Toxin molecules suitable for antibody-drug conjugates

Abstract

The invention provides a toxin molecule suitable for an antibody-drug conjugate, and particularly provides a compound shown in the following formula (I) or pharmaceutically acceptable salt or hydrate thereof, and the compound can be used for preparing a pharmaceutical composition for treating tumor cell proliferation related diseases. # imgabs0 #

Description

Toxin molecules suitable for antibody-drug conjugates Technical Field

[0001] The present invention relates to the field of medicinal chemistry. Specifically, the present invention provides a toxin molecule having tumor cell proliferation inhibition activity. Background Art

[0002] Antibody-drug conjugates (ADCs) link monoclonal antibodies or antibody fragments to biologically active cytotoxins via stable chemical linker compounds. These ADCs leverage the specificity of antibodies for binding to surface antigens on both normal and tumor cells and the high efficacy of cytotoxic substances, while avoiding the drawbacks of the former, such as low efficacy, and the latter's excessive toxic side effects. This means that compared to traditional chemotherapy drugs, ADCs can more precisely bind to tumor cells while minimizing their effects on normal cells.

[0003] Currently, a variety of ADCs are being used in clinical trials or clinical research, such as Kadcyla, an ADC formed by combining trastuzumab and DM1 targeting Her2. There are also patents for antibodies and ADCs targeting B7H3.

[0004] There are several classes of cytotoxic small molecules used in antibody-drug conjugates (ADCs); one class is camptothecin derivatives, which exhibit anti-tumor effects by inhibiting topoisomerase I. The application of the camptothecin derivative exatecan in antibody-drug conjugates (ADCs) has been reported in the literature, but further development of more effective ADCs is still needed in this field.

[0005] DNA topoisomerases (Topo) are essential enzymes widely present in living organisms, involved in all key nuclear processes, including DNA replication, transcription, recombination, and repair. Based on the different forms of transient DNA strand breaks they induce, topoisomerases can be divided into two major categories: topoisomerase I and topoisomerase II. Topoisomerase I and II jointly catalyze the unwinding of supercoiled DNA during DNA replication, but topoisomerase II is involved in double-strand breaks, while topoisomerase I only causes single-strand breaks. Camptothecin and its analogs reversibly bind to the DNA topoisomerase I-DNA complex, forming a camptothecin-DNA topoisomerase I-DNA ternary complex, which terminates progressive unwinding and ultimately causes the replication fork to collide with the ternary complex, resulting in irreparable DNA breaks and cell death.

[0006] Summary of the Invention

[0007] The purpose of the present invention is to provide a toxin molecule suitable for antibody-drug conjugation.

[0008] The first aspect of the present invention provides a compound represented by the following formula (I), or a pharmaceutically acceptable salt or hydrate thereof:

[0009] Where n is 0 or 1;

[0010] X is selected from the following group: N or CR 0 ;

[0011] R 0 Selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, a C1-C8 alkyl group, a C1-C8 alkoxy group, OH, NH2, N3 or NO2;

[0012] R 1 Selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, a C1-C8 alkyl group, a C1-C8 alkoxy group, a C1-C8 haloalkyl group, a C1-C8 haloalkoxy group, N3, NO2, NH2, NH-OH, -NR'R", -COOR', -CONR'R", -NHR"'NR'R", wherein R', R" and R'" are each independently selected from the group consisting of hydrogen, an alkyl group, an aryl group, an arylalkyl group, an acyl group, an alkoxycarbonyl group, and an aryloxycarbonyl group;

[0013] R 2 、R 3 、R 4 、R 5 and R 6 Each is independently selected from the following groups: hydrogen atom, deuterium atom, halogen, hydroxyl, cyano, NH2, NO2, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C1-C8 alkoxy, substituted or unsubstituted C1-C8 alkylthio, substituted or unsubstituted C1-C8 deuterated alkyl, -(CH2) m tri(C1-C4alkyl)silyl, -(CH2) m (C3-C8 cycloalkyl), -(CH2) m (3-12 membered heterocyclic group), -(CH2) m N(R 7 )2、-(CH2) m S(CH2) p R 7 、-(CH2) m S(O)(CH2) p R 7 、-(CH2) m S(O)2(CH2) p R 7 、-(CH2) m NH(CH2) p R 7 、-(CH2)m NHC(O)(CH2) p R 7 、-(CH2) m OC(O)(CH2) p R 7 、-(CH2) m C(O)(CH2) p R 7 , -CH=N(OtBu); wherein m and p are each independently 0, 1, 2, 3 or 4;

[0014] Or, R 2 and R 3 Together with the carbon atom to which it is attached, it forms a substituted or unsubstituted C5-C8 carbocyclic ring or a substituted or unsubstituted 5-12 membered heterocyclic ring;

[0015] Or, R 2 and R 3 The carbon atoms connected thereto together form a structure selected from the group consisting of: unsubstituted or substituted with one or more R a substituted saturated or unsaturated 5-6 membered carbon ring, unsubstituted or replaced by one or more R a a substituted saturated or unsaturated 5-6 membered heterocycle;

[0016] Or, R 3 and R 4 , or R 4 and R 5 The carbon atoms connected thereto together form a structure selected from the group consisting of: unsubstituted or substituted with one or more R a substituted saturated or unsaturated 5-12 membered carbon ring, unsubstituted or replaced by one or more R a substituted saturated or unsaturated 5-12 membered heterocycle; said R a is a substituted or unsubstituted substituent selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, a nitrile group, a nitro group, a hydroxyl group, an amino group, a C1-C6 alkyl group-NH-, a (C1-C6 alkyl)2N-, a C1-C6 alkyl group, a C2-C6 alkenyl group, a C2-C6 alkynyl group, a C1-C6 alkoxy group, an allyl group, a benzyl group, a C6-C 12 Aryl, C1-C6 alkoxy-C1-C6 alkyl, C1-C6 alkoxy-carbonyl, phenoxycarbonyl, C2-C6 alkynyl-carbonyl, C2-C6 alkenyl-carbonyl, C3-C6 cycloalkyl-carbonyl, C1-C6 alkyl-sulfonyl, phenyl, 5-7 membered heteroaryl, C3-C8 cycloalkyl, 3-12 membered heterocyclyl, -(CH2) m N(R 7 )2、-(CH2) m S(CH2) p R 7 、-(CH2)m S(O)(CH2) p R 7 、-(CH2) m S(O)2(CH2) p R 7 、-(CH2) m NH(CH2) p R 7 、-(CH2) m NHC(O)(CH2) p R 7 、-(CH2) m OC(O)(CH2) p R 7 、-(CH2) m C(O)(CH2) p R 7 ; wherein m and p are each independently 0, 1, 2, 3 or 4, preferably 0, 1 or 2;

[0017] Each R 7 Each is independently selected from the following group: a hydrogen atom, a deuterium atom, a halogen, a substituted or unsubstituted C1-C8 alkyl group, a C1-C8 haloalkyl group, a C1-C8 deuterated alkyl group, a substituted or unsubstituted C1-C8 alkoxy group, a hydroxyl group, an amino group, a cyano group, a nitro group, a mercapto group, a substituted or unsubstituted C1-C8 alkylene group-OH, a substituted or unsubstituted C1-C8 alkylene group-NH2, SO2Me, -OC(O)(substituted or unsubstituted C1-C4 alkyl group), -C(O)(substituted or unsubstituted C1-C4 alkyl group), a substituted or unsubstituted phenyl group, a substituted or unsubstituted 5-7 membered heteroaryl group, a substituted or unsubstituted C3-C8 cycloalkyl group, a substituted or unsubstituted 3-12 membered heterocyclyl group;

[0018] Unless otherwise specified, each of the above groups may be substituted by a substituent selected from the following groups: deuterium atom, halogen, nitrile, nitro, hydroxyl, amino, C1-C6 alkyl-NH-, (C1-C6 alkyl)2N-, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C2-C6 alkenyl, halogenated C2-C6 alkynyl, halogenated C1-C6 alkoxy, allyl, benzyl, C6-C 12 Aryl, C1-C6 alkoxy-C1-C6 alkyl, C1-C6 alkoxy-carbonyl, phenoxycarbonyl, C2-C6 alkynyl-carbonyl, C2-C6 alkenyl-carbonyl, C3-C6 cycloalkyl-carbonyl, C1-C6 alkyl-sulfonyl, phenyl, 5-7 membered heteroaryl, C3-C8 cycloalkyl, 3-12 membered heterocyclyl; and the compound is not a structure selected from the following groups:

[0019] 、

[0020] In another preferred embodiment, the compound of formula I has a structure shown in formula II or formula III below:

[0021] In another preferred embodiment, the compound of formula I has a structure shown in the following formula IV or formula V:

[0022] In another preferred embodiment, the compound of formula I has a structure shown in the following formula VI or formula VII:

[0023] In another preferred embodiment, it is characterized in that the R 4 Selected from the following groups: hydrogen atom, deuterium atom, halogen, hydroxyl, cyano, NH2, NO2, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C1-C8 alkoxy, substituted or unsubstituted C1-C8 alkylthio, substituted or unsubstituted C1-C8 deuterated alkyl, -(CH2) m (C3-C8 cycloalkyl), -(CH2) m (3-12 membered heterocyclic group), -(CH2) m N(R 7 )2、-(CH2) m S(O)(CH2) p R 7 、-(CH2) m S(O)2(CH2) p R 7 、-(CH2) m NH(CH2) p R 7 ; wherein m and p are each independently 0, 1 or 2, R 7 is defined as above;

[0024] R 5 Selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, NH2, OH, a substituted or unsubstituted C1-C8 alkyl group, a substituted or unsubstituted C1-C8 alkoxy group;

[0025] Or, R 4 and R 5 The carbon atoms connected thereto together form a structure selected from the group consisting of: unsubstituted or substituted with one or more R a substituted saturated or unsaturated 5-6 membered carbon ring, unsubstituted or replaced by one or more R a Substituted saturated or unsaturated 5-6 membered heterocyclic ring; wherein R aThe definition of is as described above.

[0026] In another preferred embodiment, R 4 Selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, a hydroxyl group, an amino group, a C1-C3 alkyl group, a C1-C3 alkoxy group, a deuterated C1-C3 alkyl group, a deuterated C1-C3 alkoxy group, a halogen-substituted C1-C3 alkyl group, a halogen-substituted C1-C3 alkoxy group, a C2-C4 alkynyl group, and a C3-C6 cycloalkyl group;

[0027] In another preferred embodiment, R 5 Selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, a hydroxyl group, an amino group, a C1-C3 alkyl group, a C1-C3 alkoxy group, a deuterated C1-C3 alkyl group, a deuterated C1-C3 alkoxy group, a halogen-substituted C1-C3 alkyl group, and a halogen-substituted C1-C3 alkoxy group;

[0028] In another preferred embodiment, R 4 Selected from the group consisting of methyl, ethyl, methoxy, deuterated methoxy, monofluoromethyl, difluoromethyl, trifluoromethyl, monofluoromethoxy, difluoromethoxy, trifluoromethoxy, ethynyl, and cyclopropyl;

[0029] In another preferred embodiment, R 5 Selected from the group consisting of methyl, ethyl, methoxy, deuterated methoxy, monofluoromethyl, difluoromethyl, trifluoromethyl, monofluoromethoxy, difluoromethoxy, trifluoromethoxy;

[0030] In another preferred embodiment, R 4 and R 5 They are connected to form substituted or unsubstituted -OCH2O-, or substituted or unsubstituted -O(CH2)2O-.

[0031] In another preferred embodiment, R 4 and R 5 They are connected to form -OCH2O-, -OCF2O- or -O(CH2)2O-.

[0032] In another preferred embodiment, the compound of formula I has the structure shown in the following formula VIII:

[0033] Wherein, ring A is substituted or replaced by one or more R a substituted saturated or unsaturated 5-6 membered carbon ring, unsubstituted or replaced by one or more R a Substituted saturated or unsaturated 5-6 membered heterocyclic ring; wherein R a The definition of R is as described above. 1 、R 2 、R 3 、R 6 , X is defined as described above.

[0034] In another preferred embodiment, the compound of formula I has the structure shown in the following formula IX:

[0035] In another preferred embodiment, the compound of formula I has the structure shown in the following formula X-XV:

[0036] Wherein, Ring A may be unsubstituted or optionally substituted; wherein, the definition of the substituent is as described above.

[0037] In another preferred embodiment, R 2 and R 3 are each independently selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, NH2, a substituted or unsubstituted C1-C8 alkyl group, a substituted or unsubstituted C1-C8 deuterated alkyl group, -(CH2) m (C3-C6 cycloalkyl), -(CH2) m (3-6 membered heterocyclic group), -(CH2) m N(R 7 )2、-(CH2) m OC(O)R 7 ; wherein m is 0, 1, 2, 3 or 4, R 7 is defined as above;

[0038] Or, R 2 and R 3 The carbon atoms connected thereto together form a structure selected from the group consisting of: unsubstituted or substituted with one or more R a substituted saturated or unsaturated 5-6 membered carbon ring, unsubstituted or replaced by one or more R a Substituted saturated or unsaturated 5-6 membered heterocyclic ring; wherein R a The definition of is as described above.

[0039] In another preferred embodiment, the compound of formula I has a structure shown in the following formula XVI:

[0040] R 2 and R 3 The carbon atoms connected thereto together form a structure selected from the group consisting of: unsubstituted or substituted with one or more R a substituted saturated or unsaturated 5-6 membered carbon ring, unsubstituted or replaced by one or more R a Substituted saturated or unsaturated 5-6 membered heterocyclic ring; wherein R a The definition of is as described above.

[0041] In another preferred embodiment, the compound of formula I has a structure shown in the following formula XVII:

[0042] In another preferred embodiment, the compound of formula I has a structure shown in the following formula XVIII-X:

[0043] Ring B may be unsubstituted or optionally substituted; wherein the substituents are as defined above.

