Fused ring compound, pharmaceutical composition containing same and use thereof

Abstract

The present invention relates to a compound of formula (I), a pharmaceutical composition containing same, and the use thereof for preventing or treating a disease.

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.2020Oct; 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] represents a single bond or a double bond, provided that the two double bonds are not directly connected;

[0010] W 1 、W 2 、W 3 and W 4 are each independently C or N, provided that C is connected to a double bond; preferably, W 1 and W 2 At least one of them is N, and / or W 3 and W 4 At least one of is N;

[0011] Selected from

[0012] R 1 、R 3 、R 21 and R 22 Each occurrence is independently selected from H, deuterium atoms, halogen, -OH, -NH2, -CN, -NO2, -SF5, =CH2, 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 、-S(=O)(=NR 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;

[0013] When m is greater than 1, two R 3 Together with the group to which it is attached, it optionally forms C 3-6 hydrocarbon ring, 3-10 membered heterocyclic ring, C 6-10 Aromatic ring or 5-14 membered heteroaromatic ring;

[0014] R 4 for

[0015] 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)-;

[0016] 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;

[0017] 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;

[0018] Ring B, 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;

[0019] Ring Y is absent or selected from C 3-6 hydrocarbon ring, 3-10 membered heterocyclic ring, C 6-10 aromatic rings and 5-14 membered heteroaromatic rings; when ring Y is absent, R 22 It does not exist either;

[0020] The above alkylene, alkyl, alkenyl, alkynyl, cycloalkylene, cycloalkyl, hydrocarbon ring, heterocyclic group, heterocycle, aryl, aromatic ring, heteroaryl, heteroaryl ring and aralkyl groups are each optionally substituted with one or more substituents independently selected from the group consisting of deuterium atoms, halogen, -OH, =O, -NH2, -CN, -NO2, =CH2, C 1-6 Alkyl, deuterated 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 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 , wherein each of the alkylene, alkyl, alkenyl, =CH2, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and aralkyl groups is further optionally substituted by one or more substituents independently selected from the group consisting of 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, -C 1-6 Alkylene-C 3-6 Cycloalkyl, -OC 1-6 Alkyl and -C 1-6 Alkylene-OC 1-6 alkyl;

[0021] 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; and

[0022] p, q and m are each independently an integer selected from 1, 2 or 3.

[0023] 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.

[0024] 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.

[0025] 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.

[0026] 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.

[0027] Detailed Description of the Invention

[0028] definition

[0029] 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.

[0030] 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.

[0031] 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.

[0032] 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).

[0033] 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.

[0034] 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", "C2-6 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.

[0035] 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.

[0036] 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.

[0037] 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.

[0038] 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.

[0039] 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.

[0040] 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.

[0041] 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,

[0042] As used herein, the term "heterocyclyl" (or "heterocycle") encompasses bridged heterocyclyls (bridged heterocycle) and spiro heterocyclyls (spiro heterocycle).

[0043] 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.

[0044] 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.

[0045] 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.

[0046] 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.

[0047] 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.

[0048] As used herein, the term "halo" or "halogen" group is defined to include F, Cl, Br, or I.

[0049] 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.

[0050] 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.

[0051] 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.

[0052] 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.

[0053] 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.

[0054] As used herein, the term "one or more" means 1 or more than 1, such as 2, 3, 4, 5 or 10, where reasonable.

[0055] Unless otherwise indicated, as used herein, the point of attachment of a substituent may be from any suitable position of the substituent.

[0056] 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.

[0057] 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 15N); 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.

[0058] 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%).

[0059] 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).

[0060] Atropisomers are compounds that can be separated into rotationally restricted isomers.

[0061] 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.

[0062] Pharmaceutically acceptable salts of the compounds of the present invention include acid addition salts and base addition salts thereof.

[0063] 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.

[0064] 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.

[0065] 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.

[0066] 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.

[0067] 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)).

[0068] 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.

[0069] As used herein, the term "about" means within ±10%, preferably within ±5%, and more preferably within ±2% of the stated numerical value.

[0070] Compound

[0071] 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):

[0072] in:

[0073] represents a single bond or a double bond, provided that the two double bonds are not directly connected;

[0074] W 1 、W 2 、W 3 and W 4 are each independently C or N, provided that C is connected to a double bond; preferably, W 1 and W 2 At least one of them is N, and / or W 3 and W 4 At least one of is N;

[0075] Selected from

[0076] R 1 、R 3 、R 21 and R 22Each occurrence is independently selected from H, deuterium atoms, halogen, -OH, -NH2, -CN, -NO2, -SF5, =CH2, 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 、-S(=O)(=NR 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;

[0077] When m is greater than 1, two R 3Together with the group to which it is attached, it optionally forms C 3-6 hydrocarbon ring, 3-10 membered heterocyclic ring, C 6-10 Aromatic ring or 5-14 membered heteroaromatic ring;

[0078] R 4 for

[0079] 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)-;

[0080] 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;

[0081] 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;

[0082] Ring B, 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;

[0083] 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 22 It does not exist either;

[0084] The above alkylene, alkyl, alkenyl, alkynyl, cycloalkylene, cycloalkyl, hydrocarbon ring, heterocyclic group, heterocycle, aryl, aromatic ring, heteroaryl, heteroaryl ring and aralkyl groups are each optionally substituted with one or more substituents independently selected from the group consisting of deuterium atoms, halogen, -OH, =O, -NH2, -CN, -NO2, =CH2, C 1-6 Alkyl, deuterated 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)Rc 、-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 , wherein each of the alkylene, alkyl, alkenyl, =CH2, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and aralkyl groups is further optionally substituted by one or more substituents independently selected from the group consisting of 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, -C 1-6 Alkylene-C 3-6 Cycloalkyl, -OC 1-6 Alkyl and -C 1-6 Alkylene-OC 1-6 alkyl;

[0085] 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 6-12Aralkyl, 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; and

[0086] p, q and m are each independently an integer selected from 1, 2 or 3.

[0087] 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:

[0088] In some embodiments, Ring B is C 3-6 a hydrocarbon ring or a 3-10 membered heterocyclic ring.

[0089] In a preferred embodiment, ring B is a cyclopentene ring, a cyclohexene ring, a pyrrolidine ring, an oxazolidine ring, a piperidine ring, a morpholine ring or an azepane ring.

[0090] In some embodiments, R 1 Each occurrence is independently selected from H, halogen, C 1-6 Alkyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl, -S(=O)2R a 、-OR a and -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; preferably, R 1 Each occurrence is independently selected from 3-10 membered heterocyclyl, C 6-10 Aryl, 5-14 membered heteroaryl and -NR aR b wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl groups are each optionally substituted by one or more substituents independently selected from the following: halogen, -S(=O)2R c 、C 1-6 Alkyl, C 2-6 Alkenyl, =CH2, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl and 5-14 membered heteroaryl; said alkyl, alkenyl, =CH2, cycloalkyl, heterocyclyl, aryl and heteroaryl are each further optionally substituted by one or more substituents independently selected from the following: halogen, C 1-6 Alkyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group and -C 1-6 Alkylene-C 3- 6-cyclic hydrocarbon group.

[0091] In some embodiments, R 1 Each occurrence is independently selected from H, halogen, C 1-6 Alkyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl, 5-14 membered heteroaryl, -OR a and -NR a R b , preferably, R 1 Each occurrence is independently selected from 3-10 membered heterocyclyl, C 6-10 Aryl, 5-14 membered heteroaryl and -NR a R b wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each optionally substituted by one or more substituents independently selected from the following: halogen, C 1-6 Alkyl, C 2-6 Alkenyl, =CH2, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl and 5-14 membered heteroaryl; said alkyl, alkenyl, =CH2, cycloalkyl, heterocyclyl, aryl and heteroaryl are each further optionally substituted by one or more substituents independently selected from the following: halogen, C 1-6 Alkyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group and -C 1-6 Alkylene-C 3-6 Cyclic hydrocarbon group.

[0092] In some embodiments, R 1 Each occurrence is independently C 6-10 Aryl, -NR a R b .

[0093] In some embodiments, R 1 Each occurrence is independently -NR a R b .

[0094] In a preferred embodiment, R 1 is H, methyl, halogen, methoxy,

[0095] In a preferred embodiment, R 1 is H, methyl, halogen, methoxy, More preferably

[0096] In a preferred embodiment, R 1 is H, methyl, halogen, methoxy, More preferably

[0097] In some embodiments, R 1 for

[0098] In some embodiments, Ring X is a benzene ring, a 5-6 membered heterocyclic ring, or a 5-6 membered heteroaryl ring and Ring Y is absent.

[0099] In some embodiments, Ring X is a benzene ring and Ring Y is C 3-6 hydrocarbon ring, benzene ring, 5-6 membered heterocyclic ring or 5-6 membered heteroaromatic ring.

[0100] In some embodiments, for More preferably

[0101] In some embodiments, for More preferably

[0102] In some embodiments, R 21 and R 22 Each occurrence is independently selected from H, halogen, -SF5, C 1-6 Alkyl, C 3-6Cycloalkyl, 3-10 membered heterocyclic group, -O-(C 1-6 alkyl), -S(=O)2-(C 1-6 alkyl), -S(=O)2-(C 3-6 Cycloalkyl), -S(=O)(=NR a )R b 、-P(=O)(C 1-6 Alkyl)2, (-C 3-6 Cycloalkylene)-CN and (-C 3-6 Cycloalkylene)-C 1-6 Alkyl, said alkyl, cycloalkylene, cycloalkyl and heterocyclyl are each optionally substituted with one or more substituents independently selected from the group consisting of halogen, C 1-6 Alkyl and halogenated C 1-6 alkyl.

[0103] In a preferred embodiment, R 21 and R 22 Each occurrence is independently selected from H, halogen, -SF5, C 1-6 Alkyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group and -O-(C 1-6 alkyl), said alkyl, cycloalkyl and heterocyclyl being each optionally substituted by one or more substituents independently selected from the group consisting of halogen, C 1-6 Alkyl and halogenated C 1-6 alkyl.

[0104] In some embodiments, Selected from:

[0105] In some embodiments, Selected from:

[0106] In some embodiments, R 3 H, C 1-6 Alkyl, -OR a or -SR a Preferably, R 3 H or C 1-6 alkyl.

[0107] In a preferred embodiment, R 3 is H, methyl, ethyl, -O-CH3 or -S-CH3; most preferably, R 3 is H or methyl.

[0108] In a preferred embodiment, when m is greater than 1, two R 3 Together with the group to which it is attached, it optionally forms C 3-6 A hydrocarbon ring (preferably a cyclopropyl ring) optionally substituted by one or more substituents independently selected from the group consisting of halogen, C 1-6 Alkyl and halogenated C 1-6 alkyl.

[0109] In some embodiments, 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

[0110] 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, C 1-6 Alkyl and halogenated C 1-6 alkyl.

[0111] In some embodiments, ring Z is

[0112] In some embodiments, L 2 -C(=O)-, C 1-6 Alkylene or -NRC(=O)-, where R is H or C 1-6 alkyl.

[0113] In some embodiments, L 2 -C(=O)- or -NRC(=O)-, wherein R is H or C 1-6 alkyl.

[0114] In a preferred embodiment, L 2 It is -C(=O)-, -CH2-, and -CD2-.

[0115] In a preferred embodiment, L 2 It is -C(=O)-.

[0116] In some embodiments, 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.

[0117] In some embodiments, -L2 -R 41 for

[0118] In some embodiments, -L 2 -R 41 for

[0119] In some embodiments, -L 2 -R 41 for

[0120] In some embodiments, R 41 It is a 5-6 membered heteroaromatic ring, preferably a 6 membered heteroaromatic ring, more preferably a pyridine ring or a pyrimidine ring, which is substituted by at least one -OH group.

[0121] In some embodiments, -L 2 -R 41 for

[0122] The present disclosure encompasses technical solutions / compounds obtained by any combination of the various embodiments.

[0123] 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:

[0124] Pharmaceutical compositions and methods of treatment

[0125] 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.

[0126] 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.

[0127] 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.

[0128] 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.

[0129] 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.

[0130] 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.

[0131] As used herein, unless otherwise indicated, the terms "treat," ...

[0132] 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.).

[0133] In another embodiment, the pharmaceutical compositions of the present invention may further comprise one or more additional therapeutic or prophylactic agents.

[0134] General synthetic route:

[0135] Route 1

[0136] Route 2

[0137] Wherein, in Scheme 1 and Scheme 2, PG is a protecting group, m is 1 or 2, and the remaining groups are as defined herein. Example

[0138] 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.

