact proportions of the tautomers depend on several factors, including temperature, solvent, and pH conditions. The concept of tautomers that can interconvert through tautomerization is called tautomerism.

[0129] When this specification describes a compound that is readily tautomerizable, but only one of its tautomers is described, it should be understood that all tautomers are included as part of the chemical meaning described. It should be understood that when a compound has tautomeric forms, it is intended to include all tautomeric forms, and the naming of the compound does not exclude any tautomeric form.

[0130] Of the various possible types of tautomerism, two are typically observed. In keto-enol tautomerism, both electrons and hydrogen atoms move simultaneously.

[0131] Common tautomer pairs are: keto-enol, amide-nitrile, lactam-lactam, amide-imine tautomer in heterocycles, imine-enamine, and enamine-enamine.

[0132] The term "isomer" refers to different compounds having the same molecular formula but different atomic arrangements and configurations. Depending on their structure, the compounds of this disclosure can exist in different stereoisomeric forms. These forms include configurational isomers or optical conformational isomers (enantiomers and / or diastereomers, including those that are blocked from rotation). Therefore, this disclosure includes enantiomers, diastereomers, and mixtures thereof. This disclosure further includes all mixtures of the above-described stereoisomers, regardless of proportions, including racemic mixtures.

[0133] Depending on their structure, the compounds disclosed herein can exist in various stable isotopic forms. These forms include those in which one or more hydrogen atoms are replaced by deuterium atoms, those in which one or more nitrogen atoms are replaced by 15N atoms, or those in which one or more carbon, fluorine, chlorine, bromine, sulfur, or oxygen are replaced by stable isotopes of their respective original atoms.

[0134] According to this disclosure, some compounds and salts can exist in different crystalline forms (polymorphs) within the scope of this disclosure.

[0135] In this disclosure, the term is used. The absolute configuration representing the center of a solid. In It refers to the junction of chemical bonds.

[0136] When the ring appears Furthermore, if the connection location is uncertain, it indicates that the connection site is located at... Any atom on the monocyclic ring, as long as its valence allows.

[0137] As used in this disclosure, a "prodrug" refers to a compound that, after being administered to the human body via an appropriate route of administration, undergoes metabolism or simple chemical changes within the patient's body to transform into the compound and its corresponding salt contained in general formula (I) of this invention. Precursors to the compound include, but are not limited to, various carboxylic acid esters, carbonates, phosphate esters, sulfate esters, sulfonates, amino acid esters, gluconates, and various amides, acetals, hemiacetals, carbonate esters, etc.

[0138] The range of numbers in this disclosure refers to the integers within a given range. For example, "C 1-6 "" means that the group can have 1, 2, 3, 4, 5, or 6 carbon atoms; "C 3-6 "" means that the group can have 3, 4, 5 or 6 carbon atoms.

[0139] The term "cycloalkyl" refers to a saturated monocyclic or polycyclic (e.g., bicyclic) hydrocarbon ring (e.g., monocyclic, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or bicyclic, including spirocyclic, fused or bridged systems, such as bicyclic [2.2.1]heptyl, or polycyclic, including adamantyl, etc.). As used in this disclosure, the term "C" refers to a saturated monocyclic or polycyclic hydrocarbon ring. 3-10 "Cycloalkyl" refers to a saturated monocyclic or polycyclic (e.g., bicyclic) hydrocarbon ring (e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl) having 3 to 10 cyclic carbon atoms. Cycloalkyl groups may optionally be substituted with one or more suitable substituents.

[0140] The term "heterocyclic alkyl" refers to a saturated monocyclic or polycyclic (e.g., bicyclic) group having 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms and one or more heteroatoms in the ring; the heterocyclic alkyl group may be attached to the remainder of the molecule by any one of the carbon atoms or the heteroatom. As used in this disclosure, the term 3- to 10-membered heterocyclic alkyl is a saturated monocyclic or polycyclic (e.g., bicyclic) group having 3 to 10 cyclic atoms in the ring and containing at least one heteroatom that may be the same or different (the heteroatom is, for example, oxygen, nitrogen, or sulfur). Heterocyclic alkyl groups may optionally be substituted with one or more suitable substituents.

[0141] When any variable (e.g., R) A When a group appears more than once in the composition or structure of a compound, its definition is independent in each case. Therefore, for example, if a group is substituted by one or more R*, then the definition of R* in each case is independent. A Each has its own independent options. Furthermore, combinations of substituents and / or their variants are only permitted if such combinations produce stable compounds.

[0142] The term "halogen" is defined as including F, Cl, Br, or I.

[0143] The term "alkyl" is defined as a straight-chain or branched saturated aliphatic hydrocarbon group. As used in this disclosure, the term "C" is... 1-6 "Alkyl" refers to a straight-chain or branched saturated aliphatic hydrocarbon group having 1, 2, 3, 4, 5, or 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, etc. Alkyl groups may optionally be substituted with one or more suitable substituents.

[0144] The term "alkoxy" refers to an alkyl group connected by an oxygen atom to an alkyl group as defined above; that is, an "alkoxy" group can be defined as -OR, where R is an alkyl group as defined above. As used in this disclosure, the term "C"... 1-6 Examples of "alkoxy" include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, and n-hexoxy.

[0145] Term "C" 1-6"alkyl-OH" refers to an alkyl group as defined above, wherein one or more hydrogen atoms are replaced by hydroxyl groups, for example...

[0146] The term "optional substitution" refers to the optional substitution by a specific group, radical, or part thereof.

[0147] When a group is described as “optionally substituted with one or more substituents,” the group may be (1) unsubstituted or (2) substituted. If a carbon atom in a group is described as being optionally substituted with one or more substituents, one or more hydrogen atoms on the carbon atom (to the extent that any hydrogen atoms are present) may be substituted individually and / or together with independently selected substituents or be unsubstituted. If a nitrogen atom in a group is described as being optionally substituted with one or more substituents, one or more hydrogen atoms on the nitrogen atom (to the extent that any hydrogen atoms are present) may each be substituted with independently selected substituents or be unsubstituted.

[0148] The compounds disclosed herein may also contain one or more (e.g., one, two, three or four) isotope substitutions.

[0149] All hydrogen atoms described in this invention can be replaced by their isotope deuterium.

[0150] "Substituted" refers to one or more hydrogen atoms in a group, preferably up to five, and more preferably one to three hydrogen atoms, which are independently substituted by the corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without much effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom having an unsaturated bond (such as an alkene).

[0151] Drugs or drug compositions

[0152] The pharmaceuticals or pharmaceutical compositions of the present invention can be administered orally, topically, parenterally, or mucosally (e.g., sublingually, by inhalation, or rectally) in dosage units comprising conventional, non-toxic, pharmaceutically acceptable carriers. Oral administration is generally preferred. The active agent can be administered orally in capsule, tablet, or other similar forms.

[0153] For oral administration in tablet or capsule form, the active pharmaceutical ingredient may be combined with non-toxic, pharmaceutically acceptable excipients such as binders (e.g., pregelatinized corn starch, polyvinylpyrrolidone, or hydroxypropyl methylcellulose); fillers (e.g., lactose, sucrose, glucose, mannitol, sorbitol, and other reducing and non-reducing sugars, microcrystalline cellulose, calcium sulfate, or calcium hydrogen phosphate); lubricants (e.g., magnesium stearate, talc or silica, stearic acid, sodium stearyl fumarate, glyceryl docosanoate, calcium stearate, etc.); disintegrants (e.g., potato starch or sodium hydroxyacetic acid starch); or wetting agents (e.g., sodium lauryl sulfate), colorants and flavorings, gelatin, sweeteners, natural and synthetic gums (e.g., gum arabic, tragacanth, or alginate), buffer salts, carboxymethyl cellulose, polyethylene glycol, waxes, etc. For oral administration in liquid form, the pharmaceutical component may be combined with a non-toxic, pharmaceutically acceptable inert carrier (e.g., ethanol, glycerol, water), an anti-settling agent (e.g., sorbitol syrup, cellulose derivatives, or hydrogenated edible fats), an emulsifier (e.g., lecithin or gum arabic), a non-aqueous carrier (e.g., almond oil, esters, ethanol, or fractionated vegetable oils), and a preservative (e.g., methylparaben, propylparaben, or sorbic acid). Stabilizers such as antioxidants (BHA, BHT, propyl iodide, sodium ascorbate, citric acid) may also be added to stabilize the dosage form.

[0154] Tablets containing the active compound can be coated using methods well known in the art. The compositions of the present invention containing a compound of formula I as the active compound can also incorporate beads, microspheres, or microcapsules, for example, constructed from polyglycolic acid / lactic acid (PGLA). Liquid formulations for oral administration can take the form of, for example, solutions, syrups, emulsions, or suspensions, or they can be presented as dry products reconstituted with water or other suitable excipients prior to use. Formulations for oral administration can be suitably formulated to allow for controlled or delayed release of the active compound.

[0155] The term "pharmaceutical composition" means a composition comprising the compound described in this invention or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable ingredient selected from the following, depending on the manner of administration and dosage form: carrier, diluent, adjuvant, excipient, preservative, filler, disintegrant, wetting agent, emulsifier, suspending agent, sweetener, flavoring agent, fragrance, antibacterial agent, antifungal agent, lubricant, dispersant, thermosensitive material, temperature regulator, adhesive, stabilizer, suspending agent, etc.

[0156] The term "antibody-drug conjugate" (ADC) refers to a small molecule drug with biological activity linked to a monoclonal antibody via a chemical link. The monoclonal antibody then acts as a carrier to target and deliver the small molecule drug to target cells.

[0157] The term "treatment" includes suppressing, alleviating, preventing, or eliminating one or more symptoms or side effects associated with the disease, condition, or disorder being treated.

[0158] The term "inhibition" is used relative to a control. Those skilled in the art will readily determine the appropriate control for each experiment. For example, a reduced response in a subject or cell treated with the compound is compared to a response in a subject or cell not treated with the compound.

