Aromatic amide derivative and application thereof in antitumor drugs

Abstract

The invention provides aromatic amide derivatives, stereoisomers, tautomers, solvates and pharmaceutically acceptable salts thereof, and also provides a preparation method of the compounds and application of the compounds to prevention and / or treatment of cell proliferative diseases, especially MEK, tautomers, solvates and pharmaceutically acceptable salts thereof, alone or in combination with other drugs. The invention also relates to applications in the aspects of cancer-related diseases mediated by Raf.

Description

Aromatic amide derivative and its application in anti-tumor drugs Technical Field

[0001] The present invention belongs to the field of medicinal chemistry and relates to an aromatic amide derivative, a preparation method thereof and application of the compound in treating proliferative diseases. Background Art

[0002] Aromatic amide compounds are widely present in drug molecular structures and can serve as pharmacophores that interact with drug target proteins, playing a significant role in improving drug performance. For example, the following marketed drugs or candidate drugs in clinical research all contain aromatic amide structures in their molecular structures and exhibit a wide range of pharmacological effects.

[0003] The RAS-RAF-MEK-ERK signaling pathway transmits extracellular signals to specific intracellular targets and plays a crucial role in cell proliferation, survival, and differentiation. Stimulated by upstream molecules such as cytokines, RAS transitions from an inactive to an active conformation, thereby recruiting intracellular RAF to the cell membrane and promoting its dimerization and phosphorylation. Activated RAF, in turn, phosphorylates and activates MEK and ERK, ultimately regulating cell proliferation, differentiation, apoptosis, and metastasis.

[0004] RAS, a crucial upstream component of this pathway, acts as a molecular switch by binding to GTP / GDP. RAS includes three isoforms: KRAS, NRAS, and HRAS. KRAS is the most frequently mutated in human tumors, including approximately 90% of pancreatic cancers, 45% of colorectal cancers, and 35% of lung cancers (Cancer Discov, 2022, 12:924). RAF, including three isoforms: ARAF, BRAF, and CRAF, is a downstream kinase of RAS. RAS-GTP can directly bind to a serine / threonine kinase, RAF, activating RAF. Activated RAF kinase phosphorylates MEK. MEK1 / 2, a downstream kinase of RAF, is a dual-specificity kinase that catalyzes the tyrosine and serine / threonine residues of ERK1 / 2. ERK functions in the cytoplasm and nucleus, phosphorylating hundreds of substrates, including transcription factors and protein kinases (Genes and Diseases, 2023, 10:76).

[0005] Abnormal activation of the RAS-RAF-MEK-ERK signaling pathway contributes to the development and progression of various tumors. Frequent mutations in the RAS-RAF-MEK-ERK pathway have been observed in human cancers, making targeting this pathway an effective therapeutic strategy for mitigating tumor growth. Approximately 33% of tumors harbor RAS mutations, and 8% harbor RAF mutations. MEK and ERK mutation frequencies in human tumors are relatively low, but high levels of MEK and ERK activity are found in over 85% of cancers (Genes and Diseases, 2023, 10:76). To date, various single-target inhibitors have been developed, including the KRAS inhibitors sotorasib and adagrasib, which are currently marketed or in clinical trials; the BRAF inhibitors vemurafenib, dabrafenib, and encorafenib; the MEK inhibitors trametinib, binimetinib, and cobimetinib; and the ERK inhibitor GDC-0994. However, studies have shown that single-target inhibitor drugs often have disadvantages such as high drug resistance, severe toxic side effects, and narrow indications, which are insufficient to achieve ideal anti-tumor effects. Currently, RAF and MEK inhibitors are used in combination to block the two key targets of this pathway in clinical practice to overcome drug resistance or delay the development of drug resistance.

[0006] Developing dual-target RAF / MEK inhibitors is an effective anti-tumor strategy. Among the RAF / MEK inhibitors currently under investigation (relevant documents include WO2021149776 and WO2007091736), VS-6766 has made rapid clinical progress, demonstrating efficacy in treating various KRAS-mutant tumors, with associated adverse reactions also reported in clinical trials. Developing such inhibitors with both high efficacy and safety is of great significance.

[0007] Summary of the Invention

[0008] The present invention discloses an aromatic amide derivative, and its stereoisomers, tautomers or pharmaceutically acceptable salts, as MEK / RAF dual-target inhibitors, have the advantages of high activity and good safety, and show good application prospects in preventing and / or treating cell proliferative diseases related to the MAPK pathway, especially tumors.

[0009] The present invention provides a compound represented by formula (I), its stereoisomers, tautomers, solvates and pharmaceutically acceptable salts.

[0010] Wherein, U1, U2, U3, U4 are each independently selected from nitrogen, CR U ;

[0011] RU Selected from R 5c , R 5d , R 6b , R 6c ;

[0012] W1, W2, and W3 are each independently a bond, NR W ;

[0013] R W is selected from hydrogen, alkyl, deuterated alkyl;

[0014] Y is selected from oxygen or sulfur;

[0015] R1 is selected from -S(O)2NH-R7 or -S(O)2-R7;

[0016] R7 is selected from hydrogen, substituted or unsubstituted alkyl, deuterated alkyl, haloalkyl, alkoxyalkyl, alkoxy, cycloalkyl, alkenyl, alkynyl;

[0017] R2 is selected from hydrogen, substituted or unsubstituted alkyl, deuterated alkyl, haloalkyl, alkoxy, alkoxyalkyl, alkenyl, alkynyl, cycloalkyl;

[0018] R 3a , R 3b , R 4a , R 4b , R 4c , R 4d , R 4e , R 5a , R 5b , R 5c , R 5d , R 6a , R 6b , R 6c Each is independently selected from hydrogen, deuterium, halogen, hydroxyl, substituted or unsubstituted amino, alkyl, deuterated alkyl, haloalkyl, alkoxy, alkenyl, alkynyl, alkylthio, cycloalkyl, heterocyclic group, aryl, heteroaryl, or two adjacent substituents can together form a substituted or unsubstituted cyclic structure.

[0019] The present invention provides a compound represented by formula (II), its stereoisomers, tautomers, solvates and pharmaceutically acceptable salts.

[0020] Wherein, U1 is selected from nitrogen, CR 5d ;

[0021] R7 is selected from hydrogen, substituted or unsubstituted alkyl, deuterated alkyl, haloalkyl, alkoxyalkyl, alkoxy, cycloalkyl, alkenyl, alkynyl;

[0022] R2 is selected from hydrogen, substituted or unsubstituted alkyl, deuterated alkyl, haloalkyl, alkoxy, alkoxyalkyl, alkenyl, alkynyl, cycloalkyl;

[0023] R 3a , R 3b , R 4a , R 4b , R 4c , R 4d , R 4e , R 5a , R 5b , R 5c , R 5d , R 6a , R 6b , R 6c Each is independently selected from hydrogen, deuterium, halogen, hydroxyl, substituted or unsubstituted amino, alkyl, deuterated alkyl, haloalkyl, alkoxy, alkenyl, alkynyl, alkylthio, cycloalkyl, heterocyclic group, aryl, heteroaryl, or two adjacent substituents can together form a substituted or unsubstituted cyclic structure.

[0024] The present invention provides a compound represented by formula (III), its stereoisomers, tautomers, solvates and pharmaceutically acceptable salts.

[0025] Wherein, U1 is selected from nitrogen, CR 5d ;

[0026] R7 is selected from hydrogen, substituted or unsubstituted alkyl, deuterated alkyl, haloalkyl, alkoxy, alkenyl, alkynyl, cycloalkyl;

[0027] R 8a , R 8b , R 8c Each is independently selected from hydrogen, deuterium, halogen, hydroxy, substituted or unsubstituted alkyl, deuterated alkyl, haloalkyl, cycloalkyl, heterocyclyl;

[0028] R 3a , R 3b , R 4a , R 4b , R 4c , R 4d , R 4e , R 5a , R 5b , R 5c , R 5d , R 6a , R 6b , R 6cEach is independently selected from hydrogen, deuterium, halogen, hydroxyl, substituted or unsubstituted amino, alkyl, deuterated alkyl, haloalkyl, alkoxy, alkenyl, alkynyl, alkylthio, cycloalkyl, heterocyclyl, aryl, heteroaryl, or two adjacent substituents may together form a substituted or unsubstituted cyclic structure;

[0029] n is 1, 2, 3, 4.

[0030] The present invention provides the following compounds, their stereoisomers, tautomers, solvates and pharmaceutically acceptable salts,

[0031] The present invention provides a compound represented by formula (IV), its stereoisomers, tautomers or pharmaceutically acceptable salts,

[0032] Wherein, U1 is selected from nitrogen or CR 5d ;

[0033] R 5d Selected from hydrogen or C 1-6 alkyl;

[0034] W is selected from a chemical bond or oxygen;

[0035] R 4a , R 4b , R 4c , R 4d , R 4e are independently selected from hydrogen, halogen, aldehyde, C 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 Alkenyl, C 2-6 Alkynyl or C 3-6 Cycloalkyl;

[0036] R 5a , R 5b , R 5c , R 6a , R 6b , R 6c are each independently selected from hydrogen, halogen or C 1-6 alkyl;

[0037] R7 is selected from hydrogen, C 1-6 Alkyl, C 1-6 Deuterated alkyl or C 3-6 Cycloalkyl;

[0038] R 8a , R 8b , R 8c are each independently selected from hydrogen, deuterium, C 1-6 Alkyl or C 3-6 Cycloalkyl;

[0039] n is 1, 2, 3, 4.

[0040] The present invention provides a compound represented by formula (V), its stereoisomers, tautomers or pharmaceutically acceptable salts,

[0041] wherein U1 is selected from nitrogen or CH;

[0042] R 4a is selected from hydrogen or halogen;

[0043] R 4c Selected from hydrogen, halogen, aldehyde, C 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 Alkenyl, C 2-6 Alkynyl or C 3-6 Cycloalkyl;

[0044] R7 is selected from C 1-6 Alkyl, C 1-6 Deuterated alkyl or C 3-6 Cycloalkyl;

[0045] R 8a , R 8b are each independently selected from hydrogen, deuterium or C 1-6 alkyl;

[0046] R 8c Selected from hydrogen or C 1-6 alkyl;

[0047] n is 1, 2, 3.

[0048] The present invention provides the following compounds, their stereoisomers, tautomers or pharmaceutically acceptable salts,

[0049] The present invention provides a compound represented by formula (VI), its stereoisomers, tautomers or pharmaceutically acceptable salts,

[0050] wherein U1 is selected from nitrogen or CH;

[0051] R 4a is selected from hydrogen or halogen;

[0052] R 4c Selected from hydrogen, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 Alkenyl or C 2-6 Alkynyl;

[0053] R7 is selected from C1-6 Alkyl, C 1-6 Deuterated alkyl or C 3-6 Cycloalkyl;

[0054] R 8a , R 8b are each independently selected from hydrogen, deuterium or C 1-6 alkyl;

[0055] R 8c Selected from hydrogen, C 1-6 Alkyl or C 3-6 Cycloalkyl;

[0056] n is 1, 2, 3.