[0139] The abbreviations used in this invention have the following meanings:

[0140] Example 1: (C87-P1-A, C87-P1-B, C87-P2-A, C87-P2-B)

[0141] 1) Synthesis of intermediate C87-2:

[0142] Compound C87-1 (18.46 g, 128.2 mmol) was added to tetrahydrofuran (600 mL), cooled to 0°C, and sodium hydride (5.12 g, 128.2 mmol) was slowly added. The atmosphere was replaced with nitrogen three times, and the reaction was stirred at 0°C for half an hour. Then, n-butyllithium solution (80 mL, 128.2 mmol) was slowly added dropwise, and the reaction was stirred at 0°C under nitrogen for half an hour. Ethyl propionyl acetate (25 g, 128.2 mmol) was then added to the reaction solution. After stirring at 0°C for half an hour, the mixture was slowly warmed to room temperature and allowed to react overnight. TLC indicated that the reaction of the starting material was substantially complete. The reaction solution was quenched with saturated ammonium chloride solution and extracted with ethyl acetate. The organic phase was then washed with saturated brine and dried over anhydrous sodium sulfate. The filtrate was filtered and concentrated under reduced pressure. The residue was purified by column chromatography (PE / EA = 90 / 10) to afford C87-2 (11.96 g, 36.2% yield) as a yellow oil.1 H NMR (400MHz, CDCl3): δ4.21-4.10(m,2H),4.05-3.98(m,2H),3.77(s,2H),3.71-3.65(m,1H),2.83-2.76(m,1H) ,1.94-1.85(m,1H),1.60-1.51(m,1H),1.35(s,3H),1.28(m,3H),1.23(t,J=9.6Hz,3H),1.11(d,J=9.6Hz,3H).

[0143] 2) Synthesis of intermediate C87-4:

[0144] Compound C87-2 (11.96 g, 46.3 mmol), C87-3 (7.4 g, 46.3 mmol), and anhydrous p-toluenesulfonic acid (0.80 g, 4.63 mmol) were added to n-butanol (15 mL). The reaction solution was stirred at 140°C under nitrogen for 16 hours. LC-MS showed that the starting materials were completely reacted. The reaction solution was cooled and concentrated under reduced pressure. The residue was purified by column chromatography (DCM / MeOH = 85 / 15) to obtain a white solid C87-4 (10.1 g, yield 69.6%). LCMS (ESI) m / z: 315.0 [M+H] + .

[0145] 3) Synthesis of intermediate C87-5:

[0146] Compound C87-4 (10.1 g, 32.1 mmol) and triphenylphosphine (12.6 g, 48.2 mmol) were added to tetrahydrofuran (300 mL) and cooled to 0°C. DIAD (9.57 g, 48.2 mmol) was then slowly added dropwise. The mixture was slowly warmed to room temperature under a nitrogen atmosphere and stirred overnight. LC-MS indicated complete reaction of the starting materials. The reaction solution was cooled and concentrated under reduced pressure. The residue was purified by column chromatography (DCM / MeOH = 85 / 15) to afford C87-5 (7.46 g, 78.3% yield) as a white solid. 1 H NMR (400MHz, DMSO-d6): δ8.14-8.12(m,2H),7.55-7.52(m,3H),5.95-5.95(m,1H),5.03-5.98(m,1H),4.76-4.74(m,1 H),4.49-4.10(m,1H),3.72-3.67(m,1H),3.54-3.36(m,1H),2.62-2.45(m,1H),2.11-1.90(m,1H),1.39-1.31(m,3H).

[0147] 4) Synthesis of intermediate C87-6:

[0148] TEMPO (0.83 g, 5.3 mmol) was added to an aqueous solution of NaH2PO4 (200 mL, 133.1 mmol, 0.67 M), followed by a suspension of compound C87-5 (7.88 g, 26.6 mmol) in acetonitrile (50 mL). Finally, a solution of NaClO2 (4.79 g, 53.2 mmol) and NaClO solution (0.8 mL) in water (40 mL) was added. The atmosphere was purged with nitrogen three times, then the temperature was raised to 50°C and the reaction was stirred under a nitrogen atmosphere overnight. LC-MS showed that the starting material had reacted completely. The reaction solution was cooled to room temperature and purified by reverse phase column chromatography (H2O / ACN = 85 / 15) to afford C87-6 (5.0 g, 61.1% yield) as a white solid. 1 H NMR (400MHz, DMSO-d6): δ8.15-8.12(m,2H),7.58-7.53(m,3H),7.31-7.05(m,2H),5.97 (s,1H),4.84-4.80(m,1H),2.83-2.73(m,1H),2.02-1.97(m,1H),1.34(d,J=9.6Hz,3H).

[0149] 5) Synthesis of intermediate C87-8:

[0150] Compound C87-6 (500 mg, 1.61 mmol) and two drops of DMF were added to DCE (15 mL), and then POCl3 (500 mg, 3.22 mmol) was slowly added dropwise. The mixture was replaced with nitrogen three times, and the temperature was raised to 80°C. The reaction mixture was stirred for 1 hour under a nitrogen atmosphere. LC-MS showed that the reaction of the raw material was complete. The reaction solution was cooled to room temperature, and a pyridine (2 mL) solution of compound C87-7 (1.58 g, 8.06 mmol) was slowly added. The temperature was raised to 80°C. The reaction mixture was stirred for 1 hour under a nitrogen atmosphere. LC-MS showed that the reaction of the raw material was complete. The reaction solution was cooled to room temperature, washed with saturated aqueous sodium bicarbonate solution, extracted with ethyl acetate, and the organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. It was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (PE / EA=50 / 50) to obtain a yellow solid C87-8 (340 mg, yield 43.3%). 1H NMR (400MHz, DMSO-d6): δ10.60(s,1H),8.23-8.10(m,2H),8.04-8.02(m,2H),7.77-7.75(m,1H),7.61-7.54(m ,4H),5.70(d,J=10.8Hz,1H),3.56-3.45(m,1H),2.78-2.71(m,1H),2.48-2.41(m,1H),1.41(d,J=9.2Hz,3H).

[0151] 6) Synthesis of intermediate C87-9:

[0152] Compound C87-8 (340 mg, 0.70 mmol) was added to chloroform (15 mL), and NBS (248 mg, 1.39 mmol) was slowly added at room temperature. The atmosphere was replaced with nitrogen three times, and the reaction solution was heated to 50°C and stirred under a nitrogen atmosphere overnight. LC-MS showed that the starting material was completely reacted. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The residue was purified by column chromatography (PE / EA = 65 / 35) to obtain a yellow solid C87-9 (360 mg, yield 74.7%). LCMS (ESI) m / z: 566.0 [M+H] + .

[0153] 7) Synthesis of intermediate C87-10:

[0154] Compound C87-9 (218 mg, 0.38 mmol), piperazine (664 mg, 7.7 mmol), and AgBF4 (150 mg, 0.77 mmol) were added to DMSO (5 mL). The atmosphere was replaced with nitrogen three times. The reaction solution was heated to 120°C and stirred under a nitrogen atmosphere for 2 hours. LC-MS showed that the starting materials were completely reacted. The reaction solution was cooled to room temperature, washed with water, and extracted with ethyl acetate. The organic phase was washed with saturated brine and dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (DCM / MeOH = 85 / 15, 0.5% NH3.H2O) to obtain a yellow solid C87-10 (123 mg, yield 55.9%). LCMS (ESI) m / z: 572.2 [M+H] + .

[0155] 8) Synthesis of C87-P1 and C87-P2:

[0156] Compound C87-11 (24 mg, 0.15 mmol) was added to DMF (2 mL), and HOBT (17 mg, 0.12 mmol) and EDCI (32 mg, 0.16 mmol) were added at room temperature. The mixture was stirred at room temperature for 2 hours under nitrogen protection. Then, compound C87-10 (63 mg, 0.11 mmol) and DIEA (43 mg, 0.32 mmol) were added. The nitrogen atmosphere was replaced three times and stirred at room temperature for 2 hours under nitrogen protection. LC-MS showed that the raw material reaction was complete. The reaction solution was concentrated under reduced pressure and the residue was purified by preparative high performance liquid chromatography (RP-PREP-3 SunFire C18 5um 19*150mm 18min-55-65B, A:H2O (0.2% FA), B: ACN, UV: 214 nm, flow rate 15 ml / min) to give C87-P1 (8.0 mg, yield 10.2%, retention time 10.79 minutes) and C87-P2 (10.2 mg, yield 13.0%, retention time 11.59 minutes).

[0157] C87-P1 1 H NMR (400MHz, DMSO-d6): δ10.56(s,1H),8.55(s,1H),8.08-8.06(m,2H),7.98-7.96(m,2H),7.74-7.72(m,1H),7.51-7.50(m,3H),5.71-5. 67(m,1H),3.78-3.73(m,1H),3.38-3.10(m,9H),2.65-2.62(m,1H),2.44-2.41(m,4H),1.52(d,J=7.2Hz,3H).LCMS(ESI)m / z:707.9[M+H] + .

[0158] C87-P2 1 H NMR (400MHz, DMSO-d6): δ10.56(s,1H),8.55(s,1H),8.10-8.07(m,2H),7 .98-7.94(m,2H),7.75-7.73(m,1H),7.52-7.50(m,3H),5.62-5.59(m,1H ),3.77-3.73(m,1H),3.34-3.10(m,9H),3.07-2.99(m,1H),2.44(s,3H), 2.15(d,J=13.6Hz,1H),1.44(d,J=7.2Hz,3H).LCMS(ESI)m / z:707.9[M+H] + .

[0159] 9) Synthesis of C87-P1-A, C87-P1-B, C87-P2-A, and C87-P2-B:

[0160] Compound C87-P1 (8.0 mg) was subjected to chiral separation (IBN, ACN:IPA:TFA=90:10:0.3, 25 ml / min, 254 nm) to give C87-P1-A (2.8 mg, yield 35.0%, retention time 6.98 minutes) and C87-P1-B (2.6 mg, yield 32.5%, retention time 12.46 minutes).

[0161] C87-P1-A 1 H NMR (400MHz, CD3OD): δ8.75(s,1H),8.15(s,2H),8.08(d,J=7.6Hz,1H),7.82(s,1H),7.60(d,J=7.2Hz,1H),7.46(s,3H),5.70(s,1H),3 .87(s,1H),3.87-3.31(m,4H),2.78(s,1H),2.78-2.61(m,3H),2.52(s,1H),1.99-1.65(m,4H),1.28(s,3H).LCMS(ESI)m / z:707.4[M+H] + .

[0162] C87-P1-B 1 H NMR (400MHz, CD3OD): δ8.74(s,1H),8.14(d,J=7.2Hz,2H),8.08(d,J=8.4Hz ,1H),7.82(s,1H),7.60(d,J=8.0Hz,1H),7.46-7.44(m,3H),5.71-5.69(m, 1H),3.86(s,1H),3.85-3.31(m,4H),2.77(s,1H),2.76-2.60(m,3H),2.52( s,1H),1.99-1.65(m,4H),1.29(d,J=8.4Hz,3H).LCMS(ESI)m / z:707.4[M+H] + .

[0163] Compound C87-P2 (10.2 mg) was subjected to chiral separation (IBN, ACN:IPA:TFA=70:30:0.3, 25 ml / min, 254 nm) to give C87-P2-A (3.4 mg, yield 33.3%, retention time 6.02 minutes) and C87-P2-B (3.2 mg, yield 31.3%, retention time 12.52 minutes).

[0164] C87-P2-A 1 H NMR (400MHz, CD3OD): δ8.57(s,1H),8.17-8.16(m,2H),8.08(d,J=8.4Hz,1 H),7.82(s,1H),7.62(d,J=8.4Hz,1H),7.47-7.46(m,3H),5.63(d,J=8.8Hz ,1H),3.87-3.84(m,1H),3.51-3.30(m,8H),3.13-3.08(m,1H),2.52(s,3H) ,2.32(d,J=13.6Hz,1H),1.55(d,J=7.2Hz,3H).LCMS(ESI)m / z:707.5[M+H] + .

[0165] C87-P2-B 1 H NMR (400MHz, CD3OD): δ8.40 (s, 1H), 8.08-8.06 (m, 2H), 7.98 (d, J = 8.4Hz, 1H),7.73-7.72(m,1H),7.54-7.52(m,1H),7.37-7.35(m,3H),5.55-5.51( m,1H),3.78-3.74(m,1H),3.38-3.20(m,8H),3.06-2.98(m,1H),2.41(s, 3H),2.24-2.20(m,1H),1.46(d,J=7.6Hz,3H).LCMS(ESI)m / z:707.5[M+H] + .

[0166] Example 2: (C88-P1, C88-P2, C88-P3)

[0167] 1) Synthesis of intermediate C88-2:

[0168] Compound C87-9 (35 mg, 0.062 mmol), compound C88-1 (104.07 mg, 0.93 mmol), and silver tetrafluoroborate (24.18 mg, 0.124 mmol) were added to dimethyl sulfoxide (4 mL). The reaction solution was heated to 120° C. and reacted for 2 hours. LC-MS showed that the starting materials were completely reacted. Aqueous solution was added dropwise to quench the reaction. The mixture was extracted with ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (DCM / MeOH = 10 / 1) to obtain compound C88-2 (15 mg, 40% yield) as an off-white solid. LCMS (ESI) m / z = 598.1 [M+H] + .

