Fused ring compounds, pharmaceutical composition comprising same and use thereof

Abstract

Disclosed are compounds of formula (I) useful as Werner helicase (WRN) inhibitors, a pharmaceutical composition comprising same, and the use thereof in the prevention or treatment of cancers characterized by microsatellite instability-high (MSI-H) or defective mismatch repair (dMMR).

Description

Condensed ring compound, pharmaceutical composition containing the same and use thereof Field of the Invention

[0001] The present invention relates to a fused ring compound, a pharmaceutical composition containing the same, and use thereof for preventing or treating diseases.

[0002] Background of the Invention

[0003] Microsatellite instability (MSI) is a genomic damage caused by mismatch repair deficiency (dMMR). Although progress has been made in treating microsatellite high instability (MSI-H) cancers, tumor evolution and drug resistance remain the main causes of treatment failure and death in cancer patients. For example, in patients with dMMR colorectal cancer treated with PD-1 and PD-L1 checkpoint inhibitors, about half of the patients experienced primary drug resistance. See, for example, Overman MJ et al., J Clin Oncol. 2018 Mar 10; 36(8): 773-779. For patients who are difficult to treat with currently available treatments, there is still a clinical need for new treatment options that have not yet been met.

[0004] Werner helicase (WRN) is considered to be a synthetic lethal target for MSI-H cancer (see, for example, Chan EM et al., Nature.2019Apr; 568(7753): 551-556). WRN is one of the members of the DNA helicase RecQ family and plays an important role in maintaining genomic stability, DNA repair, replication, transcription and telomere maintenance. In the study of the WRN dependency mechanism, it was found that dinucleotide TA repeats were amplified on a large scale in MSI cells. These amplified TA repeats formed secondary DNA structures, requiring WRN helicase to unwind (see, for example, van Wietmarschen N et al., Nature.2020 Oct; 586(7828): 292-298). In the absence of WRN (or when WRN helicase is inhibited), the TA repeats amplified in MSI cells are cut by nucleases and eventually lead to chromosome breakage. Therefore, inhibition of WRN helicase is an effective strategy for treating cancers characterized by microsatellite instability-high (MSI-H) or mismatch repair deficiency (dMMR), including colorectal, gastric, or endometrial cancer.

[0005] SUMMARY OF THE INVENTION

[0006] The present application provides compounds used as WRN inhibitors, which can be used to prevent or treat cancers characterized by microsatellite high instability (MSI-H) or mismatch repair deficiency (dMMR). In addition, the compounds of the present invention also have excellent properties such as good physicochemical properties (such as solubility, physical and / or chemical stability), good pharmacokinetic properties (such as improved bioavailability, good metabolic stability, suitable half-life and duration of action), good safety (lower toxicity (such as reduced cardiotoxicity) and / or fewer side effects), and are less likely to develop drug resistance.

[0007] One aspect of the present invention provides a compound or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein the compound has the structure of Formula (I):

[0008] in:

[0009] Selected from where R 1 Connected to the position marked with "*" in the above group, R 2 Connected to the position marked with "**" in the above group, R 3 Connected to the position marked with "***" in the above group, and R 4 Connected to the position marked with "****" in the above group;

[0010] R 1 Selected from-NR 11 R 12 、

[0011] R 2 for

[0012] R 4 for

[0013] R 3 、R 11 、R 12 、R 13 、R 21 、R 22 、R 23 and R 24 Each occurrence is independently selected from H, halogen, -OH, -NH2, -CN, -NO2, -SF5, C 1-6 Alkyl, deuterated C 1-6 Alkyl, halogenated C 1-6Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl, C 6-12 Aralkyl, -C(=O)R a 、-OC(=O)R a 、-C(=O)OR a 、-OR a 、-SR a 、-S(=O)R a 、-S(=O)2R a 、-S(=O)2NR a R b 、-NR a R b 、-C(=O)NR a R b 、-NR a -C(=O)R b 、-NR a -C(=O)OR b 、-NR a -S(=O)2-R b 、-NR a -C(=O)-NR a R b 、-P(=O)R a R b 、-C 1-6 Alkylene-R a 、-C 1-6 Alkylene-OR a 、-C 1-6 Alkylene-NR a R b 、-OC 1-6 Alkylene-NR a R b 、(-C 3-6 Cycloalkylene)-CN and (-C 3-6 Cycloalkylene)-C 1-6 alkyl halide;

[0014] R 14 Each occurrence is independently selected from -L 1 -(C 3-6 Cycloalkyl), -L 1 -(3-10 membered heterocyclic group), -L 1 -(C 6-10 aryl) and -L 1 -(5-14 membered heteroaryl);

[0015] R41 Selected from C 3-6 hydrocarbon ring, 3-10 membered heterocyclic ring, C 6-10 aromatic rings and 5-14 membered heteroaromatic rings;

[0016] Ring C, Ring D, Ring E, Ring X and Ring Z are each independently selected from C 3-6 hydrocarbon ring, 3-10 membered heterocyclic ring, C 6-10 Aromatic rings and 5-14 membered heteroaromatic rings;

[0017] Ring Y is absent or selected from C 3-6 Hydrocarbon ring, 3-10 membered heterocycle, C 6-10 aromatic rings and 5-14 membered heteroaromatic rings; when ring Y is absent, R 24 It does not exist either;

[0018] L 1 Selected from direct bond, -O-, -C(=O)-, -NRC(=O)-, -S-, -S(=O)-, -S(=O)2-, C 1-6 Alkylene and -O-(C 1-6 alkylene)-;

[0019] L 2 Selected from -O-, -C(=O)-, -NRC(=O)-, -S-, -S(=O)-, -S(=O)2-, C 1-6 Alkylene and -O-(C 1-6 alkylene)-;

[0020] R, R a and R b Each occurrence is independently selected from H, C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl and C 6-12 Aralkyl;

[0021] The above alkylene, alkyl, alkenyl, alkynyl, cycloalkylene, cycloalkyl, hydrocarbon ring, heterocyclyl, heterocycle, aryl, aromatic ring, heteroaryl, heteroaromatic ring and aralkyl groups are each optionally substituted with one or more substituents independently selected from the group consisting of halogen, -OH, =O, =CF2, -NH2, -CN, -NO2, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl, C 6-12 Aralkyl, -C(=O)R c 、-OC(=O)R c 、-C(=O)OR c、-OR c 、-SR c 、-S(=O)R c 、-S(=O)2R c 、-S(=O)2NR c R d 、-NR c R d 、-C(=O)NR c R d 、-NR c -C(=O)R d 、-NR c -C(=O)OR d 、-NR c -S(=O)2-R d 、-NR c -C(=O)-NR c R d 、-C 1-6 Alkylene-OR c 、-C 1-6 Alkylene-NR c R d and -OC 1-6 Alkylene-NR c R d The alkylene, alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and aralkyl groups are further optionally substituted by one or more substituents independently selected from the following: halogen, -OH, =O, -C(=O)O-tert-butyl, -NH2, -CN, -NO2, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl, C 6-12 Aralkyl, -OC 1-6 Alkyl and -C 1-6 Alkylene-OC 1-6 alkyl;

[0022] R c and R d Each occurrence is independently selected from H, C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl and C 6-12 Aralkyl, said alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and aralkyl further optionally substituted by one or more substituents independently selected from the following: halogen, -OH, =O, -C(=O)O-tert-butyl, -NH2, -CN, -NO2, C 1-6 Alkyl, C1-6 Halogenated alkyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl, C 6- 12 Aralkyl and -C 1-6 Alkylene-OC 1-6 alkyl;

[0023] n, p and q are each independently an integer selected from 1, 2 or 3; and

[0024] m is an integer of 0 or 1.

[0025] Another aspect of the present invention provides a pharmaceutical composition comprising a prophylactically or therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof and one or more pharmaceutically acceptable carriers.

[0026] Another aspect of the present invention provides use of a compound of the present invention or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, or a pharmaceutical composition of the present invention in the preparation of a medicament for use as a WRN inhibitor.

[0027] Another aspect of the present invention provides a compound of the present invention, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, or a pharmaceutical composition of the present invention, for use as a WRN inhibitor.

[0028] Another aspect of the present invention provides a method for preventing or treating cancer (preferably a cancer characterized by microsatellite high instability (MSI-H) or mismatch repair deficiency (dMMR)), which comprises administering to a subject in need thereof an effective amount of a compound of the present invention or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, or a pharmaceutical composition of the present invention.

[0029] Detailed Description of the Invention

[0030] definition

[0031] Unless otherwise defined below, all technical and scientific terms used herein are intended to have the same meaning as those commonly understood by those skilled in the art. References to technology used herein are intended to refer to technology commonly understood in the art, including variations of technology or substitutions of equivalent technology that would be apparent to those skilled in the art. While it is believed that the following terms are well understood by those skilled in the art, the following definitions are set forth to better explain the present invention.

[0032] "Zwitterion" or "zwitterionic form" refers to a compound that contains both positively and negatively charged functional groups. In some instances, certain groups are inherently non-zwitterionic, but in a tautomer in which a proton is shifted, the group is in zwitterionic form. For example, when R 41 When the group is a pyridine ring substituted with at least one -OH group, it may be in the following zwitterionic form (c) or non-zwitterionic form (d) or a mixture of the two forms:

[0033] where R 100 、R 101 、R 102 represents a substituent as defined herein;

[0034] For example, when R 41 When the group is a pyrimidine ring substituted with at least one -OH group, it may be in the following zwitterionic form (a) or (b) or non-zwitterionic form (e), or a mixture of any two or three thereof:

[0035] where R 100 、R 102 represents a substituent as defined herein.

[0036] The terms "comprises," "comprising," "having," "containing," or "involving," and other variations thereof herein, are inclusive or open-ended and do not exclude additional unrecited elements or method steps.

[0037] As used herein, the term "alkylene" refers to a saturated divalent hydrocarbon group, preferably a saturated divalent hydrocarbon group having 1, 2, 3, 4, 5 or 6 carbon atoms, such as methylene, ethylene, propylene or butylene.

[0038] As used herein, the term "alkyl" is defined as a straight or branched chain saturated aliphatic hydrocarbon. In some embodiments, the alkyl group has 1 to 12, such as 1 to 6, carbon atoms. For example, as used herein, the term "C 1-6 "Alkyl" refers to a linear or branched group of 1 to 6 carbon atoms (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl or n-hexyl), which is optionally substituted with one or more (e.g., 1 to 3) suitable substituents such as halogen (in which case the group is referred to as "haloalkyl") (e.g., CF3, C2F5, CHF2, CH2F, CH2CF3, CH2Cl or -CH2CH2CF3, etc.). The term "C 1-4"Alkyl" refers to a linear or branched aliphatic hydrocarbon chain of 1 to 4 carbon atoms (ie, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl).

[0039] As used herein, the term "alkenyl" means a linear or branched monovalent hydrocarbon radical containing one or more double bonds and having 2 to 6 carbon atoms ("C 2-6 The alkenyl group is, for example, -CH=CH2, -CH2CH=CH2, -C(CH3)=CH2, -CH2-CH=CH-CH3, 2-pentenyl, 3-pentenyl, 4-pentenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 2-methyl-2-propenyl and 4-methyl-3-pentenyl. When the compound of the present invention contains an alkenyl group, the compound may be in the pure E (entgegen) form, the pure Z (zusammen) form or any mixture thereof. The term "alkenylene" is a corresponding divalent group, including, for example, "C 2-6 Alkenylene", "C 2-4 Specific examples include, but are not limited to, -CH=CH-, -CH2CH=CH-, -C(CH3)=CH-, butenylene, pentenylene, hexenylene, etc.

[0040] As used herein, the term "alkynyl" refers to a monovalent hydrocarbon radical containing one or more triple bonds, preferably having 2, 3, 4, 5 or 6 carbon atoms, such as ethynyl, 2-propynyl, 2-butynyl, 1,3-butadiynyl, etc. The alkynyl group is optionally substituted with one or more (such as 1 to 3) identical or different substituents. The term "alkynylene" is a corresponding divalent radical, including, for example, "C 2-8 Alkynylidene", "C 2-6 Alkynylidene", "C 2-4 Examples include, but are not limited to, The alkynylene group is optionally substituted with one or more (such as 1 to 3) identical or different substituents.

[0041] As used herein, the term "paracyclic" or "fused ring" refers to a ring system formed by two or more cyclic structures that share two adjacent atoms.

[0042] As used herein, the term "spirocycle" refers to a ring system formed by two or more cyclic structures that share one ring atom with each other.

[0043] As used herein, the term "bridged ring" refers to a ring system formed by two or more cyclic structures sharing two atoms that are not directly connected to each other.

[0044] As used herein, the terms "cycloalkylene", "cycloalkyl" and "hydrocarbon ring" refer to saturated (i.e., "cycloalkylene" and "cycloalkyl") or partially unsaturated (i.e., having one or more double bonds and / or triple bonds within the ring) monocyclic or polycyclic hydrocarbon rings (including spiro, fused or bridged ring systems) having, for example, 3-10 (suitably 3-8, more suitably 3-6) ring carbon atoms, including but not limited to (cyclo)propyl, (cyclo)butyl, (cyclo)pentyl, (cyclo)hexyl, (cyclo)heptyl, (cyclo)octyl, (cyclo)nonyl, (cyclo)hexenyl and the like.

[0045] As used herein, the term "cycloalkyl" refers to a saturated monocyclic or polycyclic (such as bicyclic) hydrocarbon ring (e.g., a monocyclic ring such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or a bicyclic ring, including spirocyclic, fused or bridged systems (such as bicyclo[1.1.1]pentyl, bicyclo[2.2.1]heptyl, bicyclo[3.2.1]octyl or bicyclo[5.2.0]nonyl, decalinyl, etc.), which is optionally substituted with one or more (such as one to three) suitable substituents. The cycloalkyl group has 3 to 15 carbon atoms. For example, the term "C 3-6 "Cycloalkyl" refers to a saturated monocyclic or polycyclic (such as bicyclic) hydrocarbon ring of 3 to 6 ring carbon atoms (for example cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl), which is optionally substituted by 1 or more (such as 1 to 3) suitable substituents, for example methyl substituted cyclopropyl.

[0046] As used herein, the term "heterocyclyl" (or "heterocycle") refers to a saturated or partially unsaturated monocyclic or bicyclic group having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms and one or more (e.g., one, two, three or four) heteroatoms selected from O, S, N and P in the ring, and the "heterocyclyl" (or "heterocycle") may contain -C(=O)- as a ring member. The heterocyclyl group may be attached to the rest of the molecule via the carbon atoms and / or heteroatoms, if present. In particular, a 3-10 membered heterocyclyl group is a group having 3-10 carbon atoms and heteroatoms in the ring, such as, but not limited to, an oxiranyl, an aziridinyl, an azetidinyl, an oxetanyl, a tetrahydrofuranyl, a dioxolinyl, a pyrrolidinyl, a pyrrolidonyl, an imidazolidinyl, a pyrazolidinyl, a pyrrolinyl, a tetrahydropyranyl, a piperidinyl, a morpholinyl, a dithianyl, a thiomorpholinyl, a piperazinyl or a trithianyl group.

[0047] As used herein, the term "heterocyclyl" (or "heterocycle") encompasses a parallel ring structure, and the connection point of the parallel ring structure to the other groups can be on any ring in the parallel ring structure. Therefore, the heterocyclyl of the present invention also includes but is not limited to heterocyclyl and heterocyclyl, heterocyclyl and cycloalkyl, monoheterocyclyl and monoheterocyclyl, monoheterocyclyl and monocycloalkyl, aryl and heterocyclyl, heteroaryl and heterocyclyl, such as 3-7 membered (mono) heterocyclyl and 3-7 membered (mono) heterocyclyl, 3-7 membered (mono) heterocyclyl and (mono) cycloalkyl, 3-7 membered (mono) heterocyclyl and C 4-6 (mono)cycloalkyl, C 6-10 Aryl and 3-7 membered heterocyclic group, 5-6 membered heteroaryl and 3-7 membered heterocyclic group, examples of which include but are not limited to pyrrolidinyl and cyclopropyl, cyclopentyl and aziridine, pyrrolidinyl and cyclobutyl, pyrrolidinyl and pyrrolidinyl, pyrrolidinyl and piperidinyl, pyrrolidinyl and piperazinyl, piperidinyl and morpholinyl,

[0048] As used herein, the term "heterocyclyl" (or "heterocycle") encompasses bridged heterocyclyls (bridged heterocycle) and spiro heterocyclyls (spiro heterocycle).

[0049] As used herein, the term "bridged heterocycle" refers to a cyclic structure containing one or more (e.g., 1, 2, 3, or 4) heteroatoms (e.g., oxygen atoms, nitrogen atoms, and / or sulfur atoms) formed by two rings sharing two ring atoms that are not directly connected, including but not limited to 7-10 membered bridged heterocycles, 8-10 membered bridged heterocycles, 7-10 membered nitrogen-containing bridged heterocycles, 7-10 membered oxygen-containing bridged heterocycles, 7-10 membered sulfur-containing bridged heterocycles, etc., for example The "nitrogen-containing bridged heterocycle", "oxygen-containing bridged heterocycle" and "sulfur-containing bridged heterocycle" optionally further contain one or more other heteroatoms selected from oxygen, nitrogen and sulfur.

[0050] As used herein, the term "spiroheterocycle" refers to a cyclic structure containing one or more (e.g., 1, 2, 3, or 4) heteroatoms (e.g., oxygen atoms, nitrogen atoms, sulfur atoms) formed by two or more rings sharing a ring atom, including but not limited to 5-10 membered spiroheterocycle, 6-10 membered spiroheterocycle, 6-10 membered nitrogen-containing spiroheterocycle, 6-10 membered oxygen-containing spiroheterocycle, 6-10 membered sulfur-containing spiroheterocycle, etc., for example The "nitrogen-containing spiroheterocycle", "oxygen-containing spiroheterocycle" and "sulfur-containing spiroheterocycle" optionally further contain one or more other heteroatoms selected from oxygen, nitrogen and sulfur. The term "6-10 membered nitrogen-containing spiroheterocyclyl" refers to a spiroheterocyclyl containing 6-10 ring atoms, at least one of which is a nitrogen atom.

[0051] As used herein, the terms "arylene" and "aromatic ring" refer to an all-carbon monocyclic or fused-ring polycyclic aromatic group having a conjugated π electron system. For example, as used herein, the term "C 6-10 (E)aryl" and "C 6-10 The term "aromatic ring" means an aromatic group containing 6 to 10 carbon atoms, such as (ene)phenyl (phenyl ring) or (ene)naphthyl (naphthalene ring). The (ene)aryl group and the aromatic ring are optionally substituted by one or more (such as one to three) suitable substituents (e.g., halogen, -OH, -CN, -NO2, C 1-6 alkyl, etc.) substituted.

[0052] The term "aralkyl" refers to an alkyl group substituted with an aryl group, wherein the aryl group and the alkyl group are as defined herein. Typically, the aryl group may have 6 to 14 carbon atoms, and the alkyl group may have 1 to 6 carbon atoms. Exemplary aralkyl groups include, but are not limited to, benzyl, phenylethyl, phenylpropyl, and phenylbutyl.

[0053] As used herein, the terms "heteroaryl(ene)" and "heteroaromatic ring" refer to a monocyclic, bicyclic or tricyclic aromatic ring system having 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms, in particular 1 or 2 or 3 or 4 or 5 or 6 or 9 or 10 carbon atoms, and which contains at least one heteroatom which may be identical or different (the heteroatom being for example oxygen, nitrogen or sulfur) and, in each case, may additionally be benzo-fused. In particular, “heteroaryl” or “heteroaryl ring” is selected from thienyl (ring), furanyl (ring), pyrrolyl (ring), oxazolyl (ring), thiazolyl (ring), imidazolyl (ring), pyrazolyl (ring), isoxazolyl (ring), isothiazolyl (ring), oxadiazolyl (ring), triazolyl (ring), thiadiazolyl (ring), etc., and benzo derivatives thereof; or pyridinyl (ring), pyridazinyl (ring), pyrimidinyl (ring), pyrazinyl (ring), triazinyl (ring), etc., and benzo derivatives thereof.

[0054] As used herein, the term "halo" or "halogen" group is defined to include F, Cl, Br, or I.

[0055] The term "alkylthio" as used herein, means an alkyl group, as defined above, appended to the parent molecular moiety through a sulfur atom. 1-6 Representative examples of alkylthio include, but are not limited to, methylthio, ethylthio, tert-butylthio, and hexylthio.

[0056] As used herein, the term "nitrogen-containing heterocycle" refers to a saturated or partially unsaturated monocyclic or bicyclic group having 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13 carbon atoms and at least one nitrogen atom in the ring, which may optionally further contain one or more (e.g., one, two, three or four) ring members selected from N, O, S, S=O and S(=O); the nitrogen-containing heterocycle is connected to the rest of the molecule via any ring member. The nitrogen-containing heterocycle is preferably a saturated nitrogen-containing monocyclic ring. In particular, the 3- to 14-membered nitrogen-containing heterocycle is a group having 3-14 carbon atoms and heteroatoms (at least one of which is a nitrogen atom) in the ring, including but not limited to a three-membered nitrogen-containing heterocycle (such as aziridine), a four-membered nitrogen-containing heterocycle (such as azetidinyl), a five-membered nitrogen-containing heterocycle (such as pyrrolyl, pyrrolidinyl (pyrrolidine ring), pyrrolinyl, pyrrolidonyl, imidazolyl, imidazolidinyl, imidazolinyl, pyrazolyl, pyrazolinyl), a six-membered nitrogen-containing heterocycle (such as piperidinyl (piperidine ring), morpholinyl, thiomorpholinyl, piperazinyl), a seven-membered nitrogen-containing heterocycle, etc.

[0057] The term "substituted" means that one or more (e.g., one, two, three, or four) hydrogen atoms on the designated atom are replaced with a group selected from the indicated group, provided that the designated atom's normal valence is not exceeded in the current context and that the substitution results in a stable compound. Combinations of substituents and / or variables are permissible only if such combinations result in stable compounds.

[0058] If a substituent is described as being "optionally substituted," the substituent may be (1) unsubstituted or (2) substituted. If a carbon of a substituent is described as being optionally substituted with one or more of the substituents listed, one or more hydrogens on the carbon (to the extent of any hydrogens present) may be replaced, individually and / or collectively, with independently selected optional substituents. If a nitrogen of a substituent is described as being optionally substituted with one or more of the substituents listed, one or more hydrogens on the nitrogen (to the extent of any hydrogens present) may each be replaced with an independently selected optional substituent.

[0059] If substituents are described as being "independently selected" from a group, each substituent is selected independently of the other. Thus, each substituent may be the same as or different from another (other) substituent.

[0060] As used herein, the term "one or more" means 1 or more than 1, such as 2, 3, 4, 5 or 10, where reasonable.

[0061] Unless otherwise indicated, as used herein, the point of attachment of a substituent may be from any suitable position of the substituent.

[0062] When a bond to a substituent is shown to pass through a bond connecting two atoms in a ring, then such substituent may be bonded to any ring atom in the substitutable ring.

[0063] The present invention also includes all pharmaceutically acceptable isotopically labeled compounds, which are identical to the compounds of the present invention except that one or more atoms are replaced by an atom having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number prevalent in nature. Examples of isotopes suitable for inclusion in the compounds of the present invention include, but are not limited to, isotopes of hydrogen (e.g., deuterium (D, 2 H), tritium (T, 3 H)); carbon isotopes (e.g. 11 C. 13 C and 14 C); isotopes of chlorine (e.g. 36 Cl); isotopes of fluorine (e.g. 18 F); isotopes of iodine (such as 123 I and 125 I); isotopes of nitrogen (e.g. 13 N and 15 N); oxygen isotopes (e.g. 15 O. 17 O and 18 O); isotopes of phosphorus (such as 32 P); and sulfur isotopes (e.g. 35 S). Certain isotopically labeled compounds of the invention (e.g., those incorporating radioactive isotopes) are useful in drug and / or substrate tissue distribution studies (e.g., assays). The radioactive isotope tritium (i.e., 3 H) and carbon-14 (i.e. 14 C) are particularly useful for this purpose because they are easy to incorporate and easy to detect. 11 C. 18 F. 15 O and 13 N) substitution can be used to examine substrate receptor occupancy in positron emission tomography (PET) studies. Isotopically labeled compounds of the present invention can be prepared by methods analogous to those described in the accompanying schemes and / or examples and preparations by using appropriate isotopically labeled reagents instead of the non-labeled reagents previously employed. Pharmaceutically acceptable solvates of the present invention include those in which the crystallization solvent is isotopically substituted, for example, D2O, acetone-d6 or DMSO-d6.

[0064] The term "stereoisomer" refers to an isomer formed due to at least one asymmetric center. In compounds with one or more (e.g., one, two, three, or four) asymmetric centers, racemic mixtures, single enantiomers, diastereomeric mixtures, and individual diastereomers can be produced. Specific individual molecules can also exist as geometric isomers (cis / trans). Similarly, the compounds of the present invention can exist as mixtures of two or more structurally different forms in rapid equilibrium (commonly referred to as tautomers). Representative examples of tautomers include keto-enol tautomers, phenol-ketone tautomers, nitroso-oxime tautomers, imine-enamine tautomers, etc. It is to be understood that the scope of this application encompasses all such isomers or mixtures thereof in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%).

[0065] In this article, solid lines can be used Solid wedge or virtual wedge The carbon-carbon bonds of the compounds of the present invention are depicted. The use of solid lines to depict bonds to asymmetric carbon atoms is intended to indicate that all possible stereoisomers at that carbon atom are included (e.g., specific enantiomers, racemic mixtures, etc.). The use of solid or dashed wedges to depict bonds to asymmetric carbon atoms is intended to indicate that the indicated stereoisomers exist. When present in a racemic mixture, solid and dashed wedges are used to define relative stereochemistry, not absolute stereochemistry. Unless otherwise indicated, the compounds of the present invention are intended to exist as stereoisomers, including cis and trans isomers, optical isomers (e.g., R and S enantiomers), diastereomers, geometric isomers, rotational isomers, conformational isomers, atropisomers, and mixtures thereof. The compounds of the present invention may exhibit more than one type of isomerism and consist of mixtures thereof (e.g., racemic mixtures and diastereomeric pairs).

[0066] Atropisomers are compounds that can be separated into rotationally restricted isomers.

[0067] It should also be understood that certain compounds of the present invention may be used therapeutically in free form or, where appropriate, in the form of pharmaceutically acceptable derivatives thereof. In the present invention, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, esters, solvates, metabolites, or prodrugs that, upon administration to a patient in need thereof, are capable of directly or indirectly providing a compound of the present invention or a metabolite or residue thereof. Therefore, when reference is made herein to a "compound of the present invention," such various derivative forms of the compound are also intended to be encompassed.