[0057] The present invention provides the following compounds, their stereoisomers, tautomers or pharmaceutically acceptable salts,

[0058] The present invention provides a pharmaceutical composition comprising a therapeutically effective dose of any one of the compounds of the present invention, or a stereoisomer, tautomer, solvate or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0059] Use of any one of the compounds of the present invention or its pharmaceutically acceptable salt or the pharmaceutical composition of the present invention in the preparation of a medicament for preventing and / or treating diseases related to MEK and Raf-mediated cancers, wherein the use comprises using any one of the compounds of the present invention or its pharmaceutically acceptable salt or the pharmaceutical composition of the present invention alone or in combination with other treatment methods including immunotherapy to prevent and / or treat diseases related to MEK and Raf-mediated cancers.

[0060] Use of any one of the compounds of the present invention or its pharmaceutically acceptable salt or the pharmaceutical composition of the present invention in the preparation of a method for preventing and / or treating a cell proliferative disease, wherein the use includes using any one of the compounds of the present invention or its pharmaceutically acceptable salt or the pharmaceutical composition of the present invention alone or in combination with other treatment methods including immunotherapy to prevent and / or treat a cell proliferative disease.

[0061] The use of the present invention, wherein the diseases include but are not limited to renal cancer, liver cancer, breast cancer, pancreatic cancer, prostate cancer, melanoma, leukemia, malignant lymphoma, ovarian cancer, head and neck cancer, lung cancer, colon cancer, bladder cancer, uterine cancer, etc.

[0062] Detailed Description of the Invention

[0063] All technical and scientific terms used in this specification have the same meanings as commonly understood by those skilled in the art.

[0064] The term "hydrogen" refers herein to -H.

[0065] The term "deuterium" refers herein to -D.

[0066] The term "oxygen" refers to O herein.

[0067] The term "sulfur" refers to S herein.

[0068] The term "nitrogen" refers to N herein.

[0069] The term "halogen" as used herein refers to -F, -Cl, -Br and -I.

[0070] The term "hydroxy" refers herein to -OH.

[0071] The term "aldehyde group" refers herein to -CHO.

[0072] The term "amino" or "amine" refers interchangeably to a -NR2 group, wherein each R is, for example, H or a substituent. In some embodiments, the amino group is further substituted to form an ammonium ion, for example, NR 3+ Ammonium moieties are specifically included in the definition of "amino" or "amine." Substituents can be, for example, alkyl, deuterated alkyl, alkoxy, cycloalkyl, heterocycloalkyl, amide, or carboxylate. The R group can be further substituted with one or more (e.g., 1 to 4) groups selected from the group consisting of halo, cyano, alkenyl, alkynyl, alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, urea, carbonyl, carboxylate, amine, and amide.

[0073] The term "alkyl" refers to a saturated aliphatic hydrocarbon group having 1 to 10 carbon atoms in this article, and the term includes straight and branched chain hydrocarbon groups. Non-limiting examples of alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, neopentyl, n-hexyl, etc. Alkyl described herein can be optionally substituted with one or more of the following substituents: deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, hydroxyl, carboxyl, amino, alkyl, alkoxy, acyl, acyloxy, oxo, amide, ester, amido, cycloalkyl, cycloalkenyl, heterocycloalkyl, alkenyl, alkenyloxy, alkynyl, cycloalkyloxy, heterocycloalkyloxy, aryloxy, heteroaryloxy, aryl or heteroaryl.

[0074] The term "deuterated alkyl" as used herein refers to an alkyl group in which the aforementioned "alkyl" group is substituted with deuterium.

[0075] The term "haloalkyl" as used herein refers to an alkyl group obtained by substituting the aforementioned "alkyl" group with a halogen.

[0076] The term "alkoxy" refers to a group of the formula -OR a Group, where Ra is an alkyl group as defined above. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, isopropoxy, and the like.

[0077] The term "alkoxyalkyl" herein refers to an alkyl group obtained by substituting the aforementioned "alkyl" group with the aforementioned "alkoxy" group.

[0078] The term "alkylthio" as used herein refers to a group of the formula -SR a Group, where R a is an alkyl group as defined above. Examples of alkylthio groups include, but are not limited to, methylthio, ethylthio, isopropylthio, and the like.

[0079] Alkenyl is an unsaturated hydrocarbon group containing a carbon-carbon double bond. The term "alkenyl" refers to an alkyl group containing a carbon-carbon double bond in the molecule in this article, wherein the alkyl group is as defined above. Alkenyl described herein can be optionally substituted with one or more of the following substituents: deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, hydroxyl, carboxyl, amino, alkyl, alkoxy, acyl, amide, ester group, amido, sulfonyl, sulfinyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, cycloalkyloxy, sulfydryl, alkyl mercapto, deuterated alkyl mercapto, sulfone, sulfoxide, amino, silicon, phosphono, deuterated alkyl, heterocycloalkyl, aryl, heteroaryl, alkynyl, alkenyl, arylalkyl, ester group. Non-limiting examples of alkenyl include vinyl, propenyl, allyl, isopropenyl, butenyl, isobutenyl etc.

[0080] Alkynyl is an unsaturated hydrocarbon radical containing a carbon-carbon triple bond. Term " alkynyl " refers to the alkyl group containing a carbon-carbon triple bond in the molecule in this article, wherein the alkyl is as defined above. Alkynyl can be substituted or unsubstituted, and when substituted, substituent is preferably one or more following groups, which are independently selected from deuterium, fluorine, chlorine, bromine, iodine, cyano group, nitro, hydroxyl, carboxyl, amino, alkyl, alkoxy, acyl, amide, ester group, amido, sulfonyl, sulfinyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, cycloalkyloxy, sulfydryl, alkyl sulfydryl, deuterated alkyl sulfydryl, sulfone, sulfoxide, amino, silicon-based, phosphono, deuterated alkyl, heterocycloalkyl, aryl, heteroaryl, alkynyl, alkenyl, arylalkyl, ester group. Non-limiting examples of alkynyl include ethynyl, 1-propynyl, 2-propynyl, 1-, 2- or 3-butynyl etc.

[0081] The term "cycloalkyl" means a stable monovalent non-aromatic monocyclic or polycyclic hydrocarbon radical consisting solely of carbon and hydrogen atoms, which may include fused or bridged ring systems, having 3 to 15 carbon atoms, preferably 3 to 10 carbon atoms, more preferably 3 to 8 carbon atoms, and which is saturated or unsaturated and may be attached to the rest of the molecule by a single bond via any suitable carbon atom. Cycloalkyl includes, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 1H-indenyl, 2,3-dihydroindanyl, 1,2,3,4-tetrahydro-naphthyl, 5,6,7,8-tetrahydro-naphthyl, 8,9-hydro-7H-benzocyclohepten-6-yl, 6,7,8,9-tetrahydro-5H-benzocycloheptenyl, 5,6,7,8,9,10-hexahydro-benzocyclooctenyl, fluorenyl, bicyclo [2.2.1]heptyl, 7,7-dimethyl-bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, bicyclo[2.2.2]octyl, bicyclo[3.1.1]heptyl, bicyclo[3.2.1]octyl, bicyclo[2.2.2]octenyl, bicyclo[3.2.1]octenyl, adamantyl, octahydro-4,7-methylene-1H-indenyl and octahydro-2,5-methylene-pentalenyl, and the like.

[0082] The term "heterocyclyl" refers to a substituted or unsubstituted saturated or unsaturated aromatic or non-aromatic ring containing at least 1 to 5 heteroatoms selected from nitrogen, oxygen, or sulfur. Aromatic and non-aromatic rings may be 3- to 10-membered monocyclic rings, 4- to 20-membered spirocyclic rings, cyclic rings, or bridged rings. The nitrogen and sulfur atoms optionally substituted in the heterocyclyl ring may be oxidized to various oxidation states. A 3- to 12-membered heterocyclic ring is preferred. Non-limiting examples include oxirane, oxetanyl, oxolanyl, oxhexyl, oxepanyl, oxocanyl, aziridinyl, azetidinyl, azirol, azihexyl, azirinyl, 1,3-dioxolanyl, 1,4-dioxolanyl, 1,3-dithiolanyl, 1,3-dioxohexanyl, 1,3-dithiohexanyl, azepinyl, morpholinyl, piperazinyl, pyridinyl, furanyl, thienyl, pyrrolyl, pyranyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, piperidinyl, thiomorpholinyl, dihydropyran, thiadiazolyl, oxazolyl, oxadiazolyl, pyrazolyl, 1,4-dioxadienyl, and the like.

[0083] The term "aryl" herein refers to a 6-10 membered all-carbon monocyclic or fused polycyclic (i.e., rings that share adjacent pairs of carbon atoms) group, a polycyclic (i.e., rings with adjacent pairs of carbon atoms) group having a conjugated π electron system. The aryl group can be covalently attached to the defined chemical structure at any carbon atom that produces a stable structure. The aryl groups described herein may be optionally substituted with one or more of the following substituents: deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, hydroxyl, carboxyl, amino, alkyl, alkoxy, acyl, amide, ester, amine, sulfonyl, sulfinyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, alkenyl, alkynyl, and cycloalkoxy.

[0084] The term "heteroaryl" as used herein refers to an aromatic group consisting of 5 to 10 atoms and containing at least one heteroatom selected from N, O, or S. The term can have a single ring (non-limiting examples include furan, thiophene, imidazole, pyrazole, pyridine, pyrazine, oxazole, thiazole, etc.) or multiple fused rings (non-limiting examples include benzothiophene, benzofuran, indole, isoindole, etc.), wherein the fused rings may or may not be aromatic groups containing heteroatoms, provided that the point of attachment is through an atom of the aromatic heteroaryl group. The heteroaryl groups described herein may be optionally substituted with one or more of the following substituents: deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, hydroxy, amino, alkyl, alkoxy, acyl, acyloxy, amide, ester, amine, sulfonyl, sulfinyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, alkenyl, alkynyl, and cycloalkoxy.

[0085] The term "amide" (or "amido") includes C-amide groups and N-amide groups, ie, -C(O)NR A R B and -NR A C(O)R B Group. R A and R B is independently hydrogen or substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl or heterocyclyl as defined herein. Amide groups therefore include, but are not limited to, carbamoyl (-C(O)NH2) and formamide groups (-NHC(O)H). In some embodiments, an amide is -NR A C(O)-(C 1-5 In some embodiments, the amide is -NHC(O)-alkyl), which group is referred to as a "carbonylamino" group. In other embodiments, the amide is -NHC(O)-alkyl, which group is referred to as an "alkanoylamino" group.

[0086] The term "sulfonamide" includes both S-sulfonamide and N-sulfonamide groups, i.e., -SO2NR C R D and -NR C SO2R D Group. RC and R D is independently hydrogen or substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl or heterocyclyl as defined herein. Sulfonamide groups therefore include, but are not limited to, sulfonyl (-SO2NH2). In some embodiments herein, sulfonamide is -NHSO2-alkyl, which is referred to as "alkylsulfonylamino".

