WO2024255790A1 - Fused ring compounds, pharmaceutical composition comprising same and use thereof - Google Patents

Abstract

Disclosed are compounds of formula (I) useful as Werner helicase (WRN) inhibitors, a pharmaceutical composition comprising same, and the use thereof in the prevention or treatment of cancers characterized by microsatellite instability-high (MSI-H) or defective mismatch repair (dMMR). <img file="PCTCN2024098964-isre-I000001.jpg" he="68.79" img-content="drawing" img-format="jpg" inline="yes" orientation="portrait" wi="92.86"/>

Description

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

The present invention relates to a condensed ring compound, a pharmaceutical composition containing the condensed ring compound, and use thereof for preventing or treating diseases.

Background of the Invention

Microsatellite instability (MSI) is a type of genomic damage caused by mismatch repair deficiency (dMMR). Although progress has been made in the treatment of 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, among patients with dMMR colorectal cancer treated with PD-1 and PD-L1 checkpoint inhibitors, approximately half of patients experience primary drug resistance. See, e.g., Overman MJ et al., J Clin Oncol. 2018 Mar 10;36(8):773-779. There remains an unmet clinical need for new treatment options for patients who are refractory to currently available therapies.

Werner helicase (WRN) is considered a synthetic lethal target for MSI-H cancers (see, e.g., Chan EM et al., Nature. 2019 Apr; 568(7753): 551-556). WRN is a member of the RecQ family of DNA helicases and plays an important role in maintaining genome stability, DNA repair, replication, transcription, and telomere maintenance. Studies on WRN-dependent mechanisms have shown that dinucleotide TA repeats are massively amplified in MSI cells. These amplified TA repeats form secondary DNA structures that require WRN helicases to unwind (see, e.g., van Wietmarschen N et al., Nature. 2020 Oct; 586(7828): 292-298). In the absence of WRN (or when WRN helicases are inhibited), amplified TA repeats in MSI cells are cleaved by nucleases, ultimately leading to chromosome breaks. Therefore, inhibition of WRN helicase is an effective strategy for the treatment of cancers characterized by microsatellite instability-high (MSI-H) or mismatch repair deficiency (dMMR), including colorectal, gastric, or endometrial cancer.

SUMMARY OF THE INVENTION

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 less side effects), less prone to drug resistance.

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 a structure of formula (I):

in:

Selected from Where R ¹ is at the position marked with "*" in the above group connected, R ² is connected to the position marked with "**" in the above group, R ³ is connected to the position marked with "***" in the above group, and R ⁴ is connected to the position marked with "****" in the above group;

R ¹ is selected from -NR ¹¹ R ¹² ,

^R2 is

^R4 is

R ³ , R ¹¹ , R ¹² , R ¹³ , R ²¹ , R ²² , R ²³ and R ²⁴ are each independently selected at each occurrence from H, halogen, -OH, -NH ₂ , -CN, -NO ₂ , -SF ₅ , 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 heterocyclyl, 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) ₂ R ^a , -S(═O) ₂ NR ^a R ^{b -NRaRb} , -C(=O) ^NRaRb , -NRa ^- C(=O) ^Rb , -NRa ^- C(=O) ^ORb , -NRa ^- S(=O) ₂ - ^Rb , ^{-NRa - C} (=O ⁾ -NRaRb, _-P ⁽ ₌ O ⁾ _RaRb , -C1-6alkylene _- ^Ra , -C1-6alkylene-ORa ^, _{-C1-6alkylene} - ^NRaRb , -OC1-6alkylene-NRaRb, ( _{-C3-6cycloalkylene} ) ^{-CN and} ( ^{-C3-6cycloalkylene} ) ^{-C1-6haloalkyl} _;

R ¹⁴ at each occurrence is independently selected from -L ¹ -(C _3-6 cycloalkyl), -L ¹ -(3-10 membered heterocyclyl), -L ¹ -(C _6-10 aryl) and -L ¹ -(5-14 membered heteroaryl);

R ⁴¹ is selected from a C _3-6 hydrocarbon ring, a 3-10 membered heterocyclic ring, a C _6-10 aromatic ring, and a 5-14 membered heteroaromatic ring;

Ring C, Ring D, Ring E, Ring X and Ring Z are each independently selected from a C _3-6 hydrocarbon ring, a 3-10 membered heterocyclic ring, a C _6-10 aromatic ring and a 5-14 membered heteroaromatic ring;

Ring Y is absent or is selected from a C _3-6 hydrocarbon ring, a 3-10 membered heterocyclic ring, a C _6-10 aromatic ring and a 5-14 membered heteroaromatic ring; when Ring Y is absent, R ²⁴ is also absent;

L ¹ is selected from a direct bond, -O-, -C(=O)-, -NRC(=O)-, -S-, -S(=O)-, -S(=O) ₂ -, C _1-6 alkylene, and -O-(C _1-6 alkylene)-;

^L2 is selected from -O-, -C(=O)-, -NRC(=O)-, -S-, -S(=O)-, -S(=O) _2- , _C1-6 alkylene and -O-( _C1-6 alkylene)-;

R, ^Ra and ^Rb are each independently selected at each occurrence from H, _C1-6 alkyl, _C3-10 cycloalkyl, 3-10 membered heterocyclyl, _C6-10 aryl, 5-14 membered heteroaryl and _C6-12 aralkyl;

The above alkylene, alkyl, alkenyl, alkynyl, cycloalkylene, cycloalkyl, hydrocarbon ring, heterocyclyl, heterocycle, aryl, aromatic ring, heteroaryl, heteroaromatic ring and aralkyl are each optionally substituted at each occurrence by one or more substituents independently selected from the following: halogen, -OH, =O, = _CF2 , _-NH2 , -CN, _-NO2 , _C1-6 alkyl, deuterated _C1-6 alkyl, C3-6 cycloalkyl, _3-10 membered heterocyclyl, _C6-10 aryl, 5-14 membered heteroaryl, _C6-12 aralkyl, -C(=O) ^Rc , -OC(=O) ^Rc , -C(=O) ^ORc , ^-ORc , -SRc, -S(=O ^{)Rc ,} -S(= ^O ) _2Rc ^, -S(=O ⁾ _2NRcRd ^{, -NRcRd} -C(═O ^{) NRcRd} , -NRc- ^C (═O) ^Rd , -NRc- ^C (═O) ^ORd , -NRc ^- S(═O) ₂ - ^Rd , -NRc ^{-C (} ═O) ^-NRcRd , _{-C1-6alkylene} - ^ORc , _{-C1-6alkylene} - ^NRcRd and -OC1-6alkylene- ^NRcRd , said alkylene, alkyl, cycloalkyl, heterocyclyl, aryl, _heteroaryl and aralkyl being 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 , _C1-6alkyl , _{C1-6haloalkyl} , C3-6cycloalkyl, _3-10 membered heterocyclyl, C _6-10- membered aryl, 5-14-membered heteroaryl, C _6-12 aralkyl, -OC _1-6 alkyl, and -C _1-6 alkylene-OC _1-6 alkyl;

R ^c and R ^d are each independently selected at each occurrence from H, C _1-6 alkyl, C _3-10 cycloalkyl, 3-10 membered heterocyclyl, C _6-10 aryl, 5-14 membered heteroaryl and C _6-12 aralkyl, said alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and aralkyl further optionally substituted with one or more substituents independently selected from halogen, -OH, =O, -C(=O)O-tert-butyl, -NH ₂ , -CN, -NO ₂ , C _1-6 alkyl, C _1-6 haloalkyl, C _3-6 cycloalkyl, 3-10 membered heterocyclyl _, C _6-10 aryl, 5-14 membered heteroaryl, C _6-12 aralkyl and -C _1-6 alkylene-OC _1-6 alkyl;

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

m is an integer of 0 or 1.

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.

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.

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.

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 an individual 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.

Detailed description of the invention

definition

Unless otherwise defined below, the meanings of all technical terms and scientific terms used herein are intended to be the same as those generally understood by those skilled in the art. Reference to the technology used herein is intended to refer to the technology generally understood in the art, including those changes in technology or replacement of equivalent technology that are obvious to those skilled in the art. Although it is believed that the following terms are well understood by those skilled in the art, the following definitions are still set forth to better explain the present invention.

"Zwitterion" or "zwitterionic form" means a compound containing both positively charged and negatively charged functional groups. In some examples, certain groups are in non-zwitterionic form per se, but in a tautomer in which a proton is shifted, the group is in zwitterionic form. For example, when the R ⁴¹ group in the compound described herein is a pyridine ring substituted with at least one -OH, it may be in the following zwitterionic form (c) or non-zwitterionic form (d) or a mixture of the two forms:

wherein R ₁₀₀ , R ₁₀₁ , and R ₁₀₂ represent substituents as defined herein;

For another example, when the R ⁴¹ group in the compound described herein is a pyrimidine ring substituted with at least one -OH, it may be in the following zwitterionic form (a) or (b) or non-zwitterionic form (e), or a mixture of any two or three of the forms:

wherein R ₁₀₀ and R ₁₀₂ represent substituents as defined herein.

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.

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.

As used herein, the term "alkyl" is defined as a straight or branched saturated aliphatic hydrocarbon. In some embodiments, the alkyl has 1 to 12, for example 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 by 1 or more (such as 1 to 3) suitable substituents such as halogen (in this case, the group is referred to as "haloalkyl") (e.g., CF ₃ , C ₂ F ₅ , CHF ₂ , CH ₂ F, CH ₂ CF ₃ , CH ₂ Cl or -CH ₂ CH ₂ CF _3, 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).

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 (" _C2-6 alkenyl"). 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 compounds of the present invention contain an alkenyl group, the compounds may be present 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 alkenylene", etc., and specific examples include, but are not limited to: -CH=CH-, -CH ₂ CH=CH-, -C(CH ₃ )=CH-, butenylene, pentenylene, hexenylene, etc.

As used herein, the term "alkynyl" refers to a monovalent hydrocarbon group 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 group, including, for example, "C _2-8 alkynylene", "C _2-6 alkynylene", "C _2-4 alkynylene", etc. Examples thereof 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.

As used herein, the term "paracyclic ring" or "fused ring" refers to a ring system formed by two or more cyclic structures sharing two adjacent atoms with each other.

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.

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.

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 (ring), (cyclo)butyl (ring), (cyclo)pentyl (ring), (cyclo)hexyl (ring), (cyclo)heptyl (ring), (cyclo)octyl (ring), (cyclo)nonyl (ring), (cyclo)hexenyl (ring) and the like.

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 a spirocyclic, fused or bridged system (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 1 or more (such as 1 to 3) suitable substituents. The cycloalkyl group has 3 to 15 carbon atoms. For example, the term " _{C3-6cycloalkyl} " refers to a saturated monocyclic or polycyclic (such as bicyclic) hydrocarbon ring of 3 to 6 ring carbon atoms (e.g., cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl), which is optionally substituted with 1 or more (such as 1 to 3) suitable substituents, for example, methyl-substituted cyclopropyl.

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 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 oxirane, 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.

As used herein, the term "heterocyclyl" (or "heterocycle") encompasses fused ring structures, where the point of attachment to other groups can be on either ring of the fused ring structure. Therefore, the heterocyclic group of the present invention also includes, but is not limited to, heterocyclic and heterocyclic groups, heterocyclic and cycloalkyl groups, monoheterocyclic and monoheterocyclic groups, monoheterocyclic and monocycloalkyl groups, aryl and heterocyclic groups, heteroaryl and heterocyclic groups, such as 3-7 membered (mono) heterocyclic groups and 3-7 membered (mono) heterocyclic groups, 3-7 membered (mono) heterocyclic groups and (mono) cycloalkyl groups, 3-7 membered (mono) heterocyclic groups and C _4-6 (mono) cycloalkyl groups, C _6-10 aryl and 3-7 membered heterocyclic groups, 5-6 membered heteroaryl and 3-7 membered heterocyclic groups, 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,

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

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.

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 spiro heterocycle", "oxygen-containing spiro heterocycle" and "sulfur-containing spiro heterocycle" optionally further contain one or more other heteroatoms selected from oxygen, nitrogen and sulfur. The term "6-10 membered nitrogen-containing spiro heterocyclic group" refers to a spiro heterocyclic group containing a total of 6-10 ring atoms and at least one of the ring atoms being a nitrogen atom.

As used herein, the terms "(ylidene)aryl" 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 terms "C _6-10 (ylidene)aryl" and "C _6-10 aromatic ring" mean an aromatic group containing 6 to 10 carbon atoms, such as (ylidene)phenyl (benzene ring) or (ylidene)naphthyl (naphthalene ring). The (ylidene)aryl group and the aromatic ring are optionally substituted with 1 or more (such as 1 to 3) suitable substituents (e.g., halogen, -OH, -CN, -NO ₂ , C _1-6 alkyl, etc.).

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

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 additionally may be benzo-fused. In particular, "(ene)heteroaryl" or "heteroaryl ring" is selected from (ene)thienyl (ring), (ene)furanyl (ring), (ene)pyrrolyl (ring), (ene)oxazolyl (ring), (ene)thiazolyl (ring), (ene)imidazolyl (ring), (ene)pyrazolyl (ring), (ene)isoxazolyl (ring), (ene)isothiazolyl (ring), (ene)oxadiazolyl (ring), (ene)triazolyl (ring), (ene)thiadiazolyl (ring), etc., and their benzo derivatives; or (ene)pyridinyl (ring), (ene)pyridazinyl (ring), (ene)pyrimidinyl (ring), (ene)pyrazinyl (ring), (ene)triazinyl (ring), etc., and their benzo derivatives.

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

As used herein, the term "alkylthio" refers to an alkyl group as defined above attached to the parent molecular moiety through a sulfur atom. Representative examples of _{C 1-6} alkylthio include, but are not limited to, methylthio, ethylthio, tert-butylthio and hexylthio.

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 also optionally contain one or more (e.g., one, two, three or four) ring members selected from N, O, S, S=O and S(=O) ₂ ; 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.

The term "substituted" means that one or more (e.g., one, two, three, or four) hydrogens on the designated atom are replaced by a selection from the indicated group, provided that the normal valence of the designated atom in the present context is not exceeded and the substitution forms a stable compound. Combinations of substituents and/or variables are permitted only if such combinations form stable compounds.

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

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.

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

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

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.

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)); isotopes of carbon (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 (e.g., ¹²³ I and ¹²⁵ I); isotopes of nitrogen (e.g., ¹³ N and ¹⁵ N); isotopes of oxygen (e.g., ¹⁵ O, ¹⁷ O, and ¹⁸ O); isotopes of phosphorus (e.g., ³² P); and isotopes of sulfur (e.g., ³⁵ S). Certain isotopically labeled compounds of the invention (e.g., those incorporating radioisotopes) are useful in drug and/or substrate tissue distribution studies (e.g., assays). The radioisotopes tritium (i.e., ³ H) and carbon-14 (i.e., ¹⁴ C) are particularly useful for this purpose because they are easily incorporated and easily detected. Substitution with positron emitting isotopes (e.g., ¹¹ C, ¹⁸ F, ¹⁵ O, and ¹³ N) can be used to examine substrate receptor occupancy in positron emission tomography (PET) studies. Isotopically labeled compounds of the invention can be prepared by methods similar to those described in the accompanying routes and/or in the examples and preparations by using appropriate isotopically labeled reagents in place of the non-labeled reagents previously employed. Pharmaceutically acceptable solvates of the invention include those in which the crystallization solvent may be isotopically substituted, for example, D ₂ O, acetone-d ₆ or DMSO-d ₆ .

The term "stereoisomer" means 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, it can produce racemic mixtures, single enantiomers, diastereomeric mixtures, and individual diastereomers. Specific individual molecules can also exist as geometric isomers (cis/trans). Similarly, the compounds of the present invention can exist as mixtures (commonly referred to as tautomers) of two or more structurally different forms in rapid equilibrium. 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 the present application covers all such isomers or mixtures thereof in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%).

In this article, solid lines can be used Solid wedge Virtual wedge Carbon-carbon bonds of the compounds of the 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 stereoisomer shown is present. When present in a racemic mixture, the solid and dashed wedges are used to define relative stereochemistry, not absolute stereochemistry. Unless otherwise indicated, the compounds of the invention are intended to exist in the form of stereoisomers, which include 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 invention may exhibit more than one type of isomerism and consist of mixtures thereof (e.g., racemic mixtures and diastereomeric pairs).

Atropisomers are compounds that can be separated into their rotationally restricted isomers.

It should also be understood that certain compounds of the present invention may be present in free form for treatment, 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, which, upon administration to a patient in need thereof, are capable of directly or indirectly providing the compounds of the present invention. or its metabolites or residues. Therefore, when referring to "compounds of the present invention" herein, the above-mentioned various derivative forms of the compounds are also intended to be encompassed.

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

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.

As used herein, the term "ester" means an ester 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 free acid or alcohol form). The compounds of the present invention themselves may also be esters.

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

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, etc. of the administered compound. Thus, the present invention includes metabolites of the compounds of the present invention, including compounds prepared by contacting the compounds of the present invention with a mammal for a period of time sufficient to produce a metabolic product thereof.

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 when administered into or onto the body, for example, by hydrolytic cleavage. Typically such prodrugs will be functional group derivatives of the compounds that are easily 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," Vol. 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)).

The present invention also encompasses compounds of the present invention containing protecting groups. In 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 by conventional protecting groups, for example, those described in Protective Groups in Organic Chemistry, ed. J.F.W. McOmie, Plenum Press, 1973; and T.W. Greene & P.G.M. Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991, which references are incorporated herein by reference. The protecting groups may be removed at an appropriate subsequent stage using methods known in the art.

