CN117624194A - KRAS G12D inhibitor and application thereof in medicine - Google Patents

Abstract

The present invention relates to the compound of formula (I), its stereoisomer, tautomer, deuterated product or pharmaceutical salt, its preparation method, pharmaceutical composition containing the compound, and its use as a treatment and/or Application of drugs to prevent diseases mediated by KRAS.

Description

KRAS G12D inhibitors and their medical applications

Technical Field

The present invention relates to the field of medical technology, and in particular to a compound of formula (I), its stereoisomers, tautomers, deuterated substances or pharmaceutically acceptable salts, a preparation method thereof, a pharmaceutical composition containing the compound, and its use as a drug for treating cancer.

Background Art

Clinical data show that RAS is the gene with the highest mutation rate in human tumors. About 20-30% of all tumors have RAS mutations, and about 98% of pancreatic cancers, 52% of colon cancers, 43% of multiple myeloma, and 32% of lung adenocarcinomas have RAS gene mutations. The most common mutation of RAS is point mutation, which often occurs at codons 12, 13, and 61, and the most common mutation is at codon 12, such as G12C, G12D, or G12V.

Currently, drug development targeting KRAS mutations is one of the current hot spots in new drug research. KRAS G12C inhibitors AMG510 (WO2018217651A1) and MRTX849 (WO2019099524A1) have entered the late clinical stage; and MIRATI is leading in the development of G12D inhibitors (WO2021041671A1).

Based on the importance of abnormal KRAS activation in cancer progression and the prevalence of KRAS gene mutations in human cancers, KRAS has always been a target of interest for drug developers. Although progress has been made in this field, there is still a need for improved KRAS G12D mutant protein inhibitors in the art.

Summary of the invention

The technical problem to be solved by the present invention is to overcome the problem of lack of KRAS G12D mutant protein inhibitors in the prior art, and to provide a compound represented by general formula (I) and its application. The compound represented by general formula (I) provided by the present invention has a good inhibitory effect on KRAS G12D mutant protein.

The present invention provides a compound represented by general formula (I), its stereoisomers, tautomers, deuterated substances or pharmaceutically acceptable salts:

in,

X is selected from a bond, NH, O or S;

is selected from a single bond or a double bond;

X ₁ is selected from O, (CH ₂ ) _0-3 , C(R ₄ ) _1-2 , C(O) or NR ₄ ; R ₄ are each independently selected from H, halogen, C _1-6 alkyl, substituted C _1-6 alkyl, C _1-6 alkoxy, C _3-6 cycloalkyl or substituted C _3-6 cycloalkyl;

_X2 is selected from _CR5 or N; _R5 is selected from absent or H;

X ₃ is selected from O, (CH ₂ ) _0-3 , C(O), C(R ₆ ) _1-2 or NR ₆ ; R ₆ are each independently selected from H, amino, substituted amino, cyano, C _1-6 alkyl, substituted C _1-6 alkyl, halogen, C _2-6 alkenyl, substituted C _2-6 alkenyl, C _3-6 cycloalkyl or substituted C _3-6 cycloalkyl;

X ₄ is selected from O, NR ₇ , (CH ₂ ) _0-3 , C(R ₇ ) _1-2 or C(O); R ₇ are each independently selected from H, amino, substituted amino, cyano, C _1-6 alkyl, substituted C _1-6 alkyl, C _1-6 alkoxy, C _3-6 cycloalkyl-O-, halogen, C _2-6 alkenyl, substituted C _2-6 alkenyl, C _3-6 cycloalkyl or substituted C _3-6 cycloalkyl;

L is selected from a bond, -C _1-3 alkylene-, -C _3-14 cycloalkyl _- C _0-3 alkylene- or -3-14 membered heterocyclyl-C 0-3 alkylene-, wherein the -C _1-3 alkylene-, -C _3-14 cycloalkyl-C _0-3 alkylene- or -3-14 membered heterocyclyl-C _0-3 alkylene- is optionally further substituted with one or more _Ra ;

The ring A is selected from C _5-14 cycloalkyl, 5-14 membered heterocyclyl, C _6-18 aryl or 5-18 membered heteroaryl, the C _5-14 cycloalkyl or 5-14 membered heterocyclyl is selected from a monocyclic ring, a condensed ring, a spirocyclic ring or a bridged ring, and the C _5-14 cycloalkyl, 5-14 membered heterocyclyl, C _6-18 aryl or 5-18 membered heteroaryl is optionally further substituted by one or more _Ra ;

Selected from in, is a double bond, is selected from Z configuration or E configuration; R ₈ is each independently selected from H, halogen, hydroxyl, cyano, C _1-6 alkyl, C _1-6 haloalkyl, C _3-14 cycloalkyl, 3-14 membered heterocyclyl, C _6-18 aryl or 5-18 membered heteroaryl, wherein the C _1-6 alkyl, C _1-6 haloalkyl, C _3-14 cycloalkyl, 3-14 membered heterocyclyl, C _6-18 aryl or 5-18 membered heteroaryl is optionally further substituted with one or more halogen, hydroxyl, cyano, carboxyl, amino, nitro, C _1-6 alkyl or C _1-6 alkoxy;

R ₂ is selected from C _3-14 cycloalkyl, 3-14 membered heterocyclyl, C _6-18 aryl or 5-18 membered heteroaryl, wherein the C _3-14 cycloalkyl, 3-14 membered heterocyclyl, C _6-18 aryl or 5-18 membered heteroaryl is optionally further substituted with one or more _Ra ;

_Ra is independently selected from H, hydroxy, cyano, halogen, _C1-6 alkyl, _C1-6 haloalkyl, _C1-6 alkoxy, _-C0-3 alkylene- _ORb , -OC(=O) _C1-6 alkyl, -C0-3 alkylene- _SRb , _-C0-3 alkylene-N( _Rb ) ₂ , _-C0-3 alkylene-S(=O) _Rb , _{-C0-3 alkylene-S(=O)2Rb, -C0-3 alkylene - SRb , -C0-3 alkylene} -S( _Rb ) ₅ , _-C0-3 alkylene-C(=O) _Rb , _-C0-3 alkylene-C(=O) _ORb , _-C0-3 alkylene-C(=O)N( _Rb ) ₂ , _C2-6 alkenyl, _C2-6 alkynyl, -C -C _0-3 alkylene-C _3-14 cycloalkyl, -C _0-3 alkylene-(3-14 membered heterocyclyl), -C _0-3 alkylene-C _6-18 aryl or -C _0-3 alkylene-(5-18 membered heteroaryl), the C _1-6 alkyl, C _{1-6 haloalkyl, C 1-6 alkoxy} , C _2-6 alkenyl, C _2-6 alkynyl, -C _0-3 alkylene-C 3-14 cycloalkyl, -C _0-3 alkylene-( _3-14 membered heterocyclyl), -C _0-3 alkylene-C _6-18 aryl or -C _0-3 alkylene-(5-18 membered heteroaryl) is optionally further substituted with one or more R _b ;

each R _b is independently H, halogen, hydroxy, cyano, C _1-6 alkyl, C _3-6 cycloalkyl, C _{1-6 haloalkyl, C 3-14 cycloalkyl} , 3-14 membered heterocyclyl, C _6-18 aryl or 5-18 membered heteroaryl, said C _1-6 alkyl, C _3-6 cycloalkyl, C _1-6 haloalkyl, C _3-14 cycloalkyl, 3-14 membered heterocyclyl, C _6-18 aryl or 5-18 membered heteroaryl optionally further substituted with one or more halogen, C _1-6 alkyl or C _1-6 haloalkyl;

R ₃ is selected from H, halogen, -C _0-3 alkylene cyano, C _1-6 alkyl, C _1-6 alkoxy or C _1-6 haloalkyl.

In some embodiments, the compound represented by formula (I), its stereoisomers, tautomers, deuterated substances or pharmaceutically acceptable salts are selected from the compounds represented by formula (IA) to formula (IF), their stereoisomers, tautomers, deuterated substances or pharmaceutically acceptable salts:

wherein R ₁ , R ₂ , R ₃ , R ₄ , R ₆ , R ₇ , X, L and A have the same definitions as in formula (I).

A compound represented by general formula (I), its stereoisomers, tautomers, deuterated substances or pharmaceutically acceptable salts, which is selected from compounds represented by formula (IA-1) to formula (IF-1), their stereoisomers, tautomers, deuterated substances or pharmaceutically acceptable salts:

wherein R ₁ , R ₂ , R ₃ , R ₄ , R ₆ , R ₇ and A have the same definitions as in formula (I).

In some embodiments, Ring A in Formula (I) is selected from C _5-14 cycloalkyl or 5-14 membered heterocyclyl.

In some embodiments, the 5-14 membered heterocyclic group in formula (I) is a condensed ring.

In some embodiments, the fused ring in formula (I) is selected from

In some embodiments, in formula (I) Selected from

In some embodiments, _R8 in formula (I) is independently selected from H, halogen, _C1-6 alkyl, _C3-14 cycloalkyl, 3-14 membered heterocyclyl, _C6-18 aryl or 5-18 membered heteroaryl, and the _C3-14 cycloalkyl, 3-14 membered heterocyclyl, _C6-18 aryl or 5-18 membered heteroaryl is optionally further substituted with one or more halogen, hydroxyl, cyano, carboxyl, amino, nitro or _C1-6 alkyl.

In some embodiments, in formula (I) Selected from

In some embodiments, in formula (I) Selected from

In some embodiments, the compound represented by formula (I), its stereoisomer, tautomer, deuterated substance or pharmaceutically acceptable salt is selected from the compounds represented by formula (IA-1-1) to formula (IF-1-1), its stereoisomer, tautomer, deuterated substance or pharmaceutically acceptable salt:

wherein R ₁ , R ₂ , R ₃ , R ₄ , R ₆ and R ₇ have the same definitions as in formula (I).

In some embodiments, R ₄ in formula (I) is H or halogen, and the halogen is preferably F.

In some embodiments, _R6 in formula (I) is H, alkyl or halogen.

In some embodiments, _R7 in formula (I) is H, _C1-6 alkoxy or _C3-6 cycloalkyl-O-.

In some embodiments, _R2 in formula (I) is selected from _C6-18 aryl or 5-18 membered heteroaryl, and the _C6-18 aryl or 5-18 membered heteroaryl is optionally further substituted by one or more _Ra , and _Ra is independently selected from H, hydroxyl, cyano, amino, halogen, _C1-6 alkyl, _C1-6 alkoxy, _C2-6 alkenyl, _C2-6 alkynyl or _-C0-3 alkylene- _C3-14 cycloalkyl.

In some embodiments, _R2 in formula (I) is selected from Said Optionally further substituted _with one or more _Ra independently selected from H, hydroxy, cyano, halogen, amino, _C1-6 alkyl, _C1-6 alkoxy, _C2-6 alkenyl, _C2-6 alkynyl or _-C0-3 alkylene- _C3-14 cycloalkyl.

In some embodiments, _R2 in formula (I) is selected from

In some embodiments, R ₃ in formula (I) is H, methoxy or halogen.

In some embodiments, formula (I) is selected from

The present invention also provides a pharmaceutical composition, wherein the pharmaceutical composition comprises a therapeutically effective amount of at least one compound represented by formula (I), a stereoisomer, a tautomer or a pharmaceutically acceptable salt thereof.

The present invention provides the use of a compound represented by structural formula (I) or a pharmaceutical composition thereof in the preparation of a drug.

The present invention further provides a preferred technical solution for the application:

Preferably, the application is application in the preparation of drugs for treating and/or preventing cancer.

Preferably, the use is for preparing a drug for treating a disease mediated by KRAS G12D. Preferably, the disease is cancer.