[0087] The present invention also includes isotopically labeled compounds of the present invention, i.e., compounds having the same structure as disclosed above, but with one or more atoms in the structure replaced by atoms having the same number of protons but a different number of neutrons. Examples of isotopes that may be incorporated into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, sulfur, fluorine, chlorine, and iodine, such as 2 H, 3 H, 13 C, 14 C, 15 N, 18 O, 17 O, 35 S, 18 F, 36 Cl and 131 I, etc. The compounds of the present invention, their stereoisomers, tautomers or pharmaceutically acceptable salts, and compounds of the above forms containing the above isotopes and / or other atomic isotopes, are all within the scope of the present invention. Certain isotopically labeled compounds of the present invention, such as 3 H or 14 Compounds labeled with C can be used in drug tissue distribution studies. 3 H or 14 C isotopes are particularly preferred due to their ease of preparation and detection. 2 H, 18 O replaces some of the compounds of the present invention due to better metabolic stability and have certain therapeutic advantages, such as increased half-life in vivo and lower dosage and other comprehensive properties, therefore, 2 H, 18 O is also preferred in some cases.

[0088] The term "compound of the present invention" (unless otherwise specifically stated) refers to the compounds of formula (I) to formula (III) and all pure and mixed stereoisomers, geometric isomers, tautomers, solvates, hydrates, prodrugs and isotopically labeled compounds and any pharmaceutically acceptable salts thereof. A solvate of a compound of the present invention refers to a compound or salt thereof in combination with a stoichiometric or non-stoichiometric amount of a solvent, such as a hydrate, ethanolate, methanolate, etc. The compound may also exist in one or more crystalline states, i.e., as a co-crystal, polymorph, or it may exist as an amorphous solid. All such forms are encompassed by the claims.

[0089] The term "pharmaceutically acceptable" means that the substance or composition must be compatible chemically and / or toxicologically with the other ingredients comprising the formulation and / or the mammal to be treated therewith.

[0090] The term "stereoisomer" as used herein refers to compounds having one or more stereocenters that differ in chirality. Stereoisomers include enantiomers, diastereomers, conformers (rotamers), geometric isomers (cis / trans) isomers, atropisomers, and the like.

[0091] The compounds of the present invention may be used in the form of salts, such as "pharmaceutically acceptable salts" derived from inorganic or organic acids. These include, but are not limited to, acetate, adipate, alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate, digluconate, cyclopentanepropionate, sodium dodecylbenzenesulfonate sulfate, ethanesulfonate, glucose heptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, fumarate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, hydrochloride, 2-naphthalenesulfonate, oxalate, pectinate, sulfate, 3-phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, p-toluenesulfonate, and quinate.

[0092] In this application, "cell proliferative disease" refers to a disease caused by abnormal cell division and proliferation, particularly cancer. In this application, "cancer" includes, but is not limited to, blood and lymphatic cancers such as leukemia, malignant lymphoma, multiple myeloma, and myelodysplastic syndrome; and solid tumors such as brain tumors, neurological tumors, head and neck cancer, esophageal cancer, gastric cancer, colorectal cancer, lung cancer, thyroid cancer, breast cancer, gallbladder cancer, pancreatic cancer, liver cancer, prostate cancer, ovarian cancer, uterine cancer, testicular cancer, ovarian cancer, renal cell carcinoma, bladder cancer, renal pelvis-ureter cancer, malignant melanoma, and skin cancer.

[0093] The term "compound of the present invention" (unless otherwise specifically stated) refers to the compounds of formula (IV) to formula (VI) and all pure and mixed stereoisomers, geometric isomers, tautomers, solvates, hydrates, prodrugs and isotopically labeled compounds and any pharmaceutically acceptable salts thereof. Solvates of the compounds of the present invention refer to compounds or salts thereof combined with stoichiometric and non-stoichiometric amounts of solvents, such as hydrates, ethanolates, methanolates, etc. The compounds may also exist in one or more crystalline states, i.e., as co-crystals, polymorphs, or they may exist as amorphous solids. All such forms are encompassed by the claims.

[0094] The term "optional" or "optionally" means that the subsequently described event or circumstance can but need not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0095] At various places in this specification, substituents of the compounds of the invention are disclosed in the form of groups or ranges. This specifically means that the invention includes each member of such groups and ranges or each individual subcombination of the members. For example, the term "C 1-6 "Alkyl" specifically means methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl and C6 alkyl are disclosed individually.

[0096] The term "substituted or unsubstituted ..." means that all radical structures following the term are unsubstituted or substituted by one or more substituents described in the present invention. The term "substituted" in this article means that any group is monosubstituted or polysubstituted by a specified substituent to the extent that such monosubstituted or polysubstituted (including multiple substitutions in the same part) is chemically permitted, and each substituent can be located at any available position on the group and can be connected by any available atom on the substituent. "Any available position" refers to any position on the group that can be chemically obtained by methods known in the art or methods taught herein, and does not produce excessively unstable molecules. When there are two or more substituents on any group, each substituent is defined independently of any other substituents and therefore can be the same or different.

[0097] The term "tautomer" as used herein refers to structural isomers with different energies that can interconvert across a low energy barrier. Examples include proton tautomers, such as enol-keto tautomers and imine-enamine tautomers, or tautomeric forms of heteroaryl groups containing ring atoms attached to both the ring-NH- portion and the ring=N- portion, such as pyrazole, imidazole, benzimidazole, triazole, and tetrazole. Valence tautomers include interconversion by reorganization of some of the bonding electrons.

[0098] In this application, "tumor" includes but is not limited to: leukemia, malignant lymphoma, multiple myeloma, myelodysplastic syndrome and other blood and lymphatic cancers; as well as brain tumors, nerve tumors, head and neck cancer, esophageal cancer, gastric cancer, colorectal cancer, lung cancer, thyroid cancer, breast cancer, gallbladder cancer, pancreatic cancer, liver cancer, prostate cancer, ovarian cancer, uterine cancer, testicular cancer, ovarian cancer, renal cell carcinoma, bladder cancer, renal pelvis-ureter cancer, malignant melanoma, skin cancer and other solid tumors. DETAILED DESCRIPTION

[0099] The present invention is further illustrated by the following examples, but the present invention is not limited thereto. Throughout this application, various examples of the compounds and methods of the present invention are mentioned herein. The present invention is not limited to these examples. The following examples are merely provided to provide methods for practicing the present invention and are not intended to limit the scope of the present invention in any way.

[0100] The compounds provided herein can be prepared by standard synthetic methods known in the art, and this specification provides general methods for preparing the compounds of the present invention. Starting materials can generally be obtained commercially or prepared by methods well known to those skilled in the art.

[0101] The preparation process of general formula II is as follows:

[0102] Under alkaline conditions, an aniline derivative b and a fluorobenzene derivative a are reacted to form a compound c; a benzoic acid derivative c is reacted with a methylating agent to form a compound d; an aldehyde derivative d is reacted with an arylsulfonylhydrazine to form a compound e; under alkaline conditions, a hydrazine derivative e and an arylboronic acid derivative f are reacted to form a compound g; under acidic conditions, compound g is deprotected to form a compound h; an ester derivative h is hydrolyzed in the presence of a hydroxide to form a compound i; when a condensing agent is present, the benzoic acid derivative i is reacted with a corresponding amine or amine hydrochloride to form a compound j; and under alkaline conditions, compound j is reacted with a corresponding sulfonyl to prepare a compound of formula II.

[0103] or

[0104] Compound h is reacted with the corresponding sulfonyl to form compound k, and the ester derivative k is hydrolyzed in the presence of a hydroxide to form compound l; when a condensing agent is present, the benzoic acid derivative l reacts with the corresponding amine or amine hydrochloride to obtain a compound of formula II.

[0105] The compounds of the present invention and corresponding preparation methods are further explained and listed below by examples and preparations. It should be understood that although typical or preferred reaction conditions are given in the specific examples, those skilled in the art may also use other reaction conditions. Optimum reaction conditions may vary with the specific reaction substrate or solvent used, but the conditions can be determined by conventional optimization by those skilled in the art.

[0106] The compounds provided by the present invention can be prepared by standard synthetic methods known in the art. This specification provides a general method for preparing the compounds of the present invention. The starting materials or reagents can usually be obtained commercially, for example, by Alfa It can be purchased from companies such as TCI, or prepared by methods well known to those skilled in the art.

[0107] The compounds of the present invention and corresponding preparation methods are further explained and listed below by examples and preparations. It should be understood that although typical or preferred reaction conditions (such as reaction temperature, time, molar ratio of reactants, reaction solvent, etc.) are given in the specific examples, those skilled in the art may also use other reaction conditions. Optimum reaction conditions may vary with the specific reaction substrate or solvent used, but the conditions may be determined by conventional optimization by those skilled in the art.

[0108] The structures of the following example compounds were characterized by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). 1H-NMR analyses were performed on a Bruker Ascend 400 MHz NMR spectrometer at ambient temperature using TMS as the internal standard and the compounds dissolved in the appropriate deuterated reagent. NMR chemical shifts (δ) are reported in ppm and are abbreviated as follows: s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; brs, broad singlet.

[0109] The reaction starting materials, intermediates and example compounds can be separated and purified by precipitation, filtration, crystallization, evaporation, distillation and chromatography (eg, silica gel column chromatography, preparative chromatography, etc.).

[0110] Preparation of Intermediate 1

[0111] Step 1

[0112] To a 3L three-necked flask, add acetic acid (928 mL) and acetic anhydride (538.0 g) and cool the system to 0°C in an ice bath. Slowly add concentrated sulfuric acid (672.5 g) dropwise to the system, maintaining the system temperature above 40°C. After the additions are complete, cool the system to room temperature again. Then, add 2,3,4-trifluorobenzoic acid (115.0 g), iodine (66.9 g), and manganese dioxide (68.7 g). After the additions are complete, heat the system to 50°C and react for 3 hours. Stop heating, allow the system to cool to room temperature, then add additional iodine (66.9 g) and manganese dioxide (68.7 g). Then, heat the system to 50°C and react for 3 hours. Stop heating, allow the system to cool to room temperature, then add additional iodine (33.4 g) and manganese dioxide (34.4 g). Then, heat the system to 50°C and react for 21 hours. The reaction was stopped, and after the system was cooled to room temperature, it was slowly poured into 11.6L ice water to quench the reaction. Dichloromethane (1500mL) was used for extraction, and the organic phases were combined, washed once with saturated brine (2000mL), and the organic phase was dried over anhydrous sodium sulfate. The organic phase was concentrated (to a volume of about 50mL), and then 1160mL of n-heptane was added to the system and stirred under an ice bath for 30min, during which a large amount of solid was precipitated in the system. Suction filtration was carried out, and the filter cake was washed with 200mL of n-heptane to obtain 178.1g of white solid. ESI-MS: [MH] - =300.9.