[0159] The term "effective dose" or "therapeutic effective dose" refers to a sufficient amount of a drug or agent that is non-toxic but achieves the desired effect. In embodiments of the invention, when treating a patient according to the invention, the amount of a given drug depends on many factors, such as the specific dosing regimen, the type and severity of the disease or condition, and the unique characteristics of the patient or host requiring treatment (e.g., weight). However, depending on the specific surrounding circumstances, including, for example, the specific drug used, the route of administration, the condition being treated, and the patient or host being treated, the dosage can be conventionally determined by methods known in the art. Typically, for adult treatment, the dosage is typically in the range of 0.02-5000 mg / day, for example, about 1-1500 mg / day. This required dose can conveniently be expressed as a single dose, or concurrent (or over a short period of time) or fractions at appropriate intervals, such as two, three, four, or more doses per day. Those skilled in the art will understand that although the above dosage ranges are given, the specific effective dose can be appropriately adjusted according to the patient's condition and in conjunction with the physician's diagnosis.

[0160] Unless otherwise specified, the raw materials and equipment used in the specific embodiments of the present invention are all known products and are obtained by purchasing commercially available products.

[0161] II. Specific Implementation Examples

[0162] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be described in further detail below. The described embodiments should not be considered as limitations on the invention. All other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0163] The structures of the compounds disclosed herein were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR chemical shifts (δ) are given in parts per million (ppm). NMR measurements were performed using an AVANCE NEO 400MHz Bruker instrument with deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD) as solvents, and tetramethylsilane (TMS) as an internal standard. MS measurements were performed using an ISQ-EC Thermo Fisher LC-MS instrument.

[0164] Example 1: Preparation of compounds 21 and 23

[0165]

[0166] Experimental Procedure: Ecitec mesylate (100 mg, 188.128 μmol, 1 eq, purchased from Anqing Runke Biomedical Technology Co., Ltd.) was dissolved in H2O (0.8 mL) and DMF (3.2 mL). Then, 6 (37 mg, 234.065 μmol, 1.244 eq, purchased from Yaoshi Technology Co., Ltd.), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholin-4-ium chloride (104 mg, 375.833 μmol, 1.998 eq), and DIPEA (72 mg, 557.094 μmol, 2.961 eq) were added. The mixture was then incubated at 25 °C for 1 h. TLC analysis showed the reaction was complete. 6 mL of water was then added. A solid precipitated. The sample was filtered, washed with 2 mL x 3 of water, and then prepared by reverse phase. This yielded pale yellow solids 21 (2 mg, 4% yield) and 23 (10 mg, 18% yield).

[0167] 1 H NMR (400MHz, DMSO-d6) δ8.63(d,J=8.6Hz,1H),7.81(d,J=11.0Hz,1H),7.32(s,1H),6.53-6.45(m,2H),5.59(dt,J=8.8,4.5Hz,1H),5.45-5.16(m ,4H),4.43(s,1H),3.21-3.17(m,2H),2.53-2.51(m,1H),2.48-2.36(m, 4H),2.21-2.12(m,2H),1.96-1.75(m,2H),0.85(dt,J=10.8,7.1Hz,3H).

[0168] LC-MS (m / z): 576.2 [M+H] +

[0169] 1 H NMR (400MHz, DMSO-d6) δ8.62(d,J=8.5Hz,1H),7.82(d,J=11.0Hz,1H),7.32(s,1H),6.43(s,2H),5.65-5.55(m,1H),5.50-5.26(m,4H ),4.42(s,1H),3.19-3.16(m,2H),2.49-2.40(m,5H),2.27-2.15(m,1H),2.15-2.03(m,1H),1.89-1.82(m,2H),0.88(t,J=7.3Hz,3H).

[0170] LC-MS (m / z): 576.2 [M+H] +

[0171] Example 2: Preparation of compounds 22-1 and 22-2

[0172]

[0173] Experimental Procedure: Eciticon mesylate (100 mg, 188.128 μmol, 1 eq, purchased from Anqing Runke Biomedical Technology Co., Ltd.) was dissolved in 0.8 mL H2O and 3.2 mL DMF. Then, 7 (40 mg, 232.420 μmol, 1.235 eq, purchased from Yaoshi Technology Co., Ltd.), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholin-4-ium chloride (104 mg, 375.833 μmol, 1.998 eq), and DIPEA (73 mg, 564.832 μmol, 3.002 eq) were added. The mixture was then incubated at 25 °C for 1 h. TLC analysis showed the reaction was complete. 6 mL of water was then added. A solid precipitated. Filter the solution, wash with 2 ml of water three times, and use the resulting solid for direct reverse-phase preparation. This yielded pale yellow solids 22-1 (4 mg, 7% yield) and 22-2 (17 mg, 30% yield).

[0174] 1H NMR(400MHz,DMSO-d6)δ8.70-8.47(m,1H),7.89-7.68(m,1H),7.32(s,1H),6.53(s,1H),6.11(s,1H),5.63-5.59(m,1H),5.46-5.13(m,4H ),3.19(s,2H),2.58-2.52(m,1H),2.47-2.35(m,4H),2.27-2.02(m,2H),1.99-1.75(m,2H),1.44(d,J=7.6Hz,3H),0.88(t,J=7.3Hz,3H).

[0175] LC-MS (m / z): 590.2 [M+H] +

[0176] 1 H NMR (400MHz, DMSO-d6) δ8.58(d,J=8.5Hz,1H),7.82(d,J=11.0Hz,1H),7.32( s,1H),6.53(s,1H),6.24(s,1H),5.57(dt,J=8.7,4.3Hz,1H),5.43(s,2H),5 .28-5.15(m,2H),3.18(d,J=4.8Hz,2H),2.59-2.51(m,1H),2.47-2.29(m,4H ),2.21-2.10(m,2H),1.96-1.72(m,2H),1.43(s,3H),0.88(t,J=7.3Hz,3H).

[0177] LC-MS (m / z): 590.2 [M+H] +

[0178] Example 3: Preparation of compounds 33-1 and 33-2

[0179]

[0180] Experimental steps:

[0181] Eciticon mesylate (50 mg, 94.064 μmol, 1 eq, purchased from Anqing Runke Biomedical Technology Co., Ltd.) was dissolved in H2O (0.8 mL) and DMF (3.2 mL). Then, 3a (23 mg, 117.202 μmol, 1.246 eq, purchased from Yaoshi Technology Co., Ltd.), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholin-4-ium chloride (52 mg, 187.916 μmol, 1.998 eq) and DIPEA (36 mg, 278.547 μmol, 2.961 eq) were added, and the mixture was reacted at 25 °C for 1 h. TLC analysis showed the reaction was complete. Adding 6 mL of water resulted in the precipitation of a solid. Filter and rinse with 2 mL x 3 of water. Isomers were present; the mixture was resolved by reverse-phase chromatography to give 14 mg of a mixture of compounds 33-1 and 33-2. Overall yield: 24%.

[0182] 1 H NMR (400MHz, DMSO-d6) δ8.04-7.91(m,1H),7.77(d,J=11.0Hz,1H),7.30(s,1H),6.51(s,1H),5.61-5.51(m,1H),5.49-4.99(m,4H),4. 66-4.56(m,1H),3.71-3.56(m,1H),3.24-3.05(m,2H),2.39(s,3H),2.31-1.51(m,15H),1.33(d,J=11.7Hz,1H),0.88(t,J=7.3Hz,3H).

[0183] LC-MS (m / z): 614.3 [M+H] +

[0184] Example 4: Preparation of Compound 40

[0185]

[0186] Experimental steps:

[0187] Step 1: Dissolve eczemab mesylate (100 mg, 188.128 μmol, 1 eq, purchased from Anqing Runke Biomedical Technology Co., Ltd.) in DMF (3.2 mL) and H2O (0.8 mL). Add 1a (50 mg, 232.293 μmol, 1.235 eq, purchased from Yaoshi Technology Co., Ltd.), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholin-4-ium chloride (104 mg, 375.833 μmol, 1.998 eq), and DIPEA (72 mg, 557.094 μmol, 2.961 eq). Then, react at 25°C for 3 hours. TLC detection showed the reaction was complete. Adding 6 mL of water resulted in solid precipitation. Filter and wash with 10 mL of water. A pale yellow solid, 1b, 106 mg, yield 89%.

[0188] 1 H NMR(400MHz,DMSO-d6)δ8.38(d,J=8.7Hz,1H),7.79(d,J=11.0Hz,1H),7.30(s, 1H),7.17(d,J=8.2Hz,1H),6.52(s,1H),5.64-5.55(m,1H),5.49-5.05(m,4H),4 .30-4.11(m,1H),3.27-3.10(m,2H),2.94-2.83(m,1H),2.47-2.27(m,5H),2.20 -2.04(m,4H),1.86(qt,J=14.2,7.3Hz,2H),1.38(s,9H),0.88(t,J=7.3Hz,3H).

[0189] LC-MS (m / z): 633.42 [M+H] +

[0190] Step 2: Dissolve 1b (106 mg, 167.542 μmol, 1 eq) in DCM (2 mL), then add TFA (0.4 mL) and react at 25 °C for 16 h. The reaction was complete as determined by TLC. The reaction solution was directly evaporated to dryness. Without purification, proceed directly to the next step. A yellow solid 1c, 89 mg, was obtained; yield was calculated as 100%. LC-MS (m / z): 533.5 [M+H] +

[0191] Step 3: 1 c (89 mg, 167.117 μmol, 1 eq) was placed in 0.8 mL H2O and 3.2 mL DMF. Glycolic acid (15 mg, 197.236 μmol, 1.180 eq), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholin-4-ium chloride (69 mg, 249.350 μmol, 1.492 eq), and DIPEA (64 mg, 495.195 μmol, 2.963 eq) were added. The mixture was then incubated at 25 °C for 1 h. TLC analysis confirmed the reaction was complete. The reaction solution was then directly reversed to prepare the product. 40.5 mg of a pale yellow solid was obtained, with a yield of 5%.