[0169] 2) Synthesis of C88-P1, C88-P2, and C88-P3:

[0170] Compound C87-11 (8.78 mg, 0.06 mmol) was added to DMF (2 mL), and HOBT (5.94 mg, 0.044 mmol) and EDCI (11.52 mg, 0.06 mmol) were added at room temperature. The mixture was stirred at room temperature for 2 hours under nitrogen protection. Compound C88-2 (25 mg, 0.04 mmol) and DIEA (15.48 mg, 0.12 mmol) were then added to the reaction solution, and stirring was continued at room temperature for another 2 hours. LC-MS showed that the reaction of the raw material was complete. The reaction solution was concentrated under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (Waters-PREP-8 SunFire C18 5um 19*150mm 18min-60-75B, A:H2O (0.2% FA), B: ACN, UV: 214 nm, flow rate 15 ml / min) to give C88-P1 (1.3 mg, yield 4.2%, retention time 9.57 minutes), C88-P2 (3.0 mg, yield 9.8%, retention time 10.65 minutes) and C88-P3 (4.2 mg, yield 13.7%, retention time 11.27 minutes). C88-P1 and C88-P2 are both single-configuration compounds, while C88-P3 is a mixture of two diastereomers.

[0171] C88-P1 (the configuration was confirmed by crystal structure, C88-P1 is compound C10): LCMS (ESI) m / z = 734.2 [M+H] + .1HNMR (400MHz, CD3OD): δ8.51(s,1H),8.17-8.14(m,2H),8.10(d,J=8.4Hz,1H),7.83(s,1H),7.62(d,J=9.2Hz,1H),7.47-7.45(m, 3H),5.69-5.66(m,1H),4.58-4.56(m,1H),3.86-3.51(m,6H),2.79-2.76(m,1H),2.50(s,3H),2.18-2.17(m,1H),1.62-1.59(m,7H).

[0172] C88-P2:LCMS(ESI)m / z=734.2[M+H] + . 1H NMR (400MHz, CD3OD): δ8.49(s,1H),8.17-8.15(m,2H),8.06(d,J=8.8Hz,1H),7.82(s,1H),7.62(d,J=8.8Hz,1H),7.47-7.43(m,3H),5.59-5 .56(m,1H),4.74-4.60(m,1H),3.96-3.59(m,6H),3.13-3.05(m,1H), 2.49(s,3H),2.34-2.30(m,1H),2.21-2.17(m,1H),1.56-1.53(m,6H).

[0173] C88-P3:LCMS(ESI)m / z=734.1[M+H] + . 1 H NMR (400MHz, CD3OD): δ8.53(s,1H),8.18-8.14(m,2H),8.10-8.07(m,1H),7.82(s,1H),7.64-7.59(m,1H),7.47-7.44(m,3H),4.76-4.60(m ,1H),5.70-5.60(m,1H),3.87-3.52(m,6H),3.23-3.10(m,0.5H),2.7 8-2.67(m,0.5H),2.47(s,3H),2.28-2.20(m,1H),1.71-1.59(m,7H).

[0174] Example 3: (C89-P1, C89-P2, C89-P3)

[0175] 1) Synthesis of intermediate C89-2:

[0176] To a 1000 mL three-necked flask, C89-1 (9.0 g, 44.53 mmol) and anhydrous tetrahydrofuran (225 mL) were added sequentially. 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% w / w 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 C89-2 (3.5 g, 33%).

[0177] 2) Synthesis of intermediate C89-3:

[0178] To a solution of C89-2 (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 C89-3 (2.8 g, 71%) as a white solid.

[0179] 3) Synthesis of intermediate C89-4:

[0180] C89-3 (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 C89-4 (300 mg, mixture, yield: 98.8%) as a brown oil. LCMS (ESI) m / z = 208.1 [M+H] + .

[0181] 4) Synthesis of intermediate C89-5:

[0182] Compound C87-6 (240 mg, 0.77 mmol) was added to dichloromethane (10 mL), and CMPI (217.16 mg, 0.85 mmol), DIEA (299.61 mg, 2.32 mmol), and C89-4 (281.82 mg, 1.16 mmol) were added at 0°C. The reaction solution was heated to 50°C and allowed to react for 2 hours. LC-MS showed that the starting material was completely reacted. Aqueous solution was slowly added dropwise to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (DCM / MeOH = 100 / 1) to obtain compound C89-5 (180 mg, 47% yield) as a yellow solid. LCMS (ESI) m / z = 500.1 [M+H] + .

[0183] 5) Synthesis of intermediate C89-6:

[0184] C89-5 (180 mg, 0.36 mmol) was added to dichloromethane (15 mL), and NBS (128.16 mg, 0.72 mmol) was added at 0°C. The mixture was then heated to 50°C and stirred overnight. LC-MS showed that the reaction was complete. Aqueous solution was added dropwise to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (DCM / MeOH = 100 / 1) to afford C89-6 (160 mg, 77% yield) as a pale yellow solid. LCMS (ESI) m / z = 579.8 [M+H] + .

[0185] 6) Synthesis of intermediate C89-7:

[0186] Compound C89-6 (160 mg, 0.28 mmol), compound C88-1 (464.25 mg, 4.15 mmol), and silver tetrafluoroborate (109.2 mg, 0.56 mmol) were added to dimethyl sulfoxide (4 mL). The reaction solution was heated to 120° C. and reacted for 2 hours. LC-MS showed that the starting materials were completely reacted. Aqueous solution was added dropwise to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (DCM / MeOH / H 2 O.NH 3 = 10 / 1 / 0.01) to obtain compound C89-7 (100 mg, 59% yield) as an off-white solid. LCMS (ESI) m / z = 610.1 [M+H] + .

[0187] 7) Synthesis of C89-P1, C89-P2, and C89-P3:

[0188] Compound C87-11 (34.5 mg, 0.23 mmol) was added to DMF (5 mL), and HOBT (23.76 mg, 0.18 mmol) and EDCI (46.08 mg, 0.24 mmol) were added at room temperature. The mixture was stirred at room temperature for 2 hours under nitrogen protection. Then, compound C89-7 (100 mg, 0.16 mmol) and DIEA (61.92 mg, 0.48 mmol) were added. The nitrogen atmosphere was replaced three times and stirred at room temperature for 4 hours under nitrogen protection. LC-MS showed that the raw material reaction was complete. The reaction solution was concentrated under reduced pressure and the residue was purified by preparative high performance liquid chromatography (RP-PREP-3 SunFire C18 5um 19*150mm 18min-55-70B, A:H2O (0.2% FA), B: ACN, UV: 214 nm, flow rate 15 ml / min) to give white solids C89-P1 (10.5 mg, yield 8.6%, retention time 10.52 minutes), C89-P2 (11.2 mg, yield 9.2%, retention time 11.31 minutes), and C89-P3 (16.6 mg, yield 13.6%, retention time 12.56 minutes). C89-P1 and C89-P2 are both single-configuration compounds, while C89-P3 is a mixture of two diastereomers.

[0189] C89-P1 (the configuration was confirmed by crystal structure, C89-P1 is compound C2): LCMS (ESI) m / z = 744.1 [MH] - . 1 HNMR (400MHz, DMSO_d6): δ10.49(brs,1H),8.48(s,1H),8.10-8.08(m,2H),7.53-7.52(m,3H),7.43(d,J=8.8Hz,1H),7.35(d,J=8.8Hz,1H),5.5 9-5.58(m,1H),4.45-4.40(m,1H),3.71-3.69(m,2H),3.08-2.71(m,5H) ,2.41(s,3H),2.09-2.01(m,1H),1.68-1.63(m,1H),1.60-1.51(m,6H).

[0190] C89-P2: LCMS(ESI)m / z=744.1[MH] - . 1H NMR (400MHz, DMSO_d6): δ10.54(brs,1H),8.54(s,1H),8.11-8.09(m,2H),7.54-7.53(m,3H),7.44(d,J=8.8Hz,1H),7.35(d,J=8.8Hz,1H),5.4 8-5.46(m,1H),4.43-4.39(m,1H),3.72-3.71(m,2H),3.08-3.05(m,5H) ,2.43(s,3H),2.02-1.96(m,1H),1.68-1.61(m,1H),1.60-1.51(m,6H).

[0191] C89-P3: LCMS(ESI)m / z=744.1[MH] - . 1 H NMR (400MHz, DMSO_d6): δ10.53(brs,1H),8.51(s,1H),8.11-8.09(m,2H),7.54-7.53(m,3H),7.42(d,J=8.8Hz,1H),7.33(d,J=8.8Hz ,1H),5.56-5.53(m,1H),4.46-4.43(m,1H),3.81-3.69(m,2H),3.09-3.01(m,5H),2.42(s,3H),2.03-2.01(m,1H),1.64-1.53(m,6H).

[0192] Example 4: (C90-P1, C90-P2, C90-P3)

[0193] Referring to the synthesis method of compounds C87-P1 and C87-P2 in Example 1, C90 was synthesized by substituting C89-4 for C87-7. C90 was subjected to chiral resolution (IBN, MeOH:EtOH:FA = 50:50:0.3, 25 ml / min, 254 nm) to afford C90-P1 (2.0 mg, retention time 9.55 minutes), C90-P2 (2.3 mg, retention time 23.31 minutes), and C90-P3 (2.1 mg, retention time 32.67 minutes). C90-P1 was a mixture of two diastereomers, while C90-P2 and C90-P3 were both single-configuration compounds.

[0194] C90-P1: LCMS(ESI)m / z=720.1[M+H] + . 11H NMR (400 MHz, DMSO-d6): δ 10.48 (brs, 1H), 8.56 (s, 1H), 8.12 - 8.08 (m, 2H), 7.55 - 7.52 (m, 3H), 7.44 (d, J = 8.8 Hz, 1H), 7.36 (d, J = 8.8 Hz, 1H), 5.48 - 5.46 (m, 1H), 3.77 - 3.73 (m, 1H), 3.08 - 3.02 (m, 6H), 2.51 - 2.50 (m, 1H), 2.46 - 2.44 (m, 4H), 2.01 - 1.99 (m, 2H), 1.53 - 1.44 (m, 3H).

[0195] C90 - P2: LCMS (ESI) m / z = 720.1 [M + H] + 。 1 1H NMR (400 MHz, DMSO-d6): δ 10.50 (brs, 1H), 8.49 (s, 1H), 8.33 (brs, 1H), 8.11 - 8.09 (m, 2H), 7.53 - 7.52 (m, 3H), 7.43 (d, J = 8.8 Hz, 1H), 7.36 (d, J = 8.8 Hz, 1H), 5.56 - 5.33 (m, 1H), 3.78 - 3.72 (m, 1H), 3.15 - 2.94 (m, 6H), 2.75 - 2.66 (m, 2H), 2.47 (s, 3H), 2.03 - 1.96 (m, 2H), 1.52 - 1.46 (m, 3H).

[0196] C90 - P3: LCMS (ESI) m / z = 720.1 [M + H] + 。 1 1H NMR (400 MHz, DMSO-d6): δ 10.49 (brs, 1H), 8.47 (s, 1H), 8.35 (brs, 1H), 8.12 - 8.10 (m, 2H), 7.54 - 7.53 (m, 3H), 7.44 (d, J = 8.8 Hz, 1H), 7.34 (d, J = 8.8 Hz, 1H), 5.49 - 5.47 (m, 1H), 3.77 - 3.75 (m, 1H), 3.08 - 2.97 (m, 6H), 2.53 - 2.49 (m, 2H), 2.41 (s, 3H), 2.03 - 1.96 (m, 2H), 1.46 - 1.44 (m, 3H).

[0197] Example 5: (C93 - P1, C93 - P2, C93 - P3, C93 - P4)

[0198] Referring to the synthesis method of Example 4, C93-1 was used instead of C87-3, and C88-1 was used instead of piperazine. The final step was purified by preparative high performance liquid chromatography (Waters-PREP-8SunFire C18 5um 19*150mm 18min-45-55B, A:H2O (0.2% FA), B:ACN, UV:214nm, flow rate 15ml / min) to obtain C93-P1 (1.4mg, yield 2.9%, retention time 9.83min), C93-P2 (1.9mg, yield 3.9%, retention time 10.70min), C93-P3 (1.6mg, yield 3.3%, retention time 11.57min) and C93-P4 (1.2mg, yield 2.5%, retention time 12.03min).

[0199] C93-P1:LCMS(ESI)m / z=747.0[M+H] + . 1 H NMR (400MHz, CD3OD): δ8.69(d,J=4.4Hz,1H),8.53(s,1H),8.30(d,J=8.0Hz,1H),8.00-7.96(m,1H),7.55-7.48(m,2H),7.18(d,J=8.8H z,1H),5.64-5.61(m,1H),5.35-5.33(m,1H),3.35-3.33(m,7H),2.80-2.76(m,1H),2.53(s,3H),2.03-2.01(m,1H),1.79-1.65(m,6H).