[0113] Step 2

[0114] To a 3L three-necked flask, 2,3,4-trifluoro-5-iodobenzoic acid (89.0g), potassium vinyl trifluoroborate (47.4g), potassium phosphate (93.8g), methanol (1780mL), water (890mL), and 1,1'-bis(diphenylphosphino)ferrocenepalladium dichloride (21.6g) were added in sequence. The system was replaced with a nitrogen atmosphere and heated to 60°C for 3h. After cooling to room temperature, the system was filtered through celite, the filtrate was collected, concentrated, and 1M HCl was added to adjust the pH to 4-5. The system was then extracted with methyl tert-ether (500mL). The organic phases were combined, washed with 1L of saturated brine, dried, and concentrated. 750mL of n-heptane and 150mL of dichloromethane were added to the concentrate, stirred at room temperature for 2h, filtered, and the filter cake was washed with 50mL of n-heptane to obtain 48.2g of a white solid. 1 H NMR (400MHz, DMSO-d6): δ7.93-7.87(m,1H),6.80(dd,J1=17.6Hz,J2=11.2Hz,1H),6.01(d,J=18.0Hz,1H),5.57(d,J=10.4Hz,1H).ESI-MS:[MH] - =403.1.

[0115] Step 3

[0116] To a 3L three-necked flask, add 2,3,4-trifluoro-5-vinylbenzoic acid (65.0g), 2-fluoro-4-iodoaniline (76.3g), and tetrahydrofuran (1300mL). The atmosphere was then replaced with nitrogen and the temperature was lowered to -70-80°C. Lithium bistrimethylsilylamide (1M, 965mL) was slowly added dropwise, ensuring the temperature did not exceed -70°C. After the addition was complete, the system was slowly returned to room temperature. 1700mL of hydrochloric acid (1M) was then added to adjust the pH to 1-2. The solution was extracted with methyl tert-ether (500mL). The combined organic phases were washed with 1L of saturated brine, dried, and concentrated to approximately 200mL. 800mL of n-heptane was slowly added to the concentrate, stirred at room temperature for 30min, and filtered to yield 99.0g of an off-white solid. 1 H NMR (400MHz, DMSO-d6): δ9.16 (s, 1H), 7.99-7.97 (m, 1H), 7.64 (dd, J1=10.8Hz, J2=2.0Hz, 1H), 7.43 (dd, J1=8.4Hz, J2=0.8Hz ,1H),6.92-6.84(m,1H),6.78(dd,J1=18.0Hz,J2=11.6Hz,1H),5.91(d,J=17.6Hz,1H),5.47(d,J=11.6Hz,1H).ESI-MS:[MH] - =418.0.

[0117] Step 4

[0118] In a 3L three-necked flask, add 3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-5-vinylbenzoic acid (99.0g), tetrahydrofuran (1980mL) and water (495mL) and stir at room temperature until dissolved. Add osmium tetroxide (3.0g) and sodium periodate (252.7g) to the system and react at room temperature for 1h. Post-treatment: Filter through diatomaceous earth, collect the filtrate, extract with ethyl acetate (500mL), combine the organic phases, wash twice with saturated brine (1000mL), and concentrate the organic phase to obtain a solid crude product. Wash the crude product with anhydrous ethanol (400mL), filter, and wash the filter cake with 200mL of ethanol to obtain 52.4g of off-white solid. 1H NMR (400MHz, DMSO-d6): δ10.22(brs,1H),9.99(s,1H),8.26(d,J=10.8Hz,1H),7.70(dd,J1=1 0.4Hz,J2=1.2Hz,1H),7.52(d,J=8.4Hz,1H),7.10(td,J1=8.4Hz,J2=3.6Hz,1H).ESI-MS:[MH] - =419.9.

[0119] Step 5

[0120] Add 3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-5-formylbenzoic acid (52.4 g) and N,N-dimethylformamide (524 mL) to a 1L three-necked flask and stir the system at room temperature until it becomes clear. Add potassium carbonate (18.9 g) and iodomethane (19.4 g) to the system, then heat the system to 50°C and react for 1 hour. Filter to remove suspended matter in the system and collect the filtrate. Slowly add the filtrate dropwise to water (2100 mL) to precipitate a solid. Continue stirring the system for 1 hour after the addition. Filter and wash the filter cake with 250 mL of ethanol to obtain 50.7 g of a light yellow solid. 1 H NMR (400MHz, DMSO-d6): δ10.00(s,1H),9.52(s,1H),8.21(dd,J1=7.2Hz,J2=1.2Hz,1H),7.71(dd,J1=10 .4Hz,J2=1.6Hz,1H),7.54-7.51(m,1H),7.10(td,J1=8.8Hz,J2=3.2Hz,1H),3.86(s,3H).ESI-MS:[M+H] + =436.0.

[0121] Step 6

[0122] To a 2L three-necked flask, add methyl 3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-5-formylbenzoate (50.7 g), p-methylbenzenesulfonylhydrazide (21.7 g), and ethanol (1500 mL). The system was heated to 50°C and reacted for 5 hours. The system was placed in an ice bath and cooled to 0°C. A large amount of solid precipitated, which was filtered and the filter cake was washed with 100 mL of ethanol to obtain 53.1 g of a light yellow solid. 1H NMR (400MHz, DMSO-d6): δ11.76(s,1H),8.97(s,1H),8.00-7.99(m,2H),7.75(d,J=8.0Hz,2H),7.64(dd,J1=10.4Hz ,J2=1.6Hz,1H),7.43(d,J=8.4Hz,3H),6.89(td,J1=8.8Hz,J2=4.0Hz,1H),3.85(s,3H),2.37(s,3H).ESI-MS:[M+H] + =604.0.

[0123] Step 7

[0124] In a 1L single-necked flask, add 2,3-difluoro-4-iodopyridine (45.0g), (2,4-dimethoxyphenyl)methylamine (78.0g), N-methylpyrrolidone (675mL), and triethylamine (56.7g), and replace the system with a nitrogen atmosphere. Heat the system to 100°C and react for 2h. After the system cools to room temperature, pour it into 700mL of water and extract with tert-methyl ether (500mL). The organic phases are combined, washed with saturated brine, dried, and concentrated to obtain a crude product. The crude product is washed with tert-methyl ether / petroleum ether (1 / 4) to obtain 56.8g of an off-white solid. 1 H NMR (400MHz, DMSO-d6): δ7.45(d,J=5.2Hz,1H),7.10(dd,J1=6.0Hz,J2=5.2Hz,1H),7.03(d,J=8.4Hz,1H),6.88(dd,J1=5.2H z, J2=4.0Hz, 1H), 6.54 (d, J=2.4Hz, 1H), 6.43 (dd, J1=8.4Hz, J2=2.4Hz, 1H), 4.44 (d, J=6.0Hz, 2H), 3.80 (s, 3H), 3.72 (s, 3H).

[0125] Step 8

[0126] In a 2L three-necked flask, add N-(2,4-dimethoxybenzyl)-3-fluoro-4-iodopyridin-2-amine (56.5 g), pinacol diboron (147.8 g), potassium pivalate (44.9 g), toluene (1400 mL), dimethyl sulfoxide (20 mL), and 1,1'-bis(diphenylphosphino)ferrocenepalladium dichloride (10.6 g). The system is then replaced with a nitrogen atmosphere. The system is then heated to 55°C and reacted for 15 hours. After cooling to room temperature, the system is poured into 1000 mL of water and extracted with ethyl acetate (1000 mL). The organic phases are combined, dried over anhydrous sodium sulfate, and concentrated. The concentrate is then diluted with n-hexane (1500 mL), dried, filtered through celite, and the filtrate is concentrated to obtain the product. The product from this step is carried on to the next step without further purification. 1 H NMR (400 MHz, DMSO-d 6 ): δ7.75(dd,J1=4.8Hz,J2=0.8Hz,1H),7.01(d,J=8.4Hz,1H),6.86(td,J1=6.0Hz,J2=4.8Hz,1H),6.57(dd,J1=4.8Hz,J2=3.6Hz,1H),6 .54(d,J=2.4Hz,1H),6.42(dd,J1=8.4Hz,J2=2.4Hz,1H),4.45(d,J=6.0Hz,2H),3.80(s,3H),3.72(s,3H),1.30(s,12H).ESI-MS:[M+H] + =307.1.

[0127] Step 9

[0128] In a 2L single-necked bottle, add N-(2,4-dimethoxybenzyl)-3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-amine and THF (500mL) and stir until dissolved. Slowly add 1M hydrochloric acid aqueous solution (250mL) to the system and stir the reaction at room temperature for 1h. Extract the system with petroleum ether (1000mL) and collect the aqueous phase. Add methylboric acid (17.4g) to the aqueous phase and stir the reaction at room temperature for 1h. Then extract the system with ethyl acetate / petroleum ether (3 / 1) mixed solution (300mL) and collect the aqueous phase. Add ethyl acetate (300mL) to the aqueous phase and adjust the pH of the system to 8 with sodium bicarbonate aqueous solution. Collect the organic phase, wash with water (200mL), dry and concentrate to obtain 37.0g of off-white solid, 1 H NMR (400 MHz, DMSO-d 6): δ8.49(s,2H),7.69(d,J=4.4Hz,1H),7.02(d,J=8.0Hz,1H),6.73-6.70(m,1H),6.58-6.50 (m, 2H), 6.42 (dd, J1 = 8.4Hz, J2 = 2.4Hz, 1H), 4.44 (d, J = 6.0Hz, 2H), 3.81 (s, 3H), 3.72 (s, 3H).

[0129] Step 10

[0130] To a 1L single-necked flask, (2-((2,4-dimethoxybenzyl)amino)-3-fluoropyridin-4-yl)boric acid (15.0 g), methyl (E)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-5-((2-toluenesulfonylhydrazide)methyl)benzoate (35.4 g), potassium carbonate (8.1 g), and toluene (600 mL) were added, and the system was replaced with a nitrogen atmosphere. The system was heated to 100°C and reacted for 5 h. After the system was cooled to room temperature, saturated aqueous ammonium chloride (300 mL) was added, and the mixture was extracted with ethyl acetate (100 mL). The organic phases were combined and dried over anhydrous sodium sulfate. The solvent was removed by filtration, and the crude product was then purified by 100-200 mesh silica gel column chromatography to obtain 17.9 g of a light yellow liquid. 1 H NMR (400 MHz, DMSO-d 6 ): δ8.65(s,1H),7.73(d,J=7.2Hz,1H),7.68(d,J=5.2Hz,1H),7.62(dd,J1=10.8Hz,J2=3.0 Hz,1H),7.39(d,J=8.4Hz,1H),7.03(d,J=8.4Hz,1H),6.92-6.88(m,1H),6.78(td,J1=8.8Hz ,J2=4.4Hz,1H),6.54(d,J=2.4Hz,1H),6.42(dd,J1=8.4Hz,J2=4.4Hz,1H),6.38(t,J=4.8Hz ,1H),4.44(d,J=6.0Hz,2H),4.01(s,2H),3.80(d,J=1.6Hz,6H),3.72(s,3H).ESI-MS:[M+H] + =682.1.