Preferably, the cancer is selected from breast cancer, multiple myeloma, bladder cancer, endometrial cancer, gastric cancer, cervical cancer, rhabdomyosarcoma, non-small cell lung cancer, small cell lung cancer, pleomorphic lung cancer, ovarian cancer, esophageal cancer, melanoma, colorectal cancer, hepatoma, head and neck tumors, hepatobiliary carcinoma, myelodysplastic syndrome, malignant glioma, prostate cancer, thyroid cancer, Schwann cell tumor, squamous cell carcinoma of the lung, lichenoid keratosis, synovial sarcoma, skin cancer, pancreatic cancer, testicular cancer or liposarcoma.

The present invention also provides a method for treating and/or preventing a disease, comprising administering to a subject a therapeutically effective amount of at least one compound represented by structural formula (I) or a pharmaceutical composition containing the compound.

The present invention also provides a method for treating and/or preventing a disease mediated by KRAS G12D, comprising administering to a subject a therapeutically effective amount of at least one compound represented by structural formula (I) or a pharmaceutical composition containing the same.

The present invention also provides a method for treating cancer, comprising administering to a subject a therapeutically effective amount of at least one compound represented by structural formula (I) or a pharmaceutical composition containing the compound.

Preferably, in the above method, the KRAS G12D-mediated disease is cancer.

Preferably, in the above method, the cancer is selected from breast cancer, multiple myeloma, bladder cancer, endometrial cancer, gastric cancer, cervical cancer, rhabdomyosarcoma, non-small cell lung cancer, small cell lung cancer, pleomorphic lung cancer, ovarian cancer, esophageal cancer, melanoma, colorectal cancer, hepatoma, head and neck tumors, hepatobiliary cell carcinoma, myelodysplastic syndrome, malignant glioma, prostate cancer, thyroid cancer, Schwann cell tumor, squamous cell carcinoma of the lung, lichenoid keratosis, synovial sarcoma, skin cancer, pancreatic cancer, testicular cancer or liposarcoma.

Unless otherwise indicated, general chemical terms used in the structural formulae have their usual meanings.

For example, the term "halogen," as used herein, refers to fluorine, chlorine, bromine, or iodine, unless otherwise indicated.

In the present invention, unless otherwise specified, "alkyl" includes a linear or branched monovalent saturated hydrocarbon group. For example, alkyl includes methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 3-(2-methyl)butyl, 2-pentyl, 2-methylbutyl, neopentyl, n-hexyl, 2-hexyl, 2-methylpentyl, etc. Similarly, " _1-6 " in " _C1-6 alkyl" refers to a group containing 1, 2, 3, 4, 5 or 6 carbon atoms in a linear or branched form.

"Alkoxy" refers to the oxygen ether form of the aforementioned straight-chain or branched alkyl groups, ie, -O-alkyl.

The term "alkylene" refers to a divalent alkyl linking group. Alkylene formally refers to an alkane with two CH bonds replaced as the point of attachment of the alkylene to the rest of the compound. Similarly, the "C _1-3 " in C _1-3 alkylene refers to an alkylene containing 1, 2, or 3 carbon atoms, including but not limited to methylene, 1,2-ethylene, 1,3-propylene, or 1,2-isopropylene.

The term "haloalkyl" refers to an alkyl group in which one or more H has been replaced by a halogen atom.

The term "oxo" or "oxo group" refers to an oxygen atom in the form of a divalent substituent, which forms a carbonyl group when attached to C, and forms a sulfoxide or sulfone group or an N-oxide group when attached to a heteroatom.

In the present invention, unless otherwise specified, the term "aromatic ring", "aromatic ring" or "aromatic heterocycle" refers to a polyunsaturated carbon ring or heterocycle with aromatic characteristics (having (4n+2) delocalized π electrons, where n is an integer).

The term "aryl", in the present invention, unless otherwise specified, refers to an unsubstituted or substituted monocyclic or condensed aromatic group including carbon ring atoms. Preferably, C _6-18 aryl, more preferably, aryl is a C _6-10 monocyclic or bicyclic aromatic ring group. Preferably, it is phenyl, naphthyl; most preferably, naphthyl. The aryl ring can be fused to a heteroaryl, heterocyclic or cycloalkyl group, wherein the ring connected to the parent structure is an aryl ring, non-limiting examples include but are not limited to benzocyclopentyl.

The term "heterocyclyl" refers to a ring system having at least one cyclized alkyl or cyclized alkenyl group containing a heterocyclic ring, wherein the heteroatom is selected from N, O and/or S. The heterocyclyl may include a monocyclic or polycyclic ring (e.g., having 2, 3 or 4 fused rings, spirocyclic rings, bridged rings, etc.). The heterocyclyl may be connected to the rest of the compound via a ring-forming carbon atom or a ring-forming heteroatom. Preferably, a 3-14-membered heterocyclyl is used, and the "3-14 members" in the 3-14-membered heterocyclyl refers to a heterocyclyl composed of 3-14 C, N, O or S ring atoms; more preferably, a 5-14-membered heterocyclyl and a 3-8-membered heterocyclyl, and even more preferably a 3-6-membered heterocyclyl. Wherein the nitrogen or sulfur heteroatom can be selectively oxidized, and the nitrogen heteroatom can be selectively quaternized. Examples of these heterocyclyls include, but are not limited to Azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, oxopiperazinyl, oxopiperidinyl, tetrahydrofuranyl, dioxolanyl, tetrahydroimidazolyl, tetrahydrothiazolyl, tetrahydrooxazolyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, thiomorpholinyl sulfoxide, thiomorpholinyl sulfone and tetrahydrooxadiazolyl. The heterocyclic group can be fused on an aryl, heteroaryl or cycloalkyl ring, wherein the ring connected to the parent structure is a heterocyclic group.

The term "heteroaryl", in the present invention, unless otherwise specified, refers to a monocyclic or polycyclic (e.g., 2, 3 or 4 fused rings, spiro rings, bridged rings, etc.) aromatic heterocycle having at least one heteroatom, wherein the heteroatom is selected from N, O and/or S, and wherein the nitrogen or sulfur heteroatom can be selectively oxidized, and the nitrogen heteroatom can be selectively quaternized. Preferably, it is a 5-18-membered heteroaryl, wherein the "5-18 members" in the 5-18-membered heteroaryl refers to a heteroaryl composed of 5-18 C, N, O or S ring atoms. More preferred is a 5-10-membered heteroaryl; more preferred is a 5-6-membered heteroaryl. Examples of heteroaryl include, but are not limited to, thienyl, furanyl, imidazolyl, isoxazolyl, oxazolyl, pyrazolyl, pyrrolyl, thiazolyl, thiadiazolyl, triazolyl, pyridyl, pyridazinyl, indolyl, azaindolyl, indazolyl, benzimidazolyl, benzofuranyl, benzothienyl, benzisoxazolyl, benzothiazolyl, benzothiazolyl, benzothiadiazolyl, benzotriazolyl adenine, quinolyl or isoquinolyl. The heteroaryl may be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring attached to the parent structure is a heteroaryl ring.

The term "cycloalkyl" refers to a ring system having at least one cyclized alkyl group. Preferably, C _3-14 cycloalkyl, wherein "C _3-14 " means that the cycloalkyl group may have 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms. Cycloalkyl groups may include monocyclic and polycyclic rings (e.g., having 2, 3 or 4 fused rings, spirocyclic rings, bridged rings, etc.). In some embodiments, cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, etc.; the cycloalkyl group may also be fused to an aryl, heterocyclic or heteroaryl ring, wherein the ring connected to the parent structure is a cycloalkyl group.

The term "substituted" means that one or more hydrogen atoms in the group are replaced by the same or different substituents. Typical substituents include, but are not limited to, halogen (F, Cl, Br or I), C _1-8 alkyl, C _3-12 cycloalkyl, -OR ¹ , -SR ¹ , =O, =S, -C(O)R ¹ , -C(S)R ¹ , =NR ¹ , -C(O)OR ¹ , -C(S)OR ¹ , -NR ¹ R ² , -C(O)NR ¹ R ² , cyano, nitro, -S(O) ₂ R ¹ , -OS(O ₂ )OR ¹ , -OS(O) ₂ R ¹ , -OP(O)(OR ¹ )(OR ² ); wherein R ¹ and R ² are independently selected from -H, C _1-6 alkyl, C _1-6 haloalkyl or C _3-6 cycloalkyl. In some embodiments, substituents are independently selected from groups comprising -F, -Cl, -Br, -I, -OH, trifluoromethoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, _{-SCH3 , -SC2H5} , carbaldehyde, -C( _OCH3 ), cyano, nitro, _-CF3 , _-OCF3 , amino, dimethylamino, methylthio, sulfonyl, and acetyl.

When the number of a linking group is 0, such as -(CH ₂ ) ₀ -, it means that the linking group is a bond.

The term "pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids.

When the compound provided by the invention is an acid, its corresponding salt can be conveniently obtained from pharmaceutically acceptable nontoxic bases, including inorganic bases and organic bases. Salts derived from inorganic bases include salts of aluminum, ammonium, calcium, copper (high and low valence), ferric iron, ferrous iron, lithium, magnesium, manganese (high and low valence), potassium, sodium, zinc and the like. Particularly preferred are salts of ammonium, calcium, magnesium, potassium and sodium. Nontoxic organic bases that can be derived into pharmaceutically acceptable salts include primary amines, secondary amines and tertiary amines, also include cyclic amines and amines containing substituents, such as naturally occurring and synthetic amines containing substituents. Other pharmaceutically acceptable non-toxic organic bases capable of forming salts include ion exchange resins and arginine, betaine, caffeine, choline, N', N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, reduced glucosamine, glucosamine, histidine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, chloroprocaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and the like.

When compound provided by the invention is alkali, can be from pharmaceutically acceptable nontoxic acid, including inorganic acid and organic acid, conveniently make its corresponding salt.Such acid includes, as, acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, formic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, nitric acid, bashing acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, oxalic acid, propionic acid, glycolic acid, hydroiodic acid, perchloric acid, cyclamic acid, salicylic acid, 2-naphthalenesulfonic acid, saccharinic acid, trifluoroacetic acid, tartaric acid and p-toluenesulfonic acid etc.Preferably, citric acid, hydrobromic acid, formic acid, hydrochloric acid, maleic acid, phosphoric acid, sulfuric acid and tartaric acid.More preferably, formic acid and hydrochloric acid.

Prodrugs of the compounds of the present invention are included within the scope of the present invention. Generally, the prodrug refers to a functional derivative that is easily converted into the desired compound in vivo. For example, any pharmaceutically acceptable salt, ester, salt of an ester or other derivative of the compounds of the present application, which can directly or indirectly provide the compounds of the present application or their pharmaceutically active metabolites or residues after administration to a receptor.

The compounds of the present invention may contain one or more asymmetric centers and may give rise to diastereomers and optical isomers. The present invention includes all possible diastereomers and racemic mixtures thereof, their substantially pure resolved enantiomers, all possible geometric isomers and pharmaceutically acceptable salts thereof.

When the compound represented by formula (I) exists in tautomerism, unless otherwise stated, the present invention includes any possible tautomerism and pharmaceutically acceptable salts thereof, and mixtures thereof.

When the compounds of formula (I) are substituted with heavier isotopes such as deuterium, they may afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements.

The term "pharmaceutical composition" refers to a mixture of one or more compounds of the present application or pharmaceutically acceptable salts thereof and pharmaceutically acceptable excipients. The purpose of a pharmaceutical composition is to facilitate administration of the compounds of the present application to an organism.

In the present invention, "a", "an", "the", "at least one" and "one or more" are used interchangeably. Thus, for example, a mixture comprising "a" pharmaceutically acceptable excipient composition can be interpreted as indicating that the pharmaceutical composition includes "one or more" pharmaceutically acceptable excipients.

The term "pharmaceutically acceptable excipients" refers to those excipients that have no significant irritation to the organism and do not impair the biological activity and performance of the active compound. Suitable excipients are well known to those skilled in the art, such as carbohydrates, waxes, water-soluble and/or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, etc.