[0192] 1 H NMR (400MHz, DMSO-d6) δ8.41(d,J=8.6Hz,1H),8.00(d,J=8.2Hz,1H),7.75(d,J=10.9 Hz,1H),7.29(s,1H),6.52(s,1H),5.59(dd,J=2.9,1.3Hz,1H),5.49-5.32(m,3H),5. 24-5.03(m,2H),4.64-4.50(m,1H),3.78(s,2H),3.25-3.06(m,2H),2.93(t,J=8.9Hz ,1H),2.44-2.23(m,7H),2.23-2.04(m,2H),1.94-1.76(m,2H),0.87(t,J=7.1Hz,3H).

[0193] LC-MS (m / z): 591.3 [M+H] +

[0194] Example 5: Preparation of Compound 41

[0195]

[0196] Experimental steps:

[0197] Step 1: Dissolve eczemacin mesylate (100 mg, 188.128 μmol, 1 eq, purchased from Anqing Runke Biomedical Technology Co., Ltd.) in H2O (0.8 mL) and DMF (3.2 mL). Add 2a (47 mg, 233.576 μmol, 1.242 eq, purchased from Yaoshi Technology Co., Ltd.), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholin-4-ium chloride (104 mg, 375.833 μmol, 1.998 eq), and DIPEA (73 mg, 564.832 μmol, 3.002 eq) and react at 25 °C for 3 h. After the reaction is complete, add 6 mL of water; a solid precipitates out. Filter and wash with 10 mL of water. A pale yellow solid, 2b, 114 mg, was obtained, with a yield of 97%.

[0198] 1 H NMR (400MHz, DMSO-d6) δ8.58(d,J=8.7Hz,1H),7.81(d,J=11.0Hz,1H),7.31(s,1H),6.52(s,1H),5.74-5.52(m,1H),5.54-4.97(m,4H),3.94 (s,4H),3.28(d,J=7.2Hz,1H),3.17(t,J=6.0Hz,2H),2.41(s,3H),2.21-2.06(m,2H),1.88-1.86(m,2H),1.39(s,9H),0.88(t,J=7.3Hz,3H).

[0199] LC-MS (m / z): 619.4 [M+H] + Step 2: Dissolve 2b (114 mg, 184.272 μmol, 1 eq) in DCM (2 mL), add TFA (0.4 mL), and then react at 25 °C for 16 h. The reaction was complete as determined by TLC. The reaction solution was directly evaporated to dryness. Without purification, proceed directly to the next step. A yellow solid 2c, 95 mg, was obtained; yield was calculated as 100%. LC-MS (m / z): 519.4 [M+H] +

[0200] Step 3: Dissolve 2c (95 mg, 183.208 μmol, 1 eq) in MeOH (2 mL), add sodium cyanoborohydride (57 mg, 907.037 μmol, 4.951 eq) and 1,4-Dioxane-2,5-diol (110 mg, 915.873 μmol, 4.999 eq), displace nitrogen gas, and react at 25 °C for 16 h. TLC detection showed the reaction was complete. The reaction solution was then directly reversed to prepare the product. 41,30 mg of a yellow solid was obtained, with a yield of 29%.

[0201] 1 H NMR (400MHz, DMSO-d6) δ10.14-9.80(m,1H),8.70(d,J=8.4Hz,1H),7.81(d,J=10.9Hz,1H),7.32(s,1H),6.54(s,1H),5.74-5.52(m,1H),5.50-5 .10(m,4H),4.34-4.05(m,4H),3.66-3.56(m,3H),3.32-3.08(m,4H),2. 41(s,3H),2.29-2.04(m,2H),1.96-1.74(m,2H),0.87(t,J=7.3Hz,3H).

[0202] LC-MS (m / z): 563.3 [M+H] +

[0203] Example 6: Preparation of Compound 45

[0204]

[0205] Experimental steps:

[0206] Step 1: Dissolve 5a (10.00g, 74.578mmol, 1eq, purchased from Pharmaron Technology Co., Ltd.) in 2,2-dimethoxypropane (250mL), and slowly add p-toluenesulfonic acid (642mg, 3.728mmol, 4.999e) to the solution. -2 The solution was then incubated at 25°C for 17 hours. TLC was performed to confirm completion. The reaction solution was directly evaporated to dryness and then purified to form granules. Column chromatography (PE:EA = 1:1) was used to obtain 3.00 g of the target product 5b, with a yield of 23%.

[0207] 1H NMR (400MHz, CDCl3) δ10.18(s,1H),4.73(dt,J=6.5,4.1Hz,1H),3.75(s,1H),2.97-2.91(m,1H),2.90-2.81(m,1H),1.62(s,3H),1.56(s,3H).

[0208] Step 2: Dissolve 5b (1.00 g, 5.742 mmol, 1 eq) in DCM (25 mL), then cool to 0 degrees Celsius and add ethyl mercaptan (321 mg, 5.166 mmol, 8.997 e eq). -1 eq), DCC (1.54 g, 7.464 mmol, 1.30 eq) and DMAP (14 mg, 114.597 μmol, 1.996 e eq) -2 The mixture was heated to room temperature and allowed to react for 17 hours. The reaction was confirmed complete by TLC. The reaction solution was filtered, and the filtrate was washed with 10 mL of saturated brine. The resulting organic phase was dried and then evaporated to dryness. The resulting precipitate was prepared by column chromatography using a PE:EA ratio of 10:1. A yellow oily substance, 5c, 500 mg, was obtained, with a yield of 39%.

[0209] 1 H NMR (400MHz, CDCl3) δ4.82-4.72(m,1H),3.19-3.10(m,1H),3.09-2.88(m,3H),1.64(s,3H),1.58(s,3H),1.29(t,J=7.0Hz,5H),1.00-0.90(m,1H).

[0210] Step 3: Dissolve 5c (200 mg, 916.298 μmol, 1 eq) in acetone (4 mL), then place the reaction solution in an ice bath and cool to 0 degrees Celsius. Next, add palladium acetate (41 mg, 182.622 μmol, 1.993 e) to the solution. -1 Triethylsilane (213 mg, 1.833 mmol, 2 eq) was reacted at room temperature for 17 h. TLC analysis showed the formation of new spots. The reaction mixture was filtered and washed with EA. The solution was then prepared and subjected to PE:EA 1:1 column chromatography. A brown oily substance was obtained, 100 mg, for 5 days, in 69% yield.

[0211] 1H NMR (400MHz, CDCl3) δ9.80 (s, 1H), 4.81 (dd, J = 3.5, 6.8Hz, 1H), 3.15-2.92 (m, 2H), 1.65 (s, 3H), 1.60 (s, 3H)

[0212] Step 4: Dissolve eczemab mesylate (100 mg, 188.128 μmol, 1 eq, purchased from Anqing Runke Biomedical Technology Co., Ltd.) in H2O (0.8 mL) and DMF (3.2 mL). Add BOC-glycine (40 mg, 228.334 μmol, 1.214 eq), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholin chloride (104 mg, 375.833 μmol, 1.998 eq), and DIPEA (73 mg, 564.385 μmol, 3 eq) and react at 25 °C for 3 h. The reaction was confirmed complete by TLC. Add 6 mL of water; a solid precipitated. Filter and wash with 10 mL of water. A pale yellow solid, 5e, 111 mg, was obtained, with a yield of 100%.

[0213] 1 H NMR (400MHz, DMSO-d6) δ8.39(d,J=8.6Hz,1H),7.80(d,J=11.0Hz,1H),7.31(s,1H),6.93(t,J=5.8Hz,1H),6.55(s,1H),5.66-5.49(m,1H), 5.49-5.14(m,4H),3.65-3.55(m,2H),3.17(s,2H),2.40(s,3H),2.25-2.01(m,2H),1.90-1.81(m,2H),1.26(s,9H),0.86(t,J=7.3Hz,3H).

[0214] Step 5: Dissolve 5e (111 mg, 187.306 μmol, 1 eq) in DCM (2 mL), add TFA (0.4 mL), and then react at 20 °C for 16 h. TLC analysis showed the reaction was complete. Directly evaporate to dryness and proceed to the next step. A yellow solid 5f, 92 mg, was obtained; yield was calculated as 100%.

[0215] LC-MS (m / z): 493.5 [M+H] +

[0216] Step Six: Dissolve 5f (92 mg, 187.554 μmol, 1 eq) in DMA (1 mL) and DCE (4 mL), then add 5d (30 mg, 189.692 μmol, 1.011 eq), followed by acetic acid (17 mg, 283.088 μmol, 1.509 eq). Stir at 25°C for 30 min, then add sodium triacetoxyborohydride (61 mg, 287.817 μmol, 1.535 eq), and stir at 25°C for 16 hours. The reaction was confirmed complete by TLC. Add 30 mL of saturated brine and extract using DCM:MeOH = 5:1 (20 mL * 3). Combine the organic phases and dry. Then reverse the phase to prepare the final product. 45.5 mg of a yellow solid was obtained, yield 4.6%.

[0217] 1 H NMR (400MHz, DMSO-d6) δ8.64(dd,J=8.4,3.9Hz,1H),7.81(d,J=10.9Hz,1H),7.32(s,1H),6.54(s,1H),5.62-5.48(m,2H),5.43-5.22( m,4H),4.14(s,1H),4.08-3.68(m,2H),3.19(s,4H),2.41(s,3H),2.33(s,1H),2.17(s,2H),2.00-1.80(m,3H),0.88(t,J=7.3Hz,3H).

[0218] LC-MS (m / z): 577.2 [M+H] +

[0219] Example 7: Preparation of Compound 49-1 and Compound 49-2

[0220]

[0221] Experimental steps:

[0222] Step 1: Dissolve 9a (2.00g, 14.069mmol, 1eq, purchased from Pharmaron Technology Co., Ltd.) in THF (30mL), and add cesium fluoride (213mg, 1.402mmol, 9.966e) to it. -2The mixture was first heated to 0°C, and then (trifluoromethyl)trimethylsilane (4.00 g, 28.130 mmol, 1.999 eq) was added. The temperature was then raised to 15°C and the reaction was allowed to proceed for 6 hours. TLC analysis showed no residue of the starting material. 7.7 mL of 4N HCl was added, and the reaction was allowed to continue for 16 hours. TLC analysis confirmed the reaction was complete. A saturated sodium bicarbonate solution was added to the reaction mixture, and extraction was performed using EA. The mixture was dried and prepared as a granulation sample. Column chromatography using PE:EA = 5:1 yielded 1.50 g of the target product 9b, with a yield of 50%.