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

Compound

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):

in:

Selected from wherein R ¹ is connected to the position marked with “*” in the above group, R ² is connected to the position marked with “**” in the above group, R ³ is connected to the position marked with “***” in the above group, and R ⁴ is connected to the position marked with “****” in the above group;

R ¹ is selected from -NR ¹¹ R ¹² ,

^R2 is

^R4 is

R ³ , R ¹¹ , R ¹² , R ¹³ , R ²¹ , R ²² , R ²³ and R ²⁴ are each independently selected at each occurrence from H, halogen, -OH, -NH ₂ , -CN, -NO ₂ , -SF ₅ , 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 heterocyclyl, 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) ₂ R ^a , -S(═O) ₂ NR ^a R ^{b -NRaRb} , -C(=O) ^NRaRb , -NRa ^- C(=O) ^Rb , -NRa ^- C(=O) ^ORb , -NRa ^- S(=O) ₂ - ^Rb , ^{-NRa - C} (=O ⁾ -NRaRb, _-P ⁽ ₌ O ⁾ _RaRb , -C1-6alkylene _- ^Ra , -C1-6alkylene-ORa ^, _{-C1-6alkylene} - ^NRaRb , -OC1-6alkylene-NRaRb, ( _{-C3-6cycloalkylene} ) ^{-CN and} ( ^{-C3-6cycloalkylene} ) ^{-C1-6haloalkyl} _;

R ¹⁴ at each occurrence is independently selected from -L ¹ -(C _3-6 cycloalkyl), -L ¹ -(3-10 membered heterocyclyl), -L ¹ -(C _6-10 aryl) and -L ¹ -(5-14 membered heteroaryl);

R ⁴¹ is selected from a C _3-6 hydrocarbon ring, a 3-10 membered heterocyclic ring, a C _6-10 aromatic ring, and a 5-14 membered heteroaromatic ring;

Ring C, Ring D, Ring E, Ring X and Ring Z are each independently selected from a C _3-6 hydrocarbon ring, a 3-10 membered heterocyclic ring, a C _6-10 aromatic ring and a 5-14 membered heteroaromatic ring;

Ring Y is absent or is selected from a C _3-6 hydrocarbon ring, a 3-10 membered heterocyclic ring, a C _6-10 aromatic ring and a 5-14 membered heteroaromatic ring; when Ring Y is absent, R ²⁴ is also absent;

L ¹ is selected from a direct bond, -O-, -C(=O)-, -NRC(=O)-, -S-, -S(=O)-, -S(=O) ₂ -, C _1-6 alkylene, and -O-(C _1-6 alkylene)-;

^L2 is selected from -O-, -C(=O)-, -NRC(=O)-, -S-, -S(=O)-, -S(=O) _2- , _C1-6 alkylene and -O-( _C1-6 alkylene)-;

R, ^Ra and ^Rb are each independently selected at each occurrence from H, _C1-6 alkyl, _C3-10 cycloalkyl, 3-10 membered heterocyclyl, _C6-10 aryl, 5-14 membered heteroaryl and _C6-12 aralkyl;

The above alkylene, alkyl, alkenyl, alkynyl, cycloalkylene, cycloalkyl, hydrocarbon ring, heterocyclyl, heterocycle, aryl, aromatic ring, heteroaryl, heteroaromatic ring and aralkyl are each optionally substituted at each occurrence by one or more substituents independently selected from the following: halogen, -OH, =O, = _CF2 , _-NH2 , -CN, _-NO2 , _C1-6 alkyl, deuterated _C1-6 alkyl, C3-6 cycloalkyl, _3-10 membered heterocyclyl, _C6-10 aryl, 5-14 membered heteroaryl, _C6-12 aralkyl, -C(=O) ^Rc , -OC(=O) ^Rc , -C(=O) ^ORc , ^-ORc , -SRc, -S(=O ^{)Rc ,} -S(= ^O ) _2Rc ^, -S(=O ⁾ _2NRcRd ^{, -NRcRd} -C(═O ^{) NRcRd} , -NRc- ^C (═O) ^Rd , -NRc- ^C (═O) ^ORd , -NRc ^- S(═O) ₂ - ^Rd , -NRc ^{-C (} ═O) ^-NRcRd , _{-C1-6alkylene} - ^ORc , _{-C1-6alkylene} - ^NRcRd and -OC1-6alkylene- ^NRcRd , said alkylene, alkyl, cycloalkyl, heterocyclyl, aryl, _heteroaryl and aralkyl being 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 , _C1-6alkyl , _{C1-6haloalkyl} , C3-6cycloalkyl, _3-10 membered heterocyclyl, C _6-10 membered aryl, 5-14 membered heteroaryl, C _6-12 aralkyl, -OC _1-6 alkyl and -C _1-6 alkylene-OC _1-6 alkyl; preferably, the above alkylene, alkyl, alkenyl, alkynyl, cycloalkylene, cycloalkyl, hydrocarbon The ring, heterocyclyl, heterocycle, aryl, aromatic ring, heteroaryl, heteroaromatic ring and aralkyl are each optionally substituted at each occurrence by one or more substituents independently selected from the group consisting of halogen, -OH, = _O , _-NH2 , -CN, -NO2, Ci _-6 alkyl, deuterated Ci _-6 alkyl, C3-6 cycloalkyl, _3-10 membered heterocyclyl, _C6-10 aryl, 5-14 membered heteroaryl, _C6-12 aralkyl, -C(=O) ^Rc , -OC(=O) ^Rc , -C(=O) ^ORc , ^-ORc , ^-SRc , -S(=O) ^Rc _, -S(=O) _2Rc , -S(= ^O ) ^2NRcRd , ^-NRcRd , -C(= ^O ) ^NRcRd , ^{-NRc- C} (=O) ^{Rd ,} -NR wherein the ^alkylene , ^alkyl , cycloalkyl ^, heterocyclyl ^{, aryl} , ^heteroaryl and ^aralkyl are ^further optionally substituted with _{one or} more substituents independently selected from the group consisting of halogen, ^-OH , = ^O , ^-C (=O ⁾ O-tert-butyl, _{-NH2, -CN, -NO2, C1-6 alkyl, C1-6 haloalkyl , C3-6 cycloalkyl , 3-10} membered heterocyclyl, _C6-10 aryl, _5-14 membered heteroaryl, _C6-12 aralkyl, _-OC1-6 alkyl and _-C1-6 alkylene-OC _1-6 alkyl;

R ^c and R ^d are each independently selected at each occurrence from H, C _1-6 alkyl, C _3-10 cycloalkyl, 3-10 membered heterocyclyl, C _6-10 aryl, 5-14 membered heteroaryl and C _6-12 aralkyl, said alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and aralkyl further optionally substituted with one or more substituents independently selected from halogen, -OH, =O, -C(=O)O-tert-butyl, -NH ₂ , -CN, -NO ₂ , C _1-6 alkyl, C _1-6 haloalkyl, C _3-6 cycloalkyl, 3-10 membered heterocyclyl _, C _6-10 aryl, 5-14 membered heteroaryl, C _6-12 aralkyl and -C _1-6 alkylene-OC _1-6 alkyl;

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

m is an integer of 0 or 1.

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 a structure of the following formula:

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

^R1 is

R ¹³ is H, halogen or C _1-6 alkyl;

R ¹⁴ is selected from 3-10 membered heterocyclyl, -(C _1-6 alkylene)-(3-10 membered heterocyclyl) and -O-(C _1-6 alkylene)-(3-10 membered heterocyclyl), said heterocyclyl being optionally substituted with one or more substituents independently selected from the group consisting of C _1-6 alkyl, C _3-6 cycloalkyl, 3-10 membered heterocyclyl, -C _1-6 alkylene-C _3-6 cycloalkyl and -C _1-6 alkylene-CN; and

n is an integer of 1 or 2.

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

^R1 is

R ¹³ is H, halogen or C _1-6 alkyl;

R ¹⁴ is selected from 3-10 membered heterocyclyl, -(C _1-6 alkylene)-(3-10 membered heterocyclyl) and -O-(C _1-6 alkylene)-(3-10 membered heterocyclyl), wherein the heterocyclyl is optionally substituted with one or more substituents independently selected from the group consisting of C _1-6 alkyl, C _3-6 cycloalkyl, -C _1-6 alkylene-C _3-6 cycloalkyl and -C _1-6 alkylene-CN; and

n is an integer of 1 or 2.

In a preferred embodiment, in the general formula of the present disclosure: R ¹ is

In a preferred embodiment, in the general formula of the present disclosure: R ¹ is

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

^R1 is

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

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

The above-mentioned benzene ring, heterocycle and heteroaromatic ring are each optionally substituted by one or more substituents independently selected from the following: halogen, -OH, _-NH2 , -CN, _-NO2 , _C1-6 alkyl, deuterated _C1-6 alkyl, _C1-6 haloalkyl, _-OC1-6 alkyl, _C3-6 cycloalkyl and 3-10 membered heterocyclyl; preferably, the above-mentioned benzene ring, heterocycle and heteroaromatic ring are each optionally substituted by one or more substituents independently selected from the following: halogen, _C1-6 alkyl, _C1-6 haloalkyl, _-OC1-6 alkyl and _C3-6 cycloalkyl.

In a preferred embodiment, in the general formula of the present disclosure: R ¹ is

In a preferred embodiment, in the general formula of the present disclosure: R ¹ is

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

^R1 is ^{-NR11R12 ;}

R ¹¹ and R ¹² are each independently selected from H, C _1-6 alkyl, C _3-6 cycloalkyl, 3-10 membered heterocyclyl, C _6-10 aryl, 5-14 membered heteroaryl, C _6-12 aralkyl, -C _1-6 alkylene-R ^a and -C _1-6 alkylene-NR ^a R ^b ;

^Ra and ^Rb are each independently selected at each occurrence from H, _C1-6 alkyl, _C3-10 cycloalkyl, 3-10 membered heterocyclyl, _C6-10 aryl, 5-14 membered heteroaryl and _C6-12 aralkyl;

The alkylene, alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and aralkyl groups are each optionally substituted at each occurrence with one or more substituents independently selected from the group consisting of halogen and C _1-6 alkyl.

In a preferred embodiment, in the general formula of the present disclosure: R ¹ is

In a preferred embodiment, in the general formula of the present disclosure: R ¹ is

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

^R1 is

Ring E is a C _3-6 hydrocarbon ring, a 3-10 membered heterocyclic ring, a C _6-10 aromatic ring or a 5-14 membered heteroaromatic ring, wherein the hydrocarbon ring and the heterocyclic ring are optionally substituted by =CF ₂ ;

R ¹³ is H, halogen, C _1-6 alkyl, C _3-6 cycloalkyl, 3-10 membered heterocyclyl, -C(=O)-(C _1-6 alkyl), -O-(C _1-6 alkyl) or -N(C _1-6 alkyl) ₂ , wherein the alkyl, cycloalkyl and heterocyclyl are further optionally substituted by C _1-6 alkyl, C _3-6 cycloalkyl or 3-10 membered heterocyclyl;

n is an integer of 1 or 2.

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

^R1 is

Ring E is a C _3-6 hydrocarbon ring, a 3-10 membered heterocyclic ring, a C _6-10 aromatic ring or a 5-14 membered heteroaromatic ring;

R ¹³ is H, halogen, C _1-6 alkyl, 3-10 membered heterocyclyl, -C(═O)-(C _1-6 alkyl), -O-(C _1-6 alkyl) or -N(C _1-6 alkyl) ₂ , wherein the alkyl and heterocyclyl are further optionally substituted by C _1-6 alkyl or 3-10 membered heterocyclyl;

n is an integer of 1 or 2.

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

^R1 is

Ring E is a 3-10 membered heterocyclic ring, a C _6-10 aromatic ring or a 5-14 membered heteroaromatic ring;

R ¹³ is H, halogen, C _1-6 alkyl, -C(═O)-(C _1-6 alkyl) or -O-(C _1-6 alkyl);

n is an integer of 1 or 2.

In a preferred embodiment, in the general formula of the present disclosure: R ¹ is

In a preferred embodiment, in the general formula of the present disclosure: R ¹ is

In a preferred embodiment, in the general formula of the present disclosure: R ¹ is

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

When Y does not exist,

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

b). Ring X is a benzene ring, and at least one R ²³ is (—C _3-6 cycloalkylene)-CN or (—C _3-6 cycloalkylene)-C _1-6 haloalkyl.

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

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

In a preferred embodiment, in the general formula of the present disclosure: R ²¹ , R ²² , R ²³ and R ²⁴ are each independently selected from H, halogen, -SF ₅ , C _1-6 alkyl, halogenated C _1-6 alkyl, C _3-6 cycloalkyl, -S(＝O) ₂ -(C _1-6 alkyl), -S(＝O) ₂ -(C _3-6 cycloalkyl), -P(＝O)(C _1-6 alkyl) ₂ , (-C _3-6 cycloalkylene)-CN and (-C _3-6 cycloalkylene)-C _1-6 haloalkyl.

In a preferred embodiment, in the general formula of the present disclosure: R ² is selected from:

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

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

R ²³ at each occurrence is independently selected from H, halogen, -SF ₅ , C _1-6 alkyl, haloC _1-6 alkyl, C _3-6 cycloalkyl, -S(═O) ₂ -(C _1-6 alkyl), -S(═O) ₂ -(C _3-6 cycloalkyl), -P(═O)(C _1-6 alkyl) ₂ , (-C _3-6 cycloalkylene)-CN and (-C _3-6 cycloalkylene)-C _1-6 haloalkyl.

In a preferred embodiment, in the general formula of the present disclosure: R ² is selected from:

In a preferred embodiment, in the general formula of the present disclosure: R ³ is H or C _1-6 alkyl.

In a preferred embodiment, in the general formula of the present disclosure: R ³ is ethyl.

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

The heterocyclic ring and the benzene ring are each optionally substituted at each occurrence with one or more substituents independently selected from the group consisting of halogen and C _1-6 alkyl.

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

In a preferred embodiment, in the general formula of the present disclosure: L ² is -C(=O)- or -NRC(=O)-, wherein R is H or C _1-6 alkyl.

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

In a preferred embodiment, in the general formula of the present disclosure: R ⁴¹ is selected from a 3-10 membered heterocycle, a C _6-10 aromatic ring and a 5-14 membered heteroaromatic ring, and the heterocycle, aromatic ring and heteroaromatic ring are each optionally substituted by one or more substituents independently selected from the following: halogen, -OH, -CN, C _1-6 alkyl, -OC _1-6 alkyl and -SC _1-6 alkyl, preferably, the heterocycle, aromatic ring and heteroaromatic ring are at least substituted by -OH, -CN or -OC _1-6 alkyl.

In a preferred embodiment, in the general formula of the present disclosure: R ⁴¹ is selected from a 3-10 membered heterocycle, a C _6-10 aromatic ring and a 5-14 membered heteroaromatic ring, and the heterocycle, 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 heterocycle, aromatic ring and heteroaromatic ring are at least substituted by -OH or -OC _1-6 alkyl.

In a preferred embodiment, in the general formula of the present disclosure: -L ² -R ⁴¹ is

In a preferred embodiment, in the general formula of the present disclosure: -L ² -R ⁴¹ is

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

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

In a preferred embodiment, in the general formula of the present disclosure: -L ² -R ⁴¹ is

In a preferred embodiment, in the general formula of the present disclosure: -L ² -R ⁴¹ is

The present disclosure covers technical solutions/compounds obtained by any combination of various embodiments.

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:

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

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

Pharmaceutical compositions and methods of treatment

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 semisolid preparation, a liquid preparation or a gaseous preparation. In some embodiments, the pharmaceutical composition may also include one or more other therapeutic agents.

In some embodiments, the present invention provides the 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.

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.

In some embodiments, 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), comprising administering to an individual 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.

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.

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

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

As used herein, "individual" includes humans or non-human animals. Exemplary human individuals include human individuals (referred to as patients) suffering from diseases (e.g., diseases described herein) or normal individuals. "Non-human animals" in the present invention 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.).

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

General synthetic route:

Route 1

Route 2

Route 3

in:

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

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

M is a metal, a borate ester or a boric acid;

R ⁰ at each occurrence is independently C _1-6 alkyl.

In the above route, the synthesis route is described by taking the compound with unsubstituted piperazine ring as Z ring as an example. The above general synthesis route can also be used for the synthesis of compounds in which Z ring is substituted piperazine or fused piperazine analogs.

Example

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.

The abbreviations in this invention have the following meanings:

Example 1: Synthesis of Compound A1

1) Synthesis of intermediate A1-3:

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

2) Synthesis of intermediate A1-4:

2-Chloro-4-trifluoromethylaniline (10 g, 51.13 mmol) and potassium carbonate (14.13 g, 102.26 mmol) were dissolved in dichloromethane (100 mL) solution, bromoacetyl bromide (10.32 g, 51.13 mmol) was added dropwise under stirring at 0°C, and the mixture was stirred at 25°C for 1 hour. LCMS showed that the reaction was complete, and the reaction solution was poured into water, extracted with dichloromethane (250 mL*2), and concentrated to obtain a white solid A1-4 (12 g, yield: 74.2%). m/z[MH] ^- =314.0. ¹ H NMR (400MHz, DMSO-d ₆ ) δ10.21 (s, 1H), 8.10 (d, J = 8.5 Hz, 1H), 7.98 ( d, J = 2.1 Hz, 1H), 7.77 ( dd, J = 8.6, 2.1 Hz, 1H), 4.27 ( s, 2H).

3) Synthesis of intermediate A1-5:

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

4) Synthesis of intermediate A1-6:

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

5) Synthesis of product A1:

A1-7 (9.8 mg, 0.064 mmol), acetonitrile (1 mL), N-hydroxy-7-azabenzotriazole (8.7 mg, 0.064 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (15 mg, 0.08 mmol) were added to a 25 mL single-mouth bottle in sequence, and stirred at room temperature for 1 hour. A1-6 (35 mg, 0.053 mmol) and N,N-diisopropylethylamine (27 mg, 0.21 mmol) were added and stirred at room temperature for 1 hour. The reaction solution was filtered and purified by preparative chromatography (acetonitrile/water = 40%) to obtain product A1 (9 mg, yield: 21%). ESI (m/z) = 794.5 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.41(s,1H),8.58(s,1H),8.05(d,J＝8.6Hz,1H),7.99–7.90(m,3H),7.71(dd,J ＝8.8,2.1Hz,1H),7.03(d,J＝9.0Hz,2H),5.37(s,2H),4.53(d,J＝12.3Hz,1H),3.53 (d,J＝10.0Hz,4H),3.26(d,J＝4.8Hz,4H),2.99(d,J＝9.7Hz,3H),2.83(d,J＝11.0Hz ,1H),2.70–2.60(m,1H),2.46(d,J＝8.6Hz,7H),2.24(s,3H),1.20(t,J＝7.4Hz,3H).