The pharmaceutical composition of the present invention can be prepared by combining the compound of the present application with suitable pharmaceutically acceptable excipients, for example, it can be formulated into solid, semi-solid, liquid or gaseous preparations, such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, suppositories, injections, inhalants, gels, microspheres and aerosols.

Typical routes of administration of the compounds of the invention or their pharmaceutically acceptable salts or their pharmaceutical compositions include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, and intravenous administration.

The term "treat" generally refers to obtaining a desired pharmacological and/or physiological effect. The effect may be therapeutic in terms of partial or complete stabilization or cure of a disease and/or side effects resulting from a disease. As used herein, "treat" encompasses any treatment of a patient's disease that: (a) inhibits the symptoms of a disease, i.e., arrests its development; or (b) alleviates the symptoms of a disease, i.e., causes regression of the disease or symptoms.

The term "effective amount" means the amount of the compound of the present application that (i) treats or prevents a specific disease, condition or disorder, (ii) alleviates, ameliorates or eliminates one or more symptoms of a specific disease, condition or disorder, or (iii) prevents or delays the onset of one or more symptoms of a specific disease, condition or disorder described herein. The amount of the compound of the present application that constitutes a "therapeutically effective amount" varies depending on the compound, the disease state and its severity, the mode of administration, and the age of the mammal to be treated, but can be routinely determined by those skilled in the art based on their own knowledge and the present disclosure.

DETAILED DESCRIPTION

In order to make the above content clearer and more specific, the present invention will further illustrate the technical scheme of the present invention with the following examples. The following examples are only used to illustrate the specific embodiments of the present invention so that those skilled in the art can understand the present invention, but are not used to limit the scope of protection of the present invention. In the specific embodiments of the present invention, the technical means or methods not specifically described are conventional technical means or methods in the art.

Unless otherwise specified, all temperatures herein are in degrees Celsius.

The following abbreviations are used in the examples:

DMF: N,N-dimethylformamide;

NIS: N-iodosuccinimide;

THF: tetrahydrofuran;

EA: ethyl acetate;

PE: petroleum ether;

DCM: dichloromethane;

MeOH: methanol;

m-CPBA: meta-chloroperbenzoic acid;

CsF: cesium fluoride;

TBDPSCl: tert-butyldiphenylchlorosilane;

DPPA: diphenylphosphoryl azide;

CDI: N,N'-carbonyldiimidazole;

DIEA: N,N-diisopropylethylamine;

CataCXium A Pd G3: [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate; DBU: 1,8-diazabicyclo[5.4.0]undec-7-ene;

PyBOP: 1H-benzotriazol-1-yloxytripyrrolidino hexafluorophosphate;

LAH: lithium aluminum tetrahydride;

TFA: trifluoroacetic acid;

t-BuOK: potassium tert-butoxide;

t-BuOH: tert-butyl alcohol;

Tol: toluene;

TMP: 2,2,6,6-tetramethylpiperidine;

n-BuLi: n-butyllithium;

Tf ₂ O: trifluoromethanesulfonic anhydride;

SOCl ₂ : thionyl chloride;

CH ₃ CN: acetonitrile;

POCl ₃ : phosphorus oxychloride;

TEA: triethylamine;

NaH: sodium hydride;

Pd(PPh ₃ ) ₄ : tetrakistriphenylphosphine palladium;

HCl: hydrochloric acid;

H ₂ SO ₄ : sulfuric acid;

t-BuONa: sodium tert-butoxide;

NaOMe: sodium methoxide;

EtOH: ethanol;

Grubbs2: 1,3-bis(2,4,6-trimethylphenyl)-2-(imidazolidinylidene)(dichlorobenzylidene)(tricyclohexylphosphine)ruthenium; Togni's reagent II: 1-(trifluoromethyl)-1,2-benzimidoyl-3(1H)-one;

TBAI: tetrabutylammonium iodide;

BINAP: 1,1'-binaphthyl-2,2'-bis(diphenylphosphine);

Pd(dppf)Cl ₂ DCM: 1,1'-bis(diphenylphosphino)ferrocenepalladium(II) dichloride dichloromethane complex;

[Rh(COD)Cl] ₂ : (1,5-cyclooctadiene)rhodium(I) chloride dimer;

K ₃ PO ₄ : potassium phosphate;

KOAc: potassium acetate;

Na ₂ CO ₃ : sodium carbonate;

CuI: cuprous iodide;

DMF-DMA: N,N-dimethylformamide dimethyl acetal;

LiHMDS: lithium bis(trimethylsilyl)amide;

MTBE: methyl tert-butyl ether;

DMAP: 4-dimethylaminopyridine.

Synthesis of intermediate M1:

Step 1: Synthesis of compound M1-1

At room temperature, compound M1-0 (208 g) was dissolved in anhydrous MeOH (2 L), thionyl chloride (286 mL) was added dropwise at 0 ° C, and the temperature was controlled at 5 ° C for 1 hour. After the reaction, the reaction solution was concentrated and diluted with anhydrous DCM (1 L). At 0 ° C, the diluted liquid was added dropwise to a saturated sodium bicarbonate solution, separated, and the organic layer was washed with saturated brine (500 mL), dried over anhydrous sodium sulfate, and concentrated. The concentrate was purified by column chromatography (EA: DCM = 0-50%) to obtain the product M1-1 (240 g, 95% yield). ESI-MS m/z = 258.1 [M + H] ⁺ .

Step 2: Synthesis of Compound M1-2

At room temperature, compound M1-1 (235 g) was dissolved in anhydrous THF (2.4 L), and lithium aluminum tetrahydride (69.4 g) was added in batches at 0°C. After the addition, the mixture was stirred at 60°C for 30 min. After the reaction was completed, the reaction solution was cooled, and water (69.4 mL) was added dropwise under an ice bath, and then a 15% aqueous sodium hydroxide solution (69.4 mL) was added dropwise, and finally water (208.2 mL) was added dropwise, and anhydrous sodium sulfate was added for drying. The filtrate was filtered and concentrated to obtain product M1-2 (165 g, 90% yield), which was directly used in the next step. ESI-MS m/z=202.1[M+H] ⁺ .

Step 3: Synthesis of Compound M1-3

At room temperature, compound M1-2 (160 g) was dissolved in trifluoroacetic acid (500 mL), water (67 mL) was added, and the mixture was reacted at 60°C overnight. The reaction liquid was concentrated to obtain crude product M1-3 (320 g, 259%), which was directly used in the next step. ESI-MS m/z=156.1 [M+H] ⁺ .

Step 4: Synthesis of Compound M1-4

At room temperature, compound M1-3 (308 g) was dissolved in DMF (350 mL), imidazole (540 g) was added at 0°C, and TBDPSCl (170 mL) was added dropwise. After the addition, the mixture was stirred at room temperature for 1 hour. After the reaction was completed, water and EA were added to dilute the mixture, and the aqueous phase was extracted with EA for 3 times. The organic phases were combined, washed with saturated brine for 3 times, dried over anhydrous sodium sulfate, and concentrated. The concentrate was purified by column chromatography (EA:PE=0-15%) to obtain product M1-4 (192 g, 25% yield). ESI-MS m/z=394.1[M+H] ⁺ .

Step 5: Synthesis of Compound M1

At room temperature, compound M1-4 (187 g) and difluoromethyl (2-pyridyl) sulfone (184 g) were dissolved in anhydrous DMF (1.4 L), and a DMF (460 mL) solution of potassium tert-butoxide (107 g) was added dropwise at -50 ° C. After the addition was completed, the temperature was controlled at -40 ° C for 2 hours. After the reaction was completed, a saturated ammonium chloride solution was added dropwise at -50 ° C until the solution became weakly acidic, and the temperature was naturally raised to room temperature for 18 hours. The filtrate was filtered to obtain a filtrate, EA (1.4 L) was added for dilution, and the filtrate was filtered again to obtain a filtrate, which was concentrated. The concentrate was purified by column chromatography (MeOH: DCM = 0-10%) to obtain the product M1 (60 g, 67% yield). ¹ H NMR (500MHz, DMSO-d6) δ3.95-3.92(m,1H),3.70-3.67(m,1H),3.32-3.27(m,2H),2.94-2.89(m,1H),2.69-2.66(m,1H),2.50-3.45(m,1H),1.99-1. 92(m,2H),1.88-1.75(m,2H). ESI-MS m/z=190.1[M+H] ⁺ .

Synthesis of intermediate M2:

Step 1: Synthesis of compound M2-1

At room temperature, 2-chloro-3-fluoro-pyridine-4-carboxylic acid (54.00 g), toluene (390.00 mL), tert-butyl alcohol (390.00 mL), triethylamine (128.27 mL), powdered 4A molecular sieve (90.00 mL) (pre-activated) were added in sequence, and the mixture was kept at reflux for half an hour (internal temperature 87° C.) under nitrogen protection. The mixture was then naturally cooled to room temperature, and then DPPA (99.44 mL) was added, the temperature was raised to reflux, and the mixture was kept at reflux for 5 hours. The reaction mixture was cooled to below 40°C, and then EA 500mL was added for dilution; the mixture was further cooled to room temperature, and the added molecular sieve was filtered out with diatomaceous earth as the filter aid; the residue was rinsed with EA 1500mL for several times and then dried; the filtrate was collected, and washed and separated with water 700mL and saturated brine 700mL in turn; the organic phase was dried over anhydrous sodium sulfate; the mixture was filtered, the desiccant was removed, and the concentrate was purified by column chromatography (PE/EA=30:1-20:1), and the eluent was concentrated to finally obtain the product M2-1 (68.2g, yield 89.88%). ESI-MS m/z: 247.1[M+H] ⁺ .

Step 2: Synthesis of compound M2-2

At room temperature, compound M2-1 (65.00 g) was dissolved in CH ₃ CN (82.00 mL), cooled in a water bath, and hydrochloric acid (4M/dioxane) (38.43 g) was slowly added. The reaction was stirred at room temperature for about 16 hours, and a white solid was precipitated in a suspended state. The reaction mixture was filtered, and the filter cake was rinsed with a small amount of acetonitrile, dried, and the filtrate was discarded. The filter cake was collected and added to a mixture of 700 mL of saturated sodium bicarbonate aqueous solution and 700 mL of ethyl acetate, alkalized, extracted, and separated; the aqueous phase was extracted with 350 mL of ethyl acetate again, and separated; the ethyl acetate phase was combined, and 300 mL of saturated sodium chloride aqueous solution was added for washing and separation; the organic phase was dried over anhydrous sodium sulfate, filtered, the desiccant was removed, and concentrated to obtain the product M2-2 (36.3 g, yield 94.0%). ESI-MS m/z: 147.1 [M+H] ⁺ .

Step 3: Synthesis of Compound M2-3

At room temperature, compound M2-2 (36.00 g) was dissolved in acetonitrile (180.00 mL), NIS (66.32 g) and p-toluenesulfonic acid (2.12 g) were added thereto, and the mixture was heated to 70°C under nitrogen protection for reaction. The reaction solution was cooled to 50°C, 900 mL of water was added, and a white solid powder was precipitated. The mixture was slurried for half an hour; filtered, and the filter cake was rinsed with water and dried. The filter cake was collected, 1200 mL of ethyl acetate was added to dissolve it completely, and then it was washed twice with 350 mL of saturated sodium sulfite aqueous solution, and then washed and separated with 350 mL of saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the product M2-3 (63.2 g, yield 94.43%). ESI-MS m/z: 272.9 [M+H] ⁺ .

