WO2025077867A1 - Pyridine polycyclic derivative and use thereof - Google Patents

Abstract

The present application relates to the field of medicines, and in particular to a pyridone polycyclic derivative and a use thereof, and particularly discloses a compound as shown in formula (I).

Description

Pyridone cyclic derivatives and their applications

The present invention claims the following priority

CN2023113213541, Application Date: October 12, 2023,

CN2024106577069, application date: May 26, 2024.

Technical Field

The present invention relates to the field of medicine, and in particular to pyridone cyclopentane derivatives and applications thereof.

Background Art

Helicases are a class of enzymes that can unwind double-stranded nucleotides. They were discovered in Escherichia coli in 1976. Helicases are widely present in a variety of organisms. Although there are many types, this type of protease has certain homology and similarity in its amino acid sequence. There are five types of helicases in the human body: RECQ1, BLM, WRN, RECQ4, and RECQ5. Among them, defects in the genes encoding BLM, WRN, and RECQ4 can lead to corresponding human diseases, namely Bloom syndrome (BS), Werner syndrome (WS), Rothmund-Thomson syndrome (RTS), RAPADIINO syndrome, and Baller-Gerold syndrome. These diseases are extremely rare recessive genetic diseases in humans, and at the molecular level, they are manifested as high genome instability, chromosomal abnormalities (chromosome breakage and deletion, rearrangement, sister chromosome exchange, etc.), and increased sensitivity to DNA damage factors. Clinically, they manifest as premature aging, type 2 diabetes, osteoporosis, arteriosclerosis, and extreme susceptibility to cancer.

WRN helicase is composed of 1432 amino acids and has a molecular mass of 160KD. In undamaged human cells, WRN is mainly localized in the nucleolus, but after DNA damage, it will quickly move to other nuclear regions. Immunochemical staining of S-phase cells showed that WRN was distributed in a dot-like pan-nuclear distribution, indicating that WRN may be related to DNA replication. WRN is the only member of the human RecQ helicase family with 3, → 5, exonuclease activity. Its N-terminal exonuclease region is conducive to WRN processing complex nucleic acid structures. The middle unwinding domain can bind and hydrolyze ATP to provide energy to unwind double-stranded DNA. There is an acidic region with transcriptional activity between the exonuclease and helicase regions. Immediately following the helicase region, there is a highly conserved RQC domain that plays a role in stabilizing the structure and recognizing and binding DNA. The C-terminal HRDC domain can regulate the activity of catalytic binding to DNA.

In addition to its helicase and exonuclease activities, WRN also interacts with many proteins in the BER pathway, including POLB, POLD, proliferating cell nuclear antigen (PCNA), replication protein A (RPA), structure-specific endonuclease 1 (FEN-1), and poly (ADP-ribose) polymerase 1 (PARP-1). WRN facilitates the synthesis of hairpin structures and trinucleotide repeat sequence quadruplex structures by POLD, which plays an important role in DNA damage repair.

Loss of DNA mismatch repair function is a key driver of cancer development and progression, occurring in 10-30% of colorectal, endometrial, ovarian, and gastric cancers. Cancers that lose mismatch repair (MMR) have a high mutational burden, with new microsatellite alleles appearing at a microsatellite locus in tumors due to insertion or deletion of repeat units compared to normal tissues. This change in the size of microsatellite loci has been shown to be caused by defective DNA mismatch repair (dMMR), a phenomenon known as microsatellite instability (MSI). Although immunotherapy has shown significant efficacy in microsatellite unstable colorectal cancer, approximately 60% of MSI colorectal cancer patients do not benefit from it. In tumor cell lines with high microsatellite instability (MSI), knockout of the WRN gene or depletion of the WRN protein results in synthetic lethality. The synthetic lethal interaction of WRN is specifically associated with MMR (DNA mismatch repair) defects. Due to the defective MMR system, these cancer cells exhibit high MSI, i.e., frequent mutations in microsatellite repeat sequences in DNA. Loss of WRN leads to genomic instability, cell cycle arrest, DNA double-strand breaks, etc. in these cancer cells, ultimately leading to cancer cell death. Mechanistically, a protein complex of the structure-specific endonuclease subcomplex MUS81-EME1 and the scaffold protein SLX4 is the main cause of double-strand DNA breaks around (TA) repeat sequences in high-MSI cells. The generation of non-B-form DNA secondary structures in genomic regions induces the detection of stalled replication forks around (TA) repeat loci, which triggers the activation of the phosphorylation-based checkpoint kinase ATR, which subsequently recruits WRN to resolve stalled forks through its helicase activity. Notably, (TA) repeat regions are repeat-expanded in high-MSI cells, leading to the accumulation of non-B-form DNA structures throughout the genome, explaining the importance of WRN in these cells.

Preclinical studies have confirmed that WRN is a synthetic lethal target in dMMR/MSI-H colorectal cancer tumors and can be used as a potential treatment method alone or in combination with targeted drugs, chemotherapy or immunotherapy. Therefore, WRN inhibitors can achieve selective killing of MSI cancer cells, providing the possibility for precise tumor treatment. Currently, only Novartis' WRN small molecule inhibitor HRO761 has entered Phase 1 clinical trials. There is a huge market prospect for developing new WRN inhibitors with better efficacy and higher safety for the treatment of tumors.

The present invention provides pyridone cyclopentadienyl compounds, and their analogs and derivatives, for inhibiting the activity of WRN protein and methods for treating diseases using the compounds, in particular for treating cancers with high microsatellite instability (MSI-H) or mismatch repair deficiency (dMMR), including colorectal cancer, gastric cancer, endometrial cancer, lung cancer, head and neck cancer, liver cancer, breast cancer, melanoma, renal cell carcinoma, etc.

Summary of the invention

The first aspect of the present application provides a compound of formula (I) or a pharmaceutically acceptable salt thereof,

in,

Ring A is selected from 5-6 membered heteroaryl, benzene ring and 5-6 membered heterocycloalkenyl;

L ₁ is selected from

Each R ₁ is independently selected from H, C _1-3 alkyl, C _2-3 alkenyl, C _2-3 alkynyl, or two R ₁ on adjacent carbon atoms together with the carbon atom to which they are connected form a 3-10 membered ring, or two R ₁ on two non-adjacent carbon atoms form a bridge containing 1 or 2 carbon atoms, the C _1-3 alkyl group is optionally substituted by 1, 2, 3 or 4 R _a , the C _2-3 alkenyl, C _2-3 alkynyl, and the 3-10 membered ring is optionally substituted with 1, 2, 3 or 4 R _b ;

each R ₂ is independently selected from H, F, Cl, Br, NH ₂ , CN, OH and C _1-3 alkyl, wherein the C _1-3 alkyl is optionally substituted with 1, 2, 3 or 4 R _c ;

_R3 is selected from _C1-3 alkyl;

R ₄ is selected from

R _a is selected from

R _b is selected from H, F, Cl, Br, NH ₂ , CN, OH and C _1-3 alkyl;

R _c is selected from H, F, Cl, Br, NH ₂ , CN, OH and C _1-3 alkyl;

n is selected from 0, 1, 2, 3 and 4;

m is selected from 0, 1, 2, 3 and 4;

The condition is,

When ring A is selected from 5-6 membered heterocycloalkenyl, _L1 is selected from When R ₄ is not

In the preferred embodiment of the present application, wherein _Ra is selected from Other variables are as defined in this application.

In the preferred embodiment of the present application, wherein _Ra is selected from Other variables are as defined in this application.

In a preferred embodiment of the present application, R _b is selected from H, F, Cl, Br, NH ₂ , CN, OH, CH ₃ , CH ₂ CH ₃ and CH ₂ CH ₂ CH ₃ , and other variables are as defined in the present application.

In a preferred embodiment of the present application, R _c is selected from H, F, Cl, Br, NH ₂ , CN, OH, CH ₃ , CH ₂ CH ₃ and CH ₂ CH ₂ CH ₃ , and other variables are as defined in the present application.

In the preferred embodiment of the present application, wherein L ₁ is selected from Other variables are as defined in this application.

In the preferred embodiment of the present application, wherein L ₁ is selected from Other variables are as defined in this application.

In the preferred embodiment of the present application, wherein L ₁ is selected from Other variables are as defined in this application.

In a preferred embodiment of the present application, wherein ring A is selected from Other variables are as defined in this application.

In a preferred embodiment of the present application, each R ₁ is independently selected from H, CH ₃ , CH ₂ CH ₃ , CH ₂ CH ₂ CH ₃ , CH _{2 ═CH 2} , HC═CH or two R ₁ on two adjacent carbon atoms together with the carbon atom to which they are attached form a 3-membered ring, a 4-membered ring, a 5-membered ring, a 6-membered ring, a 7-membered ring, an 8-membered ring, a 9-membered ring and a 10-membered ring, the CH ₃ , CH ₂ CH ₃ and CH _{2 CH 2 CH 3} are optionally substituted with 1, 2, 3 or 4 R _a , the CH ₂ ＝CH ₂ , HC≡CH, a 3-membered ring, a 4-membered ring, a 5-membered ring, a 6-membered ring, a 7-membered ring, an 8-membered ring, a 9-membered ring and a 10-membered ring are optionally substituted with 1, 2, 3 or 4 R _b , and other variables are as defined in the present application.

In the preferred embodiment of the present application, wherein the above-mentioned 3-membered ring is selected from Other variables are as defined in this application.

In the preferred embodiment of the present application, the 4-membered ring is selected from Other variables are as defined in this application.

In the preferred embodiment of the present application, the 5-membered ring is selected from Other variables are as defined in this application.

In the preferred embodiment of the present application, the 6-membered ring is selected from Other variables are as defined in this application.

In a preferred embodiment of the present application, the 7-membered ring is selected from a monocyclic ring, a spirocyclic ring, a fused ring and a bridged ring, and other variables are as defined in the application.

In the preferred embodiment of the present application, the 7-membered ring is selected from Other variables are as defined in the application.

In a preferred embodiment of the present application, the 8-membered ring is selected from a monocyclic ring, a spirocyclic ring, a fused ring and a bridged ring, and other variables are as defined in the application.

In a preferred embodiment of the present application, the 9-membered ring is selected from a monocyclic ring, a spirocyclic ring, a fused ring and a bridged ring, and other variables are as defined in the application.

In the preferred embodiment of the present application, wherein the above 9-membered ring is selected from Other variables are as defined in the application.

In a preferred embodiment of the present application, the 10-membered ring is selected from a monocyclic ring, a spirocyclic ring, a fused ring and a bridged ring, and other variables are as defined in the application.

In a preferred embodiment of the present application, each R ₁ is independently selected from H, CH ₃ , CH ₂ CH ₃ , CH ₂ CH ₂ CH ₃ , CH _{2 ═CH 2} , HC═CH Or two R ₁ on two adjacent carbon atoms together with the carbon atom to which they are attached form The CH ₃ , CH ₂ CH ₃ and CH ₂ CH ₂ CH ₃ are optionally substituted by 1, 2, 3 or 4 _Ra , and the CH ₂ ＝CH ₂ , HC≡CH, Optionally substituted with 1, 2, 3 or 4 R _b , and the other variables are as defined herein.

In the preferred embodiment of the present application, the structural unit Selected from Other variables are as defined in this application.

In a preferred embodiment of the present application, R ₃ is selected from CH ₃ , CH ₂ CH ₃ and CH ₂ CH ₂ CH ₃ , and other variables are as defined in the present application.

In the preferred embodiment of the present application, wherein R ₄ is selected from Other variables are as defined in this application.

In the preferred embodiment of the present application, wherein R ₄ is selected from Other variables are as defined in this application.

In the preferred embodiment of the present application, wherein R ₄ is selected from Other variables are as defined in this application.

In the preferred embodiment of the present application, wherein R ₄ is selected from Other variables are as defined in this application.