[0131] Step 11

[0132] In a 500 mL single-necked bottle, methyl 5-((2-((2,4-dimethoxybenzyl)amino)-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoate (10.0 g) and dichloromethane (150 mL) were added and stirred at room temperature until dissolved. The system was then placed in an ice bath and trifluoroacetic acid (150 mL) was slowly added. After the addition was complete, the system was warmed to room temperature and stirred for 1.5 hours. The system was slowly added to a saturated aqueous sodium carbonate solution and the pH of the system was adjusted to 8. Extracted with dichloromethane (100 mL), the organic phases were combined and dried over anhydrous sodium sulfate. Filtered and the solvent was removed by rotation, and the crude product was then purified by 100-200 mesh silica gel column chromatography to obtain 5.8 g of a light yellow solid. 1 H NMR (400MHz, DMSO-d6): δ8.64(s,1H),7.71(d,J=7.2Hz,1H),7.67(d,J=5.2Hz,1H),7.62(dd,J1=10.8Hz,J2=2.4Hz,1H),7.39(dd,J1=8.8Hz,J 2=1.2Hz,1H),6.77(td,J1=8.8Hz,J2=4.8Hz,1H),6.40(t,J=4.8Hz,1H),6.22(brs,2H),3.99(s,2H),3.79(s,3H),3.08(s,2H).ESI-MS:[M+H] + =532.0.

[0133] Step 12

[0134] In a 1L single-necked flask, methyl 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoate (6.9 g), tetrahydrofuran (100 mL) and water (50 mL) were added in sequence, and the system was placed in an ice bath and stirred. Lithium hydroxide monohydrate (1.6 g) was slowly added to the system. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hours. Aqueous hydrochloric acid solution (6 M, 20 mL) was then added to the system to adjust the pH to 1-2 and stirred for 5 minutes. The tetrahydrofuran in the system was removed by concentration under reduced pressure, and then 150 mL of water was added to the system to dilute the concentrate and stir for 10 minutes. The filter cake was collected by filtration and washed once with water (50 mL) and tert-methyl ether (50 mL) to obtain 6.2 g of a purple-red solid product. ESI-MS: [M+H] + =518.0.

[0135] Preparation of Intermediate 2

[0136] Step 1:

[0137] In a 2L three-necked flask, 2,3,4-trifluoro-5-vinylbenzoic acid (24.2g), 2-fluoro-4-methylaniline (15.0g) and tetrahydrofuran (485mL) were added, and then the system was replaced with a nitrogen environment and cooled to -70~-80°C. Lithium bis(trimethylsilyl)amide (1M, 360mL) was slowly added dropwise to the system, ensuring that the system temperature did not exceed -70°C. After the dropwise addition, the system was slowly restored to room temperature, and 600mL of hydrochloric acid (1M) was added to adjust the pH to 1~2. Extracted with tert-methyl ether, the organic phases were combined, washed with saturated brine, dried and concentrated. Slowly add n-heptane to the concentrate, stir at room temperature for 30min, filter, and obtain 30.4g of light yellow solid. ESI-MS: [MH] - =306.1.

[0138] Step 2:

[0139] In a 2L three-necked flask, water (100mL), tetrahydrofuran (300mL) and acetonitrile (300mL) were added and stirred at room temperature. Subsequently, concentrated sulfuric acid (2.0g) was slowly added dropwise to the reaction flask. After the system temperature returned to room temperature, sodium periodate (34.9g) and anhydrous ruthenium trichloride (0.5g) were slowly added thereto and the reaction was continued to stir at room temperature. After 5min, 3,4-difluoro-2-((2-fluoro-4-methylphenyl)amino)-5-vinylbenzoic acid (10.0g) was added to the system and the reaction was stirred at room temperature for 2h. The insoluble matter was removed by filtration, the filtrate was collected, extracted with ethyl acetate, the organic phases were combined, washed with saturated sodium sulfite aqueous solution, then washed with saturated brine, the organic phases were collected and dried, filtered, and concentrated to obtain 7.0g of a yellow solid. ESI-MS: [MH] - =308.1.

[0140] Step 3:

[0141] 3,4-difluoro-2-((2-fluoro-4-methylphenyl)amino)-5-formylbenzoic acid (3.1 g) and N,N-dimethylformamide (31 mL) were added to a 250 mL three-necked flask and stirred at room temperature until the mixture became clear. Potassium carbonate (1.5 g) and iodomethane (1.6 g) were added to the system, and the temperature was then raised to 50°C for 4 h. The suspended solids were removed by filtration, and the filtrate was collected. The filtrate was slowly added dropwise to water to precipitate a solid. Stirring was continued for 1 h after the addition was complete. The filter cake was washed with water and dried to obtain 1.3 g of a yellow solid. ESI-MS: [M+H] + =324.1.

[0142] Step 4:

[0143] To a 2L three-necked flask, add methyl 3,4-difluoro-2-((2-fluoro-4-methylphenyl)amino)-5-formylbenzoate (1.8 g), p-methylbenzenesulfonylhydrazide (1.1 g), and ethanol (54 mL). The system was heated to 50°C and reacted for 2 h. The system was placed in an ice bath and cooled to 0°C. A large amount of solid precipitated, which was filtered and the filter cake washed with ethanol to obtain 1.7 g of a light yellow solid. 1 H NMR (400MHz, DMSO-d6): δ11.69(s,1H),9.04(s,1H),8.01-7.98(m,2H),7.75(d,J=8.0Hz,2H),7.43(d,J=8 .0Hz,2H),7.09-7.03(m,2H),6.93(d,J=8.0Hz,1H),3.87(s,3H),2.38(s,3H),2.28(s,3H).ESI-MS:[M+H] + =492.1.

[0144] Step 5:

[0145] In a 1L single-necked flask, (2-((2,4-dimethoxybenzyl)amino)-3-fluoropyridin-4-yl)boric acid (2.7 g), (E)-3,4-difluoro-2-((2-fluoro-4-methylphenyl)amino)-5-((2-toluenesulfonylhydrazono)methyl)benzoic acid methyl ester (3.6 g), potassium carbonate (1.2 g) and toluene (144 mL) were added, and the system was replaced with a nitrogen atmosphere. The system was heated to 100°C and reacted for 4 hours. After the system was cooled to room temperature, saturated aqueous ammonium chloride solution was added, extracted with ethyl acetate, and the organic phases were combined and dried over anhydrous sodium sulfate. The solvent was removed by filtration, and the crude product was then purified by silica gel column chromatography to obtain 3.2 g of a light yellow liquid. ESI-MS: [M+H] + =570.2.

[0146] Step 6:

[0147] In a 250mL single-necked bottle, 5-((2-((2,4-dimethoxybenzyl)amino)-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-methylphenyl)amino)benzoic acid methyl ester (3.2g) and dichloromethane (60mL) were added and stirred at room temperature until clear. The system was then placed in an ice bath and trifluoroacetic acid (60mL) was slowly added. After the addition was completed, the system was warmed to room temperature and stirred for 1.5h. Saturated aqueous sodium carbonate solution was slowly added to the system to adjust the pH to 8. Extracted with dichloromethane, the organic phases were combined and dried over anhydrous sodium sulfate. Filtered and the solvent was spun off, and the crude product was then purified by silica gel column chromatography to obtain 2.1g of a light yellow solid. ESI-MS: [M+H] + =420.1.

[0148] Step 7:

[0149] In a 250 mL single-necked bottle, methyl 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-methylphenyl)amino)benzoate (2.5 g), tetrahydrofuran (60 mL) and water (30 mL) were added in sequence, and the system was placed in an ice bath and stirred. Lithium hydroxide monohydrate (0.8 g) was slowly added to the system. After the addition was complete, the mixture was warmed to room temperature and stirred for 4 hours. Aqueous hydrochloric acid solution (6 M, 10 mL) was then added to the system to adjust the pH to 1-2 and stirred for 5 minutes. The tetrahydrofuran in the system was removed by concentration under reduced pressure, and then the concentrate was diluted with water and stirred for 10 minutes. Filter, collect the filter cake, and wash it once with water and tertiary methyl ether respectively to obtain 1.4 g of purple-red solid. ESI-MS: [M+H] + =406.1.

[0150] Preparation of Intermediate 3

[0151] Step 1:

[0152] In a 2L three-necked flask, add 2,3,4-trifluoro-5-vinylbenzoic acid (20.0g), aniline (9.2g) and tetrahydrofuran (400mL), then replace the system with a nitrogen atmosphere and cool to -15~-20℃. Slowly add lithium bis(trimethylsilyl)amide (1M, 297mL) dropwise to the system, ensuring that the system temperature does not exceed -15℃. After the addition is complete, slowly return the system to room temperature, then add hydrochloric acid (6M) to the system to adjust the pH of the system to 1~2. Use ethyl acetate to extract the system, combine the organic phases, wash with 1L of saturated brine, dry and concentrate to obtain 24.5g of yellow solid. 1 H NMR (400MHz, DMSO-d6): δ13.69(s,1H),9.13(s,1H),7.97(d,J=7.2Hz,1H),7.27(t,J=8.0Hz,2H),7.00-6.96 (m,3H),6.78(dd,J1=17.6Hz,J2=11.2Hz,1H),5.89(d,J=18.0Hz,1H),5.44(d,J=11.6Hz,1H).ESI-MS:[M+H] + =276.1.

[0153] Step 2:

[0154] 3,4-difluoro-2-(phenylamino)-5-vinylbenzoic acid (30.0 g) and N,N-dimethylformamide (300 mL) were added to a 1 L three-necked flask and stirred at room temperature until the solution became clear. Potassium carbonate (22.6 g) and iodomethane (20.1 g) were added to the system, and the system was then reacted at room temperature for 16 h. The suspended matter in the system was filtered out, and the filtrate was collected. The filtrate was slowly added dropwise to water to precipitate a solid. After the addition was complete, the system was stirred for another 1 h. Filtered with suction, and the filter cake was washed with ethanol to obtain 21.0 g of a light yellow solid. 1 H NMR (400MHz, DMSO-d6): δ8.77(s,1H),7.91(d,J=6.8Hz,1H),7.25(t,J=8.0Hz,2H),6.97-6.96(m,3H),6.78 (dd, J1=17.6Hz, J2=11.2Hz, 1H), 5.91 (d, J=17.6Hz, 1H), 5.46 (d, J=11.6Hz, 1H), 3.78 (s, 3H).ESI-MS: [M+H] + =290.1.

[0155] Step 3:

[0156] In a 1L three-necked flask, methyl 3,4-difluoro-2-(phenylamino)-5-vinylbenzoate (20.8g) and tetrahydrofuran (312mL) were added and stirred in an ice bath until dissolved. Potassium osmate dihydrate (1.3g) was added to the system and the reaction was continued by stirring in an ice bath. After 15min, water (312mL), sodium periodate (46.2g) and 2,6-lutidine (7.7g) were added to the system, and the system was heated to room temperature and stirred for 4h. The reaction system was poured into a saturated aqueous sodium sulfite solution, extracted with ethyl acetate, the organic phases were combined, washed twice with saturated brine, and the organic phase was concentrated to obtain 20.0g of a yellow solid. ESI-MS: [M+H] + =292.1.