Step 4: Synthesis of Compound M2-4

At room temperature, compound M2-3 (57.50 g) was dissolved in DMF (22.00 mL), zinc cyanide (32.22 g), tetrakistriphenylphosphine palladium (12.19 g) and powdered Molecular sieves (20.00 mL) were added, and the mixture was heated and kept at 100 °C in a nitrogen atmosphere for about 7 hours. The oil bath was removed, and the mixture was cooled naturally to room temperature and waited for post-treatment. The reaction mixture was filtered with diatomaceous earth as the filter aid, and then dried; the filtrate was collected and concentrated at 60-70 °C to obtain a pale yellow solid crude product. The filter residue was rinsed and dried with 500 mL of ethyl acetate; the rinse liquid was collected, combined with the crude product, and concentrated again until no liquid was distilled out; 700 mL of ethyl acetate was added to dissolve and concentrate the obtained solid crude product, and then 250 mL of saturated sodium chloride was used each time, washed 3 times, and separated. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a pale yellow solid. 160 mL of PE/EA＝3/1 mixture was added, slurried for half an hour, filtered, and dried. The filter cake was collected, concentrated in a 45 °C water bath, and then pulled to constant weight with a high vacuum oil pump; the product M2-4 (36.1 g, yield 99.7%) was finally obtained. ESI-MS m/z: 172.0 [M+H] ⁺ .

Step 5: Synthesis of Compound M2-5

At room temperature, concentrated sulfuric acid (61.37 mL) was added to a 500 mL single-mouth flask, cooled to below 10 ° C in an ice-water bath, and compound M2-4 (39.30 g) was added in batches. After addition, it was stirred for 10 minutes, and kept warm at 60 ° C in an oil bath in a nitrogen atmosphere, and reacted for about 1 hour. The reaction solution was cooled to room temperature, and then carefully added to 1100 mL of ice-water mixture, diluted and quenched, and a small amount of yellow solid precipitated. After stirring for 10 minutes, filter; collect the filter cake, use 50 mL of saturated sodium bicarbonate aqueous solution, beat for 20 minutes, filter again, collect the two filtrates, and combine; then slowly add sodium carbonate solid, adjust the pH to about 7, and a white solid powder precipitates. Stir for half an hour, filter, and drain; rinse the filter cake with 100 mL of water each time, drain, and rinse twice. Collect the filter cake, put it in a vacuum oven, and dry it at 55 ° C to constant weight to obtain the product M2-5 (33.6 g, yield 77.37%). ESI-MS m/z: 190.0 [M+H] ⁺ .

Step 6: Synthesis of Compound M2-6

At room temperature, tetrahydrofuran (470.00 mL) was added, and after nitrogen replacement, sodium hydride (10.00 g) was added under the protection of a slight nitrogen flow, and heated in an oil bath, kept at 40-45°C, and stirred for 15 minutes; then compound M2-5 (18.95 g) was added in batches, and after addition, mechanical stirring was kept for 20 minutes, and then CDI (24.31 g) was carefully added in batches, and after addition, stirring was completed for 15 minutes, the oil bath was heated and heated, and the reaction was kept under reflux. The reaction solution was cooled to below 10°C with an ice water bath, and then 500 mL of saturated ammonium chloride aqueous solution was added. A light yellow solid precipitated, and 1000 mL of water was added; then it was transferred to a 5L beaker, and 3000 mL of water was added; it was stirred for 1 hour, filtered, and drained; the filter cake was collected, placed in a vacuum oven, and dried at 50-55°C to constant weight to obtain the product M2-6 (18.3 g, yield 84.93%). ESI-MS m/z: 216.0 [M+H] ⁺ .

Step 7: Synthesis of Compound M2

At room temperature, compound M2-6 (18.00 g) and DIEA (36.00 mL) were dissolved in POCl ₃ (180.00 mL), and heated to 100°C for about 2.5 hours under nitrogen atmosphere. The phosphorus oxychloride was removed by concentration under reduced pressure, and DCM 100 mL was used twice; the concentrated residue was dissolved with 400 mL of dichloromethane, and then added dropwise to 500 mL of saturated sodium bicarbonate aqueous solution, and cooled with ice water; after stirring for 15 minutes, the liquid was separated; the aqueous phase was extracted with 300 mL of dichloromethane and separated; the dichloromethane phases were combined, washed with 300 mL of saturated sodium chloride aqueous solution, and separated; dried over anhydrous sodium sulfate, filtered, and concentrated. The concentrate was purified by silica gel column (PE/EA=90/10-75/25) to obtain product M2 (10.95 g, yield 51.94%). ESI-MS m/z: 251.9 [M+H] ⁺ .

Synthesis of intermediate M3:

Step 1: Synthesis of compound M3-1

At room temperature, compound M3-0 (500 mg), styrene (373 mg) and Grubbs 2nd generation catalyst (405.72 mg) were dissolved in anhydrous DCM (20 mL) and reacted overnight at 50° C. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the concentrate was separated and purified by column chromatography to obtain the target compound M3-1 (400 mg).

Step 2: Synthesis of Compound M3

At room temperature, compound M3-1 (400 mg) was dissolved in anhydrous THF (10 mL), LAH (160 mg) was added at 0°C, and the temperature was raised to 70°C for reaction for 30 min. After the reaction was completed, the temperature was lowered to 0°C, 170 uL of water was slowly added, then 170 uL of 15% sodium hydroxide aqueous solution was added, and finally 510 uL of water was added, and the reaction was carried out at room temperature for 15 min, anhydrous sodium sulfate was added for drying, diatomaceous earth was filtered, and the filtrate was concentrated to obtain the target compound M3 (277 mg).

Synthesis of intermediate M4:

Step 1: Synthesis of compound M4-1

At room temperature, compound M3-0 (800 mg), p-chlorostyrene (795 mg) and Grubbs 2nd generation catalyst (325 mg) were dissolved in anhydrous DCM (20 mL) and reacted at 50°C overnight. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the concentrate was separated and purified by column chromatography to obtain the target compound M4-1 (729 mg). ESI-MS m/z = 320.1 [M+H] ⁺ .

Step 2: Synthesis of Compound M4

At room temperature, compound M4-1 (729 mg) was dissolved in anhydrous THF (10 mL), LAH (260 mg) was added at 0°C, and the temperature was raised to 70°C for reaction for 30 min. After the reaction was completed, the temperature was lowered to 0°C, 280 uL of water was slowly added, then 280 uL of 15% sodium hydroxide aqueous solution was added, and finally 840 uL of water was added, and the reaction was carried out at room temperature for 15 min, anhydrous sodium sulfate was added for drying, diatomaceous earth was filtered, and the filtrate was concentrated to obtain the target compound M4 (563 mg). ESI-MS m/z=264.1[M+H] ⁺ .

Synthesis of intermediate M5:

Step 1: Synthesis of compound M5-1

At room temperature, compound M3-0 (400 mg), methylenecyclopentane (236 mg) and Grubbs 2nd generation catalyst (325 mg) were dissolved in anhydrous DCM (20 mL) and reacted overnight at 50° C. After the reaction was completed, the reaction solution was concentrated to a small amount under reduced pressure and directly wet loaded for column chromatography purification to obtain the target compound M5-1 (410 mg).

Step 2: Synthesis of Compound M5

Compound M5-1 (410 mg) was dissolved in anhydrous THF (10 mL), LAH (204 mg) was added at 0°C, and the temperature was raised to 70°C for reaction for 30 min. After the reaction was completed, the temperature was lowered to 0°C, 210 uL of water was slowly added, 210 uL of 15% sodium hydroxide aqueous solution was added, and 630 uL of water was added, and the reaction was carried out at room temperature for 15 min. Anhydrous sodium sulfate was added for drying, diatomaceous earth was filtered, and the filtrate was concentrated to obtain the target compound M5 (370 mg).

Synthesis of intermediate M6:

Step 1: Synthesis of compound M6-1

At room temperature, M3-0 (1.00 g) and 2,3-dimethyl-2-ene (2.01 g) were added to DCM (20.00 mL), and Grubbs 2nd generation catalyst (0.41 g) was added. _N2 was replaced three times, and under _N2 protection, the reaction was heated to 50°C and stirred for 20 hours. The reaction was cooled and concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain the target compound M6-1 (0.2 g, yield 9.04%).

Step 2: Synthesis of Compound M6

At room temperature, M6-1 (0.20 g) was added to THF (4.00 mL), and LAH (0.10 g) was added in batches under ice bath. The reaction temperature was raised to 70°C and stirred for 1 hour. The reaction was cooled, and H ₂ O (100 uL) was added dropwise under ice bath, followed by 15% NaOH (100 uL) solution, and finally H ₂ O (300 uL) was added dropwise, and stirred at room temperature for 10 minutes, and then anhydrous sodium sulfate was added to dry and stirred for 5 minutes. Filter, rinse the filter cake with EA, and concentrate the filtrate under reduced pressure to obtain the target compound M6 (0.18 g). Synthesis of intermediate M7:

Step 1: Synthesis of compound M7-1

At room temperature, compound M3-0 (0.5 g), Togni's reagent II (1.51 g) and tetrabutylammonium iodide (0.44 g) were dissolved in 1,4-dioxane (10 mL), nitrogen was replaced, and the reaction was carried out at 80 degrees for 10 hours. The reaction solution was directly concentrated, and the concentrate was separated and purified by chromatography column (DCM/EA=2/1) to obtain product M7-1 (0.45 g, 68% yield).

Step 2: Synthesis of Compound M7

At room temperature, compound M7-1 (0.3 g) was dissolved in tetrahydrofuran, and lithium aluminum tetrahydride (123 mg) was added under an ice bath, followed by a tetrahydrofuran solution of borane (3.2 mL). After reacting for 1 minute, the reaction was immediately quenched with water until no more bubbles were generated, 15% sodium hydroxide (0.3 mL) was added, and then water (0.9 mL) was added. After stirring and quenching was complete, sodium sulfate was added for drying, the filtrate was filtered, and concentrated to obtain the product M7 (80 mg, 33% yield).

Synthesis of intermediate M8:

At room temperature, compound M7-1 (0.3 g) was dissolved in tetrahydrofuran, and lithium aluminum tetrahydride (123 mg) was added under ice bath, and then borane tetrahydrofuran solution (3.2 mL) was added, and the ice bath was removed to react for 5 minutes. The reaction was complete after LC-MS monitoring, and the reaction was quenched with water until no more bubbles emerged, 15% sodium hydroxide (0.3 mL) was added, and then water (0.9 mL) was added. After stirring and quenching was complete, sodium sulfate was added to dry, and the filtrate was filtered and dried to obtain product M8 (164 mg, 73% yield). Synthesis of intermediate M9:

Step 1: Synthesis of compound M9-1

At room temperature, di(imidazole-1-yl)methanone (2.70 g) was added to THF (20 mL) of compound 2-amino-4-bromo-5-chloro-3-fluorobenzoic acid (4.0 g), and then N-ethyl-N-isopropylpropan-2-amine (1.4 g) was added thereto, and the mixture was moved to 50°C for reaction for 2 hours. Compound 2-amino-4-bromo-5-chloro-3-fluorobenzoic acid was basically completely converted into an intermediate product, and then the mixture was added dropwise to ice ammonia water (35 mL), and the reaction was complete after stirring for 5 minutes. The mixture was poured into ice water, and the mixture was extracted with ethyl acetate. The organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo. The residue was purified by flash silica gel column chromatography (petroleum ether/ethyl acetate=70:30) to obtain the desired target product compound M9-1 (1.6 g) as a brown solid.

Step 2: Synthesis of compound M9-2

At room temperature, the intermediate M9-1 (19.00 g) was added to tetrahydrofuran (200.00 mL), and sodium hydride (7.10 g) was added in batches at room temperature. Under _N2 protection, the mixture was heated to 40°C and stirred for 0.5 hours. Di(1H-imidazol-1-yl)phosphine (18.99 g) was added in batches. The mixture was heated to 70°C and stirred for 0.5 hours. The reaction was completed by LCMS monitoring. Saturated _NH4Cl (20 mL) was added to quench the reaction, and then the pH was adjusted to 5-6 with 1N HCl, and then extracted twice with EA (80 mL). The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate, and concentrated to obtain 17.4 g of yellow powder crude product M9-2, which was used directly in the next step.