In a preferred embodiment of the present application, the compound is selected from

wherein Ring A, R ₁ , R ₂ , R ₃ , R ₄ , L ₁ , n and m are as defined herein.

Some technical solutions of the present application are obtained by freely combining the above variables.

The present application provides the following compounds or pharmaceutically acceptable salts thereof,

The second invention aspect of the present application provides use of the compound defined in any of the above technical solutions or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating diseases associated with WRN.

In a preferred embodiment of the present application, the WRN-related disease refers to cancer with high microsatellite instability (MSI-H) or mismatch repair deficiency (dMMR), and is further preferably colorectal cancer, gastric cancer, endometrial cancer, lung cancer, head and neck cancer, liver cancer, breast cancer, melanoma, and renal cell carcinoma.

Definition and Explanation

Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered to be uncertain or unclear in the absence of a special definition, but should be understood according to its ordinary meaning. When a trade name appears in this article, it is intended to refer to its corresponding commercial product or its active ingredient.

The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and/or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit/risk ratio.

The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present application, prepared from a compound with a specific substituent found in the present application and a relatively non-toxic acid or base. When a relatively acidic functional group is contained in the compound of the present application, a base addition salt can be obtained by contacting such a compound with a sufficient amount of a base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts. When a relatively basic functional group is contained in the compound of the present application, an acid addition salt can be obtained by contacting such a compound with a sufficient amount of an acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts, and also include salts of amino acids (such as arginine, etc.), and salts of organic acids such as glucuronic acid. Certain specific compounds of the present application contain basic and acidic functional groups, and can thus be converted into any base or acid addition salt.

The pharmaceutically acceptable salts of the present application can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. Generally, the preparation method of such salts is: in water or an organic solvent or a mixture of the two, these compounds in free acid or base form are reacted with a stoichiometric amount of an appropriate base or acid to prepare.

The compounds of the present application may exist in specific geometric or stereoisomeric forms. The present application contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as mixtures enriched in enantiomers or diastereomers, all of which fall within the scope of the present application. Additional asymmetric carbon atoms may exist in substituents such as alkyl. All of these isomers and their mixtures are included within the scope of the present application.

Unless otherwise indicated, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of one another.

Unless otherwise indicated, the term "cis-trans isomers" or "geometric isomers" arises from the inability of a ring to rotate freely about double bonds or single bonds of ring carbon atoms.

Unless otherwise indicated, the term "diastereomer" refers to stereoisomers that have two or more chiral centers and that are not mirror images of each other.

Unless otherwise indicated, "(+)" indicates dextrorotatory, "(-)" indicates levorotatory, and "(±)" indicates racemic.

Unless otherwise specified, the key is a solid wedge. and dotted wedge key To indicate the absolute configuration of a stereocenter, use a straight solid bond. and straight dashed key To indicate the relative configuration of a stereocenter, use a wavy line Indicates a wedge-shaped solid key or dotted wedge key Or use a wavy line Represents a straight solid bond and straight dashed key

Unless otherwise indicated, the term "tautomer" or "tautomeric form" refers to isomers of different functional groups that are in dynamic equilibrium at room temperature and can readily interconvert. If tautomerism is possible (such as in solution), chemical equilibrium of the tautomers can be achieved. For example, proton tautomers (also called prototropic tautomers) include interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions via reorganization of some of the bonding electrons. A specific example of keto-enol tautomerism is the interconversion between pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

Unless otherwise indicated, the terms "enriched in one isomer", "isomerically enriched", "enriched in one enantiomer" or "enantiomerically enriched" mean that the content of one isomer or enantiomer is less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

Unless otherwise indicated, the term "isomer excess" or "enantiomeric excess" refers to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, the isomer or enantiomeric excess (ee value) is 80%.

Optically active (R)- and (S)-isomers and D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present application is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are separated by conventional methods known in the art, and then the pure enantiomer is recovered. In addition, the separation of enantiomers and diastereomers is usually accomplished by using chromatography, which uses a chiral stationary phase and is optionally combined with a chemical derivatization method (e.g., carbamate is generated from an amine).

The compounds of the present application may contain non-natural proportions of atomic isotopes on one or more atoms constituting the compound. For example, compounds may be labeled with radioactive isotopes, such as tritium ( ^3H ), iodine-125 ( ^125I ) or C-14 ( ^14C ). For another example, deuterated drugs may be formed by replacing hydrogen with heavy hydrogen. The bond formed by deuterium and carbon is stronger than the bond formed by ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs have the advantages of reducing toxic side effects, increasing drug stability, enhancing therapeutic effects, and extending the biological half-life of drugs. All isotopic composition changes of the compounds of the present application, whether radioactive or not, are included in the scope of the present application.

"Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include deuterium and hydrogen variants, as long as the valence state of the particular atom is normal and the substituted compound is stable. The type and number of substituents can be any on the basis of chemical achievable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups.

When any variable (e.g., R ₁ ) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 0-2 R _{1 s} , the group may be optionally substituted with up to two R _{1 s} , and each occurrence of R ₁ is an independent choice. In addition, combinations of substituents and/or variants thereof are permitted only if such combinations result in stable compounds.

When the number of a linking group is 0, such as -(CRR) ₀ -, it means that the linking group is a single bond.

When one of the variables is selected from a single bond, it means that the two groups it connects are directly connected. For example, when L in A-L-Z represents a single bond, it means that the structure is actually A-Z.

When a substituent is vacant, it means that the substituent does not exist. For example, when X in A-X is vacant, it means that the structure is actually A. When the listed substituent does not specify which atom it is connected to the substituted group through, the substituent can be bonded through any atom of it. For example, pyridyl as a substituent can be connected to the substituted group through any carbon atom on the pyridine ring.

When the listed linking group does not indicate its linking direction, its linking direction is arbitrary, for example, Medium connection The group L is -MW-, in which case -MW- can be connected to ring A and ring B in the same direction as the reading order from left to right to form You can also connect ring A and ring B in the opposite direction of the reading order from left to right to form Combinations of linkers, substituents, and/or variations thereof are permissible only if such combinations result in stable compounds.

Unless otherwise specified, when a group has one or more connectable sites, any one or more sites of the group can be connected to other groups through chemical bonds. When the chemical bond connection mode is non-positional and there are H atoms at the connectable sites, when the chemical bonds are connected, the number of H atoms at the site will decrease with the number of connected chemical bonds to become a group with the corresponding valence. The chemical bond connecting the site to other groups can be a straight solid bond. Straight dotted key or wavy line For example, the straight solid bond in -OCH ₃ indicates that it is connected to other groups through the oxygen atom in the group; The straight dashed bond in the group indicates that the two ends of the nitrogen atom in the group are connected to other groups; The wavy line in the phenyl group indicates that it is connected to other groups through the carbon atoms at positions 1 and 2 in the phenyl group; It means that any connectable site on the piperidine group can be connected to other groups through one chemical bond, including at least These four connection methods, even if the H atom is drawn on -N-, Still includes For groups connected in this way, when one chemical bond is connected, the H at that site will be reduced by one and become a corresponding monovalent piperidine group.

When a non-positioning substituent is attached to a polycyclic system, as follows In the structure shown in FIG. 1 , a substituent group connected to only one ring is connected to only the ring. For example, in the structure shown in FIG. 1 , a substituent group connected to only one ring is connected to only the ring. It is only connected to ring _B1 but not to ring _B2 ; when the connecting bond penetrates all rings in the ring system, it means that the substituent can be substituted to all rings in the ring system. For example, in the above structure, _R2 can be substituted to both ring _B1 and ring _B2 .

Unless otherwise specified, the number of atoms in a ring is generally defined as the ring member number, for example, "3-7 membered ring" refers to a "ring" having 3-7 atoms arranged around it.

Alicyclic refers to a saturated or partially unsaturated all-carbon ring system. Wherein "partially unsaturated" refers to a ring portion including at least one double bond or triple bond, and "partially unsaturated" is intended to cover rings with multiple unsaturated sites, but is not intended to include aryl or heteroaryl moieties as defined herein. Non-limiting examples include cyclopropyl rings, cyclobutyl rings, cyclopentyl rings, cyclopentenyl rings, cyclohexyl rings, cyclohexenyl rings, cyclohexadienyl rings, cycloheptyl rings, cycloheptatrienyl rings, cyclopentanone rings, cyclopentane-1,3-dione rings, etc.

Alicyclic group refers to a saturated or partially unsaturated alicyclic group in which 1, 2 or 3 ring carbon atoms are replaced by heteroatoms selected from nitrogen, oxygen or S(O) _t (wherein t is an integer from 0 to 2), but does not include the ring portion of -OO-, -OS- or -SS-, and the remaining ring atoms are carbon. Non-limiting examples include an oxetane ring, an azetidine ring, an oxetane ring, a tetrahydrofuran ring, a tetrahydrothiophene ring, a tetrahydropyrrole ring, a piperidine ring, a pyrroline ring, an oxazolidine ring, a piperazine ring, a dioxolane ring, a dioxane ring, a morpholine ring, a thiomorpholine ring, a thiomorpholine-1,1-dioxide, a tetrahydropyran ring, an azetidine-2-one ring, an oxetane-2-one ring, a pyrrolidine-2-one ring, a pyrrolidine-2,5-dione ring, a piperidine-2-one ring, a dihydrofuran-2(3H)-one ring, a dihydrofuran-2,5-dione ring, a tetrahydro-2H-pyran-2-one ring, a piperazine-2-one ring, and a morpholine-3-one ring. Non-limiting examples of partially unsaturated monocyclic heterocycles include 1,2-dihydroazetidine ring, 1,2-dihydrooxetadiene ring, 2,5-dihydro-1H-pyrrole ring, 2,5-dihydrofuran ring, 2,3-dihydrofuran ring, 2,3-dihydro-1H-pyrrole ring, 3,4-dihydro-2H-pyran ring, 1,2,3,4-tetrahydropyridine ring, 3,6-dihydro-2H-pyran ring, 1,2,3,6-tetrahydropyridine ring, 4,5-dihydro-1H-imidazole ring, 1,4,5,6-tetrahydropyrimidine ring, 3,4,7,8-tetrahydro-2H-1,4,6-oxadiazolidine ring, 1,6-dihydropyrimidine ring, 4,5,6,7-tetrahydro-1H-1,3-diazepine Cyclic, 2,5,6,7-tetrahydro-1,3,5-oxadiazepine Ring, etc.

"Aryl" and "aromatic ring" are used interchangeably, and both refer to an all-carbon monocyclic or fused polycyclic (i.e., rings that share adjacent pairs of carbon atoms) group with a conjugated π electron system, which group may be fused with a cycloalkyl ring, a heterocycloalkyl ring, a cycloalkenyl ring, a heterocycloalkenyl ring, or a heteroaryl group. " _C6-10 aryl" refers to a monocyclic or bicyclic aromatic group having 6 to 10 carbon atoms, and non-limiting examples of aryl include phenyl, naphthyl, and the like.

"Heteroaryl" and "heteroaryl ring" are used interchangeably and refer to a group of a monocyclic, bicyclic or polycyclic 4n+2 aromatic ring system (e.g., having 6 or 10 π electrons shared in a cyclic arrangement) having ring carbon atoms and ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur. In the present application, heteroaryl also includes a ring system in which the above-mentioned heteroaryl ring is fused with one or more cycloalkyl rings, heterocycloalkyl rings, cycloalkenyl rings, heterocycloalkenyl rings or aromatic rings. The heteroaryl ring may be optionally substituted. "5 to 10 membered heteroaryl" refers to a monocyclic or bicyclic heteroaryl having 5 to 10 ring atoms, wherein 1, 2, 3 or 4 ring atoms are heteroatoms. "5- to 6-membered heteroaryl" refers to a monocyclic heteroaryl group having 5 to 6 ring atoms, wherein 1, 2, 3 or 4 of the ring atoms are heteroatoms, non-limiting examples of which include thienyl, furanyl, thiazolyl, isothiazolyl, imidazolyl, oxazolyl, pyrrolyl, pyrazolyl, triazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, tetrazolyl, isoxazolyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, tetrazinyl. "8- to 10-membered heteroaryl" refers to a bicyclic heteroaryl group having 8 to 10 ring atoms, wherein 1, 2, 3 or 4 of the ring atoms are heteroatoms, non-limiting examples of which include indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzisothiazolyl, The term "heteroatom" refers to nitrogen, oxygen or sulfur. In heteroaryl groups containing one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom, as valence permits. Heteroaryl bicyclic ring systems may include one or more heteroatoms in one or both rings.