Example 2: Synthesis of Compound A2

1) Synthesis of intermediate A2-1:

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

2) Synthesis of intermediate A2-2:

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

3) Synthesis of intermediate A2-3:

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

4) Synthesis of intermediate A2-4:

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

5) Synthesis of product A2:

A1-7 (11 mg, 0.07 mmol), acetonitrile (1 mL), N-hydroxy-7-azabenzotriazole (9.5 mg, 0.07 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (17 mg, 0.087 mmol) were added to a 25 mL single-mouth bottle in sequence, and stirred at room temperature for 1 hour. A2-4 (39 mg, 0.058 mmol) and N,N-diisopropylethylamine (30 mg, 0.23 mmol) were added and stirred at room temperature for 1 hour. The reaction solution was filtered and purified by preparative HPLC (acetonitrile/0.05% formic acid aqueous solution = 30%) to obtain product A2 (14 mg, yield: 30%), which was a formate salt. ESI (m/z) = 808.4 [M + H] ⁺ .1H NMR (400 MHz, DMSO-d ₆ )δ10.42(s,1H),8.56(s,1H),8.22(s,1H),8.08–8.03(m,3H),7.97(d,J ＝2.1Hz,1H),7.71(d,J＝9.1Hz,1H),7.43(d,J＝8.0Hz,2H),5.39(s,2H), 4.53(d,J＝12.6Hz,2H),3.52(d,J＝11.8Hz,13H),3.02(d,J＝9.3Hz,4H), 2.84(d,J＝11.4Hz,2H),2.44(s,3H),2.15(s,3H),1.20(d,J＝7.5Hz,3H).

Example 3: Synthesis of Compound A31

1) Synthesis of intermediate A31-2:

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

2) Synthesis of intermediate A31-3:

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

3) Synthesis of intermediate A31-4:

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

4) Synthesis of product A31:

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

Example 4: Synthesis of Compound A4

1) Synthesis of intermediate A4-2:

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

2) Synthesis of intermediate A4-3:

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

3) Synthesis of product A4:

Compound A4-3 (120 mg, 0.21 mmol), N-hydroxy-7-azabenzotriazole (31 mg, 0.23 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (60 mg, 0.32 mmol) were dissolved in acetonitrile (3 mL) solution and reacted for 1 hour under stirring at 25°C. Compound A4-4 (65 mg, 0.32 mmol) and N,N-diisopropylethylamine (81 mg, 0.63 mmol) were then added to the reaction solution in sequence, and the reaction solution was continued to react for 1 hour under stirring at 25°C. LCMS showed that the reaction was complete, and the reaction solution was filtered to obtain a filtrate, which was subjected to reverse phase column chromatography (formic acid 0.05% aqueous solution/acetonitrile) to obtain a white product A4 (21 mg, yield: 12.4%, purity: 94.5%). m/z[M+H] ⁺ =754.3. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.41(s,1H),8.97(d,J＝6.9Hz,1H),8.16(t,J＝7.9Hz,1H),8.09(d,J＝8.6Hz,1H),7.99(d,J＝2.0Hz ,1H),7.74(dd,J＝8.8,2.1Hz,1H),7.45–7.35(m,2H),6.86(s,1H),5.35(s,2H),4.55(d,J＝12.4Hz,1H ),4.28(q,J＝3.1Hz,2H),3.84(q,J＝5.4Hz,3H),3.55–3.48(m,3H),3.28–3.15(m,3H),3.02(d,J＝8.7H z,2H),2.87(t,J＝12.3Hz,1H),2.78(d,J＝10.5Hz,1H),2.62(d,J＝11.0Hz,1H),1.22(t,J＝7.4Hz,3H).

Example 5: Synthesis of Compound A5

A5-1 (50 mg, 0.088 mmol) and DMF (1 mL) were added to a reaction bottle, and HOBT (20 mg, 0.15 mmol) and EDCI (40 mg, 0.21 mmol) were added successively under stirring, and the mixture was stirred at room temperature for 1 h, which was recorded as solution 1; A4-3 (50 mg, 0.088 mmol), DIPEA (50 mg, 0.39 mmol) and acetonitrile (1 mL) were added to another reaction bottle, which was recorded as solution 2; Solution 2 was added to solution 1, and the mixture was reacted at 25°C for 2 hours. The reaction was monitored by LCMS, and A5 (4.4 mg, 7%) was obtained by reverse phase preparative separation as a white solid. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.37(s,1H),8.06(d,J＝8.6Hz,1H),7.97(s,1H),7.72(d,J＝8.8Hz,1H),6.83(s,1H),5. 31(s,2H),4.47(d,J＝12.0Hz,1H),4.25(s,2H),3.80(t,J＝5.6Hz,2H),3.68(s,4H),3.48(s, 1H),3.17(d,J＝11.7Hz,1H),2.98(s,2H),2.76(d,J＝19.2Hz,4H),2.60(d,J＝10.3Hz,1H),1. 83(d,J＝5.6Hz,2H),1.77–1.73(m,2H),1.23(s,2H),1.19(t,J＝7.5Hz,3H).LCMS(ESI)[M+1] ⁺ =758.3.

Example 6: Synthesis of Compound A6

A4-3 (50 mg, 0.088 mmol), A6-1 (20 mg, 0.13 mmol), HATU (50 mg, 0.13 mmol), DIPEA (50 mg, 0.39 mmol) and DMF (1 mL) were added to the reaction bottle and reacted at 25°C for 1 hour. The reaction was completed after LCMS monitoring, and A6 (22 mg, 35%) was separated by reverse phase preparative chromatography. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.37(s,1H),8.05(d,J＝8.6Hz,1H),7.96(d,J＝2.1Hz,1H),7.81(dd,J＝6.8,2.1Hz,1H),7.71(dd,J＝8.8,2.1Hz,1 H),7.51(dd,J＝6.9,2.0Hz,1H),6.82(dd,J＝3.2,1.6Hz,1H),6.29(t,J＝6.8Hz,1H),5.31(s,2H),4.48(d,J＝12.5Hz,1 H),4.25(q,J＝2.9Hz,2H),3.80(t,J＝5.5Hz,2H),3.47(s,3H),3.42(d,J＝8.9Hz,4H),3.29–3.18(m,2H),2.98(q,J＝7 .3Hz,2H),2.93–2.85(m,1H),2.76(d,J＝11.6Hz,1H),2.62(d,J＝10.9Hz,1H),1.18(t,J＝7.5Hz,3H).LCMS(ESI)[M+1] ⁺ =701.3.

Example 7: Synthesis of Compound A9

1) Synthesis of intermediate A9-2:

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

2) Synthesis of intermediate A9-3:

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

3) Synthesis of intermediate A9-4:

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

4) Synthesis of product A9:

Compound A1-7 (41 mg, 0.26 mmol), N-hydroxy-7-azabenzotriazole (30 mg, 0.22 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (63 mg, 0.33 mmol) were dissolved in acetonitrile (3 mL) and stirred at 25 ° C for 1 hour. Then compound A9-4 (120 mg, 0.22 mmol) and N, N-diisopropylethylamine (85 mg, 0.66 mmol) were added to the reaction solution in sequence, and the reaction solution was stirred at 25°C for 1 hour. LCMS showed that the reaction was complete, and the reaction solution was filtered to obtain a filtrate, which was purified by reverse phase column chromatography (0.05% formic acid aqueous solution/acetonitrile) to obtain a white product A9 (45 mg, yield: 27.74%, purity: 92.49%). m/z[M+H] ⁺ =680.3. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.78(s,1H),8.60(d,J＝2.8Hz,1H),7.74(d,J＝2.1Hz,1H),7.41(dd,J＝8.8,2.3Hz,1H),7.29( dd,J＝8.8,2.2Hz,1H),6.83(t,J＝2.4Hz,1H),5.19(s,2H),4.55(d,J＝12.6Hz,1H),4.31–4.23(m,2 H),3.82(t,J＝5.3Hz,2H),3.58–3.49(m,4H),3.30–3.26(m,1H),3.01(ddt,J＝17.7,14.3,6.3Hz,3 H),2.83(dd,J＝15.7,9.3Hz,2H),2.68(t,J＝9.3Hz,1H),2.47(d,J＝2.1Hz,3H),1.23–1.15(m,3H).

Example 8: Synthesis of Compound A10

Referring to the synthesis method of Experimental Example 7, Instead of A9-1, compound A10 was synthesized. m/z[M+H] ⁺ =665.4. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.60(s,1H),8.56(s,1H),7.62–7.51(m,2H),7.28(dd,J＝9.1,2.5Hz,2H),6.83–6.74(m,1H),5.13(s,2H),4.50(d,J＝12.5Hz,1H),4.22(q,J＝2.8Hz,2H),3.78(t,J＝5.4Hz,3H),3. 44(s,4H),3.24(t,J＝12.1Hz,1H),3.04–2.88(m,3H),2.81(d,J＝10.7Hz,1H),2.63(d,J＝10 .7Hz,1H),2.43(s,3H),1.69(q,J＝4.9Hz,2H),1.42(q,J＝5.1Hz,2H),1.15(t,J＝7.5Hz,3H).

Example 9: Synthesis of Compound A11

Referring to the synthesis method of Experimental Example 7, Instead of A9-1, compound A11 was synthesized. m/z[M+H] ⁺ =679.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.62(s,1H),10.26(s,1H),8.60(s,1H),7.70–7.57(m,2H),7.51–7.37(m,2H),6.83(s,1H),5.19(s,2H),4.55(d,J＝12.6Hz,1H),4.27(d,J＝3.1Hz,2H),3.82(t,J＝5.3Hz,2H),3.52(t,J＝11.3Hz,3H),3.28(s ,1H),3.00(d,J＝11.0Hz,3H),2.85(d,J＝11.0Hz,1H),2.73(ddt,J＝12.0,7.6,3.8Hz,3H),2.67–2.54(m, 4H), 2.47 (d, J＝2.0Hz, 3H), 2.38–2.22 (m, 1H), 2.02 (ddd, J＝13.8, 8.9, 4.5Hz, 1H), 1.20 (t, J＝7.5Hz, 3H).

Example 10: Synthesis of Compound A12

Referring to the synthesis method of Experimental Example 7, Instead of A9-1, compound A12 was synthesized. m/z [M+H] ⁺ = 714.3. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.31(s,1H),8.60(s,1H),7.79(d,J＝22.3Hz,2H),6.87(s,1H),5.26(s,2H),4.55(d,J＝12.5Hz,1H),4.29(s,2H),3.84(t,J＝5.6Hz,2H ),3.52(d,J＝11.5Hz,3H),3.01(q,J＝10.2,7.4Hz,4H),2.84(d,J＝11.2Hz,1H),2.66(d,J＝10.9Hz,1H),2.47(s,5H),1.22(t,J＝7.5Hz,4H).

Example 11: Synthesis of Compound A18

Referring to the synthesis method of Experimental Example 7, Instead of A9-1, compound A18 was synthesized. m/z[M+H] ⁺ =691.3. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.37(s,1H),9.49(s,1H),8.57(s,1H),8.12(d,J＝8.7Hz,1H),7.74(d,J＝8.7Hz,1H),6.86(d,J＝1.7Hz,1H),5.28(s,2H),4.52(d,J＝12.5Hz,1H),4.27(q,J＝2.8Hz,2H),3.82(q,J＝3.9Hz,2H). ＝5.5,4.7Hz,2H),3.57–3.44(m,3H),3.29–3.19(m,2H),3.01(p,J＝8.2Hz,3H),2.82(d,J＝1 0.3Hz,1H),2.64(d,J＝11.2Hz,1H),2.53(d,J＝3.3Hz,1H),2.44(s,3H),1.29–1.18(m,3H).

Example 12: Synthesis of Compound A22

Referring to the synthesis method of Experimental Example 7, Instead of A9-1, compound A22 was synthesized. m/z[M+H] ⁺ =708.4. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.61(s,1H),10.22(s,1H),8.59(s,1H),7.67–7.57(m,2H),7.43(d,J＝8.3Hz,2H),6.85–6.78(m,1H),5.18(s,2H),4.55(d,J＝12.4Hz,1H),4.26(q,J＝2.8Hz,2H),3.81(t,J＝5.4Hz,2H), 3.60–3.46(m,3H),3.29(d,J＝15.0Hz,3H),3.00(dt,J＝13.1,6.0Hz,3H),2.84(d,J＝11.7Hz,1H ),2.66(d,J＝11.0Hz,1H),2.47(s,3H),1.37–1.30(m,2H),1.19(t,J＝7.4Hz,3H),1.09(s,2H).

Example 13: Synthesis of Compound A17

Referring to the synthesis method of Experimental Example 7, Instead of A9-1, compound A17 was synthesized. m/z[M+H] ⁺ =685.3. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.52(s,1H),9.04(dd,J＝4.2,1.7Hz,1H),8.60(d,J＝2.8Hz,1H),8.46(dt,J＝8.3,2.1Hz,1H),8.01(q,J＝9.0Hz,2H),7.64(dd,J＝8.2,4.2Hz,1H),6.94-6.85(m,1H),5.38(s,2H),4.56(d,J＝12.3Hz,1H),4.29(q ,J＝2.8Hz,2H),3.83(q,J＝4.3,3.3Hz,2H),3.61–3.48(m,3H),3.30(s,2H),3.04(dd,J＝16.9,9.2Hz,3H), 2.86(d,J＝11.6Hz,1H),2.68(d,J＝10.5Hz,1H),2.55(s,1H),2.47(d,J＝2.0Hz,3H),1.25(t,J＝7.4Hz,3H).

Example 14: Synthesis of Compound A36

1) Synthesis of intermediate A36-1:

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

2) Synthesis of intermediate A36-2:

A36-1 (0.2 g, 0.3 mmol), dimethylamine hydrochloride (0.12 g, 1.5 mmol) and N,N-diisopropylethylamine (0.12 g, 0.9 mmol) were dissolved in acetonitrile (3 mL) and stirred at 80 °C for 16 hours. LCMS showed that the reaction was complete and the reaction solution was concentrated to 10% ethanol. The crude product A36-2 (0.2 g, yield: 95.1%) was obtained by condensation. ESI m/z [M+H] ⁺ = 627.3.

3) Synthesis of intermediate A36-3:

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

4) Synthesis of product A36:

Compound A1-7 (52 mg, 0.34 mmol), N-hydroxy-7-azabenzotriazole (42 mg, 0.31 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (81 mg, 0.42 mmol) were dissolved in N,N-dimethylformamide (0.5 mL) solution and reacted for 1 hour under stirring at 25°C. Subsequently, A36-3 (150 mg, 0.28 mmol) and N,N-diisopropylethylamine (110 mg, 0.84 mmol) were added to the reaction solution in sequence, and the reaction solution was stirred at 25°C for 2 hours. LCMS showed that the reaction was complete, and the reaction solution was filtered to obtain a filtrate, which was subjected to reverse phase column chromatography (formic acid 0.05% aqueous solution/acetonitrile) to obtain a white product A36 (17 mg, yield: 8.8%, purity: 97.9%). ESI m/z [M+H] ⁺ = 663.3. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.34(s,1H),8.55(s,1H),8.04(d,J＝8.6Hz,1H),7.95(d,J＝2.0Hz,1H) ,7.72(dd,J＝8.8,2.1Hz,1H),5.20(s,2H),4.50(d,J＝12.5Hz,1H),3.49(d, J＝12.4Hz,4H),3.29–3.17(m,2H),2.95(s,6H),2.93–2.88(m,1H),2.77(d, J＝11.1Hz,1H),2.59(d,J＝10.9Hz,1H),2.43(s,3H),1.15(t,J＝7.4Hz,3H).

Example 15: Synthesis of Compound A26

1) Synthesis of intermediate A26-2:

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

2) Synthesis of intermediate A26-3:

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

3) Synthesis of product A26:

Compound A1-7 (14 mg, 0.09 mmol), N-hydroxy-7-azabenzotriazole (21 mg, 0.11 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (12 mg, 0.09 mmol) were dissolved in N,N-dimethylformamide (1.5 mL) solution and reacted for 1 hour under stirring at 25°C. Compound A26-3 (50 mg, 0.07 mmol) and N,N-diisopropylethylamine (28 mg, 0.22 mmol) were then added to the reaction solution in sequence, and the reaction solution was continued to react for 1 hour under stirring at 25°C. LCMS showed that the reaction was complete, and the reaction solution was directly purified by reverse phase column chromatography to obtain a white product A26 (5.5 mg, yield: 9.2%, purity: 93.7%). ESI m/z[M+H] ⁺ =825.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.41(s,1H),8.57(s,1H),8.04(dd,J＝11.1,8.5Hz,4H),7.96(d,J＝2.0Hz,1 H),7.74–7.68(m,1H),7.08(d,J＝8.5Hz,2H),5.38(s,2H),4.53(d,J＝12.3Hz,1 H),4.15(t,J＝5.7Hz,2H),3.58(t,J＝4.7Hz,4H),3.52(d,J＝11.4Hz,4H),3.07– 2.94(m,3H),2.84(d,J＝11.2Hz,1H),2.74–2.65(m,3H),2.49–2.46(m,4H),2.44 (s,3H),1.21(s,3H).

Example 16: Synthesis of Compound A27

Reference Example 15 Synthesis of Compound A26, using Compound A27 was synthesized instead of A26-1. ESI m/z [M+H] ⁺ = 809.5. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.44(s,1H),8.61(s,1H),8.23–7.94(m,5H),7.74(dd,J＝8.8,2.1Hz,1H) ,7.11(d,J＝8.6Hz,2H),5.41(s,2H),4.56(d,J＝12.3Hz,1H),4.19(t,J＝5.7Hz ,2H),3.28(d,J＝12.4Hz,4H),3.04(q,J＝9.1,8.0Hz,4H),2.92(t,J＝5.7Hz,3 H),2.65(d,J＝5.9Hz,4H),2.48(s,3H),1.79–1.71(m,4H),1.27–1.21(m,3H).

Example 17: Synthesis of Compound A85

A85-1 (30 mg, 0.22 mmol), N-hydroxy-7-azabenzotriazole (37 mg, 0.27 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (62 mg, 0.32 mmol) were dissolved in N,N-dimethylformamide (0.5 mL) solution and reacted for 1 hour under stirring at 25°C. Compound A4-3 (100 mg, 0.18 mmol) and N,N-diisopropylethylamine (70 mg, 0.54 mmol) were then added to the reaction solution in sequence, and the reaction solution was stirred at 25°C for 1 hour. LCMS showed that the reaction was complete, and the reaction solution was filtered to obtain a filtrate, which was purified by reverse phase column chromatography (formic acid 0.05% aqueous solution/acetonitrile) to obtain a white product A85 (22 mg, yield: 17.6%, purity: 97%). ESI m/z[M+H] ⁺ =687.3. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.40(s,1H),8.08(d,J＝8.6Hz,1H),7.99(d,J＝2.1Hz,1H),7.87(s,1H),7.81–7.6 5(m,2H),6.93–6.81(m,1H),6.22(s,1H),5.34(s,2H),4.51(d,J＝12.6Hz,1H),4.28(p ,J＝3.2Hz,2H),3.83(t,J＝5.5Hz,2H),3.45(s,4H),3.26(s,1H),3.01(q,J＝7.4Hz,2H) ,2.95–2.73(m,3H),2.66(d,J＝10.5Hz,1H),2.15–1.73(m,1H),1.22(t,J＝7.4Hz,3H).