Step 3: Synthesis of Compound M9

At room temperature, M9-2 (16.40 g) was added to methanol (300.00 mL) and then sodium methoxide (4.29 g), and finally iodomethane (4.95 mL) was added dropwise, and stirred at room temperature for 0.5 hours. H ₂ O (20 mL) was added to dilute, and then the pH was adjusted to 5-6 with 2N HCl, and concentrated under reduced pressure. The crude product was added to H ₂ O (20 mL) and stirred for 5 minutes, filtered, and the filter cake was washed with water and then washed with PE/EA=3/1 (20 mL) for 3 times, and dried to obtain a yellow powder product M9 (14.6 g). ESI-MS m/z: 323 [M+H] ⁺ .

Synthesis of intermediate M10:

Step 1: Synthesis of compound M10-1

4-Bromo-2-fluoroaniline (11.0 g), potassium carbonate (20.18 g), and KI (9.7 g) were added to NMP (110.0 mL) respectively, and 1-(chloromethyl)-4-methoxybenzene (16.24 mL) was added dropwise, and stirred at room temperature for 7 hours. The reaction solution was poured into H ₂ O (120 mL), and then extracted twice with MTBE (60 ml) (three layers, and the upper layer was taken), and the organic layer was washed twice with saturated brine and dried over anhydrous sodium sulfate. The crude product was slurried with PE (60 ml) for 1 hour. Filter, rinse the filter cake with PE, and dry to obtain compound M10-1 (17.68 g, 70.33% yield). ESI-MS m/z＝430[M+H] ⁺ .

Step 2: Synthesis of compound M10-2

Add the above compound M10-1 (6.3 g) to the reaction flask, then add bis(pinacolato)diboron (11.2 g), potassium acetate (2.88 g) and Pd(dppf)Cl ₂ .DCM (1.20 g), and react at 100°C for 12 h. TLC confirms that the raw material is completely converted. Add 30 mL of ethyl acetate, then wash and separate with 20 mL of saturated sodium chloride aqueous solution × 2. Add anhydrous sodium sulfate to dry. Filter and concentrate, and the concentrate is separated and purified by column chromatography (PE～PE:EA＝10:1) to obtain compound M10-2 (4.14 g, 67.5% yield). ESI-MSm/z＝478[M+H] ⁺ .

Step 3: Synthesis of compound M10-3

Add the product M10-2 (4.1 g) from the previous step to the reaction flask, add dioxane (50 mL) and water (10 mL) to dissolve, then add 4-methylcyclo-2-ene-1-one (3.2 g), BINAP (1.22 g), [Rh(COD)Cl] ₂ (483 mg), K ₃ PO ₄ (6.2 g), replace with nitrogen, and react at 35°C for 10 min. TLC monitoring shows that the raw material has reacted completely, add ethyl acetate 40 mL, and then wash and separate with saturated sodium chloride aqueous solution 30 mL × 2. Add anhydrous sodium sulfate to dry. Filter and concentrate, and the concentrate is separated and purified by column chromatography (PE～PE:EA＝10:1) to obtain compound M10-3 (2.84 g, 62.8% yield). ESI-MS m/z＝462[M+H] ⁺ .

Step 4: Synthesis of compound M10-4

The product M10-3 (2.6 g) from the previous step was added to the reaction flask, acetonitrile (25 mL) was added to dissolve, NIS (2.05 g) and trifluoroacetic acid (68 mg) were added, and the mixture was reacted at room temperature for 2 h. The solvent was concentrated, and the concentrate was separated and purified by column chromatography (PE-PE:EA=8:1) to obtain compound M10-4 (1.22 g, 33.7% yield). ESI-MS m/z=588[M+H] ⁺ .

Step 5: Synthesis of compound M10-5

Add M10-4 (1.2 g) to the reaction flask, add DMF (12 mL) to dissolve, then add methyl 2,2-difluoro-2-fluorosulfonyl acetate (1.56 g) and cuprous iodide (1.16 g), and react at 85°C for 2 h under nitrogen protection. Add 30 mL of ethyl acetate, then wash and separate with 25 mL × 3 saturated sodium chloride aqueous solution. Add anhydrous sodium sulfate to dry. Filter and concentrate, and the concentrate is separated and purified by column chromatography (PE~PE:EA＝3:1) to obtain compound M10-5 (600 mg, 55.6% yield). ESI-MS m/z＝530[M+H] ⁺ .

Step 6: Synthesis of Compound M10-6

Compound M10-5 (600 mg) was added to the reaction flask, THF (10 mL) was added to dissolve, and the mixture was cooled to -78°C under nitrogen protection. Then LiHMDS (2.3 mL) was slowly added, and the mixture was kept warm for 0.4 h after the addition. Then ethyl cyanoformate (246 mg) was added at the same temperature, and the mixture was reacted for 0.5 h after the addition. TLC monitoring showed that the raw material reacted completely, and 5 mL of ammonium chloride aqueous solution was added at low temperature to quench the reaction. Then 20 mL of ethyl acetate was added, and the mixture was washed and separated with 20 mL of saturated sodium chloride aqueous solution × 2. Anhydrous sodium sulfate was added to dry the mixture. Filtered, and the filtrate was concentrated to obtain the crude product M10-6 (600 mg 88.3% yield). ESI-MS m/z=602[M+H] ⁺ .

Step 7: Synthesis of Compound M10-7

The above compound M10-6 (600 mg) was added to ethanol (10 mL) and water (2 mL), and sodium bicarbonate (2.0 g) and methyl isothiourea sulfate (2.2 g) were added. The reaction solution was stirred at 50°C for 4 hours. The reaction solution was added to 25 mL of water and extracted twice with ethyl acetate (15 mL). The combined organic phase was washed twice with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The concentrate was separated and purified by silica gel column chromatography (PE: EA = 100: 0-1: 1) to obtain compound M10-7 (480 mg). ESI-MS m/z = 628 [M + H] ⁺ .

Step 8: Synthesis of Compound M10

Compound M10-7 (480 mg) was dissolved in dichloromethane (5 mL), N, N-diisopropylethylamine (0.3 g) was added, the temperature was lowered to 0-10 ° C, trifluoromethanesulfonic anhydride (0.3 g) was slowly added to the reaction solution, and the reaction was carried out at this temperature for 15 minutes. The reaction solution was poured into a saturated aqueous ammonium chloride solution (5 mL), separated, the aqueous phase was extracted twice with dichloromethane (5 mL), the combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The concentrate was slurried with a mixed solvent (PE: MTBE = 20: 1, 4 mL), filtered, and the filter cake was dried to obtain compound M10. ESI-MS m/z = 760 [M + H] ⁺ .

Synthesis of intermediate M11:

Step 1: Synthesis of compound M11-1

At room temperature, the compound 2,5-dihydro-1H-pyrrole-1-carboxylic acid tert-butyl ester (20g) was dissolved in a mixed solution of tert-butyl alcohol (200mL) and water (200mL), and potassium osmate dihydrate (1.74g) and N-methylmorpholine oxide (51.5g) were added in sequence, and stirred at 45°C for 15 hours. The reaction solution was directly concentrated, extracted with ethyl acetate (50mL*2), and washed with saturated sodium sulfite solution (60mL). The combined organic layer was washed with saturated sodium chloride (50mL*3), dried over anhydrous sodium sulfate, filtered, and concentrated. The concentrate was separated and purified by column chromatography (petroleum ether/ethyl acetate = 1/0 to 0/1) to obtain compound M11-1 (19.0g). ¹ H NMR (400MHz, CDCl ₃ ) δ 4.20 (m, 2H), 3.57-3.54 (m, 2H), 3.32-3.29 (m, 2H), 2.85-2.80 (m, 2H), 1.40 (s, 9H).

Step 2: Synthesis of compound M11-2

At room temperature, compound M11-1 (19 g) was dissolved in dichloromethane (150 mL), cooled to 0°C, and then iodobenzene diacetate (20.0 g) was added, and the reaction system was transferred to room temperature and stirred for 3 hours. Saturated sodium bicarbonate solution (40 mL) was added to quench the reaction system, and dichloromethane (10 mL) was added and stirred for 0.5 hours. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. Methyl tert-butyl ether (20 mL) was added at 25°C, and stirred for 10 minutes, filtered, and concentrated under reduced pressure to obtain compound M11-2 (19.5 g).

Step 3: Synthesis of compound M11-3

At room temperature, compound M11-2 (19.5 g) was dissolved in tetrahydrofuran (50 mL), cooled to -78 ° C, and ethylmagnesium bromide (1 M, 150 mL) was added to the reaction system. The reaction system was warmed to room temperature and stirred for 15 hours. The reaction system was then cooled to 10 ° C, saturated ammonium chloride solution (100 mL) was added thereto to quench the reaction system, and extracted with ethyl acetate. The organic phase was washed three times with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The concentrate was separated and purified by column chromatography to obtain compound M11-3 (18.9 g).

Step 4: Synthesis of compound M11-4

At room temperature, compound M11-3 (18.9 g) was dissolved in dichloromethane (200 mL), the reaction system was moved to 0 ° C and DBU (4.5 g) and 2,2,2-trichloroacetonitrile (53 g) were added, the reaction system was transferred to room temperature and stirred for 16 hours. The residue was concentrated under reduced pressure, extracted twice with ethyl acetate, washed twice with water and saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The concentrate was separated and purified by column chromatography (petroleum ether/ethyl acetate = 1/0 to 5/1) to obtain compound M11-4 (17.5 g).

Step 5: Synthesis of compound M11-5

At room temperature, compound 2-phenylpropane-2-amine (6.5 g) was dissolved in DCE (140 mL), and 1,5-cyclooctadiene iridium chloride dimer (2.5 g) was added thereto. The reaction system was cooled to 0°C, compound M11-4 (17.5 g) was dissolved in DCE (200 mL) and added to the above reaction system, the reaction was heated to room temperature and continued to stir for 15 hours, and the reaction solution was directly concentrated. The concentrate was separated and purified by column chromatography (petroleum ether/ethyl acetate = 1/0 to 10/1) to obtain compound M11-5 (15.1 g). ESI-MS m/z = 357 [M+H] ⁺ .

Step 6: Synthesis of Compound M11-6

At room temperature, M11-5 (15.1 g) was dissolved in toluene (380 mL), and then the second-generation Grubbs catalyst (1.2 g) was added. The reaction system was heated to 125° C. and stirred for 16 hours. The reaction solution was directly filtered, and the filtrate was concentrated to obtain a crude product. The crude product was separated and purified by column chromatography (petroleum ether/ethyl acetate=1/0 to 10/1) to obtain compound M11-6 (12.1 g). ESI-MS m/z=329[M+H] ⁺ .

Step 7: Synthesis of Compound M11-7

At room temperature, compound M11-6 (12.1 g) was dissolved in methanol (90 mL), followed by addition of HCl/MeOH (4 M, 35 mL). The reaction system was heated to 35°C and stirred for 16 hours. The pH value of the reaction mixture was adjusted to 12, and extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain M11-7 (11.0 g). ESI-MS m/z=229[M+H] ⁺ .

Step 8: Synthesis of compound M11-8

At room temperature, compound M11-7 (11 g) was dissolved in THF (100 mL), followed by the addition of 9-fluorenylmethyl chloroformate (9.0 g) and sodium carbonate (10.0 g). The mixture was stirred at 0°C for 1 hour. It was extracted twice with ethyl acetate and washed once with water. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give compound M11-8 (9.8 g). ESI-MS m/z=451[M+H] ⁺ .

Step 9: Synthesis of compound M11-9

At room temperature, compound M11-8 (9.8 g) was dissolved in TFA (180 mL), heated to 75 ° C and stirred for 16 hours. Water (20.0 mL) was added, and the pH was adjusted to 9, and then dichloromethane (20.0 mL) was added for extraction, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product. The crude product was slurried with n-heptane (6 mL) at 25 ° C to obtain M11-9 (7.4 g).