Unless otherwise specified, the term "C _1-3 alkyl" is used to refer to a straight or branched saturated hydrocarbon group consisting of 1 to 3 carbon atoms, which may be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). Examples of C _1-3 alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl).

[0043] The terms "halo" or "halogen," by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom.

Unless otherwise specified, the terms "C _6-10 aryl" and "C _6-10 aromatic ring" can be used interchangeably. The term "C _6-10 aromatic ring" or "C _6-10 aryl" means a cyclic hydrocarbon group composed of 6 to 10 carbon atoms with a conjugated π electron system, which can be a monocyclic, fused bicyclic or fused tricyclic system, wherein each ring is aromatic. It can be monovalent, divalent or polyvalent, and C6-10 aryl includes C6-9, C9, C10 and C6 aryl, etc. Examples of C6-10 aryl include, but are not limited to, phenyl, naphthyl (including 1-naphthyl and 2-naphthyl, etc.).

Unless otherwise specified, the terms "5-10 membered heteroaromatic ring" and "5-10 membered heteroaryl" can be used interchangeably, and the term "5-10 membered heteroaryl" refers to a cyclic group consisting of 5 to 10 ring atoms with a conjugated π electron system, wherein 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S and N, and the rest are carbon atoms. It can be a monocyclic, fused bicyclic or fused tricyclic system, wherein each ring is aromatic. Wherein the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms can be optionally oxidized (i.e., NO and S(O)p, p is 1 or 2). The 5-10 membered heteroaryl can be connected to the rest of the molecule via a heteroatom or a carbon atom. The 5-10 membered heteroaryl includes 10-membered, 9-membered, 9-10-membered, 5-8-membered, 5-7-membered, 5-6-membered, 5-membered and 6-membered heteroaryl, etc. Examples of the 5-10 membered heteroaryl group include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl, etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl and 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl and 5-oxazolyl, etc.), triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl and 4H-1,2,4-triazolyl, etc.), tetrazolyl, isoxazolyl (3-isoxazolyl, 4-isoxazolyl and 5-isoxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4- thiazolyl and 5-thiazolyl, etc.), furanyl (including 2-furanyl and 3-furanyl, etc.), thienyl (including 2-thienyl and 3-thienyl, etc.), pyridyl (including 2-pyridyl, 3-pyridyl and 4-pyridyl, etc.), pyrazinyl, pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl, etc.), benzothiazolyl (including 5-benzothiazolyl, etc.), purinyl, benzimidazolyl (including 2-benzimidazolyl, etc.), benzoxazolyl, indolyl (including 5-indolyl, etc.), isoquinolyl (including 1-isoquinolyl and 5-isoquinolyl, etc.), quinoxalinyl (including 2-quinoxalinyl and 5-quinoxalinyl, etc.), or quinolyl (including 3-quinolyl and 6-quinolyl, etc.).

Unless otherwise specified, the terms "5-6 membered heteroaromatic ring" and "5-6 membered heteroaryl" can be used interchangeably, and the term "5-6 membered heteroaryl" means a monocyclic group consisting of 5 to 6 ring atoms with a conjugated π electron system, 1, 2, 3 or 4 of which are heteroatoms independently selected from O, S and N, and the rest are carbon atoms. Wherein the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms can be optionally oxidized (i.e., NO and S(O)p, p is 1 or 2). The 5-6 membered heteroaryl can be connected to the rest of the molecule through a heteroatom or a carbon atom. The 5-6 membered heteroaryl includes 5-membered and 6-membered heteroaryl. Examples of the 5-6 membered heteroaryl group include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl, etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl and 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl and 5-oxazolyl, etc.), triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl) and 4H-1,2,4-triazolyl, etc.), tetrazolyl, isoxazolyl (3-isoxazolyl, 4-isoxazolyl and 5-isoxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4-thiazolyl and 5-thiazolyl, etc.), furanyl (including 2-furanyl and 3-furanyl, etc.), thienyl (including 2-thienyl and 3-thienyl, etc.), pyridyl (including 2-pyridyl, 3-pyridyl and 4-pyridyl, etc.), pyrazinyl or pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl, etc.).

Unless otherwise specified, the term "3-10 membered ring" refers to a saturated, unsaturated or partially unsaturated cyclic group consisting of 3 to 10 ring atoms, including monocyclic and bicyclic systems, wherein the bicyclic system includes spirocyclic, fused and bridged rings. The "3-10 membered ring" is selected from C _3-10 cycloalkyl, 3-10 membered heterocycloalkyl, C _3-10 cycloalkenyl, 3-10 membered heterocycloalkenyl, C _6-10 aryl and 5-10 membered heteroaryl, and the C _3-10 cycloalkyl, 3-10 membered heterocycloalkyl, C _3-10 cycloalkenyl, 3-10 membered heterocycloalkenyl, C _6-10 aryl and 5-10 membered heteroaryl are as defined in the present application.

Unless otherwise specified, the term "C _3-10 cycloalkyl" means a saturated cyclic hydrocarbon group consisting of 3 to 10 carbon atoms, including monocyclic and bicyclic systems, wherein the bicyclic system includes spirocyclic, fused and bridged rings. The C _3-7 cycloalkyl includes C _3-6 , C _4-6 , C _4-5 , C _5-7 or C _5-6 cycloalkyl, etc.; it can be monovalent, divalent or polyvalent. Examples of C _3-10 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, etc.

Unless otherwise specified, the term "C _3-10 monocycloalkyl" means a saturated single cyclic hydrocarbon group consisting of 3 to ₁₀ carbon atoms, and the C _3-10 monocycloalkyl includes C _3-6 , C _4-6 , C _4-5 , C _5-7 or C _5-6 monocycloalkyl, etc.; it can be monovalent, divalent or polyvalent. Examples of C _3-10 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, etc.

Unless otherwise specified, the term "3-10 membered heterocycloalkyl" by itself or in combination with other terms refers to a saturated cyclic group consisting of 3 to 10 ring atoms, 1, 2, 3 or 4 of which are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms may be optionally oxidized (i.e., NO and S(O) _p , p is 1 or 2). It includes monocyclic and bicyclic ring systems, wherein the bicyclic ring system includes spirocyclic, paracyclic and bridged rings. In addition, with respect to the "3-10 membered heterocycloalkyl", heteroatoms may occupy the position where the heterocycloalkyl is connected to the rest of the molecule. The 3-10 membered heterocycloalkyl includes 3-4 membered, 4-5 membered, 5-7 membered, 3 membered, 4 membered, 5 membered, 6 membered, 7 membered, 8-9 membered, 8-10 membered heterocycloalkyl, etc. Examples of 3-10 membered heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl or hexahydropyridazinyl, etc.

Unless otherwise specified, the term "3-10 membered monoheterocycloalkyl" by itself or in combination with other terms means a single saturated cyclic group consisting of 3 to 10 ring atoms, which is a monocyclic ring system, and the other definitions are the same as 3-10 membered heterocycloalkyl.

Unless otherwise specified, the term "C _3-10 cycloalkenyl" by itself or in combination with other terms means a partially unsaturated cyclic hydrocarbon group consisting of 3 to 10 ring atoms containing at least one carbon-carbon double bond, including monocyclic and bicyclic systems, wherein the bicyclic system includes spiro, fused and bridged rings. The C _3-10 cycloalkenyl includes C _3-6 , C _4-6 , C _4-5 , C _5-7 or C _5-6 cycloalkenyl, etc.; it can be monovalent, divalent or polyvalent. Examples of C _3-10 cycloalkyl include, but are not limited to

Unless otherwise specified, the term "3-10 membered heterocycloalkenyl" by itself or in combination with other terms means a partially unsaturated cyclic group consisting of 3 to 10 ring atoms containing at least one carbon-carbon double bond, wherein 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms may be optionally oxidized (i.e., NO and S(O)p, p is 1 or 2). It includes monocyclic and bicyclic ring systems, wherein the bicyclic ring system includes spirocyclic, paracyclic and bridged rings, and any ring of this system is non-aromatic. In addition, with respect to the "3-10 membered heterocycloalkenyl", heteroatoms may occupy the connection position of the heterocycloalkenyl group to the rest of the molecule. The 3-10 membered heterocycloalkenyl group includes 3-membered, 4-membered, 5-membered and 6-membered heterocycloalkenyl groups, etc. Examples of 3-10 membered heterocycloalkenyl groups include, but are not limited to

Unless otherwise specified, the term "5-6 membered heterocycloalkenyl" by itself or in combination with other terms means a partially unsaturated cyclic group consisting of 5 to 6 ring atoms containing at least one carbon-carbon double bond, wherein 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms may be optionally oxidized (i.e., NO and S(O)p, p is 1 or 2). It includes monocyclic and bicyclic ring systems, wherein the bicyclic ring system includes spirocyclic, paracyclic and bridged rings, and any ring of this system is non-aromatic. In addition, with respect to the "5-6 membered heterocycloalkenyl", heteroatoms may occupy the position where the heterocycloalkenyl is connected to the rest of the molecule. Examples of the 5-6 membered heterocycloalkenyl include, but are not limited to

Unless otherwise specified, the terms "5-membered heteroaromatic ring" and "5-membered heteroaryl" are used interchangeably in this application. The term "5-membered heteroaryl" refers to a monocyclic group with a conjugated π electron system consisting of 5 ring atoms, 1, 2, 3 or 4 of which are heteroatoms independently selected from O, S and N, and the rest are carbon atoms. The nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O) _p , p is 1 or 2). The 5-membered heteroaryl can be connected to the rest of the molecule through a heteroatom or a carbon atom. Examples of the 5-membered heteroaryl group include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl, etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl and 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl and 5-oxazolyl, etc.), triazolyl (1H-1,2,3-triazolyl, 2 H-1,2,3-triazolyl, 1H-1,2,4-triazolyl and 4H-1,2,4-triazolyl, etc.), tetrazolyl, isoxazolyl (3-isoxazolyl, 4-isoxazolyl and 5-isoxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4-thiazolyl and 5-thiazolyl, etc.), furanyl (including 2-furanyl and 3-furanyl, etc.), thienyl (including 2-thienyl and 3-thienyl, etc.).

Unless otherwise specified, Cn _-n+m or _Cn - _Cn+m includes any specific case of n to n+m carbon atoms, for example, _C1-12 includes _C1 , _C2 , C3, _C4 , _{C5 , C6, C7, C8, C9, C10, C11 , and C12 , and also includes} any range from n to n+m, for example _{, C1-12} includes _C1-3 , _C1-6 , _C1-9 , _C3-6 , _C3-9 , _C3-12 , _C6-9 , _C6-12 , and C13. _9-12 , etc.; similarly, n-membered to n+m-membered means that the number of atoms in the ring is n to n+m, for example, 3-12-membered ring includes 3-membered ring, 4-membered ring, 5-membered ring, 6-membered ring, 7-membered ring, 8-membered ring, 9-membered ring, 10-membered ring, 11-membered ring, and 12-membered ring, and also includes any range from n to n+m, for example, 3-12-membered ring includes 3-6-membered ring, 3-9-membered ring, 5-6-membered ring, 5-7-membered ring, 6-7-membered ring, 6-8-membered ring, and 6-10-membered ring, etc.