[0044] In another preferred embodiment, the ring B is substituted by a substituent as shown below: -L x -R 11

[0045] Wherein, x is 1, 2, 3, 4, 5 or 6;

[0046] Each L is independently selected from the group consisting of CH2, O, S, NH, NHC(O), C(O), C(NH), S(O), S(O)2, C6-C 10 aryl, 5-10 membered heteroaryl, 3-8 membered saturated or unsaturated carbocyclic ring, 4-10 membered saturated or unsaturated heterocyclic ring;

[0047] R 11 Selected from the group consisting of OH, SH, NH2.

[0048] In another preferred embodiment, ring B may be unsubstituted or replaced by -L x -R 11 Any replacement;

[0049] Wherein, x is any integer from 0 to 10;

[0050] Each L is independently selected from the group consisting of: CR w R x , O, S, NH, NR y ,NHC(O),C(O),C(NH),S(O),S(O)2,C 6- C 10 aryl, 5-10 membered heteroaryl, 3-8 membered saturated or unsaturated carbocyclic ring, 3-10 membered saturated or unsaturated heterocyclic ring;

[0051] R 11 Selected from the following group: hydrogen atom, OH, SH, NH2, NHR z .

[0052] The R w 、R x 、R y and R z are independently selected from the group consisting of hydrogen atoms, deuterium atoms, halogens, hydroxyl groups, NH2, substituted or unsubstituted C 1-4 Alkyl, C 1-4Halogenated alkyl, C 4- C 10 Cycloalkylalkyl, substituted or unsubstituted C 3- C6 cycloalkyl.

[0053] In another preferred embodiment, the R a Selected from the following groups: hydrogen atom, deuterium atom, halogen, nitrile group, nitro group, hydroxyl group, amino group, C1-C6 alkyl-NH-, (C1-C6 alkyl) 2N-, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C2-C6 alkenyl, halogenated C2-C6 alkynyl, halogenated C1-C6 alkoxy, allyl, benzyl, C6-C 12 Aryl, C1-C6 alkoxy-C1-C6 alkyl, C1-C6 alkoxy-carbonyl, -(CH2) m N(R 7 )2、-(CH2) m NH(CH2) p R 7 、-(CH2) m NHC(O)(CH2) p R 7 ; wherein m and p are each independently 0, 1, 2, 3 or 4, preferably 0, 1 or 2.

[0054] In another preferred embodiment, R 1 and R 6 are each independently a hydrogen atom.

[0055] In another preferred embodiment, X is independently selected from N and CH.

[0056] In another preferred embodiment, X is independently selected from CH.

[0057] In another preferred embodiment, the R 4 and R 5 Each is independently selected from the following group: a hydrogen atom, a deuterium atom, a halogen, a hydroxyl group, NH2, a substituted or unsubstituted C1-C4 alkyl group, a substituted or unsubstituted C1-C4 alkoxy group, and a cyclopropyl group;

[0058] Or, R 4 and R 5 The carbon atoms to which it is attached together form an unsubstituted or substituted carbon atom or carbon atoms. a Substituted oxa 5-6 membered heterocycle; wherein R a The definition of is as described above.

[0059] In another preferred embodiment, R 2Selected from the group consisting of: hydrogen atom, deuterium atom, halogen, NH2, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C1-C8 deuterated alkyl, -(CH2) m (C3-C6 cycloalkyl), -(CH2) m (3-6 membered heterocyclic group), -(CH2) m N(R 7 )2、-(CH2) m OC(O)R 7 ; wherein m is 0, 1, 2, 3 or 4;

[0060] R 3 Each is selected from the following groups: hydrogen atom, deuterium atom, halogen, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C1-C8 deuterated alkyl, -(CH2) m (C3-C6 cycloalkyl), -(CH2) m (3-6 membered heterocyclic group), -(CH2) m N(R 7 )2、-(CH2) m OC(O)R 7 ; wherein m is 0, 1, 2, 3 or 4;

[0061] Or, R 2 and R 3 The carbon atoms connected thereto together form a structure selected from the group consisting of: unsubstituted or substituted with one or more R a substituted saturated or unsaturated 5-6 membered carbon ring, unsubstituted or replaced by one or more R a a substituted saturated or unsaturated 5-6 membered heterocycle;

[0062] R 4 Selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, a hydroxyl group, a cyano group, NH2, NO2, a substituted or unsubstituted C1-C8 alkyl group, a substituted or unsubstituted C1-C8 alkoxy group;

[0063] R 5 Selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, and a substituted or unsubstituted C1-C8 alkyl group;

[0064] Or, R 4 and R 5 connected to form unsubstituted or one or more R a A substituted structure selected from the group consisting of: -OCH2O- or -O(CH2)2O-;

[0065] R 7 Selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, a substituted or unsubstituted C1-C8 alkyl group, a hydroxyl group, an amino group, a cyano group, a nitro group, and a mercapto group;

[0066] Among them, R a The definition of is as described above.

[0067] In another preferred embodiment, the compound has a structure shown in the following formula:

[0068] The second aspect of the present invention provides a pharmaceutical composition comprising the compound of formula I according to any one of the first aspect of the present invention, or a pharmaceutically acceptable salt or hydrate thereof, and one or more pharmaceutically acceptable excipients, diluents or carriers.

[0069] The third aspect of the present invention provides a use of the compound of formula I as described in the first aspect of the present invention for preparing a pharmaceutical composition for treating diseases associated with tumor cell proliferation.

[0070] In another preferred embodiment, the disease is selected from the following group: breast cancer, ovarian cancer, cervical cancer, lung cancer, uterine cancer, prostate cancer, kidney cancer, urethral cancer, bladder cancer, liver cancer, gastric cancer, endometrial cancer, salivary gland cancer, esophageal cancer, melanoma, glioma, neuroblastoma, sarcoma, pharyngeal cancer, colon cancer, rectal cancer, colorectal cancer, leukemia, bone cancer, skin cancer, thyroid cancer, pancreatic cancer, lymphoma.

[0071] The fourth aspect of the present invention provides a use of the compound of formula I as described in the first aspect of the present invention, characterized in that it is used as a toxin molecule to prepare a conjugated drug, and the conjugated drug includes: antibody conjugated drug, polypeptide drug conjugate, small molecule conjugated drug, polymer conjugated drug, lipid drug conjugate and protein drug conjugate.

[0072] In a fifth aspect of the present invention, there is provided an intermediate compound as shown in the following formula:

[0073] Wherein, the definitions of each group are as described above.

[0074] In another preferred embodiment, the compound has the structure described in Formula II-b or III-b:

[0075] In another preferred embodiment, the compound has the structure described in Formula IV-b or Vb:

[0076] The sixth aspect of the present invention provides a method for preparing the compound of formula I according to the first aspect of the present invention, the method comprising the steps of:

[0077] In an inert solvent, the compound of formula Ia is reacted with the compound of formula Ib to obtain the compound of formula I.

[0078] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. DETAILED DESCRIPTION

[0079] After extensive and intensive research, the present inventors unexpectedly discovered a compound represented by Formula I. This compound exhibits unexpected activity in inhibiting tumor cell proliferation and is useful for treating diseases associated with tumor cell proliferation. Based on this discovery, the present invention was completed.

[0080] definition

[0081] As used herein, the term "alkyl" includes straight or branched chain alkyl groups. For example, C1-C8 alkyl groups represent straight or branched chain alkyl groups having 1 to 8 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, and the like.

[0082] As used herein, the term "alkenyl" includes straight or branched alkenyl groups. For example, C2-C6 alkenyl refers to a straight or branched alkenyl group having 2 to 6 carbon atoms, such as vinyl, allyl, 1-propenyl, isopropenyl, 1-butenyl, 2-butenyl, or the like.

[0083] As used herein, the term "alkynyl" includes straight or branched chain alkynyl groups. For example, C2-C6 alkynyl refers to a straight or branched chain alkynyl group having 2 to 6 carbon atoms, such as ethynyl, propynyl, butynyl, or the like.

[0084] As used herein, the term "C3-C 10 "Cycloalkyl" refers to a cyclic alkyl group having 3 to 10 carbon atoms. It may be a monocyclic ring, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or the like. It may also be a bicyclic ring, such as a bridged ring or a spiro ring.

[0085] As used herein, the term "C1-C8 alkylamino" refers to an amino group substituted by a C1-C8 alkyl group, which may be monosubstituted or disubstituted; for example, methylamino, ethylamino, propylamino, isopropylamino, butylamino, isobutylamino, tert-butylamino, dimethylamino, diethylamino, dipropylamino, diisopropylamino, dibutylamino, diisobutylamino, di-tert-butylamino, etc.

[0086] As used herein, the term "C1-C8 alkoxy" refers to a straight or branched alkoxy group having 1 to 8 carbon atoms; for example, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tert-butoxy, etc.

[0087] As used herein, the term "3-10 membered heterocycloalkyl group having 1-3 heteroatoms selected from the group consisting of N, S, and O" refers to a saturated or partially saturated cyclic group having 3-10 atoms, 1-3 of which are heteroatoms selected from the group consisting of N, S, and O. It may be a monocyclic or bicyclic ring, such as a bridged ring or a spirocyclic ring. Specific examples include oxetane, azetidine, tetrahydro-2H-pyranyl, piperidinyl, tetrahydrofuranyl, morpholinyl, and pyrrolidinyl.

[0088] As used herein, the term "C6-C 10 The term "aryl" refers to an aromatic group having 6 to 10 carbon atoms, for example, phenyl or naphthyl and the like.

[0089] As used herein, the term "5-10 membered heteroaryl having 1-3 heteroatoms selected from the group consisting of N, S, and O" refers to a cyclic aromatic group having 5-10 atoms, 1-3 of which are heteroatoms selected from the group consisting of N, S, and O. It may be a monocyclic or condensed ring. Specific examples include pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, pyrrolyl, pyrazolyl, imidazolyl, (1,2,3)-triazolyl and (1,2,4)-triazolyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, and oxazolyl.

[0090] Unless otherwise specified as "substituted or unsubstituted", the groups of the present invention may be substituted by substituents selected from the group consisting of halogen, nitrile, nitro, hydroxyl, amino, C1-C6 alkyl-amino, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C2-C6 alkenyl, halogenated C2-C6 alkynyl, halogenated C1-C6 alkoxy, allyl, benzyl, C6-C 12 Aryl, C1-C6 alkoxy-C1-C6 alkyl, C1-C6 alkoxy-carbonyl, phenoxycarbonyl, C2-C6 alkynyl-carbonyl, C2-C6 alkenyl-carbonyl, C3-C6 cycloalkyl-carbonyl, C1-C6 alkyl-sulfonyl, etc.

[0091] As used herein, "halogen" or "halogen atom" refers to F, Cl, Br, and I. More preferably, the halogen or halogen atom is selected from F, Cl, and Br. "Halogenated" means substituted with an atom selected from F, Cl, Br, and I.

[0092] Unless otherwise specified, the structural formulas described herein are intended to include all isomeric forms (e.g., enantiomers, diastereomers, and geometric isomers (or conformers)): for example, R and S configurations containing asymmetric centers, (Z) and (E) isomers of double bonds, etc. Therefore, single stereochemical isomers of the compounds of the present invention or mixtures of their enantiomers, diastereomers, or geometric isomers (or conformers) are all within the scope of the present invention.

[0093] As used herein, the term "tautomer" refers to structural isomers of different energies that can interconvert across a low energy barrier. For example, proton tautomers (i.e., prototropy) include interconversion via proton migration, such as 1H-indazole and 2H-indazole. Valence tautomers include interconversion via reorganization of some of the bonding electrons.

[0094] As used herein, the term "hydrate" refers to a complex formed by coordination of a compound of the present invention with water.

[0095] The compounds of the present application can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining specific embodiments with other chemical synthesis methods, and equivalent replacement methods well known to those skilled in the art. Preferred embodiments include but are not limited to the examples of the present application.

[0096] The solvents used in this application can be obtained commercially, and the compounds can be artificially or The software named the commercially available compounds using the supplier's catalog name.

[0097] Pharmaceutical compositions and methods of administration

[0098] Since the compounds of the present invention have excellent tumor cell proliferation inhibitory activity, the compounds of the present invention and their various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used to prevent and / or treat (stabilize, alleviate or cure) diseases associated with tumor cell proliferation.

[0099] The pharmaceutical compositions of the present invention comprise a safe and effective amount of a compound of the present invention and a pharmaceutically acceptable excipient or carrier. "Safe and effective amount" means an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, more preferably 1-200 mg per dose. Preferably, "one dose" is one capsule or tablet.

[0100] "Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compound of the present invention and with each other without significantly reducing the efficacy of the compound. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as Tween ), wetting agents (such as sodium lauryl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0101] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administration methods include (but are not limited to): oral administration, parenteral administration (intravenous administration, intramuscular administration, or subcutaneous administration).

[0102] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0103] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compounds in such compositions can be delayed in a certain portion of the digestive tract. Examples of useful encapsulating components are polymeric substances and waxes. If desired, the active compound can also be microencapsulated with one or more of the above-mentioned excipients.

[0104] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, or mixtures thereof.

[0105] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0106] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0107] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0108] The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable therapeutic agents.

[0109] When administered in combination, the pharmaceutical composition may further comprise one or more (2, 3, 4, or more) other pharmaceutically acceptable therapeutic agents. One or more (2, 3, 4, or more) of the other pharmaceutically acceptable therapeutic agents may be used simultaneously, separately, or sequentially with the compound of the present invention to prevent and / or treat diseases associated with tumor cell proliferation.

[0110] When using a pharmaceutical composition, a safe and effective amount of the compound of the present invention is administered to a mammal (e.g., a human) in need of treatment, wherein the dosage is a pharmaceutically effective dosage. For a 60 kg human, the daily dosage is generally 1 to 2000 mg, preferably 1 to 500 mg. Of course, the specific dosage will also take into account factors such as the route of administration and the patient's health condition, all of which are within the skill of a skilled physician.