[0223] 1 H NMR (400MHz, CD3OD) δ4.77 (q, J=6.6Hz, 1H), 4.30-3.96 (m, 2H), 1.39 (ddd, J=1 0.0,7.1,4.1Hz,1H),1.30-1.20(m,4H),1.17-1.04(m,1H),0.99-0.87(m,1H).

[0224] Step 2: Dissolve 9b (1.00 g, 4.713 mmol, 1 eq) in THF (10 mL), water (3 mL), and MeOH (10 mL). Cool to 0°C, then add LiOH·H₂O (593 mg, 14.131 mmol, 2.998 eq). Naturally heat to 15°C and react for 16 h. TLC detection indicates the reaction is complete. Discard the organic phase from the reaction solution, extract with EA, adjust the pH of the aqueous phase to 2, and then extract with MTBE. Dry the obtained organic phase to give 9c, 867 mg, which was directly used in the next step without purification. Yield is calculated as 100%.

[0225] 1 H NMR (400MHz, DMSO-d6) δ12.39 (s, 1H), 6.44 (d, J = 6.2Hz, 1H), 4.84-4.59 (m, 1H), 1.30-1.21 (m, 1H), 1.19-0.82 (m, 3H).

[0226] Step 3: Iciticon mesylate (191 mg, 359.325 μmol, 1.103 eq, purchased from Anqing Runke Biomedical Technology Co., Ltd.) was dissolved in DMF (2 mL). 9c (60 mg, 325.887 μmol, 1.0 eq), DIPEA (139 mg, 1.076 mmol, 3.300 eq), and 1H-benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (281 mg, 539.978 μmol, 1.657 eq) were added, and the mixture was reacted at 25 °C for 1 h. TLC analysis confirmed the reaction was complete. 6 mL of water was added. The precipitated solid was directly reverse-phase to prepare diastereomers 49-1 and 49-2, 5 mg each. Yield: 5%.

[0227] 1 H NMR(400MHz,DMSO-d6)δ8.26(d,J=8.4Hz,1H),7.80(d,J=11.0Hz,1H),7.31(s,1H) ,6.70(d,J=6.7Hz,1H),6.53(s,1H),5.60(d,J=7.8Hz,1H),5.43(s,2H),5.23-4.9 8(m,2H),4.77-4.41(m,1H),3.19-3.13(m,2H),2.40(s,3H),2.22-2.01(m,2H),1. 95-1.77(m,2H),1.26(dd,J=10.9,7.1Hz,1H),0.99(s,1H),0.88(t,J=7.3Hz,5H).

[0228] LC-MS (m / z): 602.2 [M+H] +

[0229] 1 H NMR (400MHz, DMSO-d6) δ8.17(d,J=8.0Hz,1H),7.79(d,J=10.4Hz,1H),7.31(s,1H),7.01(d,J=6.1Hz,1H),6.53(s,1H),5.61(s,1H),5.43(s,2H), 5.10-5.07(m,2H),4.34(s,1H),3.16(s,2H),2.40(s,3H),2.11(s,2H),1 .87(s,2H),1.24(s,2H),1.13(s,1H),0.99(s,1H),0.87(d,J=6.6Hz,3H).

[0230] LC-MS (m / z): 602.2 [M+H] +

[0231] Example 8: Preparation of Compound 50

[0232]

[0233] Experimental steps:

[0234] Step 1: Dissolve 8a (220 mg, 1.409 mmol, 1 eq, purchased from Shanghai Haohong Biomedical Technology Co., Ltd.) in THF (5 mL), and add cesium fluoride (21 mg, 138.246 μmol, 9.814 e) to it. -2 The mixture was first reacted with trifluoromethyl trimethylsilane (400 mg, 2.813 mmol, 1.997 eq) and then cooled to 0°C. The mixture was then heated to 15°C and reacted for 6 hours. TLC analysis showed no residue of the starting material. 0.77 mL of 4N HCl was added, and the reaction was continued for 16 hours. TLC analysis confirmed the reaction was complete. A saturated sodium bicarbonate solution was added to the reaction mixture, and extraction was performed using EA. The mixture was dried and prepared as a granulation sample. Column chromatography using PE:EA = 5:1 yielded 192 mg of the target product 8b, with a yield of 60%.

[0235] 1 H NMR (400MHz, CDCl3) δ4.29(q,J=7.1Hz,2H),4.24-4.09(m,1H),3.95(t,J=10.0Hz,1H),2.68-2.57(m,1 H),2.57-2.47(m,1H),2.41-2.29(m,1H),2.29-2.17(m,1H),2.15-1.86(m,2H),1.34(t,J=7.1Hz,3H).

[0236] Step 2: Dissolve 8b (192 mg, 848.835 μmol, 1 eq) in THF (2 mL), water (0.6 mL), and MeOH (2 mL). Cool to 0°C, then add LiOH·H₂O (107 mg, 2.550 mmol, 3.004 eq). Naturally heat to 15°C and react for 16 h. TLC detection confirms completion. Remove the organic phase from the reaction solution by rotary evaporation, extract with EA, adjust the pH to 2 in a water bath, and then extract with MTBE. Dry the obtained organic phase to give 8c, 168 mg, which is directly used in the next step without purification. Yield is calculated as 100%.

[0237] 1H NMR(400MHz,DMSO-d6)δ12.32(s,1H),6.77(d,J=7.0Hz,1H),4.29-4.08(m,1H),2.62-2.53(m,1H) ,2.26(t,J=8.1Hz,2H),1.97(ddd,J=11.4,6.5,3.0Hz,1H),1.90-1.81(m,1H),1.78-1.64(m,1H).

[0238] Step 3: Iciticon mesylate (180 mg, 338.631 μmol, 1.118 eq, purchased from Anqing Runke Biomedical Technology Co., Ltd.) was dissolved in DMF (2 mL). 8c (60 mg, 302.817 μmol, 1.0 eq), DIPEA (131 mg, 1.014 mmol, 3.347 eq), and 1H-benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (264 mg, 507.311 μmol, 1.675 eq) were added, and the mixture was reacted at 25 °C for 1 h. TLC analysis confirmed the reaction was complete. 6 mL of water was added, resulting in solid precipitation. The product was then directly prepared by reverse phase reaction. 50 μL of yellow solid was obtained, totaling 34 mg, with an overall yield of 18%.

[0239] 1 H NMR (400MHz, DMSO-d6) δ8.30-8.12(m,1H),7.80(d,J=11.0Hz,1H),7.31(s,1H),6.94-6.58(m,1H),6.53(s,1H),5.70-5.5 2(m,1H),5.42(s,2H),5.16-5.05(m,2H),4.30-4.27(m,1H),3.25-2.98(m,2H),2.47-1.59(m,13H),0.88(t,J=7.3Hz,3H).

[0240] LC-MS (m / z): 616.2 [M+H] +

[0241] Example 9: Preparation of compounds 52-1 and 52-2

[0242]

[0243] Experimental steps:

[0244] Step 1: Dissolve 2-amino-2-(bicyclo[1.1.1]pentan-1-yl)acetic acid hydrochloride (1.00 g, 5.630 mmol, 1 eq, purchased from Pharmaron) in 1M sulfuric acid (1.10 g, 11.216 mmol, 1.992 eq), cool to 0°C, and add sodium nitrite (2.33 g, 33.771 mmol, 5.999 eq) dropwise over half an hour. Then, allow the mixture to naturally warm to 15°C and react for 16 hours. TLC analysis showed no residual starting material. Add NaCl to the solution, then extract with MTBE and dry the organic phase. Reductize to dryness to obtain the unpurified product directly for the next step. Yield is 800 mg (100% yield).

[0245] LC-MS (m / z): 141.0 [MH] -

[0246] Step 2: Iciticon mesylate (100 mg, 188.128 μmol, 1 eq, purchased from Anqing Runke Biomedical Technology Co., Ltd.) was dissolved in DMF (2 mL). 4b (32 mg, 225.111 μmol, 1.197 eq), DIPEA (73 mg, 564.832 μmol, 3.002 eq), and 1H-benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (147 mg, 282.480 μmol, 1.502 eq) were added, and the mixture was reacted at 25 °C for 1 h. TLC analysis showed the reaction was complete. 6 mL of water was added. A solid precipitated. The mixture was filtered, washed with 2 mL of water three times, and then directly prepared by reverse phase. 52-1 (5 mg) and 52-2 (7 mg) were obtained, respectively.

[0247] 1H NMR (400MHz, DMSO-d6) δ8.29(d,J=8.7Hz,1H),7.78(d,J=10.9Hz,1H),7.31(s,1H),6.54(s,1H),5.59-5.50(m,1H),5.48-5.07 (m,4H),3.89(s,1H),3.24-3.03(m,3H),2.47(s,1H),2.39(s,3H),2.23-2.03(m,2H),1.93-1.66(m,8H),0.87(t,J=7.3Hz,3H).

[0248] LC-MS (m / z): 560.2 [M+H] +

[0249] 1 H NMR (400MHz, DMSO-d6) δ8.22(d,J=8.8Hz,1H),7.77(d,J=10.9Hz,1H),7.31(s,1H),6.54(s,1H),5.58-5.48(m,2H),5.46-5.05(m,4H),3.94( d,J=4.9Hz,1H),3.23-3.06(m,2H),2.46(s,1H),2.39(s,3H),2.26-2. 05(m,2H),1.95-1.77(m,2H),1.77-1.63(m,6H),0.87(t,J=7.3Hz,3H).