[0200] C93-P2:LCMS(ESI)m / z=747.0[M+H] + . 1 H NMR (400MHz, CD3OD): δ8.69(d,J=3.6Hz,1H),8.48(s,1H),8.30(d,J=7.6Hz,1H),8.00-7.96(m,1H),7.55-7.52(m,1H),7.46(d,J=8.8Hz,1H),7.1 9(d,J=8.8Hz,1H),5.58-5.55(m,1H),5.35-5.33(m,1H),3.35-3.33(m,7 H),2.48(s,3H),2.37-2.36(m,1H),2.03-2.01(m,1H),1.52-1.50(m,6H).

[0201] C93-P3:LCMS(ESI)m / z=747.0[M+H] + .1 1H NMR (400 MHz, CD3OD): δ 8.69 (d, J = 4.4 Hz, 1H), 8.48 (s, 1H), 8.31 (d, J = 8.0 Hz, 1H), 8.00 - 7.97 (m, 1H), 7.55 - 7.52 (m, 1H), 7.47 (d, J = 8.8 Hz, 1H), 7.20 (d, J = 8.8 Hz, 1H), 5.61 - 5.58 (m, 1H), 5.35 - 5.33 (m, 1H), 3.32 - 3.31 (m, 7H), 2.49 (s, 3H), 2.34 - 2.31 (m, 1H), 2.03 - 1.97 (m, 1H), 1.58 - 1.56 (m, 6H).

[0202] C93 - P4: LCMS (ESI) m / z = 747.0 [M + H] + . 1 1H NMR (400 MHz, CD3OD): δ 8.60 (d, J = 3.2 Hz, 1H), 8.43 (s, 1H), 8.21 (d, J = 7.6 Hz, 1H), 7.91 - 7.87 (m, 1H), 7.45 - 7.42 (m, 1H), 7.38 (d, J = 8.8 Hz, 1H), 7.07 (d, J = 8.4 Hz, 1H), 5.55 - 5.52 (m, 1H), 5.25 - 5.23 (m, 1H), 3.32 - 3.31 (m, 7H), 2.66 - 2.69 (m, 1H), 2.41 (s, 3H), 1.94 - 1.92 (m, 1H), 1.57 - 1.49 (m, 6H).

[0203] Example 6: (C43 - P1, C43 - P2, C43 - P3)

[0204] Referring to the synthetic method of Example 3, C43-1 was used instead of C89-4, and the final step was purified by preparative high performance liquid chromatography (Waters-PREP-8SunFire C18 5um 19*150mm 18min-42-78B, A:H2O (0.2% FA), B:ACN, UV:214nm, flow rate 15ml / min) to give C43-P1 (11.1mg, yield 5.7%, retention time 11.82min), C43-P3 (14.1mg, yield 7.3%, retention time 13.25min) and C43-P2 crude product (30mg, retention time 12.82min). The crude C43-P2 was purified by chiral separation (IBN, Hex:EtOH:DEA=50:50:0.3, 25 ml / min, 230 nm) to give C43-P2 (12 mg, yield 6.2%, retention time 7.997 min).

[0205] C43-P1 (hydrochloride salt, single configuration compound): 1 H NMR (400MHz, CD3OD): δ8.80(s,1H),8.12-8.10(m,2H),7.68(s,1H),7.47- 7.41(m,3H),7.31-7.28(m,1H),7.18-7.16(m,1H),5.50-5.47(m,1H),4.62 -4.59(m,1H),4.00-3.44(m,6H),2.71-2.70(m,1H),2.64(s,3H),2.52-2.4 5(m,1H),1.83-1.62(m,6H),1.42-1.39(m,1H).LCMS(ESI)m / z=712.1[M+H] + .

[0206] C43-P2 (free base, single configuration compound): 1 H NMR (400MHz, DMSO-d6): δ10.86(s,1H),8.54(s,1H),8.05-8.03(m,2H),7.74(s,1 H),7.50-7.48(m,3H),7.41-7.39(m,1H),7.32-7.29(m,1H),5.36(d,J=9.2Hz,1H ),4.40-4.38(m,1H),3.71-3.70(m,2H),3.24-2.95(m,6H),2.43(s,3H),2.10(d, J=13.6Hz,1H),1.75-1.34(m,6H),1.23-1.17(m,1H).LCMS(ESI)m / z=712.1[M+H] + .

[0207] C43-P3 (hydrochloride salt, mixture of two diastereomers): 1 H NMR (400MHz, CD3OD): δ8.86(s,1H),8.13-8.10(m,2H),7.68-7.66(m,1H),7. 47-7.41(m,3H),7.30-7.26(m,1H),7.18-7.15(m,1H),5.49-5.42(m,1H),4. 71-4.68(m,1H),3.79-3.54(m,6H),3.13-3.03(m,1H),2.69(s,3H),2.50-2. 43(m,1H),2.01-1.55(m,6H),1.40-1.37(m,1H).LCMS(ESI)m / z=712.1[M+H] + .

[0208] Example 7: (C142-P1, C142-P2, C142-P3-A, C142-P3-B)

[0209] 1) Synthesis of intermediate C142-2:

[0210] Compound C142-1 (0.6 g, 2.38 mmol, prepared according to the synthesis method disclosed in patent application WO2008082484) and methylboronic acid (1.42 g, 23.8 mmol) were dissolved in a mixture 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) dichloride (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 C142-2 (0.4 g, yield: 89.8%) as a white solid. m / z [M+H] + =188.1.

[0211] 2) Synthesis of products:

[0212] The synthetic method of reference Example 3 was used, but C142-2 was substituted for C89-4. The final step was purified by preparative high-performance liquid chromatography (RP-PREP-3 SunFire C18 5 μm 19*150 mm 18 min-50-75 B, A: H2O (0.2% FA), B: ACN, UV: 214 nm, flow rate 15 ml / min) to obtain C142-P1 (33.6 mg, yield 12.4%, retention time 10.13 min), C142-P2 (25.2 mg, yield 9.3%, retention time 10.74 min), and C142-P3 (54.1 mg, yield 19.9%, retention time 11.53 min). C142-P1 and C142-P2 were both single-configuration compounds, while C142-P3 was a mixture of two diastereomers. C142-P3 was subjected to chiral separation (IBN, Hex:EtOH:TFA=50:50:0.2, 25 ml / min, 254 nm) to give C142-P3-A (10.7 mg, retention time 11.1 min) and C142-P3-B (14.1 mg, retention time 37.7 min).

[0213] C142-P1: LCMS(ESI)m / z=726.1[M+H] + . 1 H NMR (400MHz, DMSO_d6): δ10.24(s,1H),8.53(s,1H),8.10-8.02(m,2H),7.56-7.48(m,3H ),7.23(d,J=8.8Hz,1H),7.10(d,J=8.8Hz,1H),5.49-5.45(m,1H),4.40-4.38(m,1H),3. 76-3.73(m,2H),3.35-3.17(m,4H),2.67-2.52(m,1H),2.51-2.47(m,1H),2.45(s,3H),2 .22(s,3H),1.67-1.66(m,1H),1.65-1.57(m,3H),1.49-1.42(m,2H),1.23-1.22(m,2H).

[0214] C142-P2: LCMS(ESI)m / z=726.1[M+H] + . 1H NMR (400MHz, DMSO_d6): δ10.28(s,1H),8.50(s,1H),8.10-8.08(m,2H),7.54-7.52( m,3H),7.24(d,J=8.4Hz,1H),7.10(d,J=8.4Hz,1H),5.41-5.38(m,1H),4.40-4.38( m,1H),3.74-3.72(m,2H),3.31-3.23(m,4H),3.04-3.01(m,1H),2.42(s,3H),2.23( s,3H),2.41-2.11(m,1H),1.68-1.66(m,1H),1.51-1.40(m,5H),1.23-1.22(m,2H).

[0215] C142-P3-A: LCMS(ESI)m / z=726.1[M+H] + . 1 H NMR (400MHz, DMSO_d6): δ10.28(s,1H),8.56(s,1H),8.12-8.05(m,2H),7.58-7. 50(m,3H),7.25-7.21(m,1H),7.11(d,J=8.4Hz,1H),5.50-5.45(m,1H),4.45-4. 42(m,1H),3.76-3.71(m,1H),3.61-3.45(m,5H),2.63-2.58(m,1H),2.50-2.35( m,4H),2.21(s,3H),1.66-1.63(m,1H),1.49(d,J=6.8Hz,3H),1.44-1.16(m,4H).

[0216] C142-P3-B (the configuration was confirmed by crystal structure analysis; C142-P3-B is compound C100): LCMS (ESI) m / z = 726.1 [M+H] + . 1H NMR (400MHz, DMSO_d6): δ10.26(s,1H),8.56(s,1H),8.12-8.05(m,2H),7.58-7.50(m, 3H),7.24(d,J=8.4Hz,1H),7.08(d,J=8.4Hz,1H),5.41(d,J=8.4Hz,1H),4.47-4.42(m, 1H),3.80-3.73(m,2H),3.61-3.45(m,4H),3.10-3.02(m,1H),2.44(s,3H),2.22(s,3H ),2.10(d,J=13.6Hz,1H),1.68-1.63(m,1H),1.48(d,J=6.8Hz,3H),1.44-1.16(m,4H).

[0217] Example 8: (C143-P1, C143-P2, C143-P3, C143-P4)

[0218] Referring to the synthetic method of Example 5, C142-2 was used instead of C89-4, and the final step was purified by preparative high performance liquid chromatography (Waters-PREP-8SunFire C18 5um 19*150mm 18min-45-50B, A:H2O (0.2% FA), B:ACN, UV:214nm, flow rate 15ml / min) to obtain C143-P1 (2.4mg, yield 3%, retention time 9.88min), C143-P2 (2.1mg, yield 2.6%, retention time 10.25min), C143-P3 (3.6mg, yield 4.5%, retention time 12.07min) and C143-P4 (3.6mg, yield 4.5%, retention time 12.55min).

[0219] C143-P1: LCMS(ESI)m / z=727.1[M+H] + . 1H NMR(400MHz,DMSO_d6):δ10.28(s,1H),8.73(d,J=4.0Hz,1H),8.48(s,1H),8.14(d,J=8.0Hz,1H),8.01-7.97(m,1H),7.55-7.52(m,1H),7.23-7.20(m,1H),7.16-7.13(m,1H),5.51-5.47(m,1H),4.48-4.46(m,1H),3.76-3.64(m,2H),2.99-2.96(m,4H),2.67-2.58(m,1H),2.33(s,3H),2.21(s,3H),2.03-1.97(m,1H),1.55-1.35(m,7H).

[0220] C143-P2:LCMS(ESI)m / z=727.1[M+H] + 。 1 H NMR(400MHz,DMSO_d6):δ10.33(s,1H),8.72(d,J=4.0Hz,1H),8.47(s,1H),8.14(d,J=8.0Hz,1H),8.01-7.96(m,1H),7.54-7.51(m,1H),7.24-7.20(m,1H),7.16-7.12(m,1H),5.50-5.44(m,1H),4.41-4.40(m,1H),3.73-3.64(m,2H),3.17-3.05(m,4H),2.44(s,3H),2.40-2.39(m,1H),2.27(s,3H),2.03-1.89(m,1H),1.59-1.45(m,7H).

[0221] C143-P3:LCMS(ESI)m / z=727.1[M+H] + 。 1H NMR (400MHz, DMSO_d6): δ10.32(s,1H),8.73(d,J=4.4Hz,1H),8.43(s,1H),8.15(d, J=8.0Hz,1H),8.00-7.96(m,1H),7.55-7.52(m,1H),7.25-7.23(m,1H),7.15-7.11( m,1H),5.46-5.44(m,1H),4.45-4.40(m,1H),3.80-3.64(m,2H),3.17-3.01(m,4H), 2.46-2.44(m,1H),2.34(s,3H),2.21(s,3H),2.03-1.90(m,1H),1.53-1.40(m,7H).

[0222] C143-P4: LCMS(ESI)m / z=727.1[M+H] + . 1 H NMR (400MHz, DMSO_d6): δ10.28(s,1H),8.74(d,J=4.0Hz,1H),8.49(s,1H),8.14(d, J=8.0Hz,1H),8.01-7.97(m,1H),7.55-7.52(m,1H),7.23-7.20(m,1H),7.16-7.13( m,1H),5.51-5.47(m,1H),4.45-4.43(m,1H),3.76-3.67(m,2H),3.10-2.93(m,4H), 2.64-2.59(m,1H),2.41(s,3H),2.20(s,3H),2.03-1.97(m,1H),1.56-1.34(m,7H).