[0111] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0112] Example 1

[0113] first step

[0114] 1a (1.95 g, 7.40 mmol) and imidazole (2.52 g, 37.00 mmol) were placed in a three-necked flask (100 mL). After nitrogen substitution, anhydrous N,N-dimethylformamide (30 mL) was added to the flask. The mixture was cooled to 0°C, and triethylchlorosilane (4.45 g, 29.60 mmol) and 4-dimethylaminopyridine (0.90 g, 7.40 mmol) were added sequentially. After complete addition, the mixture was stirred at 0°C for 2 hours. The reaction solution was diluted with ethyl acetate (200 mL), and the organic phase was washed with saturated brine (25 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to obtain 1b-1 (0.90 g) and 1b-2 (1.50 g) in a total yield of 73%.

[0115] 1b-1:

[0116] MS-ESI calculated value [M+H] + 378, the actual measured value is 378.

[0117] 1b-2:

[0118] MS-ESI calculated value [M+H] + 492, the measured value is 378 (one molecule of TES).

[0119] Step 2

[0120] 1b-2 (1.10 g, 2.20 mmol) and Lawesson's reagent (1.80 g, 4.40 mmol) were placed in a three-necked flask (100 mL) in sequence. After nitrogen displacement, anhydrous toluene (30 mL) was added to the three-necked flask, and the mixture was heated to 90°C and stirred for 5-6 hours. The reaction solution was diluted with ethyl acetate (200 mL), and the organic phase was washed with saturated brine (25 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to obtain 1c (620 mg) in a yield of 51%.

[0121] MS-ESI calculated value [M+H] + 394, the actual measured value is 394.

[0122] Step 3

[0123] Compound 1c (540 mg, 1.37 mmol) was dissolved in tetrahydrofuran (20 mL). Triethylamine trihydrofluoride (552 mg, 3.43 mmol) was added under ice-cooling. After complete addition, the mixture was slowly returned to room temperature and stirred overnight. The reaction solution was diluted with ethyl acetate (100 mL). The organic phase was washed with saturated brine (25 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to obtain 1 (210 mg) in a yield of 55%.

[0124] MS-ESI calculated value [M+H] + 280, the actual measured value is 280.

[0125] 1 H NMR (500MHz, CDCl3) δ7.56 (s, 1H), 6.12 (d, J = 17.5Hz, 1H), 5.25 (d, J = 17.5Hz, 1H), 4.71–4.52(m,2H),3.09–3.03(m,2H),1.80(q,J=7.0Hz,2H),0.96(t,J=7.0Hz,3H).

[0126] Example 2

[0127] first step

[0128] Under nitrogen atmosphere, 1.0M boron trichloride n-heptane solution (94mL, 94.00mmol) was added to a dried three-necked flask (500mL), and anhydrous 1,2-dichloroethane (150mL) was added. The mixed solution was cooled to 0℃, and 2a (9.00g, 72.00mmol) was added in batches. After stirring at 0℃ for ten minutes, chloroacetonitrile (7.05g, 94.00mmol) and aluminum trichloride (13.43g, 101.00mmol) were added. After the addition was complete, the mixture was slowly heated to 80℃ and refluxed for 40 hours. The reaction system was cooled to 0℃, and ice water (50mL) and hydrochloric acid aqueous solution (1N, 100mL) were slowly added in sequence. After stirring at 0℃ for half an hour, the liquid was separated and the aqueous phase was washed with dichloromethane / methanol mixed solvent (V 二氯 甲烷 :V 甲醇 =4:1, 200 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to give a crude product 2b (5.5 g), which was used directly in the next step.

[0129] MS-ESI calculated value [M+H] + 202, the actual measured value is 202.

[0130] Step 2

[0131] Under a nitrogen atmosphere, the crude product 2b (700 mg) from the previous step was dissolved in anhydrous dichloromethane (5 mL), and benzylamine (3 mL) was added. The mixture was stirred at room temperature for 5 hours. The reaction solution was directly concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to obtain compound 2c (390 mg) in a yield of 41%.

[0132] MS-ESI calculated value [M+H] + 273, the measured value is 273.

[0133] Step 3

[0134] Compound 2c (390 mg, 1.43 mmol) and 10% wet palladium on carbon (90 mg) were added sequentially to a single-necked flask (100 mL). Anhydrous tetrahydrofuran (10 mL) was added, and the hydrogen atmosphere was replaced with a hydrogen balloon. The mixture was stirred at room temperature under a hydrogen atmosphere for 20 hours. The reaction system was filtered to remove insoluble matter, and the filtrate was concentrated under reduced pressure to obtain the crude product 2d (290 mg).

[0135] MS-ESI calculated value [M+H] + 183, the actual measured value is 183.

[0136] Step 4

[0137] The crude product 2d (290 mg) from the previous step was dissolved in a tetrahydrofuran / water mixed solvent (V 四氢呋喃 :V 水 =1:1, 10 mL), 9-fluorenylmethyl chloroformate (410 mg, 1.59 mmol) and potassium carbonate (439 mg, 3.18 mmol) were added, and the mixture was stirred at 0°C for half an hour. The reaction system was diluted with ethyl acetate (100 mL), and the organic phase was washed with saturated brine (25 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to obtain compound 2e (390 mg), with a two-step yield of 67%.

[0138] MS-ESI calculated value [M+H] + 405, the actual measured value is 405.

[0139] Step 5

[0140] To a 100 mL Schlenk reaction tube were added 2e (240 mg, 0.59 mmol), 1 (165 mg, 0.59 mmol), and p-toluenesulfonic acid (60 mg, 0.35 mmol) in sequence. Anhydrous toluene (6 mL) was added, and the mixture was heated to 110°C with stirring for 2 hours. The reaction solution was concentrated under reduced pressure to obtain the crude product, which was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to afford compound 2f (120 mg) in a 31% yield.

[0141] MS-ESI calculated value [M+H] + 648, the actual measured value is 648.

[0142] Step 6

[0143] 2f (120 mg, 0.19 mmol) was dissolved in anhydrous tetrahydrofuran (2.5 mL), and diethylamine (135 mg, 1.85 mmol) was added. The mixture was stirred at room temperature for 5 hours. The reaction system was concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 2 (30 mg) in a yield of 38%.

[0144] MS-ESI calculated value [M+H] + 426, the actual measured value is 426.

[0145] 1 H NMR (400MHz, DMSO-d6) δ8.30(d,J=8.4Hz,1H),7.91(d,J=10.8Hz,1H),7.80(s,1H),6.69(s,1H),5.93(d,J=16.4 Hz,1H),5.71(s,2H),5.51(d,J=16.8Hz,1H),4.41(s,2H),2.51(s,3H),1.96–1.85(m,2H),0.86(t,J=7.2Hz,3H).

[0146] Example 3

[0147] first step

[0148] To a single-necked flask (25 mL) were added 2 (18 mg, 0.04 mmol) and glycolic acid (6 mg, 0.08 mmol) in sequence, followed by N,N-dimethylformamide (1.5 mL). The reaction system was cooled to 0°C, and N,N-diisopropylethylamine (16 mg, 0.12 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (15 mg, 0.04 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was diluted with ethyl acetate (100 mL), and the organic phase was washed with saturated brine (25 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by preparative thin-layer chromatography (methanol:dichloromethane) to obtain compound 3 (4.4 mg) in a yield of 21%.

[0149] MS-ESI calculated value [M+H] + 484, the actual measured value is 484.

[0150] 1 H NMR (400MHz, DMSO-d6) δ8.80(t,J=6.0Hz,1H),8.50(d,J=8.0Hz,1H),7.94(d,J=10.8Hz,1H),7.80(s,1H),6.68(s,1H),5.93(d,J=16.8 Hz,1H),5.78(s,2H),5.52(d,J=16.4Hz,1H),4.89(d,J=6.0Hz,2H),3.86(s,2H),2.51(s,3H),1.95–1.83(m,2H),0.86(t,J=7.2Hz,3H).

[0151] Example 4

[0152] first step

[0153] Under nitrogen atmosphere, 1.0M boron trichloride n-heptane solution (12.8mL, 12.80mmol) was added to a dried three-necked flask (100mL), anhydrous 1,2-dichloroethane (50mL) was added, the mixed solution was cooled to 0℃, 2a (2.00g, 16.00mmol) was added in batches, and 5-bromovaleronitrile (3.63g, 22.40mmol) and aluminum trichloride (3.38g, 22.40mmol) were added after stirring at 0℃ for ten minutes. After the addition was complete, the mixed system was slowly heated to 80℃ and refluxed for 30 hours. The reaction system was cooled to 0℃, ice water (50mL) and hydrochloric acid aqueous solution (1N, 60mL) were slowly added in sequence, and stirring was continued at 0℃ for half an hour before separation. The aqueous phase was washed with dichloromethane / methanol mixed solvent (V 二氯甲烷 :V 甲醇 =4:1, 200 mL) and the organic phases were combined and dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (ethyl acetate:petroleum ether=0-100%) to give crude product 4a (3.1 g), which was used directly in the next step.

[0154] MS-ESI calculated value [M+H] + 288, 290, the actual measured values ​​are 288, 290.

[0155] Step 2

[0156] To a 100 mL Schlenk reaction tube, 4a (495 mg), 1 (320 mg, 1.15 mmol), and p-toluenesulfonic acid (198 mg, 1.15 mmol) were added sequentially. Anhydrous toluene (6 mL) was added, and the mixture was heated to 110°C with stirring for 2 hours. The reaction solution was concentrated under reduced pressure to obtain the crude product, which was then purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to afford crude product 4b (200 mg).

[0157] MS-ESI calculated value [M+H] + 531, 533, the measured values ​​are 531, 533.

[0158] Step 3

[0159] 4b (90 mg) was dissolved in hexamethylphosphoric triamide (0.9 mL) and water (0.1 mL). The mixture was heated to 101°C and stirred for 4 hours. The reaction system was diluted with ethyl acetate (100 mL). The organic phase was washed with saturated brine (25 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was separated and purified by preparative high-performance liquid chromatography to obtain compound 4 (12 mg) in a yield of 15%.

[0160] MS-ESI calculated value [M+H] + 469, the actual measured value is 469.

[0161] 1H NMR (400MHz, DMSO-d6) δ8.26(d,J=8.0Hz,1H),7.90(d,J=10.8Hz,1H),7.79(s,1H),5.92(d,J=16.4Hz,1H),5.78(s,2H),5.50(s,2H),5 .49(d,J=16.4Hz,1H),3.29–3.20(m,2H),2.51(s,3H),1.94–1.82(m,2H),1.82–1.70(m,2H),1.67–1.56(m,2H),0.86(t,J=7.2Hz,3H).

[0162] Example 5

[0163] first step

[0164] 5a (2.50 g, 17.00 mmol) was dissolved in anhydrous dichloromethane (25 mL), cooled to 0°C, and triethylamine (2.60 g, 20.00 mmol) and acetyl chloride (1.40 g, 18.00 mmol) were added sequentially. The mixture was stirred at 0°C for 1 hour. The reaction system was diluted with dichloromethane (100 mL), and the organic phase was washed with saturated brine (25 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by slurrying with methyl tert-butyl ether (50 mL) to obtain compound 5b (2.80 g) in a yield of 87%.

[0165] MS-ESI calculated value [M+H] + 190, the actual measured value is 190.

[0166] Step 2

[0167] At -20 ° C, 5b (2.00 g, 10.60 mmol) was slowly added to concentrated sulfuric acid (35 mL) in batches, stirred and dissolved, and potassium nitrate (1.07 g, 10.58 mmol) was added in batches. The mixture was stirred and reacted at -20 ° C for 1 hour. The reaction system was slowly added dropwise to ice water (100 mL), the aqueous phase was extracted with dichloromethane (500 mL), the organic phase was dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by beating with methyl tert-butyl ether (25 mL) to obtain the first batch of compound 5c (1.00 g). After the mother liquor was concentrated, it was further beating with methyl tert-butyl ether (15 mL) to obtain the second batch of compound 5c (1.20 g). The total yield was 89%.

[0168] MS-ESI calculated value [M+H] + 235, the actual measured value is 235.

[0169] Step 3

[0170] Compound 5c (2.10 g, 8.97 mmol) was dissolved in acetone (60 mL), and an aqueous solution of magnesium sulfate (1.51 g, 12.56 mmol, 1.5 M) was added. The mixture was cooled to 0°C, and potassium permanganate (5.95 g, 37.68 mmol) was added portionwise. After complete addition, the mixture was stirred at 0°C for 1.5 hours. The reaction mixture was filtered to remove insoluble solids, and the filter cake was washed with dichloromethane (200 mL). The organic phase was washed with saturated brine (50 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to obtain compound 5d (400 mg) in a yield of 18%.

[0171] MS-ESI calculated value [M+H] + 249, the actual measured value is 249.

[0172] 1 H NMR (400MHz, DMSO-d6) δ12.28(s,1H),8.64(d,J=9.2Hz,1H),8.19(d,J=9.6Hz,1 H),3.12(t,J=6.0Hz,2H),2.75(t,J=6.4Hz,2H),2.21(s,3H),2.06–1.95(m,2H).

[0173] Step 4

[0174] 5d (400 mg, 1.61 mmol) was dissolved in aqueous hydrochloric acid (6 N, 6 mL) and stirred at 80°C for 2.5 hours. The reaction solution was diluted with water (25 mL) and the pH was adjusted to approximately 9 by slowly adding sodium bicarbonate. The aqueous phase was extracted with dichloromethane (100 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to yield the crude product 5e (350 mg).

[0175] MS-ESI calculated value [M+H] + 207, the measured value is 207.