[0068] Pharmaceutically acceptable salts of the compounds of the present invention include acid addition salts and base addition salts thereof.

[0069] For a review of suitable salts see Stahl and Wermuth, “Handbook of Pharmaceutical Salts: Properties, Selection, and Use” (Wiley-VCH, 2002). Methods for preparing pharmaceutically acceptable salts of the compounds of the invention are known to those skilled in the art.

[0070] As used herein, the term "ester" refers to esters derived from the compounds of the general formulae herein, including physiologically hydrolyzable esters (which can be hydrolyzed under physiological conditions to release the compounds of the present invention in the form of free acid or alcohol). The compounds of the present invention themselves may also be esters.

[0071] The compounds of the present invention may exist in the form of solvates (preferably hydrates), wherein the compounds of the present invention contain a polar solvent as a structural element of the crystal lattice of the compound, in particular water, methanol or ethanol. The amount of polar solvent, in particular water, may be present in a stoichiometric or non-stoichiometric ratio.

[0072] Also included within the scope of the present invention are metabolites of the compounds of the present invention, i.e., substances formed in vivo upon administration of the compounds of the present invention. Such products may be produced, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, defatting, enzymatic hydrolysis, and the like of the administered compound. Thus, the present invention includes metabolites of the compounds of the present invention, including compounds produced by contacting a compound of the present invention with a mammal for a period of time sufficient to produce a metabolic product thereof.

[0073] The present invention further includes within its scope prodrugs of the compounds of the present invention, which are certain derivatives of the compounds of the present invention that may themselves have little or no pharmacological activity and can be converted into compounds of the present invention having the desired activity by, for example, hydrolytic cleavage when administered to the body or thereon. Typically, such prodrugs will be functional group derivatives of the compounds that are readily converted into the desired therapeutically active compounds in vivo. Additional information on the use of prodrugs can be found in "Pro-drugs as Novel Delivery Systems," Volume 14, ACS Symposium Series (T. Higuchi and V. Stella) and "Bioreversible Carriers in Drug Design," Pergamon Press, 1987 (E.B. Roche, ed., American Pharmaceutical Association). Prodrugs of the present invention can be prepared, for example, by replacing appropriate functional groups present in the compounds of the present invention with certain moieties known to those skilled in the art as "pro-moieties" (e.g., as described in "Design of Prodrugs," H. Bundgaard (Elsevier, 1985)).

[0074] The present invention also encompasses compounds of the present invention that contain protecting groups. During any process for preparing the compounds of the present invention, it may be necessary and / or desirable to protect sensitive or reactive groups on any of the molecules involved, thereby forming a chemically protected form of the compounds of the present invention. This can be achieved using conventional protecting groups, for example, those described in Protective Groups in Organic Chemistry, ed. JFW McOmie, Plenum Press, 1973; and TW Greene & P.GM Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991, which references are incorporated herein by reference. Protecting groups can be removed at an appropriate subsequent stage using methods known in the art.

[0075] As used herein, the term "about" means within ±10%, preferably within ±5%, and more preferably within ±2% of the stated numerical value.

[0076] Compound

[0077] In some embodiments, the present disclosure provides a compound or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound, or prodrug thereof, wherein the compound has the structure of Formula (I):

[0078] in:

[0079] Selected from where R 1 Connected to the position marked with "*" in the above group, R 2 Connected to the position marked with "**" in the above group, R 3 Connected to the position marked with "***" in the above group, and R 4 Connected to the position marked with "****" in the above group;

[0080] R 1 Selected from-NR 11 R 12 、

[0081] R 2 for

[0082] R 4 for

[0083] R 3 、R 11 、R 12 、R 13 、R 21 、R 22 、R 23 and R 24 Each occurrence is independently selected from H, halogen, -OH, -NH2, -CN, -NO2, -SF5, C 1-6 Alkyl, deuterated C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl, C 6-12 Aralkyl, -C(=O)R a 、-OC(=O)R a 、-C(=O)OR a 、-OR a 、-SR a 、-S(=O)R a、-S(=O)2R a 、-S(=O)2NR a R b 、-NR a R b 、-C(=O)NR a R b 、-NR a -C(=O)R b 、-NR a -C(=O)OR b 、-NR a -S(=O)2-R b 、-NR a -C(=O)-NR a R b 、-P(=O)R a R b 、-C 1-6 Alkylene-R a 、-C 1-6 Alkylene-OR a 、-C 1-6 Alkylene-NR a R b 、-OC 1-6 Alkylene-NR a R b 、(-C 3-6 Cycloalkylene)-CN and (-C 3-6 Cycloalkylene)-C 1-6 alkyl halide;

[0084] R 14 Each occurrence is independently selected from -L 1 -(C 3-6 Cycloalkyl), -L 1 -(3-10 membered heterocyclic group), -L 1 -(C 6-10 aryl) and -L 1 -(5-14 membered heteroaryl);

[0085] R 41 Selected from C 3-6 Hydrocarbon ring, 3-10 membered heterocycle, C 6-10 Aromatic rings and 5-14 membered heteroaromatic rings;

[0086] Ring C, Ring D, Ring E, Ring X and Ring Z are each independently selected from C 3-6 Hydrocarbon ring, 3-10 membered heterocycle, C 6-10 Aromatic rings and 5-14 membered heteroaromatic rings;

[0087] Ring Y is absent or selected from C 3-6 Hydrocarbon ring, 3-10 membered heterocycle, C6-10 aromatic rings and 5-14 membered heteroaromatic rings; when ring Y is absent, R 24 It does not exist either;

[0088] L 1 Selected from direct bond, -O-, -C(=O)-, -NRC(=O)-, -S-, -S(=O)-, -S(=O)2-, C 1-6 Alkylene and -O-(C 1-6 alkylene)-;

[0089] L 2 Selected from -O-, -C(=O)-, -NRC(=O)-, -S-, -S(=O)-, -S(=O)2-, C 1-6 Alkylene and -O-(C 1-6 alkylene)-;

[0090] R, R a and R b Each occurrence is independently selected from H, C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl and C 6-12 Aralkyl;

[0091] The above alkylene, alkyl, alkenyl, alkynyl, cycloalkylene, cycloalkyl, hydrocarbon ring, heterocyclyl, heterocycle, aryl, aromatic ring, heteroaryl, heteroaromatic ring and aralkyl groups are each optionally substituted with one or more substituents independently selected from the group consisting of halogen, -OH, =O, =CF2, -NH2, -CN, -NO2, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl, C 6-12 Aralkyl, -C(=O)R c 、-OC(=O)R c 、-C(=O)OR c 、-OR c 、-SR c 、-S(=O)R c 、-S(=O)2R c 、-S(=O)2NR c R d 、-NR c R d 、-C(=O)NR c R d 、-NR c -C(=O)R d 、-NRc -C(=O)OR d 、-NR c -S(=O)2-R d 、-NR c -C(=O)-NR c R d 、-C 1-6 Alkylene-OR c 、-C 1-6 Alkylene-NR c R d and -OC 1-6 Alkylene-NR c R d The alkylene, alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and aralkyl groups are further optionally substituted by one or more substituents independently selected from the following: halogen, -OH, =O, -C(=O)O-tert-butyl, -NH2, -CN, -NO2, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl, C 6-12 Aralkyl, -OC 1-6 Alkyl and -C 1-6 Alkylene-OC 1-6 Alkyl; Preferably, the above alkylene, alkyl, alkenyl, alkynyl, cycloalkylene, cycloalkyl, hydrocarbon ring, heterocyclic group, heterocycle, aryl, aromatic ring, heteroaryl, heteroaromatic ring and aralkyl are each optionally substituted by one or more substituents independently selected from the following: halogen, -OH, =O, -NH2, -CN, -NO2, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl, C 6-12 Aralkyl, -C(=O)R c 、-OC(=O)R c 、-C(=O)OR c 、-OR c 、-SR c 、-S(=O)R c 、-S(=O)2R c 、-S(=O)2NR c R d 、-NR c R d 、-C(=O)NR c R d 、-NR c -C(=O)Rd 、-NR c -C(=O)OR d 、-NR c -S(=O)2-R d 、-NR c -C(=O)-NR c R d 、-C 1-6 Alkylene-OR c 、-C 1-6 Alkylene-NR c R d and -OC 1-6 Alkylene-NR c R d The alkylene, alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and aralkyl groups are further optionally substituted by one or more substituents independently selected from the following: halogen, -OH, =O, -C(=O)O-tert-butyl, -NH2, -CN, -NO2, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl, C 6-12 Aralkyl, -OC 1-6 Alkyl and -C 1-6 Alkylene-OC 1-6 alkyl;

[0092] R c and R d Each occurrence is independently selected from H, C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl and C 6-12 Aralkyl, said alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and aralkyl further optionally substituted by one or more substituents independently selected from the following: halogen, -OH, =O, -C(=O)O-tert-butyl, -NH2, -CN, -NO2, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl, C 6- 12 Aralkyl and -C 1-6 Alkylene-OC 1-6 alkyl;

[0093] n, p and q are each independently an integer selected from 1, 2 or 3; and

[0094] m is an integer of 0 or 1.

[0095] In a preferred embodiment, the present disclosure provides a compound or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound, or prodrug thereof, wherein the compound has the structure of the following formula:

[0096] In a preferred embodiment, in the general formula of the present disclosure:

[0097] R 1 for

[0098] R 13 is H, halogen or C 1-6 alkyl;

[0099] R 14 Selected from 3-10 membered heterocyclic group, -(C 1-6 Alkylene)-(3-10 membered heterocyclyl) and -O-(C 1-6 alkylene)-(3-10 membered heterocyclyl), said heterocyclyl being optionally substituted by one or more substituents independently selected from the group consisting of: C 1-6 Alkyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, -C 1-6 Alkylene-C 3-6 Cycloalkyl and -C 1-6 Alkylene-CN; and

[0100] n is an integer of 1 or 2.

[0101] In a preferred embodiment, in the general formula of the present disclosure:

[0102] R 1 for

[0103] R 13 is H, halogen or C 1-6 alkyl;

[0104] R 14 Selected from 3-10 membered heterocyclic group, -(C 1-6 Alkylene)-(3-10 membered heterocyclyl) and -O-(C 1-6 alkylene)-(3-10 membered heterocyclyl), said heterocyclyl being optionally substituted by one or more substituents independently selected from the group consisting of: C 1-6 Alkyl, C 3-6 Cycloalkyl, -C 1-6 Alkylene-C 3-6 Cycloalkyl and -C1-6 Alkylene-CN; and

[0105] n is an integer of 1 or 2.

[0106] In a preferred embodiment, in the general formula of the present disclosure: R 1 for

[0107] In a preferred embodiment, in the general formula of the present disclosure: R 1 for

[0108] In a preferred embodiment, in the general formula of the present disclosure:

[0109] R 1 for

[0110] Ring C is a benzene ring, a 5-6 membered heterocyclic ring or a 5-6 membered heteroaromatic ring;

[0111] Ring D is a benzene ring or a 5-6 membered heteroaromatic ring;

[0112] The above-mentioned benzene ring, heterocyclic ring and heteroaromatic ring are each optionally substituted by one or more substituents independently selected from the following: halogen, -OH, -NH2, -CN, -NO2, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Haloalkyl, -OC 1-6 Alkyl, C 3-6 Preferably, the above-mentioned benzene ring, heterocyclic ring and heteroaromatic ring are each optionally substituted by one or more substituents independently selected from the following: halogen, C 1-6 Alkyl, C 1-6 Haloalkyl, -OC 1-6 Alkyl and C 3-6 Cyclic hydrocarbon group.

[0113] In a preferred embodiment, in the general formula of the present disclosure: R 1 for

[0114] In a preferred embodiment, in the general formula of the present disclosure: R 1 for

[0115] In a preferred embodiment, in the general formula of the present disclosure:

[0116] R 1 -NR 11 R 12 ;

[0117] R 11 and R 12 Each independently selected from H, C 1-6 Alkyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl, C 6-12 Aralkyl, -C 1-6 Alkylene-R a and -C 1-6 Alkylene-NR a R b ;

[0118] R a and R b Each occurrence is independently selected from H, C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl and C 6-12 Aralkyl;

[0119] The alkylene, alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and aralkyl groups are each optionally substituted at each occurrence with one or more substituents independently selected from the group consisting of halogen and C 1-6 alkyl.

[0120] In a preferred embodiment, in the general formula of the present disclosure: R 1 for

[0121] In a preferred embodiment, in the general formula of the present disclosure: R 1 for

[0122] In a preferred embodiment, in the general formula of the present disclosure:

[0123] R 1 for

[0124] Ring E is C 3-6 Hydrocarbon ring, 3-10 membered heterocycle, C 6-10 aromatic ring or 5-14 membered heteroaromatic ring, wherein the hydrocarbon ring and heterocyclic ring are optionally substituted with =CF2;

[0125] R 13 H, halogen, C 1-6Alkyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic, -C(=O)-(C 1-6 alkyl), -O-(C 1-6 alkyl) or -N(C 1-6 alkyl) 2, wherein the alkyl, cycloalkyl and heterocyclic groups are further optionally C 1-6 Alkyl, C 3-6 Substituted with a cycloalkyl or 3-10 membered heterocyclic group;

[0126] n is an integer of 1 or 2.

[0127] In a preferred embodiment, in the general formula of the present disclosure:

[0128] R 1 for

[0129] Ring E is C 3-6 Hydrocarbon ring, 3-10 membered heterocycle, C 6-10 Aromatic ring or 5-14 membered heteroaromatic ring;

[0130] R 13 H, halogen, C 1-6 Alkyl, 3-10 membered heterocyclic group, -C(=O)-(C 1-6 alkyl), -O-(C 1-6 alkyl) or -N(C 1-6 alkyl) 2, wherein the alkyl and heterocyclic groups are further optionally C 1-6 Alkyl or 3-10 membered heterocyclic group substitution;

[0131] n is an integer of 1 or 2.

[0132] In a preferred embodiment, in the general formula of the present disclosure:

[0133] R 1 for

[0134] Ring E is a 3-10 membered heterocyclic ring, C 6-10 Aromatic ring or 5-14 membered heteroaromatic ring;

[0135] R 13 H, halogen, C 1-6 Alkyl, -C(=O)-(C 1-6 alkyl) or -O-(C 1-6 alkyl);

[0136] n is an integer of 1 or 2.

[0137] In a preferred embodiment, in the general formula of the present disclosure: R 1 for

[0138] In a preferred embodiment, in the general formula of the present disclosure: R 1 for

[0139] In a preferred embodiment, in the general formula of the present disclosure: R 1 for

[0140] In a preferred embodiment, in the general formula of the present disclosure:

[0141] When Y does not exist, then

[0142] a) Ring X is a 5-6 membered heterocyclic ring (eg, a pyridone ring) or a 5-membered heteroaromatic ring; or

[0143] b) Ring X is a benzene ring, and at least one R 23 (-C 3-6 Cycloalkylene)-CN or (-C 3-6 Cycloalkylene)-C 1-6 Halogenated alkyl.

[0144] In a preferred embodiment, in the general formula of the present disclosure:

[0145] Ring X is a benzene ring and Ring Y is a 5-6 membered heterocyclic ring or a 5-6 membered heteroaromatic ring.

[0146] In a preferred embodiment, in the general formula of the present disclosure: R 21 、R 22 、R 23 and R 24 Each occurrence is independently selected from H, halogen, -SF5, C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 3-6 Cycloalkyl, -S(=O)2-(C 1-6 alkyl), -S(=O)2-(C 3-6 Cycloalkyl), -P(=O)(C 1-6 Alkyl)2, (-C 3-6 Cycloalkylene)-CN and (-C 3-6 Cycloalkylene)-C 1-6 Halogenated alkyl.

[0147] In a preferred embodiment, in the general formula of the present disclosure: R 2 Selected from:

[0148] In a preferred embodiment, in the general formula of the present disclosure:

[0149] X is a benzene ring or a 5-6 membered heteroaromatic ring (e.g., a pyridine ring); and Y is absent;

[0150] R 23 Each occurrence is independently selected from H, halogen, -SF5, C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 3-6 Cycloalkyl, -S(=O)2-(C 1-6 alkyl), -S(=O)2-(C 3-6 Cycloalkyl), -P(=O)(C 1-6 Alkyl)2, (-C 3-6 Cycloalkylene)-CN and (-C 3-6 Cycloalkylene)-C 1-6 Halogenated alkyl.

[0151] In a preferred embodiment, in the general formula of the present disclosure: R 2 Selected from:

[0152] In a preferred embodiment, in the general formula of the present disclosure: R 3 H or C 1-6 alkyl.

[0153] In a preferred embodiment, in the general formula of the present disclosure: R 3 For ethyl.

[0154] In a preferred embodiment, in the general formula of the present disclosure: Ring Z is a 3-10 membered heterocycle or a benzene ring; preferably a 5-10 membered heterocycle; more preferably a 5-6 membered heterocycle; and

[0155] The heterocyclic ring and the phenyl ring are each optionally substituted at each occurrence by one or more substituents independently selected from the group consisting of halogen and C 1-6 alkyl.

[0156] In a preferred embodiment, in the general formula disclosed herein: Ring Z is

[0157] In a preferred embodiment, in the general formula of the present disclosure: L 2 -C(=O)- or -NRC(=O)-, wherein R is H or C 1-6 alkyl.

[0158] In a preferred embodiment, in the general formula of the present disclosure: L 2 It is -C(=O)-.

[0159] In a preferred embodiment, in the general formula of the present disclosure: R 41 Selected from 3-10 membered heterocyclic ring, C 6-10 aromatic ring and 5-14 membered heteroaromatic ring, wherein the heterocyclic ring, aromatic ring and heteroaromatic ring are each optionally substituted by one or more substituents independently selected from the following: halogen, -OH, -CN, C 1-6 Alkyl, -OC 1-6 Alkyl and -SC 1-6 Alkyl, preferably, the heterocyclic ring, aromatic ring and heteroaromatic ring are at least -OH, -CN or -OC 1-6 Alkyl substitution.

[0160] In a preferred embodiment, in the general formula of the present disclosure: R 41 Selected from 3-10 membered heterocyclic ring, C 6-10 aromatic ring and 5-14 membered heteroaromatic ring, wherein the heterocyclic ring, aromatic ring and heteroaromatic ring are each optionally substituted by one or more substituents independently selected from the following: halogen, -OH, C 1- 6-alkyl, -OC 1-6 Alkyl and -SC 1-6 Alkyl, preferably, the heterocyclic ring, aromatic ring and heteroaromatic ring are at least -OH or -OC 1-6 Alkyl substitution.

[0161] In a preferred embodiment, in the general formula of the present disclosure: -L 2 -R 41 for

[0162] In a preferred embodiment, in the general formula of the present disclosure: -L 2 -R 41 for

[0163] In a preferred embodiment, in the general formula of the present disclosure: R 41 is a 5-6 membered heteroaromatic ring (preferably a pyrazole ring or a pyrimidine ring), which is substituted by at least one -OH group.

[0164] In a preferred embodiment, in the general formula of the present disclosure: R 41 is a 5-membered heteroaromatic ring (preferably a pyrazole ring) substituted by at least one -OH group.

[0165] In a preferred embodiment, in the general formula of the present disclosure: -L 2 -R 41 for

[0166] In a preferred embodiment, in the general formula of the present disclosure: -L 2 -R 41 for

[0167] The present disclosure encompasses technical solutions / compounds obtained by any combination of the various embodiments.

[0168] In a preferred embodiment, the present disclosure provides a compound or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein the compound is selected from:

[0169] In a preferred embodiment, the present disclosure provides a compound or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound, or prodrug thereof, wherein the compound is in a non-zwitterionic form, a zwitterionic form, or a mixture of zwitterionic and non-zwitterionic forms;

[0170] Preferably, the compound is in any one of the following forms or a mixture of any two or three of the following forms:

[0171] Pharmaceutical compositions and methods of treatment

[0172] In some embodiments, the present invention provides a pharmaceutical composition comprising a preventive or therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof and one or more pharmaceutically acceptable carriers. The pharmaceutical composition is preferably a solid preparation, a semi-solid preparation, a liquid preparation or a gaseous preparation. In some embodiments, the pharmaceutical composition may further comprise one or more other therapeutic agents.

[0173] In some embodiments, the present invention provides use of a compound of the present invention, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, or a pharmaceutical composition of the present invention, in the preparation of a medicament for use as a WRN inhibitor.

[0174] In some embodiments, the present invention provides a compound of the present invention, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, or a pharmaceutical composition of the present invention, for use as a WRN inhibitor.

[0175] In some embodiments, the present invention provides a method for preventing or treating cancer (preferably cancer characterized by microsatellite high instability (MSI-H) or mismatch repair deficiency (dMMR)), which comprises administering to a subject in need thereof an effective amount of a compound of the present invention or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, or a pharmaceutical composition of the present invention.

[0176] In some embodiments, the cancer includes colorectal cancer, gastric cancer, endometrial cancer, uterine cancer, adrenocortical cancer, cervical cancer, esophageal cancer, breast cancer, kidney cancer, prostate cancer, and ovarian cancer.

[0177] In the present invention, "pharmaceutically acceptable carrier" refers to a diluent, adjuvant, excipient or vehicle that is administered together with the therapeutic agent and is suitable for contact with the tissues of humans and / or other animals without excessive toxicity, irritation, allergic response or other problems or complications corresponding to a reasonable benefit / risk ratio within the scope of reasonable medical judgment.

[0178] As used herein, unless otherwise indicated, the terms "treat," ...

[0179] As used herein, "subject" includes humans and non-human animals. Exemplary human subjects include human subjects suffering from diseases (e.g., the diseases described herein) (referred to as patients) or normal individuals. "Non-human animals" herein include all vertebrates, such as non-mammals (e.g., birds, amphibians, reptiles) and mammals, such as non-human primates, livestock and / or domesticated animals (e.g., sheep, dogs, cats, cows, pigs, etc.).

[0180] In another embodiment, the pharmaceutical compositions of the present invention may further comprise one or more additional therapeutic or prophylactic agents.

[0181] General synthetic route:

[0182] Route 1

[0183] Route 2

[0184] Route 3

[0185] in:

[0186] P is a protecting group, preferably an amino protecting group, such as Boc;

[0187] Lev is independently a leaving group at each occurrence, such as halogen or OH;

[0188] M is a metal, a borate ester or a boric acid;

[0189] R 0 Each occurrence is independently C 1-6 alkyl.

[0190] In the above routes, the synthesis route is described by taking the compound having an unsubstituted piperazine ring as the Z ring as an example. The above general synthesis route can also be used to synthesize compounds in which the Z ring is a substituted piperazine or a fused piperazine analogue. Example

[0191] The present invention is further described below with reference to examples, but these examples are not intended to limit the scope of the present invention.

[0192] The abbreviations used in this invention have the following meanings:

[0193] Example 1: Synthesis of Compound A1

[0194] 1) Synthesis of intermediate A1-3:

[0195] A1-1 (300 mg, 0.7 mmol), A1-2 (185 mg, 0.84 mmol), 1,4-dioxane (7 mL), water (1 mL), and potassium carbonate (210 mg, 1.54 mmol) were added to a 50 mL single-necked flask. [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (51 mg, 0.07 mmol) was added under nitrogen protection. The mixture was stirred at 100°C under nitrogen protection for 16 hours. The reaction mixture was returned to room temperature and concentrated to dryness under reduced pressure. The mixture was extracted twice with water and dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to afford product A1-3 (90 mg, yield: 24.5%). ESI (m / z) = 523.3 [M+H] + .

[0196] 2) Synthesis of intermediate A1-4:

[0197] Dissolve 2-chloro-4-trifluoromethylaniline (10 g, 51.13 mmol) and potassium carbonate (14.13 g, 102.26 mmol) in dichloromethane (100 mL). Add bromoacetyl bromide (10.32 g, 51.13 mmol) dropwise with stirring at 0°C. Stir and react at 25°C for 1 hour. LCMS indicates the reaction is complete. Pour the reaction solution into water, extract with dichloromethane (250 mL x 2), and concentrate to obtain A1-4 (12 g, yield: 74.2%) as a white solid. m / z [MH] - =314.0. 1 H NMR (400MHz, DMSO-d6) δ10.21(s,1H),8.10(d,J=8.5Hz,1H),7.98(d,J=2.1Hz,1H),7.77(dd,J=8.6,2.1Hz,1H),4.27(s,2H).

[0198] 3) Synthesis of intermediate A1-5:

[0199] A1-3 (90 mg, 0.17 mmol), N,N-dimethylformamide (2 mL), N,N-diisopropylethylamine (66 mg, 0.51 mmol), and A1-4 (70 mg, 0.22 mmol) were added to a 25 mL single-necked flask in sequence and stirred at room temperature for 16 hours. Water and ethyl acetate were added, and the mixture was extracted twice. The organic phases were combined and washed twice with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain the product A1-5 (40 mg, yield: 30.6%). ESI (m / z) = 758.3 [M+H] +

[0200] 4) Synthesis of intermediate A1-6:

[0201] A1-5 (40 mg, 0.053 mmol), dichloromethane (1.5 mL), and trifluoroacetic acid (0.5 mL) were added sequentially to a 25 mL single-necked flask and stirred at room temperature for 1 hour. The reaction solution was concentrated to dryness, and water and dichloromethane were added. The pH was adjusted to 10 with 1% aqueous sodium hydroxide solution. The mixture was separated and extracted twice with DCM. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain product A1-6 (34 mg, yield: 980%), which was used directly in the next step. ESI (m / z) = 658.2 [M+H] + .

[0202] 5) Synthesis of Product A1:

[0203] To a 25 mL single-necked flask, A1-7 (9.8 mg, 0.064 mmol), acetonitrile (1 mL), N-hydroxy-7-azabenzotriazole (8.7 mg, 0.064 mmol), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (15 mg, 0.08 mmol) were added sequentially and stirred at room temperature for 1 hour. A1-6 (35 mg, 0.053 mmol) and N,N-diisopropylethylamine (27 mg, 0.21 mmol) were then added and stirred at room temperature for 1 hour. The reaction mixture was filtered and purified by preparative chromatography (acetonitrile / water = 40%) to afford product A1 (9 mg, yield: 21%). ESI (m / z) = 794.5 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.41(s,1H),8.58(s,1H),8.05(d,J=8.6Hz,1H),7.99–7.90(m ,3H),7.71(dd,J=8.8,2.1Hz,1H),7.03(d,J=9.0Hz,2H),5.37(s,2H),4.53(d,J=12.3Hz ,1H),3.53(d,J=10.0Hz,4H),3.26(d,J=4.8Hz,4H),2.99(d,J=9.7Hz,3H),2.83(d,J=11 .0Hz,1H),2.70–2.60(m,1H),2.46(d,J=8.6Hz,7H),2.24(s,3H),1.20(t,J=7.4Hz,3H).