Step 10: Synthesis of Compound M11-10

At room temperature, compound M11-9 (7.4 g) was dissolved in tetrahydrofuran (70 mL), followed by the addition of di-tert-butyl dicarbonate (5.0 g) and triethylamine (6.0 g) and stirred at 25 ° C for 1 hour. Ethyl acetate and water were added for extraction. The organic phases were combined, washed with saturated brine (50.0 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The concentrate was purified by column chromatography (petroleum ether/ethyl acetate = 1/0 to 3/1) to obtain intermediate M11-10 (6.9 g). ESI-MS m/z = 433 [M+H] ⁺ .

Step 11: Synthesis of Compound M11

At room temperature, compound M11-10 (6.9 g) was dissolved in ethanol (70.0 mL), followed by the addition of dimethylamine (36.6 g). The reaction system was stirred at 25 ° C for 3 hours. Directly concentrated under reduced pressure, the concentrate was extracted with ethyl acetate (50.0 mL) and 10% citric acid (200.0 mL), the pH value of the aqueous phase was adjusted to 9, filtered and extracted with ethyl acetate (40.0 mL * 2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain compound M11 (1.6 g, 96.3% purity, 47.7% yield). ESI-MS m / z = 211 [M + H] ⁺ . ¹ H NMR (400MHz, CDCl ₃ ) δ 6.20 (m, 2H), 4.37 (m, 2H), 2.88-3.00 (m, 2H), 2.38 (m, 2H), 1.49 (s, 9H).

Synthesis of intermediate M12:

At room temperature, ethyl 2-methylene-5-oxo-1,3,6,7-tetrahydropyrrolizine-8-carboxylate (10.00 g) was dissolved in THF (150.00 mL) in a reaction bottle, and LAH (2.5 M/THF) (3.63 g) was slowly added, and the temperature was controlled below 60°C. After adding, the mixture was stirred for 0.2 h. The mixture was cooled to 0°C, 3.6 ml of water was added to quench the reaction, and 3.6 ml of 15% aqueous sodium hydroxide solution was added, and finally 10.8 ml of water was added. After stirring for 10 min, anhydrous magnesium sulfate was added to dry the mixture, and the mixture was filtered after stirring for 10 min. The filter cake was washed three times with EA, and the mother liquor was concentrated to obtain the target intermediate M12 (6.5 g, 89.34% yield).

Synthesis of intermediate M13:

The compound ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborol-2-yl)naphthalene-1-yl)ethynyl)triisopropylsilane (10 g) was added to a 100 mL single-mouth bottle, and 30 mL of hydrochloric acid (4M/Dioxane) was added. After reacting at room temperature for half an hour, a saturated sodium bicarbonate solution was added under an ice bath to neutralize the reaction solution, and the filter cake was washed with water twice, dissolved with EA, dried over anhydrous sodium sulfate, and concentrated. The concentrate was purified by column chromatography (PE:EA=82:18) to obtain the target intermediate M13 (9.1 g).

Synthesis of intermediate M14:

Step 1: Synthesis of compound M14-1

Compound M11 (600 mg) was dissolved in dichloromethane (6 mL), the temperature was lowered to 0°C, N,N-diisopropylethylamine (1.41 mL) and M2 (576.29 mg) were added, and the mixture was reacted at room temperature for 0.5 h. After the reaction was complete, the mixture was directly mixed with silica gel and purified by column chromatography to obtain compound M14-1 (785 mg, yield 64.54%). ESI-MS m/z: 426.27 [M+H] +. Step 2: Synthesis of compound M14-2

Compound M12 (251.6 mg) was dissolved in tetrahydrofuran (7 mL), sodium hydride (262.74 mg) was added, the temperature was lowered to 0°C, and a solution of compound M14-1 in tetrahydrofuran (10.00 mL) was slowly added, and the mixture was reacted at room temperature for 3 h. After the reaction was complete, the reaction solution was quenched with saturated ammonium chloride, extracted twice with EA, and purified by column chromatography to obtain compound M14-2 (480 mg, yield 53.83%). ESI-MS m/z: 543.43 [M+H] +.

Step 3: Synthesis of Compound M14

Compound M14-2 (430 mg), intermediate M13 (556.42 mg), methanesulfonic acid [n-butyldi(1-adamantyl)phosphine] (2-amino-1,1'-biphenyl-2-yl) palladium (II) (57.68 mg), potassium phosphate (504.22 mg) were dissolved in 1,4-dioxane (8 mL) and water (2 mL), replaced with nitrogen, and reacted at 100 ° C for 5 h. After the reaction was complete, EA was used for extraction twice and purified by column chromatography to obtain compound M14 (570 mg yield 87.78%). ESI-MS m/z: 849.63 [M+H] +.

Example 1: Synthesis of Compound 3-(4-((1R,5S)-3,8-diazabicyclo[3.2.1]-6-ene-3-yl)-2-((2-(difluoromethylene)tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-pyrano[4,3-d]pyrimidin-7-yl)-4-fluoromethylaniline trifluoroacetate

Step 1: Synthesis of compound 1-1

TMP (20.21 mL) was added to THF (170.00 mL), and _N2 was replaced three times at -10°C. n-BuLi (2.5M/dioxane) (47.91 mL) was added dropwise under _N2 protection, and stirred at -10°C for 10 minutes. The reaction temperature was lowered to -60°C, and a solution of M10-1 (17.18 g) in THF (40.00 mL) was added dropwise, and stirred for 0.5 hours. DMF (30.90 mL) was then quickly added and stirred for 10 minutes. Saturated _NH4Cl (200 ml) was poured into the reaction, and then extracted twice with EA (50 mL). The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. Concentrated under reduced pressure, the crude product was stirred overnight with PE/MTBE = 5/1 (60 ml) for slurrying. After filtration, the filter cake was rinsed with PE/MTBE=5/1 and dried to obtain compound 1-1 (16.59 g, yield 90.66%). ¹ H NMR (500 MHz, CDCl ₃ ) δ10.34 (d, J=0.8 Hz, 1H), 7.19 (dd, J=8.7, 1.4 Hz, 1H), 7.17-7.13 (m, 4H), 6.86-6.80 (m, 5H), 4.23 (s, 4H), 3.78 (s, 6H). ESI-MS m/z=458 [M+H] ⁺ .

Step 2: Synthesis of compound 1-2

Compound 1-1 (16.09 g) and CuI (1.34 g) were added to DMF (80.00 mL), replaced with N ₂ three times, and the reaction temperature was raised to 80 ° C under N ₂ protection. Methyl fluorosulfonyl difluoroacetate (14.97 mL) was added dropwise, and the temperature was raised to 100 ° C and stirred for 1.5 hours. Filter (diatomaceous earth), rinse the filter cake with MTBE (50 ml) three times, wash the filtrate with water, and wash with saturated brine twice. The emulsified layer was extracted with EA (40 mL), and the organic layer was washed with saturated brine three times. Purification by column chromatography (PE/EA, 10-15% EA) gave compound 1-2 (13.11 g, yield 83.46%). ¹ H NMR (500MHz, CDCl ₃ ) δ10.43(q,J＝1.8Hz,1H),7.32(d,J＝8.6Hz,1H),7.17-7.13(m,4H),6.95(t,J＝8.5Hz,1H),6.87-6.82(m,4H),4.38(s,4H),3.79(s,6 H). ESI-MS m/z=448[M+H] ⁺ .

Step 3: Synthesis of Compound 1-3

NaH (1.12 g) was added to THF (50.00 mL) in batches, the temperature was lowered to 0°C under _N2 protection, methyl acetoacetate (3.01 mL) was added dropwise, and the mixture was stirred for 10 minutes. Then n-BuLi (2.5 M/dioxane) (11.18 mL) was added dropwise and stirred for 10 minutes. The mixture was cooled to -15°C and a solution of compound 1-2 (5.00 g) in THF (5.00 mL) was added dropwise, and stirred for 0.5 hours. The reaction mixture was poured into a saturated NH ₄ Cl (100 ml) solution. The mixture was extracted twice with EA (40 ml), and the organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. Compound 1-3 (4.95 g, yield 78.60%) was obtained by column chromatography purification (PE/EA, 30% EA). ESI-MS m/z＝564[M+H] ⁺ .

Step 4: Synthesis of Compound 1-4

Compound 1-3 (4.85 g) was added to DCM (50.00 mL) and then N,N-dimethylformamide dimethyl acetal (2.29 mL). Under N ₂ protection, the mixture was stirred at room temperature for 12 hours, and the intermediate state was generated by LCMS. The mixture was cooled to 0°C and then boron trifluoride etherate (1.95 mL) was added dropwise and stirred for 75 minutes. Saturated NH ₄ Cl solution (80 ml) was poured into the mixture, and then extracted twice with DCM (30 ml). The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. Compound 1-4 (3.95 g, yield 80.02%) was obtained by column chromatography purification (PE/EA, 35% EA). ESI-MS m/z＝574[M+H] ⁺ .

Step 5: Synthesis of Compound 1-5

Compound 1-4 (3.75 g) was added to THF (40.00 mL), and under N ₂ protection, lithium tri-sec-butyl borohydride (7.85 mL, 1.00 mol/L) was added dropwise at -65°C, and stirring was continued for 0.5 hours. The reaction solution was poured into saturated NH ₄ Cl (80 ml), and then extracted twice with EA (30 mL). The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. Purification by column chromatography (PE/EA, 20% EA) gave compound 1-5 (2.11 g, yield 56.07%). ESI-MS m/z＝576[M+H] ⁺ .

Step 6: Synthesis of Compound 1-6

Compound 1-5 (2.01 g) was added to EtOH (40.00 mL) and H ₂ O (4.00 mL), and then 2-methyl-2-mercaptosulfuric acid urea (3.29 g) and Na ₂ CO ₃ (1.30 g). The reaction mixture was heated to 50°C and stirred for 10 hours. The reaction solution was concentrated under reduced pressure, and then EA (30 ml) and H ₂ O (20 mL) were added for extraction twice. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. Purification by column chromatography (PE/EA, 45-50% EA) gave compound 1-6 (794 mg, yield 36.93%). ESI-MS m/z=616[M+H] ⁺ .

Step 7: Synthesis of Compound 1-7

Compound 1-6 (744.00 mg) was added to DCM (10.00 mL), and then DIEA (1.00 mL) was added dropwise. Tf ₂ O (0.61 mL) was added under ice bath. Stir at 0°C for 0.5 hours. The reaction solution was poured into saturated NH ₄ Cl (15 ml), and then extracted twice with DCM (15 ml). The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 1-7 (900 mg). ESI-MS m/z=748[M+H] ⁺ .

Step 8: Synthesis of Compound 1-8

Compound 1-7 (900.00 mg) and M11 (604 mg) were added to DMF (10.00 mL) and then DIEA (0.99 mL). The mixture was heated to 50°C and stirred for 0.5 hours. The mixture was extracted twice with EA (30 ml) and H ₂ O (30 mL). The organic layer was washed three times with saturated brine and dried over anhydrous sodium sulfate. Compound 1-8 (520 mg, yield 55.8%) was obtained by column chromatography purification (PE/EA, 25% EA). ESI-MS m/z=808[M+H] ⁺ .

Step 9: Synthesis of Compound 1-9

Compound 1-8 (500.00 mg) was added to DCM/dichloromethane (7.00 mL) and then m-CPBA (170 mg, 85%) was added in batches and stirred at room temperature for 1.5 hours. The reaction was completed by LCMS monitoring. DCM (20 mL) and saturated NaHCO ₃ solution were added to the reaction and washed twice. The organic layer was then washed with saturated brine and finally dried over anhydrous sodium sulfate. Concentration gave 480 mg, i.e., compound 1-9. The crude product was used directly in the next step. ESI-MS m/z=824[M+H] ⁺ .