[0176] Step 5

[0177] The crude product 5e (350 mg) from the previous step was dissolved in anhydrous dichloromethane (25 mL). Pyridine (254 mg, 3.22 mmol) and trifluoroacetic anhydride (676 mg, 3.22 mmol) were added sequentially under ice-cooling. The mixture was stirred at 0°C for 0.5 hours. The reaction system was diluted with dichloromethane (50 mL). The organic phase was washed with saturated brine (25 mL x 2), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to obtain compound 5f (410 mg). The two-step yield was 84%.

[0178] MS-ESI calculated value [M+H] + 303, the actual measured value is 303.

[0179] 1 H NMR (400MHz, CDCl3) δ13.64(s,1H),8.76(d,J=9.2Hz,1H),8.14(d,J=9.6Hz,1H),3.27(t,J=6.0Hz,2H),2.82(t,J=6.4Hz,2H),2.21–2.11(m,2H).

[0180] Step 6

[0181] 5f (410 mg, 1.36 mmol) was dissolved in a mixed solvent of methanol (50 mL), water (4 mL), and formic acid (4 mL). Zinc powder (1.74 g, 27.20 mmol) was added portionwise under ice-cooling. The mixture was stirred at 0°C for 0.5 h. The reaction system was filtered to remove insoluble matter, and the filter cake was washed sequentially with ethyl acetate (200 mL) and water (50 mL). Sodium bicarbonate was slowly added to the filtrate to adjust the pH to 8-9. The layers were separated, and the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 5 g (330 mg) of crude product.

[0182] MS-ESI calculated value [M+H] + 273, the measured value is 273.

[0183] Step 7

[0184] 5 g (330 mg) of the crude product from the previous step was dissolved in anhydrous dichloromethane (25 mL). Triethylamine (260 mg, 2.57 mmol) and acetyl chloride (200 mg, 2.57 mmol) were added sequentially under ice-cooling, and the mixture was stirred at 0°C for 1 hour. The reaction system was diluted with dichloromethane (50 mL), and the organic phase was washed with saturated brine (25 mL x 2), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 5h (310 mg). The two-step yield was 72%.

[0185] MS-ESI calculated value [M+H] + 315, the actual measured value is 315.

[0186] Step 8

[0187] 5h (310 mg, 0.99 mmol) was dissolved in a mixed solvent of methanol (22 mL) and water (1.5 mL), potassium carbonate (550 mg, 3.99 mmol) was added, and the mixture was heated to 50°C and stirred for 1 hour. The reaction system was diluted with dichloromethane (100 mL) and water (25 mL), and the liquids were separated. The aqueous phase was washed with a mixed solvent of dichloromethane / methanol (V 二氯甲烷 :V 甲醇 =4:1, 50 mL), the organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to give compound 5i (205 mg) in a yield of 95%.

[0188] MS-ESI calculated value [M+H] + 219, the actual measured value is 219.

[0189] Step 9

[0190] To a 100 mL Schlenk reaction tube were added 5i (200 mg, 0.92 mmol), 1 (257 mg, 0.92 mmol), and p-toluenesulfonic acid (158 mg, 0.92 mmol) in sequence. Anhydrous toluene (10 mL) was added, and the mixture was heated to 110°C with stirring for 6 hours. The reaction system was concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to afford crude product 5j (170 mg), which was used directly in the next step.

[0191] MS-ESI calculated value [M+H] + 462, the actual measured value is 462.

[0192] Step 10

[0193] The crude product 5j (170 mg) from the previous step was dissolved in aqueous hydrochloric acid (6N, 2 mL), and the mixture was heated to 80°C and stirred for 4 hours. The reaction system was quenched with a dichloromethane / methanol mixed solvent (V 二氯甲烷 :V 甲醇 =10:1, 100 mL) and water (20 mL), slowly add sodium bicarbonate to adjust the pH to about 9, separate the liquids, and wash the aqueous phase with a dichloromethane / methanol mixed solvent (V 二氯甲烷 :V 甲醇 =10:1, 100 mL) and the organic phases were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to give compound 5 (100 mg). The two-step yield was 26%.

[0194] MS-ESI calculated value [M+H] + 420, the actual measured value is 420.

[0195] 1 H NMR (400MHz, DMSO-d6) δ7.76(d,J=8.8Hz,1H),7.67(s,1H),7.32(d,J=9.2Hz,1H),6.62(s,1H),5.91(d,J=16.4Hz,1H),5.72(s,2H),5.49(d ,J=16.8Hz,1H),5.42(s,2H),3.09(t,J=5.6Hz,2H),2.76(t,J=6.0Hz,2H),2.07–1.98(m,2H),1.88(q,J=7.2Hz,2H),0.86(t,J=7.2Hz,3H).

[0196] Example 6

[0197] first step

[0198] 6a (20.00 g, 159.80 mmol) was dissolved in a dichloromethane / methanol mixed solvent (V 二氯甲烷 :V 甲醇 =1:1, 350mL), cooled to 0℃, dissolved liquid bromine (63.84g, 399.50mmol) in dichloromethane / methanol mixed solvent (V 二氯甲烷 :V 甲醇=1:1, 50 mL) and then added dropwise to the above solution. After the addition was complete, the mixture was stirred at 25°C for 4 hours. The reaction solution was concentrated under reduced pressure to obtain a residue, which was diluted with sodium thiosulfate aqueous solution (1M, 350 mL) and ethyl acetate (350 mL). The mixture was stirred for 10 minutes, then sodium carbonate aqueous solution (1M, 50 mL) was added and the layers separated. The organic phase was washed sequentially with sodium thiosulfate aqueous solution (1M, 230 mL x 1) and saturated brine (230 mL x 1), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to obtain compound 6b (40 g) in an 89% yield.

[0199] 1 H NMR (400MHz, CDCl3) δ7.16 (d, J = 8.4Hz, 1H), 4.40 (br s, 2H), 2.27 (d, J = 2.4Hz, 3H).

[0200] Step 2

[0201] Toluenesulfonic acid (1.83 g, 10.63 mmol) was dissolved in anhydrous acetonitrile (15 mL), stirred and dissolved, and then cooled to 0°C. 6b (1.00 g, 3.53 mmol) was added, and a solution of potassium iodide (1.47 g, 8.86 mmol) and sodium nitrite (480 mg, 7.10 mmol) in water (2 mL) was added dropwise to the reaction solution. The mixture was stirred at 0°C for 10 min, then returned to room temperature and stirred for 1 hour. The reaction system was added to water (8 mL), and the aqueous phase was adjusted to pH 9 with saturated sodium bicarbonate aqueous solution. Then, sodium thiosulfate aqueous solution (2.5 mL, 2 M) was added, and the aqueous phase was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (ethyl acetate: petroleum ether = 0-100%) to obtain compound 6c (500 mg) in a yield of 36%.

[0202] Step 3

[0203] Under a nitrogen atmosphere, 6c (9.00 g, 22.85 mmol) was dissolved in anhydrous toluene (54 mL), cooled to -30°C, and isopropylmagnesium chloride in tetrahydrofuran (2.82 g, 27.42 mmol, 2 M) was added dropwise. After addition, the reaction was maintained at -30°C with stirring for 1.5 hours. N,N-dimethylformamide (5.51 g, 75.42 mmol) was added dropwise. After complete addition, the mixture was returned to 20°C and stirred for another 2 hours. The reaction solution was diluted with saturated aqueous ammonium chloride (150 mL), and the aqueous phase was extracted with ethyl acetate (150 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to give compound 6d (4.2 g) in a yield of 62%.

[0204] 1 H NMR (400MHz, CDCl3) δ10.20 (s, 1H), 7.38 (d, J = 8.8Hz, 1H), 2.35 (d, J = 2.4Hz, 3H).

[0205] Step 4

[0206] Compound 6d (4.20 g, 14.19 mmol) was dissolved in anhydrous 1,2-dichloroethane (35 mL). Ethylene glycol (4.43 g, 71.45 mmol), triethyl orthoformate (2.31 g, 5.61 mmol), and p-toluenesulfonic acid (244.40 mg, 1.42 mmol) were added. The mixture was heated to 80°C and stirred for 12 hours. The reaction solution was washed sequentially with saturated aqueous sodium carbonate (80 mL) and saturated aqueous ammonium chloride (80 mL). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to obtain compound 6e (3.9 g) in an 81% yield.

[0207] 1 H NMR (400MHz, CDCl3) δ7.33 (d, J = 8.8Hz, 1H), 6.43 (s, 1H), 4.36-4.30 (m, 2H), 4.11-4.04 (m, 2H), 2.32 (d, J = 1.2Hz, 3H).

[0208] Step 5

[0209] Under nitrogen, 6e (1.5 g, 4.41 mmol), benzophenone imine (880 mg, 4.85 mmol), palladium acetate (99 mg, 0.44 mmol), (255 mg, 0.44 mmol), and sodium tert-butoxide (850 mg, 8.82 mmol) were dissolved in anhydrous toluene (15 mL). The mixture was heated to 100°C and stirred for 12 hours. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to obtain compound 6f (300 mg) in a yield of 15%.

[0210] 1 H NMR (400MHz, CDCl3) δ7.76 (d, J = 7.2Hz, 2H), 7.52-7.44 (m, 1H), 7.44-7.36 (m, 2H), 7.34-7.27 (m, 3H), 7.26-7 .21(m,2H),6.47(s,1H),5.91(d,J=10.4Hz,2H),4.10-4.04(m,2H),3.96-3.90(m,2H),2.21(d,J=2.0Hz,3H).

[0211] Step 6

[0212] Under nitrogen protection, (but-3-en-1-yloxy)(tert-butyl)dimethylsilane (30 mg, 0.16 mmol) was dissolved in anhydrous toluene (1 mL), cooled to 0°C, and 9-borabicyclo[3.3.1]nonane (24 mg, 0.19 mmol) was added. The mixture was heated to 80°C and stirred for 20 min, then cooled to 0°C. A solution of sodium hydroxide (13 mg, 0.32 mmol) in water (1 mL) was added dropwise. The mixture was stirred at 0°C for 10 min, and 6f (57 mg, 0.13 mmol), tetrabutylammonium iodide (3 mg, 0.008 mmol) and 1,1'-bis(diphenylphosphino)ferrocenepalladium chloride (3 mg, 0.003 mmol) were added. After the addition, the mixture was heated to 80°C and stirred for 12 hours. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by preparative thin-layer chromatography to obtain compound 6g (40 mg). Yield: 56%.

[0213] Step 7

[0214] 6 g (120 mg, 0.22 mmol) was dissolved in anhydrous ethanol (4 mL), and compound 1 (62 mg, 0.22 mmol) and concentrated hydrochloric acid (52.56 mg, 1.44 mmol, 12 N) were added. The mixture was heated to 80°C and stirred for 2 hours. The reaction solution was concentrated to obtain a residue, which was purified by preparative thin-layer chromatography to obtain compound 6 (7 mg), with a yield of 7%.

[0215] MS-ESI calculated value [M+H] + 469, the actual measured value is 469.

[0216] 1 H NMR (500MHz, DMSO-d6) δ8.98(s,1H),7.80(d,J=10.5Hz,1H),7.79(s,1H),5.92(d,J=16.5Hz,1H),5.50(d,J=16.5Hz,1H ),5.497(s,2H),3.19-3.12(m,2H),2.43(d,J=2.0Hz,3H),1.92-1.84(m,2H),1.68-1.55(m,4H),0.86(t,J=7.0Hz,3H).

[0217] Example 7

[0218] first step

[0219] 7a (11.00 g, 67.00 mmol) was dissolved in trifluoroacetic acid (165 mL), and sodium nitrite (13.78 g, 199.72 mmol) was added portionwise. The mixture was stirred at 25°C for 12 hours. The reaction system was diluted with water (100 mL) and extracted with dichloromethane (100 mL x 3). The organic phases were combined and washed with saturated aqueous sodium bicarbonate (100 mL x 3), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to obtain compound 7b (7.20 g) in a yield of 51%.

[0220] MS-ESI calculated value [M+H] + 210, the actual measured value is 210.

[0221] Step 2

[0222] 7b (7.20 g, 34.42 mmol) was dissolved in a tetrahydrofuran / water mixed solvent (V 四氢呋喃 :V 水 =3:1, 35 mL), wet palladium on carbon (700 mg, 6.58 mmol, 10% w / w) was added, and the mixture was stirred at room temperature under a hydrogen atmosphere for 12 hours. The reaction system was filtered to remove insoluble matter, the filter cake was rinsed with dichloromethane, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to obtain compound 7c (5.60 g) in a yield of 91%.

[0223] MS-ESI calculated value [M+H] + 180, the actual measured value is 180.

[0224] 1 H NMR (400MHz, DMSO-d6) δ7.16(s,1H),6.30(s,1H),5.92(s,2H),2.38(s,3H).

[0225] Step 3

[0226] 7c (5.50 g, 30.70 mmol) was dissolved in dichloromethane (110 mL). The mixture was cooled to 0°C, and N,N-diisopropylethylamine (5.95 g, 46.04 mmol) and acetyl chloride (3.61 g, 46.04 mmol) were added dropwise. After complete addition, the reaction was stirred at room temperature for 1 hour. The reaction solution was quenched with methanol (5 mL) and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to obtain compound 7d (6.60 g) in a yield of 96%.

[0227] MS-ESI calculated value [M+H] + 222, the actual measured value is 222.

[0228] 1 H NMR (400MHz, CDCl3) δ12.06(br s,1H),8.35(s,1H),7.24(s,1H),6.01(s,2H),2.55(s,3H),2.19(s,3H).

[0229] Step 4

[0230] 7d (6.60 g, 29.84 mmol) was dissolved in glacial acetic acid (105 mL). The mixture was cooled to 0°C, and liquid bromine (4.77 g, 29.84 mmol) and hydrobromic acid acetic acid solution (12 M, 4.83 g, 59.68 mmol) were added dropwise. After the addition was complete, the reaction was stirred at room temperature for 1 hour. The reaction solution was poured into ice water (100 mL). The precipitated solid was filtered, and the filter cake was dissolved in dichloromethane (100 mL). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to give compound 7e (7.00 g) in a yield of 78%.