The term "leaving group" refers to a functional group or atom that can be replaced by another functional group or atom through a substitution reaction (e.g., a nucleophilic substitution reaction). For example, representative leaving groups include trifluoromethanesulfonate; chlorine, bromine, iodine; sulfonate groups, such as mesylate, tosylate, p-brosylate, p-toluenesulfonate, etc.; acyloxy groups, such as acetoxy, trifluoroacetoxy, etc.

The term "protecting group" includes, but is not limited to, "amino protecting group", "hydroxy protecting group" or "thiol protecting group". The term "amino protecting group" refers to a protecting group suitable for preventing side reactions at the amino nitrogen position. Representative amino protecting groups include, but are not limited to: formyl; acyl, such as alkanoyl (such as acetyl, trichloroacetyl or trifluoroacetyl); alkoxycarbonyl, such as tert-butyloxycarbonyl (Boc); arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl, such as benzyl (Bn), trityl (Tr), 1,1-bis-(4'-methoxyphenyl)methyl; silyl, such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), etc. The term "hydroxy protecting group" refers to a protecting group suitable for preventing side reactions of the hydroxyl group. Representative hydroxy protecting groups include, but are not limited to, alkyl groups such as methyl, ethyl and tert-butyl; acyl groups such as alkanoyl (e.g., acetyl); arylmethyl groups such as benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (diphenylmethyl, DPM); silyl groups such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), and the like.

Unless otherwise defined, the "substituents independently selected from..." mentioned herein means that when more than one hydrogen on a group is replaced by a substituent, the substituents may be the same or different in type, and the substituents selected are independently of each other.

Generally, the compounds of the present application or their pharmaceutically acceptable salts, or their stereoisomers can be used in a suitable dosage form with one or more pharmaceutical carriers. These dosage forms are suitable for oral, rectal, topical, oral and other parenteral administration (e.g., subcutaneous, intramuscular, intravenous, etc.). For example, dosage forms suitable for oral administration include capsules, tablets, granules and syrups. The compounds of the present application contained in these preparations can be solid powders or particles; solutions or suspensions in aqueous or non-aqueous liquids; water-in-oil or water-in-oil emulsions, etc. The above dosage forms can be made from active compounds and one or more carriers or excipients via a common pharmaceutical method. The above carriers need to be compatible with the active compounds or other excipients. For solid preparations, commonly used non-toxic carriers include but are not limited to mannitol, lactose, starch, magnesium stearate, cellulose, glucose, sucrose, etc. Carriers for liquid preparations include water, saline, aqueous glucose solution, ethylene glycol and polyethylene glycol, etc. The active compound can form a solution or suspension with the above carriers.

The compositions of the present application are formulated, dosed and administered in a manner consistent with medical practice. The "therapeutically effective amount" of the compound administered is determined by factors such as the specific condition to be treated, the individual being treated, the cause of the condition, the target of the drug, and the mode of administration.

"Therapeutically effective amount" refers to the amount of the compound of the present invention that will induce a biological or medical response in an individual, such as reducing or inhibiting enzyme or protein activity or improving symptoms, alleviating symptoms, slowing or delaying disease progression, or preventing disease.

The therapeutically effective amount of the compound of the present application or its pharmaceutically acceptable salt, or its stereoisomer contained in the pharmaceutical composition or the pharmaceutical composition of the present application is preferably 0.1 mg-5 g/kg (body weight).

"Patient" refers to an animal, preferably a mammal, more preferably a human. The term "mammal" refers to warm-blooded vertebrate mammals, including, for example, cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, mice, pigs and humans.

"Treatment" means to lessen, slow the progression, attenuate, prevent, or maintain an existing disease or condition (eg, cancer). Treatment also includes curing, preventing the development of, or alleviating to some extent, one or more symptoms of a disease or condition.

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

The structure of the compounds of the present application can be confirmed by conventional methods known to those skilled in the art. If the present application involves the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the art. For example, single crystal X-ray diffraction (SXRD) is used to collect diffraction intensity data of the cultured single crystal using a Bruker D8 venture diffractometer, with the light source being CuKα radiation and the scanning mode being: After scanning and collecting relevant data, the crystal structure is further analyzed using the direct method (Shelxs97) to confirm the absolute configuration.

The solvents used in the present application can be obtained commercially. The following abbreviations are used in the present application: FA represents formic acid; NH ₂ CN represents cyanamide; K ₂ CO ₃ represents potassium carbonate; DMF represents N,N-dimethylformamide; prep-HPLC represents preparative high performance liquid chromatography; HCl represents hydrochloric acid; PE represents petroleum ether; EA represents ethyl acetate; Lawessons reagent represents Lawesson's reagent; DPPP represents bis(diphenylphosphino)propane; DBU represents 1,8-diazabicyclo[5.4.0]undec-7-ene; psi is a unit of pressure, representing pounds-force per square inch.

Compounds are named according to the conventional nomenclature in the art or using The software names were used, and commercially available compounds were named using the supplier's catalog names.

Beneficial Effects

The compounds of the present application have WRN protein inhibitory activity, and their SW48 cell anti-proliferation activity IC ₅₀ (nM) is 1-200 nM, preferably 1-100 nM, further preferably 5-50 nM, more preferably 5-30 nM, and most preferably 10-25 nM.

DETAILED DESCRIPTION

The present application is described in detail below through examples, but it is not intended to limit the present application in any adverse way. The present application has been described in detail herein, and its specific embodiments are also disclosed. It will be obvious to those skilled in the art that various changes and improvements can be made to the specific embodiments of the present application without departing from the spirit and scope of the present application.

Preparation Example 1: Synthesis of Intermediate INT D

Synthesis of Compound 1-2

At room temperature, compound 1-1 (30 g, 167 mmol) was dissolved in dichloroethane (300 mL), aluminum chloride (33.5 g, 251 mmol, 13.7 mL) was added at 0°C, and stirred at 40°C for 5 hours. Water (200 mL) was added to the system, and extracted with ethyl acetate (80 mL*3) for 3 times. The organic phases were combined, washed with saturated sodium chloride aqueous solution (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain crude compound 1-2, which was directly used in the next step.

LCMS:MS(ESI)m/z＝164.9[M+H] ⁺

¹ H NMR: (400MHz DMSO-d ₆ )δ8.44ppm(s,1H)

Synthesis of Compounds 1-3

At room temperature, compound 1-2 (22.0 g, 133 mmol) was dissolved in N, N-dimethylformamide (250 mL), and then BnBr (22.8 g, 133 mmol, 15.8 mL) and potassium carbonate (23.9 g, 173 mmol) were added. The reaction was carried out at 25°C for 1 hour. The reaction solution was poured into water (200 mL) and extracted with ethyl acetate (200 mL*3) for 3 times. The organic phases were combined, washed with saturated sodium chloride aqueous solution (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product. The crude product was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate = 100:1 to 80:1) to obtain compound 1-3.

LCMS: m/z = 251.0 [M-Cl + MeOH] ⁺

¹ H NMR: (400MHz DMSO-d ₆ )δ8.71(s,1H),7.50-7.55(m,2H),7.38-7.45(m,3H),5.18ppm(s,2H)

Synthesis of Compounds 1-4

At room temperature, compound 1-3 (27.0 g, 105 mmol) and compound 1-3A (47.8 g, 190 mmol, 53.2 mL, 50% purity) were dissolved in 270 mL of dioxane, and then compound [1,1-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane complex (8.64 g, 10.5 mmol) and potassium carbonate (32.1 g, 232 mmol) were added. The reaction was carried out at 100°C for 12 hours. The reaction solution was poured into water (200 mL) and extracted with ethyl acetate (200 mL*3) three times. The organic phases were combined, washed with saturated sodium chloride aqueous solution (300 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product. The crude product was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate = 50:1 to 1:1) to obtain compound 1-4.

LCMS: m/z = 235.0 [M + H] ⁺

¹ H NMR: (400MHz DMSO-d ₆ )δ8.66(s,1H),7.47-7.52(m,2H),7.37-7.45(m,3H),5.06(s,2H),2.42ppm(s,3H)

Synthesis of Compounds 1-5

At room temperature, compound 1-4 (11.0 g, 46.8 mmol) was dissolved in methanol (110 mL), and then Pd(dppf)Cl ₂ .CH ₂ Cl ₂ (1.91 g, 2.34 mmol) and triethylamine (11.8 g, 117 mmol, 16.3 mL) were added to the reaction solution. Carbon monoxide was replaced three times, and the reaction was carried out at 50° C. for 12 hours under 50 psi. The reaction solution was concentrated and the filtrate was obtained to obtain a crude product. The crude product was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate = 100: 1 to 50: 1) to obtain compound 1-5.

LCMS: m/z = 259.0 [M + H] ⁺

¹ H NMR: (400MHz DMSO-d ₆ )δ8.93(s,1H),7.38-7.51(m,5H),5.07(s,2H),3.92(s,3H),2.56ppm(s,3H)

Synthesis of Compounds 1-6

At room temperature, compound 1-5 (7.50 g, 29.0 mmol) was dissolved in a mixed solvent of methanol (35.0 mL) and tetrahydrofuran (35.0 mL), and then added Add compound sodium hydroxide solution (2M, 33.4mL). React at 25℃ for 1 hour. Pour the reaction solution into water (200mL), adjust pH to 3 with hydrochloric acid (2M), white solid precipitates, filter to obtain the filter cake crude product compound 1-6. The crude product is directly used for the next reaction.

LCMS: m/z = 245.1 [M + H] ⁺

¹ H NMR: (400MHz DMSO-d ₆ )δ8.86(s,1H),7.24-7.49(m,5H),5.05(s,2H),2.49ppm(s,3H)

Synthesis of Compounds 1-7

At room temperature, compound 1-6 (6.50 g, 26.6 mmol) was dissolved in ethyl acetate (20.0 mL), and wet palladium carbon (975 mg, 916 μmol, 10% purity) was added to the reaction solution. Hydrogen was replaced three times, and the reaction was carried out at 25° C. for 2 hours under 15 psi. The reaction solution was filtered and the filtrate was concentrated to obtain compound 1-7.

LCMS: m/z = 154.9 [M + H] ⁺

¹ H NMR: (400MHz DMSO-d ₆ )δ8.62(s,1H),2.46ppm(s,3H)

Synthesis of compound 2-2

At room temperature, compound 2-1 (40.0 g, 277 mmol) was dissolved in dichloromethane (400 mL), and then compound N-bromosuccinimide (49.3 g, 277 mmol) was added. The reaction was carried out at 25°C for 1 hour. The reaction solution was poured into water (80 mL) and extracted 3 times with ethyl acetate (100 mL*3). The organic phases were combined, washed with saturated sodium chloride aqueous solution (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain crude compound 2-2.

Synthesis of compound 2

At room temperature, compound 2-2 (45.0 g, 201 mmol) was dissolved in acetonitrile (450 mL), and then compound 2-3 (41.3 g, 221 mmol) and potassium carbonate (83.6 g, 605 mmol) were added. The mixture was reacted at 25°C for 1 hour. The reaction solution was filtered and the filtrate was concentrated to obtain a crude product. Compound 2 was obtained by column chromatography purification (SiO ₂ , petroleum ether: ethyl acetate = 100: 1 to 20: 1).

^1H NMR:(400MHz CHLOROFORM-d)δ4.10-4.17(m,2H),3.94(s,1H),3.23-3.49(m,4H),2.63-2.65( m,1H),2.49-2.64(m,5H),1.38(s,9H),1.18-1.21(m,3H),0.98-1.04ppm(m,3H)

Synthesis of compound 3-2

At room temperature, compound 3-1 (30.0 g, 153 mmol, 21.1 mL) was dissolved in dichloromethane (300 mL), and then compound 3-1A (17.3 g, 153 mmol, 12.2 mL) was added. The mixture was reacted at 25°C for 12 hours. The reaction solution was concentrated and the filtrate was obtained to obtain the crude product compound 3-2, which was directly used in the next step.