Example 18: Synthesis of Compound A79

1) Synthesis of intermediate A79-2:

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

2) Synthesis of intermediate A79-3:

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

3) Synthesis of product A79:

Compound A1-7 (28 mg, 0.18 mmol), N-hydroxy-7-azabenzotriazole (21 mg, 0.15 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (40 mg, 0.21 mmol) were dissolved in N,N-dimethylformamide (3 mL) solution and reacted for 1 hour under stirring at 25°C. Compound A79-3 (86 mg, 0.14 mmol) and N,N-diisopropylethylamine (54 mg, 0.42 mmol) were then added to the reaction solution in sequence, and the reaction solution was stirred at 25°C for 2 hours. LCMS showed that the reaction was complete, and the reaction solution was filtered to obtain a filtrate, which was subjected to reverse phase column chromatography (formic acid 0.05% aqueous solution/acetonitrile) to obtain a white product A79 (32 mg, yield: 30.4%, purity: 99.8%). ESI m/z[MH] ^- =748.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.43(s,1H),8.58(s,1H),8.53–8.47(m,2H),8.07(d,J＝8.6Hz,1H),8.0 0–7.94(m,2H),7.74–7.66(m,2H),5.41(s,2H),4.54(d,J＝12.0Hz,1H),4.19 (s,3H),3.52(t,J＝10.6Hz,3H),3.27(s,1H),3.02(d,J＝9.2Hz,3H),2.85(d ,J＝11.2Hz,1H),2.67(d,J＝10.6Hz,1H),2.45(s,3H),1.21(t,J＝5.6Hz,3H).

Example 19: Synthesis of Compound A76

Reference Example 18 Synthesis of Compound A79, using Instead of A79-1, compound A76 was synthesized. ESI m/z[MH] ^- =748.5. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.46(s,1H),8.61(s,1H),8.56(d,J＝1.4Hz,1H),8.24-8.15(m,2H),8.10(d,J＝8.6Hz,1H),8.00(d,J＝2.0Hz,1H),7.79(d,J＝8.8Hz,1H),7.74(dd,J＝8.7,2.2Hz,1H),5.44(s ,2H),4.57(d,J＝12.4Hz,1H),4.11(s,3H),3.57(q,J＝10.2,8.6Hz,3H),3.04(t,J＝11.0 Hz,3H),2.88(d,J＝11.3Hz,1H),2.70(d,J＝10.5Hz,1H),2.48(s,3H),1.28–1.24(m,3H).

Example 20: Synthesis of Compound A78

Reference Example 18 Synthesis of Compound A79, using Compound A78 was synthesized by replacing A79-1. ESI m/z[MH] ^- =735.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.44(s,1H),9.41–9.34(m,2H),8.61(s,1H),8.11(d,J＝8.6Hz,1H),8.03– 7.99(m,2H),7.94(d,J＝9.6Hz,1H),7.74(dd,J＝8.8,2.1Hz,1H),5.43(s,2H),4 .57(d,J＝12.1Hz,1H),3.55(d,J＝11.6Hz,3H),3.30(s,1H),3.10–3.00(m,3H), 2.92–2.85(m,1H),2.71(d,J＝10.7Hz,1H),2.47(s,3H),1.24(d,J＝7.7Hz,3H).

Example 21: Synthesis of Compound A19

1) Synthesis of intermediate A19-2:

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

2) Synthesis of intermediate A19-3:

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

3) Synthesis of intermediate A19-4:

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

4) Synthesis of intermediate A19-5:

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

5) Synthesis of intermediate A19-6:

Compound A19-5 (0.3 g, 1.45 mmol) and potassium carbonate (0.6 g, 4.35 mmol) were dissolved in dichloromethane (2 mL) solution, bromoacetyl bromide (0.44 g, 2.17 mmol) was added dropwise under stirring at 0°C, and stirred at 25°C for 1 hour. LCMS showed that the reaction was complete, the reaction solution was poured into water, extracted with dichloromethane (50 mL*2), and concentrated to obtain yellow solid A19-6 (0.3 g, mixture, yield: 63.2%). Directly proceed to the next step.

6) Synthesis of intermediate A19-7:

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

7) Synthesis of intermediate A19-8:

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

8) Synthesis of product A19:

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

A19: ESI m/z[M+H] ⁺ =714.3. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.36(s,1H),8.54(s,1H),7.44(d,J＝2.2Hz,2H),6.88(d,J＝2.8Hz,1H),5.24(s,2H),4.54( d,J＝12.5Hz,1H),4.29(q,J＝2.8Hz,2H),3.84(t, J＝5.4Hz,2H),3.57–3.49(m,4H),3.01(q,J＝8.6,8.1Hz,4H),2.83(d,J＝11.4Hz,2H),2.65(d,J＝11.0 Hz,1H),2.45(s,3H),1.22(t,J=7.5Hz,3H).

A19': ESI m/z[M+H] ⁺ =714.3. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.85(s,1H),8.59(s,1H),7.60(d,J＝9.2Hz,1H),7.34(d,J＝9.2Hz,1H),6.9 3–6.81(m,1H),5.25(s,2H),4.55(d,J＝12.6Hz,1H),4.27(q,J＝2.9Hz,2H),3.83 (t,J＝5.4Hz,2H),3.53(d,J＝12.1Hz,3H),3.28(s,3H),3.00(t,J＝7.7Hz,3H),2. 84(d,J＝11.4Hz,1H),2.67(t,J＝9.5Hz,1H),2.47(s,3H),1.20(t,J＝7.5Hz,3H).

Example 22: Synthesis of Compound A77

Reference Example 18 Synthesis of Compound A79, using Instead of A79-1, compound A77 was synthesized. ESI m/z[MH] ^- =734.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ13.33(s,1H),10.47(s,1H),8.61(s,1H),8.30(s,1H),8.18(s,1H),8.09(d,J＝8.6Hz,1H),8.00(d,J＝2.1Hz,1H),7.92(s,2H),7.74(dd,J＝8.7,2.1Hz,1H),5.45(s ,2H),4.57(d,J＝12.5Hz,1H),3.56(t,J＝10.8Hz,3H),3.29(d,J＝12.9Hz,2H),3.09 –3.03(m,2H),2.89(d,J＝11.2Hz,1H),2.78–2.66(m,1H),2.48(s,3H),1.25(s,3H).

Example 23: Synthesis of Compound A75

Reference Example 14 Synthesis of Compound A36 using Instead of dimethylamine, compound A75 was synthesized. ESI m/z [M+H] ⁺ = 720.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.36(s,1H),8.59(s,1H),8.08(d,J＝8.6Hz,1H),7.98(d,J＝2.1Hz,1H),7. 74(dd,J＝8.8,2.1Hz,1H),5.23(s,2H),4.53(d,J＝12.4Hz,1H),3.53(s,4H),3 .27(d,J＝11.7Hz,3H),2.99(s,6H),2.80(d,J＝11.5Hz,1H),2.62(d,J＝10.7Hz ,1H),2.49(d,J＝4.6Hz,1H),2.46(s,3H),2.18(s,6H),1.18(t,J＝7.4Hz,3H).

Example 24: Synthesis of Compound A20

1) Synthesis of intermediate A20-2:

A20-1 (9.0 g, 44.53 mmol) and anhydrous tetrahydrofuran (225 mL) were added to a 1000 mL three-necked flask in sequence, stirred at -70 to -75 °C under nitrogen protection, and a n-butyl lithium hexane solution (2.5 mol/L, 48 mL, 120 mmol) was slowly added, stirred at -70 to -75 °C for 20 minutes, slowly heated to -5 to 0 °C, stirred at -5 to 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, stirred at -70 to -75 °C for 40 minutes, then slowly returned to room temperature and continued to stir for 40 minutes. The reaction was quenched with a 10% mass fraction citric acid aqueous solution, extracted twice with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and column chromatography was performed to obtain A20-2 (3.5 g, 33%).

2) Synthesis of intermediate A20-3:

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

3) Synthesis of intermediate A20-4:

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

4) Synthesis of intermediate A20-5:

Compound A20-4 (0.3 g, 1.45 mmol) and potassium carbonate (0.6 g, 4.35 mmol) were dissolved in dichloromethane (2 mL) solution, bromoacetyl bromide (0.44 g, 2.17 mmol) was added dropwise under stirring at 0°C, and the mixture was stirred at 25°C for 1 hour. LCMS showed that the reaction was complete, and the reaction solution was poured into water, extracted with dichloromethane (50 mL*2), and concentrated to obtain yellow solid A20-5 (0.3 g, 63%).

5) Synthesis of intermediate A20-6:

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

6) Synthesis of intermediate A20-7:

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

7) Synthesis of intermediate A20-8:

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

8) Synthesis of product A20:

A1-7 (1 g, 6.49 mmol), HOBT (0.88 g, 6.43 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (1.54 g, 8.04 mmol) were dissolved in N, N-dimethylformamide (2 mL) solution and reacted for 1 hour under stirring at 25°C. Compound A20-8 (3 g, 5.36 mmol) and N, N-diisopropylethylamine (2.08 g, 16.08 mmol) were then added to the reaction solution in sequence, and the reaction solution was continued to react for 1 hour under stirring at 25°C. LCMS showed that the reaction was complete, and the reaction solution was added to water, extracted with dichloromethane, washed with water, washed with saturated brine, dried with sodium sulfate, filtered, concentrated, and purified by silica gel column (dichloromethane/ethyl acetate) to obtain a white product A20 (1.45 g, yield: 38.88%, purity: 97.82%). m/z[M+H] ⁺ =708.4. ¹ HNMR (400MHz, DMSO-d ₆ )δ10.39(s,1H),8.60(s,1H),8.15(dd,J＝6.8,3.0Hz,2H),7.56(dd,J＝5.1,2.0Hz,3H),7.44(s,2H),5.32(s,2H),4.56(d,J＝12.6Hz,1H),3. 55(d,J＝12.0Hz,3H),3.27(s,1H),3.04(dd,J＝13.5,9.0Hz,3H),2.90–2.81(m,1H),2.69(d,J＝10.7Hz,1H),2.47(s,3H),1.32–1.24(m,3H).

Example 25: Synthesis of Compound A3

Referring to the synthesis of compound A26 in Example 15, 2-methyl-2H-indazole-6-boronic acid was used instead of A26-1 to synthesize compound A3. m/z[M+H] ⁺ =750.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.48(s,1H),8.61(s,1H),8.44(s,1H),8.37(d,J＝1.2Hz,1H),8.08(d,J＝8.6Hz,1H),8.00(d,J＝2.1Hz,1H),7.89-7.80(m,2H),7.74(dd,J＝8.8,2.1Hz,1H),5.45(s,2H),4.57 (d,J＝12.4Hz,1H),4.23(s,3H),3.57(td,J＝12.4,11.8,6.6Hz,4H),3.05(dd,J＝15.8,9. 5Hz,3H),2.88(d,J＝11.3Hz,1H),2.76–2.67(m,1H),2.48(s,3H),1.26(d,J＝7.3Hz,3H).

Example 26: Synthesis of Compound A7

Referring to the synthesis of compound A26 in Example 15, 1-methyl-1H-indazole-6-boronic acid was used instead of A26-1 to synthesize compound A7. m/z[MH] ^- =748.5. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.50(s,1H),8.62(s,1H),8.35(s,1H),8.15(s,1H),8.11(d,J＝8.6Hz,1H),8.01(d,J＝2.0Hz,1H),7.98-7.90(m,2H),7.76(dd,J＝8.8,2.1Hz,1H),5.48(s,2H),4.58(d,J＝1 2.5Hz,1H),4.15(s,3H),3.58(q,J＝11.7,10.8Hz,3H),3.28(s,1H),3.07(d,J＝9.0Hz,3 H), 2.90 (d, J = 11.3Hz, 1H), 2.72 (d, J = 11.0Hz, 1H), 2.49 (s, 3H), 1.25 (d, J = 7.6Hz, 3H).

Example 27: Synthesis of Compound A8

Referring to the synthesis of compound A26 in Example 15, (2,2-difluorobenzo[d][1,3]dioxol-5-yl)boronic acid was used instead of A26-1 to synthesize compound A8. m/z[MH] ^- =774.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.45(s,1H),8.60(s,1H),8.08(d,J＝8.6Hz,1H),8.04(d,J＝1.5Hz,1H),8.03-7.99(m,2H),7.74(dd,J＝8.8,2.1Hz,1H),7.60(d,J＝8.4Hz,1H),5.42(s,2H),4.56(d,J＝1 2.3Hz,1H),3.56(dd,J＝14.1,10.7Hz,3H),3.32–3.25(m,2H),3.03(dd,J＝14.8,8.4H z,3H),2.87(d,J＝11.1Hz,1H),2.72–2.66(m,1H),2.47(s,3H),1.24(t,J＝7.4Hz,3H).

Example 28: Synthesis of Compound A21

Referring to the synthesis of compound A1 in Example 1, compound A21 was synthesized by using A20-5 instead of A1-4. m/z[M+H] ⁺ =806.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.37(s,1H),8.55(s,1H),7.94(d,J＝8.6Hz,2H),7.41(s,2H),7.04(d,J ＝8.6Hz,2H),5.27(s,2H),4.53(d,J＝12.3Hz,1H),3.53(d,J＝3.1Hz,2H),3. 49(d,J＝11.2Hz,4H),3.27–3.25(m,4H),3.01(d,J＝8.8Hz,4H),2.82(d,J＝1 1.1Hz,2H),2.71–2.61(m,2H),2.44(s,3H),2.23(s,3H),1.23–1.19(m,3H).

Example 29: Synthesis of Compound A30

1) Synthesis of intermediate A30-2:

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

2) Synthesis of intermediate A30-3:

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

3) Synthesis of product A30:

Referring to the synthesis of compound A9 in Example 7, compound A30 was synthesized by using A30-3 instead of A9-1. m/z[M+H] ⁺ =666.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.18(s,1H),8.58(s,1H),7.78(s,1H),6.92-6.80(m,1H),5.19 (s,2H),4.64(p,J＝6.6Hz,1H),4.54(d,J＝12.5Hz,1H),4.28(p,J＝2.9Hz,2H),3.83(t,J＝5.3Hz,2H),3.60(s,2H),3.27(d,J＝2.7Hz,4H) ,2.99(d,J＝12.4Hz,3H),2.84(d,J＝10.4Hz,1H),2.66(d,J＝10.9Hz,1H),2.46(s,3H),1.40(s,3H),1.38(s,3H),1.20(t,J＝7.5Hz,3H).

Example 30: Synthesis of Compound A32

Referring to the synthesis of compound A31 in Example 3, compound A32 was synthesized by replacing A31-1 with A32-1. m/z[MH] ^- =702.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.39(s,1H),8.99(s,1H),8.09(d,J＝8.3Hz,2H),7.99(s,1H),6.86(s,1H),5.35(s,2H),4.28(s,3H),3.82(t,J＝5.1Hz,3H),3.74(s,3H),3.48(d,J＝11.1Hz,4H),3.08-2.98(m,4H),2.79(s,2H),2.17(s,3H),1.23(s,3H).

Example 31: Synthesis of Compound A47

Referring to the synthesis of compound A36 in Example 14, 2-oxa-6-azaspiro[3.3]heptane was used instead of dimethylamine to synthesize compound A47. m/z[M+H] ⁺ =717.3. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.38(s,1H),8.54(s,1H),8.09(d,J＝8.6Hz,1H),7.99(d,J＝2.1Hz,1H),7.7 5(dd,J＝8.8,2.2Hz,1H),5.24(s,2H),4.72(s,3H),4.53(d,J＝12.4Hz,1H),4.16 (s,3H),3.52(s,6H),2.97(q,J＝10.4,7.8Hz,3H),2.79(d,J＝11.5Hz,1H),2.72 –2.57(m,1H),2.45(s,2H),2.02(dt,J＝13.6,7.0Hz,1H),1.17(t,J＝7.4Hz,3H).

Example 32: Synthesis of Compound A70

1) Synthesis of intermediate A70-1:

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

2) Synthesis of product A70:

Referring to the synthesis of compound A1 in Example 1, phenylboronic acid was used instead of A1-2, and A70-1 was used instead of A1-4 to synthesize compound A70. m/z[M+H] ⁺ =687.2. ¹ H NMR (400MHz, DMSO-d ₆ )δ8.46(s,1H),8.08(d,J＝5.9Hz,3H),7.70(d,J＝7.7Hz,1H),7.57-7.46(m,4H),7.04(d,J＝7.7Hz,1H),5.42(s,2H),5.08–4.97(m,1H), 3.54-3.48(m,2H),3.04-2.94(m,6H),2.86-2.80(m,1H),2.69-2.62(m,1H),2.40(s,3H),1.26(d,J＝6.7Hz,6H),1.17(t,J＝7.2Hz,3H).

Example 33: Synthesis of Compound A71

1) Synthesis of intermediate A71-2:

A71-1 (950 mg, 6.78 mmol) and potassium carbonate (1.88 g, 13.4 mmol) were dissolved in acetonitrile (10 mL), 2-iodopropane (5.77 g, 33.9 mmol) was added under ice bath, and stirred at this temperature for 1 hour. After LCMS showed that the reaction was completed, it was slowly poured into water (10 mL) and extracted with ethyl acetate (30 mL×3). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by evaporation under reduced pressure. The residue was purified by silica gel column chromatography to obtain a yellow oil A71-2 (910 mg, yield 73.0%). m/z[M+H] ⁺ =183.1. ¹ HNMR (400MHz, DMSO-d ₆ )δ(ppm)8.09(d,J＝7.6Hz,1H),7.11(d,J＝2.7Hz,1H),6.88(dd,J＝7.6,2.7Hz,1H),5.03(p,J＝6.8Hz,1H),1.33(d,J＝6.8Hz,6H).

2) Synthesis of intermediate A71-3:

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

3) Synthesis of intermediate A71-4:

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

4) Synthesis of product A71:

Referring to the synthesis of compound A1 in Example 1, phenylboronic acid was used instead of A1-2, and A71-4 was used instead of A1-4 to synthesize compound A71. m/z[M+H] ⁺ =687.2. ¹ H NMR (400 MHz, DMSO-d ₆ )δ11.80(s,1H),9.52(s,1H),8.59(s,1H),8.22(s,2H),7.47(s,3H),6.73(d,J＝7.7Hz,1H),6.60(s,1H),5.73-5.61(m,1H),5.25-5.05(m ,3H),3.83(t,J=11.6Hz,2H),3.55-3.42(m,1H),3.19-2.74(m,4H),2.57(s,3H),2.34-2.17(m,1H),2.04-1.96(m,1H),1.33-1.28(m,9H).