[0204] Example 2: Synthesis of Compound A2

[0205] 1) Synthesis of intermediate A2-1:

[0206] To a 100 mL single-necked flask were added A1-1 (650 mg, 1.52 mmol), 4-formylphenylboronic acid pinacol ester (460 mg, 1.98 mmol), 1,4-dioxane (13 mL), water (2 mL), and potassium phosphate (810 mg, 3.8 mmol). Methanesulfonic acid (2-dicyclohexylphosphino-2',4',6'-tri-isopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl) palladium(II) (130 mg, 0.15 mmol) was added under nitrogen. The mixture was stirred at 100°C under nitrogen for 16 hours. The reaction mixture was allowed to cool to room temperature, concentrated to dryness under reduced pressure, and extracted twice with water and dichloromethane. The combined organic phases were dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to afford product A2-1 (170 mg, 25% yield). ESI (m / z) = 453.2 [M+H] + .

[0207] 2) Synthesis of intermediate A2-2:

[0208] A2-1 (164 mg, 0.36 mmol), N-methylpiperazine (47 mg, 0.47 mmol), and 1,2-dichloroethane (3 mL) were added to a 25 mL single-necked flask. Acetic acid (24 mg, 0.4 mmol) and sodium triacetoxyborohydride (110 mg, 0.54 mmol) were then added with stirring. The mixture was stirred at room temperature for 3 hours. Methanol was added for dilution, and the pH was adjusted to 8 using ammonia methanol. The mixture was concentrated under reduced pressure and purified by column chromatography to afford product A2-2 (90 mg, yield: 46%). ESI (m / z) = 537.4 [M+H] +

[0209] 3) Synthesis of intermediate A2-3:

[0210] A2-2 (65 mg, 0.12 mmol), N,N-dimethylformamide (1 mL) and N,N-diisopropylethylamine (47 mg, 0.36 mmol) were added to a 25 mL single-necked flask in sequence, and A1-4 (40 mg, 0.13 mmol) was added in two batches and stirred at room temperature for 2 hours. Dichloromethane and water were added to the reaction solution, and the mixture was separated and extracted. The aqueous phase was extracted twice with dichloromethane. The organic phases were combined and washed twice with saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and column chromatography (MeOH / DCM = 7%) gave the product A2-3 (45 mg, yield: 48%). ESI (m / z) = 772.4 [M+H] +

[0211] 4) Synthesis of intermediate A2-4:

[0212] A2-3 (45 mg, 0.058 mmol), dichloromethane (1 mL), and trifluoroacetic acid (0.3 mL) were added sequentially to a 25 mL single-necked flask and stirred at room temperature for 1 hour. The reaction solution was concentrated to dryness, and water and dichloromethane were added. The pH was adjusted to 10 with 1% aqueous sodium hydroxide solution. The mixture was separated and extracted twice with dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give product A2-4 (39 mg, yield: 99%), which was used directly in the next step. ESI (m / z) = 672.3 [M+H] + .

[0213] 5) Synthesis of Product A2:

[0214] To a 25 mL single-necked flask, A1-7 (11 mg, 0.07 mmol), acetonitrile (1 mL), N-hydroxy-7-azabenzotriazole (9.5 mg, 0.07 mmol), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (17 mg, 0.087 mmol) were added sequentially and stirred at room temperature for 1 hour. A2-4 (39 mg, 0.058 mmol) and N,N-diisopropylethylamine (30 mg, 0.23 mmol) were then added and stirred at room temperature for 1 hour. The reaction mixture was filtered and purified by preparative HPLC (acetonitrile / 0.05% aqueous formic acid = 30%) to afford product A2 (14 mg, 30% yield) as the monoformate salt. ESI (m / z) = 808.4 [M+H] + .1H NMR(400MHz,DMSO-d6)δ10.42(s,1H),8.56(s,1H),8.22(s,1H),8.08–8.03( m,3H),7.97(d,J=2.1Hz,1H),7.71(d,J=9.1Hz,1H),7.43(d,J=8.0Hz,2H),5. 39(s,2H),4.53(d,J=12.6Hz,2H),3.52(d,J=11.8Hz,13H),3.02(d,J=9.3Hz, 4H), 2.84 (d, J = 11.4Hz, 2H), 2.44 (s, 3H), 2.15 (s, 3H), 1.20 (d, J = 7.5Hz, 3H).

[0215] Example 3: Synthesis of Compound A31

[0216] 1) Synthesis of intermediate A31-2:

[0217] Compound A31-1 (0.5 g, 1.88 mmol), benzyl alcohol (0.61 g, 5.64 mmol), cuprous iodide (0.36 g, 1.88 mmol), 1,10-phenanthroline (0.12 g, 0.96 mmol), and cesium carbonate (3.06 g, 9.40 mmol) were dissolved in toluene (5 mL) and stirred at 100°C for 16 hours. LCMS showed the reaction was complete, and the reaction solution was concentrated and purified by column chromatography to give A31-2 (0.05 g, yield: 10%) as a white solid. ESI m / z [M+H] + =247.2.

[0218] 2) Synthesis of intermediate A31-3:

[0219] Compound A31-2 (50 mg, 0.20 mmol) and sodium hydroxide (24 mg, 0.60 mmol) were dissolved in a solution of methanol (0.5 mL) and water (0.5 mL) and stirred at 45°C for 4 hours. LCMS showed the reaction was complete. The pH was adjusted to 4 and the reaction solution was concentrated to give A31-3 (0.04 g, yield: 85%) as a white solid. ESI m / z [M+H] + =233.1.

[0220] 3) Synthesis of intermediate A31-4:

[0221] Compound A31-3 (0.04 g, 0.17 mmol), A4-3 (0.12 g, 0.20 mmol), HATU (0.10 g, 0.26 mmol), and DIEA (0.07 g, 0.51 mmol) were dissolved in DMF (1 mL) and stirred at 25° C. for 16 hours. LCMS showed the reaction was complete, and the reaction solution was used directly in the next step.

[0222] 4) Synthesis of product A31:

[0223] Trifluoroacetic acid (1 mL) was added to the reaction mixture and stirred at 80°C for 16 hours. LCMS indicated the reaction was complete. The reaction mixture was poured into water and extracted with dichloromethane (10 mL x 2). The mixture was concentrated and purified by reverse phase chromatography to afford A31 as a white solid (3.8 mg, yield: 3%, purity: 95.57%). ESI m / z [MH] - =688.4. 1H NMR (400MHz, DMSO-d6) δ10.40(s,1H),9.33(s,1H),8.08(d,J=8.6Hz,1H),7.99(s,1H),7.75(d,J=8.3Hz,1H),7.41(s,1H),6.86(s,1H), 5.35(s,2H),5.02(s,1H),4.57(s,1H),4.28(s,2H),4.14(q,J=5.2Hz,2H),3.82(d ,J=6.1Hz,6H),3.03(d,J=8.1Hz,2H),2.80(s,1H),2.10–1.96(m,4H),1.36(s,3H).

[0224] Example 4: Synthesis of Compound A4

[0225] 1) Synthesis of intermediate A4-2:

[0226] Compound A4-1 (0.8 g, 1.86 mmol), compound A1-4 (0.65 g, 2.05 mmol), and N,N-diisopropylethylamine (0.72 g, 5.58 mmol) were dissolved in N,N-dimethylformamide (6 mL) and stirred at 45°C for 5 hours. LCMS showed the reaction was complete. The reaction solution was poured into water, extracted with dichloromethane (10 mL x 2), and concentrated to obtain a brown solid A4-2 (0.8 g, yield: 64.6%). m / z [M+H] + =666.1.

[0227] 2) Synthesis of intermediate A4-3:

[0228] Compound A4-2 (0.15 g, 0.23 mmol) was dissolved in dichloromethane (2 mL) and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at 25°C for 2 hours. LCMS showed that the reaction was complete. The reaction mixture was concentrated to obtain a brown oil A4-3 (0.12 g, yield: 94.2%). m / z [M+H] + =566.2.

[0229] 3) Synthesis of product A4:

[0230] Compound A4-3 (120 mg, 0.21 mmol), N-hydroxy-7-azabenzotriazole (31 mg, 0.23 mmol), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (60 mg, 0.32 mmol) were dissolved in acetonitrile (3 mL) and stirred at 25°C for 1 hour. Compound A4-4 (65 mg, 0.32 mmol) and N,N-diisopropylethylamine (81 mg, 0.63 mmol) were then added to the reaction solution, and the reaction solution was stirred at 25°C for another 1 hour. LCMS showed that the reaction was complete, and the reaction solution was filtered to obtain a filtrate, which was purified by reverse phase column chromatography (0.05% aqueous formic acid / acetonitrile) to obtain a white product A4 (21 mg, yield: 12.4%, purity: 94.5%). m / z[M+H] + =754.3. 1 H NMR (400MHz, DMSO-d6) δ10.41(s,1H),8.97(d,J=6.9Hz,1H),8.16(t,J=7.9Hz,1H),8.09(d,J=8.6Hz,1H), 7.99(d,J=2.0Hz,1H),7.74(dd,J=8.8,2.1Hz,1H),7.45–7.35(m,2H),6.86(s,1H),5.35(s,2H),4.55(d,J= 12.4Hz,1H),4.28(q,J=3.1Hz,2H),3.84(q,J=5.4Hz,3H),3.55–3.48(m,3H),3.28–3.15(m,3H),3.02(d,J =8.7Hz,2H),2.87(t,J=12.3Hz,1H),2.78(d,J=10.5Hz,1H),2.62(d,J=11.0Hz,1H),1.22(t,J=7.4Hz,3H).

[0231] Example 5: Synthesis of Compound A5

[0232] A5-1 (50 mg, 0.088 mmol) and DMF (1 mL) were added to a reaction flask. HOBT (20 mg, 0.15 mmol) and EDCI (40 mg, 0.21 mmol) were added sequentially with stirring. The mixture was stirred at room temperature for 1 hour (referred to as Solution 1). In a separate reaction flask, A4-3 (50 mg, 0.088 mmol), DIPEA (50 mg, 0.39 mmol), and acetonitrile (1 mL) were added (referred to as Solution 2). Solution 2 was added to Solution 1 and the mixture was reacted at 25°C for 2 hours. The reaction was monitored by LCMS and reverse phase preparative separation afforded A5 (4.4 mg, 7%) as a white solid. 1H NMR (400MHz, DMSO-d6) δ10.37(s,1H),8.06(d,J=8.6Hz,1H),7.97(s,1H),7.72(d,J=8.8Hz,1H), 6.83(s,1H),5.31(s,2H),4.47(d,J=12.0Hz,1H),4.25(s,2H),3.80(t,J=5.6Hz,2H),3.68(s,4H ),3.48(s,1H),3.17(d,J=11.7Hz,1H),2.98(s,2H),2.76(d,J=19.2Hz,4H),2.60(d,J=10.3Hz,1 H),1.83(d,J=5.6Hz,2H),1.77–1.73(m,2H),1.23(s,2H),1.19(t,J=7.5Hz,3H).LCMS(ESI)[M+1] + =758.3.

[0233] Example 6: Synthesis of Compound A6

[0234] To a reaction flask, add A4-3 (50 mg, 0.088 mmol), A6-1 (20 mg, 0.13 mmol), HATU (50 mg, 0.13 mmol), DIPEA (50 mg, 0.39 mmol), and DMF (1 mL). Incubate at 25°C for 1 hour. LCMS monitored the reaction for completion. Reverse-phase preparative chromatography isolated A6 (22 mg, 35%) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ10.37(s,1H),8.05(d,J=8.6Hz,1H),7.96(d,J=2.1Hz,1H),7.81(dd,J=6.8,2.1Hz,1H),7.71(dd, J=8.8,2.1Hz,1H),7.51(dd,J=6.9,2.0Hz,1H),6.82(dd,J=3.2,1.6Hz,1H),6.29(t,J=6.8Hz,1H),5.31(s,2H),4.48(d,J =12.5Hz,1H),4.25(q,J=2.9Hz,2H),3.80(t,J=5.5Hz,2H),3.47(s,3H),3.42(d,J=8.9Hz,4H),3.29–3.18(m,2H),2.98(q ,J=7.3Hz,2H),2.93–2.85(m,1H),2.76(d,J=11.6Hz,1H),2.62(d,J=10.9Hz,1H),1.18(t,J=7.5Hz,3H).LCMS(ESI)[M+1]+ =701.3.

[0235] Example 7: Synthesis of Compound A9

[0236] 1) Synthesis of intermediate A9-2:

[0237] Compound A9-1 (0.2 g, 1.16 mmol) and potassium carbonate (0.32 g, 2.32 mmol) were dissolved in dichloromethane (2 mL). Bromoacetyl bromide (0.26 g, 1.28 mmol) was added dropwise with stirring at 0°C. The mixture was stirred at 25°C for 1 hour. LCMS indicated the reaction was complete. The reaction solution was poured into water, extracted with dichloromethane (5 mL x 2), and concentrated to afford A9-2 (0.28 g, yield: 82.4%) as a brown solid. m / z [MH] - =292.0.

[0238] 2) Synthesis of intermediate A9-3:

[0239] Compound A4-1 (0.1 g, 0.23 mmol), compound A9-2 (74 mg, 0.25 mmol), and N,N-diisopropylethylamine (89 mg, 0.69 mmol) were dissolved in N,N-dimethylformamide (4 mL) and stirred at 45°C for 5 hours. LCMS showed the reaction was complete. The reaction solution was poured into water, extracted with ethyl acetate (5 mL*2), and concentrated to obtain a brown solid A9-3 (0.14 g, yield: 93.6%). m / z [M+H] + =644.2.

[0240] 3) Synthesis of intermediate A9-4:

[0241] Compound A9-3 (0.14 g, 0.22 mmol) was dissolved in dichloromethane (42 mL) and trifluoroacetic acid (2 mL) was added. The reaction mixture was stirred at 25°C for 1 hour. LCMS showed the reaction was complete. The reaction mixture was poured into water, extracted with dichloromethane (5 mL x 2), and concentrated to obtain a brown oil, A9-4 (0.12 g, yield: 99.5%). m / z [M+H] + =544.2.

[0242] 4) Synthesis of product A9:

[0243] Compound A1-7 (41 mg, 0.26 mmol), N-hydroxy-7-azabenzotriazole (30 mg, 0.22 mmol), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (63 mg, 0.33 mmol) were dissolved in acetonitrile (3 mL) and stirred at 25°C for 1 hour. Compound A9-4 (120 mg, 0.22 mmol) and N,N-diisopropylethylamine (85 mg, 0.66 mmol) were then added sequentially to the reaction solution, which was stirred at 25°C for another 1 hour. LCMS indicated the reaction was complete, and the reaction solution was filtered to obtain the filtrate, which was purified by reverse phase column chromatography (0.05% aqueous formic acid / acetonitrile) to obtain the white product A9 (45 mg, yield: 27.74%, purity: 92.49%). m / z [M+H] + =680.3. 1 H NMR (400MHz, DMSO-d6) δ10.78(s,1H),8.60(d,J=2.8Hz,1H),7.74(d,J=2.1Hz,1H),7.41(dd,J=8.8,2. 3Hz,1H),7.29(dd,J=8.8,2.2Hz,1H),6.83(t,J=2.4Hz,1H),5.19(s,2H),4.55(d,J=12.6Hz,1H),4.31 –4.23(m,2H),3.82(t,J=5.3Hz,2H),3.58–3.49(m,4H),3.30–3.26(m,1H),3.01(ddt,J=17.7,14.3,6. 3Hz, 3H), 2.83 (dd, J=15.7, 9.3Hz, 2H), 2.68 (t, J=9.3Hz, 1H), 2.47 (d, J=2.1Hz, 3H), 1.23–1.15 (m, 3H).

[0244] Example 8: Synthesis of Compound A10

[0245] Referring to the synthesis method of Experimental Example 7, Instead of A9-1, compound A10 was synthesized. m / z[M+H] + =665.4. 1H NMR(400MHz,DMSO-d6)δ10.60(s,1H),8.56(s,1H),7.62–7.51(m,2H),7.28(dd,J=9.1,2.5Hz,2 H),6.83–6.74(m,1H),5.13(s,2H),4.50(d,J=12.5Hz,1H),4.22(q,J=2.8Hz,2H),3.78(t,J=5.4 Hz,3H),3.44(s,4H),3.24(t,J=12.1Hz,1H),3.04–2.88(m,3H),2.81(d,J=10.7Hz,1H),2.63(d, J=10.7Hz,1H),2.43(s,3H),1.69(q,J=4.9Hz,2H),1.42(q,J=5.1Hz,2H),1.15(t,J=7.5Hz,3H).

[0246] Example 9: Synthesis of Compound A11

[0247] Referring to the synthesis method of Experimental Example 7, Instead of A9-1, compound A11 was synthesized. m / z[M+H] + =679.4. 1 H NMR (400MHz, DMSO-d6) δ10.62(s,1H),10.26(s,1H),8.60(s,1H),7.70–7.57(m,2H),7.51–7.37(m,2H),6.83( s,1H),5.19(s,2H),4.55(d,J=12.6Hz,1H),4.27(d,J=3.1Hz,2H),3.82(t,J=5.3Hz,2H),3.52(t,J=11.3Hz,3 H),3.28(s,1H),3.00(d,J=11.0Hz,3H),2.85(d,J=11.0Hz,1H),2.73(ddt,J=12.0,7.6,3.8Hz,3H),2.67–2.5 4(m,4H),2.47(d,J=2.0Hz,3H),2.38–2.22(m,1H),2.02(ddd,J=13.8,8.9,4.5Hz,1H),1.20(t,J=7.5Hz,3H).

[0248] Example 10: Synthesis of Compound A12

[0249] Referring to the synthesis method of Experimental Example 7, Instead of A9-1, compound A12 was synthesized. m / z[M+H]+ =714.3. 1 H NMR (400MHz, DMSO-d6) δ10.31 (s, 1H), 8.60 (s, 1H), 7.79 (d, J = 22.3Hz, 2H) ,6.87(s,1H),5.26(s,2H),4.55(d,J=12.5Hz,1H),4.29(s,2H),3.84(t,J =5.6Hz,2H),3.52(d,J=11.5Hz,3H),3.01(q,J=10.2,7.4Hz,4H),2.84(d, J=11.2Hz,1H),2.66(d,J=10.9Hz,1H),2.47(s,5H),1.22(t,J=7.5Hz,4H).

[0250] Example 11: Synthesis of Compound A18

[0251] Referring to the synthesis method of Experimental Example 7, Instead of A9-1, compound A18 was synthesized. m / z[M+H] + =691.3. 1 H NMR (400MHz, DMSO-d6) δ10.37(s,1H),9.49(s,1H),8.57(s,1H),8.12(d,J=8.7Hz,1H),7.74(d, J=8.7Hz,1H),6.86(d,J=1.7Hz,1H),5.28(s,2H),4.52(d,J=12.5Hz,1H),4.27(q,J=2.8Hz,2H) ,3.82(q,J=5.5,4.7Hz,2H),3.57–3.44(m,3H),3.29–3.19(m,2H),3.01(p,J=8.2Hz,3H),2.82( d,J=10.3Hz,1H),2.64(d,J=11.2Hz,1H),2.53(d,J=3.3Hz,1H),2.44(s,3H),1.29–1.18(m,3H).

[0252] Example 12: Synthesis of Compound A22

[0253] Referring to the synthesis method of Experimental Example 7, Instead of A9-1, compound A22 was synthesized. m / z[M+H] + =708.4. 1H NMR (400MHz, DMSO-d6) δ10.61(s,1H),10.22(s,1H),8.59(s,1H),7.67–7.57(m,2H),7.43(d,J=8.3 Hz,2H),6.85–6.78(m,1H),5.18(s,2H),4.55(d,J=12.4Hz,1H),4.26(q,J=2.8Hz,2H),3.81(t,J=5 .4Hz,2H),3.60–3.46(m,3H),3.29(d,J=15.0Hz,3H),3.00(dt,J=13.1,6.0Hz,3H),2.84(d,J=11.7 Hz,1H),2.66(d,J=11.0Hz,1H),2.47(s,3H),1.37–1.30(m,2H),1.19(t,J=7.4Hz,3H),1.09(s,2H).

[0254] Example 13: Synthesis of Compound A17

[0255] Referring to the synthesis method of Experimental Example 7, Instead of A9-1, compound A17 was synthesized. m / z[M+H] + =685.3. 1 H NMR (400MHz, DMSO-d6) δ10.52(s,1H),9.04(dd,J=4.2,1.7Hz,1H),8.60(d,J=2.8Hz,1H),8.46(dt,J=8.3,2.1H z,1H),8.01(q,J=9.0Hz,2H),7.64(dd,J=8.2,4.2Hz,1H),6.94–6.85(m,1H),5.38(s,2H),4.56(d,J=12.3Hz,1 H),4.29(q,J=2.8Hz,2H),3.83(q,J=4.3,3.3Hz,2H),3.61–3.48(m,3H),3.30(s,2H),3.04(dd,J=16.9,9.2Hz, 3H), 2.86 (d, J = 11.6Hz, 1H), 2.68 (d, J = 10.5Hz, 1H), 2.55 (s, 1H), 2.47 (d, J = 2.0Hz, 3H), 1.25 (t, J = 7.4Hz, 3H).

[0256] Example 14: Synthesis of Compound A36

[0257] 1) Synthesis of intermediate A36-1:

[0258] A1-1 (1 g, 2.34 mmol), A1-4 (0.81 g, 2.57 mmol), and N,N-diisopropylethylamine (0.91 g, 7.02 mmol) were dissolved in acetonitrile (10 mL) and stirred at 45°C for 4 hours. LCMS showed the reaction was complete, and the reaction solution was filtered to obtain A36-1 (0.78 g, yield: 50.3%) as a white solid. ESI m / z [M+H] + =662.1.

[0259] 2) Synthesis of intermediate A36-2:

[0260] A36-1 (0.2 g, 0.3 mmol), dimethylamine hydrochloride (0.12 g, 1.5 mmol), and N,N-diisopropylethylamine (0.12 g, 0.9 mmol) were dissolved in acetonitrile (3 mL) and stirred at 80°C for 16 hours. LCMS showed the reaction was complete, and the reaction solution was concentrated to give the crude product A36-2 as a yellow solid (0.2 g, yield: 95.1%). ESI m / z [M+H] + =627.3.

[0261] 3) Synthesis of intermediate A36-3:

[0262] A36-2 (0.21 g, 0.33 mmol) was dissolved in dichloromethane (2 mL) and trifluoroacetic acid (2 mL) was added. The reaction mixture was stirred at 25°C for 2 hours. LCMS showed that the reaction was complete. The reaction mixture was concentrated to give A36-3 (0.15 g, yield: 85%) as a yellow oil. ESI m / z [M+H] + =527.2.

[0263] 4) Synthesis of product A36:

[0264] Compound A1-7 (52 mg, 0.34 mmol), N-hydroxy-7-azabenzotriazole (42 mg, 0.31 mmol), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (81 mg, 0.42 mmol) were dissolved in N,N-dimethylformamide (0.5 mL) and stirred at 25°C for 1 hour. A36-3 (150 mg, 0.28 mmol) and N,N-diisopropylethylamine (110 mg, 0.84 mmol) were then added to the reaction solution, and the reaction solution was stirred at 25°C for another 2 hours. LCMS showed that the reaction was complete, and the reaction solution was filtered to obtain the filtrate, which was purified by reverse phase column chromatography (0.05% aqueous formic acid / acetonitrile) to obtain the white product A36 (17 mg, yield: 8.8%, purity: 97.9%). ESI m / z [M+H]+ =663.3. 1 H NMR (400MHz, DMSO-d6) δ10.34(s,1H),8.55(s,1H),8.04(d,J=8.6Hz,1H),7.95( d,J=2.0Hz,1H),7.72(dd,J=8.8,2.1Hz,1H),5.20(s,2H),4.50(d,J=12.5Hz,1H ),3.49(d,J=12.4Hz,4H),3.29–3.17(m,2H),2.95(s,6H),2.93–2.88(m,1H),2. 77(d,J=11.1Hz,1H),2.59(d,J=10.9Hz,1H),2.43(s,3H),1.15(t,J=7.4Hz,3H).

[0265] Example 15: Synthesis of Compound A26

[0266] 1) Synthesis of intermediate A26-2:

[0267] A36-1 (100 mg, 0.15 mmol), A26-1 (55 mg, 0.17 mmol) and potassium carbonate (62 mg, 0.45 mmol) were dissolved in a mixed solution of 1,4-dioxane (3 mL) and water (0.4 mL). The air was replaced three times with nitrogen, and then 1,1'-bis(diphenylphosphino)ferrocenepalladium (II) chloride was added and stirred at 100 ° C for 16 hours. LCMS showed that the reaction was complete. The materials were added twice in parallel and combined for post-processing. The liquid was separated, the organic phase was spin-dried, and purified by silica gel column chromatography (ethyl acetate / petroleum ether: 50-80%) to give compound A26-2 (140 mg, average yield: 84%). ESI m / z[M+H] + =789.6.

[0268] 2) Synthesis of intermediate A26-3:

[0269] Compound A26-2 (100 mg, 0.13 mmol) was dissolved in dichloromethane (2 mL) and trifluoroacetic acid (0.5 mL) was added. The reaction mixture was stirred at 25°C for 2 hours. LCMS showed that the reaction was complete. The reaction mixture was concentrated to obtain a brown oil A26-3 (50 mg, yield: 57.3%). ESI m / z [M+H] + =689.6.

[0270] 3) Synthesis of product A26:

[0271] Compound A1-7 (14 mg, 0.09 mmol), N-hydroxy-7-azabenzotriazole (21 mg, 0.11 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (12 mg, 0.09 mmol) were dissolved in N,N-dimethylformamide (1.5 mL) and stirred at 25°C for 1 hour. Compound A26-3 (50 mg, 0.07 mmol) and N,N-diisopropylethylamine (28 mg, 0.22 mmol) were then added to the reaction solution in sequence, and the reaction solution was stirred at 25°C for another 1 hour. LCMS showed that the reaction was complete, and the reaction solution was directly purified by reverse phase column chromatography to obtain a white product A26 (5.5 mg, yield: 9.2%, purity: 93.7%). ESI m / z[M+H] + =825.4. 1 H NMR (400MHz, DMSO-d6) δ10.41 (s, 1H), 8.57 (s, 1H), 8.04 (dd, J = 11.1, 8.5Hz, 4H), 7. 96(d,J=2.0Hz,1H),7.74–7.68(m,1H),7.08(d,J=8.5Hz,2H),5.38(s,2H),4.53(d,J =12.3Hz,1H),4.15(t,J=5.7Hz,2H),3.58(t,J=4.7Hz,4H),3.52(d,J=11.4Hz,4H),3 .07–2.94(m,3H),2.84(d,J=11.2Hz,1H),2.74–2.65(m,3H),2.49–2.46(m,4H),2.44 (s,3H),1.21(s,3H).