LCMS: m/z = 271.9 [M + H] ⁺

¹ H NMR: (400MHz DMSO-d ₆ ) δ10.15(s,1H),8.13(d,J＝8.4Hz,1H),7.98(d,J＝1.6Hz,1H),7.78(dd,J＝8.4,1.6Hz,1H),4.50ppm(s,2H)

Synthesis of compound 3

At room temperature, compound 3-2 (40.0 g, 147 mmol) was dissolved in acetone (400 mL), and potassium iodide (48.8 g, 294 mmol) was added. The mixture was reacted at 25°C for 1 hour. The reaction solution was concentrated and the filtrate was obtained to obtain a crude product compound 3-2, which can be directly used in the next step.

LCMS: m/z=363.8 (M+H)+

¹ H NMR: (400MHz DMSO-d ₆ ) δ10.15(s,1H),8.09(d,J＝8.4Hz,1H),7.97(d,J＝1.2Hz,1H),7.77(dd,J＝8.4,1.2Hz,1H),4.06ppm(s,2H)

Synthesis of compound INT A

At room temperature, compound INT 1 (9.00 g, 55.2 mmol) and compound 2 (18.1 g, 55.2 mmol) were dissolved in ethanol (90.0 mL), and then compound phosphoric acid (16.2 g, 165 mmol, 9.66 mL) was added. The mixture was reacted at 80°C for 16 hours. Then N,N-diisopropylethylamine (29.6 g, 229 mmol, 40.0 mL) and di-tert-butyl dicarbonate (18.0 g, 82.8 mmol, 19.0 mL) were added, and the mixture was reacted at 25°C for 1 hour. The reaction solution was poured into water (200 mL) and extracted three times with ethyl acetate (200 mL*3). The organic phases were combined, washed with saturated sodium chloride aqueous solution (300 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product. The crude product was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate = 100:1 to 10:1) to obtain compound INT A.

LCMS: m/z = 327.0 [M-100 + H] ⁺

¹ H NMR: (400MHz DMSO-d ₆ ) δ3.90(br d,J＝9.2Hz,2H),3.33-3.42(m,2H),2.78-2.99(m,2H),2.72(q,J＝7.6Hz,2H),2.58(br d,J＝10.4Hz,2H),1.42(s,9H),1.12-1.19ppm(m,3H)

Synthesis of compound INT B

At room temperature, compound INT A (6.00 g, 14.0 mmol) was dissolved in N, N-dimethylformamide (30.0 mL), and then compound 3 (5.10 g, 14.0 mmol) and N, N-diisopropylethylamine (5.44 g, 42.1 mmol, 7.34 mL) were added, and the mixture was reacted at 25°C for 1 hour. The reaction solution was poured into water (80.0 mL) and extracted 3 times with ethyl acetate (100 mL*3). The organic phases were combined, washed with saturated sodium chloride aqueous solution (200 mL), and dried over anhydrous sodium sulfate. The residue was dried, filtered, and the filtrate was concentrated to obtain a crude product. The crude product was purified by column chromatography (SiO ₂ , petroleum ether:ethyl acetate=100:1-5:1) to obtain compound INT B.

LCMS: m/z = 563.9 [M-100 + H] ⁺

¹ H NMR: (400MHz DMSO-d ₆ ) δ10.39(s,1H),8.12(d,J＝8.4Hz,1H),7.99-8.03(m,1H),7.77(dd,J＝8.8,1.6Hz,1H),5.35(s,2H),3.98(br d,J＝10.8Hz,2H),3.32-3.41(m,3H),2.95-3.06(m,3H),2.70(br d,J＝11.2Hz,2H),1.47(s,9H),1.16-1.22ppm(m,3H)

Synthesis of compound INT C

At room temperature, compound INT B (1.20 g, 1.81 mmol) was dissolved in dichloromethane (2.00 mL), and then trifluoroacetic acid (206 mg, 1.81 mmol, 134 μL) was added. The mixture was reacted at 25°C for half an hour. The reaction solution was concentrated to obtain the trifluoroacetate salt of the crude product compound INTC. It was directly used in the next step.

LCMS: m/z = 564.0 [M + H] ⁺

Synthesis of compound INT D

At room temperature, compound 1-7 (218 mg, 1.42 mmol) was dissolved in acetonitrile (8.0 mL), and then N-hydroxy-7-azobenzotriazole (16.0 mg, 118 μmol, 16.5 μL) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (339 mg, 1.77 mmol) were added. After reacting at 25°C for 1 hour, INT C (800 mg, 1.18 mmol, TFA) and N,N-diisopropylethylamine (763 mg, 5.91 mmol, 1.03 mL) were added, and the reaction was continued at 25°C for 1 hour. The reaction solution was poured into water (80 mL) and extracted 3 times with ethyl acetate (80 mL*3). The organic phases were combined, washed with saturated sodium chloride aqueous solution (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product. The crude product was purified by reverse phase purification (formic acid conditions: column: 330 g Flash Column Welch Ultimate XB_C18 20-40 μm; 120A; mobile phase: MeCN/H ₂ O; gradient: 10%-100% B, 60 min) to obtain compound INT D.

LCMS: m/z = 700.0 [M + H] ⁺

¹ H NMR: (400MHz DMSO-d ₆ )δ10.35(s,1H),10.24(s,1H),8.58(s,1H),8.09(d,J＝8.4Hz,1H),7.98(d,J＝1.6Hz,1H),7.73(dd,J＝8.8,1.6Hz,1H),5.32(s,2H),4.53(br d,J＝11.6Hz,1H),3.41-3.55(m,3H),3.21-3.30(m,1H),2.93-3.06(m,3H),2.82(br d,J＝11.2Hz,1H),2.60-2.69(m,1H),2.45(s,3H),1.18ppm(t,J＝7.2Hz,3H)

Example 1

Synthesis of compound 1

At room temperature, under nitrogen protection, compound INT D (50.0 mg, 71.5 μmol) was dissolved in dioxane (2 mL) and water (1 mL), and then phenylboronic acid (10.4 mg, 85.8 μmol), potassium phosphate (45.5 mg, 214 μmol) and compound [1,1-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane complex (11.6 mg, 14.3 μmol) were added, and the reaction was carried out at 80 ° C for 12 hours under nitrogen protection. Water (30 mL) was added to the reaction solution, and ethyl acetate (10 mL*3) was used for extraction. The reaction was repeated three times, and the combined organic phase was dried over sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was separated by HPLC (preparative column: C18 150×30 mm; mobile phase: [water (FA)-acetonitrile; elution gradient: 52%-82% B, 12 minutes) to obtain compound 1.

LCMS: m/z = 696.2 [M+H] ⁺

¹ H NMR: (400MHz, DMSO-d ₆ )δ10.44(s,1H),10.28-10.21(m,1H),8.59(s,1H),8.12(dd,J＝3.2,6.8Hz,2H),8.06(d,J＝8.8Hz,1H),7.98(s,1H),7.72(br d,J＝7.6Hz,1H),7.55-7.51(m,3H),5.41(s,2H),4.55(br d,J＝11.2Hz,1H),3.62-3.44(m,3H),3.29-3.23(m,1H),3.11-2.96(m,3H) ,2.89-2.81(m,1H),2.70-2.65(m,1H),2.46(s,3H),1.22(t,J＝7.6Hz,3H)

Example 2

Synthesis of compound 2

Compound INT D (50.0 mg, 71.5 umol), pyridine boronic acid (13.1 mg, 107 umol), potassium phosphate (45.5 mg, 214 umol) and 1,1-bis(diphenylphosphino)ferrocene dichloropalladium (II) dichloromethane complex (5.84 mg, 7.15 umol) were dissolved in dioxane (2 mL) and water (1 mL), and the gas was replaced by nitrogen three times. The reaction mixture was reacted at 80°C in a nitrogen atmosphere for 12 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by HPLC separation (preparative column: C18 150×30 mm; mobile phase: [water (FA)-ACN]; elution gradient: 52%-82% B, 12 minutes) to obtain compound 2.

LCMS: m/z = 697.2 [M+H] ⁺

¹ H NMR: (400MHz, DMSO-d ₆ ) δ10.43 (br s, 1H), 8.75 (br d, J = 5.6Hz, 2H), 8.58 (s, 1H), 8.08-7.95 (m, 4H), 7.71 (br d,J＝8.8Hz,1H),5.41(s,2H),4.54(br d,J＝12.0Hz,1H),3.54-3.48(m,4H),3.08-2.97(m,3H),2.85(br d,J＝9.6Hz,1H),2.67(br d,J＝10.0Hz,1H),2.54(s,1H),2.44(s,3H),1.22(br d,J＝6.8Hz,3H).

Example 3

Synthesis of compound INT 3-2

At 25°C, compound INT 3-1 (1.00 g, 4.26 mmol, HCl) was dissolved in a mixed solvent of methanol (10.0 mL) and water (1.0 mL), and then paraformaldehyde (692 mg, 8.53 mmol, 634 μL, 37% purity) and sodium cyanoborohydride (401 mg, 6.40 mmol) were added. The reaction was carried out at 25°C for 8 hours. The reaction solution was poured into water (10.0 mL), extracted 3 times with ethyl acetate (10 mL*3), and the organic phase was concentrated and filtrate was combined to obtain the crude compound INT 3-2.

LCMS: m/z = 211.9 [M + H] ⁺

¹ H NMR: (400MHz CDCl ₃ ) δ = 7.43-7.36 (m, 2H), 7.12 (d, J = 8.4Hz, 1H), 4.07 (d, J = 12.8Hz, 4H), 2.72 (s, 3H)

Synthesis of compound INT 3

At room temperature, compound INT 3-2 (900 mg, 4.24 mmol) was dissolved in dioxane (10.0 mL), and then pinacol borate (1.62 g, 6.37 mmol), potassium acetate (832 mg, 8.49 mmol) and [1,1-bis(diphenylphosphino)ferrocene] palladium dichloride (310 mg, 424 μmol) were added, and then nitrogen was replaced and reacted at 100° C. for 4 hours. The reaction solution was filtered and directly concentrated and dried under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate = 100/1 to 10/1) to obtain compound INT 3.

LCMS: m/z = 260.1 [M + H] ⁺

¹ H NMR: (400MHz CDCl ₃ ) δ = 7.69-7.63 (m, 2H), 7.21 (d, J = 7.6Hz, 1H), 3.92 (d, J = 5.2Hz, 4H), 2.59 (s, 3H), 1.35 (s, 12H)

Synthesis of compound 3

At room temperature, compound INT D (60.0 mg, 85.8 μmol) and compound INT 3 (28.9 mg, 111 μmol) were dissolved in dioxane (0.6 mL) and water (0.25 mL), and then compound [1,1-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane complex (7.01 mg, 8.59 μmol) and potassium phosphate (54.6 mg, 257 μmol) were added. The reaction was carried out at 100 °C for 4 hours. The reaction solution was poured into water (5.0 mL), extracted with ethyl acetate (5 mL*3) for 3 times, and concentrated and dried under reduced pressure. Compound 3 was obtained by preparative HPLC (FA condition: column: C18 150×30 mm; mobile phase: [water (formic acid)-acetonitrile]; gradient: 25%-55% B, 7 minutes).