[0231] MS-ESI calculated value [M+H] + 300,302, the measured value is 300,302.

[0232] Step 5

[0233] 7e (5.90 g, 19.67 mmol) was dissolved in anhydrous ethanol (60 mL), followed by the addition of concentrated hydrochloric acid (12 M, 5.0 mL, 60.00 mmol). After complete addition, the mixture was heated to 80°C and stirred for 4 hours. The reaction solution was concentrated under reduced pressure to obtain a residue, which was then purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to afford crude product 7f (4.56 g).

[0234] MS-ESI calculated value [M+H] + 214,216, the measured value is 214,216.

[0235] Step 6

[0236] 7f (4.0 g, 18.70 mmol) was dissolved in anhydrous dichloromethane (40 mL), followed by the addition of benzylamine (10 mL). After complete addition, the mixture was stirred at room temperature for 4 hours. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to afford compound 7g (2.5 g), with a two-step yield of 47%.

[0237] MS-ESI calculated value [M+H] + 285, the actual measured value is 285.

[0238] Step 7

[0239] Dissolve 7g (2.0g, 7.04mmol) in anhydrous tetrahydrofuran (25mL), then add wet palladium on carbon (200mg, 10% w / w). Stir the mixture at 50°C under a hydrogen atmosphere for 20 hours. The reaction mixture is filtered to remove insoluble matter, and the filtrate is concentrated under reduced pressure to obtain the crude product 7h (1.3g).

[0240] MS-ESI calculated value [M+H] + 195, the actual measured value is 195.

[0241] Step 8

[0242] The crude product 7h (1.3 g) from the previous step was dissolved in a mixed solvent of tetrahydrofuran / water (V 四氢呋喃 :V 水=2:1, 24 mL), potassium carbonate (1.85 g, 13.40 mmol) and 9-fluorenylmethyl chloroformate (1.38 g, 5.36 mmol) were added, and the reaction system was stirred at 0°C for 1 hour. The reaction system was diluted with water (20 mL) and ethyl acetate (100 mL). The layers were separated, and the organic phase was washed with saturated aqueous sodium chloride (250 mL x 2), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to obtain compound 7i (820 mg), with a two-step yield of 28%.

[0243] MS-ESI calculated value [M+H] + 417, the actual measured value is 417.

[0244] Step 9

[0245] To a 100 mL Schlenk reaction tube were added 7i (200 mg, 0.48 mmol), 1 (107 mg, 0.38 mmol), and p-toluenesulfonic acid (34 mg, 0.20 mmol) in sequence. Anhydrous toluene (10 mL) was added, and the mixture was heated to 110°C with stirring for 6 hours. The reaction system was concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to afford crude product 7j (200 mg), which was used directly in the next step.

[0246] MS-ESI calculated value [M+H] + 660, the actual measured value is 660.

[0247] Step 10

[0248] Dissolve 7j (200 mg, 0.30 mmol) in anhydrous tetrahydrofuran (5 mL), add piperidine (255 mg, 3.00 mmol), and stir at room temperature for 3.5 hours. The reaction system is concentrated under reduced pressure to obtain a crude product, which is purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to afford compound 7 (80 mg) in a 61% yield.

[0249] MS-ESI calculated value [M+H] + 438, the actual measured value is 438.

[0250] 1H NMR(500MHz,DMSO-d6)δ7.72(s,1H),7.66(s,1H),7.51(s,1H),6.29(d,J=2.0Hz,2H),5.91(d, J=16.5Hz,1H),5.48(d,J=16.5Hz,1H),4.28(s,2H),1.92-1.84(m,2H),0.86(t,J=7.0Hz,3H).

[0251] Example 8

[0252] first step

[0253] To a single-necked flask (25 mL) were added 7 (30 mg, 0.07 mmol) and glycolic acid (10.6 mg, 0.14 mmol) in sequence, followed by anhydrous dichloromethane (1.0 mL). The reaction system was cooled to 0°C, and N,N-diisopropylethylamine (18 mg, 0.14 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (24 mg, 0.06 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 8 (12 mg) in a yield of 35%.

[0254] MS-ESI calculated value [M+H] + 496, the actual measured value is 496.

[0255] Example 9

[0256] first step

[0257] To a single-necked flask (25 mL) were added 7 (65 mg, 0.15 mmol) and N-tert-butyloxycarbonyl-glycine (52 mg, 0.30 mmol) in sequence, followed by anhydrous dichloromethane (2 mL). The reaction system was cooled to 0°C, and N,N-diisopropylethylamine (39 mg, 0.38 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (54 mg, 0.14 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 9a (80 mg) in a 90% yield.

[0258] MS-ESI calculated value [M+H] + 595, the actual measured value is 595.

[0259] Step 2

[0260] 9a (60 mg, 0.10 mmol) was dissolved in anhydrous dichloromethane (4 mL). Trifluoroacetic acid (0.4 mL) was added under ice-cooling. The reaction system was returned to room temperature and stirred for 2.5 hours. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 9 (15 mg) in a 30% yield.

[0261] MS-ESI calculated value [M+H] + 495, the actual measured value is 495.

[0262] 1 H NMR (500MHz, DMSO-d6) δ7.74(s,2H),7.58(s,1H),6.32(d,J=2.0Hz,2H),5.92(d,J=16.5Hz,1H),5.7 3(s,2H),5.50(d,J=16.5Hz,1H),4.85(s,2H),3.56(s,2H),1.92-1.84(m,2H),0.85(t,J=7.0Hz,3H).

[0263] Example 10

[0264] first step

[0265] 7b (500 mg, 2.39 mmol) was dissolved in ethanol (2 mL), and paraformaldehyde (1.08 g, 35.64 mmol) and isopropylamine (3.43 g, 35.89 mmol) were added. The mixture was stirred at 100°C for 72 hours. The mixture was concentrated under reduced pressure to obtain a residue, which was diluted with water (10 mL). The aqueous phase was adjusted to pH 7 with saturated aqueous sodium bicarbonate solution and extracted with dichloromethane (30 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 10a (80 mg) in a yield of 12%.

[0266] MS-ESI calculated value [M+H] + 281, the measured value is 281.

[0267] 1 H NMR (400MHz, CDCl3) δ7.73 (s, 1H), 7.29 (s, 1H), 6.30 (s, 2H), 3.40-3.20 (m, 5H), 1.24 (d, J = 6.8Hz, 6H).

[0268] Step 2

[0269] 10a (80 mg, 0.29 mmol) was dissolved in a tetrahydrofuran / water mixed solvent (V 四氢呋喃 :V 水 =3:1, 2 mL), fluorenylmethyloxycarbonyl chloride (74 mg, 0.29 mmol) and sodium bicarbonate (24 mg, 0.29 mmol) were added sequentially, and the mixture was stirred at 25°C for 12 hours. The mixture was concentrated under reduced pressure to obtain a residue, which was diluted with water (10 mL). The aqueous phase was extracted with dichloromethane (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to obtain compound 10b (70 mg) in a yield of 78%.

[0270] MS-ESI calculated value [M+H] + 503, the actual measured value is 503.

[0271] Step 3

[0272] 10b (540 mg, 1.07 mmol) was dissolved in ethanol (10 mL), and wet palladium on carbon (10% w / w, 54 mg) was added. The reaction system was heated to 40°C and stirred for 4 hours. The mixture was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to obtain compound 10c (360 mg) in a yield of 71%.

[0273] MS-ESI calculated value [M+H] + 473, the measured value is 473.

[0274] Step 4

[0275] Compound 10c (150 mg, 0.32 mmol) was dissolved in anhydrous toluene (4 mL), followed by the addition of 1 (89 mg, 0.32 mmol) and p-toluenesulfonic acid (28 mg, 0.16 mmol). After complete addition, the mixture was heated to 120°C and stirred for 4 hours. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to afford compound 10d (70 mg) in a 31% yield.

[0276] MS-ESI calculated value [M+H] + 716, the actual measured value is 716.

[0277] Step 5

[0278] 10d (80 mg, 0.11 mmol) was dissolved in tetrahydrofuran (2 mL), and piperidine (19 mg, 0.22 mmol) was added. The mixture was heated to 40°C and stirred for 12 hours. The mixture was concentrated under reduced pressure to obtain a residue, which was purified by preparative thin-layer chromatography to afford compound 10 (30 mg) in a 55% yield.

[0279] MS-ESI calculated value [M+H] + 494, the actual measured value is 494.

[0280] 1 H NMR (400MHz, DMSO-d6) δ7.71(s,1H),7.63(s,1H),7.49(s,1H),6.28(s,2H),5.91(d,J=16.4Hz,1H),5.47(d,J=16.8Hz,1H),5.45-5. 40(m,1H),3.36-3.28(m,2H),3.19-3.10(m,1H),3.09-3.00(m,2H),1.93-1.82(m,2H),1.14(d,J=6.4Hz,6H),0.85(t,J=7.2Hz,3H).

[0281] Example 11

[0282] first step

[0283] Compound 11a (2.5 g, 215.14 mmol) was dissolved in ethanol (30 mL), and paraformaldehyde (6.82 g, 227.11 mmol) and isopropylamine (21.70 g, 227.06 mmol) were added. The mixture was stirred at 100°C for 72 hours. The mixture was concentrated under reduced pressure to obtain a residue, which was diluted with water (30 mL). The aqueous phase was adjusted to pH 7 with saturated aqueous sodium bicarbonate solution and extracted with dichloromethane (100 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 11b (2.20 g) in a yield of 61%.

[0284] MS-ESI calculated value [M+H] + 237, the actual measured value is 237.

[0285] Step 2

[0286] 11b (2.20 g, 9.65 mmol) was dissolved in a tetrahydrofuran / water mixed solvent (V 四氢呋喃 :V 水=3:1, 13 mL) was added with fluorenylmethyloxycarbonyl chloride (3.75 g, 14.50 mmol) and sodium bicarbonate (1.26 g, 14.50 mmol), and the mixture was stirred at 25°C for 12 hours. The mixture was concentrated under reduced pressure to obtain a residue, which was diluted with water (30 mL). The aqueous phase was extracted with dichloromethane (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to obtain compound 11c (345 mg) in a yield of 8%.

[0287] MS-ESI calculated value [M+H] + 459, the actual measured value is 459.

[0288] Step 3

[0289] 11c (345 mg, 0.72 mmol) was dissolved in ethanol (10 mL), and wet palladium on carbon (10% w / w, 35 mg) was added. The mixture was heated to 40°C and stirred for 4 hours. The mixture was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to obtain compound 11d (220 mg) in a yield of 71%.

[0290] MS-ESI calculated value [M+H] + 429, the actual measured value is 429.

[0291] Step 4

[0292] Compound 11d (100 mg, 0.23 mmol) was dissolved in anhydrous toluene (4 mL), followed by the addition of 1 (66 mg, 0.24 mmol) and p-toluenesulfonic acid (20 mg, 0.12 mmol). After complete addition, the mixture was heated to 120°C and stirred for 4 hours. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to afford compound 11e (100 mg) in a 64% yield.

[0293] MS-ESI calculated value [M+H] + 672, the actual measured value is 672.

[0294] Step 5

[0295] 11e (80 mg, 0.12 mmol) was dissolved in tetrahydrofuran (2 mL), and piperidine (21 mg, 0.25 mmol) was added. The mixture was heated to 40°C and stirred for 12 hours. The mixture was concentrated under reduced pressure to obtain a residue, which was purified by thin-layer chromatography to obtain compound 11 (6 mg) in a yield of 11%.

[0296] MS-ESI calculated value [M+H] + 450, the actual measured value is 450.

[0297] Example 12

[0298] first step

[0299] 12a (5.00 g, 22.00 mmol) was dissolved in dichloromethane (450 mL), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (12.55 g, 33.00 mmol), N,N-diisopropylethylamine (5.69 g, 44.00 mmol), and N-methyl-N-methoxyamine hydrochloride (1.41 g, 33.00 mmol) were added, and the mixture was stirred at 25 ° C for 12 hours. The reaction system was diluted with dichloromethane (200 mL), and aqueous hydrochloric acid (1N, 50 mL) was added. The layers were separated, and the organic phase was washed sequentially with saturated aqueous sodium bicarbonate (50 mL x 1) and saturated brine (50 mL x 1). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to give compound 12b (5.30 g) in a yield of 89%.

[0300] Step 2

[0301] 4-Bromo-1,2-methylenedioxybenzene (9.37 g, 46.61 mmol) was dissolved in tetrahydrofuran (100 mL). After cooling to -65°C, n-butyllithium (1.6 M in n-hexane, 29 mL) was added dropwise. The mixture was stirred at -65°C for 0.5 hours. 12b (5.04 g, 18.64 mmol) was dissolved in anhydrous tetrahydrofuran (40 mL) and added dropwise to the reaction system. After the addition was complete, the mixture was stirred at -65°C for 3 hours. The mixture was quenched with saturated aqueous ammonium chloride (500 mL) and extracted with ethyl acetate (500 mL x 3). The organic phase was washed with saturated brine (500 mL x 1). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to give compound 12c (4.00 g). Yield: 45%.

[0302] 1H NMR (400MHz, CDCl3) δ7.63(d,J=8.0Hz,1H),7.44(s,1H),6.82(d,J=8.4Hz,1H),6.03(s,2H),2.50(s,6H),1.46(s,9H).

[0303] Step 3

[0304] Reagent 12c (3.00 g, 9.05 mmol) was dissolved in dichloromethane (100 mL), cooled to 0°C, and trifluoroacetic acid (27.61 g, 242.15 mmol) was added. The mixture was heated to 25°C and stirred for 3 hours. The mixture was concentrated under reduced pressure, and the residue was dissolved in dichloromethane (40 mL). The temperature was lowered to 0°C, and N,N-diisopropylethylamine (7.42 g, 57.41 mmol) and trifluoroacetic anhydride (3.85 g, 18.33 mmol) were added dropwise. After the addition was complete, the mixture was heated to 25°C and stirred for 4 hours. The mixture was quenched with water (200 mL) and extracted with ethyl acetate (200 mL x 3). The organic phase was washed with saturated brine (200 mL x 1). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to give compound 12d (2.50 g). Yield: 84%.