[0272] Example 16: Synthesis of Compound A27

[0273] Reference Example 15: Synthesis of Compound A26, using Substituting A26-1, compound A27 was synthesized. ESI m / z[M+H] + =809.5. 1H NMR (400MHz, DMSO-d6) δ10.44(s,1H),8.61(s,1H),8.23–7.94(m,5H),7.74(dd,J= 8.8,2.1Hz,1H),7.11(d,J=8.6Hz,2H),5.41(s,2H),4.56(d,J=12.3Hz,1H),4.19( t,J=5.7Hz,2H),3.28(d,J=12.4Hz,4H),3.04(q,J=9.1,8.0Hz,4H),2.92(t,J=5.7 Hz,3H),2.65(d,J=5.9Hz,4H),2.48(s,3H),1.79–1.71(m,4H),1.27–1.21(m,3H).

[0274] Example 17: Synthesis of Compound A85

[0275] A85-1 (30 mg, 0.22 mmol), N-hydroxy-7-azabenzotriazole (37 mg, 0.27 mmol), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (62 mg, 0.32 mmol) were dissolved in N,N-dimethylformamide (0.5 mL) and stirred at 25°C for 1 hour. Compound A4-3 (100 mg, 0.18 mmol) and N,N-diisopropylethylamine (70 mg, 0.54 mmol) were then added to the reaction solution, and the reaction solution was stirred at 25°C for another 1 hour. LCMS showed that the reaction was complete, and the reaction solution was filtered to obtain a filtrate, which was purified by reverse phase column chromatography (0.05% aqueous formic acid / acetonitrile) to obtain the white product A85 (22 mg, yield: 17.6%, purity: 97%). ESI m / z [M+H] + =687.3. 1H NMR (400MHz, DMSO-d6) δ10.40(s,1H),8.08(d,J=8.6Hz,1H),7.99(d,J=2.1Hz,1H),7.87(s ,1H),7.81–7.65(m,2H),6.93–6.81(m,1H),6.22(s,1H),5.34(s,2H),4.51(d,J=12.6Hz,1 H),4.28(p,J=3.2Hz,2H),3.83(t,J=5.5Hz,2H),3.45(s,4H),3.26(s,1H),3.01(q,J=7.4H z,2H),2.95–2.73(m,3H),2.66(d,J=10.5Hz,1H),2.15–1.73(m,1H),1.22(t,J=7.4Hz,3H).

[0276] Example 18: Synthesis of Compound A79

[0277] 1) Synthesis of intermediate A79-2:

[0278] A36-1 (150 mg, 0.23 mmol), potassium carbonate (95 mg, 0.69 mmol) and 2-methyl-2H-indazole-5-boronic acid (45 mg, 0.25 mmol) were dissolved in a mixed solution of 1,4-dioxane (1 mL) and water (0.1 mL). After replacing the gas three times, 1,1-bis(diphenylphosphino)ferrocenepalladium(II) chloride (17 mg, 0.03 mmol) was added and stirred at 80°C for 16 hours. LCMS showed that the reaction was complete. The reaction solution was poured into water and extracted with dichloromethane (10 mL*2). The organic phases were combined, concentrated, and purified by silica gel column chromatography (ethyl acetate / petroleum ether) to obtain A79-2 (0.1 g, yield: 61.9%) as a white solid. ESI m / z[M+H] + =714.3.

[0279] 2) Synthesis of intermediate A79-3:

[0280] A79-2 (100 mg, 0.14 mmol) was dissolved in dichloromethane (1 mL) and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at 25°C for 2 hours. LCMS showed that the reaction was complete. The reaction mixture was concentrated to give A79-3 (85 mg, yield: 98%) as a yellow oil. ESI m / z [M+H] + =614.1.

[0281] 3) Synthesis of product A79:

[0282] Compound A1-7 (28 mg, 0.18 mmol), N-hydroxy-7-azabenzotriazole (21 mg, 0.15 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (40 mg, 0.21 mmol) were dissolved in N,N-dimethylformamide (3 mL) and stirred at 25°C for 1 hour. Compound A79-3 (86 mg, 0.14 mmol) and N,N-diisopropylethylamine (54 mg, 0.42 mmol) were then added to the reaction solution, and the reaction solution was stirred at 25°C for another 2 hours. LCMS showed that the reaction was complete, and the reaction solution was filtered to obtain a filtrate, which was subjected to reverse phase column chromatography (0.05% aqueous formic acid / acetonitrile) to obtain a white product A79 (32 mg, yield: 30.4%, purity: 99.8%). ESI m / z [MH] - =748.4. 1 H NMR (400MHz, DMSO-d6) δ10.43(s,1H),8.58(s,1H),8.53–8.47(m,2H),8.07(d,J= 8.6Hz,1H),8.00–7.94(m,2H),7.74–7.66(m,2H),5.41(s,2H),4.54(d,J=12.0Hz ,1H),4.19(s,3H),3.52(t,J=10.6Hz,3H),3.27(s,1H),3.02(d,J=9.2Hz,3H),2. 85(d,J=11.2Hz,1H),2.67(d,J=10.6Hz,1H),2.45(s,3H),1.21(t,J=5.6Hz,3H).

[0283] Example 19: Synthesis of Compound A76

[0284] Reference Example 18 Synthesis of Compound A79, using Substituting A79-1, compound A76 was synthesized. ESI m / z [MH] - =748.5. 1H NMR(400MHz,DMSO-d6)δ10.46(s,1H),8.61(s,1H),8.56(d,J=1.4Hz,1H),8.24–8.15(m,2H), 8.10(d,J=8.6Hz,1H),8.00(d,J=2.0Hz,1H),7.79(d,J=8.8Hz,1H),7.74(dd,J=8.7,2.2Hz,1 H),5.44(s,2H),4.57(d,J=12.4Hz,1H),4.11(s,3H),3.57(q,J=10.2,8.6Hz,3H),3.04(t,J= 11.0Hz,3H),2.88(d,J=11.3Hz,1H),2.70(d,J=10.5Hz,1H),2.48(s,3H),1.28–1.24(m,3H).

[0285] Example 20: Synthesis of Compound A78

[0286] Reference Example 18 Synthesis of Compound A79, using Substituting A79-1, compound A78 was synthesized. ESI m / z [MH] - =735.4. 1 H NMR (400MHz, DMSO-d6) δ10.44(s,1H),9.41–9.34(m,2H),8.61(s,1H),8.11(d,J=8. 6Hz,1H),8.03–7.99(m,2H),7.94(d,J=9.6Hz,1H),7.74(dd,J=8.8,2.1Hz,1H),5.4 3(s,2H),4.57(d,J=12.1Hz,1H),3.55(d,J=11.6Hz,3H),3.30(s,1H),3.10–3.00(m ,3H),2.92–2.85(m,1H),2.71(d,J=10.7Hz,1H),2.47(s,3H),1.24(d,J=7.7Hz,3H).

[0287] Example 21: Synthesis of Compound A19

[0288] 1) Synthesis of intermediate A19-2:

[0289] A19-1 (1 g, 5.87 mmol) and NBS (1.23 g, 6.94 mmol) were dissolved in dichloromethane (10 mL) and stirred at room temperature for 1 hour. LCMS showed the reaction was complete, and saturated aqueous sodium thiosulfate was added to quench the reaction. The organic phase was separated and dried by spin drying. Compound A19-2 (1.1 g, mixture, yield: 75.6%) was purified by silica gel column chromatography (ethyl acetate / petroleum ether: 50-80%). ESI m / z [M+H] + =252.0.

[0290] 2) Synthesis of intermediate A19-3:

[0291] Compound A19-2 (1.1 g, 4.36 mmol), cupric chloride (1.17 g, 8.72 mmol), and tert-butyl nitrite (1.17 g, 8.72 mmol) were dissolved in acetonitrile (10 mL). The solution was stirred at 50 ° C for 30 minutes and then cooled to room temperature. The reaction mixture was quenched with 5% hydrochloric acid solution, diluted with deionized water, and extracted with ethyl acetate (2X 50 mL). Vacuum concentration and purification by silica gel column chromatography (ethyl acetate / petroleum ether: 50-80%) gave compound A19-3 (1 g, mixture, yield: 84.4%), which was directly carried out to the next step.

[0292] 3) Synthesis of intermediate A19-4:

[0293] A19-3 (1 g, 3.68 mmol), tert-butyl carbamate (0.65 g, 5.52 mmol) and cesium carbonate (4.8 g, 14.7 mmol) were dissolved in a 1,4-dioxane (10 mL) solution. The air was replaced three times with nitrogen. 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (0.64 g, 1.1 mmol) and tris(dibenzylideneacetone)dipalladium (0.34 g, 0.37 mmol) were then added and stirred at 110 ° C for 6 hours. LCMS showed that the reaction was complete, the organic phase was separated and dried, and purified by silica gel column chromatography (ethyl acetate / petroleum ether: 50-80%) to give compound A19-4 (450 mg, mixture, yield: 39.7%). ESI m / z [M+H] + =308.1.

[0294] 4) Synthesis of intermediate A19-5:

[0295] A19-4 (450 mg, 1.46 mmol) was dissolved in dichloromethane (3 mL) and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at 25°C for 2 hours. LCMS showed that the reaction was complete. The reaction mixture was concentrated to give A19-5 (300 mg, mixture, yield: 98.8%) as a brown oil. ESI m / z [M+H] + =208.1.

[0296] 5) Synthesis of intermediate A19-6:

[0297] Compound A19-5 (0.3 g, 1.45 mmol) and potassium carbonate (0.6 g, 4.35 mmol) were dissolved in dichloromethane (2 mL). Bromoacetyl bromide (0.44 g, 2.17 mmol) was added dropwise with stirring at 0°C. The mixture was stirred at 25°C for 1 hour. LCMS indicated the reaction was complete. The reaction solution was poured into water, extracted with dichloromethane (50 mL x 2), and concentrated to afford A19-6 (0.3 g, mixture, yield: 63.2%) as a yellow solid. Proceed directly to the next step.

[0298] 6) Synthesis of intermediate A19-7:

[0299] Compound A4-1 (0.13 g, 0.3 mmol), A19-6 (0.1 g, 0.3 mmol), and N,N-diisopropylethylamine (0.12 g, 0.9 mmol) were dissolved in N,N-dimethylformamide (1 mL) and stirred at 45°C for 5 hours. LCMS showed the reaction was complete. The reaction solution was poured into water, extracted with ethyl acetate (100 mL*2), and concentrated to obtain a brown solid A19-7 (0.15 g, mixture, yield: 73.3%). ESI m / z [M+H] + =678.3.

[0300] 7) Synthesis of intermediate A19-8:

[0301] Compound A19-7 (0.15 g, 0.22 mmol) was dissolved in dichloromethane (2 mL) and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at 25°C for 1 hour. LCMS showed that the reaction was complete. The mixture was concentrated to give A19-8 (0.1 g, mixture, yield: 78.2%) as a brown oil. ESI m / z [M+H] + =578.1.

[0302] 8) Synthesis of product A19:

[0303] Compound A1-7 (31 mg, 0.2 mmol), N-hydroxy-7-azabenzotriazole (49 mg, 0.26 mmol), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (28 mg, 0.20 mmol) were dissolved in N,N-dimethylformamide (2 mL) and stirred at 25°C for 1 hour. Subsequently, compound A19-8 (100 mg, 0.17 mmol) and N,N-diisopropylethylamine (66 mg, 0.51 mmol) were added to the reaction solution in sequence, and the reaction solution was stirred at 25°C for another 4 hours. LCMS showed the reaction was complete. The reaction solution was filtered to obtain the filtrate, which was purified by reverse phase column chromatography (0.05% aqueous formic acid / acetonitrile) to give white products A19 (4.5 mg, yield: 3.2%, purity: 93.1%) and A19' (4.5 mg, yield: 3.2%).

[0304] A19:ESI m / z[M+H] + =714.3. 1 H NMR (400MHz, DMSO-d6) δ10.36 (s, 1H), 8.54 (s, 1H), 7.44 (d, J = 2.2Hz, 2H), 6. 88(d,J=2.8Hz,1H),5.24(s,2H),4.54(d,J=12.5Hz,1H),4.29(q,J=2.8Hz,2H ),3.84(t,J=5.4Hz,2H),3.57–3.49(m,4H),3.01(q,J=8.6,8.1Hz,4H),2.83( d,J=11.4Hz,2H),2.65(d,J=11.0Hz,1H),2.45(s,3H),1.22(t,J=7.5Hz,3H).

[0305] A19':ESI m / z[M+H] + =714.3. 1H NMR (400MHz, DMSO-d6) δ10.85(s,1H),8.59(s,1H),7.60(d,J=9.2Hz,1H),7.34(d,J= 9.2Hz,1H),6.93–6.81(m,1H),5.25(s,2H),4.55(d,J=12.6Hz,1H),4.27(q,J=2.9Hz ,2H),3.83(t,J=5.4Hz,2H),3.53(d,J=12.1Hz,3H),3.28(s,3H),3.00(t,J=7.7Hz,3 H), 2.84 (d, J = 11.4Hz, 1H), 2.67 (t, J = 9.5Hz, 1H), 2.47 (s, 3H), 1.20 (t, J = 7.5Hz, 3H).

[0306] Example 22: Synthesis of Compound A77

[0307] Reference Example 18 Synthesis of Compound A79, using Substituting A79-1, compound A77 was synthesized. ESI m / z [MH] - =734.4. 1 H NMR(400MHz,DMSO-d6)δ13.33(s,1H),10.47(s,1H),8.61(s,1H),8.30(s,1H),8.18(s, 1H),8.09(d,J=8.6Hz,1H),8.00(d,J=2.1Hz,1H),7.92(s,2H),7.74(dd,J=8.7,2.1Hz,1 H),5.45(s,2H),4.57(d,J=12.5Hz,1H),3.56(t,J=10.8Hz,3H),3.29(d,J=12.9Hz,2H), 3.09–3.03(m,2H),2.89(d,J=11.2Hz,1H),2.78–2.66(m,1H),2.48(s,3H),1.25(s,3H).

[0308] Example 23: Synthesis of Compound A75

[0309] Reference Example 14 Synthesis of Compound A36, using Instead of dimethylamine, compound A75 was synthesized. ESI m / z [M+H] + =720.4. 1H NMR (400MHz, DMSO-d6) δ10.36(s,1H),8.59(s,1H),8.08(d,J=8.6Hz,1H),7.98(d, J=2.1Hz,1H),7.74(dd,J=8.8,2.1Hz,1H),5.23(s,2H),4.53(d,J=12.4Hz,1H),3.5 3(s,4H),3.27(d,J=11.7Hz,3H),2.99(s,6H),2.80(d,J=11.5Hz,1H),2.62(d,J=1 0.7Hz, 1H), 2.49 (d, J = 4.6Hz, 1H), 2.46 (s, 3H), 2.18 (s, 6H), 1.18 (t, J = 7.4Hz, 3H).

[0310] Example 24: Synthesis of Compound A20

[0311] 1) Synthesis of intermediate A20-2:

[0312] A20-1 (9.0 g, 44.53 mmol) and anhydrous tetrahydrofuran (225 mL) were added sequentially to a 1000 mL three-necked flask. Under nitrogen, the mixture was stirred at -70 to -75°C. A solution of n-butyllithium in n-hexane (2.5 mol / L, 48 mL, 120 mmol) was slowly added. The mixture was stirred at -70 to -75°C for 20 minutes. The temperature was slowly raised to -5-0°C, stirred at -5-0°C for 70 minutes, cooled to -70 to -75°C, and hexachloroethane (31.6 g, 133.6 mmol) was added at -70 to -75°C. The mixture was stirred at -70 to -75°C for 40 minutes, then slowly returned to room temperature and stirred for another 40 minutes. The reaction was quenched with 10% (mass fraction) aqueous citric acid solution and extracted twice with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to afford A20-2 (3.5 g, 33%).

[0313] 2) Synthesis of intermediate A20-3:

[0314] To a solution of A20-2 (3 g, 12.68 mmol) in anhydrous tert-butanol (30 mL) was added diphenylphosphoryl azide (4.08 g, 14.84 mmol). After the addition was complete, N,N-diisopropylethylamine (1.92 g, 19.02 mmol) was added to the reaction mixture. The reaction mixture was heated to 80°C and stirred overnight. The reaction mixture was concentrated under reduced pressure, and the residue was added to a saturated sodium bicarbonate solution. The mixture was extracted with ethyl acetate (3 × 50 mL). The organic phase was washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography to obtain compound A20-3 (2.8 g, 71%) as a white solid.

[0315] 3) Synthesis of intermediate A20-4:

[0316] A20-3 (450 mg, 1.46 mmol) was dissolved in dichloromethane (3 mL) and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at 25°C for 2 hours. LCMS showed that the reaction was complete. The reaction mixture was concentrated to give A20-4 (300 mg, mixture, yield: 98.8%) as a brown oil. ESI m / z [M+H] + =208.1.

[0317] 4) Synthesis of intermediate A20-5:

[0318] Compound A20-4 (0.3 g, 1.45 mmol) and potassium carbonate (0.6 g, 4.35 mmol) were dissolved in dichloromethane (2 mL). Bromoacetyl bromide (0.44 g, 2.17 mmol) was added dropwise with stirring at 0°C. The mixture was stirred at 25°C for 1 hour. LCMS indicated the reaction was complete. The reaction solution was poured into water, extracted with dichloromethane (50 mL x 2), and concentrated to afford A20-5 (0.3 g, 63%) as a yellow solid.

[0319] 5) Synthesis of intermediate A20-6:

[0320] To the reaction flask, A1-1 (5 g, 11.7 mmol), phenylboronic acid (2.85 g, 23.4 mmol), potassium phosphate (7.45 g, 35.1 mmol), methanesulfonic acid (2-dicyclohexylphosphino-2',4',6'-tri-isopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl) palladium (II) (0.5 g, 0.58 mmol), 1,4-dioxane (40 mL), and water (20 mL) were added in sequence. The atmosphere was purged with nitrogen three times, and the mixture was stirred at 100°C for 10 hours. LCMS showed that the reaction was complete, and the upper organic phase was concentrated to give an off-white solid crude product A20-6 (8.2 g, yield: 100%, theoretical content: 60%). m / z[M+H] + =425.2.

[0321] 6) Synthesis of intermediate A20-7:

[0322] Compound A20-6 (5 g, 5.9 mmol), A20-5 (3.1 g, 9.4 mmol), and N,N-diisopropylethylamine (2.3 g, 17.7 mmol) were dissolved in acetonitrile (80 mL) and stirred at 45°C for 10 hours. LCMS showed the reaction was complete. The reaction solution was filtered and washed with acetonitrile to give a white crude product A20-7 (4 g, yield: 75.4%). m / z [M-56] +=616.3.

[0323] 7) Synthesis of intermediate A20-8:

[0324] Compound A20-7 (4 g, 6.06 mmol) was dissolved in dichloromethane (30 mL) and trifluoroacetic acid (5 mL) was added dropwise. The reaction mixture was stirred at 25°C for 1 hour. LCMS showed that the reaction was complete. The reaction mixture was filtered and concentrated to give a yellow oil A20-8 (4 g, yield: 100%). m / z [M+H] + =572.3.

[0325] 8) Synthesis of product A20:

[0326] A1-7 (1 g, 6.49 mmol), HOBT (0.88 g, 6.43 mmol), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (1.54 g, 8.04 mmol) were dissolved in N,N-dimethylformamide (2 mL) and stirred at 25°C for 1 hour. Compound A20-8 (3 g, 5.36 mmol) and N,N-diisopropylethylamine (2.08 g, 16.08 mmol) were then added sequentially to the reaction solution, which was stirred at 25°C for another 1 hour. LCMS indicated the reaction was complete. The reaction solution was added to water, extracted with dichloromethane, washed with water and saturated brine, dried over sodium sulfate, filtered, concentrated, and purified on a silica gel column (dichloromethane / ethyl acetate) to yield the white product A20 (1.45 g, yield: 38.88%, purity: 97.82%). m / z [M+H] + =708.4. 1 HNMR(400MHz,DMSO-d6)δ10.39(s,1H),8.60(s,1H),8.15(dd,J=6.8,3.0Hz ,2H),7.56(dd,J=5.1,2.0Hz,3H),7.44(s,2H),5.32(s,2H),4.56(d,J=12. 6Hz,1H),3.55(d,J=12.0Hz,3H),3.27(s,1H),3.04(dd,J=13.5,9.0Hz,3H) ,2.90–2.81(m,1H),2.69(d,J=10.7Hz,1H),2.47(s,3H),1.32–1.24(m,3H).

[0327] Example 25: Synthesis of Compound A3

[0328] Referring to the synthesis of compound A26 in Example 15, 2-methyl-2H-indazole-6-boronic acid was used instead of A26-1 to synthesize compound A3. m / z [M+H] + =750.4. 1 H NMR (400MHz, DMSO-d6) δ10.48(s,1H),8.61(s,1H),8.44(s,1H),8.37(d,J=1.2Hz,1H),8.08( d,J=8.6Hz,1H),8.00(d,J=2.1Hz,1H),7.89–7.80(m,2H),7.74(dd,J=8.8,2.1Hz,1H),5.45(s ,2H),4.57(d,J=12.4Hz,1H),4.23(s,3H),3.57(td,J=12.4,11.8,6.6Hz,4H),3.05(dd,J=15 .8,9.5Hz,3H),2.88(d,J=11.3Hz,1H),2.76–2.67(m,1H),2.48(s,3H),1.26(d,J=7.3Hz,3H).

[0329] Example 26: Synthesis of Compound A7

[0330] Referring to the synthesis of compound A26 in Example 15, 1-methyl-1H-indazole-6-boronic acid was used instead of A26-1 to synthesize compound A7. m / z [MH] - =748.5. 1 H NMR (400MHz, DMSO-d6) δ10.50(s,1H),8.62(s,1H),8.35(s,1H),8.15(s,1H),8.11(d,J=8.6 Hz,1H),8.01(d,J=2.0Hz,1H),7.98–7.90(m,2H),7.76(dd,J=8.8,2.1Hz,1H),5.48(s,2H),4 .58(d,J=12.5Hz,1H),4.15(s,3H),3.58(q,J=11.7,10.8Hz,3H),3.28(s,1H),3.07(d,J=9. 0Hz, 3H), 2.90 (d, J = 11.3Hz, 1H), 2.72 (d, J = 11.0Hz, 1H), 2.49 (s, 3H), 1.25 (d, J = 7.6Hz, 3H).

[0331] Example 27: Synthesis of Compound A8

[0332] Referring to the synthesis of compound A26 in Example 15, (2,2-difluorobenzo[d][1,3]dioxol-5-yl)boronic acid was used instead of A26-1 to synthesize compound A8. m / z [MH] - =774.4. 1 H NMR (400MHz, DMSO-d6) δ10.45(s,1H),8.60(s,1H),8.08(d,J=8.6Hz,1H),8.04(d,J=1.5Hz ,1H),8.03–7.99(m,2H),7.74(dd,J=8.8,2.1Hz,1H),7.60(d,J=8.4Hz,1H),5.42(s,2H),4 .56(d,J=12.3Hz,1H),3.56(dd,J=14.1,10.7Hz,3H),3.32–3.25(m,2H),3.03(dd,J=14.8, 8.4Hz,3H),2.87(d,J=11.1Hz,1H),2.72–2.66(m,1H),2.47(s,3H),1.24(t,J=7.4Hz,3H).

[0333] Example 28: Synthesis of Compound A21

[0334] Referring to the synthesis of compound A1 in Example 1, compound A21 was synthesized by replacing A1-4 with A20-5. m / z[M+H] + =806.4. 1 H NMR (400MHz, DMSO-d6) δ10.37(s,1H),8.55(s,1H),7.94(d,J=8.6Hz,2H),7.41( s,2H),7.04(d,J=8.6Hz,2H),5.27(s,2H),4.53(d,J=12.3Hz,1H),3.53(d,J=3.1 Hz,2H),3.49(d,J=11.2Hz,4H),3.27–3.25(m,4H),3.01(d,J=8.8Hz,4H),2.82(d ,J=11.1Hz,2H),2.71–2.61(m,2H),2.44(s,3H),2.23(s,3H),1.23–1.19(m,3H).

[0335] Example 29: Synthesis of Compound A30

[0336] 1) Synthesis of intermediate A30-2:

[0337] Compound A30-1 (2 g, 12.89 mmol) and NCS (2.53 g, 18.95 mmol) were dissolved in tetrahydrofuran (20 mL), replaced with nitrogen three times, and cooled to -76°C. LiHMDS (1 M, 19.34 mL) was added dropwise under temperature control and stirred for two hours. LCMS showed that the reaction was complete. The reaction solution was washed with water and purified on a silica gel column to obtain the white product A30-2 (1.2 g, 49%). m / z [M+H] + =190.1.

[0338] 2) Synthesis of intermediate A30-3:

[0339] Compound A30-2 (200 mg, 1.05 mmol) and ammonium chloride (280 mg, 5.25 mmol) were dissolved in a mixture of ethyl acetate (2 mL) and water (1 mL), followed by the addition of zinc powder (210 mg, 3.15 mmol). The mixture was stirred at 25°C for 4 hours. LCMS indicated the reaction was complete, and the reaction solution was concentrated and purified on a silica gel column to yield the white product A30-3 (120 mg, 71.2%). m / z [M+H] + =160.2.

[0340] 3) Synthesis of product A30:

[0341] Referring to the synthesis of compound A9 in Example 7, compound A30 was synthesized by replacing A9-1 with A30-3. m / z[M+H] + =666.4. 1 H NMR(400MHz,DMSO-d6)δ10.18(s,1H),8.58(s,1H),7.78(s,1H),6.92–6.80(m,1H),5.19 (s,2H),4.64(p,J=6.6Hz,1H),4.54(d,J=12.5Hz,1H),4.28(p,J=2.9Hz,2H),3.83(t,J=5.3Hz,2H),3.60(s,2H),3.27(d,J=2.7Hz,4H) ,2.99(d,J=12.4Hz,3H),2.84(d,J=10.4Hz,1H),2.66(d,J=10.9Hz,1H),2.46(s,3H),1.40(s,3H),1.38(s,3H),1.20(t,J=7.5Hz,3H).

[0342] Example 30: Synthesis of Compound A32

[0343] Referring to the synthesis of compound A31 in Example 3, compound A32 was synthesized by replacing A31-1 with A32-1. m / z [MH] - =702.4. 1 H NMR (400MHz, DMSO-d6) δ10.39(s,1H),8.99(s,1H),8.09(d,J=8.3Hz,2H),7.99(s,1H),6.86(s,1H),5.35(s,2H),4.28(s, 3H),3.82(t,J=5.1Hz,3H),3.74(s,3H),3.48(d,J=11.1Hz,4H),3.08–2.98(m,4H),2.79(s,2H),2.17(s,3H),1.23(s,3H).