LCMS: m/z = 751.3 [M + H] ⁺

¹ H NMR: (400MHz DMSO-d ₆ ) δ = 10.45 (br d,J＝1.6Hz,1H),8.54(s,1H),8.24(s,1H),8.05(d,J＝8.4Hz,1H),8.00-7.93(m,3H),7.7 2(dd,J＝1.6,8.8Hz,1H),7.36(d,J＝8.0Hz,1H),5.39(s,2H),4.60-4.47(m,1H),3.86(br d,J＝6.4Hz,4H),3.61-3.45(m,7H),3.02(br d,J＝8.8Hz,3H),2.84(br d,J＝10.0Hz,1H),2.72-2.64(m,1H),2.47-2.39(m,3H),1.21(br t,J＝7.2Hz,3H)

Example 4

Synthesis of compound INT 4-2

Compound INT4-1 (500 mg, 2.73 mmol) and potassium acetate (804 mg, 8.19 mmol) were dissolved in dioxane (10 mL), and bis-naphthalene borate (832 mg, 3.28 mmol), 1,1-bis(diphenylphosphino)ferrocene (151.43 mg, 273.16 μmol), and 1,1-bis(diphenylphosphino)ferrocene palladium chloride (199 mg, 273 μmol) were added to the above reaction solution, and the reaction mixture was reacted at 90°C for 12 hours. The reaction system was cooled to 25°C, water (50 mL) was added, and ethyl acetate (20 mL*4) was used for extraction. The combined organic phase was washed once with saturated sodium chloride aqueous solution (50 ml) and dried with anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then purified by thin layer chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound INT4-2 (150 mg).

¹ H NMR: (400MHz, DMSO-d ₆ ): δ7.53 (d, J = 7.2 Hz, 1H), 7.34 (s, 1H), 7.09 (d, J = 7.2 Hz, 1H), 3.14 (s, 4H), 1.29 (s, 12H).

Synthesis of compound 4

Compound INT D (50.0 mg, 71.5 μmol), compound INT4-2 (21.4 mg, 93.0 μmol), potassium phosphate (45.5 mg, 214 μmol), [1,1-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane (5.84 mg, 7.15 μmol) were dissolved in a mixed solution of dioxane (1.50 mL) and water (0.65 mL), and the solution was purged with nitrogen three times. The reaction mixture was reacted at 80°C for 8 hours. The reaction solution was cooled to room temperature, water (5 mL) was added to the reaction solution, and then extracted with ethyl acetate (5 mL*3). The combined organic phase was washed with saturated aqueous sodium chloride solution (8 ml) and dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product. The crude product was purified by prep-HPLC (FA condition: column: Phenomenex luna C18 150*25mm*10um; mobile phase: [water(FA)-ACN]; gradient: 50%-80% B over 10 min) to obtain compound 4 (16.71 mg).

LCMS: m/z = 722.3 [M + H] ⁺

¹ H NMR: (400 MHz, DMSO-d ₆ )

δ10.42(br d,J＝1.6Hz,2H),8.56(s,1H),8.05(d,J＝8.4Hz,1H),7.96-8.02(m,2H),7.79(s ,1H),7.71(br d,J＝8.8Hz,1H),7.22(d,J＝7.6Hz,1H),5.38(s,2H),4.53(br d,J＝11.6Hz, 1H),3.46-3.58(m,3H),3.26(br d,J＝1.2Hz,1H),3.19(s,4H),3.01(br d,J＝9.6Hz,3H),2 .83(br d,J＝11.2Hz,1H),2.66(br d,J＝10.0Hz,1H),2.44(s,3H),1.20(br t,J＝7.6Hz,3H)

Example 5

Synthesis of compound INT 5-2

At room temperature, compound INT5-1 (500 mg, 1.68 mmol) was dissolved in N,N-dimethylformamide (5.00 mL), and N,N-diisopropylethylamine (543 mg, 4.21 mmol, 733 μL) and N,O-dimethylhydroxylamine hydrochloride (328 mg, 3.37 mmol) were added. The reaction mixture was heated to 60°C and reacted for 2 hours.

The reaction solution was diluted with 15 mL (5 mL*3) of ethyl acetate and extracted. The combined organic layer was washed with 45 mL (15 mL*3) of brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by thin layer chromatography (SiO ₂ , petroleum ether/ethyl acetate = 10/1) to obtain compound INT5-2 (206 mg)

LCMS: m/z = 278.2 [M+H] +

1H NMR: (400MHz, CDCl3)7.78(d,J＝8.0Hz,2H)7.38(d,J＝8.0Hz,2H)3.80(s,2H)3.37(s,3H)2.62(s,3H)1.35(s,12H)

Synthesis of compound 5

At room temperature, INT D (50.0 mg, 71.5 μmol) and compound INT5-2 (25.7 mg, 93.0 μmol) were dissolved in a solution of dioxane (4.00 mL) and water (0.80 mL), and potassium phosphate (45.5 mg, 214 μmol) and bistriphenylphosphine ferrocene palladium catalyst (5.23 mg, 7.15 μmol) were added. The reaction mixture was evacuated, the internal air was replaced with nitrogen three times, and stirred at 80°C for 12 hours. The reaction solution was diluted with water (10 mL) and extracted with 15 mL (5 mL*3) of ethyl acetate. The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product. The crude product was separated by preparative HPLC (Waters Xbridge 150*25mm*5um; mobile phase: [water(NH4HCO3)-ACN]; gradient: 22%-52% B over 15min) to give compound 5 (11.2mg).

LCMS: m/z = 769.2 [M+H] +

1H NMR: (400MHz DMSO-d6)

10.62-10.14(m,2H)8.57(s,1H)8.12-8.02(m,3H)7.97(d,J＝1.6Hz,1H)7.71(dd,J＝8.8,1.60Hz,1H) 7.48(d,J＝8.4Hz,2H)5.40(s,2H)4.54(br d,J＝12.80Hz,1H)3.80(s,2H)3.59-3.45(m,3H)3.28(s,3H)3.27(br s,1H)3.02(br d,J＝9.2Hz,3H)2.84(br d,J＝10.4Hz,1H)2.70-2.63(m,1H)2.56(s,3H)2.44(s,3H)1.21(t,J＝7.6Hz,3H)

Example 6

Synthesis of compound INT6-2

At zero degrees, compound INT6-1A (3.60 g, 48.0 mmol) was dissolved in ethyl acetate (30.0 mL) and water (60.0 mL) solution, and sodium hydroxide (4.80 g, 120 mmol) was added. After the reaction mixture was stirred for 2 hours, compound INT6-1 (10.0 g, 40.0 mmol) was added and stirred at 20°C for 12 hours. Water (120 mL) was added to the reaction system, and ethyl acetate (60 mL*3) was extracted. The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The product was purified by reverse phase high performance liquid chromatography (0.1% NH ₃ ·H ₂ O:NH ₃ ·H ₂ O condition: 330 g Flash Column Welch Ultimate XB_C18 20-40 μm; 120A; Mobile phase: MeCN/H ₂ O; Gradient: 10-100% B over 60 min) to give compound INT6-2 (1.72 g).

LCMS: m/z = 244.0 [M + H] ⁺

¹ H NMR: (400 MHz, DMSO-d ₆ )

δ10.95(s,1H),7.58(d,J＝8.4Hz,2H),7.35(d,J＝8.4Hz,2H),4.75(s,2H),1.71(s,3H)

Synthesis of compound INT6-3

At room temperature, compound INT6-2 (1.52 g, 6.23 mmol) was dissolved in ethanol (15.0 mL), and hydrochloric acid (12 M, 1.56 mL) was added, and stirred at 65°C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain a crude product compound INT6-3 (1.49 g, HCl).

LCMS: m/z = 186.9 [M-NH ₂ +H] ⁺

Synthesis of compound INT6-4

Compound INT6-3 (1.49 g, HCl), sodium hydroxide (294 mg, 7.37 mmol) and formaldehyde (658 mg, 8.11 mmol, 603 μL) were dissolved in water (25.0 mL). The reaction mixture was reacted at 40°C for 2 hours. The reaction solution was added with water (20 mL) and extracted with ethyl acetate (50 mL*3) for 3 times. The combined organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product compound INT6-4 (1.10 g).

¹ H NMR: (400 MHz, DMSO-d ₆ )

δ7.55(d,J＝8.4Hz,2H),7.30(d,J＝8.4Hz,2H),7.12(d,J＝7.6Hz,1H),6.64(d,J＝7.2Hz,1H),5.04(s,2H) Synthesis of compound INT6-5

At room temperature, compound INT6-4 (77.0 mg, 359 μmol), bis-naphthalene borate (118 mg, 467 μmol), 1,1-bis(diphenylphosphino)ferrocene palladium chloride (26.3 mg, 35.9 μmol), 1,1-bis(diphenylphosphino)ferrocene (26.3 mg, 35.9 μmol) and potassium acetate (105 mg, 1.08 mmol) were dissolved in dioxane (3.00 mL), the gas was replaced with nitrogen three times, and the reaction mixture was reacted at 90 ° C for 12 hours. The reaction mixture was concentrated under reduced pressure to remove dioxane. The reaction solution was added with water (15 mL) and extracted with ethyl acetate (15 mL*3) three times. The combined organic phase was dried over anhydrous sodium sulfate, filtered under reduced pressure, and concentrated to obtain a crude product. The crude product was concentrated using preparative separation (preparative column: Welch Ultimate XB-SiOH 250*50*10 um; mobile phase: [Hexane-EtOH (0.1% NH ₃ ·H ₂ O)]; gradient: 1%-10% B over 15 min) to give compound INT6-5 (38.0 mg).

LCMS: m/z = 262.1 [M + H] ⁺

¹ H NMR: (400 MHz, DMSO-d ₆ )

δ7.66(br d,J＝8.0Hz,2H),7.34(br d,J＝8.0Hz,2H),7.13(d,J＝7.6Hz,1H),6.63(d,J＝7.6Hz,1H),5.09(s,2H),1.29(s,12H)

Synthesis of compound 6

At room temperature, INT D (50.0 mg, 71.5 μmol) and compound INT6-5 (24.2 mg, 93.0 μmol) were dissolved in a solution of dioxane (4.00 mL) and water (0.80 mL), and potassium phosphate (45.5 mg, 214 μmol) and bistriphenylphosphine ferrocene palladium catalyst (5.23 mg, 7.15 μmol) were added, and stirred at 80°C for 4 hours. The reaction mixture was concentrated under reduced pressure to remove dioxane. The reaction solution was added with water (20 mL) and extracted with ethyl acetate (30 mL*3) 3 times. The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product. The crude product was separated by preparative HPLC (preparative column: Waters Xbridge 150*25mm*5um; mobile phase: [water(NH ₄ HCO ₃ )-ACN]; gradient: 25%-55% B over 15min) to give compound 6 (4.79 mg).

LCMS: m/z = 753.2 [M + H] ⁺

¹ H NMR: (400 MHz, DMSO-d ₆ )

δ10.51-10.34(m,1H),8.56(s,1H),8.10(d,J＝8.0Hz,2H),8.05(d,J＝8.4Hz,1H),7.97(s, 1H),7.74-7.70(m,1H),7.48(d,J＝8.4Hz,2H),7.15(d,J＝7.6Hz,1H),6.66(d,J＝7.6Hz,1H ),5.39(s,2H),5.13(s,2H),4.53(br d,J＝12.4Hz,1H),3.55-3.47(m,4H),3.05-2.98(m, 3H),2.84(br d,J＝10.4Hz,1H),2.68-2.64(m,1H),2.44(s,3H),1.21(br t,J＝7.6Hz,3H)

Example 7

Synthesis of compound INT7-2

Compound INT7-1 (100 mg, 427 umol), bis-pinacol borate (1.08 g, 4.27 mmol), potassium acetate (125 mg, 1.28 mmol) and 1,1-bis(diphenylphosphino)ferrocenepalladium dichloride (II) dichloromethane complex (34.8 mg, 42.7 umol) were dissolved in dioxane (1 mL), and the gas was replaced with nitrogen three times. The reaction mixture was reacted at 80 ° C for 1 hour. Water (50 mL) was added to the reaction system, and ethyl acetate (20 mL*3) was extracted. The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by reverse phase liquid preparation (preparative column: 330g Flash Column; Welch Ultimate XB_C18 20-40μm; 120A, mobile phase: [water(FA)-ACN]; gradient: 0% B over 0min) to obtain compound INT7-2 (10.3mg).