[0223] Example 9: (C144-P1, C144-P2, C144-P3-A, C144-P3-B)

[0224] 1) Synthesis of intermediate C144-2:

[0225] Compound C144-1 (50 g, 341.9 mmol) was added to acetonitrile (500 mL), followed by a 2 mol / L dimethylamine tetrahydrofuran solution (200 mL). The mixture was reacted at 50°C for 4 hours. Hydrazine hydrate (51.35 g, 820.6 mmol) was then added, and the mixture was refluxed overnight. LCMS indicated the reaction was complete. The reaction mixture was returned to room temperature and concentrated under reduced pressure to a residual volume of 230 mL. Solid precipitated, which was filtered to obtain a filter cake. The filter cake was dried to yield C144-2 (39 g, yield: 89.7%). m / z [M+H] +=128.0.

[0226] 2) Synthesis of intermediate C144-3:

[0227] Compound C144-2 (32 g, 251.7 mmol) was added to acetic acid (210 mL), followed by methyl propionylacetate (39.3 g, 302.0 mmol). The mixture was allowed to react at 90°C overnight. LCMS indicated the reaction was complete. The reaction mixture was returned to room temperature and concentrated under reduced pressure to remove the acetic acid. Ethyl acetate (180 mL) and petroleum ether (110 mL) were then added and triturated with stirring for 1 hour. The resulting filter cake was filtered and dried to afford C144-3 (50 g, yield: 95.9%). m / z [M+H] = 208.1.

[0228] 3) Synthesis of intermediate C144-4:

[0229] Compound C144-3 (4.5 g, 21.72 mmol) was added to tetrahydrofuran (110 mL) and 1,4-dioxane (110 mL). LDA (2 M, 14 mL) was added under nitrogen and allowed to react at room temperature for 30 minutes. LDA (2 M, 14 mL) was added again at 0°C and stirred at 0°C for 30 minutes. Stirring continued at room temperature for 30 minutes. S-epichlorohydrin (5.22 g, 56.47 mmol) was added at room temperature and stirred overnight at room temperature. LCMS indicated the reaction was complete. The reaction solution was added to aqueous ammonium chloride and separated to obtain an organic phase. The aqueous phase was extracted twice with dichloromethane. The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography to afford C144-4 (4 g, yield: 69.96%). m / z [M+H] + =264.2.

[0230] 4) Synthesis of intermediate C144-5:

[0231] Compound C144-4 (3.5 g, 13.29 mmol) was added to dichloromethane (40 mL), and NBS (2.60 g, 14.62 mmol) was added at 0°C. The mixture was allowed to react at room temperature for 6 hours. LCMS indicated the reaction was complete. The reaction solution was diluted with dichloromethane and 10% sodium sulfite solution was added. The mixture was separated and extracted twice with dichloromethane. The organic phases were combined and washed twice with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to afford C144-5 (4 g, yield: 87.94%). m / z [M+H]+ = 342.1.

[0232] 5) Synthesis of intermediate C144-6:

[0233] Compound C144-5 (4 g, 11.69 mmol) was added to dichloromethane (80 mL), and an aqueous solution (100 mL) of sodium bicarbonate (9.82 g, 116.90 mmol), potassium bromide (0.14 g, 1.17 mmol), trioctylmethylammonium chloride (0.24 g, 0.58 mmol), and TEMPO (0.091 g, 0.58 mmol) were added. A 10% mass fraction of sodium hypochlorite aqueous solution (43.51 g, 58.45 mmol) was added at 0°C, and the reaction was carried out at room temperature for 2.5 hours. LCMS showed the reaction was complete. The reaction solution was added with dichloromethane, and the liquid was separated after extraction to obtain an aqueous phase. The aqueous phase was further extracted with dichloromethane. The organic phases were combined, and 10% aqueous sodium bicarbonate solution was added to the combined organic phases. The aqueous phase was extracted to obtain an aqueous phase. The two aqueous phases were combined, and dichloromethane was added to the aqueous phase. The pH was adjusted to 3-4 with dilute hydrochloric acid. The dichloromethane phases were extracted three times. The dichloromethane phases were combined, washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure to obtain C144-6 (2.5 g, yield: 60.05%). m / z [M+H] + = 356.1.

[0234] 6) Synthesis of intermediate C144-7:

[0235] Compound C144-6 (2.15 g, 6.04 mmol) was added to dichloromethane (40 mL), followed by 2-chloro-1-methylpyridinium iodide (1.70 g, 6.64 mmol), DIEA (3.51 g, 27.18 mmol), and C142-2 (1.76 g, 7.85 mmol). The mixture was stirred at room temperature for 4 hours. LCMS indicated the reaction was complete. Dichloromethane and water were added to the reaction mixture, and the mixture was separated and extracted twice with dichloromethane. The organic phases were combined, washed twice with dilute hydrochloric acid, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. Column chromatography afforded C144-7 (2.6 g, yield: 82.0%). m / z [M+H]+ = 525.1.

[0236] 7) Synthesis of intermediate C144-8:

[0237] Compound C144-7 (700 mg, 1.33 mmol) was added to DMSO (3 mL), followed by C88-1 (0.75 g, 6.65 mmol) and DIPA (0.67 g, 6.65 mmol). Silver tetrafluoroborate (0.39 g, 2.00 mmol) was added under nitrogen protection, and the mixture was microwaved at 105°C under nitrogen protection for 2 hours. LCMS indicated the reaction was complete. The reaction solution was returned to room temperature, diluted with dichloromethane, filtered, and the filtrate was concentrated under reduced pressure and purified by reverse phase column chromatography to afford C144-8 (270 mg, yield: 36.41%). m / z [M+H]+ =557.2

[0238] 8) Synthesis of product C144:

[0239] Compound C144-8 (0.10 g, 0.66 mmol) and HOAT (0.10 g, 0.76 mmol) were added to acetonitrile (3 mL), followed by EDCI (0.19 g, 0.98 mmol), and stirred at room temperature for 1 hour. Compound C87-11 (270 mg, 0.49 mmol) and DIEA (0.22 g, 1.71 mmol) were then added, and the mixture was stirred at room temperature for 30 minutes. LCMS showed the reaction was complete. The reaction solution was filtered and the filtrate was purified by reverse phase column to give C144 (110 mg, yield: 32.74%). Preparative HPLC purification (SHIMADZU LC 20, Agilent 10Prep-C18, 250*21.2 mm, 10 μm, A: HO (0.1% FA), B: ACN, UV: 214 nm, flow rate 20 ml / min) gave C144-P1 (19.8 mg, yield 4.3%, retention time 13.1 min), C144-P2 (7.3 mg, yield 1.6%, retention time 14.0 min), and C144-P3 (retention time 16.2 min). C144-P1 and C144-P2 were both single-configuration compounds, while C144-P3 was a mixture of two diastereomers. C144-P3 was subjected to chiral separation (Waters SFC-150mgm, Daicel OJ, 30*250mm, 10um, CO2 / MEOH=75 / 25, 50ml / min, 254nm) to obtain C144-P3-A (8mg, retention time 12.01min) and C144-P3-B (19.7mg, retention time 16.19min).

[0240] C144-P1:LCMS m / z[M+H] + =693.3. 11H NMR (400 MHz, chloroform-d) δ 11.46 (s, 1H), 8.69 (s, 1H), 8.59 (s, 1H), 7.24 (s, 1H), 6.87 (d, J = 8.5 Hz, 1H), 5.20 (d, J = 8.2 Hz, 1H), 4.66 (s, 1H), 4.12 (d, J = 66.5 Hz, 2H), 3.70 (dd, J = 15.7, 6.7 Hz, 2H), 3.08 (s, 6H), 3.06–2.98 (m, 1H), 2.54 (s, 3H), 2.43–2.27 (m, 1H), 2.24–2.16 (m, 1H), 2.14 (s, 3H), 2.07–1.95 (m, 1H), 1.68–1.57 (m, 1H), 1.55–1.38 (m, 4H), 1.37–1.28 (m, 1H), 1.20–1.09 (m, J = 9.7 Hz, 1H).

[0241] C144-P2: LCMS m / z [M+H] + = 693.3。 1 1H NMR (400 MHz, chloroform-d) δ 11.46 (s, 1H), 8.69 (s, 1H), 8.59 (s, 1H), 7.24 (s, 1H), 6.87 (d, J = 8.5 Hz, 1H), 5.20 (d, J = 8.2 Hz, 1H), 4.66 (s, 1H), 4.12 (d, J = 66.5 Hz, 2H), 3.70 (dd, J = 15.7, 6.7 Hz, 2H), 3.08 (s, 6H), 3.​​​​​​H (400 MHz, chloroform-d) δ 11.49 (s, 1H), 9.20 (s, 1H), 8.59 (s, 1H), 7.22 (d, J = 8.6 Hz, 1H), 6.88 (d, J = 8.5 Hz, 1H), 5.23 (d, J = 9.1 Hz, 1H), 4.71 (s, 1H), 4.09 (s, 2H), 3.74–3.50 (m, 2H), 3.10 (s, 6H), 2.81 (d, J = 13.1 Hz, 1H), 2.68 (dd, J = 20.1, 8.8 Hz, 1H), 2.54 (s, 3H), 2.35 (q, J = 9.6 Hz, 1H), 2.16 (s, 3H), 2.06–1.97 (m, 1H), 1.68–1.57 (m, 1H), 1.55–1.38 (m, 4H), 1.37–1.28 (m, 1H), 1.20–1.09 (m, J = 9.7 Hz, 1H).

[0243] C144 - P3 - B: LCMS m / z [M + H] + = 693.3。 1 H (400 MHz, chloroform-d) δ 11.49 (s, 1H), 9.20 (s, 1H), 8.59 (s, 1H), 7.22 (d, J = 8.6 Hz, 1H), 6.88 (d, J = 8.5 Hz, 1H), 5.23 (d, J = 9.1 Hz, 1H), 4.71 (s, 1H), 4.09 (s, 2H), 3.74–3.50 (m, 2H), 3.10 (s, 6H), 2.81 (d, J = 13.1 Hz, 1H), 2.68 (dd, J = 20.1, 8.8 Hz, 1H), 2.54 (s, 3H), 2.35 (q, J = 9.6 Hz, 1H), 2.16 (s, 3H), 2.06–1.97 (m, 1H), 1.68–1.57 (m, 1H), 1.55–1.38 (m, 4H), 1.37–1.28 (m, 1H), 1.20–1.09 (m, J = 9.7 Hz, 1H).

[0244] Example 10: (C146 - P1, C146 - P2, C146 - P3 - A, C146 - P3 - B)

[0245] Referring to the synthesis method of Example 9, tetrahydropyrrole was used instead of dimethylamine hydrochloride in the first step. In the final step, the product was purified by preparative high-performance liquid chromatography (SHIMADZU LC 20, Agilent 10Prep-C18, 250*21.2 mm, 10 μm, A: H2O (0.1% FA), B: ACN, UV: 214 nm, flow rate 20 ml / min) to obtain C146-P1 (retention time 10.5 min), C146-P2 (retention time 11.3 min), and C146-P3 (retention time 12.6 min). C146-P1 and C146-P2 were both single-configuration compounds, while C146-P3 was a mixture of two diastereomers. C146-P3 was subjected to chiral separation (Waters SFC-150 mgm, Daicel OD, 30*250 mm, 10 um, CO2 / MEOH=65 / 35, 50 ml / min, 214 nm) to give C146-P3-A (5.81 min) and C146-P3-B (11.14 min).

[0246] C146-P1:LCMS m / z[M+H] + =719.3, 1 H NMR (400MHz, DMSO-d6) δ10.17(s,1H),8.54(s,1H),7.23(d,J=8.5Hz,1H),7.07(d,J=8.6Hz,1H),5.30(dq,J=8.3,5 .0Hz,1H),4.36(d,J=13.0Hz,1H),3.65(dq,J=22.0,7.2Hz,3H),3.54–3.43(m,4H),3.25–3.10(m,3H),2.96(dt,J=1 3.5,9.9Hz,1H),2.42(s,3H),2.35(ddd,J=12.6,8.1,5.9Hz,1H),2.19(s,3H),1.99(dt,J=13.0,7.0Hz,1H),1.90( t,J=5.9Hz,4H),1.61(d,J=8.5Hz,1H),1.51(d,J=7.1Hz,1H),1.39(dd,J=20.7,10.2Hz,3H),1.15(t,J=8.9Hz,1H).