[0344] Example 31: Synthesis of Compound A47

[0345] Referring to the synthesis of compound A36 in Example 14, compound A47 was synthesized by using 2-oxa-6-azaspiro[3.3]heptane instead of dimethylamine. m / z[M+H] + =717.3. 1 H NMR (400MHz, DMSO-d6) δ10.38(s,1H),8.54(s,1H),8.09(d,J=8.6Hz,1H),7.99(d,J= 2.1Hz,1H),7.75(dd,J=8.8,2.2Hz,1H),5.24(s,2H),4.72(s,3H),4.53(d,J=12.4Hz ,1H),4.16(s,3H),3.52(s,6H),2.97(q,J=10.4,7.8Hz,3H),2.79(d,J=11.5Hz,1H), 2.72–2.57(m,1H),2.45(s,2H),2.02(dt,J=13.6,7.0Hz,1H),1.17(t,J=7.4Hz,3H).

[0346] Example 32: Synthesis of Compound A70

[0347] 1) Synthesis of intermediate A70-1:

[0348] A71-4 (120 mg, 0.44 mmol) was dissolved in N,N-dimethylformamide (2 mL), and N-chlorosuccinimide (58.7 mg, 0.44 mmol) was added. The mixture was stirred at room temperature for 2 hours. After LCMS showed that the reaction was complete, the reaction solution was diluted with water (10 mL) and extracted with ethyl acetate (20 mL × 3). The combined organic phases were washed with saturated brine (10 mL × 2), dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1:1 (volume ratio)) to obtain A70-1 (180 mg) as a light yellow solid. m / z [M+H] + =307.0,309.0.

[0349] 2) Synthesis of product A70:

[0350] Referring to the synthesis of compound A1 in Example 1, compound A70 was synthesized by replacing A1-2 with phenylboronic acid and A70-1 with A1-4. m / z [M+H] + =687.2. 1 H NMR (400MHz, DMSO-d6) δ8.46(s,1H),8.08(d,J=5.9Hz,3H),7.70(d,J=7.7Hz,1H),7.57-7.46(m,4H),7.04(d,J=7.7Hz,1H),5.42(s,2H),5.08–4. 97(m,1H),3.54-3.48(m,2H),3.04-2.94(m,6H),2.86-2.80(m,1H),2.69 -2.62(m,1H),2.40(s,3H),1.26(d,J=6.7Hz,6H),1.17(t,J=7.2Hz,3H).

[0351] Example 33: Synthesis of Compound A71

[0352] 1) Synthesis of intermediate A71-2:

[0353] A71-1 (950 mg, 6.78 mmol) and potassium carbonate (1.88 g, 13.4 mmol) were dissolved in acetonitrile (10 mL). 2-Iodopropane (5.77 g, 33.9 mmol) was added under ice-cooling and stirred at this temperature for 1 hour. After LCMS indicated the reaction was complete, the mixture was slowly poured into water (10 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the solvent was removed by evaporation under reduced pressure. The residue was purified by silica gel column chromatography to afford A71-2 (910 mg, 73.0% yield) as a yellow oil. m / z [M+H] + =183.1.1 HNMR (400MHz, DMSO-d6) δ (ppm) 8.09 (d, J = 7.6Hz, 1H), 7.11 (d, J = 2.7Hz, 1H), 6.88 (dd, J = 7.6, 2.7Hz, 1H), 5.03 (p, J = 6.8Hz, 1H), 1.33 (d, J = 6.8Hz, 6H).

[0354] 2) Synthesis of intermediate A71-3:

[0355] A71-2 (450 mg, 2.50 mmol) was dissolved in ethyl acetate (10 mL), wet palladium on carbon (50 mg, 10%) was added, and the mixture was stirred at room temperature under a hydrogen balloon for 12 hours. After LCMS showed that the reaction was complete, the reaction solution was filtered through celite, and the filtrate was evaporated under reduced pressure to remove the solvent to give crude A71-3 (370 mg) as a white solid. m / z [M+H] + =153.1.

[0356] 3) Synthesis of intermediate A71-4:

[0357] A71-3 (350 mg, 2.30 mmol) and N,N-diisopropylethylamine (590 mg, 4.60 mmol) were dissolved in dichloromethane (5 mL). Bromoacetyl bromide (520 mg, 2.60 mmol) was added under ice-cooling and stirred for 1 hour. After LCMS showed that the reaction was complete, the mixture was slowly poured into water (10 mL) and extracted with dichloromethane (20 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the solvent was removed by evaporation under reduced pressure. The residue was purified by silica gel column chromatography to obtain A71-4 (210 mg, 33.43% yield) as a yellow oil. m / z [M+H] + =273.1 275.1.

[0358] 4) Synthesis of product A71:

[0359] Referring to the synthesis of compound A1 in Example 1, phenylboronic acid was used instead of A1-2, and A71-4 was used instead of A1-4 to synthesize compound A71. m / z [M+H] + =687.2. 1H NMR(400MHz,DMSO-d6)δ11.80(s,1H),9.52(s,1H),8.59(s,1H),8.22(s,2 H),7.47(s,3H),6.73(d,J=7.7Hz,1H),6.60(s,1H),5.73-5.61(m,1H),5. 25-5.05(m,3H),3.83(t,J=11.6Hz,2H),3.55-3.42(m,1H),3.19–2.74(m, 4H),2.57(s,3H),2.34-2.17(m,1H),2.04-1.96(m,1H),1.33-1.28(m,9H).

[0360] Example 34: Synthesis of Compound A81

[0361] Referring to the synthesis of compound A1 in Example 1, compound A81 was synthesized by replacing A1-2 with 1-methyl-1H-indazole-5-boronic acid and A20-5 with A1-4. m / z [MH] - =760.4. 1 H NMR(400MHz,DMSO-d6)δ10.42(s,1H),8.67–8.54(m,2H),8.24–8.13(m,2H),7 .81(d,J=8.9Hz,1H),7.45(s,2H),5.34(s,2H),4.57(d,J=12.4Hz,1H),4.12(s ,3H),3.56(d,J=11.2Hz,3H),3.30–3.24(m,1H),3.05(d,J=10.1Hz,3H),2.87 (d,J=11.1Hz,1H),2.69(d,J=10.3Hz,1H),2.47(s,3H),1.27(d,J=7.1Hz,3H).

[0362] Example 35: Synthesis of Compound A82

[0363] Referring to the synthesis of compound A1 in Example 1, 2-methyl-2H-indazole-5-boronic acid was used instead of A1-2, and A20-5 was used instead of A1-4 to synthesize compound A82. m / z [MH] - =760.4. 1H NMR (400MHz, DMSO-d6) δ10.41(s,1H),10.27(s,1H),8.61(s,1H),8.58–8.52(m,2H),8. 01(dd,J=9.1,1.6Hz,1H),7.74(d,J=9.1Hz,1H),7.45(s,2H),5.33(s,2H),4.57(d,J=1 2.5Hz,1H),4.23(s,3H),3.64–3.47(m,3H),3.28(d,J=12.7Hz,1H),3.05(d,J=9.3Hz,3 H), 2.87 (d, J = 11.0Hz, 1H), 2.69 (d, J = 10.4Hz, 1H), 2.48 (s, 3H), 1.25 (d, J = 7.9Hz, 3H).

[0364] Example 36: Synthesis of Compound A91

[0365] Referring to the synthesis of compound A26 in Example 15, quinoline-6-pinacol boronate was used instead of A26-1 to synthesize compound A91. m / z [MH] - =745.4. 1 H NMR (400MHz, DMSO-d6) δ10.48(s,1H),9.00(dd,J=4.2,1.7Hz,1H),8.79(d,J=1.9Hz,1H),8.62(s,1H),8.58(dd,J=8.4 ,1.7Hz,1H),8.49(dd,J=8.8,1.9Hz,1H),8.18(d,J=8.8Hz,1H),8.12(d,J=8.6Hz,1H),8.01(d,J=2.1Hz,1H),7.74(dd ,J=8.9,2.1Hz,1H),7.64(dd,J=8.3,4.2Hz,1H),5.48(s,2H),4.58(d,J=12.5Hz,1H),3.58(td,J=12.6,11.7,7.0Hz,3 H),3.28(s,2H),3.06(p,J=9.3,8.4Hz,3H),2.90(d,J=11.3Hz,1H),2.80–2.67(m,1H),2.48(s,3H),1.28–1.24(m,3H).

[0366] Example 37: Synthesis of Compound A92

[0367] Referring to the synthesis of compound A26 in Example 15, compound A92 was synthesized by using benzo[d]thiazole-6-boronic acid instead of A26-1. m / z [MH]- =751.4. 1 H NMR (400MHz, DMSO-d6) δ10.47(s,1H),9.52(s,1H),8.94(d,J=1.6Hz,1H),8.61(s,1H),8.31(dd, J=8.6,1.7Hz,1H),8.23(d,J=8.5Hz,1H),8.10(d,J=8.6Hz,1H),8.00(d,J=2.0Hz,1H),7.74(dd, J=8.8,2.1Hz,1H),5.45(s,2H),4.57(d,J=12.4Hz,1H),3.65–3.51(m,3H),3.30(s,2H),3.06(d, J=9.5Hz,3H),2.89(d,J=10.9Hz,1H),2.71(d,J=10.8Hz,1H),2.48(s,3H),1.25(t,J=7.4Hz,3H).

[0368] Example 38: Synthesis of Compound A136

[0369] Referring to the synthesis of compound A26 in Example 15, pyrazolo[1,5-a]pyridine-5-pinacol boronate was used instead of A26-1 to synthesize compound A136. m / z [MH] - =734.4. 1 H NMR(400MHz,DMSO-d6)δ10.39(s,2H),8.77(d,J=7.3Hz,1H),8.56(s,1H),8.40(s,1H),8.15– 8.01(m,2H),7.94(d,J=2.1Hz,1H),7.69(dd,J=9.1,2.0Hz,1H),7.49(dd,J=7.3,1.8Hz,1H),6 .80(d,J=2.2Hz,1H),5.39(s,2H),4.52(d,J=12.6Hz,1H),3.52(d,J=11.4Hz,3H),3.01(d,J=8 .8Hz,3H),2.84(d,J=11.3Hz,1H),2.66(d,J=10.8Hz,1H),2.43(s,3H),1.20(d,J=7.8Hz,3H).

[0370] Example 39: Synthesis of Compound A139

[0371] Referring to the synthesis of compound A26 in Example 15, benzo[b]thiophene-2-boronic acid was used instead of A26-1 to synthesize compound A139. m / z [MH]- =750.3. 1 H NMR (400MHz, DMSO-d6) δ10.45(s,1H),8.61(s,1H),8.16(s,1H),8.09(dd,J=11.5,7.5Hz,2 H),8.00(q,J=4.3,3.5Hz,2H),7.74(dd,J=8.8,2.1Hz,1H),7.52–7.43(m,2H),5.42(s,2H), 4.57(d,J=12.4Hz,1H),3.55(dd,J=11.8,3.3Hz,3H),3.29(d,J=17.3Hz,2H),3.05(d,J=9.9 Hz,3H),2.88(d,J=11.2Hz,1H),2.70(d,J=10.2Hz,1H),2.47(s,3H),1.24(t,J=7.3Hz,3H).

[0372] Example 40: Synthesis of Compound A141

[0373] 1) Synthesis of intermediate A141-4:

[0374] Referring to the synthesis of compound A20-5 in Example 24, NFSI was used instead of hexachloroethane in the first step to synthesize compound A141-4.

[0375] 2) Synthesis of intermediate A141-5:

[0376] Compound A1-1 (100 mg, 0.24 mmol), A141-4 (82 mg, 0.26 mmol), and N,N-diisopropylethylamine (93 mg, 0.72 mmol) were dissolved in acetonitrile (2 mL) and stirred at 45°C for 8 hours. LCMS showed the reaction was complete. The reaction solution was poured into water, extracted with dichloromethane, and concentrated to afford A141-5 (130 mg, yield: 87%) as a yellow solid. m / z [M+H] + =658.1.

[0377] 2) Synthesis of product A141:

[0378] Referring to the synthesis of compound A26 in Example 15, compound A141 was synthesized by replacing A36-1 with A141-5 and A26-1 with phenylboronic acid. m / z [M+H] + =692.4. 1H NMR (400MHz, DMSO-d6) δ10.56(s,1H),8.57(s,1H),8.13(dd,J=6.6,3.0Hz, 2H),7.60–7.48(m,4H),7.29(d,J=8.8Hz,1H),5.32(s,2H),4.56(d,J=12.6H z,1H),3.54(d,J=10.5Hz,3H),3.03(q,J=8.3Hz,3H),2.86(d,J=11.3Hz,1H ),2.71–2.66(m,1H),2.46(s,3H),2.10-1.96(m,1H),1.23(t,J=7.4Hz,3H).

[0379] Example 41: Synthesis of Compound A142

[0380] 1) Synthesis of intermediate A142-2:

[0381] Compound A142-1 (0.6 g, 2.38 mmol) and methylboronic acid (1.42 g, 23.8 mmol) were dissolved in a mixed solution of 1,4-dioxane (5 mL) and water (0.5 mL). Potassium carbonate (0.99 g, 7.14 mmol) was added with stirring. After three gas replacements, 1,1-bis(diphenylphosphino)ferrocenepalladium(II) chloride (0.17 g, 0.24 mmol) was added and stirred at 80°C for 16 hours. LCMS showed that the reaction was complete. The reaction solution was concentrated and purified by column chromatography (ethyl acetate / petroleum ether) to obtain A142-2 as a white solid (0.4 g, yield: 89.8%). m / z [M+H] + =188.1.

[0382] 2) Synthesis of intermediate A142-3:

[0383] Refer to the synthesis of compound A9-2 in Example 7, and replace A9-1 with A142-2 to obtain A142-3.

[0384] 3) Synthesis of product A142:

[0385] Referring to the synthesis of compound A1 in Example 1, compound A142 was synthesized by replacing A1-2 with phenylboronic acid and replacing A1-4 with A142-3. m / z [M+H] + =688.4. 1H NMR (400MHz, DMSO-d6) δ10.14(s,1H),8.61(s,1H),8.19–8.10(m,2H),7.57(dd,J=5. 2,2.0Hz,3H),7.24(d,J=8.7Hz,1H),7.16(d,J=8.7Hz,1H),5.28(s,2H),4.57(d,J=12 .5Hz,1H),3.60–3.53(m,3H),3.30(s,1H),3.06(td,J=13.5,12.0,8.4Hz,3H),2.87( d,J=10.7Hz,1H),2.73–2.66(m,1H),2.48(s,3H),2.22(s,3H),1.26(d,J=3.4Hz,3H).

[0386] Example 42: Synthesis of Compound A143

[0387] 1) Synthesis of intermediate A143-1:

[0388] Refer to the synthesis of compound A9-2 in Example 7, and use A142-1 instead of A9-1 to obtain A143-1.

[0389] 3) Synthesis of product A143:

[0390] Referring to the synthesis of compound A1 in Example 1, compound A143 was synthesized by replacing A1-2 with phenylboronic acid and A143-1 with A1-4. m / z [M+H] + =752.1,754.1. 1 H NMR(400MHz,DMSO-d6)δ10.35(s,1H),8.59(s,1H),8.16(dd,J=6.9,2.9Hz,2H),7 .56(dd,J=5.2,1.9Hz,3H),7.46(d,J=8.7Hz,1H),7.37(d,J=8.7Hz,1H),5.31(s,2 H),4.56(d,J=12.6Hz,1H),3.54(d,J=13.8Hz,3H),3.03(dd,J=15.4,9.4Hz,3H),2 .87(d,J=11.3Hz,1H),2.69(d,J=10.6Hz,1H),2.47(s,3H),1.27(d,J=7.0Hz,3H).

[0391] Example 43: Synthesis of Compound A144

[0392] 1) Synthesis of intermediate A144-1:

[0393] Compound A1-1 (100 mg, 0.23 mmol), A9-2 (76 mg, 0.26 mmol), and N,N-diisopropylethylamine (93 mg, 0.72 mmol) were dissolved in acetonitrile (2 mL) and stirred at 45°C for 8 hours. LCMS showed the reaction was complete. The reaction solution was poured into water, extracted with dichloromethane, and concentrated to obtain A144-1 (125 mg, yield: 85%) as a yellow solid. m / z [M+H] + =640.1,642.1.

[0394] 2) Synthesis of product 144:

[0395] Referring to the synthesis of compound A26 in Example 15, compound A144 was synthesized by replacing A36-1 with A144-1 and A26-1 with phenylboronic acid. m / z [M+H] + =674.4. 1 H NMR(400MHz,DMSO-d6)δ10.83(s,1H),8.59(s,1H),8.11(dd,J=6.6,3.0Hz,2H), 7.75(d,J=2.0Hz,1H),7.61–7.52(m,3H),7.41(d,J=8.8Hz,1H),7.30(dd,J=8.8, 2.1Hz,1H),5.26(s,2H),4.56(d,J=12.6Hz,1H),3.66–3.55(m,4H),3.06–2.97( m,3H),2.87(d,J=11.3Hz,1H),2.70(d,J=10.4Hz,1H),2.47(s,3H),1.22(s,3H).

[0396] Example 44: Synthesis of Compound A145

[0397] 1) Synthesis of intermediate A145-1:

[0398] Compound A1-1 (100 mg, 0.23 mmol), A20-5 (85 mg, 0.26 mmol), and N,N-diisopropylethylamine (93 mg, 0.72 mmol) were dissolved in acetonitrile (2 mL) and stirred at 45°C for 8 hours. LCMS showed the reaction was complete. The reaction solution was poured into water, extracted with dichloromethane, and concentrated to obtain A145-1 (132 mg, yield: 85%) as a yellow solid. m / z [M+H] + =674.1,676.1.

[0399] 2) Synthesis of product 145:

[0400] Referring to the synthesis of compound A26 in Example 15, compound A145 was synthesized by replacing A36-1 with A145-1 and replacing A26-1 with thiophene-2-pinacol boronate. m / z [M+H] + =714.3. 1 H NMR(400MHz,DMSO-d6)δ10.39(s,1H),8.59(s,1H),7.84–7.76(m,2H),7.44 (s,2H),7.24(dd,J=5.0,3.6Hz,1H),5.28(s,2H),4.56(d,J=12.5Hz,1H),3. 58–3.50(m,3H),3.29(d,J=2.9Hz,1H),3.04(q,J=11.8,9.1Hz,3H),2.86(d ,J=11.2Hz,1H),2.68(d,J=11.2Hz,1H),2.47(s,3H),1.24(t,J=7.5Hz,3H).

[0401] Example 45: Synthesis of Compound A146

[0402] Referring to the synthesis of compound A26 in Example 15, compound A146 was synthesized by replacing A36-1 with A145-1 and replacing A26-1 with furan-2-pinacol boronate. m / z [MH] - =696.4. 1 H NMR(400MHz,DMSO-d6)δ10.38(s,1H),8.60(s,1H),7.94(s,1H),7.44(s,2H ),7.17(d,J=3.4Hz,1H),6.76–6.69(m,1H),5.29(s,2H),4.56(d,J=12.5Hz ,1H),3.53(d,J=10.6Hz,3H),3.32–3.27(m,1H),3.03(q,J=8.1Hz,3H),2.8 7–2.83(m,1H),2.68(d,J=11.0Hz,1H),2.47(s,3H),1.24(d,J=6.7Hz,3H).

[0403] Example 46: Synthesis of Compound A147

[0404] Referring to the synthesis of compound A26 in Example 15, compound A147 was synthesized by replacing A36-1 with A145-1 and A26-1 with 4-dimethylaminophenylboronic acid. ESI (m / z) = 751.4 [M+H] + . 1H NMR (400MHz, chloroform-d) δ11.80(s,1H),9.17(s,1H),8.59(s,1H),8.17–8.06(m,2H),8.00( d,J=8.9Hz,1H),6.96(d,J=8.9Hz,1H),6.73(d,J=8.9Hz,2H),5.66(d,J=13.0Hz,1H), 5.12(s,2H),4.79(d,J=12.8Hz,1H),3.83(dd,J=13.1,10.2Hz,2H),3.50(s,1H),3.21 (q,J=7.6Hz,2H),3.04(s,7H),2.90–2.71(m,2H),2.57(s,3H),1.34(t,J=7.5Hz,3H).

[0405] Example 47: Synthesis of Compound A148

[0406] Referring to the synthesis of compound A26 in Example 15, compound A148 was synthesized by replacing A36-1 with A145-1 and replacing A26-1 with 4-(1-pyrrolidinyl)-phenylboronic acid pinacol ester. m / z [M+H] + =777.5. 1 H NMR (400MHz, chloroform-d) δ11.79(s,1H),9.21(s,1H),8.58(s,1H),8.14–8.08(m,2H),8.00(d,J=8.9Hz ,1H),6.95(d,J=8.9Hz,1H),6.62–6.55(m,2H),5.65(d,J=13.1Hz,1H),5.11(s,2H),4.79(d,J=12 .7Hz,1H),3.88–3.77(m,2H),3.51(d,J=12.6Hz,1H),3.42–3.31(m,4H),3.21(q,J=7.5Hz,2H),3 .07(s,1H),2.79(dd,J=33.3,11.7Hz,2H),2.56(s,3H),2.06–2.01(m,4H),1.34(t,J=7.5Hz,3H).

[0407] Example 48: Synthesis of Compound A149

[0408] Referring to the synthesis of compound A26 in Example 15, compound A149 was synthesized by replacing A36-1 with A145-1 and A26-1 with 4-methoxyphenylboronic acid. m / z [MH] - =736.5. 1H NMR (400MHz, DMSO-d6) δ10.40(s,1H),8.60(s,1H),8.10–8.06(m,2H),7.44(s,2H),7.15–7.08(m,2H),5.30(s,2H),4.56(d,J=12.4Hz,1H ),3.86(s,3H),3.53(s,3H),3.27(s,1H),3.04(d,J=9.2Hz,3H),2.86(d,J=10.6Hz,1H),2.68(d,J=11.5Hz,1H),2.47(s,3H),1.24(s,3H).

[0409] Example 50: Synthesis of Compound A151

[0410] Referring to the synthesis of compound A26 in Example 15, 1-methyl-1H-indole-5-boronic acid pinacol ester was used instead of A26-1 to synthesize compound A151. m / z [M+H] + =749.4. 1 H NMR (400MHz, DMSO-d6) δ10.44(s,1H),8.58(s,1H),8.38–8.33(m,1H),8.07(d,J=8.5Hz,1H),8.01– 7.90(m,2H),7.71(d,J=8.6Hz,1H),7.55(d,J=8.7Hz,1H),7.40(d,J=3.1Hz,1H),6.54(d,J=3.1Hz, 1H),5.41(s,2H),4.54(d,J=12.4Hz,1H),3.83(s,3H),3.55(q,J=11.3Hz,3H),3.25(s,1H),3.02(d ,J=11.0Hz,3H),2.85(d,J=11.3Hz,1H),2.67(d,J=9.4Hz,1H),2.45(s,3H),1.22(t,J=7.3Hz,3H).

[0411] Example 51: Synthesis of Compound A152

[0412] Referring to the synthesis of compound A26 in Example 15, 1-methyl-1H-benzo[d]imidazol-5-boronic acid pinacol ester was used instead of A26-1 to synthesize compound A152. m / z [M+H] + =750.4. 1H NMR(400MHz,DMSO-d6)δ10.95(s,1H),9.06(s,1H),8.86(s,1H),8.76(s,1H),8.5 5(t,J=8.9Hz,2H),8.45(s,1H),8.18(d,J=8.6Hz,2H),5.90(s,2H),5.02(d,J=12 .2Hz,1H),4.36(s,3H),4.01(s,3H),3.75(s,1H),3.50(p,J=10.6,9.1Hz,3H),3. 33(d,J=11.3Hz,1H),3.15(d,J=10.7Hz,1H),2.93(s,3H),1.70(t,J=7.4Hz,3H).

[0413] Example 52: Synthesis of Compound A153

[0414] Referring to the synthesis of compound A26 in Example 15, compound A153 was synthesized by using benzo[d]thiazole-5-boronic acid pinacol ester instead of A26-1. m / z [MH] - =751.4. 1 H NMR (400MHz, DMSO-d6) δ10.48(s,1H),9.50(s,1H),8.76(d,J=1.5Hz,1H),8.58(s,1H),8.34 (d,J=8.4Hz,1H),8.23(dd,J=8.4,1.6Hz,1H),8.06(d,J=8.6Hz,1H),7.98(d,J=2.0Hz,1H),7 .72(dd,J=8.8,2.1Hz,1H),5.44(s,2H),4.55(d,J=12.6Hz,1H),3.53(t,J=11.4Hz,4H),3.11 –3.00(m,3H),2.86(d,J=11.3Hz,1H),2.74–2.65(m,1H),2.45(s,3H),1.24(d,J=6.9Hz,3H).

[0415] Example 53: Synthesis of Compound A154

[0416] Referring to the synthesis of compound A26 in Example 15, compound A154 was synthesized by replacing A36-1 with A145-1 and quinoline-6-pinacol boronate with A26-1. m / z [M+H] + =759.4. 1H NMR (400MHz, chloroform-d) δ11.81(s,1H),8.98(dd,J=4.3,1.6Hz,1H),8.87(s,1H),8.80(d,J=1.8Hz,1H),8.59(d,J =6.5Hz,2H),8.29–8.23(m,1H),8.20(d,J=8.8Hz,1H),8.03(d,J=9.0Hz,1H),7.47(dd,J=8.3,4.3Hz,1H),6.9 8(d,J=9.0Hz,1H),5.70(d,J=13.0Hz,1H),5.20(s,2H),4.82(d,J=13.0Hz,1H),3.86(t,J=11.7Hz,2H),3.52( s,1H),3.24(d,J=7.9Hz,2H),3.09(s,1H),2.84(dd,J=32.9,11.9Hz,2H),2.57(s,3H),1.38(t,J=7.5Hz,3H).

[0417] Example 54: Synthesis of Compound A155

[0418] Referring to the synthesis of compound A26 in Example 15, compound A155 was synthesized by replacing A36-1 with A145-1 and replacing A26-1 with benzo[d]thiazole-6-boronic acid pinacol ester. m / z[M+H] + =765.4. 1 H NMR (400MHz, chloroform-d) δ11.80(s,1H),9.10(s,1H),8.93(d,J=1.6Hz,1H),8.85–8.77(m,1H),8.60(s, 1H),8.44(dd,J=8.6,1.7Hz,1H),8.22(d,J=8.6Hz,1H),8.02(d,J=8.9Hz,1H),6.98(d,J=8.9Hz,1 H),5.70(d,J=12.8Hz,1H),5.18(s,2H),4.82(d,J=12.6Hz,1H),3.85(t,J=11.7Hz,2H),3.51(s,1 H), 3.23 (d, J = 7.8Hz, 2H), 3.09 (s, 1H), 2.84 (d, J = 26.3Hz, 2H), 2.57 (s, 3H), 1.37 (t, J = 7.5Hz, 3H).