Step 10: Synthesis of Compound 1-10

Intermediate M1 (120 mg) was added to THF (3.00 mL) and then t-BuONa (70 mg) was added, and the mixture was stirred at room temperature for 0.5 hours. A solution of compound 1-9 (300 mg) in THF (1.00 mL) was added. The mixture was stirred at room temperature for 0.5 hours. The reaction was monitored by LCMS to be complete. The reaction was quenched by adding saturated NH ₄ Cl (20 mL), and then EA (25 ml) was added and extracted twice. The organic layer was washed with saturated brine and then dried over anhydrous sodium sulfate. Compound 1-10 (150 mg, yield 40.07%) was obtained by column chromatography purification (DCM/MeOH, 5-10% MeOH). ESI-MS m/z＝949[M+H] ⁺ .

Step 11: Synthesis of Compound 1

Compound 1-10 (150 mg) was added to DCM (3.00 mL) and then TFA (3.00 mL) and stirred at room temperature for 0.5 hours. The mixture was concentrated under reduced pressure and the crude product was prepared by HPLC to obtain the trifluoroacetate salt of compound 1 (53 mg, 55.1% yield, 95.05% purity). ¹ H NMR (500 MHz, DMSO-d ₆ )δ10.71(s,1H),9.30(d,J＝10.2Hz,1H),9.21(d,J＝10.0Hz,1H),7.27(d,J＝8.6Hz,1H),6.84(t,J＝8.5Hz,1H),5.03(dd,J＝11.2,4.4Hz,1H),4.84(d,J＝13. 8Hz,1H),4.78-4.60(m,2H),4.67(d,J＝13.8Hz,1H),4.48-4.4 2(m,3H),4.18(d,J＝14.2Hz,2H),4.13-4.07(m,3H),3.91(d,J＝14.6Hz,1H),3.70-3.52(m,3H),3.26-3.12(m,3H),2.86(td,J＝13.1,10.8,7.3Hz,2H ), 2.73 (d, J = 16.1Hz, 1H), 2.24-1.88 (m, 7H), 1.82 (q, J = 11.2Hz, 1H). ESI-MS m/z=609[M+H] ⁺ .

Example 2: Synthesis of Compound 3-(4-((1R, 5S)-3,8-diazabicyclo[3.2.1]oct-6-ene-3-yl)-2-((2-methylenetetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-pyrano[4,3-d]pyrimidin-7-yl)-4-fluoromethylaniline

Step 1: Synthesis of compound 2-1

Intermediate M12 (100 mg) was dissolved in THF (2.50 mL), and t-BuONa (60 mg) was added thereto, and stirred at room temperature for 0.5 hours. A solution of compound 1-9 (270 mg) in THF (1.00 mL) was then added. Stir at room temperature for 0.5 hours. Saturated NH ₄ Cl (20 mL) was added to the reaction solution to quench, and EA (25 ml) was added to extract twice. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate, filtered, and concentrated. The concentrate was purified by column chromatography (DCM/MeOH, 5-10% MeOH) to obtain compound 2-1 (90 mg, yield 28.07%). ESI-MS m/z＝913[M+H] ⁺ .

Step 2: Synthesis of compound 2

Compound 2-1 (90 mg) was dissolved in DCM (2.00 mL), and TFA (2.00 mL) was added, and the mixture was stirred at room temperature for 0.5 hours. The mixture was concentrated under reduced pressure, and the crude product was separated and purified by Pre-TLC to obtain compound 2 (32 mg, purity 98.5%, yield 56.7%). ¹ H NMR (500 MHz, DMSO-d ₆ ) δ7.25 (d, J＝8.6 Hz, 1H), 6.80 (t, J＝8.5 Hz, 1H), 5.25-5.17 (m, 2H), 5.00 (m, 1H), 4.80 (d, J＝13.8 Hz, 1H), 4.75-4.60 (m, 2H), 4.63 (d, J＝13.8 Hz, 1H), 4.45-4.40 (m, 3H) ,4.15(d,J＝14.2Hz,2H),4.12-4.07(m,3H),3.90(d,J＝14.6Hz,1H),3.69-3.50(m,3H),3.23-3.10(m,3H),2.84(m,2H),2.67(m,1H),2.20-1.84(m, 7H),1.80(m,1H). ESI-MSm/z=573[M+H] ⁺ .

Example 63: Synthesis of Compound 4-(4-((1R,5S)-3,8-diazabicyclo[3.2.1]oct-6-en-3-yl)-8-fluoro-2-((2-methyltetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)pyrido[4,3d]pyrimidin-7-yl)-5-ethynyl-6-fluoronaphthalen-2-ol

Step 1: Synthesis of compound 63-1

Compound M14 (43 mg) was dissolved in N,N-dimethylformamide (2 mL), cesium fluoride (153.72 mg) was added, and the mixture was reacted at room temperature for 3 h. After the reaction was complete, the reaction solution was diluted with EA and water, extracted twice with EA, washed three times with saturated brine, and purified by Pre-TLC to obtain compound 63-1 (19 mg, yield 54.20%). ESI-MS m/z: 693.47 [M+H] +.

Step 2: Synthesis of compound 63

Compound 63-1 (19 mg) was dissolved in dichloromethane (1 mL), and boron trifluoride etherate (0.069 mL) was added and reacted at room temperature for 0.5 h. After the reaction was complete, the reaction solution was added to a cold saturated sodium carbonate solution, extracted twice with DCM, dried, filtered, and spin-dried, and purified by Pre-TLC to obtain compound 63 (11.4 mg, yield 70.02%). ESI-MS m/z: 593.44 [M+H] +. ¹ H NMR(500MHz,MeOD)δ9.05(s,1H),7.85(dd,J＝9.1,5.7Hz,1H),7.37–7.26(m,2H),7.20(d,J＝2.5Hz,1H),6.31(dd,J＝5.9,4.3Hz,2H),5.05(s,2H),4.54(dd,J＝2 6.7,12.4Hz,2H),4.37–4.26(m,2H ),4.12–4.00(m,2H),3.86(dd,J＝18.2,13.2Hz,3H),3.46(d,J＝13.9Hz,1H),3.36(d,J＝3.6Hz,1H),3.32–3.25(m,3H),2.54(d,J＝15.9Hz,1H),2.24–2.14 (m,1H),2.07–1.84(m,3H),1.28(s,1H).

Example 88: Synthesis of Compound 4-(4-((1R,5S)-3,8-diazabicyclo[3.2.1]oct-6-en-3-yl)-8-fluoro-2-((2-methyltetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)pyrido[4,3d]pyrimidin-7-yl)-5-ethynyl-6-fluoronaphthalene-2-amine

Step 1: Synthesis of compound 88-1

Intermediate M14 (520 mg), DMAP (7.48 mg), triethylamine (0.26 mL) were dissolved in tetrahydrofuran (6 mL), and finally 1,1,1-trifluoro-N-phenyl-N-(trifluoromethylsulfonyl)methanesulfonamide (437.56 mg) was added and reacted at room temperature for 1 h. After the reaction was complete, the reaction solution was diluted with EA and water, extracted twice with EA, and purified by column chromatography to obtain compound 88-1 (590 mg, yield 98.33%). ESI-MS m/z: 981.57 [M+H] ⁺ .

Step 2: Synthesis of compound 88-2

88-1 (550 mg), benzophenone imine (203.20 mg), Cs ₂ CO ₃ (547.96 mg), Pd(OAc) ₂ (12.60 mg), BINAP (34.93 mg) were dissolved in 1,4-dioxane (10 mL), replaced with nitrogen, and reacted at 100°C for 1 h. After the reaction was complete, the reaction solution was diluted with EA and water, extracted twice with EA, and purified by column chromatography to obtain compound 88-2 (411 mg, yield 72.42%). ESI-MS m/z: 506.75 [M+H]+.

Step 3: Synthesis of compound 88-3

Compound 88-2 (411 mg) was dissolved in tetrahydrofuran (8 mL), and 2N dilute hydrochloric acid (4 ml) was added to react at room temperature for 10 minutes. After the reaction was complete, the temperature was lowered to 0°C, and the compound was freed with saturated sodium carbonate, extracted twice with EA, washed once with saturated brine, dried, filtered, and concentrated. The crude product was directly used in the next step to obtain compound 88-3 (394 mg). ESI-MS m/z: 848.65 [M+H] ⁺ .

Step 4: Synthesis of compound 88-4

Compound 88-3 (394 mg) was dissolved in N,N-dimethylformamide (5 mL), cesium fluoride (1400.82 mg) was added, and the mixture was reacted at room temperature for 3 h. After the reaction was complete, the reaction solution was diluted with EA and water, extracted twice with EA, washed three times with saturated brine, and purified by column chromatography to obtain compound 88-4 (245 mg, yield 76.80%). ESI-MS m/z: 692.50 [M+H] ⁺ .

Step 5: Synthesis of Compound 88

Compound 88-4 (245 mg) was dissolved in dichloromethane (5 mL), and boron trifluoride etherate (0.89 mL) was added and reacted at room temperature for 0.5 h. After the reaction was complete, the reaction solution was added to a cold saturated sodium carbonate solution, extracted twice with DCM, dried, filtered, and spin-dried, and purified by Pre-TLC to obtain compound 88 (136.10 mg, yield 64.91%). ESI-MS m/z: 592.46 [M+H] ⁺ . ¹ H NMR (500MHz, DMSO) δ9.06(s,1H),7.77(dd,J＝9.1,5.9Hz,1H),7.33(t,J＝9.0Hz,1H),7.03(dd,J＝11.8,2.2Hz,2H),6.34-6.23(m,2H),5.63(s,2H),4.90(s,2H ),4.39(d,J＝9.7Hz,1H),4.23(d,J＝10.5Hz,1H),4.00(d,J＝10.5Hz,1H),3 .97-3.91(m,3H),3.83(s,1H),3.80(d,J＝11.6Hz,1H),3.70(d,J＝10.4Hz,1H),3.55(d,J＝14.1Hz,1H),3.19(d,J＝14.1Hz,1H),3.03-2.95(m,1H),2.5 7(dd,J＝18.7,9.6Hz,2H),2.35(d,J＝15.4Hz,1H),2.00-1.64(m,4H),1.23(s,1H).

The following examples were synthesized by the above method or by similar methods using corresponding intermediates.

Biological tests

Pharmacological experiment 1: Cell proliferation experiment (AGS)

KRas-G12D mutant tumor cells AGS ( CRL-1739TM) were plated in a low-adsorption 96-well plate at a cell density of 1×10 ³ /well and placed in a cell culture incubator for overnight culture. After the cells adhered to the wall, the test compounds were added to the 96-well plate at final concentrations of 10000, 2000, 400, 80, 16, 3.2, 0.64, 0.128, 0.025, and 0nM (the final concentration of DMSO was 0.5%). After culturing at 37°C for 96h, 50μL Cell-titer GLO working solution was added to each well, and the mixture was shaken and incubated at room temperature for 10min. The luminescence value was read on a multifunctional microplate reader, and the luminescence value data was converted into inhibition percentage. The percentage of cell proliferation inhibition was calculated according to the following formula:

Inhibition percentage = (maximum value - measured value) / (maximum value - Blank) × 100

("Maximum" is from 0.5% DMSO control wells, "Blank" is from blank control wells, and "Measured" is from compound-treated wells).

GraphPad Prism software was used for curve fitting and _IC50 values were obtained.

Table 1

Compound Name AGS IC ₅₀ (nM) 63 5.5 88 8.4

Pharmacological experiment 2: Cell p-ERK detection test

KRas-G12D mutant tumor cells AGS ( CRL-1739 ^TM ) were plated in a 96-well plate at a cell density of 4×10 ⁴ /well and cultured in a cell culture incubator overnight. After the cells adhered to the wall, the test compounds were added to the 96-well plate at final concentrations of 3000nM, 600nM, 120nM, 24nM, 4.8nM, 0.96nM, 0.19nM, and 0.5% DMSO. After culturing for 3h, the lysate in the pERKHTRF Kit (Cisbio) was used to lyse each treated cell sample in the 96-well plate (40ul/well). After sufficient lysis, 16ul/well of protein solution and 4ul of premixed pERK-d2 antibody and pERK-EuCryptate antibody were added to the HTRF detection 96-well plate. After incubation at 4°C overnight, the ratio signal value of 665nM/620nM was read on a multifunctional microplate reader to collect raw data. The p-ERK inhibition percentage was calculated according to the following formula:

Inhibition percentage = (maximum value - measured value) / (maximum value - Blank) × 100

("Maximum" is from 0.5% DMSO control wells, "Blank" is from blank control wells, and "Measured" is from compound-treated wells).