[0247] C146-P2:LCMS m / z[M+H] + =719.3, 1H NMR(400MHz,DMSO-d6)δ10.17(s,1H),8.54(s,1H),7.23(dd,J=8.6,1.9Hz,1H),7.06(dd,J=11.0,8.6Hz,1H),5.31–5.22(m,1H),4.38(t,J=16.1Hz,1H),3.65(dq,J=22.0,7.2Hz,3H),3.54–3.43(m,4H),3.25–3.10(m,3H),2.96(dt,J=13.5,9.9Hz,1H),2.42(s,3H),2.35(ddd,J=13.2,8.8,6.2Hz,1H),2.19(s,3H),2.01(d,J=13.7Hz,1H),1.93–1.86(m,4H),1.61(d,J=8.5Hz,1H),1.51(d,J=7.1Hz,1H),1.39(dd,J=20.7,10.2Hz,3H),1.15(t,J=8.9Hz,1H)。

[0248] C146-P3-A:LCMS m / z[M+H] + =719.3, 1 H NMR(400MHz,DMSO-d6)δ10.19(s,1H),8.55(s,1H),7.22(d,J=8.5Hz,1H),7.07(d,J=8.6Hz,1H),5.31(dt,J=9.0,5.1Hz,1H),4.47–4.29(m,1H),3.66–3.57(m,3H),3.54–3.43(m,4H),3.25–3.10(m,3H),2.96(dt,J=13.5,9.9Hz,1H),2.42(s,3H),2.35(ddd,J=13.2,8.8,6.2Hz,1H),2.19(s,3H),1.99(dt,J=13.0,6.8Hz,1H),1.93–1.86(m,4H),1.61(d,J=8.5Hz,1H),1.51(d,J=7.1Hz,1H),1.39(dd,J=20.7,10.2Hz,3H),1.15(t,J=8.9Hz,1H)。

[0249] C146-P3-B:LCMS m / z[M+H] + =719.3, 1H NMR (400MHz, DMSO-d6) δ10.18(s,1H),8.55(s,1H),7.24(d,J=8.6Hz,1H),7.05(d,J=8.6Hz,1H),5.2 4(d,J=9.8Hz,1H),4.42(d,J=9.1Hz,1H),3.75–3.48(m,3H),3.54–3.43(m,4H),3.27–3.12(m,3H),3. 09–2.90(m,1H),2.43(s,3H),2.19(s,3H),2.04–1.98(m,1H),1.90(t,J=6.5Hz,4H),1.62(dd,J=15. 1,7.2Hz,1H),1.52–1.47(m,1H),1.42(d,J=7.3Hz,3H),1.34(d,J=8.4Hz,1H),1.16(t,J=8.6Hz,1H).

[0250] Example 11: (C145, C145-P1, C145-P2)

[0251] 1) Synthesis of intermediate C145-2:

[0252] Compound C145-1 (0.1 g, 0.19 mmol, prepared according to the synthesis method disclosed in patent application WO2024079623) was added to dichloromethane (2 mL), followed by 2-chloro-1-methylpyridinium iodide (0.053 g, 0.21 mmol), DIEA (0.11 g, 0.85 mmol), and C142-2 (0.55 g, 0.25 mmol), and stirred at room temperature for 4 hours. LCMS showed that the reaction was complete. Dichloromethane and water were added to the reaction solution, and the mixture was separated and extracted twice with dichloromethane. The organic phases were combined, washed twice with dilute hydrochloric acid, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. Column chromatography gave C145-2 (70 mg, yield: 54.0%). m / z [M+H] + =696.1.

[0253] 2) Synthesis of intermediate C145-3:

[0254] Compound C145-2 (0.7 g, 0.1 mmol) was dissolved in dichloromethane (1 mL) and trifluoroacetic acid (1 mL) was added dropwise. The reaction mixture was stirred at 25°C for 1 hour. LCMS showed the reaction was complete. The reaction mixture was filtered and concentrated to give crude yellow oil C145-3 (100 mg, trifluoroacetate salt, yield: 100%). m / z [M+H] + =596.3.

[0255] 3) Synthesis of product C145:

[0256] Compound C87-11 (23 mg, 0.15 mmol) and HOAT (22 mg, 0.16 mmol) were added to DMF (1 mL), followed by EDCI (35 mg, 0.18 mmol) and stirring at room temperature for 1 hour. C145-3 (0.10 g crude product, 0.1 mmol) and DIEA (39 mg, 0.3 mmol) were then added and stirred at room temperature for 30 minutes. LCMS indicated the reaction was complete. The reaction solution was filtered, and the filtrate was purified by reverse phase column chromatography to afford C145 (40 mg, yield: 54.5%). LCMS m / z [M+H] + =732.4. 1 H NMR (400MHz, DMSO-d6) δ10.18(d,J=8.7Hz,1H),8.55(s,1H),7.27–7.19(m,1H),7.06(d,J=8.3Hz,1H),6.80(dd,J=4.6,2 .6Hz,1H),5.35(ddd,J=21.9,9.5,4.3Hz,1H),4.40(t,J=15.3Hz,1H),4.26(q,J=2.9Hz,2H),3.86–3.65(m,4H),3.55–3.4 3(m,2H),3.17(s,2H),3.05–2.94(m,1H),2.54–2.50(m,1H),2.43(s,3H),2.42–2.38(m,1H),2.18(s,3H),2.06(dt,J=12 .9,2.6Hz,1H),1.62(d,J=9.9Hz,1H),1.53(d,J=6.7Hz,1H),1.45(d,J=7.0Hz,3H),1.37–1.30(m,1H),1.21–1.11(m,1H).

[0257] 4) Synthesis of products C145-P1 and C145-P2:

[0258] C145 was purified by preparative high-performance liquid chromatography (SHIMADZU LC 20, Agilent 10Prep-C18, 250*21.2 mm, 10 μm, A: H2O (0.1% FA), B: ACN, UV: 214 nm, flow rate 20 ml / min) to give C145-P1 (8 mg, retention time 10.675 min) and C145-P2 (18 mg, retention time 11.147 min).

[0259] C145-P1:LCMS m / z[M+H] += 732.4. 1 1H NMR (400 MHz, DMSO-d6) δ 10.16 (s, 1H), 8.55 (s, 1H), 7.23 (d, J = 8.6 Hz, 1H), 7.08 (d, J = 8.6 Hz, 1H), 6.85–6.72 (m, 1H), 5.38 (dd, J = 8.5, 5.7 Hz, 1H), 4.37 (d, J = 11.3 Hz, 1H), 4.31–4.22 (m, 2H), 3.81 (t, J = 5.7 Hz, 2H), 3.74–3.65 (m, 2H), 3.56–3.45 (m, 2H), 3.35–3.15 (m, 4H), 2.58 (q, J = 7.2 Hz, 1H), 2.43 (s, 3H), 2.38 (dd, J = 8.0, 5.3 Hz, 1H), 2.18 (s, 3H), 2.00 (dq, J = 13.0, 6.1 Hz, 1H), 1.71–1.60 (m, 1H), 1.54 (d, J = 7.0 Hz, 3H), 1.48–1.40 (m, 1H), 1.21–1.12 (m, 1H).

[0260] C145 - P2: LCMS m / z [M + H] + = 732.4. 1 1H NMR (400 MHz, DMSO-d6) δ 10.22 (s, 1H), 8.48 (s, 1H), 7.24 (d, J = 8.5 Hz, 1H), 7.06 (d, J = 8.6 Hz, 1H), 6.80 (s, 1H), 5.33 (d, J = 9.8 Hz, 1H), 4.40 (d, J = 13.1 Hz, 1H), 4.26 (d, J = 3.1 Hz, 2H), 3.85–3.65 (m, 4H), 3.51 (dd, J = 16.8, 7.4 Hz, 2H), 3.20–3.10 (m, 4H), 3.07–2.93 (m, 1H), 2.54–2.50 (m, 1H), 2.41 (s, 3H), 2.18 (s, 3H), 2.06 (d, J = 13.5 Hz, 1H), 1.63 (q, J = 6.6, 5.4 Hz, 1H), 1.45 (d, J = 7.2 Hz, 3H), 1.40–1.27 (m, 1H), 1.19–1.10 (m, 1H).

[0261] Example 12: (C147)

[0262] The compound (100 mg, 0.88 mmol) and HOAT (100 mg, 0.73 mmol) were added to DMF (1 mL), followed by EDCI (150 mg, 0.78 mmol) and stirring at room temperature for 1 hour. C142-5 (0.08 g, 0.14 mmol) and DIEA (78 mg, 0.6 mmol) were then added and stirred at room temperature for 30 minutes. LCMS indicated the reaction was complete. The reaction solution was filtered, and the filtrate was purified on a reverse phase column to afford C147 (25 mg, yield: 26.5%). LCMS m / z [M+H]+ = 698.4. 1 H NMR (400MHz, DMSO-d6) δ10.30(d,J=9.2Hz,1H),8.21–8.03(m,3H),7.71(d,J=5.6Hz,1H),7.57(dd,J=5.2,2.0Hz, 3H),7.30(dd,J=23.9,8.6Hz,1H),7.14(dd,J=23.1,8.6Hz,1H),5.56–5.33(m,1H),4.47(t,J=7.4Hz,1H),3.89(s, 3H),3.80–3.69(m,1H),3.65–3.49(m,2H),3.43(d,J=5.3Hz,1H),3.08(dt,J=13.3,9.8Hz,1H),2.25(s,3H),2.15 (s,3H),2.11(t,J=2.6Hz,1H),1.60(d,J=7.0Hz,1H),1.50(d,J=7.2Hz,3H),1.46–1.36(m,1H),1.21–1.11(m,1H).

[0263] Biological testing section

[0264] Experimental Example 1 Biological Activity Test

[0265] ATPase activity assay of WRN helicase

[0266] 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.

[0267] 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.

[0268] 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.

[0269] Table 1 WRN ATPase activity data of the compounds in the examples of this application

[0270] 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.

[0271] Tumor cell proliferation inhibitory activity assay

[0272] 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.

[0273] Human colon adenocarcinoma SW48 cells were cultured in 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 in 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 in 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 by trypsinization.

[0274] After the test compounds were treated with the microsatellite unstable SW48 cell line and HCT116 cell line as well as the control cell DLD1-WRN-KO cell line for 4 days, the ATP level was detected using the CellCounting-Lite kit from Novozymes to evaluate the inhibitory effect of the test compounds on the growth of tumor cell lines.

[0275] 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.

[0276] Table 2 Tumor cell proliferation inhibitory activity data of the compounds in the examples of this application

[0277] 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.

[0278] Experimental Example 2 Liver microsome stability test

[0279] 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.

[0280] Experimental system:

[0281] 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.

[0282] Experiment Introduction:

[0283] 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).

[0284] Experimental methods:

[0285] 1. Preparation of Buffer

[0286] 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.

[0287] 2. Preparation of working solution

[0288] 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.

[0289] The control compounds testosterone, diclofenac, and propafenone were prepared as 10 mM stock solutions in DMSO and then further diluted in appropriate organic solvents.

[0290] 3. Preparation of Liver Microsome Solution

[0291] 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.

[0292] 4. Preparation of reduced nicotinamide adenine dinucleotide phosphate (NADPH) regeneration system

[0293] 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.

[0294] 5. Preparation of Stop Solution

[0295] The stop solution is prepared with acetonitrile or other organic solvent containing an internal standard (tolbutamide or other suitable compound) and stored in a refrigerator at 2-8°C.

[0296] 6. Incubation Process

[0297] 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.

[0298] 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.

[0299] 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.

[0300] 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.

[0301] Sample analysis

[0302] 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.

[0303] The CV of the internal standard peak area in each matrix should be within 20% for each analytical run.

[0304] Data Analysis

[0305] 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:

[0306] CL int(mic) =0.693 / T 1 / 2 / Microsomal protein content (microsomal concentration during incubation mg / mL)

[0307] CL int(liver) =CL int(mic) × amount of microsomal protein in the liver (mg / g) × liver weight to body weight ratio

[0308] According to the well stir model, the hepatic intrinsic clearance and hepatic clearance can be converted using the following formula.

[0309] CL (liver) =(CL int(liver) *Q h ) / (CL int(liver) +Q h )

[0310] The parameters in the formula are shown in the table below.

[0311] Parameters in data analysis formulas

[0312] Experimental Example 3 Hepatocyte Metabolic Stability Test

[0313] This experiment was used to test the metabolic stability of compounds in hepatocytes.

[0314] 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.

[0315] Experimental Example 4 Pharmacokinetic Test in Rats

[0316] In this experiment, the pharmacokinetic behavior of the test compound was investigated in SD rats after intravenous (IV) and oral (PO) administration.

[0317] 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.

[0318] 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.

[0319] Experimental Example 5 Pharmacokinetic Test in Mice

[0320] In this experiment, the pharmacokinetic behavior of the test compound was investigated in BALB / c mice after intravenous (IV) and oral (PO) administration.

[0321] 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 was administered with a single oral gavage. Whole blood samples were collected at the specified time (0.083, 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.

[0322] 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.

[0323] Experimental Example 6 hERG inhibition test

[0324] 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.

[0325] 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 Calculate the mean and standard error for each concentration 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.

[0326] Experimental Example 7 Cytochrome oxidase P450 inhibition test

[0327] 1) Preparation of buffer solution:

[0328] 100mM K-Buffer: Mix 9.5mL of stock solution A with 40.5mL of stock solution B, adjust the total volume to 500mL with ultrapure water, and titrate the buffer to pH 7.4 with KOH or H3PO4.