[0250] LC-MS (m / z): 560.2 [M+H] +

[0251] Example 10: Preparation of Compounds 53 and 54

[0252]

[0253] Experimental Procedure: Eciticon mesylate (100 mg, 188.128 μmol, 1 eq, purchased from Anqing Runke Biomedical Technology Co., Ltd.) was dissolved in H2O (0.8 mL) and DMF (3.2 mL). 10 (34 mg, 220.619 μmol, 1.173 eq, Shanghai Haohong Biomedical Technology Co., Ltd.) of 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholin-4-ium chloride ((4,6-Dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholin-4-ium chloride) (104 mg, 375.833 μmol, 1.998 eq) and DIPEA (72 mg, 557.094 μmol, 2.961 eq) were added. The mixture was then placed at 25 °C and reacted for 2 h. TLC analysis showed the reaction was complete. 6 mL of water was added, and a solid precipitated. The mixture was then filtered. The mixture was prepared by reverse phase separation, yielding 53 and 54, 5 mg each, for a total yield of 9%.

[0254] 1 H NMR (400MHz, DMSO-d6) δ9.33(d,J=8.3Hz,1H),7.81(d,J=11.0Hz,1H),7.32(s,1H),6.53(s,1H),5.73(s,1H),5.59(d,J=6.6Hz,1H),5.43(s ,2H),5.29-5.09(m,2H),3.84(s,1H),3.17(s,2H),2.41(s,3H),2.30- 2.08(m,2H),1.90-1.83(m,2H),1.59-1.33(m,2H),0.97-0.86(m,6H).

[0255] LC-MS (m / z): 572.2 [M+H] +

[0256] 1 H NMR (400MHz, DMSO-d6) δ9.31(d,J=8.4Hz,1H),7.80(d,J=11.0Hz,1H),7.32(s,1H),6.54(s,1H),5.73-5.54(m,1H),5.43(s,2H),5.29-5 .10(m,2H),3.87(d,J=6.4Hz,1H),3.18(s,2H),2.40(s,3H),2.20-2.10(m,2H),1.97-1.74(m,2H),1.57-1.30(m,2H),0.97-0.85(m,6H).

[0257] LC-MS (m / z): 572.2 [M+H] +

[0258] Example 11: Preparation of compound 55:

[0259]

[0260] Step 1: A solution of 13a (3.00 g, 23.781 mmol, 1 eq, purchased from Yaoshi Technology Co., Ltd.) was added to THF (30 mL). Zinc iodide (227 mg, 713.421 μmol, 0.03 eq) was added at 20 °C, followed by TMSCN (4.72 g, 47.561 mmol, 2.00 eq). After the addition was complete, the temperature was raised to 50 °C and the reaction was carried out for 5-6 hours until complete. The reaction solution was slowly poured into 300 mL of saturated NaHCO3 aqueous solution, and the pH was adjusted to 7-8. Then, MTBE (50 mL * 3) was used to combine the organic phases. The solution was washed once with 50 mL of saturated NaCl aqueous solution, dried over 10 g of anhydrous magnesium sulfate, filtered, and concentrated at 35 °C to obtain the crude product. A pale yellow liquid product 13b, 2.80 g, yield 76%, was obtained.

[0261] Step 2: Add 13b (2.80 g, 18.279 mmol, 1.00 eq) to MeOH (30 mL), and add SOCl2 (6.52 g, 54.838 mmol, 3.00 eq) dropwise at approximately 0 °C. After the addition is complete, reflux at 60-70 °C for 12 h until the reaction is complete. Dry the solvent by rotary evaporation, add 50 mL of EA and 100 mL of saturated sodium bicarbonate aqueous solution, adjust the pH to approximately 8, stir for 10 min, separate the phases, and extract the aqueous phase once more with 50 mL of EA. Combine the organic phases, dry by rotary evaporation, and perform column chromatography with an n-Heptane:EA ratio of 6:1 to 4:1. This yields 1.10 g of a brown oily substance, 13c, in 32% yield.

[0262] 1 H NMR (400MHz, CDCl3) δ4.3(s,1H),3.8(s,3H),3.3(s,3H),2.7(s,1H),1.8(s,6H)

[0263] Step 3: Add 13c (0.25g, 1.343mmol, 1.00eq) to H2O (5mL) and THF (5mL), and add LiOH at 20℃. . H₂O (84 mg, 2.014 mmol, 1.50 eq) was added and stirred for 4 h until the reactants were completely reacted. The THF was evaporated to dryness, and then 2 mL of 1 N HCl aqueous solution was added to adjust the pH to approximately 3. The solution was then evaporated to dryness again. A yellow oily product was obtained, yielding 201 mg (13 days), with a yield of 86%.

[0264] Step 4: Ecinotecan mesylate (224 mg, 422.396 μmol, 1 eq, purchased from Anqing Runke Biomedical Technology Co., Ltd.) and 13d (0.08 g, 464.635 μmol, 1.10 eq) were added to DMF (8 mL), followed by HATU (240 mg, 633.594 μmol, 1.50 eq) and DIEA (163 mg, 1.267 mmol, 3.00 eq). After the addition was complete, the reaction was continued at 20 °C for 2 h until complete. The reaction solution was directly sent to the preparation unit to obtain product 55, a pale yellow solid, 51 mg, with a yield of 20%.

[0265] 1 H NMR (400MHz, DMSO-d6) δ8.37-8.27(m,1H),7.79(d,1H,J=10.8Hz),7.31(s,1H),6.52(m,1H),5.5-5.6(m,2H),5.43(s,2H),5.2-5.3( m,2H),4.18-4.12(m,1H,),3.18-3.16(m,5H),2.40(s,3H),2.1-2.2(m,2H),1.8-1.9(m,2H),1.8-1.7(m,6H),0.88(t,3H,J=7.3Hz).

[0266] LC-MS (m / z): 590.2 [M+H] +

[0267] Example 12: Preparation of compound 56:

[0268]

[0269] Experimental steps:

[0270] Step 1: Add 12a (3.00 g, 25.832 mmol, 1.00 eq, purchased from Pharmaron Technology Co., Ltd.) to DCM (30 mL). Add Desmartin oxidant (14.24 g, 33.582 mmol, 1.30 eq) in portions at 10-20 °C. After the addition is complete, continue the reaction at 20 °C for 3-4 hours until the reaction is complete. Filter the reaction mixture directly, wash the filter cake with 30 mL of diethyl ether, and collect the filtrate to obtain a colorless liquid 12b, 2.95 g. Assuming a 100% yield, proceed directly to the next step.

[0271] Step 2: A solution of 12b (2.94 g, 25.833 mmol, 1 eq) was added to 30 mL of THF. Zinc iodide (247 mg, 774.992 μmol, 0.03 eq) was added at 20 °C, followed by TMSCN (5.12 g, 51.666 mmol, 2.00 eq). After the addition was complete, the temperature was raised to 50 °C and the reaction proceeded for 5-6 hours until complete. The reaction solution was slowly poured into 300 mL of saturated NaHCO3 aqueous solution, and the pH was adjusted to 7-8. The solution was then washed once with 50 mL of saturated NaCl aqueous solution using MTBE (50 mL x 3). The organic phases were combined, washed once with 50 mL of saturated NaCl aqueous solution, dried over 10 g of anhydrous magnesium sulfate, filtered, and concentrated at 35 °C to obtain the crude product. The resulting pale yellow liquid product, 12c, was 2.50 g in weight, with a yield of 68%.

[0272] Step 3: Add 12c (2.50 g, 17.713 mmol, 1 eq) to MeOH (25 mL), and add SOCl2 (6.322 g, 53.138 mmol, 3.00 eq) dropwise at approximately 0 °C. After the addition is complete, reflux at 60-70 °C for 12 h until the reaction is complete. Dry the solvent by rotary evaporation, add 50 mL of EA and 100 mL of saturated sodium bicarbonate aqueous solution, adjust the pH to approximately 8, stir for 10 min, separate the phases, and extract the aqueous phase once more with 50 mL of EA. Combine the organic phases, dry by rotary evaporation, and perform column chromatography with an n-Heptane:EA ratio of 6:1 to 4:1. This yields 0.95 g of a brown oily substance, 12d, in a 30% yield.

[0273] 1 H NMR (400MHz, CDCl3) δ4.42 (br d, 1H, J = 5.8Hz), 3.84 (s, 3H), 2.79 (br d, 1H, J = 6.8Hz), 2.07 (d, 6H, J = 2.5Hz).

[0274] Step 4: Add 12d (0.40g, 2.297mmol, 1.00eq) to H2O (5mL) and THF (5mL), and add LiOH at 20℃. . H₂O (144.563 mg, 3.445 mmol, 1.50 eq) was added, and the mixture was stirred for 4 hours until the reactants were completely reacted. THF was removed by rotary evaporation, and 2 mL of 1N HCl aqueous solution was added to adjust the pH to approximately 3. The solution was then evaporated to dryness. A yellow oily product, 12e, was obtained, with a yield of 310 mg and a yield of 84%.

[0275] Step 5: Ecinotecan mesylate (211 mg, 397.373 μmol, 1.00 eq, purchased from Anqing Runke Biomedical Technology Co., Ltd.) and 12e (0.07 g, 437.110 μmol, 1.10 eq) were added to DMF (10 mL), followed by HATU (226 mg, 596.060 μmol, 1.50 eq) and DIPEA (154 mg, 1.192 mmol, 3.00 eq). After the addition was complete, the reaction was continued at 20 °C for 2 h until complete. The reaction solution was directly sent to the preparation unit. Two peaks were obtained, representing the epimer, which were evaporated to dryness. The peaks were combined to obtain a pale yellow solid product of 56.85 mg, with a yield of 37%.

[0276] 1 H NMR(400MHz,DMSO-d6)δ8.46-8.35(m,1H),7.80-7.77(m,1H),7.32(s,1H),6.52(s,1H), 5.7-5.8(m,1H),5.5-5.6(m,1H),5.43(s,2H),5.2-5.3(m,2H),4.2-4.3(m,1H),3.16(br s,2H),2.40(s,3H),2.1-2.2(m,2H),2.0-2.0(m,2H),1.8-2.0(m,6H),0.88(t,3H,J=7.3Hz).