Example 34: Synthesis of Compound A81

Referring to the synthesis of compound A1 in Example 1, 1-methyl-1H-indazole-5-boronic acid was used instead of A1-2, and A20-5 was used instead of A1-4 to synthesize compound A81. m/z[MH] ^- ＝760.4. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.42(s,1H),8.67–8.54(m,2H),8.24–8.13(m,2H),7.81(d,J＝8.9Hz, 1H),7.45(s,2H),5.34(s,2H),4.57(d,J＝12.4Hz,1H),4.12(s,3H),3.56 (d,J＝11.2Hz,3H),3.30–3.24(m,1H),3.05(d,J＝10.1Hz,3H),2.87(d,J＝ 11.1Hz,1H),2.69(d,J＝10.3Hz,1H),2.47(s,3H),1.27(d,J＝7.1Hz,3H).

Example 35: Synthesis of Compound A82

Referring to the synthesis of compound A1 in Example 1, 2-methyl-2H-indazole-5-boronic acid was used instead of A1-2, and A20-5 was used instead of A1-4 to obtain compound A82. m/z[MH] ^- =760.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.41(s,1H),10.27(s,1H),8.61(s,1H),8.58-8.52(m,2H),8.01(dd,J＝9.1,1.6Hz,1H),7.74(d,J＝9.1Hz,1H),7.45(s,2H),5.33(s,2H),4.57(d,J＝12.5Hz,1H) ,4.23(s,3H),3.64–3.47(m,3H),3.28(d,J＝12.7Hz,1H),3.05(d,J＝9.3Hz,3H),2 .87(d,J＝11.0Hz,1H),2.69(d,J＝10.4Hz,1H),2.48(s,3H),1.25(d,J＝7.9Hz,3H).

Example 36: Synthesis of Compound A91

Referring to the synthesis of compound A26 in Example 15, quinoline-6-pinacol boronate was used instead of A26-1 to synthesize compound A91. m/z[MH] ^- =745.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.48(s,1H),9.00(dd,J＝4.2,1.7Hz,1H),8.79(d,J＝1.9Hz,1H),8.62(s,1H),8.58(dd,J＝8.4,1.7Hz,1H),8.49(dd,J＝8.8,1.9Hz,1H),8.18(d,J＝8.8Hz,1H),8.12(d,J＝8.6Hz,1H),8.01(d,J＝2.1Hz,1H),7.74(dd,J＝8.9,2. 1Hz,1H),7.64(dd,J＝8.3,4.2Hz,1H),5.48(s,2H),4.58(d,J＝12.5Hz,1H),3.58(td,J＝12.6,11.7,7.0Hz,3H),3. 28(s,2H),3.06(p,J＝9.3,8.4Hz,3H),2.90(d,J＝11.3Hz,1H),2.80–2.67(m,1H),2.48(s,3H),1.28–1.24(m,3H).

Example 37: Synthesis of Compound A92

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

Example 38: Synthesis of Compound A136

Referring to the synthesis of compound A26 in Example 15, pyrazolo[1,5-a]pyridine-5-pinacol boronate was used instead of A26-1 to synthesize compound A136. m/z[MH] ^- =734.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.39(s,2H),8.77(d,J＝7.3Hz,1H),8.56(s,1H),8.40(s,1H),8.15-8.01(m,2H),7.94(d,J＝2.1Hz,1H),7.69(dd,J＝9.1,2.0Hz,1H),7.49(dd,J＝7.3,1.8Hz,1H),6.80(d,J＝2 .2Hz,1H),5.39(s,2H),4.52(d,J＝12.6Hz,1H),3.52(d,J＝11.4Hz,3H),3.01(d,J＝8.8Hz ,3H),2.84(d,J＝11.3Hz,1H),2.66(d,J＝10.8Hz,1H),2.43(s,3H),1.20(d,J＝7.8Hz,3H).

Example 39: Synthesis of Compound A139

Referring to the synthesis of compound A26 in Example 15, benzo[b]thiophene-2-boronic acid was used instead of A26-1 to synthesize compound A139. m/z[MH] ^- =750.3. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.45(s,1H),8.61(s,1H),8.16(s,1H),8.09(dd,J＝11.5,7.5Hz,2H),8.00(q,J＝4.3,3.5Hz,2H),7.74(dd,J＝8.8,2.1Hz,1H),7.52-7.43(m,2H),5.42(s,2H),4.57(d,J＝ 12.4Hz,1H),3.55(dd,J＝11.8,3.3Hz,3H),3.29(d,J＝17.3Hz,2H),3.05(d,J＝9.9Hz,3 H), 2.88 (d, J = 11.2Hz, 1H), 2.70 (d, J = 10.2Hz, 1H), 2.47 (s, 3H), 1.24 (t, J = 7.3Hz, 3H).

Example 40: Synthesis of Compound A141

1) Synthesis of intermediate A141-4:

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

2) Synthesis of intermediate A141-5:

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

2) Synthesis of product A141:

Referring to the synthesis of compound A26 in Example 15, compound A141 was synthesized by replacing A36-1 with A141-5 and replacing A26-1 with phenylboronic acid. m/z[M+H] ⁺ =692.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.56(s,1H),8.57(s,1H),8.13(dd,J＝6.6,3.0Hz,2H),7.60–7.48(m,4H),7.29(d,J＝8.8Hz,1H),5.32(s,2H),4.56(d,J＝12.6Hz,1H),3.5 4(d,J＝10.5Hz,3H),3.03(q,J＝8.3Hz,3H),2.86(d,J＝11.3Hz,1H),2.71–2.66(m,1H),2.46(s,3H),2.10-1.96(m,1H),1.23(t,J＝7.4Hz,3H).

Example 41: Synthesis of Compound A142

1) Synthesis of intermediate A142-2:

Compound A142-1 (0.6 g, 2.38 mmol) and methylboric acid (1.42 g, 23.8 mmol) were dissolved in a mixed solution of 1,4-dioxane (5 mL) and water (0.5 mL), potassium carbonate (0.99 g, 7.14 mmol) was added under stirring, 1,1-bis(diphenylphosphino)ferrocenepalladium dichloride (II) (0.17 g, 0.24 mmol) was added after replacing the gas three times, and the reaction was 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 a white solid A142-2 (0.4 g, yield: 89.8%). m/z[M+H] ⁺ =188.1.

2) Synthesis of intermediate A142-3:

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

3) Synthesis of product A142:

Referring to the synthesis of compound A1 in Example 1, phenylboronic acid was used instead of A1-2, and A142-3 was used instead of A1-4 to synthesize compound A142. m/z[M+H] ⁺ =688.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.14(s,1H),8.61(s,1H),8.19–8.10(m,2H),7.57(dd,J＝5.2,2.0Hz,3H),7. 24(d,J＝8.7Hz,1H),7.16(d,J＝8.7Hz,1H),5.28(s,2H),4.57(d,J＝12.5Hz,1H), 3.60–3.53(m,3H),3.30(s,1H),3.06(td,J＝13.5,12.0,8.4Hz,3H),2.87(d,J＝1 0.7Hz,1H),2.73–2.66(m,1H),2.48(s,3H),2.22(s,3H),1.26(d,J＝3.4Hz,3H).

Example 42: Synthesis of Compound A143

1) Synthesis of intermediate A143-1:

Refer to the synthesis of compound A9-2 in Example 7, and replace A9-1 with A142-1 to obtain A143-1.

3) Synthesis of product A143:

Referring to the synthesis of compound A1 in Example 1, phenylboronic acid was used instead of A1-2, and A143-1 was used instead of A1-4 to synthesize compound A143. m/z [M+H] ⁺ = 752.1, 754.1. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.35(s,1H),8.59(s,1H),8.16(dd,J＝6.9,2.9Hz,2H),7.56(dd,J＝5.2,1 .9Hz,3H),7.46(d,J＝8.7Hz,1H),7.37(d,J＝8.7Hz,1H),5.31(s,2H),4.56(d ,J＝12.6Hz,1H),3.54(d,J＝13.8Hz,3H),3.03(dd,J＝15.4,9.4Hz,3H),2.87( d,J＝11.3Hz,1H),2.69(d,J＝10.6Hz,1H),2.47(s,3H),1.27(d,J＝7.0Hz,3H).

Example 43: Synthesis of Compound A144

1) Synthesis of intermediate A144-1:

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

2) Synthesis of product 144:

Referring to the synthesis of compound A26 in Example 15, compound A144 was synthesized by replacing A36-1 with A144-1 and replacing A26-1 with phenylboronic acid. m/z [M+H] ⁺ = 674.4. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.83(s,1H),8.59(s,1H),8.11(dd,J＝6.6,3.0Hz,2H),7.75(d,J＝2.0Hz ,1H),7.61–7.52(m,3H),7.41(d,J＝8.8Hz,1H),7.30(dd,J＝8.8,2.1Hz,1H) ,5.26(s,2H),4.56(d,J＝12.6Hz,1H),3.66–3.55(m,4H),3.06–2.97(m,3H) ,2.87(d,J＝11.3Hz,1H),2.70(d,J＝10.4Hz,1H),2.47(s,3H),1.22(s,3H).

Example 44: Synthesis of Compound A145

1) Synthesis of intermediate A145-1:

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

2) Synthesis of product 145:

Referring to the synthesis of compound A26 in Example 15, A145-1 was used instead of A36-1, and thiophene-2-pinacol boronate was used instead of A26-1 to synthesize compound A145. m/z [M+H] ⁺ = 714.3. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.39(s,1H),8.59(s,1H),7.84–7.76(m,2H),7.44(s,2H),7.24(dd,J＝5.0,3.6Hz,1H),5.28(s,2H),4.56(d,J＝12.5Hz,1H),3.58–3.50(m ,3H),3.29(d,J＝2.9Hz,1H),3.04(q,J＝11.8,9.1Hz,3H),2.86(d,J＝11.2Hz,1H),2.68(d,J＝11.2Hz,1H),2.47(s,3H),1.24(t,J＝7.5Hz,3H).

Example 45: Synthesis of Compound A146

Referring to the synthesis of compound A26 in Example 15, A145-1 was used instead of A36-1, and furan-2-pinacol boronate was used instead of A26-1 to synthesize compound A146. m/z [MH] ^- = 696.4. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.38(s,1H),8.60(s,1H),7.94(s,1H),7.44(s,2H),7.17(d,J＝3.4Hz,1H),6.76–6.69(m,1H),5.29(s,2H),4.56(d,J＝12.5Hz,1H),3.53 (d,J＝10.6Hz,3H),3.32–3.27(m,1H),3.03(q,J＝8.1Hz,3H),2.87–2.83(m,1H),2.68(d,J＝11.0Hz,1H),2.47(s,3H),1.24(d,J＝6.7Hz,3H).

Example 46: Synthesis of Compound A147

Referring to the synthesis of compound A26 in Example 15, A145-1 was used instead of A36-1, and 4-dimethylaminophenylboronic acid was used instead of A26-1 to obtain compound A147. ESI (m/z) = 751.4 [M+H] ⁺ . ¹ H NMR (400 MHz, chloroform-d) δ 11.80 (s, 1H), 9.17 (s, 1H), 8.59 (s, 1H), 8.17-8.06 (m, 2H), 8.00 (d, J = 8.9 Hz, 1H), 6.96 (d, J = 8.9 Hz, 1H), 6.73 (d, J = 8.9 Hz, 2H), 5.66 (d, J = 13.0 Hz, 1H), 5.12(s,2H),4.79(d,J＝12.8Hz,1H),3.83(dd,J＝13.1,10.2Hz,2H),3.50(s,1H),3.21 (q,J＝7.6Hz,2H),3.04(s,7H),2.90–2.71(m,2H),2.57(s,3H),1.34(t,J＝7.5Hz,3H).

Example 47: Synthesis of Compound A148

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

Example 48: Synthesis of Compound A149

Referring to the synthesis of compound A26 in Example 15, compound A149 was synthesized by replacing A36-1 with A145-1 and A26-1 with 4-methoxyphenylboronic acid. m/z [MH] ^- = 736.5. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.40(s,1H),8.60(s,1H),8.10–8.06(m,2H),7.44(s,2H),7.15–7.08(m,2H),5.30(s,2H),4.56(d,J＝12.4Hz,1H),3.86(s, 3H),3.53(s,3H),3.27(s,1H),3.04(d,J＝9.2Hz,3H),2.86(d,J＝10.6Hz,1H),2.68(d,J＝11.5Hz,1H),2.47(s,3H),1.24(s,3H).

Example 50: Synthesis of Compound A151

Referring to the synthesis of compound A26 in Example 15, 1-methyl-1H-indole-5-boronic acid pinacol ester was used instead of A26-1 to synthesize compound A151. m/z[M+H] ⁺ =749.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.44(s,1H),8.58(s,1H),8.38–8.33(m,1H),8.07(d,J＝8.5Hz,1H),8.01–7.90(m,2H),7.71(d,J＝8.6Hz,1H),7.55(d,J＝8.7Hz,1H),7.40(d,J＝3.1Hz,1H),6.54(d,J＝3.1Hz,1H),5.41( s,2H),4.54(d,J＝12.4Hz,1H),3.83(s,3H),3.55(q,J＝11.3Hz,3H),3.25(s,1H),3.02(d,J＝1 1.0Hz, 3H), 2.85 (d, J = 11.3Hz, 1H), 2.67 (d, J = 9.4Hz, 1H), 2.45 (s, 3H), 1.22 (t, J = 7.3Hz, 3H).

Example 51: Synthesis of Compound A152

Referring to the synthesis of compound A26 in Example 15, 1-methyl-1H-benzo[d]imidazole-5-boronic acid pinacol ester was used instead of A26-1 to synthesize compound A152. m/z[M+H] ⁺ =750.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.95(s,1H),9.06(s,1H),8.86(s,1H),8.76(s,1H),8.55(t,J＝8.9Hz,2 H),8.45(s,1H),8.18(d,J＝8.6Hz,2H),5.90(s,2H),5.02(d,J＝12.2Hz,1H), 4.36(s,3H),4.01(s,3H),3.75(s,1H),3.50(p,J＝10.6,9.1Hz,3H),3.33(d ,J＝11.3Hz,1H),3.15(d,J＝10.7Hz,1H),2.93(s,3H),1.70(t,J＝7.4Hz,3H).

Example 52: Synthesis of Compound A153

Referring to the synthesis of compound A26 in Example 15, compound A153 was synthesized by using benzo[d]thiazole-5-boronic acid pinacol ester instead of A26-1. m/z[MH] ^- =751.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.48(s,1H),9.50(s,1H),8.76(d,J＝1.5Hz,1H),8.58(s,1H),8.34(d,J＝8.4Hz,1H),8.23(dd,J＝8.4,1.6Hz,1H),8.06(d,J＝8.6Hz,1H),7.98(d,J＝2.0Hz,1H),7.72(dd,J＝ 8.8,2.1Hz,1H),5.44(s,2H),4.55(d,J＝12.6Hz,1H),3.53(t,J＝11.4Hz,4H),3.11–3.0 0(m,3H),2.86(d,J＝11.3Hz,1H),2.74–2.65(m,1H),2.45(s,3H),1.24(d,J＝6.9Hz,3H).

Example 53: Synthesis of Compound A154

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

Example 54: Synthesis of Compound A155

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

Example 55: Synthesis of Compound A156

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

Example 56: Synthesis of Compound A157

Reference Example 15 Synthesis of Compound A26, using A145-1 instead of A36-1, and using 4-(4-(oxetan-3-yl)piperazin-1-yl)phenylboronic acid pinacol ester instead of A26-1 to synthesize Compound A157. ESI (m/z) = 848.5 [M+H] ⁺ . ¹ H NMR (400 MHz, chloroform-d) δ 9.07 (s, 1H), 8.58 (s, 1H), 8.14 (d, J = 8.8 Hz, 2H), 7.99 (d, J = 8.9 Hz, 1H), 6.95 (t, J = 8.6 Hz, 3H), 5.66 (d, J = 12.9 Hz, 1H), 5.13 (s, 2H), 4.79 (d, J = 12.8 Hz, 1H), 4.69 (dt, J = 16.7, 6.3H z,4H),3.82(t,J＝11.4Hz,2H),3.61–3.42(m,2H),3.36(t,J＝4.9Hz,4H),3.21(q,J＝7.5Hz,2H),3.0 7(s,1H),2.80(dd,J＝32.8,11.5Hz,2H),2.56(s,3H),2.50(t,J＝5.0Hz,4H),1.34(t,J＝7.5Hz,3H).

Example 57: Synthesis of Compound A158

Referring to the synthesis of compound A26 in Example 15, A145-1 was used instead of A36-1, and 6-(4-methylpiperazin-1-yl)pyridine-3-boronic acid pinacol ester was used instead of A26-1 to obtain compound A158. m/z[M+H] ⁺ =807.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.36(s,1H),8.79(d,J＝2.4Hz,1H),8.57(s,1H),8.19-8.08(m,1H),7.40(s,2H),6.96(d,J＝9.1Hz,1H),5.26(s,2H),4.53(d,J＝12.3Hz,1H),3.60(t,J＝5.1Hz,4H),3.5 2(s,2H),3.26–3.23(m,2H),2.99(t,J＝8.6Hz,3H),2.82(d,J＝11.1Hz,1H),2.64(d,J ＝11.0Hz,1H),2.44(s,3H),2.41(t,J＝5.0Hz,4H),2.23(s,3H),1.21(t,J＝7.5Hz,3H).

Example 58: Synthesis of Compound A160

1) Synthesis of intermediate A160-2:

Add diisopropylethylamine (170 mg, 12.33 mmol) and A20-5 (1.49 g, 4.52 mmol) to a solution of A160-1 (1 g, 4.11 mol) in acetonitrile (10 mL), and stir overnight at room temperature. The reaction solution was dried by rotary evaporation and separated by column chromatography to obtain A160-2 (1.6 g, 80%).

2) Synthesis of intermediate A160-3:

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

3) Synthesis of intermediate A160-4:

Add liquid bromine (0.74 g, 4.65 mmol) to a solution of A160-3 (0.5 g, 0.93 mmol) in tetrahydrofuran (5 mL). Stir for 4 hours. LCMS showed that the reaction was complete. The reaction solution was filtered to obtain a yellow solid product A160-4 (0.5 g, yield: 87%). m/z[M+H] ⁺ =617.2.