[0305] Step 4

[0306] 12d (2.50 g, 7.64 mmol) was dissolved in acetic anhydride (5.8 mL), cooled to 0°C, and copper nitrate (860 mg, 4.59 mmol) was added portionwise. After complete addition, the mixture was stirred at 0°C for 3 hours. The mixture was quenched with water (30 mL) and extracted with ethyl acetate (30 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to obtain compound 12e (900 mg) in a yield of 75%.

[0307] MS-ESI calculated value [M+H] + 373, the actual measured value is 373.

[0308] 1 H NMR (400MHz, CDCl3) δ7.60(s,1H),6.66(br s,1H),6.64(s,1H),6.21(s,2H),2.43(s,6H).

[0309] Step 5

[0310] 12e (400 mg, 1.07 mmol) was dissolved in ethanol (10 mL), and wet palladium on carbon (10% w / w, 40 mg) was added. The mixture was heated to 40°C and stirred for 4 hours. The mixture was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to provide compound 12f (220 mg) in a 60% yield.

[0311] MS-ESI calculated value [M+H] + 343, the actual measured value is 343.

[0312] Step 6

[0313] 12f (100 mg, 0.29 mmol) was dissolved in anhydrous toluene (4 mL), followed by the addition of 1 (82 mg, 0.32 mmol) and p-toluenesulfonic acid (26 mg, 0.15 mmol). After complete addition, the mixture was heated to 120°C and stirred for 4 hours. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to afford compound 12g (80 mg) in a 47% yield.

[0314] Step 7

[0315] Dissolve 12 g (80 mg, 0.14 mmol) of the compound in methanol (4 mL), then add hydrochloric acid (1.6 mL, 4 M). The mixture is heated to 65°C and stirred for 4 hours. The mixture is concentrated under reduced pressure to obtain a residue, which is then purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to provide compound 12 (14 mg) in a 21% yield.

[0316] MS-ESI calculated value [M+H] + 490, the actual measured value is 490.

[0317] 1 H NMR (400MHz, DMSO-d6) δ7.74(s,1H),7.64(s,1H),7.58(s,1H),6.32(s,2H),5.92(d,J=16.4Hz, 1H),5.61(s,2H),5.50(d,J=16.4Hz,1H),2.81(s,6H),1.93-1.84(m,2H),0.84(t,J=7.2Hz,3H).

[0318] Example 13

[0319] first step

[0320] 13a (15 g, 73.90 mmol) was added portionwise to a mixture of concentrated sulfuric acid (45 mL) and concentrated nitric acid (12 mL) under an ice bath. After complete addition, the reaction system was gradually returned to room temperature and stirred for 5 hours. The reaction solution was poured into ice water (1 L), whereupon a yellow solid precipitated. This was filtered and dried to afford compound 13b (22 g).

[0321] Step 2

[0322] 13b (750 mg, 3.02 mmol), tert-butyl carbamate (424 mg, 3.62 mmol), Pd2(dba)3 (138 mg, 0.15 mmol), XPhos (288 mg, 0.60 mmol), and cesium carbonate (1.96 g, 6.04 mmol) were added to a Schlenk reaction tube. The atmosphere was purged with nitrogen three times, and then anhydrous toluene (8 mL) was added. The tube was sealed and stirred in a 90°C oil bath overnight. The reaction solution was diluted with ethyl acetate (100 mL), and the organic phase was washed sequentially with water (25 mL x 1) and brine (25 mL x 2), dried, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to obtain compound 13c (550 mg) in a yield of 64%.

[0323] 1 H NMR (400MHz, DMSO-d6) δ10.44(s,1H),8.75(d,J=6.0Hz,1H),7.92(d,J=8.4Hz,1H),7.05(s,1H),1.55(s,9H).

[0324] Step 3

[0325] 13c (500 mg, 1.76 mmol) and ammonium chloride (156 mg, 2.94 mmol) were dissolved in a mixture of ethanol (8 mL) and water (2 mL). Iron powder (394 mg, 7.04 mmol) was added portionwise. The reaction system was heated to 60°C and stirred for 2 hours. The reaction solution was diluted with ethyl acetate (100 mL). The organic phase was washed sequentially with water (25 mL x 1) and brine (25 mL x 1), dried, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to obtain compound 13d (310 mg) in a yield of 69%.

[0326] MS-ESI calculated value [M+H] + 255, the measured value is 255.

[0327] Step 4

[0328] To a 100 mL Schlenk reaction tube, 13d (250 mg, 0.98 mmol), 1 (167 mg, 0.60 mmol), and p-toluenesulfonic acid (26 mg, 0.15 mmol) were added sequentially. Anhydrous toluene (3 mL) was added, and the mixture was heated to 110°C and stirred for 2 hours. The reaction system was concentrated under reduced pressure to obtain the crude product, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to afford crude product 13e (180 mg), which was used directly in the next step.

[0329] MS-ESI calculated value [M+H] + 498, the actual measured value is 498.

[0330] Step 5

[0331] Compound 13e (45 mg, 0.09 mmol) was dissolved in anhydrous dichloromethane (1 mL). Trifluoroacetic acid (0.3 mL) was added under ice-cooling. The reaction system was returned to room temperature and stirred for 5 hours. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 9 (20 mg) in a 30% yield.

[0332] MS-ESI calculated value [M+H] + 398, the actual measured value is 398.

[0333] 1 H NMR (400MHz, DMSO-d6) δ8.39(s,1H),7.80(d,J=12.4Hz,1H),7.70(s,1H),7.16(d,J=9.6Hz,1H),5. 90(d,J=16.4Hz,1H),5.46(d,J=16.4Hz,1H),5.43(s,2H),1.93-1.80(m,2H),0.85(t,J=7.2Hz,3H).

[0334] Example 14

[0335] first step

[0336] To a single-necked flask (25 mL) were added 7 (20 mg, 0.05 mmol) and 2-cyclopropyl-2-hydroxyacetic acid (12 mg, 0.10 mmol) in sequence, followed by anhydrous dichloromethane (1.0 mL). The reaction system was cooled to 0°C, and triethylamine (10 mg, 0.10 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (23 mg, 0.06 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 14 (12 mg) in a yield of 49%.

[0337] MS-ESI calculated value [M+H] + 536, the actual measured value is 536.

[0338] 1 H NMR(400MHz,DMSO-d6)δ8.66(t,J=5.6Hz,1H),7.87(s,1H),7.74(s,1H),7. 54(s,1H),6.30(s,2H),5.92(d,J=16.4Hz,1H),5.74(s,2H),5.49(d,J=16. 4Hz,1H),4.75(d,J=5.2Hz,2H),3.56-3.51(m,1H),1.94-1.83(m,2H),1.07 -0.97(m,1H),0.85(t,J=7.2Hz,3H),0.39-0.29(m,2H),0.28-0.18(m,2H).

[0339] Example 15

[0340] first step

[0341] To a single-necked flask (25 mL) were added 7 (20 mg, 0.05 mmol) and lactic acid (9 mg, 0.10 mmol) in sequence, followed by anhydrous dichloromethane (1.0 mL). The reaction system was cooled to 0°C, and N,N-diisopropylethylamine (20 mg, 0.15 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (20 mg, 0.05 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 15 (10 mg) in a yield of 43%.

[0342] MS-ESI calculated value [M+H]+ 510, the actual measured value is 510.

[0343] 1 H NMR (400MHz, DMSO-d6) δ8.74(t,J=5.2Hz,1H),7.89(s,1H),7.76(s,1H),7.57(s,1H),6.32(s,2H),5.94(d,J=16.8Hz,1H) ,5.74(s,2H),5.51(d,J=16.4Hz,1H),4.77(d,J=5.6Hz,2H),4.05-3.96(m,1H),1.94-1.82(m,2H),0.88(t,J=7.2Hz,3H).

[0344] Example 16

[0345] first step

[0346] To a single-necked flask (25 mL) were added 7 (20 mg, 0.05 mmol) and 3-hydroxycyclobutanecarboxylic acid (12 mg, 0.10 mmol) in sequence, followed by anhydrous dichloromethane (1.0 mL). The reaction system was cooled to 0°C, and N,N-diisopropylethylamine (17 mg, 0.13 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (23 mg, 0.06 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 16 (15 mg) in a yield of 61%.

[0347] MS-ESI calculated value [M+H] + 536, the actual measured value is 536.

[0348] 1 H NMR (400MHz, DMSO-d6) δ7.72(s,1H),7.69(s,1H),7.52(s,1H),6.29(s,2H),5.91(d,J=16.4Hz,1H),5.58(s,2H),5.48(d,J=16. 8Hz,1H),4.77-4.69(m,2H),3.97-3.82(m,1H),2.45-2.35(m,1H),2.30-2.22(m,2H),2.00-1.82(m,4H),0.85(t,J=7.2Hz,3H).

[0349] Example 17

[0350] first step

[0351] To a single-necked flask (25 mL) were added 7 (20 mg, 0.05 mmol) and 4-hydroxycyclohexanecarboxylic acid (14 mg, 0.10 mmol) in sequence, followed by anhydrous dichloromethane (1.0 mL). The reaction system was cooled to 0°C, and N,N-diisopropylethylamine (17 mg, 0.13 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (23 mg, 0.06 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 17 (9 mg) in a yield of 35%.

[0352] MS-ESI calculated value [M+H] + 564, the actual measured value is 564.

[0353] 1 H NMR(400MHz,DMSO-d6)δ8.60(s,1H),7.73(s,1H),7.71(s,1H),7.54(s,1H),6.30( s,2H),5.92(d,J=16.8Hz,1H),5.67-5.57(m,2H),5.48(d,J=16.4Hz,1H),4.79-4.7 0(m,2H),3.35-3.22(m,1H),2.12-2.00(m,1H),1.92-1.84(m,2H),1.84-1.77(m,2 H),1.77-1.67(m,2H),1.42-1.32(m,2H),1.15-1.02(m,2H),0.85(t,J=7.2Hz,3H).

[0354] Example 18

[0355] first step

[0356] To a single-necked flask (25 mL) were added 7 (20 mg, 0.05 mmol) and (R)-3-hydroxybutyric acid (10 mg, 0.10 mmol) in sequence, followed by anhydrous dichloromethane (1.0 mL). The reaction system was cooled to 0°C, and N,N-diisopropylethylamine (20 mg, 0.15 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (23 mg, 0.06 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 18 (17 mg) in a yield of 71%.

[0357] MS-ESI calculated value [M+H] + 524, the actual measured value is 524.

[0358] 1 H NMR (400MHz, DMSO-d6) δ7.730(s,1H),7.726(s,1H),7.53(s,1H),6.29(d,J=2.0Hz,2H),5.92(d,J=16.4Hz,1H),5.64(s,2H),5.49(d,J=16.4Hz, 1H),4.81-4.65(m,2H),4.00-3.89(m,1H),2.29-2.20(m,1H),2.17-2.09 (m,1H),1.94-1.82(m,2H),0.99(d,J=6.4Hz,3H),0.85(t,J=7.2Hz,3H).

[0359] Example 19

[0360] first step

[0361] To a single-necked flask (25 mL) were added 7 (20 mg, 0.05 mmol) and (S)-3-hydroxybutyric acid (10 mg, 0.10 mmol) in sequence, followed by anhydrous dichloromethane (1.0 mL). The reaction system was cooled to 0°C, and N,N-diisopropylethylamine (20 mg, 0.15 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (23 mg, 0.06 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 19 (15 mg) in a yield of 63%.

[0362] MS-ESI calculated value [M+H] +524, the actual measured value is 524.

[0363] 1 H NMR (400MHz, DMSO-d6) δ8.66(t,J=5.6Hz,1H),7.73(s,2H),7.54(s,1H),6.30(s,2H),5.92(d,J=16.4Hz,1H),5.68-5.62(m,2H),5.49(d,J=16.4Hz ,1H),4.77-4.71(m,2H),4.00-3.89(m,1H),2.28-2.21(m,1H),2.17-2.1 0(m,1H),1.93-1.84(m,2H),1.00(d,J=6.4Hz,3H),0.85(t,J=7.2Hz,3H).

[0364] Example 20

[0365] first step

[0366] To a single-necked flask (25 mL) were added 2 (20 mg, 0.05 mmol) and 3-hydroxycyclobutanecarboxylic acid (12 mg, 0.10 mmol) in sequence, followed by anhydrous dichloromethane (1.0 mL). The reaction system was cooled to 0°C, and N,N-diisopropylethylamine (20 mg, 0.15 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (23 mg, 0.06 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 20 (15 mg) in a yield of 61%.

[0367] MS-ESI calculated value [M+H] + 524, the actual measured value is 524.

[0368] 1 H NMR (400MHz, DMSO-d6) δ8.30(d,J=8.0Hz,1H),7.89(d,J=10.8Hz,1H),7.79(s,1H),5.91(d,J=16.4Hz,1H),5.61(s,2H),5.49(d,J= 16.8Hz,1H),4.89-4.81(m,2H),3.98-3.85(m,1H),2.46-2.38(m,1H),2.45-2.35(m,2H),1.98-1.82(m,4H),0.85(t,J=7.2Hz,3H).

[0369] Example 21

[0370] first step

[0371] To a single-necked flask (25 mL) were added 2 (20 mg, 0.05 mmol) and 4-hydroxycyclohexanecarboxylic acid (14 mg, 0.10 mmol) in sequence, followed by anhydrous dichloromethane (1.0 mL). The reaction system was cooled to 0°C, and N,N-diisopropylethylamine (20 mg, 0.15 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (23 mg, 0.06 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 20 (22 mg) in a yield of 89%.