[0157] Step 4:

[0158] Methyl 3,4-difluoro-2-(phenylamino)-5-formylbenzoate (20.0 g), p-methylbenzenesulfonylhydrazide (12.8 g), and ethanol (600 mL) were added to a 2 L three-necked flask and the reaction was heated to 50°C for 15 h. The system was cooled to 0°C in an ice bath to precipitate a large amount of solid. The solid was filtered and the filter cake was washed with 50 mL of ethanol to obtain 26.0 g of an off-white solid. 1H NMR (400MHz, DMSO-d6): δ11.71(s,1H),8.98(s,1H),8.00-7.97(m,2H),7.75(d,J=8.0Hz,2H),7.43 (d,J=8.0Hz,2H),7.26(t,J=8.0Hz,2H),6.99-6.98(m,3H),3.80(s,3H),2.38(s,3H).ESI-MS:[M+H] + =460.1.

[0159] Step 5:

[0160] In a 1L single-necked flask, (2-((2,4-dimethoxybenzyl)amino)-3-fluoropyridin-4-yl)boric acid (6.9 g), (E)-3,4-difluoro-2-(phenylamino)-5-((2-toluenesulfonylhydrazono)methyl)benzoate (8.0 g), potassium carbonate (3.6 g) and toluene (320 mL) were added, and the system was replaced with a nitrogen atmosphere. The system was heated to 100°C and reacted for 5 h. After the system was cooled to room temperature, saturated aqueous ammonium chloride was added, extracted with ethyl acetate, and the organic phases were combined and dried over anhydrous sodium sulfate. The solvent was removed by filtration to obtain 4.7 g of a light yellow liquid. ESI-MS: [M+H] + =538.2.

[0161] Step 6:

[0162] In a 500mL single-necked bottle, methyl 5-((2-((2,4-dimethoxybenzyl)amino)-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-(phenylamino)benzoate (4.7g) and dichloromethane (220mL) were added and stirred at room temperature until dissolved. The system was then placed in an ice bath and trifluoroacetic acid (110mL) was slowly added. After the addition was completed, the system was warmed to room temperature and stirred for 1.5h. The system was slowly added to a saturated aqueous sodium carbonate solution and the pH of the system was adjusted to 8. Extracted with dichloromethane, the organic phases were combined and dried over anhydrous sodium sulfate. Filtered and the solvent was spun off, and the crude product was then purified by silica gel column chromatography to obtain 1.8g of a yellow solid.

[0163] Step 7:

[0164] In a 500 mL single-necked flask, methyl 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-(phenylamino)benzoate (1.8 g), tetrahydrofuran (40 mL), and water (40 mL) were added sequentially and the system was stirred in an ice bath. Lithium hydroxide monohydrate (586 mg) was slowly added to the system. After addition, the mixture was warmed to room temperature and stirred for 2 h. Aqueous hydrochloric acid (6 M, 20 mL) was then added to the system to adjust the pH to 1-2 and stirred for 5 min. The tetrahydrofuran in the system was removed by concentration under reduced pressure, and then the concentrate was diluted with water and stirred for 10 min. The filter cake was collected by filtration and washed with an acetone / n-hexane mixed solvent to obtain 1.4 g of a light yellow solid.

[0165] Preparation of Intermediate 4

[0166] Step 1:

[0167] In a 1L three-necked flask, 2,3,4-trifluoro-5-vinylbenzoic acid (10.0g), 2-fluoro-4-methoxyaniline (7.0g) and tetrahydrofuran (250mL) were added, and then the system was replaced with a nitrogen environment and cooled to -15 to -20°C. Lithium bis(trimethylsilyl)amide (1M, 150mL) was slowly added dropwise to the system, ensuring that the system temperature did not exceed -15°C. After the dropwise addition, the system was slowly restored to room temperature, and then sodium bisulfate aqueous solution (1.5M) was added to the system to adjust the pH of the system to 1 to 2. The system was extracted with tert-methyl ether, the organic phases were combined, washed with 1L saturated brine, dried and concentrated to obtain 15.1g of a yellow solid. ESI-MS: [MH] - =322.1.

[0168] Step 2:

[0169] 3,4-difluoro-2-(2-fluoro-4-methoxyphenylamino)-5-vinylbenzoic acid (16.2 g) and N,N-dimethylformamide (160 mL) were added to a 1L three-necked flask and the system was stirred at room temperature until dissolved. Potassium carbonate (8.3 g) and iodomethane (8.5 g) were added to the system, and the system was then reacted at 35°C for 4 h. The suspended matter in the system was removed by filtration, and the filtrate was collected. The filtrate was slowly added dropwise to water to precipitate a solid. After the addition was complete, the system was stirred for 1 h. Filtered and the filter cake was washed with n-heptane to obtain 16.1 g of a light yellow solid. ESI-MS: [M+H] + =338.1.

[0170] Step 3:

[0171] In a 500ml three-necked flask, add 3,4-difluoro-2-(2-fluoro-4-methoxyphenylamino)-5-vinylbenzoic acid methyl ester (15.8g) and tetrahydrofuran (314mL) and stir in an ice bath until dissolved. Add potassium osmate dihydrate (0.86g) to the system and continue stirring the reaction in an ice bath. After 15min, add water (312mL), sodium periodate (30.0g) and 2,6-lutidine (5.1g) to the system, and warm the system to room temperature and stir the reaction for 4h. Pour the reaction system into a saturated aqueous sodium sulfite solution, extract with ethyl acetate, combine the organic phases, wash with saturated brine, and concentrate. Dilute the concentrate with acetone, slowly add water to the system, stir, filter, and dry the filter cake to obtain 12.7g of light yellow solid.

[0172] Step 4:

[0173] To a 500 mL three-necked flask, add methyl 3,4-difluoro-2-((2-fluoro-4-methoxyphenyl)amino)-5-formylbenzoate (4.0 g), p-methylbenzenesulfonylhydrazide (2.2 g), and ethanol (120 mL). The system was heated to 60°C and reacted for 1 h. The system was placed in an ice bath and cooled to 0°C. A large amount of solid precipitated, which was filtered and the filter cake washed with ethanol to obtain 5.1 g of a light yellow solid. ESI-MS: [M+H] + =508.1.

[0174] Step 5:

[0175] In a 500 mL single-necked flask, (2-((2,4-dimethoxybenzyl)amino)-3-fluoropyridin-4-yl)boric acid (3.6 g), (E)-3,4-difluoro-2-((2-fluoro-4-methoxyphenyl)amino)-5-((2-toluenesulfonylhydrazono)methyl)benzoate (5.0 g), potassium carbonate (1.63 g), and toluene (150 mL) were added, and the system was replaced with a nitrogen atmosphere. The system was heated to 105°C and reacted for 3 hours. After the system was cooled to room temperature, saturated aqueous ammonium chloride was added, extracted with ethyl acetate, and the organic phases were combined and dried over anhydrous sodium sulfate. The solvent was removed by filtration, and the crude product was purified by silica gel column chromatography to obtain 4.4 g of a light yellow liquid. ESI-MS: [M+H] + =586.2.

[0176] Step 6:

[0177] In a 500mL single-necked bottle, 5-((2-((2,4-dimethoxybenzyl)amino)-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-methoxyphenyl)amino)benzoic acid methyl ester (4.4g) and dichloromethane (27mL) were added and stirred at room temperature until dissolved. The system was then placed in an ice bath and trifluoroacetic acid (9.1mL) was slowly added. After the addition was completed, the system was warmed to room temperature and stirred for 1.5h. The system was slowly added to a saturated aqueous sodium carbonate solution and the pH of the system was adjusted to 8. Extracted with dichloromethane, the organic phases were combined and dried over anhydrous sodium sulfate. Filtered and the solvent was removed by spin to obtain 3.1g of a light yellow solid. ESI-MS: [M+H] + =436.1.

[0178] Step 7:

[0179] In a 50 mL single-necked bottle, methyl 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-methoxyphenyl)amino)benzoate (3.0 g), acetonitrile (18 mL) and ethanol (12 mL) were added in sequence, and the system was placed in an ice bath and stirred. An aqueous solution (18 mL) of lithium hydroxide monohydrate (870 mg) was slowly added dropwise to the system. After the addition was complete, the mixture was heated to room temperature and stirred for 2.5 hours. A 1.5 M aqueous sodium bisulfate solution was then added to the system to adjust the pH to 1-2 and stirred for 5 minutes. The filter cake was collected by filtration and washed once with water and tertiary methyl ether to obtain 2.7 g of an off-white solid. ESI-MS: [M+H] + =422.1.

[0180] Preparation of Intermediate 5

[0181] Step 1:

[0182] In a 250 mL single-necked flask, add 3-bromo-2-fluoroaniline (10.1 g), 2,3,4-trimethoxybenzaldehyde (10.0 g), and toluene (200 mL). Heat the mixture to reflux and stir for 2 hours. Remove from heat, cool the mixture to room temperature, concentrate, and dilute the concentrate with 150 mL of 1,2-dichloroethane. Add 3 mL of acetic acid and sodium triacetoxyborohydride (21.0 g) to the mixture and stir for 12 hours at room temperature. Pour the mixture into water and extract with dichloromethane. Combine the organic phases, dry, and concentrate to yield 17.0 g of a brown solid.

[0183] Step 2:

[0184] To a 250 mL single-necked flask, add 3-bromo-2-fluoro-N-(2,3,4-trimethoxybenzyl)aniline (5.2 g), pinacol diboron (7.2 g), potassium pivalate (4.0 g), toluene (125 mL), dimethyl sulfoxide (2.2 g), and [1,1′-bis(diphenylphosphino)ferrocene]palladium dichloride (1.0 g). The system was then replaced with a nitrogen atmosphere. The system was then heated to 55°C and reacted for 19 hours. After cooling to room temperature, the mixture was poured into water and extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. The concentrate was then diluted with n-hexane, dried, and filtered through celite. The filtrate was concentrated to yield 4.7 g of a yellow solid.

[0185] Step 3:

[0186] 2-Fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-N-(2,3,4-trimethoxybenzyl)aniline (4.7 g) and tetrahydrofuran (100 mL) were added to a 250 mL single-necked bottle and stirred until dissolved. 1 M aqueous hydrochloric acid solution (50 mL) was slowly added to the system and stirred at room temperature for 1 hour. Methylboric acid (2.4 g) was added to the system and stirred at room temperature for 24 hours. The system was then extracted with petroleum ether, the organic phases were combined, dried, and concentrated to obtain 1.1 g of a light yellow solid. ESI-MS: [M+H] + =336.1.