4) Synthesis of intermediate A160-6:

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

5) Synthesis of product A160:

Referring to the synthesis of compound A1 in Example 1, compound A160 was synthesized by replacing A1-5 with A160-6. m/z[M+H] ⁺ =785.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.43(s,1H),9.00(dd,J＝4.2,1.7Hz,1H),8.84-8.77(m,1H),8.59(d,J＝7.4Hz,2H),8.50(dt,J＝8.8,2.5Hz,1H),8.20(d,J＝8.9Hz,1H),7.65(dd,J＝8.3,4.2Hz,1H),7.45(d,J＝2.1Hz,2H),5.3 7(s,2H),4.48(d,J＝12.8Hz,1H),3.70(d,J＝8.4Hz,1H),3.65-3.42(m,4H),3.10(ddd,J＝30.2,13 .8,6.7Hz,3H),2.46(s,3H),1.69(d,J＝7.6Hz,1H),1.40(q,J＝7.5Hz,2H),1.27(d,J＝7.0Hz,3H).

Example 59: Synthesis of Compound A159

Referring to the synthesis of compound A160 in Example 58, phenylboronic acid was used instead of 6-quinolineboronic acid pinacol ester to synthesize compound A159. m/z[MH] ^- =732.4. 1H NMR (400 MHz, DMSO-d ₆ )δ10.41(s,1H),8.58(s,1H),8.14(dd,J＝6.7,3.0Hz,2H),7.56(dd,J＝5.1,2.0Hz,3H),7.44(s,2H),5.32(s,2H),4.46(d,J＝11.9Hz,1H),3.67(t ,J＝8.8Hz,1H),3.48(s,2H),3.27(d,J＝12.3Hz,2H),3.07(dt,J＝29.6,6 .6Hz,3H),2.46(s,3H),1.67(d,J＝7.8Hz,1H),1.39(s,2H),1.26(s,3H).

Example 61: Synthesis of Compound A162

Referring to the synthesis of compound A26 in Example 15, 6-(4-methylpiperazine-1-yl)pyridine-3-boronic acid pinacol ester was used instead of A26-1 to synthesize compound A162. m/z[MH] ^- =793.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.36(s,1H),8.79(d,J＝2.4Hz,1H),8.57(s,1H),8.19-8.08(m,1H),7.40(s,2H),6.96(d,J＝9.1Hz,1H),5.26(s,2H),4.53(d,J＝12.3Hz,1H),3.60(t,J＝5.1Hz,4H),3.5 2(s,2H),3.26–3.23(m,2H),2.99(t,J＝8.6Hz,3H),2.82(d,J＝11.1Hz,1H),2.64(d,J ＝11.0Hz,1H),2.44(s,3H),2.41(t,J＝5.0Hz,4H),2.23(s,3H),1.21(t,J＝7.5Hz,3H).

Example 62: Synthesis of Compound A163

Referring to the synthesis of compound A26 in Example 15, A145-1 was used instead of A36-1, and pyrazolo[1,5-a]pyridine-5-boronic acid pinacol ester was used instead of A26-1 to synthesize compound A163. m/z[MH] ^- ＝746.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.40(s,1H),8.84(d,J＝7.4Hz,1H),8.60(s,1H),8.46(d,J＝1.8Hz,1H),8.12 (d,J＝2.3Hz,1H),7.54(dd,J＝7.3,1.9Hz,1H),7.45(s,2H),6.87(d,J＝2.2Hz,1H) ,5.34(s,2H),4.57(d,J＝12.5Hz,1H),3.56(d,J＝11.9Hz,4H),3.12–3.01(m,3H) ,2.88(d,J＝11.3Hz,1H),2.70(d,J＝10.4Hz,1H),2.48(s,3H),1.28–1.24(m,3H).

Example 63: Synthesis of Compound A165

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

Example 64: Synthesis of Compound A166

Referring to the synthesis of compound A26 in Example 15, A145-1 was used instead of A36-1, and p-fluorophenylboronic acid was used instead of A26-1 to obtain compound A166. ESI (m/z) = 724.4 [MH] ^- . ¹ H NMR (400MHz, DMSO-d ₆ )δ10.36(s,1H),8.57(s,1H),8.24–8.10(m,2H),7.43–7.33(m,4H),5.29(s,2H),4.53(d,J＝12.5Hz,1H),3.57–3.47(m,3H ),3.24(s,1H),3.01(p,J＝8.6,7.8Hz,3H),2.84(d,J＝11.2Hz,1H),2.69–2.62(m,1H),2.44(s,3H),1.22(t,J＝7.4Hz,3H).

Example 65: Synthesis of Compound A167

Referring to the synthesis of compound A26 in Example 15, A145-1 was used instead of A36-1, and 2-methylpyridine-4-boronic acid was used instead of A26-1 to obtain compound A167. m/z[M+H] ⁺ =723.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.41(s,1H),8.65(d,J＝5.1Hz,1H),8.60(s,1H),7.93(s,1H),7.85(d,J＝5.1Hz,1H),7.44(s,2H),5.33(s,2H),4.56(d,J＝12.5Hz,1H),3.5 5(s,3H),3.30(s,2H),3.06(d,J＝8.3Hz,2H),2.87(d,J＝11.3Hz,1H),2.70(d,J＝10.7Hz,1H),2.60(s,3H),2.47(s,3H),1.26(d,J＝7.2Hz,3H).

Example 66: Synthesis of Compound A168

Referring to the synthesis of compound A26 in Example 15, A145-1 was used instead of A36-1, and 1-methylpyrazole-4-boronic acid pinacol ester was used instead of A26-1 to obtain compound A168. m/z[M+H] ⁺ =712.4. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.36(s,1H),8.55(s,1H),8.30(s,1H),7.91(s,1H),7.41(s,2H),5.24(s,2H),4.52(d,J＝12.0Hz,1H),3.91(s,3H),3.50 (d,J＝10.9Hz,4H),2.99(q,J＝8.4Hz,3H),2.81(d,J＝11.2Hz,1H),2.64(d,J＝9.7Hz,1H),2.44(s,3H),1.19(d,J＝7.4Hz,3H).

Example 67: Synthesis of Compound A174

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

Example 68: Synthesis of Compound A175

Referring to the synthesis of compound A160 in Example 58, compound A175 was synthesized by replacing A20-5 with A1-4. m/z [M+H] ⁺ = 773.4. 1H NMR (400 MHz, DMSO-d ₆ ) δ 10.48 (s, 1H), 9.00 (dd, J = 4.2, 1.7 Hz, 1H), 8.79 (d, J = 1.9 Hz, 1H), 8.62-8.54 (m, 2H), 8.48 (dd, J = 8.8, 1.9 Hz, 1H), 8.18 (d, J = 8.8 Hz, 1H), 8.10 (d, J = 8.6 Hz, 1H), 8.01 (d, J = 2.1 Hz, 1H), 7.6 4(dd,J＝8.3,4.2Hz,1H),5.47(s,2H),4.48(d,J＝12.8Hz,1H),3.71(t,J＝9.2Hz,1H),3.49(d,J＝12.4Hz,3H) ,3.09(dp,J＝28.3,7.1Hz,4H),2.47(s,3H),1.70(d,J＝7.9Hz,1H),1.52–1.34(m,2H),1.28(d,J＝7.3Hz,3H).

Example 69: Synthesis of Compound A180

Referring to the synthesis of compound A26 in Example 15, A145-1 was used instead of A36-1, and pyridine-4-boronic acid was used instead of A26-1 to obtain compound A180. m/z[M+H] ⁺ =709.4. ¹ HNMR (400MHz, DMSO-d ₆ )δ10.41(s,1H),8.76(d,J＝5.1Hz,2H),8.56(s,1H),8.02(d,J＝5.0Hz,2H),7.41(s,2H),5.31(s,2H),4.53(d,J＝12.5Hz,1H),3.51(d,J =11.5Hz,3H),3.27(s,1H),3.02(p,J＝9.4,8.5Hz,3H),2.84(d,J＝11.2Hz,1H),2.66(d,J＝11.0Hz,1H),2.44(s,3H),1.24–1.20(m,3H).

Example 70: Synthesis of Compound A33

1) Synthesis of intermediate A33-3:

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

2) Synthesis of intermediate A33-4:

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

3) Synthesis of intermediate A33-5:

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

4) Synthesis of product A33:

Compound A1-7 (30 mg, 0.19 mmol), N-hydroxy-7-azabenzotriazole (29 mg, 0.22 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (45 mg, 0.23 mmol) were dissolved in N,N-dimethylformamide (0.5 mL) solution and reacted for 1 hour under stirring at 25° C. Compound A33-5 (0.1 g, 0.18 mmol) and N,N-diisopropylethylamine (70 mg, 0.54 mmol) were then added to the reaction solution in sequence, and the reaction solution was continued to react for 1 hour under stirring at 25° C. LCMS showed that the reaction was complete, and the reaction solution was added to water, extracted with dichloromethane, washed with water, washed with saturated brine, dried with sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (dichloromethane/ethyl acetate) to obtain a white product A33 (24.6 mg, yield: 19.86%, purity: 98.67%). m/z [MH] ^- = 705.3. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.34(s,1H),8.60(s,1H),8.02–7.89(m,2H),7.57–7.40(m,5H),6.89(s,1H),5.16(s,2H),4.56(d,J＝12.7Hz,1H),3.65–3.5 1(m,3H),3.25(s,1H),3.01(d,J＝8.8Hz,3H),2.85(d,J＝11.2Hz,1H),2.67(d,J＝11.1Hz,1H),2.47(s,3H),1.25(d,J＝7.4Hz,3H).

Example 71: Synthesis of Compound A179

Referring to the synthesis of compound A160 in Example 58, 4-pyridine boronic acid was used instead of 6-quinoline boronic acid pinacol ester to synthesize compound A179. m/z [M+H] ⁺ = 735.4. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.40(s,1H),8.79(d,J＝5.2Hz,2H),8.58(s,1H),8.19–7.96(m,2H),7.43(s,2H),5.32(s,2H),4.45(d,J＝12.4Hz,1H),3.68(d,J＝8. 7Hz,1H),3.48(s,2H),3.20–2.97(m,4H),2.46(s,3H),2.02(d,J＝8.6Hz,1H),1.67(d,J＝8.2Hz,1H),1.39(s,2H),1.26(d,J＝6.5Hz,3H).

Example 72: Synthesis of Compound A164

Reference Example 58 Synthesis of Compound A160 using Compound A164 was synthesized by replacing 6-quinolineboronic acid pinacol ester. m/z[M+H] ⁺ =740.3. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.37(s,1H),8.53(s,1H),7.76(d,J＝4.3Hz,2H),7.41(t,J＝5.7Hz,2H),7. 22(t,J＝4.4Hz,1H),5.25(s,2H),4.41(d,J＝11.8Hz,1H),3.64(q,J＝8.8Hz,2H) ,3.46–3.42(m,2H),3.28-2.20(m,2H),3.11–3.05(m,1H),2.99(dd,J＝14.0,7 .2Hz,1H),2.42(s,3H),1.63(q,J＝7.8Hz,1H),1.37(m,2H),1.25-1.20(m,4H).

Example 73: Synthesis of Compound A169

Reference Example 70 Synthesis of Compound A33 using replace Compound A169 was synthesized. m/z[M+H] ⁺ =672.3. ¹ H NMR (400 MHz, chloroform-d) δ11.78 (s, 1H), 8.76 (s, 1H), 8.58 (s, 1H), 7.93 (d, J=8.9 Hz, 1H), 7.02-6.94 (m, 1H), 5.64 (d, J=13.1 Hz, 1H), 5.04 (s, 2H), 4.77 (d, J=12.8 Hz, 1H), 3.8 5–3.71(m,2H),3.47(s,1H),3.15(q,J＝7.5Hz,2H),3.05(s,1H),2.85–2.67(m,2H), 2.56(s,3H),2.11(tt,J＝8.2,5.1Hz,1H),1.30(t,J＝7.5Hz,3H),1.13–1.00(m,4H).

Example 74: Synthesis of Compound A170

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

Example 75: Synthesis of Compound A176

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

Example 76: Synthesis of Compound A177

Reference Example 58 Synthesis of Compound A160 using Compound A177 was synthesized by replacing 6-quinolineboronic acid pinacol ester. m/z[M+H] ⁺ =740.4. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.36(s,1H),8.59(s,1H),7.53–7.39(m,2H),6.96–6.83(m,1H),5.24(s,2H),4.44(d,J＝11.5Hz,1H),4.30(q,J＝2.8Hz,2H),3.84(t,J＝5.4Hz,2H),3.65(t,J＝8.8Hz,2H) z,1H),3.52–3.40(m,2H),3.27(d,J＝10.4Hz,2H),3.04(dp,J＝28.6,7.1Hz,2H),2.5 9-2.50(m,2H),2.46(s,3H),1.65(d,J＝7.4Hz,1H),1.38(s,2H),1.26-1.20(m,4H).

Example 77: Synthesis of Compound A182

1) Synthesis of intermediate A182-3:

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

2) Synthesis of intermediate A182-4:

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

3) Synthesis of A182

Referring to the synthesis of compound A160 in Example 58, A182-4 was used instead of A160-3 to synthesize compound A182. m/z[M+H] ⁺ =735.3. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.38(s,1H),8.73(d,J＝4.8Hz,1H),8.58(d,J＝18.1Hz,1H),8.17(d,J＝7.9Hz,1H),7.98(td,J＝7.7,1.8Hz,1H),7.53(dd,J＝7.6,4.9Hz,1H),7.47-7.38(m,2H),5.31(s,2H),4.43(d,J＝12 .7Hz,1H),3.73–3.58(m,1H),3.54–3.37(m,3H),3.27(dd,J＝9.8,5.1Hz,1H),3.13–3.05(m,1H ),3.03–2.95(m,1H),2.43(s,3H),1.65(d,J＝7.4Hz,1H),1.48–1.29(m,2H),1.26–1.20(m,4H).

Example 78: Synthesis of Compound A181

1) Synthesis of intermediate A181-2:

Compound A181-1 (50 g, 342 mmol) was added to acetonitrile (500 mL), and then a 2 mol/L tetrahydrofuran solution of dimethylamine (200 mL) was added, and the mixture was reacted at 50 ^°C for 4 hours. Hydrazine hydrate (51.35 g, 820.6 mmol) was added, and the mixture was refluxed for overnight reaction. LCMS showed that the reaction was complete, and the reaction solution was returned to room temperature, and the mixture was concentrated under reduced pressure (remaining 230 mL). Solids were precipitated, and the filter cake was obtained by filtration. The filter cake was dried to obtain the product (39 g, yield: 89.7%). m/z[M+H] ⁺ =128.0.

2) Synthesis of A181:

Referring to the synthesis of compound A182 in Example 77, compound A181 was synthesized by using A181-2 instead of A182-1. m/z [M+H] ⁺ = 701.4. ¹ H NMR (400 MHz, CDCl ₃ ) δ 11.66 (s, 1H), 8.83 (s, 1H), 8.59 (s, 1H), 7.95 (d, J = 8.8 Hz,1H),6.98(d,J＝8.9Hz,1H),4.98(s,2H),4.85-4.70(m,0.5H),4.30-4.00(m,2H),3.95-3.55(m,1.5H),3.23(dd,J＝52.7,10.8Hz,2 H),3.11(s,6H),3.09-3.00(m,1H),2.54(s,3H),2.34(d,J＝26.2Hz,1H),1.80-1.70(m,1H),1.55-1.35(s,2H),1.28(t,J＝7.5Hz,4H).

Example 79: Synthesis of Compound A183

Reference Example 77 Synthesis of Compound A182 using replace Compound A183 was synthesized. m/z[M+H] ⁺ =709.1. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.38(s,1H),8.74–8.70(m,1H),8.58(s,1H),8.18(dt,J＝7.9,1.1Hz,1H),7.98(td,J＝7.8,1.8Hz,1H),7.53(ddd,J＝7.6,4.8,1.2Hz,1H),7.47–7.38(m,2H),5.32(s,2H), 4.54(d,J＝12.5Hz,1H),3.52(t,J＝10.3Hz,3H),3.29–3.21(m,1H),3.02(d,J＝9.6Hz,3 H), 2.85 (d, J = 11.3Hz, 1H), 2.67 (d, J = 10.4Hz, 1H), 2.45 (s, 3H), 1.22 (t, J = 7.4Hz, 3H).

Example 80: Synthesis of Compound A184

Referring to the synthesis of compound A160 in Example 58, 3-pyridine boronic acid was used instead of 6-quinoline boronic acid pinacol ester to obtain compound A184. m/z [M+H] ⁺ = 735.4. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.42 (s, 1H), 9.30 (s, 1H), 8.75 (d, J = 4.8 Hz, 1H), 8.60 (s, 1H), 8.46 (dd, J = 8.0, 2.0 Hz, 1H), 7.62 (dd, J = 7.9, 4.8 Hz, 1H), 7.45 (d, J = 4.2 Hz, 2H), 5.33 (s, 2H), 4.46 (d, J = 12.1 Hz ,1H),3.69(q,J＝11.8,9.4Hz,1H),3.49(s,4H),3.20–3.09(m,1H),3.08–2.97(m,1H),2. 47(s,3H),1.68(d,J＝8.0Hz,1H),1.40(q,J＝7.7,7.2Hz,2H),1.26(td,J＝6.8,3.6Hz,4H).

Example 81: Synthesis of Compound A187

1) Synthesis of intermediate A187-1:

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

2) Synthesis of intermediate A187-2:

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

3) Synthesis of intermediate A187-3:

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

4) Synthesis of intermediate A187-4:

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

5) Synthesis of intermediate A187-5:

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

6) Synthesis of intermediate A187-6:

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

7) Synthesis of intermediate A187-7:

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

8) Synthesis of A187:

Add the following into a 10mL single-mouth bottle: (90 mg, 0.55 mmol), N-hydroxy-7-azabenzotriazole (80 mg, 0.58 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (110 mg, 0.58 mmol) were dissolved in N,N-dimethylformamide (0.5 mL) solution and reacted at 25°C for 1 hour under stirring. A187-7 (190 mg, 0.32 mmol) and N,N-diisopropylethylamine (120 mg, 0.96 mmol) were then added to the reaction solution in sequence, and the reaction solution was stirred at 25°C for 2 hours. LCMS showed that the reaction was complete, and the reaction solution was filtered to obtain a filtrate, which was subjected to reverse phase column chromatography (formic acid 0.05% aqueous solution/acetonitrile) to obtain compound A187. m/z[M+H] ⁺ =727.3. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.29(s,2H),8.55(s,1H),7.46–7.36(m,2H),5.11(s,2H),4.40(d,J＝11.5Hz,1H),3.63(q,J＝9.0Hz,1H),3.45–3.35(m,7H),3.21(d,J＝ 10.6Hz,1H),2.95(dp,J＝21.5,7.2Hz,2H),2.43(s,3H),1.92–1.87(m,4H),1.60(q,J＝7.6Hz,1H),1.40–1.28(m,2H),1.17(t,J＝7.4Hz,4H).