[0277] LC-MS (m / z): 578.2 [M+H] +

[0278] Example 13: Preparation of compound 57:

[0279]

[0280] Step 1: Dissolve 14a (1.00 g, 5.457 mmol, 1 eq, purchased from Pharmaron) in DMF (10 mL), then cool to 0°C, replace with nitrogen, and add NaH (436 mg, 10.900 mmol, 1.997 eq). Stir for 10 min at 0°C, then add 14b (1.57 g, 6.563 mmol, 1.203 eq, purchased from Pharmaron). The mixture is then naturally heated to 25°C and stirred for 16 h. After the reaction is complete, 10% citric acid is added to quench the reaction, followed by extraction with ethyl acetate. The organic phase is dried. Column chromatography is performed using PE:EA = 50:1. 14c, 1.50 g, is obtained as an oil, with a yield of 80%.

[0281] 1H NMR (400MHz, CDCl3) δ3.6-3.7(m,2H),3.29(t,2H,J=6.5Hz),2.38(s,1H),2.0-2.1(m,6H),1.48(s,9H),0.92(s,9H),0.08(s,6H).

[0282] Step 2: Dissolve 14c (170 mg, 497.717 μmol, 1 eq) in DCM (4 mL), add TFA (1 mL), and then incubate at 25 °C for 16 h. The reaction is complete as determined by TLC. The mixture is then evaporated to dryness and proceeded directly to the next step. 14d, 63 mg, was obtained, with a yield of 100%.

[0283] Step 3: Dissolve eczemab mesylate (100 mg, 188.128 μmol, 1 eq, purchased from Anqing Runke Biomedical Technology Co., Ltd.) in DMF (2 mL). Add DBU-CO2 complex (161 mg, 751.421 μmol, 3.994 eq), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholin-4-ium chloride (124 mg, 448.108 μmol, 2.382 eq), and 14d (57 mg, 448.174 μmol, 2.382 eq) to the solution and react at 25 °C for 3 h. After the reaction is complete, add 6 mL of water, and a solid precipitates. Filter and wash with 10 mL of water. The product was sent for preparation to give compound 57, a pale yellow solid, 7 mg, yield 6%.

[0284] 1 H NMR (400MHz, DMSO-d6) δ7.77(s,1H),7.31(s,1H),6.73(d,1H,J=8.3Hz),6.51(s,1H),5.43(s,2H),5.3-5.4(m,1H),5.1-5.3(m,2H),4.98(t ,1H,J=4.9Hz),3.4-3.5(m,2H),3.1-3.3(m,4H),2.4-2.4(m,4H),2.1- 2.3(m,2H),2.11(s,6H),1.87(q,2H,J=7.0,13.6Hz),0.8-0.9(m,3H).

[0285] LC-MS (m / z): 589.2 [M+H] +

[0286] Example 14: Preparation of compound 58:

[0287]

[0288] Step 1: Dissolve 15a (613 mg, 5.126 mmol, 1 eq, purchased from Yaoshi Technology Co., Ltd.) in DCM (27 mL), add p-anisaldehyde (697 mg, 5.119 mmol, 9.988 e-1 eq) and NaHCO3 (430 mg, 5.119 mmol, 9.986 e-1 eq), then incubate at 25°C for 4 hours, and add anhydrous magnesium sulfate. Filter, concentrate the reaction solution, add it to 4 mL of DCM, cool to 0°C, add m-CPBA (884 mg, 5.123 mmol, 9.994 e-1 eq) in dichloromethane, and react at 0°C for 1 hour, then at room temperature for 1 hour. Filter, wash with saturated sodium bicarbonate, wash with saturated brine, and dry the organic phase. The solution was evaporated to dryness, then dissolved in 6 mL of methanol. Hydroxylamine hydrochloride (712 mg, 10.246 mmol, 1.999 eq) was added, and the mixture was allowed to react at 25°C for three days. After the reaction was complete, the reaction solution was evaporated to dryness, and 5 mL of water and 6 mL of diethyl ether were added. The mixture was then extracted with diethyl ether (10 x 6 mL) to remove impurities. NaHCO3 solid was added until saturated, and then extracted again with diethyl ether (10 x 6 mL). The reaction proceeded directly to the next step without purification. A red solid 15b was obtained, with a yield of 22% and a volume of 113 mg.

[0289] 1 H NMR(400MHz, CDCl3)δ5.75(br s,2H),2.53(s,1H),1.87(s,6H)

[0290] Step 2: Dissolve eczemab mesylate (100 mg, 188.128 μmol, 1 eq, purchased from Anqing Runke Biomedical Technology Co., Ltd.) in DMF (2 mL). Add DBU-CO2 complex (161 mg, 751.421 μmol, 3.994 eq), 15b (46.059 mg, 464.631 μmol, 2.470 eq), and 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholin-4-iumchloride (130 mg, 469.791 μmol, 2.497 eq) to the solution. Then, react at 25°C for 3 hours. After the reaction is complete, add 6 mL of water, and a solid precipitates. Filter and wash with 10 mL of water. The product was sent for preparation to give compound 58, an off-white solid, 20 mg, yield 19%.

[0291] 1 H NMR (400MHz, DMSO-d6) δ9.50 (s, 1H), 7.74 (d, 1H, J = 10.9Hz), 7.64 (d, 1H, J = 9.2Hz), 7.31 (s, 1H), 6.53 (s, 1H), 5.3-5.5 (m, 3H), 5.37- 5.03(m,2H),3.2-3.3(m,1H),3.0-3.2(m,1H),2.5-2.5(m,1H),2.38(s,3H),2.1-2.2(m,8H),1.8-2.0(m,2H),0.89(t,3H,J=7.3Hz).

[0292] LC-MS (m / z): 561.1 [M+H] +

[0293] Example 15: Preparation of Compound 59

[0294]

[0295] Experimental steps:

[0296] Step 1: Dissolve eczemacin mesylate (100 mg, 188.128 μmol, 1 eq) in H2O (0.8 mL) and DMF (3.2 mL). Add 11a (55 mg, 224.241 μmol, 1.192 eq, purchased from Pharmaron Technology Co., Ltd.), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholin-4-ium chloride (104 mg, 375.833 μmol, 1.998 eq), and DIPEA (73 mg, 564.385 μmol, 3 eq) and react at 25 °C for 3 h. The reaction is complete according to TLC. Add 6 mL of water; a solid precipitates. Filter and wash with 10 mL of water. A pale yellow solid 11b, 116 mg, was obtained, with a yield of 93%.

[0297] LC-MS (m / z): 663.6 [M+H] +

[0298] Step 2: Dissolve 11b (116 mg, 175.041 μmol, 1 eq) in TFA (0.4 mL), add DCM (2 mL), and react at 20 °C for 16 h. TLC analysis showed the reaction was complete. Directly evaporate to dryness to obtain a yellow solid 11c, 98 mg, yielded 100%.

[0299] LC-MS (m / z): 563.6 [M+H] +

[0300] Step 3: Dissolve 11c (99 mg, 175.973 μmol, 1 eq) in MeOH (2 mL), add sodium cyanoborohydride (55 mg, 875.211 μmol, 4.974 eq) and 1,4-Dioxane-2,5-diol (105 mg, 874.242 μmol, 4.968 eq), displace nitrogen gas, and react at 25 °C for 16 h. TLC detection showed the reaction was complete. The reaction solution was then directly reversed to prepare a yellow solid (59 mg, 30 mg), yield 28%.

[0301] 1H NMR (400MHz, DMSO-d6) δ9.56(s,1H),9.10(d,J=8.6Hz,1H),7.83(d,J=11.0Hz,1H),7.33(s,1 H),6.55(s,1H),5.79-5.54(m,1H),5.43(s,2H),5.35-5.06(m,2H),4.39(s,1H),4.00(s,1H), 3.83-3.80(m,1H),3.75(s,2H),3.38-3.02(m,9H),2.62-2.53(m,1H),2.43(s,3H),2.32-2.2 2(m,1H),2.16-2.14(m,1H),2.01(d,J=10.1Hz,1H),1.94-1.78(m,2H),0.89(t,J=7.3Hz,3H).

[0302] LC-MS (m / z): 607.3 [M+H] +

[0303] Example 16: Preparation of compound LP-1:

[0304]

[0305] Example 17: Preparation of compound LP-2:

[0306]

[0307] Example 18: Fabrication of ADC

[0308] Take an appropriate amount of antibody into a 50 mL EP tube, slowly and evenly add 10 equivalents of TCEP solution, and place it on a shaker at 22°C (60 rpm) overnight for reaction. After 18 h, take the above solution, slowly and evenly add 10% of DMA and 12 equivalents of linker-payload (the compound LP prepared above) to the final concentration of approximately 6.0 mg / mL, and place it on a shaker at 22°C (60 rpm) for 1 h to obtain the ADC sample.

[0309] The ADC sample was concentrated in an ultrafiltration tube (Amicon) and the buffer was changed. The sample was then placed in PBS (1X), pH 7.4 buffer and centrifuged at 4200 rpm for 8 min. After three replacements, the sample was centrifuged at 4200 rpm for 8 min in a buffer of 20 mM Histidine, 8% Sucrose, 0.02% Tween 80, pH 5.5, and the buffer was changed three times. The sample was filtered through a 0.2 μm filter membrane, and the concentration was determined using Nanodrop. The concentration was detected using Nanodrop and a fluorescence detector (FLD-HPLC). The purity of the ADC sample was determined using size exclusion chromatography (SEC-HPLC). The purity and hydrophilicity / hydrophobicity of the ADC sample were determined using hydrophobic chromatography (HIC-HPLC).

[0310] Test Example 1: Compound Cytotoxicity Experiment

[0311] To investigate the cytotoxic effects of different compounds, we revived a total of eight tumor cell lines from different cancer types, including lung cancer cells (HCC827), breast cancer cells (MCF-7, T47D, MDA-MB-231, SKBR3), colorectal cancer cells (HT29, DLD-1), and gastric cancer cells (KATOⅢ), and tested the cytotoxicity of different compounds. The control compounds DXD (Cat#HY-13631D), MMAE (Cat#HY-15162), eczemab (Cat#HY-13631), SN38 (Cat#HY-13704), and Belotecan hydrochloride (Cat#HY-13566A) were all purchased from MCE.