LCMS: m/z = 200.0 [M + H] ⁺

¹ H NMR: (400MHz, DMSO-d ₆ ) δ8.01-7.97(m,2H),7.94-7.87(m,2H),3.09(s,3H)

Synthesis of compound 7

Compound INT7-2 (28.3 mg, 142 umol), INT D (60 mg, 85.9 umol), potassium phosphate (54.7 mg, 258 umol) and 1,1-bis(diphenylphosphino)ferrocene dichloropalladium (II) dichloromethane complex (14.0 mg, 17.2 umol) were dissolved in dioxane (4 mL) and water (1 mL), and the gas was replaced with nitrogen three times. The reaction mixture was reacted at 80°C for 1 hour. Water (30 mL) was added to the reaction system, and ethyl acetate (30 mL*5) was extracted. The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by prep-HPLC (preparative column: Welch Xtimate C18 150*25mm*5um; mobile phase: [water(FA)-ACN]; gradient: 35%-55% B over 10mins) to obtain compound 7 (9.62 mg).

LCMS: m/z = 773.2 [M + H] ⁺

¹ H NMR: (400 MHz, DMSO-d ₆ )

δ10.51-10.35(m,1H),8.54(s,1H),8.30(d,J＝8.4Hz,2H),8.11-8.01(m,3H), 7.97(d,J＝1.6Hz,1H),7.71(dd,J＝1.2,8.6Hz,1H),5.41(s,2H),4.59-4.47(m ,1H),4.34(s,1H),3.61-3.44(m,4H),3.10(s,3H),3.07-2.91(m,3H),2.90-2 .78(m,1H),2.66(br dd,J＝1.6,3.7Hz,2H),2.43(s,3H),1.21(t,J＝7.2Hz,3H)

Example 8

Synthesis of compound INT8-2

Dissolve 2-fluorophosphoryl acetate triethyl ester (6.54 g, 27.0 mmol, 5.48 mL) in tetrahydrofuran (50 mL), slowly add n-butyl lithium (2.50 M, 9.73 mL) at -78 ° C, and the reaction mixture reacts at -78 ° C for 30 minutes. Finally, add compound INT8-1 (5.00 g, 27.0 mmol) at -78 ° C, and the reaction mixture reacts at -78 ° C for 30 minutes. Add saturated aqueous ammonium chloride solution (100 mL), extract 3 times with ethyl acetate (100 mL*3), dry the combined organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain a crude product. Purify by column chromatography (SiO ₂ , petroleum ether: ethyl acetate = 1:0 to 10:1) to obtain compound 8-2 (4.80 g)

¹ H NMR: (400MHz, CDCl ₃ )

δ7.49(d,J＝8.4Hz,2H),7.35(d,J＝8.4Hz,2H),6.91-6.76(m,1H),4.27(q,J＝7.2Hz,2H),1.30-1.26(m,3H)

Synthesis of compound INT8-3

At room temperature, compound INT8-2 (4.40 g, 16.1 mmol) was dissolved in ethanol (40.0 mL) and water (30.0 mL) solution, and lithium hydroxide monohydrate (1.35 g, 32.2 mmol) was added. The reaction solution was replaced with nitrogen three times and stirred at 25°C for 2 hours. The reaction mixture was concentrated under reduced pressure to remove ethanol, the aqueous phase was acidified to pH = 1 with hydrochloric acid, and the reaction solution was extracted 3 times with ethyl acetate (60 mL*3). The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product INT8-3 (3.10 g).

¹ H NMR: (400MHz, CDCl ₃ )

δ7.54-7.47(m,2H),7.37(d,J＝8.4Hz,2H),7.03-6.92(m,1H)

Synthesis of compound INT8-4

At room temperature, compound INT8-3 (3.20 g, 13.0 mmol) and silver carbonate (180 mg, 652 μmol) were dissolved in N-methylpyrrolidone (40.0 mL). The mixture was replaced with nitrogen three times and then stirred at 130°C for 12 hours. The reaction solution was added with water (60 mL) and extracted with ethyl acetate (60 mL*3) three times. The combined organic phase was washed with water (50 mL*2), dried over anhydrous sodium sulfate, filtered under reduced pressure, and concentrated to obtain a crude product. The crude product was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate = 1:0 to 10:1) to obtain compound INT8-4 (1.86 g).

¹ H NMR:(400MHz,DMSO-d6)

δ7.82-7.57(m,1H),7.51(d,J＝8.4Hz,2H),7.34(d,J＝8.4Hz,2H),6.56-6.42(m,1H)

Synthesis of compound INT8-5

At room temperature, compound INT8-4 (200 mg, 994 μmol), bis-naphthalene borate (328 mg, 1.29 mmol), 1,1-bis(diphenylphosphino)ferrocene palladium chloride (72.7 mg, 99.4 μmol), 1,1-bis(diphenylphosphino)ferrocene (55.1 mg, 99.4 μmol) and potassium acetate (292 mg, 2.98 mmol) were dissolved in dioxane (8.00 mL), the gas was replaced with nitrogen three times, and the reaction mixture was reacted at 90 ° C for 12 hours. The reaction mixture was concentrated under reduced pressure to remove dioxane. The reaction solution was added to water (30 mL) and extracted with ethyl acetate (30 mL*3) three times. The combined organic phase was dried over anhydrous sodium sulfate, filtered under reduced pressure, and concentrated to obtain a crude product. The crude product was purified by thin layer chromatography (SiO ₂ , petroleum ether:ethyl acetate=1:0-10:1) to give compound INT8-5 (107 mg).

¹ H NMR: (400 MHz, DMSO-d ₆ )

δ7.84-7.66(m,1H),7.62(d,J＝8.0Hz,2H),7.38(d,J＝8.0Hz,2H),6.59-6.43(m,1H),1.28(s,12H)

Synthesis of compound 8

At room temperature, INT D (50.0 mg, 71.5 μmol) and compound INT8-5 (23.0 mg, 93.0 μmol) were dissolved in a solution of dioxane (4.00 mL) and water (0.80 mL), and potassium phosphate (45.5 mg, 214 μmol) and bistriphenylphosphine ferrocene palladium catalyst (5.23 mg, 7.15 μmol) were added. Stir at 80°C for 4 hours. The reaction mixture was concentrated under reduced pressure to remove dioxane. Water (20 mL) was added to the reaction solution, and extracted with ethyl acetate (30 mL*3) three times. The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product. The crude product was purified by preparative HPLC (preparative column: Waters Xbridge 150*25 mm*5 um; mobile phase: [water(NH ₄ HCO ₃ )-ACN]; gradient: 38%-68% B over 9 min) to obtain compound 8 (4.79 mg).

LCMS: m/z = 740.2 [M + H] ⁺

¹ H NMR: (400 MHz, DMSO-d ₆ )

δ10.67-10.15(m,1H),8.56(s,1H),8.05(br d,J＝8.4Hz,3H),7.97(s,1H),7.87(d,J ＝11.2Hz,1H),7.71(br d,J＝7.6Hz,1H),7.66(d,J＝11.6Hz,1H),7.54(br d,J＝8.0Hz, 2H),6.63-6.53(m,1H),5.39(s,2H),4.58-4.50(m,1H),3.55-3.49(m,3H),3.06-2.9 8(m,3H),2.86-2.82(m,1H),2.70-2.62(m,2H),2.44(s,3H),1.21(br t,J＝7.2Hz,3H)

Example 9

Synthesis of Compounds INT9-3 & INT10-3

At room temperature, compound INT9-1 (3.00 g, 8.75 mmol) was dissolved in ethanol (30 mL), and a solution of compound INT9-2 (1.39 g, 10.6 mmol) and potassium hydroxide (983 mg, 17.5 mmol) dissolved in ethanol (30 mL) was added dropwise, and the mixture was reacted at 80°C for 12 hours. After the reaction solution was concentrated, water (100 mL) was added, and the mixture was extracted three times with ethyl acetate (30 mL*3). The combined organic phase was dried over sodium sulfate, filtered and concentrated to obtain a mixture of compounds INT9-3 and INT10-3 (2.46 g,).

LCMS: m/z = 286.0 [M + H] ⁺

¹ H NMR: (400MHz, CDCl ₃ )

δ7.39-7.35(m,2H),7.32(s,1H),7.23(d,J＝1.6Hz,1H),7.04(d,J＝8.4Hz,1H),6.95(d,J＝8.4Hz ,1H),5.00(d,J＝8.4Hz,4H),4.73(d,J＝12.8Hz,4H),4.27(q,J＝7.2Hz,4H),1.33(t,J＝7.2Hz,6H)

Synthesis of Compounds INT9-4 & INT10-4

At room temperature, compound INT9-3 & INT10-3 (1.90 g, 6.64 mmol) was dissolved in ethanol (20 mL) and water (4 mL), and potassium hydroxide (788 mg, 14.0 mmol) was added, and the mixture was reacted at 80°C for 12 hours. Dilute hydrochloric acid (30 mL, 1 M) was added to the reaction system, and the mixture was extracted three times with ethyl acetate (10 mL*3). The aqueous phase was adjusted to pH 10 with sodium carbonate, and then extracted four times with ethyl acetate (10 mL*4). The combined organic phase was dried over sodium sulfate, filtered and concentrated to obtain a mixture of compounds INT9-4 and INT10-4 (0.669 g, 3.13 mmol, 47.0% yield).

LCMS: m/z = 214.0 [M+H] ⁺

Synthesis of Compounds INT9-5 & INT10-5

At room temperature, a mixture of compounds INT9-4 and INT10-4 (400 mg, 1.87 mmol, 1 eq) was dissolved in formic acid (350 mg, 7.30 mmol, 5 mL), and then a compound formaldehyde aqueous solution (303 mg, 3.74 mmol, 278 μL, 37% purity) was added, and the mixture was reacted at 120°C for 12 hours. The reaction solution was concentrated to obtain a crude product, which was prepared by reverse phase high performance liquid chromatography (preparative column: 330 g Flash Column Welch Ultimate XB_C18 20-40 μm; 120A; mobile phase: [water (FA) -ACN]; gradient: 0% -0% B over 90 mins) to obtain a mixture of compounds INT9-5 and INT10-5 (280 mg,).

LCMS: m/z = 228.0 [M + H] ⁺

¹ H NMR: (400MHz, CDCl ₃ )

δ7.24(td,J＝2.8,8.0Hz,2H),7.16(s,1H),7.12(s,1H),6.88(d,J＝8.0Hz,1H),6.84(d,J＝8.0Hz,1H),3.84-3.61(m,4H),2.72(d,J＝1.2Hz,6H)

Isolation and purification of compound INT9-5 and compound INT10-5

A mixture of compounds INT9-5 and INT10-5 (530 mg, 2.32 mmol, 1 eq) was separated by SFC (preparative column: DAICEL CHIRALPAK IG (250 mm*50 mm, 10 um); mobile phase: [CO ₂ -EtOH (0.1% NH ₃ ·H ₂ O)]; B%: 15%, isocratic elution mode) to obtain compounds INT9-5 (198 mg,) and INT10-5 (202 mg).