Example 82: Synthesis of Compound A185

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

Example 83: Synthesis of Compound A186

Reference Example 70 Synthesis of Compound A33 using Compound A186 was synthesized by using A187-2 instead of A33-2. m/z [M+H] ⁺ = 743.1. ¹ H NMR (400MHz, DMSO_d ₆ ): δ10.34(s,1H),8.57(s,1H),7.54–7.37(m,2H),5.13(s,2H),4.42(d,J＝11.7Hz,1H),3.70(t,J＝4.7Hz,4H),3.64(d,J＝8.0Hz ,2H),3.48-3.38(m,5H),3.23(d,J＝11.4Hz,1H),3.09–2.87(m,2H),2.45(s,3H),2.12–1.89(m,1H),1.62(d,J＝7.0Hz,1H),1.38 -1.30(m,2H),1.21-1.13(m,4H).

Example 84: Synthesis of Compound A188

Referring to the synthesis of compound A187 in Example 81, (S)-2-methylpyrrolidine was used instead of tetrahydropyrrole to synthesize compound A188. m/z[MH] ^- =739.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.30(s,1H),8.55(s,1H),7.48–7.36(m,2H),5.10(s,2H),4.40(d,J＝11. 4Hz,1H),3.92(td,J＝6.7,3.5Hz,1H),3.62(d,J＝8.2Hz,1H),3.44(dd,J＝8.8 ,4.1Hz,4H),3.21(d,J＝11.0Hz,1H),2.96(q,J＝10.4,8.5Hz,2H),2.42(s,3H ),2.10–1.77(m,4H),1.61(d,J＝8.6Hz,2H),1.33(s,2H),1.24–1.13(m,7H).

Example 85: Synthesis of Compound A189

Referring to the synthesis of compound A187 in Example 81, (R)-2-methylpyrrolidine was used instead of tetrahydropyrrole to synthesize compound A189. m/z[MH] ^- =739.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.30(s,1H),8.55(s,1H),7.45–7.35(m,2H),5.10(s,2H),4.40(d,J＝11.3H z,1H),3.92(dq,J＝7.0,3.7Hz,1H),3.68–3.58(m,1H),3.47–3.35(m,4H),3.20 (d,J＝10.2Hz,1H),2.95(dp,J＝28.2,7.0Hz,2H),2.43(s,3H),2.13–1.78(m,4H ),1.61(dq,J＝10.6,6.3,5.6Hz,2H),1.33(t,J＝6.7Hz,2H),1.22–1.13(m,7H).

Example 86: Synthesis of Compound A190

Reference Example 81 Synthesis of Compound A187 using Compound A190 was synthesized by replacing tetrahydropyrrole. m/z [M+H] ⁺ = 782.4. ¹ H NMR (400 MHz, chloroform-d) δ 8.85 (s, 1H), 8.59 (s, 1H), 7.99 (d, J = 8.9 Hz, 1H), 6.99 (d, J = 8.9 Hz, 1H), 4.97 (s, 2H), 4.84–3.77 (m, 3H), 3.59 (t, J = 5.1 Hz, 5H), 3.39–3.01 (m, 3H), 2.70 (t, J = 5.0 Hz, 4H), 2.54 (s, 3H), 2.41–1.76 (m, 2H), 1.27 (dd, J = 13.7, 6.1 Hz, 7H), 0.48 (dd, J = 9.3, 4.9 Hz, 4H).

Example 87: Synthesis of Compound A191

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

Example 88: Synthesis of Compound A192

Reference Example 81 Synthesis of Compound A187 using Compound A192 was synthesized by replacing tetrahydropyrrole. m/z [M+H] ⁺ = 715.4. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.31(s,1H),8.52(s,1H),7.47–7.33(m,2H),5.09(s,2H),4.40(d,J＝11.1Hz,1H),3.62(d,J＝8.5Hz,1H),3.43(p,J＝6.2,5.3Hz,4H),3. 21(d,J＝9.0Hz,2H),2.94(s,5H),2.42(s,3H),1.60(d,J＝6.2Hz,1H),1.28(d,J＝40.1Hz,2H),1.17(t,J＝7.4Hz,4H),1.07(t,J＝7.0Hz,3H).

Example 89: Synthesis of Compound A193

Reference Example 89 Synthesis of Compound A187, Compound A193 was synthesized by replacing tetrahydropyrrole with azetidine. m/z[MH] ^- =711.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.30(d, J=14.4 Hz, 2H),8.55(s, 1H),7.41(d, J=3.6 Hz, 2H),5.10(s, 2H),4.40(d, J=11.2 Hz, 1H),3.96(t, J=7.5 Hz, 5H),3.61(d, J=9.0 Hz, 1H),3.41(d, J=9.5 Hz, 1H) ,3.20(d,J＝11.0Hz,1H),3.02–2.95(m,1H),2.94–2.85(m,1H),2.43(s,3H),2.36–2 .30(m,2H),1.60(d,J＝6.6Hz,1H),1.33(s,2H),1.23(s,1H),1.16(t,J＝7.4Hz,4H).

Example 90: Synthesis of Compound A194

Reference Example 81 Synthesis of Compound A187, Compound A194 was synthesized by replacing tetrahydropyrrole with 3-methoxy-azetidine. m/z[MH] ^- =741.4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.28(s,1H),8.55(s,1H),7.45-7.36(m,2H),5.10(s,2H),4.40(d,J＝11.5Hz,1H),4.31(tt,J＝6.1,4.3Hz,1H),4.17(dd,J＝8.8,6.2Hz,2H),3.79(dd,J＝9.0,4.2Hz,2H),3.65-3.5 6(m,1H),3.41(d,J＝8.9Hz,2H),3.22(s,3H),3.19(s,1H),3.04–2.86(m,2H),2.42(s,3H), 1.60(d,J＝6.8Hz,1H), 1.33(d,J＝6.7Hz,2H), 1.22(d,J＝5.3Hz,1H), 1.16(t,J＝7.4Hz,4H).

Example 91: Synthesis of Compound A195

Reference Example 81 Synthesis of Compound A187 using Compound A195 was synthesized by replacing tetrahydropyrrole. m/z [M+H] ⁺ = 749.0. ¹ H NMR (400MHz, CDCl ₃ ): δ8.53(s,1H),8.48(s,1H),7.90(d,J＝8.8Hz,1H),6.93(d,J＝8.8Hz,1H),5.44-4.92(m,2H),4.64-4.70(m,1H),4.44-4.38(m,4H),4.14-3 .97(m,2H),3.79-3.61(m,2H),3.26-2.98(m,2H),2.47(s,3H),2.29- 2.26(m,1H),2.15-2.10(m,1H),1.98-1.71(m,2H),1.24-1.18(m,4H).

Example 92: Synthesis of Compound A196

Reference Example 81 Synthesis of Compound A187 using Compound A196 was synthesized by replacing tetrahydropyrrole. m/z [M+H] ⁺ = 757.0. ¹ H NMR (400MHz, DMSO_d ₆ ): δ10.32(s,1H),8.50(brs,1H),7.42(q,J＝8.4Hz,2H),5.11(s,2H),4. 44-4.38(m,1H),4.06-4.01(m,1H),3.65-3.58(m,1H),3.49-3.37(m,5H) ,3.30-3.18(m,3H),3.23(s,3H),3.01-2.88(m,2H),2.41(s,3H),2.06-1 .95(m,2H),1.62-1.57(m,1H),1.37-1.29(m,2H),1.17(t,J=7.2Hz,4H).

Example 93: Synthesis of Compound A197

Reference Example 81 Synthesis of Compound A187 using Compound A197 was synthesized by replacing tetrahydropyrrole. m/z [M+H] ⁺ = 757.0. ¹ H NMR (400MHz, DMSO_d ₆ ): δ10.28(s,2H),8.51(s,1H),7.43-7.37(m,2H),5.10(s,2H),4.39(d,J＝11.6Hz,1H),4.04-4.02(m,1H),3.65-3.58(m,1H),3.48-3.37( m,5H),3.33-3.18(m,6H),3.01-2.88(m,2H),2.42(s,3H),2.03-1.98(m,2H),1.61-1.56(m,1H),1.34-1.31(m,2H),1.16(d,J＝7.6Hz,4H).

Example 94: Synthesis of Compound A198

Reference Example 81 Synthesis of Compound A187 using Compound A198 was synthesized by replacing tetrahydropyrrole. m/z [M+H] ⁺ = 769.1. ¹ H NMR (400MHz, CDCl ₃ ): δ8.79(s,1H),8.52(s,1H),7.89(d,J＝9.2Hz,1H),6.91(d,J＝9.2Hz,1 H),4.88(s,2H),4.68(s,1H),4.14-4.12(m,3H),4.02-3.96(m,3H),3.8 2-3.79(m,3H),3.62-3.58(m,1H),3.25-2.99(m,4H),2.47(s,3H),2.26 -2.23(m,1H),2.10-2.07(m,2H),1.97-1.52(m,4H),1.23-1.18(m,3H).

Example 95: Synthesis of Compound A201

Reference Example 59 Synthesis of Compound A159 using replace Compound A201 was synthesized. m/z [M+H] ⁺ = 734.3. ¹ H NMR (400MHz, DMSO_d ₆ ): δ10.38(d,J＝12.0Hz,2H),8.59(s,1H),8.14(dd,J＝6.7,3.0Hz,2H),7.56(dd,J＝5.1,2.0Hz,3H),7.43(s,2H),5.31(s,2H),4.47(s,1H), 3.69(d,J＝8.6Hz,1H),3.47(d,J＝7.4Hz,2H),3.29-2.94(m,4H),2.46(s,3H),1.67(d,J＝7.7Hz,1H),1.40-1.33(m,2H),1.29-1.23(m,4H).

Example 96: Synthesis of Compound A202

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

Example 97: Synthesis of Compound A199

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

Example 98: Synthesis of Compound A200

Reference Example 58 Synthesis of Compound A160 using Replace 6-quinolineboronic acid pinacol ester and use Compound A200 was synthesized instead of A1-7. m/z[M+H] ⁺ =724.4. ¹ H NMR (400 MHz, DMSO_d ₆ )δ10.31(s,1H),9.80(s,1H),7.40(d,J＝2.7Hz,2H),7.26-7.14(m,2H),6.83(q,J＝7.4Hz,3H),5.20(s,2H),4.26(q,J＝2.8Hz,3H),3.81(t,J＝5.4Hz,2H),3.58(d,J＝8.6Hz,1H),3.38(d,J＝ 12.6Hz,3H),3.27(d,J＝13.2Hz,3H),3.15(d,J＝11.6Hz,1H),3.05(dd,J＝13.8,7.2Hz,1H),2. 99–2.90(m,1H),1.66–1.57(m,1H),1.34(dd,J＝11.9,4.6Hz,1H),1.21(q,J＝7.5,6.5Hz,4H).

Biological testing section

Experimental Example 1 Biological Activity Test

ATPase activity assay of WRN helicase

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

45 nt oligo DNA single strand FLAP26 (TTTTTTTTTTTTTTTTTTTTTTCCAAGTAAAACGACGGCCAGTGC) was synthesized by Ascent Inc. See, for example, Brosh RM Jr et al., J Biol Chem, 2002 Jun; 277(26):23236-45. Prepare reaction buffer (30mM Tris pH 7.5, 2mM MgCl ₂ , 0.02% BSA, 50mM NaCl, 0.1% pluronic F127), add 5μL 3× test compound (diluted to 0.5% DMSO in reaction buffer, final starting concentration is 10μM, 1:3 dilution, 9 gradients), 5μL 3× WRN recombinant protein and 3× ATP substrate solution (diluted in reaction buffer, final concentrations of WRN and ATP are 10nM and 300μM, respectively) to a 384-well transparent plate, shake and mix, and incubate at 37°C for 3 hours. Next, prepare 3× FLAP26 solution with reaction buffer, take 5μL solution (final concentration of FLAP26 is 0.4nM) and add it to a 384-well transparent plate, shake and mix, start the enzymatic reaction, and incubate at room temperature for 30 minutes.

Take 5 μL of the above mixture and transfer it 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 ₅₀ value of the test compound.

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

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.

Tumor cell proliferation inhibitory activity assay

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.

Human colon adenocarcinoma cell SW48 cells were cultured in vitro in monolayers, and the culture conditions were: DMEM medium containing 10% fetal bovine serum, 1% penicillin and streptomycin, 37°C 5% CO _2. Human colon adenocarcinoma cell HCT116 cells were cultured in vitro in monolayers, and the culture conditions were: McCoy's5A medium containing 10% fetal bovine serum, 1% penicillin and streptomycin, 37°C 5% CO _2. DLD1-WRN-KO cells were cultured in vitro in monolayers, and the culture conditions were: 1640 medium containing 10% fetal bovine serum, 1% penicillin and streptomycin, 37°C 5% CO _2. Trypsin digestion and passage were performed twice a week. After the test compound was treated with microsatellite unstable SW48 cell line and HCT116 cell line and control cell DLD1-WRN-KO cell line for 4 days, the ATP level was detected using the CellCounting-Lite kit of Novozyme to evaluate the inhibitory effect of the test compound on the growth of tumor cell lines.

In the present application, SW48 cell line, HCT116 cell line and DLD1-WRN-KO cell line were inoculated in 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, 1:3 times dilution of 9 gradients, and a DMSO treatment group was set up. Cultured in a 37 ° C / 5% CO ₂ incubator for 4 days. To test the proliferation inhibition of tumor cells by the test compound, the cells were balanced at room temperature for 30 minutes, and then 100 μL of cell proliferation detection reagent CellCounting-Lite (CCL) was added to each well, and incubated in the dark for 10 minutes after oscillation for 5 minutes. The chemiluminescence value was read using the Thermo Varioskan LUX-3020 multifunctional microplate reader to convert it into a proliferation index to calculate the inhibition rate of tumor cell proliferation by the compound, and the inhibition rate value and the logarithm of the compound concentration were fitted using nonlinear regression (dose response-variable slope) to obtain the IC ₅₀ value of the compound.

Table 2 Tumor cell proliferation inhibition activity data of the compounds in the examples of the present application

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

Experimental Example 2 Liver microsome stability test

The first-phase metabolic stability of the test compounds was assessed in liver microsomes of CD-1 mice, Sprague-Dawley rats, beagle dogs, cynomolgus monkeys and humans.

Experimental system:

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.

Experimental introduction:

The test sample and the control compound were incubated with animal and human liver microsomes at 37±1°C for a certain period of time, with the longest incubation time being 60 minutes. Samples were taken out at the specified time point and the reaction was terminated with acetonitrile or other organic solvents containing internal standards. After centrifugation, the resulting supernatant was detected by liquid chromatography-tandem mass spectrometry (LC-MS/MS).

Experimental methods:

1. Preparation of buffer

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

2. Preparation of working solution

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.

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

3. Preparation of Liver Microsome Solution

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

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

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

5. Preparation of Stop Solution

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

6. Incubation process

Incubations will be done in 96-well plates. Prepare 8 incubation plates, named T0, T5, T15, T30, T45, T60, Blank60, and NCF60. The first 6 plates correspond to reaction time points of 0, 5, 15, 30, 45, and 60 minutes, respectively. No test or control compound is added to the Blank60 plate, and samples are taken after 60 minutes of incubation. In the NCF60 plate, potassium phosphate buffer is used instead of NADPH regeneration system solution for incubation for 60 minutes. All condition samples are three parallels.

Mix the microsomes with the test or control compound, and then place the incubation plates Blank60, T5, T15, T30, T45 and T60, except T0 and NCF60, in a 37°C water bath for pre-incubation for about 10 minutes. Add the stop solution to the incubation plate T0 first, then add the NADPH regeneration system working solution, and add 98μL potassium phosphate buffer to each sample well of the incubation plate NCF60 to start the reaction. After the pre-incubation of the incubation plates Blank60, T5, T15, T30, T45 and T60, add 98μL NADPH regeneration system working solution to each sample well to start the reaction. The reaction temperature is 37±1°C, the final reaction volume is 200μL, and the reaction system includes 0.5mg/mL microsomes, 1.0μM substrate, 1mM NADP, 6mM ISO and 1unit/mL IDH.

At 5, 15, 30, 45 and 60 minutes, cold stop solution containing internal standard was added to the reaction plate to stop the reaction.

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

Sample analysis

Sample analysis was performed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) without standard curves and quality control samples. Semi-quantitative determination was performed using the ratio of the analyte peak area to the internal standard peak area. Retention times of analytes and internal standards, chromatogram acquisition, and chromatogram integration were processed using the software Analyst (Sciex, Framingham, Massachusetts, USA).

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

Data analysis

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

CL _int(mic) = 0.693/T _1/2 /microsomal protein content (microsomal concentration during incubation mg/mL)

CL _int(liver) = CL _int(mic) × amount of microsomal protein in liver (mg/g) × liver weight to body weight ratio

According to the well stir model, the hepatic intrinsic clearance and hepatic clearance can be converted by the following formula.

CL _(liver) = (CL _{int (liver)} *Q _h )/(CL _{int (liver)} +Q _h )

The parameters in the formula are shown in the table below.

Parameters in Data Analysis Formulas

Experimental Example 3 Hepatocyte Metabolic Stability Test

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

Prepare 0.5x10 ⁶ /mL hepatocyte suspension with preheated culture medium, then add 198 μL preheated cell suspension to 96-well plate. Add 2 μL of the test compound to each well of the 96-well plate to make a final concentration of 1 μM, and set 2 replicates. For samples with T=0 minutes, mix the compound and cells thoroughly for 1 minute, then immediately take 25 μL of the sample and add it to 125 μL of stop solution (acetonitrile solution containing 200 ng/mL tolbutamide and 200 ng/mL labetalol) in an ice bath and mix. At the same time, place all plates in an incubator at 37°C and 5% CO ₂ , and set the shaker to 600 rpm. The samples were mixed at 15, 30, 60, and 90 minutes of incubation, and 25 μL of the sample was added to 125 μL of stop solution (containing 200 ng/mL tolbutamide and 200 ng/mL labetalol in acetonitrile) in an ice bath, mixed, and shaken at 500 rpm for 10 minutes. Subsequently, centrifuged at 3220 × g for 20 minutes at 4 °C. 80 μL of supernatant was taken from each well and transferred to another 96-well plate containing 240 μL of ultrapure water. The intrinsic clearance (CLint) and half-life (T1/2) were then analyzed and calculated using LC-MS/MS.