[0419] Example 55: Synthesis of Compound A156

[0420] Referring to the synthesis of compound A26 in Example 15, compound A156 was synthesized by replacing A36-1 with A145-1 and replacing A26-1 with 1-methylpyridin-6(1H)-one-3-boronic acid pinacol ester. m / z [M+H] + =739.4. 1 H NMR (400MHz, chloroform-d) δ11.78(s,1H),8.79(s,1H),8.59(s,1H),8.35(d,J=2.4Hz,1H),8.12(dd,J=9.5, 2.5Hz,1H),7.94(d,J=8.9Hz,1H),6.99(d,J=8.9Hz,1H),6.69(d,J=9.5Hz,1H),5.68(d,J=13.1Hz,1H ),5.16(s,2H),4.80(d,J=12.8Hz,1H),3.82(t,J=11.6Hz,2H),3.63(s,3H),3.47(d,J=19.3Hz,1H),3 .17(d,J=7.7Hz,2H),3.07(s,1H),2.82(dd,J=32.4,11.7Hz,2H),2.57(s,3H),1.34(t,J=7.5Hz,3H).

[0421] Example 56: Synthesis of Compound A157

[0422] Referring to the synthesis of compound A26 in Example 15, compound A157 was synthesized by substituting A145-1 for A36-1 and 4-(4-(oxetan-3-yl)piperazin-1-yl)phenylboronic acid pinacol ester for A26-1. ESI (m / z) = 848.5 [M+H] + . 1 H NMR (400MHz, chloroform-d) δ9.07(s,1H),8.58(s,1H),8.14(d,J=8.8Hz,2H),7.99(d,J=8.9Hz,1H),6.95(t ,J=8.6Hz,3H),5.66(d,J=12.9Hz,1H),5.13(s,2H),4.79(d,J=12.8Hz,1H),4.69(dt,J=16.7,6.3H z,4H),3.82(t,J=11.4Hz,2H),3.61–3.42(m,2H),3.36(t,J=4.9Hz,4H),3.21(q,J=7.5Hz,2H),3.0 7(s,1H),2.80(dd,J=32.8,11.5Hz,2H),2.56(s,3H),2.50(t,J=5.0Hz,4H),1.34(t,J=7.5Hz,3H).

[0423] Example 57: Synthesis of Compound A158

[0424] Referring to the synthesis of compound A26 in Example 15, compound A158 was synthesized by replacing A36-1 with A145-1 and A26-1 with 6-(4-methylpiperazin-1-yl)pyridine-3-boronic acid pinacol ester. m / z [M+H] + =807.4. 1 H NMR(400MHz,DMSO-d6)δ10.36(s,1H),8.79(d,J=2.4Hz,1H),8.57(s,1H),8.19–8.08(m,1H ),7.40(s,2H),6.96(d,J=9.1Hz,1H),5.26(s,2H),4.53(d,J=12.3Hz,1H),3.60(t,J=5.1H z,4H),3.52(s,2H),3.26–3.23(m,2H),2.99(t,J=8.6Hz,3H),2.82(d,J=11.1Hz,1H),2.64 (d,J=11.0Hz,1H),2.44(s,3H),2.41(t,J=5.0Hz,4H),2.23(s,3H),1.21(t,J=7.5Hz,3H).

[0425] Example 58: Synthesis of Compound A160

[0426] 1) Synthesis of intermediate A160-2:

[0427] To a solution of A160-1 (1 g, 4.11 mol) in acetonitrile (10 mL) was added diisopropylethylamine (170 mg, 12.33 mmol) and A20-5 (1.49 g, 4.52 mmol), and the mixture was stirred at room temperature overnight. The reaction mixture was dried and separated by column chromatography to afford A160-2 (1.6 g, 80%).

[0428] 2) Synthesis of intermediate A160-3:

[0429] A160-2 (0.5 g, 1.02 mmol), 6-quinolinylboronic acid pinacol ester (0.31 g, 1.22 mmol), Pd(dppf)Cl2 (75 mg, 0.1 mmol), potassium carbonate (0.42 g, 3.06 mmol), dioxane (5 mL), and water (1 mL) were added to a reaction flask. The mixture was stirred at 100°C for 16 hours. LCMS indicated the reaction was complete. The reaction mixture was concentrated, and the crude product was purified by column chromatography to afford A160-3 (0.5 g, 91%) as a white solid. m / z [M+H]+ =540.1.

[0430] 3) Synthesis of intermediate A160-4:

[0431] To a solution of A160-3 (0.5 g, 0.93 mmol) in tetrahydrofuran (5 mL) was added liquid bromine (0.74 g, 4.65 mmol). The mixture was stirred for 4 hours. LCMS indicated the reaction was complete. The reaction mixture was filtered to afford the yellow solid product A160-4 (0.5 g, 87% yield). m / z [M+H] + =617.2.

[0432] 4) Synthesis of intermediate A160-6:

[0433] A160-4 (0.2 g, 0.32 mmol), A160-5 (200 mg, 0.94 mmol), DIPA (0.6 g, 5.93 mmol), silver fluoroborate (0.2 g, 1.03 mmol), and DMSO (2 mL) were added to a reaction flask. The mixture was microwaved at 100°C for 12 hours. LCMS indicated the reaction was complete. The reaction mixture was diluted with dichloromethane (50 mL) and washed with water (50 mL). The organic phase was concentrated and the crude product was purified by column chromatography to afford A160-6 (65 mg, 27%) as a white solid. m / z [M+H] + =750.2.

[0434] 5) Synthesis of product A160:

[0435] Referring to the synthesis of compound A1 in Example 1, compound A160 was synthesized by replacing A1-5 with A160-6. m / z[M+H] + =785.4. 1 H NMR(400MHz, DMSO-d6)δ10.43(s,1H),9.00(dd,J=4.2,1.7Hz,1H),8.84–8.77(m,1H),8.59(d,J=7.4H z,2H),8.50(dt,J=8.8,2.5Hz,1H),8.20(d,J=8.9Hz,1H),7.65(dd,J=8.3,4.2Hz,1H),7.45(d,J=2.1H z,2H),5.37(s,2H),4.48(d,J=12.8Hz,1H),3.70(d,J=8.4Hz,1H),3.65-3.42(m,4H),3.10(ddd,J=30 .2,13.8,6.7Hz,3H),2.46(s,3H),1.69(d,J=7.6Hz,1H),1.40(q,J=7.5Hz,2H),1.27(d,J=7.0Hz,3H).

[0436] Example 59: Synthesis of Compound A159

[0437] Referring to the synthesis of compound A160 in Example 58, phenylboronic acid was used instead of 6-quinolineboronic acid pinacol ester to synthesize compound A159. m / z [MH] - =732.4.1H NMR(400MHz,DMSO-d6)δ10.41(s,1H),8.58(s,1H),8.14(dd,J=6.7,3.0Hz,2H ),7.56(dd,J=5.1,2.0Hz,3H),7.44(s,2H),5.32(s,2H),4.46(d,J=11.9Hz,1 H),3.67(t,J=8.8Hz,1H),3.48(s,2H),3.27(d,J=12.3Hz,2H),3.07(dt,J=29 .6,6.6Hz,3H),2.46(s,3H),1.67(d,J=7.8Hz,1H),1.39(s,2H),1.26(s,3H).

[0438] Example 61: Synthesis of Compound A162

[0439] Referring to the synthesis of compound A26 in Example 15, 6-(4-methylpiperazin-1-yl)pyridine-3-boronic acid pinacol ester was used instead of A26-1 to synthesize compound A162. m / z [MH] - =793.4. 1 H NMR(400MHz,DMSO-d6)δ10.36(s,1H),8.79(d,J=2.4Hz,1H),8.57(s,1H),8.19–8.08(m,1H ),7.40(s,2H),6.96(d,J=9.1Hz,1H),5.26(s,2H),4.53(d,J=12.3Hz,1H),3.60(t,J=5.1H z,4H),3.52(s,2H),3.26–3.23(m,2H),2.99(t,J=8.6Hz,3H),2.82(d,J=11.1Hz,1H),2.64 (d,J=11.0Hz,1H),2.44(s,3H),2.41(t,J=5.0Hz,4H),2.23(s,3H),1.21(t,J=7.5Hz,3H).

[0440] Example 62: Synthesis of Compound A163

[0441] Referring to the synthesis of compound A26 in Example 15, compound A163 was synthesized by replacing A36-1 with A145-1 and pyrazolo[1,5-a]pyridine-5-boronic acid pinacol ester with A26-1. m / z [MH] - =746.4. 1 H NMR (400MHz, DMSO-d6) δ10.40(s,1H),8.84(d,J=7.4Hz,1H),8.60(s,1H),8.46(d,J=1 .8Hz,1H),8.12(d,J=2.3Hz,1H),7.54(dd,J=7.3,1.9Hz,1H),7.45(s,2H),6.87(d,J= 2.2Hz,1H),5.34(s,2H),4.57(d,J=12.5Hz,1H),3.56(d,J=11.9Hz,4H),3.12–3.01(m ,3H),2.88(d,J=11.3Hz,1H),2.70(d,J=10.4Hz,1H),2.48(s,3H),1.28–1.24(m,3H).

[0442] Example 63: Synthesis of Compound A165

[0443] Referring to the synthesis of compound A26 in Example 15, 4-(4-(oxetan-3-yl)piperazin-1-yl)phenylboronic acid pinacol ester was used instead of A26-1 to synthesize compound A165. ESI (m / z) = 836.5 [M+H] + . 1 H NMR (400MHz, chloroform-d) δ9.25(s,1H),8.58(s,1H),8.49(d,J=8.7Hz,1H),8.18–8.11(m,2H),7.57(d,J=2.0Hz,1H), 7.51(dd,J=8.9,2.0Hz,1H),6.98–6.90(m,2H),5.66(d,J=12.8Hz,1H),5.16(s,2H),4.79(d,J=12.9Hz,1H),4.6 9(dt,J=17.5,6.3Hz,4H),3.90–3.76(m,2H),3.54(dt,J=17.9,9.0Hz,2H),3.36(t,J=5.0Hz,4H),3.19(t,J=7. 6Hz,2H),3.07(s,1H),2.80(dd,J=32.4,11.8Hz,2H),2.56(s,3H),2.50(t,J=5.0Hz,4H),1.34(t,J=7.5Hz,3H).

[0444] Example 64: Synthesis of Compound A166

[0445] Referring to the synthesis of compound A26 in Example 15, compound A166 was synthesized by replacing A36-1 with A145-1 and A26-1 with p-fluorophenylboronic acid. ESI (m / z) = 724.4 [MH] - . 1 H NMR(400MHz,DMSO-d6)δ10.36(s,1H),8.57(s,1H),8.24–8.10(m,2H),7.43–7.33(m,4H),5.29(s,2H),4.53(d,J=12.5Hz,1H),3.57– 3.47(m,3H),3.24(s,1H),3.01(p,J=8.6,7.8Hz,3H),2.84(d,J=11.2Hz,1H),2.69–2.62(m,1H),2.44(s,3H),1.22(t,J=7.4Hz,3H).

[0446] Example 65: Synthesis of Compound A167

[0447] Referring to the synthesis of compound A26 in Example 15, compound A167 was synthesized by replacing A36-1 with A145-1 and A26-1 with 2-methylpyridine-4-boronic acid. m / z [M+H] + =723.4. 1 H NMR (400MHz, DMSO-d6) δ10.41 (s, 1H), 8.65 (d, J = 5.1Hz, 1H), 8.60 (s, 1H), 7. 93(s,1H),7.85(d,J=5.1Hz,1H),7.44(s,2H),5.33(s,2H),4.56(d,J=12.5H z,1H),3.55(s,3H),3.30(s,2H),3.06(d,J=8.3Hz,2H),2.87(d,J=11.3Hz,1 H),2.70(d,J=10.7Hz,1H),2.60(s,3H),2.47(s,3H),1.26(d,J=7.2Hz,3H).

[0448] Example 66: Synthesis of Compound A168

[0449] Referring to the synthesis of compound A26 in Example 15, compound A168 was synthesized by replacing A36-1 with A145-1 and A26-1 with 1-methylpyrazole-4-boronic acid pinacol ester. m / z [M+H] +=712.4. 1 H NMR (400MHz, DMSO-d6) δ10.36(s,1H),8.55(s,1H),8.30(s,1H),7.91(s,1H),7.41(s,2H),5.24(s,2H),4.52(d,J=12.0Hz,1H),3.91(s ,3H),3.50(d,J=10.9Hz,4H),2.99(q,J=8.4Hz,3H),2.81(d,J=11.2Hz,1H),2.64(d,J=9.7Hz,1H),2.44(s,3H),1.19(d,J=7.4Hz,3H).

[0450] Example 67: Synthesis of Compound A174

[0451] Referring to the synthesis of compound A26 in Example 15, quinazoline-6-boronic acid pinacol ester was used instead of A26-1 to synthesize compound A174. m / z [M+H] + =748.4. 1 H NMR (400MHz, DMSO-d6) δ10.50(s,1H),9.80(s,1H),9.37(s,1H),8.93(s,1H),8.72(d,J=8. 6Hz,1H),8.59(s,1H),8.18(d,J=8.8Hz,1H),8.10(d,J=8.6Hz,1H),8.00(s,1H),7.73(d,J =8.8Hz,1H),5.47(s,2H),4.57(d,J=12.4Hz,1H),3.57(s,3H),3.31(s,2H),3.06(d,J=8.9 Hz,2H),2.89(d,J=11.4Hz,1H),2.72(d,J=11.1Hz,1H),2.47(s,3H),1.26(d,J=7.4Hz,3H).

[0452] Example 68: Synthesis of Compound A175

[0453] Referring to the synthesis of compound A160 in Example 58, compound A175 was synthesized by replacing A20-5 with A1-4. m / z[M+H] +=773.4.1H NMR(400MHz, DMSO-d6)δ10.48(s,1H),9.00(dd,J=4.2,1.7Hz,1H),8.79(d,J=1.9Hz,1H),8.62–8.54(m,2H),8.48 (dd,J=8.8,1.9Hz,1H),8.18(d,J=8.8Hz,1H),8.10(d,J=8.6Hz,1H),8.01(d,J=2.1Hz,1H),7.74(dd,J=8.8,2.1H z,1H),7.64(dd,J=8.3,4.2Hz,1H),5.47(s,2H),4.48(d,J=12.8Hz,1H),3.71(t,J=9.2Hz,1H),3.49(d,J=12.4Hz ,3H),3.09(dp,J=28.3,7.1Hz,4H),2.47(s,3H),1.70(d,J=7.9Hz,1H),1.52–1.34(m,2H),1.28(d,J=7.3Hz,3H).

[0454] Example 69: Synthesis of Compound A180

[0455] Referring to the synthesis of compound A26 in Example 15, compound A180 was synthesized by replacing A36-1 with A145-1 and pyridine-4-boronic acid with A26-1. m / z [M+H] + =709.4. 1 HNMR (400MHz, DMSO-d6) δ10.41(s,1H),8.76(d,J=5.1Hz,2H),8.56(s,1H),8.02(d,J=5.0Hz,2H),7.41(s,2H),5.31(s,2H),4.53(d,J=12.5Hz,1H ),3.51(d,J=11.5Hz,3H),3.27(s,1H),3.02(p,J=9.4,8.5Hz,3H),2.84( d,J=11.2Hz,1H),2.66(d,J=11.0Hz,1H),2.44(s,3H),1.24–1.20(m,3H).

[0456] Example 70: Synthesis of Compound A33

[0457] 1) Synthesis of intermediate A33-3:

[0458] Compound A33-1 (100 mg, 0.63 mmol), compound A33-2 (300 mg, 0.95 mmol), phosphoric acid (73 mg, 0.63 mmol), and ethanol (2 mL) were added to the reaction flask in sequence. The mixture was stirred at 80°C for 16 hours. LCMS indicated the reaction was complete. The reaction solution was filtered and washed with a mixture of ethyl acetate and petroleum ether to obtain the product A33-3 as an off-white solid (200 mg, yield: 75.18%). m / z [MH] - =422.4.

[0459] 2) Synthesis of intermediate A33-4:

[0460] Compound A33-3 (200 mg, 0.47 mmol), compound A20-5 (210 mg, 0.64 mmol), and N,N-diisopropylethylamine (0.18 g, 1.41 mmol) were dissolved in acetonitrile (3 mL) and stirred at 45°C for 16 hours. LCMS showed the reaction was complete. The reaction solution was filtered and washed with acetonitrile to give a white crude product A33-4 (0.12 g, yield: 37.86%). m / z [M+H] + =671.2.

[0461] 3) Synthesis of intermediate A33-5:

[0462] Compound A33-4 (0.12 g, 0.18 mmol) was dissolved in dichloromethane (2 mL) and trifluoroacetic acid (0.3 mL) was added dropwise. The reaction mixture was stirred at 25°C for 1 hour. LCMS showed that the reaction was complete. The reaction mixture was concentrated to give a yellow oil A33-5 (0.10 g, yield: 100%). m / z [M+H] + =571.2.

[0463] 4) Synthesis of product A33:

[0464] Compound A1-7 (30 mg, 0.19 mmol), N-hydroxy-7-azabenzotriazole (29 mg, 0.22 mmol), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (45 mg, 0.23 mmol) were dissolved in N,N-dimethylformamide (0.5 mL) and stirred at 25°C for 1 hour. Subsequently, compound A33-5 (0.1 g, 0.18 mmol) and N,N-diisopropylethylamine (70 mg, 0.54 mmol) were added to the reaction solution in sequence, and the reaction solution was stirred at 25°C for another 1 hour. LCMS showed the reaction was complete. The reaction solution was added to water, extracted with dichloromethane, washed with water and saturated brine, dried over sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (dichloromethane / ethyl acetate) to obtain a white product A33 (24.6 mg, yield: 19.86%, purity: 98.67%). m / z [MH] - =705.3. 1 H NMR (400MHz, DMSO-d6) δ10.34(s,1H),8.60(s,1H),8.02–7.89(m,2H),7.57–7.40(m,5H),6.89(s,1H),5.16(s,2H),4.56(d,J=12.7Hz,1H) ,3.65–3.51(m,3H),3.25(s,1H),3.01(d,J=8.8Hz,3H),2.85(d,J=11.2Hz,1H),2.67(d,J=11.1Hz,1H),2.47(s,3H),1.25(d,J=7.4Hz,3H).

[0465] Example 71: Synthesis of Compound A179

[0466] Referring to the synthesis of compound A160 in Example 58, 4-pyridine boronic acid was used instead of 6-quinoline boronic acid pinacol ester to synthesize compound A179. m / z [M+H] + =735.4. 1H NMR (400MHz, DMSO-d6) δ10.40(s,1H),8.79(d,J=5.2Hz,2H),8.58(s,1H),8.19–7.96(m,2H),7.43(s,2H),5.32(s,2H),4.45(d,J=12.4Hz,1H),3. 68(d,J=8.7Hz,1H),3.48(s,2H),3.20–2.97(m,4H),2.46(s,3H),2.02(d ,J=8.6Hz,1H),1.67(d,J=8.2Hz,1H),1.39(s,2H),1.26(d,J=6.5Hz,3H).

[0467] Example 72: Synthesis of Compound A164

[0468] Reference Example 58: Synthesis of Compound A160 using Compound A164 was synthesized by replacing 6-quinolineboronic acid pinacol ester. m / z[M+H] + =740.3. 1 H NMR (400MHz, DMSO-d6) δ10.37(s,1H),8.53(s,1H),7.76(d,J=4.3Hz,2H),7.41(t,J =5.7Hz,2H),7.22(t,J=4.4Hz,1H),5.25(s,2H),4.41(d,J=11.8Hz,1H),3.64(q,J= 8.8Hz,2H),3.46–3.42(m,2H),3.28-2.20(m,2H),3.11–3.05(m,1H),2.99(dd,J=14 .0,7.2Hz,1H),2.42(s,3H),1.63(q,J=7.8Hz,1H),1.37(m,2H),1.25-1.20(m,4H).

[0469] Example 73: Synthesis of Compound A169

[0470] Reference Example 70: Synthesis of Compound A33 using replace Compound A169 was synthesized. m / z[M+H] + =672.3. 1H NMR (400 MHz, chloroform-d) δ 11.78 (s, 1H), 8.76 (s, 1H), 8.58 (s, 1H), 7.93 (d, J = 8.9 Hz, 1H), 7.02–6.94 (m, 1H), 5.64 (d, J = 13.1 Hz, 1H), 5.04 (s, 2H), 4.77 (d, J = 12.8 Hz, 1H), 3.8 5–3.71(m,2H),3.47(s,1H),3.15(q,J=7.5Hz,2H),3.05(s,1H),2.85–2.67(m,2H), 2.56(s,3H),2.11(tt,J=8.2,5.1Hz,1H),1.30(t,J=7.5Hz,3H),1.13–1.00(m,4H).

[0471] Example 74: Synthesis of Compound A170

[0472] Reference Example 58: Synthesis of Compound A160 using Compound A170 was synthesized by replacing 6-quinolinylboronic acid pinacol ester. m / z[MH] - =789.4. 1 H NMR (400MHz, DMSO-d6) δ10.41(s,1H),9.54(s,1H),8.96(d,J=1.6Hz,1H),8.59(s,1H),8.32( dd,J=8.6,1.6Hz,1H),8.26(d,J=8.5Hz,1H),7.45(s,2H),5.35(s,2H),4.47(d,J=12.3Hz,1H ),3.50(s,1H),3.44(d,J=8.6Hz,2H),3.18–3.09(m,2H),3.05(t,J=7.0Hz,2H),2.47(s,3H), 1.68(d,J=7.2Hz,1H),1.40(dt,J=9.7,6.1Hz,2H),1.28(d,J=7.3Hz,3H),1.23–1.16(m,1H).

[0473] Example 75: Synthesis of Compound A176

[0474] Reference Example 58: Synthesis of Compound A160 using Compound A176 was synthesized by replacing 6-quinolinylboronic acid pinacol ester. m / z[M+H] + =749.4. 1H NMR(400MHz,DMSO-d6)δ10.40(s,1H),8.65(d,J=5.2Hz,1H),8.59(s,1H),7.95–7.91(m,1H ),7.84(d,J=5.3Hz,1H),7.44(s,2H),5.32(s,2H),4.45(d,J=11.8Hz,1H),3.71–3.63(m,1 H),3.49(s,2H),3.29(dd,J=13.8,6.4Hz,2H),3.08(ddd,J=39.0,13.9,7.2Hz,2H),2.60(s ,3H),2.46(s,3H),1.67(d,J=7.9Hz,1H),1.40(dd,J=21.9,10.3Hz,2H),1.29-1.15(m,4H).

[0475] Example 76: Synthesis of Compound A177

[0476] Reference Example 58: Synthesis of Compound A160 using Compound A177 was synthesized by replacing 6-quinolinylboronic acid pinacol ester. m / z[M+H] + =740.4. 1 H NMR(400MHz,DMSO-d6)δ10.36(s,1H),8.59(s,1H),7.53–7.39(m,2H),6.96–6.83(m,1H) ,5.24(s,2H),4.44(d,J=11.5Hz,1H),4.30(q,J=2.8Hz,2H),3.84(t,J=5.4Hz,2H),3.65 (t,J=8.8Hz,1H),3.52–3.40(m,2H),3.27(d,J=10.4Hz,2H),3.04(dp,J=28.6,7.1Hz,2H ),2.59-2.50(m,2H),2.46(s,3H),1.65(d,J=7.4Hz,1H),1.38(s,2H),1.26-1.20(m,4H).

[0477] Example 77: Synthesis of Compound A182

[0478] 1) Synthesis of intermediate A182-3:

[0479] To a 250 mL single-necked flask, A182-1 (8 g, 49.64 mmol), acetic acid (80 mL), and A182-2 (9.69 g, 74.46 mmol) were added sequentially and stirred at 90°C for 16 hours. The reaction mixture was returned to room temperature, filtered, and the solid was concentrated to dryness to obtain the product A182-3 (10 g, yield: 83.5%). ESI (m / z) = 242.2 [M+H] + .

[0480] 2) Synthesis of intermediate A182-4:

[0481] To a 250 mL single-necked flask, A182-3 (10 g, 41.45 mmol), acetonitrile (80 mL), and N,N-diisopropylethylamine (16.1 g, 124.35 mmol) were added sequentially. A20-5 (14.3 g, 43.52 mmol) was slowly added while stirring at room temperature. The mixture was stirred at 45°C for 16 hours. The reaction mixture was returned to room temperature, filtered, and the solid was concentrated to dryness to obtain the product A182-4 (16.4 g, yield: 81%). ESI (m / z) = 489.2 [M+H] + .

[0482] 3) Synthesis of A182

[0483] Referring to the synthesis of compound A160 in Example 58, compound A182 was synthesized by replacing A160-3 with A182-4. m / z[M+H] + =735.3. 1 H NMR (400MHz, DMSO-d6) δ10.38(s,1H),8.73(d,J=4.8Hz,1H),8.58(d,J=18.1Hz,1H),8.17(d,J=7.9 Hz,1H),7.98(td,J=7.7,1.8Hz,1H),7.53(dd,J=7.6,4.9Hz,1H),7.47–7.38(m,2H),5.31(s,2H),4. 43(d,J=12.7Hz,1H),3.73–3.58(m,1H),3.54–3.37(m,3H),3.27(dd,J=9.8,5.1Hz,1H),3.13–3.05( m,1H),3.03–2.95(m,1H),2.43(s,3H),1.65(d,J=7.4Hz,1H),1.48–1.29(m,2H),1.26–1.20(m,4H).

[0484] Example 78: Synthesis of Compound A181

[0485] 1) Synthesis of intermediate A181-2:

[0486] Compound A181-1 (50 g, 342 mmol) was added to acetonitrile (500 mL), and then a 2 mol / L tetrahydrofuran solution of dimethylamine (200 mL) was added. o The reaction was allowed to proceed for 4 hours. Hydrazine hydrate (51.35 g, 820.6 mmol) was then added and the reaction was refluxed overnight. LCMS indicated that the reaction was complete. The reaction solution was returned to room temperature and concentrated under reduced pressure (remaining 230 mL). Solid precipitated and was filtered to obtain a filter cake. The filter cake was dried to obtain the product (39 g, yield: 89.7%). m / z [M+H] + =128.0.