[0187] Step 4:

[0188] In a 100 mL single-necked flask, (2-fluoro-3-((2,3,4-trimethoxybenzyl)amino)phenyl)boric acid (1.1 g), (E)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-5-((2-toluenesulfonylhydrazono)methyl)benzoate (2.4 g), potassium carbonate (540 mg), and toluene (440 mL) were added, and the system was replaced with a nitrogen atmosphere. The system was heated to 100°C and reacted for 5 h. After the system was cooled to room temperature, saturated aqueous ammonium chloride was added, extracted with ethyl acetate, and the organic phases were combined and dried over anhydrous sodium sulfate. The solvent was removed by filtration, and the crude product was purified by silica gel column chromatography to obtain 1.1 g of a light yellow liquid. ESI-MS: [M+H] + =711.1.

[0189] Step 5:

[0190] In a 100mL single-necked bottle, methyl 3,4-difluoro-5-(2-fluoro-3-((2,3,4-trimethoxybenzyl)amino)benzyl)-2-((2-fluoro-4-iodophenyl)amino)benzoate (1.9g) and dichloromethane (30mL) were added and stirred at room temperature until dissolved. The system was then placed in an ice bath and trifluoroacetic acid (30mL) was slowly added. After the addition was completed, the system was heated to room temperature and stirred for 3.5h. p-Toluenesulfonic acid (515mg) was added to the system, and the system was heated to 40°C and stirred for 24h. After concentrating the system, the concentrate was diluted with saturated aqueous sodium carbonate solution and its pH was adjusted to 8. Extracted with ethyl acetate, the organic phases were combined, dried, filtered and concentrated. The concentrate was purified by silica gel column chromatography to obtain 670mg of a light yellow solid. ESI-MS: [M+H] + =531.0.

[0191] Step 6:

[0192] In a 100 mL single-necked bottle, methyl 5-(3-amino-2-fluorobenzyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoate (670 mg), tetrahydrofuran (20 mL) and water (10 mL) were added in sequence, and the system was placed in an ice bath and stirred. Lithium hydroxide monohydrate (160 g) was slowly added to the system. After the addition was complete, the mixture was warmed to room temperature and stirred for 13 hours. Aqueous hydrochloric acid solution (2 M) was then added to the system to adjust the pH to 1-2 and stirred for 5 minutes. The tetrahydrofuran in the system was removed by concentration under reduced pressure, and then water was added to the system to dilute the concentrate and stir for 10 minutes. Filter to obtain 630 mg of a light yellow solid. ESI-MS: [M+H] + =517.0.

[0193] Preparation of Intermediate 6

[0194] Under ice bath conditions, 120 mL of acetonitrile and chlorosulfonyl isocyanate (3.2 g) were added to a 500 mL single-necked bottle. 2-bromoethanol (2.8 g) was then slowly added to the system. The system was kept stirring under ice bath for 30 minutes, then heated to room temperature and stirred for 1 hour. N-methylmorpholine (15.1 g) was then slowly added to the system and stirred for 10 minutes, and then ethylamine hydrochloride (1.8 g) was slowly added to the system. After the addition, the system was heated to 50°C and stirred for 5 hours. The system was then gradually cooled to room temperature and continued to stir for 15 hours. Filter and concentrate the filtrate. Ethyl acetate was added to dilute the concentrate, washed with saturated brine, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain 1.8 g of a white solid. 1H NMR (400MHz, DMSO-d6, δ): 8.34 (t, J = 5.2 Hz, 1H), 4.41-4.37 (m, 2H), 3.96-3.92 (m, 2H), 3.08-3.01 (m, 2H), 1.08 (t, J = 7.2 Hz, 3H).

[0195] Preparation of Intermediate 7

[0196] Under ice conditions, add 500 mL of acetonitrile and chlorosulfonyl isocyanate (11.6 g) to a 1 L single-necked flask. Subsequently, slowly add 2-bromoethanol (10.3 g) to the system. The system is stirred and reacted for 30 minutes while still in the ice bath, then warmed to room temperature and stirred for 1 hour. Next, slowly add N-methylmorpholine (50.0 g) to the system and stir for 10 minutes, followed by slowly adding cyclopropylamine (4.7 g) to the system. After addition, warm the system to 50°C and stir for 5 hours. The system is then gradually cooled to room temperature and stirred for another 15 hours. Filter and concentrate the filtrate to obtain a light orange solid. Ethyl acetate is added to dissolve most of the solid, and the mixture is washed with saturated brine. The organic phase is separated and dried over anhydrous sodium sulfate. Filter and remove the solvent to obtain the crude product. The crude product is washed with tert-methyl ether to obtain 4.4 g of an off-white solid. 1 H NMR (400MHz, DMSO-d6, δ): 8.72 (s, 1H), 4.44-4.40 (m, 2H), 4.00-3.96 (m, 2H), 2.50-2.45 (m, 1H), 0.65-0.58 (m, 2H), 0.56-0.52 (m, 2H).

[0197] Preparation of Intermediate 8

[0198] Under ice bath conditions, 720mL of acetonitrile and chlorosulfonyl isocyanate (20.6g) were added to a 2L single-necked flask. 2-bromoethanol (18.2g) was then slowly added to the system. The system was kept stirring under ice bath for 30min, then heated to room temperature and stirred for 1h. N-methylmorpholine (58.4g) was then slowly added to the system and stirred for 10min, after which deuterated methylamine hydrochloride (10.2g) was slowly added to the system. After the addition, the system was heated to 50°C and stirred for 5h. The system was then gradually cooled to room temperature and stirred for another 15h. The filtrate was filtered and concentrated to obtain an orange viscous liquid. The viscous liquid was diluted with water, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, and the organic phases were dried over anhydrous sodium sulfate. Filtered and the solvent was removed by spinning to obtain 11.9g of a light yellow solid. 1H NMR (400MHz, DMSO-d6, δ): 8.18 (s, 1H), 4.42-4.38 (m, 2H), 3.96-3.93 (m, 2H).

[0199] Compound preparation

[0200] Example 1:

[0201] Step 1:

[0202] In a 100 mL single-necked flask, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoic acid hydrochloride (1.7 g), 1-hydroxybenzotriazole (1.0 g), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.3 g), and N,N-dimethylformamide (20 mL) were added and stirred at room temperature for 3.5 hours. Propargylamine (542 mg) and N,N-diisopropylethylamine (1.3 g) were then added, and the reaction was stirred at room temperature for another 2 hours. The reaction system was added to water (200 mL) and extracted with ethyl acetate (100 mL). The organic phase was collected, washed with saturated brine (100 mL), dried, and concentrated. The crude product was then purified by 100-200 mesh silica gel column chromatography to yield 770 mg of an off-white solid. 1H NMR (400MHz, DMSO-d6): δ9.17(t,J=5.2Hz,1H),9.03(s,1H),7.66(d,J=4.8Hz,1 H),7.59(dd,J1=10.8Hz,J2=2.0Hz,1H),7.53(d,J=6.8Hz,1H),7.36(dd,J1=8.4H z,J2=1.2Hz,1H),6.69(td,J1=8.8Hz,J2=4.8Hz,1H),6.40(t,J=4.8Hz,1H),6.1 9(brs,2H),4.02-4.00(m,2H),3.96(s,2H),3.16(t,J=2.8Hz,1H).ESI-MS:[M+H] + =555.0.

[0203] Step 2

[0204] To a 50 mL single-necked flask, add 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-N-(alkyn-2-prop-1-yl)benzamide (770 mg), N,N-dimethylformamide (8 mL), and pyridine (1340 mg), and stir in an ice bath. Once the temperature drops below 0°C, slowly add methylaminosulfonyl chloride dropwise (in four batches, 10 min apart, for a total of 1250 mg, 9.6 mmol). After 30 min, the reaction is terminated, and the system is poured into saturated aqueous sodium bicarbonate (100 mL) and stirred at room temperature for 10 min. Extract with ethyl acetate (50 mL), combine the organic phases, and dry over anhydrous sodium sulfate. Filter and remove the solvent, then purify the crude product using a 100-200 mesh silica gel column to yield 220 mg of an off-white solid. 1 H NMR (400MHz, DMSO-d6): δ10.42(s,1H),9.18(t,J=5.2Hz,1H),9.06(s,1H),8.02(d,J=5.2Hz, 1H),7.59(dd,J1=10.8Hz,J2=2.0Hz,1H),7.56(d,J=6.8Hz,1H),7.37(dd,J1=8.4Hz,J2=1.2H z,1H),7.03(q,J=4.8Hz,1H),6.95(t,J=4.8Hz,1H),6.71(td,J1=8.8Hz,J2=5.2Hz,1H),4.07 (s,2H),4.01(dd,J1=5.2Hz,J2=2.4Hz,2H),3.17(t,J=2.4Hz,1H),2.52(s,3H).ESI-MS:[M+H] + =648.0.

[0205] Example 2

[0206] Step 1:

[0207] In a 100 mL single-necked flask, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoic acid hydrochloride (600 mg), 1-hydroxybenzotriazole (297 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (844 mg), and N,N-dimethylformamide (32 mL) were added and stirred at room temperature for 2 hours. Subsequently, but-3-yn-1-amine (150 mg) and N,N-diisopropylethylamine (711 mg) were added to the system, and the reaction was stirred at room temperature for another 2 hours. The reaction system was added to saturated aqueous sodium bicarbonate solution to precipitate a solid. The filter cake was filtered and washed with ethyl acetate / petroleum ether (1 / 3) to obtain 500 mg of a light yellow solid. 1 H NMR (400MHz, DMSO-d6): δ9.00(s,1H),8.84(t,J=5.6Hz,1H),7.66(d,J=4.8Hz,1 H),7.58(dd,J1=10.8Hz,J2=2.0Hz,1H),7.50(d,J=6.4Hz,1H),7.36(dd,J1=8.8H z,J2=0.8Hz,1H),6.70-6.64(m,1H),6.39(t,J=4.8Hz,1H),6.17(s,2H),3.96(s ,2H),3.32-3.28(m,2H),2.79(t,J=2.4Hz,1H),2.37-2.33(m,2H).ESI-MS:[M+H] + =569.1.

[0208] Step 2:

[0209] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-N-(but-3-yn-1-yl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzamide (520 mg), N,N-dimethylformamide (7.5 mL) and pyridine (724 mg) were added, and the system was placed in an ice bath and stirred. After the system cooled to below 0°C, methylaminosulfonyl chloride (1.3 g) was slowly added dropwise. After 30 minutes, the reaction was stopped, and the system was poured into a saturated aqueous sodium bicarbonate solution and stirred at room temperature for 10 minutes. Extracted with ethyl acetate, the organic phases were combined and dried over anhydrous sodium sulfate. Filtered and the solvent was removed by rotation, and the crude product was purified by silica gel column chromatography to obtain 220 mg of an off-white solid. 1H NMR (400MHz, DMSO-d6): δ10.35(s,1H),9.03(s,1H),8.84(s,1H),8.02(d,J=3.6Hz,1H),7.58(d,J=10.8Hz,1H),7.52(d,J=6.4Hz,1H),7.37(d,J=8.0 Hz,1H),7.02(d,J=3.6Hz,1H),6.93(s,1H),6.72-6.69(m,1H),4.07(s,2H),3.3 1-3.30(m,2H),2.79(brs,1H),2.53-2.52(m,3H),2.36(brs,2H).ESI-MS:[M+H] + =662.0.