Experimental Example 4 Pharmacokinetic Test in Rats

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

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 jugular vein blood collection. 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, snap-frozen in dry ice, and then stored in an ultra-low temperature freezer at -60°C or lower until LC-MS/MS analysis.

Plasma concentrations were determined using LC-MS/MS. WinNonlin Version 6.3 (Pharsight, Mountain View, CA) pharmacokinetic software was used to process the plasma drug concentration data of the compound using a non-compartmental model. The linear-log trapezoidal method was used to calculate the relevant pharmacokinetic parameters.

Experimental Example 5: Pharmacokinetic test in mice

In this experiment, the pharmacokinetic behavior of the tested compounds was investigated in BALB/c mice after intravenous (IV) and oral (PO) administration.

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

Plasma concentrations were determined using LC-MS/MS. WinNonlin Version 6.3 (Pharsight, Mountain View, CA) pharmacokinetic software was used to process the plasma drug concentration data of the compound using a non-compartmental model. The linear-log trapezoidal method was used to calculate the relevant pharmacokinetic parameters.

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

Comparative compound B was prepared by referring to the synthesis method of compound A20, except that A1-4 was used instead of A20-5. ESI(m/z)＝696.3[M+H] ⁺ .1H NMR(400MHz,DMSO-d ₆ )δ8.35(s,2H),8.08(dt,J＝5.4,3.2Hz,2H),8.01(d,J＝8.6Hz,1H),7.95(t,J＝1.3Hz,1H),7.72–7.66(m,1H),7.54–7.48(m,3H),5.37 (s,2H),4.53(d,J＝11.6Hz,1H),3.58–3.22(m,4H),3.00(d,J＝7.6Hz,3H),2.84(d,J＝11.0Hz,1H),2.38(s,3H),1.19(t,J＝7.3Hz,3H).

Experimental Example 6 hERG inhibition test

HEK293 cells were cultured in DMEM medium containing 10% fetal bovine serum and 0.8mg/mL G418 at 37℃ and 5% _CO2 . The cells were digested with TrypLE ^TM Express and centrifuged to adjust the cell density to 2×10 ⁶ cells/mL. The cells were then gently mixed on a room temperature balanced shaker for 15-20min and then put on the machine for patch clamp detection. The culture medium of the prepared cells was replaced with extracellular fluid. The intracellular and extracellular fluids were drawn 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 recorded the voltage stimulation of the whole-cell hERG potassium current, and the experimental data were 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 solution without the compound for each cell was used as its own control group. At least two cells were used for each concentration and the detection was repeated twice independently. All electrophysiological experiments were performed at room temperature.

Data analysis: First, the current after each drug concentration was normalized with the blank control current. Then calculate the inhibition rate corresponding to each drug concentration For each concentration the mean and standard error were calculated and the half-inhibitory concentration for each compound was calculated: 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, _IC50 represents the half-inhibitory concentration, and HillSlope represents the Hill coefficient. Curve fitting and _IC50 calculation were completed using Graphpad software.

Experimental Example 7 Cytochrome oxidase P450 inhibition test

1) Preparation of buffer:

100mM K-Buffer: Mix 9.5mL stock solution A with 40.5mL stock solution B and adjust the total volume to 500mL with ultrapure water. mL, titrate the buffer to pH 7.4 with KOH or H ₃ PO ₄ .

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

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

2) Preparation of test substances

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.

3) In vitro incubation

The in vitro incubation system of liver microsomes for the study of CYP450 enzyme metabolic phenotype 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.

4) Detection of parent drug or metabolites

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

Experimental Example 8 Mouse Tumor Pharmacodynamic Model

In this experimental example, the in vivo efficacy of the tested compounds in a mouse xenograft tumor model was evaluated after intragastric administration (PO).

Human colon adenocarcinoma cell SW48 cells were cultured in monolayer in vitro, and the culture conditions were: DMEM medium containing 10% fetal bovine serum, 1% penicillin and streptomycin, and cultured at 37°C 5% CO _2. Human colon adenocarcinoma cell HCT116 cells were cultured in monolayer in vitro, and the culture conditions were: McCoy's5A medium containing 10% fetal bovine serum, 1% penicillin and streptomycin, and cultured at 37°C 5% CO _2. Digestion and passage were performed with trypsin twice a week. When the cell saturation was 80%-90%, the cells were collected, counted, and inoculated. 0.1mL (10 ⁷ ) SW48 cells or 0.1mL (5*10 ⁶ ) HCT116 cells were subcutaneously inoculated into the right back of each mouse. On the 14th day after cell inoculation, the average tumor volume reached about 150-200mm ³ , and the mice were randomly divided into groups for drug administration, and the drug was administered orally once a day, and the changes in body weight and tumor volume were recorded. After a certain number of days of drug administration, the experiment was terminated. The changes in tumor volume were counted and analyzed. The tumor growth inhibition rate (TGI%) and p value were calculated based on the data on the last day, see the table below.

Tumor volume calculation formula:

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

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

Remark:

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

b. The p value was obtained by comparing the control compound administration group and the present compound administration group using the One-way ANOVA method. Statistical analysis of p>0.05 indicates that there is no significant difference between the present compound administration group and the control compound administration group. Statistical analysis of p≤0.05 indicates that there is a significant difference between the present compound administration group and the control compound administration group. Statistical analysis of p≤0.01 indicates that there is a very significant difference between the present compound administration group and the control compound administration group.

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

In addition to those described herein, various modifications of the present invention will be apparent to those skilled in the art based on 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 (20)

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):
in: Selected from wherein R ¹ is connected to the position marked with “*” in the above group, R ² is connected to the position marked with “**” in the above group, R ³ is connected to the position marked with “***” in the above group, and R ⁴ is connected to the position marked with “****” in the above group; R ¹ is selected from -NR ¹¹ R ¹² , ^R2 is ^R4 is R ³ , R ¹¹ , R ¹² , R ¹³ , R ²¹ , R ²² , R ²³ and R ²⁴ are each independently selected at each occurrence from H, halogen, -OH, -NH ₂ , -CN, -NO ₂ , -SF ₅ , 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 heterocyclyl, 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) ₂ R ^a , -S(═O) ₂ NR ^a R ^{b -NRaRb} , -C(=O) ^NRaRb , -NRa ^- C(=O) ^Rb , -NRa ^- C(=O) ^ORb , -NRa ^- S(=O) ₂ - ^Rb , ^{-NRa - C} (=O ⁾ -NRaRb, _-P ⁽ ₌ O ⁾ _RaRb , -C1-6alkylene _- ^Ra , -C1-6alkylene-ORa ^, _{-C1-6alkylene} - ^NRaRb , -OC1-6alkylene-NRaRb, ( _{-C3-6cycloalkylene} ) ^{-CN and} ( ^{-C3-6cycloalkylene} ) ^{-C1-6haloalkyl} _; R ¹⁴ at each occurrence is independently selected from -L ¹ -(C _3-6 cycloalkyl), -L ¹ -(3-10 membered heterocyclyl), -L ¹ -(C _6-10 aryl) and -L ¹ -(5-14 membered heteroaryl); R ⁴¹ is selected from a C _3-6 hydrocarbon ring, a 3-10 membered heterocyclic ring, a C _6-10 aromatic ring, and a 5-14 membered heteroaromatic ring; Ring C, Ring D, Ring E, Ring X and Ring Z are each independently selected from a C _3-6 hydrocarbon ring, a 3-10 membered heterocyclic ring, a C _6-10 aromatic ring and a 5-14 membered heteroaromatic ring; Ring Y is absent or is selected from a C _3-6 hydrocarbon ring, a 3-10 membered heterocyclic ring, a C _6-10 aromatic ring and a 5-14 membered heteroaromatic ring; when Ring Y is absent, R ²⁴ is also absent; L ¹ is selected from a direct bond, -O-, -C(=O)-, -NRC(=O)-, -S-, -S(=O)-, -S(=O) ₂ -, C _1-6 alkylene, and -O-(C _1-6 alkylene)-; ^L2 is selected from -O-, -C(=O)-, -NRC(=O)-, -S-, -S(=O)-, -S(=O) _2- , _C1-6 alkylene and -O-( _C1-6 alkylene)-; R, ^Ra and ^Rb are each independently selected at each occurrence from H, _C1-6 alkyl, _C3-10 cycloalkyl, 3-10 membered heterocyclyl, _C6-10 aryl, 5-14 membered heteroaryl and _C6-12 aralkyl; The above alkylene, alkyl, alkenyl, alkynyl, cycloalkylene, cycloalkyl, hydrocarbon ring, heterocyclyl, heterocycle, aryl, aromatic ring, heteroaryl, heteroaromatic ring and aralkyl are each optionally substituted at each occurrence by one or more substituents independently selected from the following: halogen, -OH, =O, = _CF2 , _-NH2 , -CN, _-NO2 , _C1-6 alkyl, deuterated _C1-6 alkyl, C3-6 cycloalkyl, _3-10 membered heterocyclyl, _C6-10 aryl, 5-14 membered heteroaryl, _C6-12 aralkyl, -C(=O) ^Rc , -OC(=O) ^Rc , -C(=O) ^ORc , ^-ORc , -SRc, -S(=O ^{)Rc ,} -S(= ^O ) _2Rc ^, -S(=O ⁾ _2NRcRd ^{, -NRcRd} -C(═O ^{) NRcRd} , -NRc- ^C (═O) ^Rd , -NRc- ^C (═O) ^ORd , -NRc ^- S(═O) ₂ - ^Rd , -NRc ^{-C (} ═O) ^-NRcRd , _{-C1-6alkylene} - ^ORc , _{-C1-6alkylene} - ^NRcRd and -OC1-6alkylene- ^NRcRd , said alkylene, alkyl, cycloalkyl, heterocyclyl, aryl, _heteroaryl and aralkyl being 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 , _C1-6alkyl , _{C1-6haloalkyl} , C3-6cycloalkyl, _3-10 membered heterocyclyl, C _6-10- membered aryl, 5-14-membered heteroaryl, C _6-12 aralkyl, -OC _1-6 alkyl, and -C _1-6 alkylene-OC _1-6 alkyl; R ^c and R ^d are each independently selected at each occurrence from H, C _1-6 alkyl, C _3-10 cycloalkyl, 3-10 membered heterocyclyl, C _6-10 aryl, 5-14 membered heteroaryl and C _6-12 aralkyl, said alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and aralkyl further optionally substituted with one or more substituents independently selected from halogen, -OH, =O, -C(=O)O-tert-butyl, -NH ₂ , -CN, -NO ₂ , C _1-6 alkyl, C _1-6 haloalkyl, C _3-6 cycloalkyl, 3-10 membered heterocyclyl _, C _6-10 aryl, 5-14 membered heteroaryl, C _6-12 aralkyl and -C _1-6 alkylene-OC _1-6 alkyl; n, p and q are each independently an integer selected from 1, 2 or 3; and m is an integer of 0 or 1. The compound of claim 1 or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, wherein the compound has the structure of the following formula:

The compound of claim 1 or 2, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, wherein: ^R1 is R ¹³ is H, halogen or C _1-6 alkyl; R ¹⁴ is selected from 3-10 membered heterocyclyl, -(C _1-6 alkylene)-(3-10 membered heterocyclyl) and -O-(C _1-6 alkylene)-(3-10 membered heterocyclyl), said heterocyclyl being optionally substituted with one or more substituents independently selected from the group consisting of C _1-6 alkyl, C _3-6 cycloalkyl, 3-10 membered heterocyclyl, -C _1-6 alkylene-C _3-6 cycloalkyl and -C _1-6 alkylene-CN; and n is an integer of 1 or 2; Preferably, ^R1 is The compound of claim 1 or 2, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, wherein: ^R1 is Ring C is a benzene ring, a 5-6 membered heterocyclic ring or a 5-6 membered heteroaromatic ring; Ring D is a benzene ring or a 5-6 membered heteroaromatic ring; The above-mentioned benzene ring, heterocyclic ring and heteroaromatic ring are each optionally substituted by one or more substituents independently selected from the following: halogen, -OH, -NH ₂ , -CN, -NO ₂ , C _1-6 alkyl, deuterated C _1-6 alkyl, C _1-6 haloalkyl, -OC _1-6 alkyl, C _3-6 cycloalkyl and 3-10 membered heterocyclyl; preferably, the above-mentioned benzene ring, heterocyclic ring and heteroaromatic ring are each optionally substituted by one or more substituents independently selected from the following: halogen, C _1-6 alkyl, C _1-6 haloalkyl, -OC _1-6 alkyl and C _3-6 cycloalkyl; Preferably, ^R1 is The compound of claim 1 or 2, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, wherein R ¹ is -NR ¹¹ R ¹² ; R ¹¹ and R ¹² are each independently selected from H, C _1-6 alkyl, C _3-6 cycloalkyl, 3-10 membered heterocyclyl, C _6-10 aryl, 5-14 membered heteroaryl, C _6-12 aralkyl, -C _1-6 alkylene-R ^a and -C _1-6 alkylene-NR ^a R ^b ; ^Ra and ^Rb are each independently selected at each occurrence from H, _C1-6 alkyl, _C3-10 cycloalkyl, 3-10 membered heterocyclyl, _C6-10 aryl, 5-14 membered heteroaryl and _C6-12 aralkyl; The alkylene, alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and aralkyl groups are each optionally substituted at each occurrence with one or more substituents independently selected from the group consisting of halogen and C _1-6 alkyl; Preferably, ^R1 is The compound of claim 1 or 2, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, wherein: ^R1 is Ring E is a C _3-6 hydrocarbon ring, a 3-10 membered heterocyclic ring, a C _6-10 aromatic ring or a 5-14 membered heteroaromatic ring, wherein the hydrocarbon ring and the heterocyclic ring are optionally substituted by =CF ₂ ; R ¹³ is H, halogen, C _1-6 alkyl, C _3-6 cycloalkyl, 3-10 membered heterocyclyl, -C(=O)-(C _1-6 alkyl), -O-(C _1-6 alkyl) or -N(C _1-6 alkyl) ₂ , wherein the alkyl, cycloalkyl and heterocyclyl are further optionally substituted by C _1-6 alkyl, C _3-6 cycloalkyl or 3-10 membered heterocyclyl; n is an integer of 1 or 2; Preferably, ^R1 is The compound of any one of claims 1 to 6 or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, wherein when Y is absent, then a) Ring X is a 5-6 membered heterocyclic ring (eg, a pyridone ring) or a 5 membered heteroaromatic ring; or b). Ring X is a benzene ring, and at least one R ²³ is (—C _3-6 cycloalkylene)-CN or (—C _3-6 cycloalkylene)-C _1-6 haloalkyl. The compound of any one of claims 1 to 6, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, wherein ring X is a benzene ring and ring Y is a 5-6 membered heterocyclic ring or a 5-6 membered heteroaromatic ring. A compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein R ²¹ , R ²² , R ²³ and R ²⁴ are each independently selected from H, halogen, -SF ₅ , C _1-6 alkyl, halogenated C _1-6 alkyl, C _3-6 cycloalkyl, -S(＝O) ₂ -(C _1-6 alkyl), -S(＝O) ₂ -(C _3-6 cycloalkyl), -P(＝O)(C _1-6 alkyl) ₂ , (-C _3-6 cycloalkylene)-CN and (-C _3-6 cycloalkylene)-C _1-6 haloalkyl. The compound of any one of claims 1 to 9, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein R ² is selected from: A compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, wherein X is a benzene ring or a 5-6 membered heteroaromatic ring (e.g., a pyridine ring); and Y is absent; R ²³ is independently selected at each occurrence from H, halogen, -SF ₅ , C _1-6 alkyl, haloC _1-6 alkyl, C _3-6 cycloalkyl, -S(═O) ₂ -(C _1-6 alkyl), -S(═O) ₂ -(C _3-6 cycloalkyl), -P(═O)(C _1-6 alkyl) ₂ , (-C _3-6 cycloalkylene)-CN and (-C _3-6 cycloalkylene)-C _1-6 haloalkyl; Preferably, ^R2 is selected from: A compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, wherein R ³ is H or C _1-6 alkyl; preferably ethyl. The compound of any one of claims 1 to 12, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, wherein ring Z is a 3-10 membered heterocycle or a benzene ring; preferably a 5-10 membered heterocycle; more preferably a 5-6 membered heterocycle; and The heterocyclic ring and the benzene ring are each optionally substituted at each occurrence by one or more substituents independently selected from the group consisting of halogen and C _1-6 alkyl; Preferably, ring Z is A compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein L ² is -C(=O)- or -NRC(=O)-, wherein R is H or C _1-6 alkyl; Preferably, ^L2 is -C(=O)-. A compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, wherein R ⁴¹ is selected from a 3-10 membered heterocyclic ring, a C _6-10 aromatic ring and a 5-14 membered heteroaromatic ring, wherein the heterocyclic ring, aromatic ring and heteroaromatic ring are each optionally substituted with one or more substituents independently selected from the following: halogen, -OH, -CN, C _1-6 alkyl, -OC _1-6 alkyl and -SC _1-6 alkyl, preferably, the heterocyclic ring, aromatic ring and heteroaromatic ring are at least substituted with -OH, -CN or -OC _1-6 alkyl; Preferably, -L ² -R ⁴¹ is The compound of any one of claims 1 to 15, or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, wherein R ⁴¹ is a 5-6 membered heteroaromatic ring (preferably a pyrazole ring or a pyrimidine ring), which is substituted with at least one -OH; Preferably, R ⁴¹ is a 5-membered heteroaromatic ring (preferably a pyrazole ring), which is substituted by at least one -OH; Preferably, -L ² -R ⁴¹ is Preferably, -L ² -R ⁴¹ is A compound according to any one of claims 1 to 16 or a pharmaceutically acceptable salt, ester, stereoisomer, atropisomer or the like isomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug, wherein the compound is selected from:


















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

A pharmaceutical composition comprising a preventive or therapeutically effective amount of a compound according to any one of claims 1 to 18 or a pharmaceutically acceptable The invention relates to pharmaceutical compositions and methods comprising the use of pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, metabolites, isotope-labeled compounds or prodrugs, and pharmaceutically acceptable carriers. Use of a compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite, isotope-labeled compound or prodrug thereof, or a pharmaceutical composition according to claim 19 in the preparation of a medicament for use as a WRN inhibitor, preferably, the medicament is used to prevent or treat cancer (preferably, the cancer is characterized by microsatellite high instability (MSI-H) or mismatch repair deficiency (dMMR)); preferably, the cancer is selected from colorectal cancer, gastric cancer, endometrial cancer, uterine cancer, adrenocortical carcinoma, cervical cancer, esophageal cancer, breast cancer, kidney cancer, prostate cancer and ovarian cancer.