[0372] MS-ESI calculated value [M+H] + 552, the actual measured value is 552.

[0373] 1 H NMR (400MHz, DMSO-d6) δ8.30(d,J=8.0Hz,1H),7.91(d,J=10.4Hz,1H),7.79(s,1H),5.92(d,J=16.8Hz,1H),5.66-5.60(m,2H),5.49(d,J=16.4H z,1H),3.36-3.25(m,1H),2.14-2.04(m,1H),1.92-1.85(m,2H),1.85-1 .68(m,4H),1.55-1.35(m,2H),1.16-1.02(m,2H),0.86(t,J=7.2Hz,3H).

[0374] Example 22

[0375] first step

[0376] To a single-necked flask (25 mL) were added 2 (25 mg, 0.06 mmol) and lactic acid (11 mg, 0.10 mmol) in sequence, followed by anhydrous dichloromethane (1.0 mL). The reaction system was cooled to 0°C, and triethylamine (23 mg, 0.18 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (27 mg, 0.07 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 22 (28 mg) in a yield of 96%.

[0377] MS-ESI calculated value [M+H] + 498, the actual measured value is 498.

[0378] 1 H NMR (400MHz, DMSO-d6) δ8.74(t,J=6.0Hz,1H),8.42(d,J=8.0Hz,1H),7.92(d,J=10.8Hz,1H),7.79(s,1H),5.92(d,J=16.4Hz,1 H),5.78-5.68(m,2H),5.50(d,J=16.4Hz,1H),5.00-4.76(m,2H),4.06-3.95(m,1H),1.94-1.82(m,2H),0.86(t,J=7.2Hz,3H).

[0379] Example 23

[0380] first step

[0381] To a single-necked flask (25 mL) were added 2 (25 mg, 0.06 mmol) and 2-cyclopropyl-2-hydroxyacetic acid (14 mg, 0.12 mmol) in sequence, followed by anhydrous dichloromethane (1.0 mL). The reaction system was cooled to 0°C, and N,N-diisopropylethylamine (23 mg, 0.18 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (34 mg, 0.09 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 23 (28 mg) in a yield of 91%.

[0382] MS-ESI calculated value [M+H] + 524, the actual measured value is 524.

[0383] 1H NMR (400MHz, DMSO-d6) δ8.69(t,J=5.6Hz,1H),8.43(d,J=7.6Hz,1H),7.92(d, J=10.8Hz,1H),7.80(s,1H),5.92(d,J=16.4Hz,1H),5.78-5.70(m,2H),5.50( d,J=16.4Hz,1H),5.00-4.79(m,2H),3.57-3.52(m,1H),1.94-1.82(m,2H),1. 09-0.97(m,1H),0.86(t,J=6.8Hz,3H),0.38-0.28(m,2H),0.28-0.18(m,2H).

[0384] Example 24

[0385] first step

[0386] To a single-necked flask (25 mL) were added 2 (20 mg, 0.05 mmol) and (S)-3-hydroxybutyric acid (10 mg, 0.10 mmol) in sequence, followed by anhydrous dichloromethane (1.0 mL). The reaction system was cooled to 0°C, and N,N-diisopropylethylamine (19 mg, 0.15 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (23 mg, 0.06 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 24 (18 mg) in a yield of 75%.

[0387] MS-ESI calculated value [M+H] + 512, the actual measured value is 512.

[0388] 1 H NMR (400MHz, DMSO-d6) δ8.32(d,J=8.4Hz,1H),7.91(d,J=10.4Hz,1H),7.79(s,1H),5.92(d,J=16.8Hz,1H),5.66(s,2H),5.50(d,J=16.4Hz,1H ),4.91-4.76(m,2H),4.00-3.90(m,1H),2.31-2.22(m,1H),2.20-2.10( m,1H),1.93-1.84(m.2H),0.99(d,J=6.4Hz,3H),0.85(d,J=7.2Hz,3H).

[0389] Example 25

[0390] first step

[0391] To a single-necked flask (25 mL) were added 2 (25 mg, 0.06 mmol) and (R)-3-hydroxybutyric acid (13 mg, 0.12 mmol) in sequence, followed by anhydrous dichloromethane (1.0 mL). The reaction system was cooled to 0°C, and N,N-diisopropylethylamine (23 mg, 0.18 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (27 mg, 0.07 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 25 (18 mg) in a yield of 60%.

[0392] MS-ESI calculated value [M+H] + 512, the actual measured value is 512.

[0393] 1 H NMR (400MHz, DMSO-d6) δ8.30(d,J=8.4Hz,1H),7.89(d,J=10.8Hz,1H),7.78(s,1H),5.91(d,J=16.4Hz,1H),5.72-5.56(m,2H),5.49(d,J=16.8Hz, 1H),4.92-4.76(m,2H),4.00-3.90(m,1H),2.30-2.22(m,1H),2.20-2.10 (m,1H),1.94-1.82(m.2H),0.99(d,J=6.4Hz,3H),0.85(d,J=7.2Hz,3H).

[0394] Example 26

[0395] first step

[0396] To a single-necked flask (25 mL) were added 26a (30 mg, 0.06 mmol) and (S)-3-hydroxybutyric acid (10 mg, 0.12 mmol) in sequence, followed by anhydrous dichloromethane (1.0 mL). The reaction system was cooled to 0°C, and N,N-diisopropylethylamine (20 mg, 0.15 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (27 mg, 0.07 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 26 (22 mg) in a yield of 67%.

[0397] MS-ESI calculated value [M+H] + 538, the actual measured value is 538.

[0398] 1 H NMR(400MHz,DMSO-d6)δ8.52(d,J=8.8Hz,1H),7.793(d,J=10.8Hz,1H),7.789(s ,1H),5.90(d,J=16.4Hz,1H),5.63-5.54(m,1H),5.52-5.54(m,2H),5.41-5.32( m,1H),4.15-4.04(m,1H),3.28-3.03(m,2H),2.39(s,3H),2.38-2.25(m,2H),2. 22-2.07(m,2H),1.93-1.84(m,2H),1.12(d,J=6.4Hz,3H),0.85(t,J=7.2Hz,3H).

[0399] Example 27

[0400] first step

[0401] To a single-necked flask (25 mL) were added 26a (45 mg, 0.09 mmol) and (R)-3-hydroxybutyric acid (19 mg, 0.18 mmol) in sequence, followed by anhydrous dichloromethane (1.0 mL). The reaction system was cooled to 0°C, and N,N-diisopropylethylamine (30 mg, 0.23 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (42 mg, 0.11 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 27 (45 mg) in a yield of 91%.

[0402] MS-ESI calculated value [M+H] + 538, the actual measured value is 538.

[0403] 1H NMR(400MHz, DMSO-d6)δ8.50(d,J=8.4Hz,1H),7.80-7.72(m,2H),5.90(d,J=16.4 Hz,1H),5.62-5.53(m,1H),5.52-5.43(m,2H),5.35(d,J=20.0Hz,1H),4.12-4.00 (m,1H),3.27-3.05(m,2H),2.44-2.39(m,1H),2.38(s,3H),2.28-2.21(m,1H),2. 20-2.08(m,2H),1.93-1.83(m,2H),1.10(d,J=6.0Hz,3H),0.85(t,J=7.2Hz,3H).

[0404] Example 28

[0405] first step

[0406] To a single-necked flask (25 mL) were added 26a (25 mg, 0.05 mmol) and 3-hydroxycyclobutanecarboxylic acid (12 mg, 0.10 mmol) in sequence, followed by anhydrous dichloromethane (1.0 mL). The reaction system was cooled to 0°C, and N,N-diisopropylethylamine (17 mg, 0.13 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (23 mg, 0.06 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 28 (26 mg) in a yield of 92%.

[0407] MS-ESI calculated value [M+H] + 550, the actual measured value is 550.

[0408] 1 H NMR (400MHz, DMSO-d6) δ7.81-7.75(m,2H),5.90(d,J=16.4Hz,1H),5.60-5.53(m,1H),5.48(d,J=16.8Hz,1H),5.41(d,J=20.0Hz,1H),5 .30(d,J=20.0Hz,1H),4.00-3.89(m,1H),3.30-3.05(m,2H),2.38(s,3H),2.25-2.00(m,4H),1.94-1.83(m,2H),0.85(t,J=7.2Hz,3H).

[0409] Example 29

[0410] first step

[0411] To a single-necked flask (25 mL) were added 26a (30 mg, 0.06 mmol) and cis-3-hydroxycyclobutanecarboxylic acid (14 mg, 0.12 mmol) in sequence, followed by anhydrous dichloromethane (1.0 mL). The reaction system was cooled to 0°C, and N,N-diisopropylethylamine (20 mg, 0.15 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (27 mg, 0.07 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 29 (33 mg) in a yield of 99%.

[0412] MS-ESI calculated value [M+H] + 550, the actual measured value is 550.

[0413] 1 H NMR (400MHz, DMSO-d6) δ8.49(d,J=8.8Hz,1H),7.78(s,1H),7.77(d,J=10.4Hz ,1H),5.90(d,J=16.4Hz,1H),5.61-5.52(m,1H),5.48(d,J=16.8Hz,1H),5.40 (d,J=20.0Hz,1H),5.28(d,J=20.0Hz,1H),4.01-3.89(m,1H),3.30-3.05(m,2 H),2.38(s,3H),2.25-2.00(m,4H),1.94-1.82(m,2H),0.85(t,J=7.2Hz,3H).

[0414] Example 30

[0415] first step

[0416] To a single-necked flask (25 mL) were added 26a (30 mg, 0.06 mmol) and trans-3-hydroxycyclobutanecarboxylic acid (14 mg, 0.12 mmol) in sequence, followed by anhydrous dichloromethane (1.0 mL). The reaction system was cooled to 0°C, and N,N-diisopropylethylamine (20 mg, 0.15 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (27 mg, 0.07 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 30 (22 mg) in a yield of 65%.

[0417] MS-ESI calculated value [M+H] + 550, the actual measured value is 550.

[0418] 1 H NMR(400MHz,DMSO-d6)δ8.46(d,J=8.8Hz,1H),7.79(d,J=10.4Hz,1H),7.78 (s,1H),5.90(d,J=16.4Hz,1H),5.62-5.53(m,1H),5.47(d,J=16.4Hz,1H), 5.44-5.27(m,2H),4.42-4.32(m,1H),3.28-3.06(m,2H),3.00-2.90(m,1H) ,2.38(s,3H),2.25-1.98(m,4H),1.94-1.82(m,2H),0.85(t,J=7.2Hz,3H).

[0419] Example 31

[0420] first step

[0421] To a single-necked flask (25 mL) were added 26a (30 mg, 0.06 mmol) and 3-hydroxycyclobutanecarboxylic acid (17 mg, 0.12 mmol) in sequence, followed by anhydrous dichloromethane (1.0 mL). The reaction system was cooled to 0°C, and N,N-diisopropylethylamine (20 mg, 0.15 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (27 mg, 0.07 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 31 (32 mg) in a yield of 90%.

[0422] MS-ESI calculated value [M+H]+ 578, the actual measured value is 578.

[0423] 1 H NMR (400MHz, DMSO-d6) δ7.78(s,1H),7.75(d,J=11.2Hz,1H),5.89(d,J=16.8H z,1H),5.60-5.51(m,1H),5.50-5.37(m,2H),5.34-5.20(m,1H),3.40-3.32(m ,1H),3.28-3.05(m,2H),2.37(s,3H),2.21-2.11(m,2H),2.10-2.00(m,1H),1 .94-1.62(m,6H),1.60-1.35(m,2H),1.19-1.04(m,2H),0.85(t,J=7.2Hz,3H).

[0424] Example 32

[0425] first step

[0426] To a single-necked flask (25 mL) were added 26a (30 mg, 0.06 mmol) and trans-4-hydroxycyclohexanecarboxylic acid (17 mg, 0.12 mmol) in sequence, followed by anhydrous dichloromethane (1.0 mL). The reaction system was cooled to 0°C, and N,N-diisopropylethylamine (20 mg, 0.15 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (27 mg, 0.07 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 32 (25 mg) in a yield of 70%.

[0427] MS-ESI calculated value [M+H] + 578, the actual measured value is 578.

[0428] 1H NMR(400MHz,DMSO-d6)δ8.46(d,J=8.4Hz,1H),7.80-7.73(m,2H),5.90(d,J=16.8H z,1H),5.59-5.51(m,1H),5.47(d,J=16.4Hz,1H),5.43(d,J=19.6Hz,1H),5.27(d, J=19.6Hz,1H),3.27-3.05(m,2H),2.38(s,3H),2.22-2.11(m,2H),2.10-2.00(m,2 H),1.94-1.77(m,6H),1.58-1.42(m,2H),1.18-1.05(m,2H),0.85(t,J=7.2Hz,3H).

[0429] Example 33

[0430] first step

[0431] To a single-necked flask (25 mL) were added 26a (30 mg, 0.06 mmol) and cis-4-hydroxycyclohexanecarboxylic acid (17 mg, 0.12 mmol) in sequence, followed by anhydrous dichloromethane (1.0 mL). The reaction system was cooled to 0°C, and N,N-diisopropylethylamine (20 mg, 0.15 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (27 mg, 0.07 mmol) were added in sequence. The reaction system was stirred at 0°C for half an hour. The reaction solution was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain compound 33 (25 mg) in a yield of 79%.

[0432] MS-ESI calculated value [M+H] + 578, the actual measured value is 578.

[0433] 1H NMR (400MHz, DMSO-d6) δ8.41(d,J=8.8Hz,1H),7.78(s,1H),7.77(d,J=10.8Hz,1H) ,5.90(d,J=16.4Hz,1H),5.61-5.52(m,1H),5.47(d,J=16.4Hz,1H),5.44(d,J=20. 0Hz,1H),5.32(d,J=20.0Hz,1H),3.28-3.05(m,2H),2.38(s,3H),2.30-2.03(m,4H ),1.95-1.80(m,4H),1.73-1.63(m,2H),1.56-1.34(m,4H),0.85(t,J=7.2Hz,3H).