[0487] 2) Synthesis of A181:

[0488] Referring to the synthesis of compound A182 in Example 77, compound A181 was synthesized by replacing A182-1 with A181-2. m / z [M+H] + =701.4. 1 H NMR (400MHz, CDCl3) δ11.66(s,1H),8.83(s,1H),8.59(s,1H),7.95(d,J=8.8 Hz,1H),6.98(d,J=8.9Hz,1H),4.98(s,2H),4.85-4.70(m,0.5H),4.30-4.00(m,2H),3.95-3.55(m,1.5H),3.23(dd,J=52.7,10.8Hz,2 H),3.11(s,6H),3.09-3.00(m,1H),2.54(s,3H),2.34(d,J=26.2Hz,1H),1.80-1.70(m,1H),1.55-1.35(s,2H),1.28(t,J=7.5Hz,4H).

[0489] Example 79: Synthesis of Compound A183

[0490] Reference Example 77 Synthesis of Compound A182 using replace Compound A183 was synthesized. m / z[M+H] + =709.1. 1H NMR (400MHz, DMSO-d6) δ10.38(s,1H),8.74–8.70(m,1H),8.58(s,1H),8.18(dt,J=7.9,1.1H z,1H),7.98(td,J=7.8,1.8Hz,1H),7.53(ddd,J=7.6,4.8,1.2Hz,1H),7.47–7.38(m,2H),5. 32(s,2H),4.54(d,J=12.5Hz,1H),3.52(t,J=10.3Hz,3H),3.29–3.21(m,1H),3.02(d,J=9.6 Hz,3H),2.85(d,J=11.3Hz,1H),2.67(d,J=10.4Hz,1H),2.45(s,3H),1.22(t,J=7.4Hz,3H).

[0491] Example 80: Synthesis of Compound A184

[0492] Referring to the synthesis of compound A160 in Example 58, 3-pyridine boronic acid was used instead of 6-quinoline boronic acid pinacol ester to synthesize compound A184. m / z [M+H] + =735.4. 1 H NMR (400MHz, DMSO-d6) δ10.42(s,1H),9.30(s,1H),8.75(d,J=4.8Hz,1H),8.60(s,1H),8.46( dd,J=8.0,2.0Hz,1H),7.62(dd,J=7.9,4.8Hz,1H),7.45(d,J=4.2Hz,2H),5.33(s,2H),4.46(d ,J=12.1Hz,1H),3.69(q,J=11.8,9.4Hz,1H),3.49(s,4H),3.20–3.09(m,1H),3.08–2.97(m,1H ), 2.47 (s, 3H), 1.68 (d, J = 8.0Hz, 1H), 1.40 (q, J = 7.7, 7.2Hz, 2H), 1.26 (td, J = 6.8, 3.6Hz, 4H).

[0493] Example 81: Synthesis of Compound A187

[0494] 1) Synthesis of intermediate A187-1:

[0495] To a 100 mL single-necked flask, A182-2 (5 g, 34.68 mmol), dichloromethane (50 mL), and sulfonyl chloride (4.91 g, 36.41 mmol) were added sequentially and stirred at 25°C for 16 hours. The reaction mixture was extracted three times with water and DCM. The organic phase was dried over anhydrous sodium sulfate and concentrated to afford product 187-1 (6 g, yield: 96.86%).

[0496] 2) Synthesis of intermediate A187-2:

[0497] A187-1 (6 g, 33.59 mmol), acetonitrile (60 mL), (7.13 g, 33.59 mmol) and triethylamine (11.9 g, 117.57 mmol) were stirred at 25°C for 16 hours. The reaction mixture was filtered, the filtrate was concentrated, and silica gel column chromatography gave the product A187-2 (9.2 g, yield: 80.45%). ESI (m / z) = 341.3 [M+H] + .

[0498] 3) Synthesis of intermediate A187-3:

[0499] A187-2 (3 g, 18.41 mmol), ethanol (30 mL), (7.18 g, 20.25 mmol) and phosphoric acid (9.02 g, 92.05 mmol) were stirred at 80°C for 48 hours. The reaction solution was returned to room temperature and filtered to obtain the product A187-3 (3 g, yield: 35.95%). ESI (m / z) = 353.2 [M+H] + .

[0500] 4) Synthesis of intermediate A187-4:

[0501] To a 100 mL single-necked flask, A187-3 (2.87 g, 8.13 mmol), tetrahydrofuran (30 mL), di-tert-butyl dicarbonate (2.66 g, 12.20 mmol), and N,N-diisopropylethylamine (3.15 g, 24.39 mmol) were added sequentially and stirred at room temperature for 15 hours. The reaction mixture was concentrated, extracted, and purified by silica gel column chromatography to afford the product A187-4 (2.4 g, yield: 65.16%). ESI (m / z) = 452.3 [MH] - .

[0502] 5) Synthesis of intermediate A187-5:

[0503] To a 100 mL single-necked flask, A187-4 (2.04 g, 4.5 mmol), acetonitrile (20 mL), A20-5 (1.48 g, 4.5 mmol), and N,N-diisopropylethylamine (1.74 g, 13.5 mmol) were added sequentially and stirred at 45°C for 7 hours. The reaction mixture was concentrated and purified by silica gel column chromatography to afford the product A187-5 (0.23 g, yield: 63.41%). ESI (m / z) = 699.3 [MH] - .

[0504] 6) Synthesis of intermediate A187-6:

[0505] To a 10 mL single-necked flask, A187-5 (0.26 g, 0.33 mmol), acetonitrile (2 mL), tetrahydropyrrole (0.12 g, 1.65 mmol), and N,N-diisopropylethylamine (0.22 g, 1.65 mmol) were added sequentially and stirred at 80°C for 18 hours. The reaction mixture was concentrated and purified by silica gel column chromatography to afford the product A187-6 (0.23 g, yield: 99.68%). ESI (m / z) = 691.4 [MH] - .

[0506] 7) Synthesis of intermediate A187-7:

[0507] A187-6 (0.23 g, 0.33 mmol), dichloromethane (5 mL), and trifluoroacetic acid (2 mL) were added sequentially to a 10 mL single-necked flask and stirred at room temperature for 1 hour. The reaction mixture was concentrated to afford the product A187-7 (0.23 g, yield: 99.6%). ESI (m / z) = 591.3 [MH] - .

[0508] 8) Synthesis of A187:

[0509] Add the following into a 10mL single-necked bottle: (90 mg, 0.55 mmol), N-hydroxy-7-azabenzotriazole (80 mg, 0.58 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (110 mg, 0.58 mmol) were dissolved in N,N-dimethylformamide (0.5 mL) and stirred at 25°C for 1 hour. A187-7 (190 mg, 0.32 mmol) and N,N-diisopropylethylamine (120 mg, 0.96 mmol) were then added to the reaction solution in sequence, and the reaction solution was stirred at 25°C for another 2 hours. LCMS showed that the reaction was complete, and the reaction solution was filtered to obtain a filtrate, which was purified by reverse phase column chromatography (0.05% aqueous formic acid / acetonitrile) to obtain compound A187. m / z [M+H] + =727.3.1 H NMR(400MHz,DMSO-d6)δ10.29(s,2H),8.55(s,1H),7.46–7.36(m,2H),5.1 1(s,2H),4.40(d,J=11.5Hz,1H),3.63(q,J=9.0Hz,1H),3.45–3.35(m,7H), 3.21(d,J=10.6Hz,1H),2.95(dp,J=21.5,7.2Hz,2H),2.43(s,3H),1.92–1. 87(m,4H),1.60(q,J=7.6Hz,1H),1.40–1.28(m,2H),1.17(t,J=7.4Hz,4H).

[0510] Example 82: Synthesis of Compound A185

[0511] Reference Example 81 Synthesis of Compound A187 using Compound A185 was synthesized by replacing tetrahydropyrrole. m / z[M+H] + =789.1. 1 H NMR (400MHz, DMSO_d6): δ10.29-10.28(m,2H),8.53(s,1H),7.43-7.36(m,2H), 5.10(s,2H),4.40-4.38(m,1H),3.62-3.60(m,1H),3.53-3.50(m,2H),3.42-3.35(m,2H),3.21-3.19(m,1H),3. 00-2.91(m,2H),2.45-2.42(m,6H),2.21-2.15(m,4H),1.60-1.59(m,1H),1.39-1.28(m,2H),1.19-1.14(m,4H).

[0512] Example 83: Synthesis of Compound A186

[0513] Reference Example 70: Synthesis of Compound A33 using Compound A186 was synthesized by replacing A33-1 with A187-2. m / z[M+H] + =743.1. 1H NMR (400MHz, DMSO_d6): δ10.34(s,1H),8.57(s,1H),7.54–7.37(m,2H),5.13(s,2H),4.42(d,J=11.7Hz,1H),3.70(t,J=4.7Hz,4H),3.64( d,J=8.0Hz,2H),3.48-3.38(m,5H),3.23(d,J=11.4Hz,1H),3.09–2.87(m,2H),2.45(s,3H),2.12–1.89(m,1H),1.62(d,J=7.0Hz,1H),1.38 -1.30(m,2H),1.21-1.13(m,4H).

[0514] Example 84: Synthesis of Compound A188

[0515] Referring to the synthesis of compound A187 in Example 81, (S)-2-methylpyrrolidine was used instead of tetrahydropyrrole to synthesize compound A188. m / z [MH] - =739.4. 1 H NMR(400MHz,DMSO-d6)δ10.30(s,1H),8.55(s,1H),7.48–7.36(m,2H),5.10(s,2H) ),4.40(d,J=11.4Hz,1H),3.92(td,J=6.7,3.5Hz,1H),3.62(d,J=8.2Hz,1H),3.44 (dd,J=8.8,4.1Hz,4H),3.21(d,J=11.0Hz,1H),2.96(q,J=10.4,8.5Hz,2H),2.42 (s,3H),2.10–1.77(m,4H),1.61(d,J=8.6Hz,2H),1.33(s,2H),1.24–1.13(m,7H).

[0516] Example 85: Synthesis of Compound A189

[0517] Referring to the synthesis of compound A187 in Example 81, (R)-2-methylpyrrolidine was used instead of tetrahydropyrrole to synthesize compound A189. m / z [MH] - =739.4. 1H NMR(400MHz,DMSO-d6)δ10.30(s,1H),8.55(s,1H),7.45–7.35(m,2H),5.10(s,2H), 4.40(d,J=11.3Hz,1H),3.92(dq,J=7.0,3.7Hz,1H),3.68–3.58(m,1H),3.47–3.35(m ,4H),3.20(d,J=10.2Hz,1H),2.95(dp,J=28.2,7.0Hz,2H),2.43(s,3H),2.13–1.78 (m,4H),1.61(dq,J=10.6,6.3,5.6Hz,2H),1.33(t,J=6.7Hz,2H),1.22–1.13(m,7H).

[0518] Example 86: Synthesis of Compound A190

[0519] Reference Example 81 Synthesis of Compound A187 using Compound A190 was synthesized by replacing tetrahydropyrrole. m / z[M+H] + =782.4. 1 H NMR (400MHz, chloroform-d) δ8.85(s,1H),8.59(s,1H),7.99(d,J=8.9Hz,1H),6.99(d,J=8.9Hz,1H),4.97(s,2H),4.84–3.77(m,3H),3.59(t,J=5.1H z,5H),3.39–3.01(m,3H),2.70(t,J=5.0Hz,4H),2.54(s,3H),2.41–1.76(m,2H),1.27(dd,J=13.7,6.1Hz,7H),0.48(dd,J=9.3,4.9Hz,4H).

[0520] Example 87: Synthesis of Compound A191

[0521] Reference Example 81 Synthesis of Compound A187 using Compound A191 was synthesized by replacing tetrahydropyrrole. m / z[M+H] + =796.4. 1H NMR (400MHz, chloroform-d) δ8.87(s,1H),8.59(s,1H),7.96(d,J=8.9Hz,1H),6.98(d,J=8.9Hz,1 H),4.98(s,2H),4.83–3.98(m,3H),3.67(t,J=5.0Hz,6H),3.47–2.98(m,3H),2.66(t,J= 5.0Hz,4H),2.53(s,3H),2.35(d,J=6.6Hz,2H),1.88(d,J=117.4Hz,2H),1.45(s,1H),1. 34–1.21(m,4H),0.89(dt,J=13.0,6.5Hz,1H),0.59–0.52(m,2H),0.13(t,J=5.2Hz,2H).

[0522] Example 88: Synthesis of Compound A192

[0523] Reference Example 81 Synthesis of Compound A187 using Compound A192 was synthesized by replacing tetrahydropyrrole. m / z[M+H] + =715.4. 1 H NMR(400MHz,DMSO-d6)δ10.31(s,1H),8.52(s,1H),7.47–7.33(m,2H),5.0 9(s,2H),4.40(d,J=11.1Hz,1H),3.62(d,J=8.5Hz,1H),3.43(p,J=6.2,5.3 Hz,4H),3.21(d,J=9.0Hz,2H),2.94(s,5H),2.42(s,3H),1.60(d,J=6.2Hz ,1H),1.28(d,J=40.1Hz,2H),1.17(t,J=7.4Hz,4H),1.07(t,J=7.0Hz,3H).

[0524] Example 89: Synthesis of Compound A193

[0525] Referring to the synthesis of compound A187 in Example 89, azetidine was used instead of tetrahydropyrrole to synthesize compound A193. m / z [MH] - =711.4. 1H NMR (400MHz, DMSO-d6) δ10.30(d,J=14.4Hz,2H),8.55(s,1H),7.41(d,J=3.6Hz,2H),5.1 0(s,2H),4.40(d,J=11.2Hz,1H),3.96(t,J=7.5Hz,5H),3.61(d,J=9.0Hz,1H),3.41(d,J= 9.5Hz,1H),3.20(d,J=11.0Hz,1H),3.02–2.95(m,1H),2.94–2.85(m,1H),2.43(s,3H),2 .36–2.30(m,2H),1.60(d,J=6.6Hz,1H),1.33(s,2H),1.23(s,1H),1.16(t,J=7.4Hz,4H).

[0526] Example 90: Synthesis of Compound A194

[0527] Referring to the synthesis of compound A187 in Example 81, 3-methoxy-azetidine was used instead of tetrahydropyrrole to synthesize compound A194. m / z [MH] - =741.4. 1 H NMR (400MHz, DMSO-d6) δ10.28(s,1H),8.55(s,1H),7.45–7.36(m,2H),5.10(s,2H),4.40(d,J=1 1.5Hz,1H),4.31(tt,J=6.1,4.3Hz,1H),4.17(dd,J=8.8,6.2Hz,2H),3.79(dd,J=9.0,4.2Hz,2H) ,3.65–3.56(m,1H),3.41(d,J=8.9Hz,2H),3.22(s,3H),3.19(s,1H),3.04–2.86(m,2H),2.42(s ,3H),1.60(d,J=6.8Hz,1H),1.33(d,J=6.7Hz,2H),1.22(d,J=5.3Hz,1H),1.16(t,J=7.4Hz,4H).

[0528] Example 91: Synthesis of Compound A195

[0529] Reference Example 81 Synthesis of Compound A187 using Compound A195 was synthesized by replacing tetrahydropyrrole. m / z[M+H] + =749.0. 1H NMR (400MHz, CDCl3): δ8.53 (s, 1H), 8.48 (s, 1H), 7.90 (d, J = 8.8Hz, 1H), 6. 93(d,J=8.8Hz,1H),5.44-4.92(m,2H),4.64-4.70(m,1H),4.44-4.38(m,4H ),4.14-3.97(m,2H),3.79-3.61(m,2H),3.26-2.98(m,2H),2.47(s,3H),2 .29-2.26(m,1H),2.15-2.10(m,1H),1.98-1.71(m,2H),1.24-1.18(m,4H).

[0530] Example 92: Synthesis of Compound A196

[0531] Reference Example 81 Synthesis of Compound A187 using Compound A196 was synthesized by replacing tetrahydropyrrole. m / z[M+H] + =757.0. 1 H NMR (400MHz, DMSO_d6): δ10.32(s,1H),8.50(brs,1H),7.42(q,J=8.4Hz,2H), 5.11(s,2H),4.44-4.38(m,1H),4.06-4.01(m,1H),3.65-3.58(m,1H),3.49-3 .37(m,5H),3.30-3.18(m,3H),3.23(s,3H),3.01-2.88(m,2H),2.41(s,3H),2 .06-1.95(m,2H),1.62-1.57(m,1H),1.37-1.29(m,2H),1.17(t,J=7.2Hz,4H).

[0532] Example 93: Synthesis of Compound A197

[0533] Reference Example 81 Synthesis of Compound A187 using Compound A197 was synthesized by replacing tetrahydropyrrole. m / z[M+H] + =757.0. 1H NMR (400MHz, DMSO_d6): δ10.28(s,2H),8.51(s,1H),7.43-7.37(m,2H),5. 10(s,2H),4.39(d,J=11.6Hz,1H),4.04-4.02(m,1H),3.65-3.58(m,1H),3 .48-3.37(m,5H),3.33-3.18(m,6H),3.01-2.88(m,2H),2.42(s,3H),2.03 -1.98(m,2H),1.61-1.56(m,1H),1.34-1.31(m,2H),1.16(d,J=7.6Hz,4H).

[0534] Example 94: Synthesis of Compound A198

[0535] Reference Example 81 Synthesis of Compound A187 using Compound A198 was synthesized by replacing tetrahydropyrrole. m / z[M+H] + =769.1. 1 H NMR (400MHz, CDCl3): δ8.79 (s, 1H), 8.52 (s, 1H), 7.89 (d, J = 9.2Hz, 1H), 6.91 (d,J=9.2Hz,1H),4.88(s,2H),4.68(s,1H),4.14-4.12(m,3H),4.02-3.96(m ,3H),3.82-3.79(m,3H),3.62-3.58(m,1H),3.25-2.99(m,4H),2.47(s,3H), 2.26-2.23(m,1H),2.10-2.07(m,2H),1.97-1.52(m,4H),1.23-1.18(m,3H).

[0536] Example 95: Synthesis of Compound A201

[0537] Reference Example 59: Synthesis of Compound A159 using replace Compound A201 was synthesized. m / z[M+H] + =734.3. 1H NMR (400MHz, DMSO_d6): δ10.38(d,J=12.0Hz,2H),8.59(s,1H),8.14(dd,J=6.7,3.0Hz,2H),7.56(dd,J=5.1,2.0Hz,3H),7.43(s,2H),5.31(s,2H),4. 47(s,1H),3.69(d,J=8.6Hz,1H),3.47(d,J=7.4Hz,2H),3.29–2.94(m,4H) ,2.46(s,3H),1.67(d,J=7.7Hz,1H),1.40-1.33(m,2H),1.29-1.23(m,4H).

[0538] Example 96: Synthesis of Compound A202

[0539] Reference Example 58: Synthesis of Compound A160 using Compound A202 was synthesized by replacing 6-quinolineboronic acid pinacol ester. m / z[M+H] + =724.0. 1 H NMR (400MHz, DMSO_d6): δ10.36(s,1H),8.52(s,1H),7.93(s,1H),7.42(s, 2H),7.15(d,J=3.6Hz,1H),6.70(brs,1H),5.26(s,2H),4.43-4.40(m,1H), 3.66-3.59(m,1H),3.51-3.40(m,2H),3.10-2.95(m,4H),2.67-2.59(m,1H) ,2.42(s,3H),1.65-1.60(m,1H),1.38-1.34(m,2H),1.22(t,J=7.2Hz,4H).

[0540] Example 97: Synthesis of Compound A199

[0541] Reference Example 58: Synthesis of Compound A160 using Replace 6-quinolinylboronic acid pinacol ester and use Compound A199 was synthesized instead of A1-7. m / z[M+H] + =725.4. 1H NMR(400MHz,DMSO-d6)δ 10.32(s,1H),8.04(s,1H),7.40(d,J=2.9Hz,2H),7.27(d,J=2.8Hz,2H),6.89–6 .80(m,1H),5.20(s,2H),4.41(d,J=9.4Hz,1H),4.26(q,J=2.8Hz,2H),3.81(t,J= 5.4Hz,2H),3.58(d,J=8.6Hz,1H),3.41(d,J=10.6Hz,2H),3.19(d,J=9.4Hz,1H), 3.01(dp,J=29.3,7.1Hz,2H),2.51(s,3H),1.82–1.52(m,1H),1.40–1.05(m,6H).

[0542] Example 98: Synthesis of Compound A200

[0543] Reference Example 58: Synthesis of Compound A160 using Replace 6-quinolinylboronic acid pinacol ester and use Compound A200 was synthesized instead of A1-7. m / z[M+H] + =724.4. 1 H NMR(400MHz, DMSO_d6)δ10.31(s,1H),9.80(s,1H),7.40(d,J=2.7Hz,2H),7.26–7.14(m,2H),6.83 (q,J=7.4Hz,3H),5.20(s,2H),4.26(q,J=2.8Hz,3H),3.81(t,J=5.4Hz,2H),3.58(d,J=8.6Hz,1H), 3.38(d,J=12.6Hz,3H),3.27(d,J=13.2Hz,3H),3.15(d,J=11.6Hz,1H),3.05(dd,J=13.8,7.2Hz,1 H),2.99–2.90(m,1H),1.66–1.57(m,1H),1.34(dd,J=11.9,4.6Hz,1H),1.21(q,J=7.5,6.5Hz,4H).

[0544] Biological testing section

[0545] Experimental Example 1 Biological Activity Test

[0546] ATPase activity assay of WRN helicase

[0547] The commercial ADP-Glo ​​assay kit (Promega, #V9102) was used to detect the ADP content produced by the hydrolysis of ATP by WRN helicase, which can reflect the ATPase activity of WRN helicase.

[0548] 45 nt oligo DNA single strand FLAP26 (TTTTTTTTTTTTTTTTTTTTTTCCAAGTAAAACGACGGCCAGTGC) was synthesized by Ascent Biotechnology Co., Ltd. See, for example, Brosh RM Jr et al., J Biol Chem, 2002 Jun; 277(26): 23236-45. Prepare reaction buffer (30 mM Tris pH 7.5, 2 mM MgCl2, 0.02% BSA, 50 mM NaCl, 0.1% pluronic F127) and sequentially add 5 μL of 3× test compound (diluted to 0.5% DMSO in reaction buffer, with a final starting concentration of 10 μM, and diluted 1:3 in a total of nine steps) to a 384-well clear plate. Mix thoroughly by shaking and incubate at 37°C for 3 hours. Next, prepare a 3× FLAP26 solution in reaction buffer and add 5 μL of this solution (final FLAP26 concentration of 0.4 nM) to the 384-well clear plate. Mix thoroughly by shaking to initiate the enzymatic reaction and incubate at room temperature for 30 minutes.

[0549] Transfer 5 μL of the above mixture to a 384-well white plate, add 5 μL of ADP-Glo ​​reagent, shake and mix, and incubate at room temperature in the dark for 40 minutes. Add 10 μL of kinase detection reagent to the above solution, shake and mix, incubate at room temperature in the dark for 30 minutes, and record the chemiluminescence reading. Calculate the inhibition rate of the compound on enzyme activity, and use nonlinear regression (dose response-variable slope) to fit the inhibition rate value and the logarithm of the compound concentration to obtain the IC value of the test compound. 50 value.

[0550] Table 1 WRN ATPase activity data of the compounds in the examples of this application

[0551] As can be seen from Table 1, the compounds of the present application have good inhibitory activity on the ATPase activity of the WRN protein.

[0552] Tumor cell proliferation inhibitory activity assay

[0553] The WRN gene was stably knocked out in DLD1 cells using CRISPR / Cas9 technology to construct the DLD1-WRN-KO cell line, which was used to evaluate the potential off-target effects of the compounds.

[0554] Human colon adenocarcinoma SW48 cells were cultured as monolayers in DMEM supplemented with 10% fetal bovine serum, 1% penicillin, and streptomycin at 37°C with 5% CO2. Human colon adenocarcinoma HCT116 cells were cultured as monolayers in McCoy's 5A medium supplemented with 10% fetal bovine serum, 1% penicillin, and streptomycin at 37°C with 5% CO2. DLD1-WRN-KO cells were cultured as monolayers in 1640 medium supplemented with 10% fetal bovine serum, 1% penicillin, and streptomycin at 37°C with 5% CO2. Cells were passaged twice weekly using trypsinization. Microsatellite-instable SW48 and HCT116 cells, as well as control DLD1-WRN-KO cells, were treated with test compounds for 4 days. ATP levels were then measured using the Novozymes CellCounting-Lite kit to assess the inhibitory effects of the test compounds on tumor cell growth.

[0555] In the present application, SW48 cell lines, HCT116 cell lines and DLD1-WRN-KO cell lines were seeded into 96-well cell culture plates at an appropriate cell density. After 24 hours, the test compound was used to treat the cells with a maximum concentration of 10 μM and 9 gradient dilutions of 1:3. A DMSO treatment group was also set up. Cultured in a 37°C / 5% CO2 incubator for 4 days. In order to test the inhibition of tumor cell proliferation by the test compound, the cells were equilibrated at room temperature for 30 minutes, and then 100 μL of cell proliferation detection reagent CellCounting-Lite (CCL) was added to each well. After shaking for 5 minutes, the cells were incubated in the dark for 10 minutes. The chemiluminescence value was read using a Thermo Varioskan LUX-3020 multifunctional microplate reader to convert it into a proliferation index to calculate the inhibition rate of the compound on tumor cell proliferation. The inhibition rate value and the logarithm of the compound concentration were fitted using nonlinear regression (dose response-variable slope) to obtain the IC value of the compound. 50 value.

[0556] Table 2 Tumor cell proliferation inhibitory activity data of the compounds in the examples of this application

[0557] As can be seen from Table 2, the compounds of the present application have good proliferation inhibitory activity against microsatellite unstable SW48 cells and HCT116 cells, but have no significant proliferation inhibitory activity against WRN knockout DLD1 cells, and have good selectivity.

[0558] Experimental Example 2 Liver microsome stability test

[0559] The test compounds were assessed for their first-phase metabolic stability in liver microsomes of CD-1 mice, Sprague-Dawley rats, beagle dogs, cynomolgus monkeys, and humans.

[0560] Experimental system:

[0561] The animal and human liver microsomes used in this test system were purchased from Xenotech, Corning or other qualified suppliers and stored in a freezer below -60°C before use.

[0562] Experiment Introduction:

[0563] Test and control compounds were incubated with animal and human liver microsomes at 37 ± 1°C for a specified period of time, with a maximum incubation time of 60 minutes. Samples were removed at designated time points and the reaction was terminated with acetonitrile or other organic solvent containing an internal standard. After centrifugation, the resulting supernatant was analyzed by liquid chromatography-tandem mass spectrometry (LC-MS / MS).

[0564] Experimental methods:

[0565] 1. Preparation of Buffer

[0566] Dissolve 73.21 g of potassium phosphate dibasic trihydrate and 10.78 g of potassium dihydrogen phosphate in 4000 mL of ultrapure water. Adjust the pH of the solution to 7.40 ± 0.10 using 10% phosphoric acid or 1 M potassium hydroxide, for a final concentration of 100 mM.