Compound INT9-5

LCMS: m/z = 228.0 [M + H] ⁺

¹ H NMR: (400MHz, CDCl ₃ )

δ7.24(dd,J＝2.0,8.0Hz,1H),7.13(s,1H),6.95-6.84(m,1H),5.20-4.52(m,2H),3.72(br s,2H),2.72(s,3H)

SFC: retention time: RT = 0.911 min

Column: Chiralpak AD-3 50×4.6mm ID, 3um; mobile phase: Phase A for CO ₂ , and Phase B for EtOH (0.05% DEA); Gradient elution: EtOH (0.05% DEA) in CO ₂ from 5% to 40%; Flow rate: 3mL/min; Detector: PDA; Column Temp: 35C; Back Pressure:100Bar;

Compound INT10-5

LCMS: m/z = 228.0 [M + H] ⁺

¹ H NMR: (400MHz, CDCl ₃ )

δ7.24(dd,J＝2.0,8.0Hz,1H),7.18-7.13(m,1H),6.84(d,J＝8.0Hz,1H),5.05-4.53(m,2H),3.83-3.66(m,2H),2.72(s,3H)

SFC: retention time: RT = 1.064 min

Column: Chiralpak AD-3 50×4.6mm ID, 3um; mobile phase: Phase A for CO ₂ , and Phase B for EtOH (0.05% DEA); Gradient elution: EtOH (0.05% DEA) in CO ₂ from 5% to 40%; Flow rate: 3mL/min; Detector: PDA; Column Temp: 35C; Back Pressure:100Bar;

Synthesis of compound INT9-6

At room temperature, compound INT9-5 (100 mg, 438 μmo) and bis-naphthalene borate (222 mg, 876 μmol) were dissolved in 1,4-dioxane (2 mL), and then 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (32.0 mg, 43.8 μmol) and sodium carbonate (139 mg, 1.32 mmol) were added. The reaction system was evacuated and replaced with nitrogen three times, and then stirred at 80 ° C for 12 hours. Water (30 mL) was added to the reaction system, and ethyl acetate (10 mL*3) was extracted three times. The combined organic phase was dried over sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was purified by reverse phase high performance liquid chromatography (preparative column: 330 g Flash Column Welch Ultimate XB_C18 20-40 μm; 120A; mobile phase: [water(FA)-ACN]; gradient: 0% -0% B over 90 mins) to give compound INT9-6 (91.0 mg).

LCMS: m/z = 276.1 [M + H] ⁺

¹ H NMR: (400MHz, CDCl ₃ )

δ7.55(d,J＝7.6Hz,1H),7.43(s,1H),7.02(d,J＝7.6Hz,1H),5.27-4.55(m,2H),3.81(br s,2H),2.73(s,3H),1.27(s,12H)

Synthesis of compound 9

Compound INT D (50.0 mg, 71.5 μmol), compound INT9-6 (21.6 mg, 78.7 μmol), potassium phosphate (45.5 mg, 214 μmol) and 1,1-bis(diphenylphosphino)ferrocenepalladium dichloride (5.23 mg, 7.15 μmol) were dissolved in dioxane (2 mL) and water (0.5 mL), and the gas was replaced with nitrogen three times. The reaction mixture was reacted at 80°C for 1 hour. Water (30 mL) was added to the reaction system, and ethyl acetate (10 mL*3) was extracted. The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by prep-HPLC (preparative column: Waters Xbridge 150*25 mm*5 um; mobile phase: [water(NH ₄ HCO ₃ )-ACN]; gradient: 18%-48% B over 15 min) to obtain compound 9 (5.04 mg).

LCMS: m/z = 767.3 [M + H] ⁺

¹ H NMR: (400 MHz, DMSO-d ₆ )

δ10.59-10.25(m,1H),8.54(br s,1H),8.07(d,J＝8.5Hz,1H),8.00-7.92(m,2H),7.86(s,1H ),7.72(dd,J＝1.4,8.6Hz,1H),7.30(d,J＝8.3Hz,1H),5.40(s,2H),4.96(br s,2H),4.54(br d,J＝11.9Hz,1H),4.10-3.70(m,2H),3.61-3.47(m,3H),3.28-3.20(m,2H),3.12-2.92(m,3 H),2.91-2.79(m,1H),2.72(s,3H),2.69-2.61(m,1H),2.44(s,3H),1.21(br t,J＝7.4Hz,3H)

Synthesis of Compound INT10-6

At room temperature, compound INT10-5 (100 mg, 438 μmol) and bis-naphthalene borate (222 mg, 876 μmol) were dissolved in 1,4-dioxane (2 mL), and 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (32.0 mg, 43.8 μmol) and sodium carbonate (139 mg, 1.32 mmol) were added. The reaction system was evacuated and replaced with nitrogen three times, and stirred at 80 ° C for 12 hours. Water (30 mL) was added to the reaction system, and ethyl acetate (10 mL*3) was extracted three times. The combined organic phase was dried over sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was purified by reverse phase high performance liquid chromatography (preparative column: 330 g Flash Column Welch Ultimate XB_C18 20-40 μm; 120A; mobile phase: [water(FA)-ACN]; gradient: 10% -100% B over 60 mins) to give compound INT10-6 (82.0 mg).

LCMS: m/z = 276.1 [M + H] ⁺

¹ H NMR: (400MHz, CDCl ₃ )δ7.57(d,J＝7.6Hz,1H),7.47(s,1H),6.98(d,J＝7.6Hz,1H),5.18-4.67(m,2H),3.89-3.78(m,2H),2.75(s,3H),1.28(s,12H)

Synthesis of compound 10

Compound INT D (50 mg, 71.5 μmol), compound INT10-6 (22 mg, 79.9 μmol), potassium phosphate (45.5 mg, 214 μmol) and 1,1-bis(diphenylphosphino)ferrocenepalladium dichloride (5.23 mg, 7.15 μmol) were dissolved in dioxane (2 mL) and water (0.5 mL), and the gas was replaced with nitrogen three times. The reaction mixture was reacted at 80°C for 1 hour. Water (30 mL) was added to the reaction system, and ethyl acetate (10 mL*3) was extracted. The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by prep-HPLC (preparative column: Waters Xbridge 150*25 mm*5 um; mobile phase: [water(NH ₄ HCO ₃ )-ACN]; gradient: 18%-48% B over 15 min) to obtain compound 10 (2.76 mg).

LCMS: m/z = 767.3 [M + H] ⁺

¹ H NMR: (400 MHz, DMSO-d ₆ )

δ10.51-10.31(m,1H),8.54(s,1H),8.06(d,J＝8.4Hz,1H),7.99-7.92(m,2H),7.89(s,1H),7.72(br d,J＝8.4Hz,1H),7.25(d,J＝8.0Hz,1H),5.45-5.35(m,2H),5.06-4.80(m,2H),4.60-4.48(m,1H),4.13-3.68(m,2H), 3.60-3.44(m,3H),3.28-3.17(m,2H),3.10-2.93(m,3H),2.89-2.78(m,1H),2.71-2.70(m,1H),2.71(s,2H),2.67(br s,1H),2.43(s,3H),1.21(br t,J＝7.2Hz,3H)

Test Example 1: In vitro activity test

1.1. Reagents, consumables and instruments are shown in Table 1

Table 1 Reagents, consumables and instruments used in the test

1.2. SW48 cell antiproliferation assay

SW48 cells were seeded in a white 384-well plate, with 40 μL of cell suspension per well, containing 500 SW48 cells. The cell plate was placed in a carbon dioxide incubator for overnight culture. The compound to be tested was diluted 3 times to the 10th concentration, i.e., from 2000 μM to 101.61 nM, using a dispenser, and a double-well experiment was set up. 78 μL of culture medium was added to the middle plate, and then 2 μL of the gradient dilution compound per well was transferred to the middle plate according to the corresponding position, and 10 μL of each well was transferred to the cell plate after mixing. The concentration range of the compound transferred to the cell plate was 10 μM to 0.51 nM. The cell plate was placed in a carbon dioxide incubator for 6 days. Prepare another cell plate, and read the signal value on the day of drug addition as the maximum value (Max value in the equation below) for data analysis. Add 10 μL of cell viability chemiluminescent detection reagent to each well of this cell plate and incubate at room temperature for 10 minutes to stabilize the luminescent signal. Read using a multi-label analyzer. After the cell plate with the compound added is incubated, 10 μL of cell viability chemiluminescent detection reagent is added to each well of the cell plate, and the plate is incubated at room temperature for 10 minutes to stabilize the luminescent signal. The reading is performed using a multi-label analyzer.

Data Analysis:

The raw data were converted into inhibition rate using the equation (Sample-Min)/(Max-Min)*100%, and the _IC50 value was obtained by four-parameter curve fitting (obtained using the "log(inhibitor)vs.response--Variable slope" mode in GraphPad Prism).

1.3. The results of in vitro activity test are shown in Table 2

Table 2 Antiproliferative activity of the tested compounds on SW48 cells

1.4. Experimental conclusion

The compound of the present application has excellent WRN enzyme inhibitory activity and good anti-proliferation activity against MSI type tumor cells.

Claims (11)

A compound of formula (I) or a pharmaceutically acceptable salt thereof,
in, Ring A is selected from 5-6 membered heteroaryl, benzene ring and 5-6 membered heterocycloalkenyl; L ₁ is selected from Each R ₁ is independently selected from H, C _1-3 alkyl, C _2-3 alkenyl, C _2-3 alkynyl, or two R ₁ on adjacent carbon atoms together with the carbon atom to which they are connected form a 3-10 membered ring, or two R ₁ on two non-adjacent carbon atoms form a bridge containing 1 or 2 carbon atoms, the C _1-3 alkyl group is optionally substituted by 1, 2, 3 or 4 R _a , the 3-10 membered ring, C _2-3 alkenyl, C _2-3 alkynyl, and the 3-10 membered ring is optionally substituted with 1, 2, 3 or 4 R _b ; each R ₂ is independently selected from H, F, Cl, Br, NH ₂ , CN, OH and C _1-3 alkyl, wherein the C _1-3 alkyl is optionally substituted with 1, 2, 3 or 4 R _c ; _R3 is selected from _C1-3 alkyl; R ₄ is selected from R _a is selected from R _b is selected from H, F, Cl, Br, NH ₂ , CN, OH and C _1-3 alkyl; R _c is selected from H, F, Cl, Br, NH ₂ , CN, OH and C _1-3 alkyl; n is selected from 0, 1, 2, 3 and 4; m is selected from 0, 1, 2, 3 and 4; The condition is, When ring A is selected from 5-6 membered heterocycloalkenyl, _L1 is selected from When R ₄ is not The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein R _b is selected from H, F, Cl, Br, NH ₂ , CN, OH, CH ₃ , CH ₂ CH ₃ and CH ₂ CH ₂ CH ₃ . The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein R _c is selected from H, F, Cl, Br, NH ₂ , CN, OH, CH ₃ , CH ₂ CH ₃ and CH ₂ CH ₂ CH ₃ . The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein ring A is selected from The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein each R ₁ is independently selected from H, CH ₃ , CH ₂ CH ₃ , CH ₂ ＝CH ₂ , HC≡CH, Or two R ₁ on two adjacent carbon atoms together with the carbon atom to which they are attached form The CH ₃ and CH ₂ CH ₃ are optionally substituted by 1, 2, 3 or 4 _Ra , and the CH ₂ ＝CH ₂ , HC≡CH, Optionally substituted with 1, 2, 3 or 4 R _b . The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the structural unit Selected from The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein R ₃ is selected from CH ₃ , CH ₂ CH ₃ and CH ₂ CH ₂ CH ₃ . The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from
wherein ring A, R ₁ , R ₂ , R ₃ , R ₄ , L ₁ , n and m are as defined in any one of claims 1-7. The following compound or a pharmaceutically acceptable salt thereof,

Use of the compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating a disease associated with WRN. The use according to claim 10, wherein the disease associated with WRN refers to cancer with high microsatellite instability (MSI-H) or mismatch repair deficiency (dMMR), further preferably colorectal cancer, gastric cancer, endometrial cancer, lung cancer, head and neck cancer, liver cancer, breast cancer, melanoma, and renal cell carcinoma.