GraphPad Prism software was used for curve fitting and _IC50 values were obtained.

Table 2

Although the present invention has been comprehensively described through its embodiments, it is worth noting that various changes and modifications are obvious to those skilled in the art. Such changes and modifications should be included in the scope of the appended claims of the present invention.

Claims (21)

1. A compound represented by general formula (I), its stereoisomers, tautomers, deuterated substances or pharmaceutically acceptable salts: in, X is selected from a bond, NH, O or S; is selected from a single bond or a double bond; X ₁ is selected from O, (CH ₂ ) _0-3 , C(R ₄ ) _1-2 , C(O) or NR ₄ ; R ₄ are each independently selected from H, halogen, C _1-6 alkyl, substituted C _1-6 alkyl, C _1-6 alkoxy, C _3-6 cycloalkyl or substituted C _3-6 cycloalkyl; _X2 is selected from _CR5 or N; _R5 is selected from absent or H; X ₃ is selected from O, (CH ₂ ) _0-3 , C(O), C(R ₆ ) _1-2 or NR ₆ ; R ₆ are each independently selected from H, amino, substituted amino, cyano, C _1-6 alkyl, substituted C _1-6 alkyl, halogen, C _2-6 alkenyl, substituted C _2-6 alkenyl, C _3-6 cycloalkyl or substituted C _3-6 cycloalkyl; X ₄ is selected from O, NR ₇ , (CH ₂ ) _0-3 , C(R ₇ ) _1-2 or C(O); R ₇ are each independently selected from H, amino, substituted amino, cyano, C _1-6 alkyl, substituted C _1-6 alkyl, C _1-6 alkoxy, C _3-6 cycloalkyl-O-, halogen, C _2-6 alkenyl, substituted C _2-6 alkenyl, C _3-6 cycloalkyl or substituted C _3-6 cycloalkyl; L is selected from a bond, -C _1-3 alkylene-, -C _3-14 cycloalkyl _- C _0-3 alkylene- or -3-14 membered heterocyclyl-C 0-3 alkylene-, wherein the -C _1-3 alkylene-, -C _3-14 cycloalkyl-C _0-3 alkylene- or -3-14 membered heterocyclyl-C _0-3 alkylene- is optionally further substituted with one or more _Ra ; The ring A is selected from C _5-14 cycloalkyl, 5-14 membered heterocyclyl, C _6-18 aryl or 5-18 membered heteroaryl, the C _5-14 cycloalkyl or 5-14 membered heterocyclyl is selected from a monocyclic ring, a condensed ring, a spirocyclic ring or a bridged ring, and the C _5-14 cycloalkyl, 5-14 membered heterocyclyl, C _6-18 aryl or 5-18 membered heteroaryl is optionally further substituted by one or more _Ra ; Selected from in, is a double bond, is selected from Z configuration or E configuration; R ₈ is each independently selected from H, halogen, hydroxyl, cyano, C _1-6 alkyl, C _1-6 haloalkyl, C _3-14 cycloalkyl, 3-14 membered heterocyclyl, C _6-18 aryl or 5-18 membered heteroaryl, wherein the C _1-6 alkyl, C _1-6 haloalkyl, C _3-14 cycloalkyl, 3-14 membered heterocyclyl, C _6-18 aryl or 5-18 membered heteroaryl is optionally further substituted with one or more halogen, hydroxyl, cyano, carboxyl, amino, nitro, C _1-6 alkyl or C _1-6 alkoxy; R ₂ is selected from C _3-14 cycloalkyl, 3-14 membered heterocyclyl, C _6-18 aryl or 5-18 membered heteroaryl, wherein the C _3-14 cycloalkyl, 3-14 membered heterocyclyl, C _6-18 aryl or 5-18 membered heteroaryl is optionally further substituted with one or more _Ra ; _Ra is independently selected from H, hydroxy, cyano, halogen, _C1-6 alkyl, _C1-6 haloalkyl, _C1-6 alkoxy, _-C0-3 alkylene- _ORb , -OC(=O) _C1-6 alkyl, -C0-3 alkylene- _SRb , _-C0-3 alkylene-N( _Rb ) ₂ , _-C0-3 alkylene-S(=O) _Rb , _{-C0-3 alkylene-S(=O)2Rb, -C0-3 alkylene - SRb , -C0-3 alkylene} -S( _Rb ) ₅ , _-C0-3 alkylene-C(=O) _Rb , _-C0-3 alkylene-C(=O) _ORb , _-C0-3 alkylene-C(=O)N( _Rb ) ₂ , _C2-6 alkenyl, _C2-6 alkynyl, -C -C _0-3 alkylene-C _3-14 cycloalkyl, -C _0-3 alkylene-(3-14 membered heterocyclyl), -C _0-3 alkylene-C _6-18 aryl or -C _0-3 alkylene-(5-18 membered heteroaryl), the C _1-6 alkyl, C _{1-6 haloalkyl, C 1-6 alkoxy} , C _2-6 alkenyl, C _2-6 alkynyl, -C _0-3 alkylene-C 3-14 cycloalkyl, -C _0-3 alkylene-( _3-14 membered heterocyclyl), -C _0-3 alkylene-C _6-18 aryl or -C _0-3 alkylene-(5-18 membered heteroaryl) is optionally further substituted with one or more R _b ; each R _b is independently H, halogen, hydroxy, cyano, C _1-6 alkyl, C _3-6 cycloalkyl, C _{1-6 haloalkyl, C 3-14 cycloalkyl} , 3-14 membered heterocyclyl, C _6-18 aryl or 5-18 membered heteroaryl, said C _1-6 alkyl, C _3-6 cycloalkyl, C _1-6 haloalkyl, C _3-14 cycloalkyl, 3-14 membered heterocyclyl, C _6-18 aryl or 5-18 membered heteroaryl optionally further substituted with one or more halogen, C _1-6 alkyl or C _1-6 haloalkyl; R ₃ is selected from H, halogen, -C _0-3 alkylene cyano, C _1-6 alkyl, C _1-6 alkoxy or C _1-6 haloalkyl. 2. A compound represented by general formula (I), its stereoisomers, tautomers, deuterated substances or pharmaceutically acceptable salts, which is selected from compounds represented by formula (IA) to formula (IF), their stereoisomers, tautomers, deuterated substances or pharmaceutically acceptable salts: Wherein, R ₁ , R ₂ , R ₃ , R ₄ , R ₆ , R ₇ , X, L and A have the same definitions as in claim 1. 3. A compound represented by general formula (I), its stereoisomer, tautomer, deuterated product or pharmaceutically acceptable salt, which is selected from the compounds represented by formula (IA-1) to formula (IF-1), its stereoisomer, tautomer, deuterated product or pharmaceutically acceptable salt: Wherein, R ₁ , R ₂ , R ₃ , R ₄ , R ₆ , R ₇ and A have the same definitions as in claim 1. 4. The compound, stereoisomer, tautomer, deuterated substance or pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, characterized in that the ring A is selected from a C _5-14 cycloalkyl group or a 5-14 membered heterocyclic group. 5. The compound according to claim 4, its stereoisomer, tautomer, deuterated substance or pharmaceutically acceptable salt, characterized in that the 5-14 membered heterocyclic group is a condensed ring. 6. The compound according to claim 5, its stereoisomer, tautomer, deuterated substance or pharmaceutically acceptable salt, characterized in that the fused ring is selected from 7. The compound according to any one of claims 1 to 6, its stereoisomer, tautomer, deuterated substance or pharmaceutically acceptable salt, characterized in that: Selected from 8. The compound, stereoisomer, tautomer, deuterated substance or pharmaceutically acceptable salt thereof according to claim 7, characterized in that R ₈ is independently selected from H, halogen, C _1-6 alkyl, C _3-14 cycloalkyl, 3-14 membered heterocyclyl, C _6-18 aryl or 5-18 membered heteroaryl, and the C _3-14 cycloalkyl, 3-14 membered heterocyclyl, C _6-18 aryl or 5-18 membered heteroaryl is optionally further substituted with one or more halogen, hydroxyl, cyano, carboxyl, amino, nitro or C _1-6 alkyl. 9. The compound according to any one of claims 1 to 8, its stereoisomer, tautomer, deuterated substance or pharmaceutically acceptable salt, characterized in that: Selected from 10. The compound according to claim 9, its stereoisomer, tautomer, deuterated substance or pharmaceutically acceptable salt, characterized in that Selected from 11. A compound represented by general formula (I), its stereoisomer, tautomer, deuterated product or pharmaceutically acceptable salt, which is selected from the compounds represented by formula (IA-1-1) to formula (IF-1-1), its tautomer, deuterated product or pharmaceutically acceptable salt: Wherein, R ₁ , R ₂ , R ₃ , R ₄ , R ₆ and R ₇ have the same definitions as in claim 1. 12. The compound, stereoisomer, tautomer, deuterated product or pharmaceutically acceptable salt thereof according to any one of claims 1 to 11, characterized in that _R4 is H or halogen, and the halogen is preferably F. 13. The compound, stereoisomer, tautomer, deuterated substance or pharmaceutically acceptable salt thereof according to any one of claims 1 to 12, characterized in that _R6 is H, alkyl or halogen. 14. The compound, stereoisomer, tautomer, deuterated product or pharmaceutically acceptable salt thereof according to any one of claims 1 to 13, characterized in that _R7 is H, _C1-6 alkoxy or _C3-6 cycloalkyl-O-. 15. A compound, stereoisomer, tautomer, deuterated substance or pharmaceutically acceptable salt thereof according to any one of claims 1 to 14, characterized in that R ₂ is selected from C _6-18 aryl or 5-18 membered heteroaryl, wherein the C _6-18 aryl or 5-18 membered heteroaryl is optionally further substituted with one or more _Ra , wherein _Ra is independently selected from H, hydroxyl, cyano, amino, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _2-6 alkenyl, C _2-6 alkynyl or -C _0-3 alkylene-C _3-14 cycloalkyl. 16. The compound according to any one of claims 15, its stereoisomer, tautomer, deuterated substance or pharmaceutically acceptable salt, characterized in that R ₂ is selected from Said Optionally further substituted _with one or more _Ra independently selected from H, hydroxy, cyano, halogen, amino, _C1-6 alkyl, _C1-6 alkoxy, _C2-6 alkenyl, _C2-6 alkynyl or _-C0-3 alkylene- _C3-14 cycloalkyl. 17. The compound according to any one of claims 16, its stereoisomer, tautomer, deuterated substance or pharmaceutically acceptable salt, characterized in that R ₂ is selected from 18. The compound, stereoisomer, tautomer, deuterated substance or pharmaceutically acceptable salt thereof according to any one of claims 1 to 17, characterized in that _R3 is H, methoxy or halogen. 19. The compound according to any one of claims 1 to 18, its stereoisomer, tautomer, deuterated substance or pharmaceutically acceptable salt, characterized in that formula (I) is selected from 20. A pharmaceutical composition, characterized in that the pharmaceutical composition contains a therapeutically effective amount of the compound according to any one of claims 1 to 19 or its stereoisomer, tautomer, deuterated substance or pharmaceutically acceptable salt. 21. Use of the compound according to any one of claims 1 to 19 or its stereoisomer, tautomer, deuterated substance or pharmaceutically acceptable salt or the pharmaceutical composition according to claim 20 in the preparation of a medicament for a disease mediated by KRAS G12D.