[0567] 2. Preparation of working solution

[0568] The test sample powder is prepared into a stock solution of a certain concentration using DMSO or other organic solvents, and then further diluted with a suitable organic solvent.

[0569] The control compounds testosterone, diclofenac, and propafenone were prepared as 10 mM stock solutions in DMSO and then further diluted in appropriate organic solvents.

[0570] 3. Preparation of Liver Microsome Solution

[0571] Dilute each microsome to a 2x working solution using 100 mM potassium phosphate buffer. The final concentration of microsomes in the reaction system is 0.5 mg / mL.

[0572] 4. Preparation of reduced nicotinamide adenine dinucleotide phosphate (NADPH) regeneration system

[0573] Weigh an appropriate amount of nicotinamide adenine dinucleotide phosphate (NADP) and isocitrate (ISO) powder, dissolve in magnesium chloride solution, and vortex to mix thoroughly. Add an appropriate amount of isocitrate dehydrogenase (IDH) and gently invert the solution to mix thoroughly. The final concentrations in the reaction system are: 1 mM NADP, 1 mM magnesium chloride, 6 mM ISO, and 1 unit / mL IDH.

[0574] 5. Preparation of Stop Solution

[0575] The stop solution is prepared with acetonitrile or other organic solvent containing an internal standard (tolbutamide or other suitable compound). The prepared stop solution is stored in a refrigerator at 2-8°C.

[0576] 6. Incubation Process

[0577] Incubations will be performed in 96-well plates. Prepare eight incubation plates, designated T0, T5, T15, T30, T45, T60, Blank60, and NCF60. The first six plates correspond to reaction time points of 0, 5, 15, 30, 45, and 60 minutes, respectively. In the Blank60 plate, no test or control compound is added, and a sample is taken after 60 minutes of incubation. In the NCF60 plate, potassium phosphate buffer is used instead of the NADPH regeneration system solution and incubated for 60 minutes. All conditions are replicated in triplicate.

[0578] Mix the microsomes with the test article or control compound, then preincubate the Blank60, T5, T15, T30, T45, and T60 plates (excluding T0 and NCF60) in a 37°C waterbath for approximately 10 minutes. Add the stop solution to the T0 plate, followed by the NADPH regeneration system working solution. In the NCF60 plate, 98 μL of potassium phosphate buffer was added to each well to initiate the reaction. After the preincubation period, 98 μL of the NADPH regeneration system working solution was added to each well of the Blank60, T5, T15, T30, T45, and T60 plates to initiate the reaction. The reaction temperature was 37 ± 1°C, and the final reaction volume was 200 μL. The reaction system contained 0.5 mg / mL microsomes, 1.0 μM substrate, 1 mM NADP, 6 mM ISO, and 1 unit / mL IDH.

[0579] The reaction was terminated by adding cold stop solution containing internal standard to the reaction plate at 5, 15, 30, 45 and 60 minutes, respectively.

[0580] All reaction plates were shaken and centrifuged at 3220 × g for 20 minutes at 4°C. The supernatant was diluted to a certain ratio and then analyzed by LC-MS / MS.

[0581] Sample analysis

[0582] Sample analysis was performed using liquid chromatography-tandem mass spectrometry (LC-MS / MS) without a standard curve or quality control samples. Semiquantitative determination was performed using the ratio of the analyte peak area to the internal standard peak area. Analyst software (Sciex, Framingham, Massachusetts, USA) was used for analyte and internal standard retention times, chromatogram acquisition, and chromatogram integration.

[0583] The CV of the internal standard peak area in each matrix should be within 20% for each analytical run.

[0584] Data Analysis

[0585] The in vitro elimination rate constant ke of the compound was obtained by converting the ratio of the compound to the internal standard peak area into a residual rate using the following formula:

[0586] CL int(mic) =0.693 / T 1 / 2 / Microsomal protein content (microsomal concentration during incubation mg / mL)

[0587] CL int(liver) =CL int(mic) × amount of microsomal protein in the liver (mg / g) × liver weight to body weight ratio

[0588] According to the well stir model, the hepatic intrinsic clearance and hepatic clearance can be converted using the following formula.

[0589] CL (liver) =(CL int(liver) *Q h ) / (CL int(liver) +Q h )

[0590] The parameters in the formula are shown in the table below.

[0591] Parameters in data analysis formulas

[0592] Experimental Example 3 Hepatocyte Metabolic Stability Test

[0593] This experiment was used to test the metabolic stability of compounds in hepatocytes.

[0594] Prepare 0.5 x 10 6 / mL of hepatocyte suspension, then 198μL of preheated cell suspension was added to a 96-well plate. 2μL of the test compound was added to each well of the 96-well plate to a final concentration of 1μM, and 2 replicates were set. For samples at T=0 minutes, the compound and cells were thoroughly mixed for 1 minute, and then 25μL of sample was immediately added to 125μL of stop solution (containing 200ng / mL tolbutamide and 200ng / mL labetalol in acetonitrile) in an ice bath and mixed. At the same time, all plates were placed in an incubator at 37°C and 5% CO2, with the shaker set to 600rpm. The samples were mixed at 15, 30, 60, and 90 minutes of incubation, and 25μL of sample was added to 125μL of stop solution (containing 200ng / mL tolbutamide and 200ng / mL labetalol in acetonitrile) in an ice bath, and shaken at 500rpm for 10 minutes after mixing. The plates were then centrifuged at 3220 × g for 20 minutes at 4°C. After centrifugation, 80 μL of supernatant was transferred from each well to another 96-well plate containing 240 μL of ultrapure water. The intrinsic clearance (CLint) and half-life (T1 / 2) were then calculated using LC-MS / MS.

[0595] Experimental Example 4 Pharmacokinetic Test in Rats

[0596] In this experiment, the pharmacokinetic behavior of the test compound was investigated in SD rats after intravenous (IV) and oral (PO) administration.

[0597] On the day of administration, the actual body weight of the rats was weighed and the administration volume was calculated. There were 3 rats in each group, and two groups of tests were performed for each compound, one group was administered with a single intravenous injection, and the other group was administered with a single oral gavage. Whole blood samples were collected at the specified time (0.25, 0.5, 1, 2, 4, 8, 24h after administration) by blood sampling from the jugular vein. After blood sample collection, it was immediately transferred to a labeled commercial sample tube containing K2-EDTA (0.85-1.15mg), followed by centrifugation (3200x g, 4°C, 10 minutes) and plasma was collected. The plasma was transferred to a pre-cooled centrifuge tube, quick-frozen in dry ice, and then stored in an ultra-low temperature freezer at -60°C or lower until LC-MS / MS analysis.

[0598] Plasma concentrations were determined using LC-MS / MS. Plasma concentration data were analyzed using WinNonlin Version 6.3 (Pharsight, Mountain View, CA) pharmacokinetic software using a non-compartmental model. Pharmacokinetic parameters were calculated using the linear-log trapezoidal method.

[0599] Experimental Example 5 Pharmacokinetic Test in Mice

[0600] In this experiment, the pharmacokinetic behavior of the test compound was investigated in BALB / c mice after intravenous (IV) and oral (PO) administration.

[0601] On the day of administration, the actual body weight of the mice was weighed and the administration volume was calculated. There were 9 mice in each group, and two groups of tests were performed for each compound, one group was administered with a single intravenous injection, and the other group of mice was administered with a single oral gavage. Whole blood samples were collected at the specified time (0.25, 0.5, 1, 2, 4, 8, and 24 hours after administration) by orbital bleeding. After blood sample collection, it was immediately transferred to a labeled commercial sample tube containing K2-EDTA (0.85-1.15 mg), followed by centrifugation (3200x g, 4°C, 10 minutes) and plasma was collected. The plasma was transferred to a pre-cooled centrifuge tube, quickly frozen in dry ice, and then stored in an ultra-low temperature freezer at -60°C or lower until LC-MS / MS analysis.

[0602] Plasma concentrations were determined using LC-MS / MS. Plasma concentration data were analyzed using WinNonlin Version 6.3 (Pharsight, Mountain View, CA) pharmacokinetic software using a non-compartmental model. Pharmacokinetic parameters were calculated using the linear-log trapezoidal method.

[0603] Comparative compounds A / C / D are compounds 73 / 42 / 125 in patent application WO2022249060, respectively, which were prepared by the synthesis method reported therein. Comparative compound C is HRO761, a compound developed by Novartis and has entered clinical development.

[0604] Comparative compound B was prepared by referring to the synthesis method of compound A20, using A1-4 instead of A20-5. ESI (m / z) = 696.3 [M+H] + .1H NMR (400MHz, DMSO-d6) δ8.35(s,2H),8.08(dt,J=5.4,3.2Hz,2H),8.01(d,J=8.6Hz,1H),7.95(t,J=1.3Hz,1H),7.72–7.66(m,1H),7.54–7.48(m ,3H),5.37(s,2H),4.53(d,J=11.6Hz,1H),3.58–3.22(m,4H),3.00(d,J =7.6Hz, 3H), 2.84 (d, J = 11.0Hz, 1H), 2.38 (s, 3H), 1.19 (t, J = 7.3Hz, 3H).

[0605] Experimental Example 6 hERG inhibition test

[0606] HEK293 cells were cultured in DMEM medium containing 10% fetal bovine serum and 0.8 mg / mL G418 at 37°C and 5% CO2. TM After express digestion, centrifugation was performed and the cell density was adjusted to 2 × 10 6 cells / mL, then gently mix the cells on a room temperature balanced shaker for 15-20 minutes, and perform patch clamp detection on the machine. The culture medium of the prepared cells was replaced with extracellular fluid. The intracellular and extracellular fluids were aspirated from the liquid pool and added to the intracellular fluid pool, cell and test substance pool of the QPlate chip respectively. The whole-cell patch clamp records the voltage stimulation of the whole-cell hERG potassium current, and the experimental data is collected and stored by Qpatch. The compound started at 30μM, diluted 3 times, and 6 concentration points were set. Each drug concentration was set to be administered twice for at least 5 minutes. The current detected in the external fluid without the compound for each cell was used as its own control group, and at least two cells were used for each concentration to repeat the test twice independently. All electrophysiological experiments were performed at room temperature.

[0607] Data analysis: First, the current after each drug concentration is normalized with the blank control current. Then calculate the inhibition rate corresponding to each drug concentration For each concentration, calculate the mean and standard error, and calculate the half-inhibitory concentration for each compound: The above equation was used to perform nonlinear fitting of the dose-dependent effect, where Y represents the inhibition rate, C represents the concentration of the test substance, and IC 50 is the half-inhibitory concentration, and HillSlope represents the Hill coefficient. Curve fitting and IC 50 The calculations were completed using Graphpad software.

[0608] Experimental Example 7 Cytochrome oxidase P450 inhibition test

[0609] 1) Preparation of buffer solution:

[0610] 100 mM K-Buffer: Mix 9.5 mL of stock solution A with 40.5 mL of stock solution B, adjust the total volume to 500 mL with ultrapure water, and titrate the buffer to pH 7.4 with KOH or H3PO4.

[0611] Raw material A (1M potassium dihydrogen phosphate): 136.5 g potassium dihydrogen phosphate in 1 L water;

[0612] Stock B (1M KH2PO4): 174.2 g KH2PO4 in 1 L water.

[0613] 2) Preparation of test substance

[0614] The test substance powder is prepared into a stock solution of a certain concentration using DMSO or other organic solvents, and then further diluted with a suitable organic solvent.

[0615] 3) In vitro incubation

[0616] The in vitro incubation system of liver microsomes for CYP450 enzyme metabolic phenotype studies is a biochemical reaction carried out under conditions simulating physiological temperature and physiological environment, with the prepared liver microsomes supplemented with redox coenzymes and enzyme-specific selective inhibitors.

[0617] 4) Detection of parent drug or metabolites

[0618] The concentration of parent drug or its metabolites in the incubation solution was determined by LC-MS / MS.

[0619] Experimental Example 8 Mouse Tumor Pharmacodynamic Model

[0620] In this experimental example, the in vivo efficacy of the test compound was evaluated in a mouse xenograft tumor model after oral administration (PO).

[0621] Human colon adenocarcinoma cell line SW48 was cultured in monolayer in vitro under the following conditions: DMEM medium containing 10% fetal bovine serum, 1% penicillin and streptomycin, cultured at 37°C with 5% CO2. Human colon adenocarcinoma cell line HCT116 was cultured in monolayer in vitro under the following conditions: McCoy's 5A medium containing 10% fetal bovine serum, 1% penicillin and streptomycin, cultured at 37°C with 5% CO2. The cells were digested and passaged twice a week using trypsin. When the cell saturation reached 80%-90%, the cells were harvested, counted, and inoculated. 0.1 mL (10 7 ) SW48 cells or 0.1 mL (5*10 6 HCT116 cells were subcutaneously inoculated on the right back of each mouse. On day 14 after cell inoculation, the average tumor volume reached 150-200 mm 3 Around 10 days, the animals were randomly divided into groups and given drugs by gavage once daily. Body weight and tumor volume changes were recorded. After a certain number of days of dosing, the experiment was terminated. Tumor volume changes were statistically analyzed. Tumor growth inhibition rate (TGI%) and p-value were calculated using data from the final day (see the table below).

[0622] Tumor volume calculation formula:

[0623] Tumor volume (mm 3 )=1 / 2×(a×b 2 ) (Note: a represents the major diameter, b represents the minor diameter)

[0624] Tumor growth inhibition rate (TGI%) = [1-(average tumor volume of a certain drug group at the end of drug administration - average tumor volume of the drug group at the beginning of drug administration) / (average tumor volume of the blank control group at the end of treatment - average tumor volume of the blank control group at the beginning of treatment)] × 100%

[0625] Remark:

[0626] The ap value was calculated based on the relative tumor volume of each mouse in different groups. The p value was obtained by comparing the blank control group and each drug-treated group using the One-way ANOVA method. Statistical analysis of p>0.05 indicated that there was no significant difference between each group and the blank control group, statistical analysis of p≤0.05 indicated that there was a significant difference between the drug-treated group and the blank control group, and statistical analysis of p≤0.01 indicated that there was a very significant difference between the drug-treated group and the blank control.

[0627] The bp value is the p value obtained by comparing the control compound administration group and the compound administration group using the One-way ANOVA method. The statistical analysis of p>0.05 indicates that there is no significant difference between the compound administration group and the control compound administration group, the statistical analysis of p≤0.05 indicates that there is a significant difference between the compound administration group and the control compound administration group, and the statistical analysis of p≤0.01 indicates that there is a very significant difference between the compound administration group and the control compound administration group.

[0628] c. The experiment of A181 and control compound C was not completed on the same day, so the p-value of the two groups of data on the last day could not be directly calculated. The reported p-value was determined by the tumor volume on day 16 (145±28mm in the A181 group). 3 , compared with compound C group 688±135mm 3 ) calculated.

[0629] In addition to those described herein, various modifications of the present invention will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. Each reference cited in this application (including all patents, patent applications, journal articles, books, and any other disclosures) is incorporated herein by reference in its entirety.

Claims

1. A compound or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, wherein the compound has the structure of formula (I): in: Selected from Where R 1 Connected to the position marked with "*" in the above group, R 2 Connected to the position marked with "**" in the above group, R 3 Connected to the position marked with "***" in the above group, and R 4 Connected to the position marked with "****" in the above group; R 1 Selected from -NR 11 R 12 , R 2 for R 4 for R 3 , R 11 , R 12 , R 13 , R 21 , R 22 , R 23 and R 24 Each occurrence is independently selected from H, halogen, -OH, -NH2, -CN, -NO2, -SF5, C 1-6 Alkyl, deuterated C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl, C 6-12 Aralkyl, -C(=O)R a 、-OC(=O)R a 、-C(=O)OR a 、-OR a 、-SR a 、-S(=O)R a 、-S(=O)2R a 、-S(=O)2NR a R b 、-NR a R b 、-C(=O)NR a R b 、-NR a -C(=O)R b 、-NR a -C(=O)OR b 、-NR a -S(=O)2-R b 、-NR a -C(=O)-NR a R b 、-P(=O)R a R b , -C 1-6 Alkylene-R a , -C 1-6 Alkylene-OR a , -C 1-6 Alkylene-NR a R b 、-OC 1-6 Alkylene-NR a R b ,(-C 3-6 Cycloalkylene)-CN and (-C 3-6 Cycloalkylene)-C 1-6 Haloalkyl; R 14 Each occurrence is independently selected from -L 1 -(C 3-6 Cycloalkyl), -L 1 -(3-10 membered heterocyclic group), -L 1 -(C 6-10 Aryl) and -L 1 -(5-14 membered heteroaryl); R 41 Selected from C 3-6 Hydrocarbon ring, 3-10 membered heterocyclic ring, C 6-10 Aromatic rings and 5-14 membered heteroaromatic rings; Ring C, ring D, ring E, ring X and ring Z are each independently selected from C 3-6 Hydrocarbon ring, 3-10 membered heterocyclic ring, C 6-10 Aromatic ring and 5-14 members heteroaromatic ring; Ring Y is absent or selected from C 3-6 Hydrocarbon ring, 3-10 membered heterocyclic ring, C 6-10 aromatic ring and 5-14 membered heteroaromatic ring; when ring Y is absent, R 24 Nor does it exist; L 1 is selected from a direct bond, -O-, -C(=O)-, -NRC(=O)-, -S-, -S(=O)-, -S(=O)2-, C 1-6 Alkylene and -O-(C 1-6 Alkylene)-; L 2 Selected from -O-, -C(=O)-, -NRC(=O)-, -S-, -S(=O)-, -S(=O)2-, C 1-6 Alkylene and -O-(C 1-6 Alkylene)-; R, R a and R b Each occurrence is independently selected from H, C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl and C 6-12 Aralkyl; The above alkylene, alkyl, alkenyl, alkynyl, cycloalkylene, cycloalkyl, hydrocarbon ring, heterocyclic radical, heterocyclic ring, aryl, aromatic ring, heteroaryl, heteroaromatic ring and aralkyl are each optionally substituted with one or more substituents independently selected from the following: halogen, -OH, =O, =CF2, -NH2, -CN, -NO2, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl, C 6-12 Aralkyl, -C(=O)R c 、-OC(=O)R c 、-C(=O)OR c 、-OR c 、-SR c 、-S(=O)R c 、-S(=O)2R c 、-S(=O)2NR c R d 、-NR c R d 、-C(=O)NR c R d 、-NR c -C(=O)R d 、-NR c -C(=O)OR d 、-NR c -S(=O)2-R d 、-NR c -C(=O)-NR c R d , -C 1-6 Alkylene-OR c , -C 1-6 Alkylene-NR c R d and-OC 1-6 Alkylene-NR c R d The alkylene, alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and aralkyl groups are further optionally substituted by one or more substituents independently selected from the following: halogen, -OH, =O, -C(=O)O-tert-butyl, -NH2, -CN, -NO2, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl, C 6-12 Aralkyl, -OC 1-6 Alkyl and -C 1-6 Alkylene-OC 1-6 alkyl; R c and R d Each occurrence is independently selected from H, C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl and C 6-12 Aralkyl, the alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and aralkyl are further optionally substituted by one or more substituents independently selected from the following: halogen, -OH, =O, -C(=O)O-tert-butyl, -NH2, -CN, -NO2, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl, C 6- 12 Aralkyl and -C 1-6 Alkylene-OC 1-6 alkyl; n, p and q are each independently an integer selected from 1, 2 or 3; and m is an integer of 0 or 1.

2. The compound of claim 1 or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein the compound has the structure of the following formula:

3. The compound of claim 1 or 2, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, wherein: R 1 for R 13 is H, halogen or C 1-6 alkyl; R 14 is selected from 3-10 membered heterocyclic group, -(C 1-6 Alkylene)-(3-10 membered heterocyclic group) and -O-(C 1-6 alkylene)-(3-10 membered heterocyclyl), said heterocyclyl being optionally substituted by one or more substituents independently selected from the following: C 1-6 Alkyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, -C 1-6 Alkylene-C 3-6 Cycloalkyl and -C 1-6 Alkylene-CN; and n is an integer of 1 or 2; Preferably, R 1 for 4. The compound of claim 1 or 2, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, wherein: R 1 for Ring C is a benzene ring, a 5-6 membered heterocyclic ring or a 5-6 membered heteroaromatic ring; Ring D is a benzene ring or a 5-6 membered heteroaromatic ring; The above-mentioned benzene ring, heterocyclic ring and heteroaromatic ring are each optionally substituted by one or more substituents independently selected from the following: halogen, -OH, -NH2, -CN, -NO2, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Haloalkyl, -OC 1-6 Alkyl, C 3-6 Preferably, the above-mentioned benzene ring, heterocyclic ring and heteroaromatic ring are each optionally substituted by one or more substituents independently selected from the following: halogen, C 1-6 Alkyl, C 1-6 Haloalkyl, -OC 1-6 Alkyl and C 3-6 Cycloalkyl; Preferably, R 1 for 5. The compound of claim 1 or 2, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, wherein R 1 For-NR 11 R 12 ; R 11 and R 12 Each independently selected from H, C 1-6 Alkyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl, C 6-12 Aralkyl, -C 1-6 Alkylene-R a and -C 1-6 Alkylene-NR a R b ; R a and R b Each occurrence is independently selected from H, C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl and C 6-12 Aralkyl; The alkylene, alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and aralkyl groups are each optionally substituted at each occurrence with one or more substituents independently selected from the group consisting of halogen and C 1-6 alkyl; Preferably, R 1 for 6. The compound of claim 1 or 2, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, wherein: R 1 for Ring E is C 3-6 Hydrocarbon ring, 3-10 membered heterocyclic ring, C 6-10 Aromatic ring or 5-14 membered heteroaromatic ring, the hydrocarbon ring and heterocyclic ring are optionally substituted by =CF2; R 13 H, halogen, C 1-6 Alkyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic, -C(=O)-(C 1-6 Alkyl), -O-(C 1-6 Alkyl) or -N(C 1-6 alkyl)2, wherein the alkyl, cycloalkyl and heterocyclic groups are further optionally C 1-6 Alkyl, C 3-6 Cycloalkyl or 3-10 membered heterocyclic group substitution; n is an integer of 1 or 2; Preferably, R 1 for 7. The compound of any one of claims 1 to 6, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein when Y is absent, then a) Ring X is a 5-6 membered heterocyclic ring (eg, a pyridone ring) or a 5 membered heteroaromatic ring; or b) Ring X is a benzene ring, and at least one R 23 (-C 3-6 Cycloalkylene)-CN or (-C 3-6 Cycloalkylene)-C 1-6 Halogenated alkyl.

8. The compound of any one of claims 1 to 6, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, wherein ring X is a benzene ring and ring Y is a 5-6 membered heterocyclic ring or a 5-6 membered heteroaromatic ring.

9. The compound of any one of claims 1 to 8, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound, or prodrug thereof, wherein R 21 , R 22 , R 23 and R 24 Each occurrence is independently selected from H, halogen, -SF5, C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 3-6 Cycloalkyl, -S(=O)2-(C 1-6 alkyl), -S(=O)2-(C 3-6 Cycloalkyl), -P(=O)(C 1-6 Alkyl)2, (-C 3-6 Cycloalkylene)-CN and (-C 3-6 Cycloalkylene)-C 1-6 Halogenated alkyl.

10. The compound of any one of claims 1 to 9, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound, or prodrug thereof, wherein R 2 Selected from:

11. The compound of any one of claims 1 to 6 or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein X is a benzene ring or a 5-6 membered heteroaromatic ring (e.g., a pyridine ring); and Y is absent; R 23 Each occurrence is independently selected from H, halogen, -SF5, C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 3-6 Cycloalkyl, -S(=O)2-(C 1-6 alkyl), -S(=O)2-(C 3-6 Cycloalkyl), -P(=O)(C 1-6 Alkyl)2, (-C 3-6 Cycloalkylene)-CN and (-C 3-6 Cycloalkylene)-C 1-6 Haloalkyl; Preferably, R 2 Selected from:

12. The compound of any one of claims 1 to 11, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound, or prodrug thereof, wherein R 3 H or C 1-6 Alkyl; preferably ethyl.

13. The compound of any one of claims 1 to 12, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, wherein ring Z is a 3-10 membered heterocycle or a benzene ring; preferably a 5-10 membered heterocycle; more preferably a 5-6 membered heterocycle; and The heterocyclic ring and the benzene ring are each optionally substituted at each occurrence by one or more substituents independently selected from the group consisting of halogen and C 1-6 alkyl; Preferably, ring Z is 14. The compound of any one of claims 1 to 13, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, wherein L 2 is -C(=O)- or -NRC(=O)-, wherein R is H or C 1-6 alkyl; Preferably, L 2 It is -C(=O)-.

15. The compound of any one of claims 1 to 14, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound, or prodrug thereof, wherein R 41 Selected from 3-10 membered heterocyclic ring, C 6-10 aromatic ring and 5-14 membered heteroaromatic ring, wherein the heterocyclic ring, aromatic ring and heteroaromatic ring are each optionally substituted by one or more substituents independently selected from the following: halogen, -OH, -CN, C 1-6 Alkyl, -OC 1-6 Alkyl and -SC 1-6 Alkyl, preferably, the heterocyclic ring, aromatic ring and heteroaromatic ring are at least -OH, -CN or -OC 1-6 Alkyl substitution; Preferably, -L 2 -R 41 for 16. The compound of any one of claims 1 to 15, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound, or prodrug thereof, wherein R 41 is a 5-6 membered heteroaromatic ring (preferably a pyrazole ring or a pyrimidine ring), which is substituted by at least one -OH; Preferably, R 41 is a 5-membered heteroaromatic ring (preferably a pyrazole ring) substituted by at least one -OH; Preferably, -L 2 -R 41 for Preferably, -L 2 -R 41 for 17. A compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein the compound is selected from:

18. The compound of any one of claims 1 to 17, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound, or prodrug thereof, wherein the compound is in a non-zwitterionic form, a zwitterionic form, or a mixture of zwitterionic and non-zwitterionic forms; Preferably, the compound is in any one of the following forms or a mixture of any two or three of them:

19. A pharmaceutical composition comprising a preventively or therapeutically effective amount of a compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, and a pharmaceutically acceptable carrier.

20. Use of a compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, or a pharmaceutical composition according to claim 19 in the preparation of a medicament for use as a WRN inhibitor, preferably, the medicament is used to prevent or treat cancer (preferably, the cancer is characterized by microsatellite high instability (MSI-H) or mismatch repair deficiency (dMMR)); preferably, the cancer is selected from colorectal cancer, gastric cancer, endometrial cancer, uterine cancer, adrenocortical carcinoma, cervical cancer, esophageal cancer, breast cancer, kidney cancer, prostate cancer and ovarian cancer.

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