[0434] Biological activity test

[0435] Cell proliferation inhibition assay

[0436] KPL-4 tumor cells in the logarithmic growth phase were taken and resuspended in fresh RPMI1640 culture medium. The cells were counted and the cell suspension was adjusted to 2×10 4 Cells / mL. The cell suspension was inoculated into a 96-well cell culture plate at 100 μL / well and cultured overnight in a carbon dioxide incubator (37°C, 5% CO2). The next day, one of the 96-well plates seeded with cells was removed and equilibrated to room temperature. 100 μL of CellTiter-Glo reagent (Promega, USA), which had been pre-equilibrated to room temperature and mixed thoroughly, was added to each well of the test plate. After incubation in the dark for 30 minutes, the luminescence value was read on a microplate reader (recorded as the G0 value). Another parallel plate was taken and different concentrations of the test compound or DMSO (final concentration 0.5%) were added to the corresponding wells of the test plate. After incubation in a carbon dioxide incubator for 72 hours, the test plate was equilibrated to room temperature and the cell activity was detected using the CellTiter-Glo reagent, which was recorded as the G3 value.

[0437] DXd was used as the positive control compound, and its structure is as follows:

[0438] The cell proliferation rate was calculated according to the following formula: Cell proliferation rate (%) = (average value of G3 in the test compound wells - average value of G0) / (average value of G3 in the DMSO control wells - average value of G0) * 100. Graphpad Prism software was used to fit the inhibition curve and calculate the GI 50 Values ​​(see table below).

[0439] Table 1: Cell proliferation inhibition assay results Note: a It represents the average of 2 measurements.

[0440] Table 2: Cell proliferation inhibition assay results

[0441] The results showed that several compounds of the present invention exhibited high anti-proliferation activity in the aforementioned tumor cell proliferation inhibition assay, significantly superior to that of DXd. Due to their excellent anti-proliferation activity, the compounds of the present invention can be used as tumor therapeutics or as toxin molecules for the preparation of antibody-drug conjugates for tumor treatment.

[0442] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. A compound represented by the following formula (I), or a pharmaceutically acceptable salt or hydrate thereof: in, n is 0 or 1; X is selected from the following group: N or CR 0 ; R 0 Selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, a C1-C8 alkyl group, a C1-C8 alkoxy group, OH, NH2, N3 or NO2; R 1 Selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, a C1-C8 alkyl group, a C1-C8 alkoxy group, a C1-C8 haloalkyl group, a C1-C8 haloalkoxy group, N3, NO2, NH2, NH-OH, -NR'R", -COOR', -CONR'R", -NHR"'NR'R", wherein R', R" and R'" are each independently selected from the group consisting of hydrogen, an alkyl group, an aryl group, an arylalkyl group, an acyl group, an alkoxycarbonyl group, and an aryloxycarbonyl group; R 2 、R 3 、R 4 、R 5 and R 6 Each is independently selected from the following groups: hydrogen atom, deuterium atom, halogen, hydroxyl, cyano, NH2, NO2, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C1-C8 alkoxy, substituted or unsubstituted C1-C8 alkylthio, substituted or unsubstituted C1-C8 deuterated alkyl, -(CH2) m tri(C1-C4alkyl)silyl, -(CH2) m (C3-C8 cycloalkyl), -(CH2) m (3-12 membered heterocyclic group), -(CH2) m N(R 7 )2、-(CH2) m S(CH2) p R 7 、-(CH2) m S(O)(CH2) p R 7 、-(CH2) m S(O)2(CH2) p R 7 、-(CH2) m NH(CH2) p R 7 、-(CH2) m NHC(O)(CH2) p R 7 、-(CH2) m OC(O)(CH2) p R 7 、-(CH2) m C(O)(CH2) p R 7 , -CH=N(OtBu); wherein m and p are each independently 0, 1, 2, 3 or 4; Or, R 2 and R 3 Together with the carbon atom to which it is attached, it forms a substituted or unsubstituted C5-C8 carbocyclic ring or a substituted or unsubstituted 5-12 membered heterocyclic ring; Or, R 2 and R 3 The carbon atoms connected thereto together form a structure selected from the group consisting of: unsubstituted or substituted with one or more R a substituted saturated or unsaturated 5-6 membered carbon ring, unsubstituted or replaced by one or more R a a substituted saturated or unsaturated 5-6 membered heterocycle; Or, R 3 and R 4 , or R 4 and R 5 The carbon atoms connected thereto together form a structure selected from the group consisting of: unsubstituted or substituted with one or more R a substituted saturated or unsaturated 5-12 membered carbon ring, unsubstituted or replaced by one or more R a substituted saturated or unsaturated 5-12 membered heterocycle; said R a is a substituted or unsubstituted substituent selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, a nitrile group, a nitro group, a hydroxyl group, an amino group, a C1-C6 alkyl group-NH-, a (C1-C6 alkyl)2N-, a C1-C6 alkyl group, a C2-C6 alkenyl group, a C2-C6 alkynyl group, a C1-C6 alkoxy group, an allyl group, a benzyl group, a C6-C 12 Aryl, C1-C6 alkoxy-C1-C6 alkyl, C1-C6 alkoxy-carbonyl, phenoxycarbonyl, C2-C6 alkynyl-carbonyl, C2-C6 alkenyl-carbonyl, C3-C6 cycloalkyl-carbonyl, C1-C6 alkyl-sulfonyl, phenyl, 5-7 membered heteroaryl, C3-C8 cycloalkyl, 3-12 membered heterocyclyl, -(CH2) m N(R 7 )2、-(CH2) m S(CH2) p R 7 、-(CH2) m S(O)(CH2) p R 7 、-(CH2) m S(O)2(CH2) p R 7 、-(CH2) m NH(CH2) p R 7 、-(CH2) m NHC(O)(CH2) p R 7 、-(CH2) m OC(O)(CH2) p R 7 、-(CH2) m C(O)(CH2) p R 7 ; wherein m and p are each independently 0, 1, 2, 3 or 4, preferably 0, 1 or 2; Each R 7 Each is independently selected from the following group: a hydrogen atom, a deuterium atom, a halogen, a substituted or unsubstituted C1-C8 alkyl group, a C1-C8 haloalkyl group, a C1-C8 deuterated alkyl group, a substituted or unsubstituted C1-C8 alkoxy group, a hydroxyl group, an amino group, a cyano group, a nitro group, a mercapto group, a substituted or unsubstituted C1-C8 alkylene group-OH, a substituted or unsubstituted C1-C8 alkylene group-NH2, SO2Me, -OC(O)(substituted or unsubstituted C1-C4 alkyl group), -C(O)(substituted or unsubstituted C1-C4 alkyl group), a substituted or unsubstituted phenyl group, a substituted or unsubstituted 5-7 membered heteroaryl group, a substituted or unsubstituted C3-C8 cycloalkyl group, a substituted or unsubstituted 3-12 membered heterocyclyl group; Unless otherwise specified, each of the above groups may be substituted by a substituent selected from the following groups: deuterium atom, halogen, nitrile, nitro, hydroxyl, amino, C1-C6 alkyl-NH-, (C1-C6 alkyl)2N-, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C2-C6 alkenyl, halogenated C2-C6 alkynyl, halogenated C1-C6 alkoxy, allyl, benzyl, C6-C 12 Aryl, C1-C6 alkoxy-C1-C6 alkyl, C1-C6 alkoxy-carbonyl, phenoxycarbonyl, C2-C6 alkynyl-carbonyl, C2-C6 alkenyl-carbonyl, C3-C6 cycloalkyl-carbonyl, C1-C6 alkyl-sulfonyl, phenyl, 5-7 membered heteroaryl, C3-C8 cycloalkyl, 3-12 membered heterocyclyl; And the compound is not a structure selected from the following group:

2. The compound according to claim 1, or a pharmaceutically acceptable salt or hydrate thereof, wherein The compound of formula I has a structure shown in formula II or formula III below:

3. The compound according to any one of claims 1 to 2, or a pharmaceutically acceptable salt or hydrate thereof, characterized in that: The compound of formula I has a structure shown in the following formula IV or formula V:

4. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt or hydrate thereof, characterized in that: The compound of formula I has a structure shown in the following formula VI or formula VII:

5. The compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt or hydrate thereof, characterized in that: The R 4 Selected from the following groups: hydrogen atom, deuterium atom, halogen, hydroxyl, cyano, NH2, NO2, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C1-C8 alkoxy, substituted or unsubstituted C1-C8 alkylthio, substituted or unsubstituted C1-C8 deuterated alkyl, -(CH2) m (C3-C8 cycloalkyl), -(CH2) m (3-12 membered heterocyclic group), -(CH2) m N(R 7 )2、-(CH2) m S(O)(CH2) p R 7 、-(CH2) m S(O)2(CH2) p R 7 、-(CH2) m NH(CH2) p R 7 ; wherein m and p are each independently 0, 1 or 2, R 7 is defined as above; R 5 Selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, NH2, OH, a substituted or unsubstituted C1-C8 alkyl group, a substituted or unsubstituted C1-C8 alkoxy group; Or, R 4 and R 5 The carbon atoms connected thereto together form a structure selected from the group consisting of: unsubstituted or substituted with one or more R a substituted saturated or unsaturated 5-6 membered carbon ring, unsubstituted or replaced by one or more R a Substituted saturated or unsaturated 5-6 membered heterocyclic ring; wherein R a The definition of is as described above.

6. The compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt or hydrate thereof, characterized in that: R 2 and R 3 are each independently selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, NH2, a substituted or unsubstituted C1-C8 alkyl group, a substituted or unsubstituted C1-C8 deuterated alkyl group, -(CH2) m (C3-C6 cycloalkyl), -(CH2) m (3-6 membered heterocyclic group), -(CH2) m N(R 7 )2、-(CH2) m OC(O)R 7 ; wherein m is 0, 1, 2, 3 or 4, R 7 is defined as above; Or, R 2 and R 3 The carbon atoms connected thereto together form a structure selected from the group consisting of: unsubstituted or substituted with one or more R a substituted saturated or unsaturated 5-6 membered carbon ring, unsubstituted or replaced by one or more R a Substituted saturated or unsaturated 5-6 1-membered heterocyclic ring; wherein R a The definition of is as described above.

7. The compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt or hydrate thereof, characterized in that: R 1 and R 6 are each independently a hydrogen atom.

8. The compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt or hydrate thereof, characterized in that: The R 4 and R 5 Each is independently selected from the following group: a hydrogen atom, a deuterium atom, a halogen, a hydroxyl group, NH2, a substituted or unsubstituted C1-C4 alkyl group, a substituted or unsubstituted C1-C4 alkoxy group, and a cyclopropyl group; Or, R 4 and R 5 The carbon atoms to which it is attached together form an unsubstituted or substituted carbon atom or carbon atoms. a Substituted oxa 5-6 membered heterocycle; wherein R a The definition of is as described above.

9. The compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt or hydrate thereof, characterized in that: R 2 Selected from the group consisting of: deuterium atom, halogen, NH2, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C1-C8 deuterated alkyl, -(CH2) m (C3-C6 cycloalkyl), -(CH2) m (3-6 membered heterocyclic group), -(CH2) m N(R 7 )2、-(CH2) m OC(O)R 7 ; wherein m is 0, 1, 2, 3 or 4; R 3 Selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, a substituted or unsubstituted C1-C8 alkyl group, a substituted or unsubstituted C1-C8 deuterated alkyl group, -(CH2) m (C3-C6 cycloalkyl), -(CH2) m (3-6 membered heterocyclic group), -(CH2) m N(R 7 )2、-(CH2) m OC(O)R 7 ; wherein m is 0, 1, 2, 3 or 4; Or, R 2 and R 3 The carbon atoms connected thereto together form a structure selected from the group consisting of: unsubstituted or substituted with one or more R a substituted saturated or unsaturated 5-6 membered carbon ring, unsubstituted or replaced by one or more R a a substituted saturated or unsaturated 5-6 membered heterocycle; R 4 Selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, a hydroxyl group, a cyano group, NH2, NO2, a substituted or unsubstituted C1-C8 alkyl group, a substituted or unsubstituted C1-C8 alkoxy group; R 5 Selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, and a substituted or unsubstituted C1-C8 alkyl group; Or, R 4 and R 5 connected to form unsubstituted or one or more R a A substituted structure selected from the group consisting of: -OCH2O- or -O(CH2)2O-; R 7 Selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, a substituted or unsubstituted C1-C8 alkyl group, a hydroxyl group, an amino group, a cyano group, a nitro group, and a mercapto group; Among them, R a The definition of is as described above.

10. The compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt or hydrate thereof, characterized in that: The compound has a structure shown in the following formula:

11. A pharmaceutical composition comprising a compound of formula I according to any one of claims 1 to 10, or a pharmaceutically acceptable salt or hydrate thereof, and one or more pharmaceutically acceptable excipients, diluents or carriers.

12. The use of a compound of formula I according to any one of claims 1 to 10, characterized in that Used for preparing a pharmaceutical composition for treating diseases related to tumor cell proliferation.

13. The use of a compound of formula I according to any one of claims 1 to 10, characterized in that Used as a toxin in antibody-drug conjugates to prepare antibody-drug conjugates.

14. A compound represented by the following formula II-b or III-b: in, The definitions of the various groups are as described in claim 1.

15. The compound according to claim 14, wherein The compound has the structure described in Formula IV-b or Vb below:

16. A method for preparing a compound of formula I according to any one of claims 1 to 10, characterized in that: The method comprises the steps of: In an inert solvent, the compound of formula Ia is reacted with the compound of formula Ib to obtain the compound of formula I.

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