[0312] Tumor cells were collected by digestion and resuspended in 1640 + 10% FBS (Gibco, Cat#10099-141) or DMEM + 10% FBS complete medium. Cells were counted, and the density was adjusted to 2 × 10⁻⁶ cells / mL. 3 40 μL of cells were seeded into each well of a 384-well plate. The test compound was prepared at a final detection concentration of 3-fold (maximum detection concentration of 10 μM). A 10-fold serial dilution was performed to obtain 6 or 8 concentration points, with 20 μL added to each well. After co-incubation for 4 days, 20 μL of CellCounting-Lite 2.0 (Vazyme, Cat#DD1101-02) was added to each well. After reacting at room temperature for 5 min, the luminescence intensity was measured using a multi-mode microplate reader. The percentage of target cell killing was calculated using the following formula: %Target Cell Kill = (Untreated Control Group – Test Sample Group) / Untreated Control Group.

[0313] The results are shown in the table below. Tables 1 and 2 show the kill IC. 50The values ​​are shown in Tables 3 and 4, which represent the maximum killing values. The experimental results show that the compounds of this invention have good inhibitory activity against various tumor cells. Among them, compounds 49-2, 50, 52-1, 52-2, and 53 exhibit better inhibitory activity (approximately 3-5 times) compared to DXD, and their inhibitory activity is lower than that of eczema, making them very promising toxins for ADCs with a large therapeutic window.

[0314] Table 1

[0315]

[0316] " / " indicates that it has not been tested.

[0317] Table 2

[0318]

[0319] Table 3

[0320]

[0321]

[0322] " / " indicates that it has not been tested.

[0323] Table 4

[0324]

[0325] Test Example 2: ADC Cytotoxicity Assay

[0326] To detect the cytotoxic effects of ADCs, we revived tumor cell lines with different expression levels, adjusted the cell state, and collected cells during the logarithmic growth phase for plating and detection.

[0327] Tumor cells were digested and collected, resuspended in complete tumor cell culture medium, counted, and the cell density was adjusted to 2 × 10⁶ cells / year. 3 40 μL of cells were seeded into each well of a 384-well plate. A 3-fold serial dilution of the ADC to the final detection concentration was prepared and added to each well (20 μL). After incubation at 37°C and 5% CO2 for 5 days, the cell culture plates were removed and brought to room temperature. Then, 30 μL of CellCounting-Lite 2.0 (Vazyme, Cat#DD1101-02) was added to each well, mixed well, and incubated in the dark for 5 min at room temperature. The luminescence intensity was detected using a multi-functional microplate reader (Tecan, F200 Pro). The percentage of target cell killing was calculated using the following formula: %Target Cell Kill = (Untreated Control Group – Test Sample Group) / Untreated Control Group. The experimental results show that the ADC compound of this invention has good inhibitory activity against a variety of tumor cells.

Claims

1. The compound represented by formula (I), its pharmaceutically acceptable salt, stereoisomer, solvate, or prodrug, In the formula, R1 and R2 are each independently selected from hydrogen, fluorine, chlorine, amino, nitro, hydroxyl, and methyl. R3 is selected from -CR4R5-X1R6R7, -X2R8R9, or R 10 ; X1 is selected from N or CR 11 ; X2 is selected from N or CR 12 ; R4 and R5 are each independently selected from hydrogen, halogen, hydroxyl, substituted or unsubstituted C. 1-6 Alkyl and substituted or unsubstituted C 1-6 Alkoxy; Alternatively, R4, R5, and the carbon atoms they are attached to can form substituted or unsubstituted C atoms. 3-10 Cycloalkyl and substituted or unsubstituted 3- to 10-membered heterocyclic alkyl groups; R6 and R7 are each independently selected from hydrogen, halogen, oxo group, hydroxyl group, substituted or unsubstituted C. 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 3-10 Cycloalkyl groups and substituted or unsubstituted 3- to 10-membered nitrogen-containing heterocyclic alkyl groups; Alternatively, R6, R7, and the carbon or nitrogen atom they are attached to form substituted or unsubstituted C atoms. 3-10 Cycloalkyl or substituted or unsubstituted 3- to 10-membered heterocyclic alkyl groups; R8 is selected from substituted or unsubstituted C. 3-10 Cycloalkyl and substituted or unsubstituted 3- to 10-membered heterocyclic alkyl groups; R9 is selected from hydrogen, halogen, hydroxyl, substituted or unsubstituted C. 1-6 Alkyl and substituted or unsubstituted C 1-6 Alkoxy; R 10 Selected from substituted or unsubstituted C 3-10 Cycloalkyl and substituted or unsubstituted 3- to 10-membered heterocyclic alkyl groups; R 11 R 12 Selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl and halogen-substituted C 1-6 alkyl; The term "substitution" as used herein refers to each of the aforementioned groups being independently replaced by one or more groups selected from R. A The substituents are replaced by the R; A Selected from hydrogen, deuterium, halogen, hydroxyl, oxo, cyano, nitro, C 1-6 Alkyl, C 1-6 Alkoxy group, -NHC(O)-C 1-6 Alkyl group, -NHS(O)2-C 1-6 Alkyl, C 3-10 Cycloalkyl and 3 to 10-membered heterocycloalkyl, wherein C 1-6 Alkyl, C 1-6 Alkoxy group, -NHC(O)-C 1-6 Alkyl group, -NHS(O)2-C 1-6 Alkyl, C 3-10 The cycloalkyl or 3- to 10-membered heterocycloalkyl group is optionally substituted by one or more substituents selected from halogens and hydroxyl groups; In each of the above groups, the 3 to 10-membered heterocyclic alkyl group may optionally contain 1, 2, 3 or 4 heteroatoms selected from nitrogen, oxygen or sulfur as ring atoms, and the ring atoms of the 3 to 10-membered nitrogen-containing heterocyclic alkyl group may contain at least 1 nitrogen atom and optionally also contain 1, 2 or 3 heteroatoms selected from nitrogen, oxygen or sulfur as ring atoms.

2. The compound according to claim 1, its pharmaceutically acceptable salt, stereoisomer, solvate, or prodrug, wherein, R3 contains at least one hydroxyl substituent; Preferably, R1 is methyl and R2 is fluorine; or R1 is amino and R2 is hydrogen. Preferably, R1 is methyl and R2 is fluorine.

3. The compound according to claim 1 or 2, its pharmaceutically acceptable salt, stereoisomer, solvate, or prodrug, wherein, R4 and R5 are each independently selected from hydrogen, halogen, hydroxyl, and C. 1-6 Alkyl and C 1-6 Alkoxy, the C 1-6 Alkyl and C 1-6 The alkoxy group is optionally surrounded by 1, 2, 3 or 4 groups selected from R A Substituents of the substituents; Preferably, R4 and R5 are each independently selected from hydrogen, fluorine, and chlorine; Preferably, R4 and R5 are each independently selected from hydrogen and fluorine; Preferably, R4 and R5 are both hydrogen or both R4 and R5 are fluorine; Alternatively, R4, R5, and the carbon atoms they are bonded to can form C. 3-10 cycloalkyl, the C 3-10 The cycloalkyl group is optionally surrounded by 1, 2, 3 or 4 groups selected from R A Substituents of the substituents; Preferably, R4, R5 and the carbon atoms they are attached to together form cyclopropyl, cyclobutyl, cycloalkyl or cyclohexyl; Preferably, R4, R5, and the carbon atoms they are attached to together form a cyclopropyl or cyclobutyl group; Preferably, R6 and R7 are each independently selected from hydrogen, halogen, hydroxyl, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 3-10 Cycloalkyl and 3 to 10-membered heterocycloalkyl, wherein C 1-6 Alkyl, C 1-6 Alkoxy, C 3-10 Cycloalkyl or 3 to 10-membered heterocycloalkyl groups optionally composed of 1, 2, 3 or 4 selected from R A Substituents of the substituents; Preferably, R6 and R7 are each independently selected from hydrogen, halogen, hydroxyl, and C. 1-6 Alkyl and halogen-substituted C 1-6 alkyl; Preferably, R6 and R7 are each independently selected from hydrogen, halogen, hydroxyl, and C. 1-3 Alkyl and halogen-substituted C 1-3 alkyl; Preferably, R6 and R7 are each independently selected from hydrogen, fluorine, chlorine, hydroxyl, methyl, ethyl, propyl, fluoromethyl, difluoromethyl, trifluoromethyl, fluoroethyl, difluoroethyl and trifluoroethyl; Preferably, R6 and R7 are each independently selected from hydroxyl, methyl, ethyl and trifluoromethyl; Preferably, X1 is CR 11 ;R 11 Selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl and halogen-substituted C 1-6 alkyl; Preferably, R 11 Selected from hydrogen and C 1-6 alkyl; Preferably, R 11 Selected from hydrogen and C 1-3 alkyl; Preferably, R 11 Selected from hydrogen and methyl.

4. The compound according to any one of claims 1-3, its pharmaceutically acceptable salt, stereoisomer, solvate, or prodrug, wherein, X1 is CH or N; R6, R7, and the carbon or nitrogen atoms they are connected to together form C. 3-10 Cycloalkyl or 3 to 10-membered heterocycloalkyl, wherein the C 3-10 Cycloalkyl or 3 to 10-membered heterocycloalkyl groups optionally composed of 1, 2, 3 or 4 selected from R A Substituents of the substituents; Preferably, R6, R7 and the carbon or nitrogen atom connected to them together form a 3 to 10-membered nitrogen-containing heterocyclic alkyl group, wherein the 3 to 10-membered nitrogen-containing heterocyclic alkyl group is optionally substituted by 1, 2, 3 or 4 substituents selected from hydroxyl or oxo groups; Preferably, X2 is CH or N, and R8 is selected from C. 3-10 Cycloalkyl and substituted 3- to 10-membered heterocycloalkyl, wherein C 3-10 Cycloalkyl or 3 to 10-membered heterocycloalkyl groups optionally composed of 1, 2, 3 or 4 selected from R A Substituents of the substituents; Preferably, R8 is The Optionally selected from 1, 2, 3 or 4 of R A Substituents of the substituents; Preferably, R8 is The Optionally, it is selected from 1, 2, 3 or 4 halogens or C 1-6 Substitution of alkoxy groups; Preferably, R8 is The Optionally, it is selected from 1, 2, 3 or 4 halogens or C 1-3 Substitution of alkoxy groups; Preferably, R8 is The Optionally substituted with 1, 2, 3 or 4 substituents selected from fluorine or methoxy groups; Preferably, R9 is selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl and C 1-6 alkyl-OH; Preferably, R9 is selected from hydroxyl and C. 1-6 alkyl-OH; Preferably, R9 is selected from hydroxyl and C. 1-3 alkyl-OH; Preferably, R9 is selected from hydroxyl and -(CH2)2-OH.