[0329] Raw material A (1M potassium dihydrogen phosphate): 136.5 g potassium dihydrogen phosphate in 1 L water;

[0330] Stock B (1M KH2PO4): 174.2 g KH2PO4 in 1 L water.

[0331] 2) Preparation of test substance

[0332] 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.

[0333] 3) In vitro incubation

[0334] 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.

[0335] 4) Detection of parent drug or metabolites

[0336] The concentration of parent drug or its metabolites in the incubation solution was determined by LC-MS / MS.

[0337] Experimental Example 8 Mouse Tumor Pharmacodynamic Model

[0338] In this experimental example, the in vivo efficacy of the test compound was evaluated in a mouse xenograft tumor model after oral administration (PO).

[0339] Human colon adenocarcinoma cell line SW48 was cultured in a monolayer in vitro using DMEM medium containing 10% fetal bovine serum, 1% penicillin, and streptomycin at 37°C and 5% CO2. The cells were 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 were subcutaneously inoculated on the right back of each mouse. On the 14th day after cell inoculation, the average tumor volume reached 200 mm 3 The mice were randomly divided into groups and given drugs by gavage once daily. Changes in body weight and tumor volume were recorded. After a certain number of days of dosing, the experiment was terminated. Changes in tumor volume and mouse body weight were statistically analyzed. The results showed that the compound described in this application has excellent tumor inhibitory effects.

[0340] Experimental Example 9 PXR activation test

[0341] In this experimental example, the compounds were tested for their activation effect on PXR.

[0342] 100 μL of DPX2 stably transfected cells (4.5×10 5 cells / mL, prepared with Puacyp medium), and then place the 96-well plate in a 37°C incubator. After 24 hours, remove the 96-well plate from the incubator, replace the medium with 100 μL of the test compound (final concentration of 10 μM and 30 μM), the positive control compound rifampicin (final concentration of 10 μM) and the DMSO treatment group (final concentration of 0.1%), and set up 3 replicates. Put the test plate back into the incubator. After 24 hours, replace the medium with freshly prepared test compound and rifampicin solution, and put the test plate back into the incubator again. After 48 hours of compound treatment, remove the test plate from the incubator, discard the medium in the wells, wash twice with PBS, and add 50 μL of 1x CellTiter-Fluor TM Incubate the plate with the cell viability assay reagent in a culture medium at 37°C for 30 minutes. Remove the test plate and equilibrate to room temperature. Measure the fluorescence signal using a microplate reader under the conditions of 400 nm excitation and 505 nm emission. Subsequently, add 50 μL of culture medium containing the ONE-Glo assay reagent to each well, vortex to mix, incubate at room temperature for 5 minutes, and measure chemiluminescence using a microplate reader. Analyze the data to evaluate the PXR activation activity of the test compound. The results show that the compound described in this application has no significant PXR activation activity.

[0343] Experimental Example 10: Test of saturated solubility of samples in FaSSIF solution

[0344] 1. Experimental Procedure

[0345] 1.1 Preparation of FaSSIF solution

[0346] Buffer solution (pH 6.5) preparation:

[0347] Weigh about 0.21 g of sodium hydroxide, about 2.24 g of sodium dihydrogen phosphate dihydrate, and 3.09 g of sodium chloride, add 500 ml of water, dissolve, and then adjust the pH to 6.5 with 1N sodium hydroxide or 1N hydrochloric acid.

[0348] Weigh 112 mg of FaSSIF solid into a 50 mL volumetric flask, add the above buffer to dissolve, dilute to the scale, shake well, and let stand at room temperature for more than 2 h.

[0349] 1.2 Sample preparation

[0350] Test sample: Take about 1 mg of the test sample, add 1 ml of FaSSIF solution, stir at room temperature overnight, centrifuge, and take the supernatant for analysis.

[0351] Reference solution: Accurately weigh about 1.5 mg of the test sample into a 50 mL volumetric flask, add DMF to dissolve and dilute to the scale, mix well to obtain the reference solution.

[0352] 1.3 Mobile phase preparation

[0353] Mobile phase A: Accurately measure 1000 mL of purified water, add 1 mL of formic acid, mix well, and degas by ultrasonication to obtain mobile phase A.

[0354] Mobile phase B: acetonitrile.

[0355] 1.4 Chromatographic conditions

[0356] Experimental Example 11: Co-crystal of small molecule and WRN protein

[0357] Purified recombinant WRN (500-945) protein (storage buffer: 50 mM HEPES, 500 mM NaCl, 0.5 mM TCEP, pH 7.5.5% glycerol) was concentrated to 7 mg / ml and mixed with the corresponding compound dissolved in 100% DMSO to a final concentration of 2 mM. After mixing, the mixture was incubated at 4°C for 2 hours and centrifuged at 12,000 rpm at 4°C for 10 minutes in a tabletop refrigerated centrifuge (Eppendorf 5418R). Crystallization was then performed on a Mosquito Xtal3 or Formulatrix NT8 pipetting robot. The crystallization kits used were commercial kits such as Hampton Research's Crystal Screen / Crystal Screen 2, Index, PEG / Ion, and PEGRx HT. The initial crystals obtained were optimized around the corresponding conditions, and finally diffraction-capable crystals were obtained under the conditions of 0.1M sodium citrate pH 5.0; 8-12% w / v PEG 6,000, or 0.2M potassium sodium tartrate; 0.1M BIS-TRIS pH 6.5; 8-12% w / v PEG 10,000, or 0.1M sodium citrate pH 5.0; 16-20% w / v PEG 20,000. The crystals were cryoprotected under crystallization conditions with 20%-25% glycerol, then quickly frozen in liquid nitrogen and sent to a synchrotron radiation source for data collection.

[0358] Experimental Example 12: CYP induction test

[0359] 1) Preparation and plating of human hepatocytes

[0360] Culture medium preparation

[0361] Prepare the following culture media in a biosafety cabinet and store at 4 °C until use:

[0362] Hepatocyte thawing medium (prepared by mixing the following components: Williams E medium, isotonic Percoll, DPBS, glutamine, HEPES, fetal bovine serum, human recombinant insulin, and dexamethasone).

[0363] Plating medium (prepared by mixing the following components: Williams E medium, fetal bovine serum, dexamethasone, penicillin / streptomycin, human recombinant insulin, glutamine, and HEPES).

[0364] Incubation medium (prepared by mixing the following components: Williams E medium, dexamethasone, ITS, penicillin / streptomycin, glutamine and HEPES, serum-free).

[0365] Hepatocyte thawing and processing

[0366] Thaw a vial of cryopreserved human hepatocytes in a 37°C water bath for 2 minutes. Wipe the vial with 70% alcohol in a biosafety cabinet. Use a wide-bore pipette tip to transfer the hepatocytes to 50 mL of prewarmed Hepatocyte Thawing Medium. Rinse the vial thoroughly by adding approximately 500 μL of Hepatocyte Thawing Medium, cap it, and invert it several times.

[0367] Centrifuge at 150g for 10 minutes at room temperature. Carefully aspirate the supernatant and dilute with plating medium to a seeding density of 0.55 × 10 6 cells / mL. Transfer 100 μL to each well of a 96-well plate coated with collagen I. Place the plate in an incubator at 37°C, 5% CO2 / 95% air, and 95% relative humidity for 4-6 hours.

[0368] Subsequent processing

[0369] After incubation, observe cell morphology under a microscope. Gently shake the plate to loosen debris and replace 125 μL of medium with 0.25 mg / mL Matrigel diluted in incubation medium. Return the plate to the incubator and incubate for an additional 18 hours. The culture is now ready for induction studies.

[0370] 2) Incubation with test compounds

[0371] Compound formulation

[0372] Test compounds were prepared at 1000× the maximum working concentration in DMSO and at the maximum working concentration in incubation medium. The solubility of the compound in both solutions was determined by visual inspection. Stock solutions of the test compound, negative control, and positive control inducer were then prepared at 1000× the final concentration in DMSO and diluted to their respective working concentrations using prewarmed incubation medium at 37°C. A negative control was prepared by adding 5 μL of DMSO to 5 mL of warmed incubation medium. Test compounds may sometimes be prepared at higher DMSO concentrations or directly in medium containing 0.1% DMSO to the final concentration.

[0373] Concentration of test compound and positive control inducer (rifampicin): 10 μM.

[0374] Processing Operations

[0375] Remove the hepatocyte plate from the incubator. Observe cell morphology under a microscope. Replace the culture medium in the corresponding wells with DMSO control, inducer, or test compound solution. Set up three replicates for each treatment.

[0376] Incubation and medium change

[0377] After 24 and 48 hours, the hepatocyte plates were removed from the incubator and the cell morphology was observed under a microscope. The medium was replaced with the test compound freshly diluted from the DMSO stock solution. The plates were returned to the incubator. The total incubation time was 72 hours.

[0378] 3) mRNA preparation and RT-PCR

[0379] mRNA preparation

[0380] mRNA was prepared and measured using the Cells-to-Ct kit purchased from Life Technologies. Remove any remaining CellTiter Cell Viability Assay Reagent, wash the cell monolayer twice with 125 μL of phosphate-buffered saline, and place the plate on ice. Add DNase to the lysis buffer as directed. Add 50 μL of lysis buffer to each well of the hepatocyte plate and mix the lysis reaction by pipetting up and down five times. Incubate the lysis reactions at room temperature for 8 minutes, then add 5 μL of stop solution to each lysis reaction and mix by pipetting up and down five times. Incubate at room temperature for an additional 2 minutes. Lysates can be stored at -20°C or -80°C for up to 5 months prior to RT.

[0381] Reverse transcription reaction

[0382] Program the QPCR system for reverse transcription: reverse transcription (hold), 1 cycle, 37°C, 60 min; RT inactivation (hold), 1 cycle, 95°C, 5 min; hold, 1 cycle, 4°C, indefinite.

[0383] Prepare a mixture for 106 reactions in a 15 mL tube and dispense into the corresponding wells of a 96-well PCR plate, preparing one tube of mixture per plate.

[0384] Reverse transcriptase master mix: 2× RT buffer, 25 μL per reaction; 20× RT enzyme mix, 2.5 μL per reaction; nuclease-free water, 7.5 μL per reaction; final volume of reverse transcription master mix: 35 μL.

[0385] Add 15 μL of sample lysate to each reverse transcription master mix aliquot, bringing the final reaction volume to 50 μL. A negative control (NC) was prepared by adding 15 μL of the previous mixture (not incubated with cells). After assembly, gently mix the reaction and briefly centrifuge to collect the contents at the bottom of the reaction vessel.

[0386] Incubate the samples at 37°C for 60 minutes using a QPCR system, followed by incubation at 95°C for 5 minutes to inactivate the RT enzyme. Store the prepared RT samples at -20°C until the QPCR reaction is performed.

[0387] Real-time PCR cycling

[0388] The QPCR system was programmed for real-time PCR cycles: enzyme activation (hold), 1 cycle at 95°C for 5 minutes, PCR (cycling), 45 cycles at 95°C for 15 seconds and 60°C for 1 minute.

[0389] Separate PCR cocktails were prepared for CYP 3A4; each cocktail contained a CYP-specific probe set and an ACTB probe set as an endogenous control gene.

[0390] PCR cocktail: Taqman gene expression master mix (2×), 10 μL per reaction; Taqman gene expression detection probe (20×, CYP3A4, FAM-labeled), 1 μL per reaction; Taqman gene expression detection probe (20×, ACTB, VIC-labeled): 1 μL per reaction; nuclease-free water, 4 μL per reaction; final PCR cocktail volume, 16 μL.

[0391] Dispense the PCR cocktail into the wells of a real-time PCR plate at room temperature. Dilute the cDNA samples 3-fold with nuclease-free water. Add 4 μL of the diluted cDNA sample to each PCR cocktail aliquot to a final volume of 20 μL, cover the plate, and mix gently. Then, briefly centrifuge to collect the contents at the bottom of the wells. In two wells of each PCR plate, add 4 μL of the RT mixture without cell lysate to the PCR cocktail as a negative control. The standard curve template is prepared from a 3-fold serial dilution of the cDNA sample mixture of the corresponding rifampicin-induced sample, with the highest concentration used.

[0392] Place the reaction into the QPCR system and start the run using the real-time PCR cycling program.

[0393] 4) Data Analysis: mRNA Level Determination

[0394] All calculations were performed using Microsoft Excel. The mRNA content in each vial was expressed as 2Ct(ACTB)-Ct(CYP). The mRNA level was determined by the following formula: Induction fold = mRNA (induced) / mRNA (solvent control). The results showed that the compounds in this application had no significant CYP induction activity.

[0395] Experimental Example 13: Caco-2 permeability test

[0396] 1) Cell culture:

[0397] Caco-2 cells were cultured in DMEM (DMEM) at 37°C, 5% CO₂. DMEM contained 10% fetal bovine serum, penicillin-streptomycin solution (100 U / mL and 0.1 mg / mL), 1% MEM non-essential amino acids, and 25 μM HEPES. Cells were seeded in 24-well Transwell plates at a density of 1.00 × 10 5 Cells were cultured in a CO2 incubator for 21 days before use in transport experiments, during which the culture medium was replaced every three days.