5. The compound according to any one of claims 1-4, its pharmaceutically acceptable salt, stereoisomer, solvate, or prodrug, wherein, R 10 Selected from C 3-10 Cycloalkyl and 3 to 10-membered heterocycloalkyl, wherein C 3-10 Cycloalkyl or 3 to 10-membered heterocycloalkyl groups optionally composed of 1, 2, 3 or 4 selected from R A Substituents of the substituents; Preferably, R 10 Selected from C 3-10 cycloalkyl, the C 3-10 The cycloalkyl group is optionally surrounded by one or two hydroxyl groups and -NHC(O)-C. 1-6 Substitution of alkyl-OH groups; Preferably, R 10 Selected from cyclopropyl, cyclobutyl, cyclopentyl, and adamantyl, wherein the cyclopropyl, cyclobutyl, cyclopentyl, or adamantyl group is optionally surrounded by one or two groups selected from hydroxyl and -NHC(O)-C. 1-6 Substitution of alkyl-OH groups; Preferably, R 10 Selected from cyclopropyl, cyclobutyl, cyclopentyl, and adamantyl, wherein the cyclopropyl, cyclobutyl, cyclopentyl, or adamantyl group is optionally surrounded by one or two groups selected from hydroxyl and -NHC(O)-C. 1-3 Substitution of alkyl-OH groups; Preferably, R 10 Selected from cyclopropyl, cyclobutyl, cyclopentyl, and adamantyl, wherein the cyclopropyl, cyclobutyl, cyclopentyl, or adamantyl group is optionally surrounded by one or two groups selected from hydroxyl and -NHC(O)-C. 1-3 Substitution of alkyl-OH groups; Preferably, R 10 Selected from cyclobutyl and adamantyl, wherein the cyclobutyl or adamantyl group is optionally substituted with a substituent selected from hydroxyl or -NHC(O)-(CH2)2-OH; Or R 10 The derivatives are selected from aziridine, pyrrolidinyl, piperidinyl, and piperazine, wherein the aziridine, pyrrolidinyl, piperidinyl, or piperazine is optionally composed of 1, 2, 3, or 4 derivatives selected from R. A Substituents of the substituents; Preferably, R 10 The group is selected from aziridine, pyrrolidinyl, piperidinyl, and piperazine, wherein the aziridine, pyrrolidinyl, piperidinyl, or piperazine is optionally surrounded by 1, 2, 3, or 4 groups selected from -C. 1-6 Alkoxy and -C 1-6 Substitution of alkyl-OH groups; Preferably, R 10 The group is selected from aziridine, pyrrolidinyl, piperidinyl, and piperazine, wherein the aziridine, pyrrolidinyl, piperidinyl, or piperazine is optionally surrounded by 1, 2, 3, or 4 groups selected from -C. 1-3 Alkoxy and -C 1-3 Substitution of alkyl-OH groups; Preferably, R 10 Selected from aziridine and pyrrolidinyl groups, wherein the aziridine or pyrrolidinyl group is optionally surrounded by 1, 2, 3 or 4 groups selected from -C 1-3 Alkoxy and -C 1-3 Substitution of alkyl-OH groups; Preferably, R 10 Selected from The Optionally substituted with one or two substituents selected from methoxy and -(CH2)2-OH; preferably, the C 3-10 Each time a cycloalkyl group appears, it is independently selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, bicyclo[1.1.0]butyl, bicyclo[1.1.1]pentyl, bicyclo[2.2.0]hexyl, bicyclo[2.1.1]hexyl, bicyclo[3.1.0]hexyl, bicyclo[2.2.1]heptyl, bicyclo[3.2.0]heptyl, bicyclo[3.2.1]octyl, bicyclo[2.2.2]octyl, bicyclo[4.2.0]octyl, bicyclo[4.3.0]nonyl, bicyclo[3.3.1]nonyl, and adamantyl; Each of the 3 to 10 membered heterocyclic alkyl groups is independently selected from tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, oxazolyl, thiazolyl, imidazolyl, piperidinyl, piperazinel, and morpholinyl when it appears. Preferably, R3 is selected from Among them, the thick solid line This indicates that the attached carbon atom exhibits chiral isomerism; Preferably, R3 is selected from 6. The compound according to claim 1, its pharmaceutically acceptable salt, stereoisomer, solvate, or prodrug, wherein, The compound of formula (I) is the same as the compound of formula (II): In the formula, R4, R5, R6, R7, and R9 are each defined as in claim 1; Preferably, R4 and R5 are hydrogen or fluorine; or R4 and R5 together with the carbon atoms they are attached to form a cyclopropyl or cyclobutyl group. Preferably, R6, R7, and R9 are each independently selected from hydroxyl, methyl, ethyl, and trifluoromethyl. Preferably, Selected from Alternatively, the compound of formula (I) may be a compound of formula (III): In the formula, R4, R5, and X1 are each defined as in claim 1; R A1 Selected from halogen, hydroxyl, oxo, cyano, nitro, C 1-6 Alkyl, C 1-6 Alkoxy group, -NHC(O)-C 1-6 Alkyl group, -NHS(O)2-C 1-6 Alkyl, C 3-10 Cycloalkyl and 3 to 10-membered heterocycloalkyl, wherein C 1-6 Alkyl, C 1-6 Alkoxy group, -NHC(O)-C 1-6 Alkyl group, -NHS(O)2-C 1-6 Alkyl, C 3-10 The cycloalkyl or 3- to 10-membered heterocycloalkyl group is optionally substituted by one or more substituents selected from halogens and hydroxyl groups; m1 and n1 are each independently selected from 0, 1, 2 and 3; Preferably, R4 and R5 are hydrogen; Preferably, X1 is N; Preferably, R A1 It is an oxo group or a hydroxyl group, n1 is 0, 1 or 2, and n1 has 1 R. A1 Same or different; Preferably, m1 is 1; Preferably, for Alternatively, the compound of formula (I) is a compound of formula (IV): In the formula, Y is NH or CH2; R A2 Selected from halogen, hydroxyl, oxo, cyano, nitro, C 1-6 Alkyl, C 1-6 Alkoxy group, -NHC(O)-C 1-6 Alkyl group, -NHS(O)2-C 1-6 Alkyl, C 3-10 Cycloalkyl and 3 to 10-membered heterocycloalkyl, wherein C 1-6 Alkyl, C 1-6 Alkoxy group, -NHC(O)-C 1-6 Alkyl group, -NHS(O)2-C 1-6 Alkyl, C 3-10 The cycloalkyl or 3- to 10-membered heterocycloalkyl group is optionally substituted by one or more substituents selected from halogens and hydroxyl groups; m2 and n2 are each independently selected from 0, 1, 2 and 3; Preferably, R A2 Selected from methoxy, -(CH2)2-OH and -NH-C(O)-CH2-OH, n2 is 0, 1 or 2, n2 R A2 Same or different; Preferably, m2 is 0 or 1; Preferably, Selected from Alternatively, the compound of formula (I) can be a compound of formula (V): In the formula, X2 is the same as in claim 1; R A3 R A4 Each is independently selected from hydrogen, halogen, hydroxyl, oxo, cyano, nitro, C 1-6 Alkyl, C 1-6 Alkoxy group, -NHC(O)-C 1-6 Alkyl group, -NHS(O)2-C 1-6 Alkyl, C 3-10 Cycloalkyl and 3 to 10-membered heterocycloalkyl, wherein C 1-6 Alkyl, C 1-6 Alkoxy group, -NHC(O)-C 1-6 Alkyl group, -NHS(O)2-C 1-6 Alkyl, C 3-10 The cycloalkyl or 3- to 10-membered heterocycloalkyl group is optionally substituted by one or more substituents selected from halogens and hydroxyl groups; Preferably, X2 is CH or N; Preferably, R A3 Selected from hydrogen, fluorine, and methoxy groups; Preferably, R A4 Selected from hydroxyl and -(CH2)2-OH; Preferably, Selected from 7. The compound according to claim 1, its pharmaceutically acceptable salt, stereoisomer, solvate, or prodrug, wherein, The compound is selected from:

8. Antibody-drug conjugates, comprising a small molecule drug, a linker, and an antibody or its antigen-binding fragment, wherein, The effective loading of the small molecule drug is the compound of any one of claims 1-7, its pharmaceutically acceptable salt, stereoisomer, solvate, or prodrug.

9. A pharmaceutical composition comprising a compound of any one of claims 1-7, a pharmaceutically acceptable salt, stereoisomer, solvate or prodrug thereof, or an antibody-drug conjugate of claim 8, and optionally a pharmaceutically acceptable carrier.

10. Use of the compound of any one of claims 1-7, its pharmaceutically acceptable salt, stereoisomer, solvate or prodrug, or the antibody-drug conjugate of claim 8 or the pharmaceutical composition of claim 9 in the preparation of an antitumor drug; Preferably, the tumor is selected from one or more of the following: liver cancer, lung cancer, colorectal cancer, pancreatic cancer, bone cancer, breast cancer, kidney cancer, esophageal cancer, ovarian cancer, oral cancer, laryngeal cancer, bile duct cancer, uterine cancer, nasal cancer, testicular cancer, meningioma, skin cancer, cervical cancer, glioma, lymphoma, leukemia, or sarcoma. Preferably, the tumor is selected from one or more of lung cancer, breast cancer, colorectal cancer, and gastric cancer; Preferably, the tumor is selected from lung cancer or breast cancer.

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