[0398] 2) Solution preparation:

[0399] The dosing solutions were roxithromycin solution, a high efflux reference substance, metoprolol solution, a low osmotic reference substance, atenolol solution, and the test compound solution. HBSS was used to prepare the dosing and receiving solutions.

[0400] Solution preparation for the drug transport experiment from apical to basal (AB): the 10 μM dosing solution at the A end contained 5 μM fluorescein and 0.4% DMSO; the receiving solution at the B end contained 0.4% DMSO.

[0401] Solution preparation for the basal to apical drug transport experiment (BA): the receiving solution at the A end contained 5 uM fluorescein and 0.4% DMSO; the dosing solution at the B end contained 10 μM 0.4% DMSO.

[0402] 3) Incubation step:

[0403] ①Quality control test before permeability test:

[0404] Before the experiment, the cell transmembrane resistance was measured with a resistance meter and the apparent transmembrane resistance of the single-layer cell membrane was calculated.

[0405] ②Permeability test:

[0406] Before the experiment, the cell culture medium in the culture plate was aspirated and then HBSS buffer pre-warmed at 37°C was added to wash three times (Apical end and Basal end).

[0407] Drug AB transport experiment: Aspirate the buffer in the plate, add 800 μL of 37°C pre-warmed HBSS buffer to the B end, add 600 μL of 37°C pre-warmed roxithromycin solution, metoprolol solution, atenolol solution and test compound solution to the A end respectively, and remove 100 μL of solution at the A end as the 0-minute A end sample and store at -20°C until testing.

[0408] Drug BA transport experiment: aspirate the buffer in the plate, add 500 μL of 37°C pre-warmed HBSS buffer to end A, add 900 μL of 37°C pre-warmed roxithromycin solution, metoprolol solution, atenolol solution and test compound solution to end B respectively, and remove 100 μL of solution at end B as the 0-minute end B sample and store at -20°C until testing.

[0409] Place the culture plate in a cell culture incubator and incubate for 90 minutes.

[0410] After the incubation, 100 μL of solution was taken from the A end and the B end of all samples as 90-minute samples and stored at -20°C until testing.

[0411] Standard curves were prepared for roxithromycin, metoprolol, atenolol, and the analytes. All samples were mixed with acetonitrile containing the internal standard and analyzed by LC-MS / MS.

[0412] ③Permeability test quality control test (check the integrity of cell membrane):

[0413] At 0 minutes, 100 μL of solution was removed from the A end of each AB and BA and transferred to a black 96-well plate. After 90 minutes of incubation, 100 μL of solution was removed from the B end of each AB and BA and transferred to a black 96-well plate. Fluorescence intensity was measured using a fluorescence microplate reader at an excitation wavelength of 485 nm and an emission wavelength of 527 nm.

[0414] 4) Data Analysis

[0415] Drug permeability: Apparent permeability coefficient (P app ) = (volume of solution at the receiving end / (membrane surface area × time)) × (drug concentration at the receiving end at 90 minutes × dilution factor) / (drug concentration at the administration end at 0 minutes × dilution factor)

[0416] where time is the total transit time in seconds.

[0417] Recovery rate: Recovery rate (%) = 100 × (drug concentration at the receiving end at 90 minutes × volume of solution at the receiving end × dilution factor + drug concentration at the administration end at 90 minutes × volume of solution at the administration end × dilution factor) / (drug concentration at the administration end at 0 minutes × volume of solution at the administration end × dilution factor).

[0418] Efflux ratio (ER) = apparent permeability coefficient from BA direction (P app ) / from the apparent permeability coefficient of AB (P app )

[0419] 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, isotopically labeled compound or prodrug thereof, wherein the compound has the structure of formula (I): Wherein: represents a single bond or a double bond, provided that two double bonds are not directly connected; W 1 , W 2 , W 3 and W 4 Each independently is C or N, provided that C is connected to a double bond; preferably, W 1 and W 2 At least one of them is N, and / or W 3 and W 4 At least one of is N; Selected from R 1 , R 3 , R 21 and R 22 is independently selected at each occurrence from H, a deuterium atom, a halogen, -OH, -NH2, -CN, -NO2, -SF5, =CH2, 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 、-S(=O)(=NR 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; When m is greater than 1, two Rs located on the same ring atom or adjacent ring atoms 3 together with the groups they are attached to optionally jointly form a C 3-6 hydrocarbon ring, 3- to 10-membered heterocyclic ring, C 6-10 aromatic ring or 5- to 14-membered heteroaromatic ring; R 4 For 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 41 selected from C 3-6 hydrocarbon ring, 3- to 10-membered heterocyclic ring, C 6-10 aryl ring, and 5- to 14-membered heteroaryl ring; R, R a and R b each independently selected from H, C 1-6 alkyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclic group, C 6-10 aryl, 5- to 14-membered heteroaryl and C 6-12 aralkyl; Ring B, ring X and ring Z are each independently selected from C 3-6 hydrocarbon rings, 3- to 10-membered heterocycles, C 6-10 aromatic rings and 5- to 14-membered heteroaromatic rings; Ring Y is absent or selected from C 3-6 hydrocarbon ring, 3- to 10-membered heterocyclic ring, C 6-10 aromatic ring and 5- to 14-membered heteroaromatic ring; when ring Y is absent, R 22 is also absent; The above alkylene, alkyl, alkenyl, alkynyl, cycloalkylene, cycloalkyl, hydrocarbon ring, heterocyclic group, heterocyclic ring, aryl, aromatic ring, heteroaryl, heteroaromatic ring and aralkyl are each optionally substituted with one or more substituents independently selected from the following: deuterium atoms, halogen, -OH, =O, -NH2, -CN, -NO2, =CH2, C 1-6 Alkyl, deuterated 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 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 , wherein the alkylene, alkyl, alkenyl, =CH2, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and aralkyl are each 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 Arylalkyl, -C 1-6 Alkylene-C 3-6 Cycloalkyl, -O-C 1-6 Alkyl and -C 1-6 Alkylene-O-C 1-6 Alkyl; R c and R d each independently selected from H, C 1-6 alkyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclic group, C 6-10 aryl, 5- to 14-membered heteroaryl, and C 6-12 aralkyl each time it appears, and the alkyl, cycloalkyl, heterocyclic group, 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- to 10-membered heterocyclic group, C 6-10 aryl, 5- to 14-membered heteroaryl, C 6- 12 aralkyl, and -C 1-6 alkylene-O-C 1-6 alkyl; and p, q and m are each independently an integer selected from 1, 2 or 3.

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 a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein: Ring B is C 3-6 a hydrocarbon ring or a 3- to 10-membered heterocyclic ring; Preferably, ring B is a cyclopentene ring, a cyclohexene ring, a pyrrolidine ring, an oxazolidine ring, a piperidine ring, a morpholine ring or an azepane ring.

4. The compound of any one of claims 1-3 or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein: R 1 Each occurrence is independently selected from H, halogen, C 1-6 alkyl, C 3-6 cycloalkyl, 3- to 10-membered heterocyclic group, C 6-10 aryl, 5- to 14-membered heteroaryl, -S(=O)2R a , -OR a and -NR a R b ; where each occurrence of R a and R b is independently selected from H, C 1-6 alkyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclic group, C 6-10 aryl, 5- to 14-membered heteroaryl and C 6-12 aralkyl; preferably, each occurrence of R 1 is independently selected from 3- to 10-membered heterocyclic group, C 6-10 aryl, 5- to 14-membered heteroaryl and -NR a R b ; provided that the alkyl, cycloalkyl, heterocyclic group, aryl and heteroaryl are each optionally substituted with one or more substituents independently selected from: halogen, -S(=O)2R c , C 1-6 alkyl, C 2-6 alkenyl, =CH2, C 3-6 cycloalkyl, 3- to 10-membered heterocyclic group, C 6-10 aryl and 5- to 14-membered heteroaryl; the alkyl, alkenyl, =CH2, cycloalkyl, heterocyclic group, aryl and heteroaryl are each further optionally substituted with one or more substituents independently selected from: halogen, C 1-6 alkyl, C 3-6 cycloalkyl, 3- to 10-membered heterocyclic group and -C 1-6 alkylene-C 3-6 cycloalkyl; Preferably, R 1 is independently C 6-10 aryl, -NR a R b ; Preferably, R 1 is H, methyl, halogen, methoxy, More preferably Most preferably, R 1 is 5. The compound of any one of claims 1-4 or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein ring X is a benzene ring, a 5- or 6-membered heterocycle or a 5- or 6-membered heteroaromatic ring and ring Y does not exist; or ring X is a benzene ring and ring Y is a 5- or 6-membered heterocycle or a 5- or 6-membered heteroaromatic ring; Preferably, For More preferably 6. A compound according to any one of claims 1 - 5 or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein R 21 and R 22 each independently, upon each occurrence, is selected from H, halogen, -SF5, C 1-6 alkyl, C 3-6 cycloalkyl, 3 - 10 - membered heterocyclic group, -O-(C 1-6 alkyl), -S(=O)2-(C 1-6 alkyl), -S(=O)2-(C 3-6 cycloalkyl), -S(=O)(=NR a )R b , -P(=O)(C 1-6 alkyl)2, (-C 3-6 cycloalkylidene)-CN and (-C 3-6 cycloalkylidene)-C 1-6 alkyl, and the alkyl, cycloalkylidene, cycloalkyl and heterocyclic group are each optionally substituted by one or more substituents independently selected from: halogen, C 1-6 alkyl and halo - C 1-6 alkyl; Preferably, R 21 and R 22 are each independently selected from H, halogen, -SF5, C 1-6 alkyl, C 3-6 cycloalkyl, 3- to 10-membered heterocyclic group, and -O-(C 1-6 alkyl), where the alkyl, cycloalkyl, and heterocyclic group are each optionally substituted with one or more substituents independently selected from: halogen, C 1-6 alkyl, and halo C 1-6 alkyl.

7. A compound according to any one of claims 1-6 or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein selected from:

8. A compound according to any one of claims 1-7 or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein R 3 is H, C 1-6 alkyl, -OR a or -SR a ; preferably, R 3 is H or C 1-6 alkyl; More preferably, R 3 is H, methyl, ethyl, -O-CH3 or -S-CH3; most preferably, R 3 is H or methyl; When m is greater than 1, two Rs located on the same ring atom or adjacent ring atoms 3 together with the groups to which they are attached optionally jointly form a C 3-6 hydrocarbon ring (preferably a cyclopropyl ring), and the hydrocarbon ring is optionally substituted with one or more substituents independently selected from the following: halogen, C 1-6 alkyl and halo C 1-6 alkyl.

9. The compound of any one of claims 1-8 or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein ring Z is a 3- to 10-membered heterocycle or a benzene ring; preferably a 5- to 10-membered heterocycle; more preferably a 5- or 6-membered heterocycle; and The heterocyclic ring and the benzene ring are each optionally substituted, each time they appear, by one or more substituents independently selected from the following: halogen, C 1-6 alkyl and halo-C 1-6 alkyl; Preferably, the ring Z is 10. A compound according to any one of claims 1-9, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein L 2 is -C(=O)-, C 1-6 alkylene or -NRC(=O)-, wherein R is H or C 1-6 alkyl; Preferably, L 2 is -C(=O)-, -CH2-, -CD2-.

11. A compound according to any one of claims 1-10 or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein R 41 is selected from 3- to 10-membered heterocycles, C 6-10 aromatic rings and 5- to 14-membered heteroaromatic rings, each of said heterocycles, aromatic rings and heteroaromatic rings being optionally substituted by one or more substituents independently selected from the following: halogen, -OH, C 1-6 alkyl, -O-C 1-6 alkyl and -S-C 1-6 alkyl, preferably, said heterocycles, aromatic rings and heteroaromatic rings are at least substituted by -OH or -O-C 1-6 alkyl; Preferably, -L 2 -R 41 is 12. The compound according to any one of claims 1-11, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein R 41 is a 5- to 6-membered heteroaryl ring, preferably a 6-membered heteroaryl ring, more preferably a pyridine ring or a pyrimidine ring, which is substituted by at least one -OH; Preferably, -L 2 -R 41 is 13. A compound according to any one of claims 1-12, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein the compound is selected from:

14. A pharmaceutical composition comprising a prophylactically or therapeutically effective amount of the compound of any one of claims 1-13 or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, and a pharmaceutically acceptable carrier.

15. Use of the compound of any one of claims 1-13 or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof or the pharmaceutical composition of claim 14 in the preparation of a drug for use as a WRN inhibitor, preferably, the drug is for preventing or treating 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 cancer, cervical cancer, esophageal cancer, breast cancer, renal cancer, prostate cancer and ovarian cancer.

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