[0210] Example 3

[0211] To a 100 mL single-necked flask, add N-methyl-d3-2-oxoxazoline-3-sulfonamide (1.3 g), 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-N-(alkyn-2-prop-1-yl)benzamide (400 mg), triethylamine (804 mg), and 1,2-dichloroethane (40 mL). The reaction system was heated to 80°C for 3.5 hours. After cooling to room temperature, the system was poured into a water / tetrahydrofuran mixture (90 / 10, v / v). The aqueous phase was extracted with ethyl acetate, and the organic phases were combined, dried, and concentrated. The crude product was purified by silica gel column chromatography to obtain the crude product. The crude product was washed with a n-hexane / acetone mixture (4 / 1, v / v) to obtain 54 mg of an off-white solid. 1 H NMR (400MHz, DMSO-d6): δ10.42(s,1H),9.18(t,J=5.6Hz,1H),9.06(s,1H),8.02(d,J=4.8Hz,1H),7.59(dd,J1=10.8Hz,J2=1.6Hz,1H),7.56(d,J=6.8Hz ,1H),7.37(d,J=8.4Hz,1H),7.01(s,1H),6.94(t,J=4.4Hz,1H),6.74-6.68( m,1H),4.07(s,2H),4.02-4.00(m,2H),3.17(t,J=2.4Hz,1H).ESI-MS:[M+H] + =651.1.

[0212] Example 4

[0213] Step 1:

[0214] To a 100 mL single-necked flask, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoic acid hydrochloride (600 mg), 1-hydroxybenzotriazole (297 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (844 mg), and N,N-dimethylformamide (32 mL) were added and stirred at room temperature for 2 h. (S)-but-3-yn-2-amine hydrochloride (229 mg) and N,N-diisopropylethylamine (711 mg) were then added and stirred at room temperature for another 2 h. The reaction system was added to saturated aqueous sodium bicarbonate solution (100 mL) to precipitate a solid. The filter cake was filtered and washed with ethyl acetate / petroleum ether (1 / 3) to yield 500 mg of a light yellow solid. 1 H NMR (400MHz, DMSO-d6): δ9.07(d,J=7.6Hz,1H),8.91(s,1H),7.66(d,J=5.2Hz,1H),7.58(dd,J1=10.8Hz,J2 =2.0Hz,1H),7.54(d,J=6.8Hz,1H),7.35(dd,J1=8.4Hz,J2=1.2Hz,1H),6.69-6.63(m,1H),6.38(t,J=4.8Hz 1H),6.17(s,2H),4.78-4.70(m,1H),3.97(s,2H),3.22(d,J=2.0Hz,1H),1.33(d,J=7.2Hz,3H).ESI-MS:[M+H] + =569.0.

[0215] Step 2:

[0216] In a 50 mL single-necked flask, (R)-5-((2-amino-3-fluoropyridin-4-yl)methyl)-N-(but-3-yn-2-yl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzamide (500 mg), N,N-dimethylformamide (10 mL) and pyridine (595 mg) were added, and the system was placed in an ice bath and stirred. After the system cooled to below 0°C, methylaminosulfonyl chloride (700 mg) was slowly added dropwise. The reaction was stopped after 30 minutes, and the system was poured into a saturated aqueous sodium bicarbonate solution and stirred at room temperature for 10 minutes. Extracted with ethyl acetate, the organic phases were combined and dried over anhydrous sodium sulfate. Filtered and the solvent was removed by rotation, and the crude product was purified by silica gel column chromatography to obtain 290 mg of an off-white solid. 1H NMR (400MHz, DMSO-d6): δ10.40 (s, 1H), 9.08 (d, J = 7.6Hz, 1H), 8.95 (s, 1H), 8.0 2(d,J=4.8Hz,1H),7.60-7.57(m,2H),7.35(d,J=8.4Hz,1H),7.01(d,J=4.8Hz, 1H),6.93(t,J=4.4Hz,1H),6.70-6.66(m,1H),4.78-4.71(m,1H),4.08(s,2H), 3.23(d,J=2.0Hz,1H),2.53-2.51(m,3H),1.34(d,J=7.2Hz,3H).ESI-MS:[M+H] + =662.0.

[0217] Example 5

[0218] To a 100 mL single-necked flask, add N-ethyl-2-oxoxazoline-3-sulfonamide (1.4 g), 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-N-(alkyn-2-prop-1-yl)benzamide (400 mg), triethylamine (800 mg), and 1,2-dichloroethane (40 mL). The reaction system was heated to 80°C for 4.5 hours. After cooling to room temperature, the mixture was poured into a water / tetrahydrofuran mixture (90 / 10, v / v) and extracted with ethyl acetate. The organic phases were combined, dried, and concentrated. The crude product was purified by silica gel column chromatography to obtain the crude product. The crude product was washed with a n-hexane / acetone mixture (4 / 1, v / v) to obtain 150 mg of an off-white solid. 1 H NMR (400MHz, DMSO-d6): δ10.40(s,1H),9.17(t,J=5.2Hz,1H),9.06(s,1H),8.02(d,J=4 .8Hz,1H),7.59(dd,J1=10.8Hz,J2=1.6Hz,1H),7.55(d,J=7.2Hz,1H),7.37(d,J=8.4Hz, 1H),7.18-7.15(m,1H),6.93(t,J=4.4Hz,1H),6.75-6.68(m,1H),4.07(s,2H),4.02-4.0 0(m,2H),3.17(t,J=2.8Hz,1H),2.96-2.89(m,2H),1.02(t,J=7.2Hz,3H).ESI-MS:[M+H] + =662.1.

[0219] Example 6

[0220] To a 100 mL single-necked flask, add N-cyclopropyl-2-oxoxazoline-3-sulfonamide (1.5 g), 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-N-(prop-2-yn-1-yl)benzamide (400 mg), triethylamine (729 mg), and 1,2-dichloroethane (20 mL). The reaction system was heated to 80°C for 6 h. After cooling to room temperature, the mixture was poured into a water / tetrahydrofuran mixture (90 / 10, v / v). The aqueous phase was extracted with ethyl acetate, and the organic phases were combined, dried, concentrated, and purified by silica gel column chromatography to obtain the crude product. The crude product was washed with a n-hexane / acetone mixture (4 / 1, v / v) to obtain 100 mg of a yellow solid. 1 H NMR (400MHz, DMSO-d6): δ10.55(s,1H),9.18(t,J=5.6Hz,1H),9.06(s,1H), 8.03(d,J=4.8Hz,1H),7.62-7.55(m,3H),7.38-7.36(m,1H),6.95(t,J=4.8H z,1H),6.74-6.68(m,1H),4.08(s,2H),4.02-4.00(m,2H),3.17(t,J=6.4Hz, 1H),2.34-2.28(m,1H),0.64-0.58(m,2H),0.57-0.50(m,2H).ESI-MS:[M+H] + =674.0.

[0221] Example 7

[0222] Step 1:

[0223] In a 100 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoic acid hydrochloride (600 mg), 1-hydroxybenzotriazole (297 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (844 mg) and N,N-dimethylformamide (32 mL) were added, and the system was stirred at room temperature for 2 hours. Subsequently, pent-4-yn-1-amine hydrochloride (263 mg) and N,N-diisopropylethylamine (711 mg) were added to the system, and the reaction was continued to stir at room temperature for 2 hours. The reaction system was added to a saturated sodium bicarbonate aqueous solution to precipitate a solid. The filter cake was filtered and washed with ethyl acetate / petroleum ether (1 / 3) to obtain 500 mg of a light yellow solid. ESI-MS: [M+H] + =583.1.

[0224] Step 2:

[0225] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-N-(cyanomethyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzamide (500 mg), N,N-dimethylformamide (10 mL) and pyridine (595 mg) were added, and the system was placed in an ice bath and stirred. When the system dropped below 0°C, methylaminosulfonyl chloride (700 mg) was slowly added dropwise. The reaction was stopped after 30 minutes, and the system was poured into a saturated aqueous sodium bicarbonate solution and stirred at room temperature for 10 minutes. The mixture was extracted with ethyl acetate, and the organic phases were combined and dried over anhydrous sodium sulfate. The solvent was removed by filtration and then the crude product was purified by silica gel column chromatography to obtain 330 mg of an off-white solid. 1 H NMR (400MHz, DMSO-d6): δ10.40(s,1H),9.05(s,1H),8.70(t,J=1.2Hz,1H),8.02(d,J=4.8Hz,1H),7 .58(dd,J1=10.8Hz,J2=2.0Hz,1H),7.55(d,J=8.0Hz,1H),7.36(dd,J1=8.4Hz,J2=1.2Hz,1H),7.03 -7.00(m,1H),6.93(t,J=4.8Hz,1H),6.71-6.65(m,1H),4.08(s,2H),3.29-3.24(m,2H),2.78(t,J= 2.8Hz,1H),2.53-2.52(m,3H),2.19(td,J1=7.2Hz,J2=2.8Hz,2H),1.68-1.61(m,2H).ESI-MS:[M+H] + =676.1.

[0226] Example 8

[0227] Step 1:

[0228] To a 100 mL single-necked flask, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoic acid hydrochloride (600 mg), 1-hydroxybenzotriazole (297 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (844 mg), and N,N-dimethylformamide (32 mL) were added and the reaction was stirred at room temperature for 2 hours. Subsequently, but-2-yn-1-amine hydrochloride (232 mg) and N,N-diisopropylethylamine (711 mg) were added to the system, and the reaction was stirred at room temperature for another 2 hours. The reaction system was added to saturated aqueous sodium bicarbonate solution to precipitate a solid. The filter cake was filtered and washed with ethyl acetate / petroleum ether (1 / 3) to obtain 524 mg of a light yellow solid. 1 H NMR (400MHz, DMSO-d6): δ9.08(t,J=1.2Hz,1H),9.05(s,1H),7.66(d,J=5.2Hz,1H),7.59(dd,J1=10.8Hz,J2=2.0Hz,1H),7.53(d,J=6.8Hz,1H),7 .36(dd,J1=8.4Hz,J2=1.2Hz,1H),6.70-6.65(m,1H),6.40(t,J=4.8Hz,1H),6.17(s,2H),3.98-3.96(m,4H),1.77(t,J=2.4Hz,3H).ESI-MS:[M+H] + =569.0.

[0229] Step 2:

[0230] In a 50 mL single-necked flask, (R)-5-((2-amino-3-fluoropyridin-4-yl)methyl)-N-(but-2-yn-1-yl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzamide (500 mg), N,N-dimethylformamide (10 mL) and pyridine (595 mg) were added, and the system was placed in an ice bath and stirred. When the system temperature dropped below 0°C, methylaminosulfonyl chloride (700 mg) was slowly added dropwise. The reaction was stopped after 30 minutes, and the system was poured into a saturated aqueous sodium bicarbonate solution and stirred at room temperature for 10 minutes. Extracted with ethyl acetate, the organic phases were combined and dried over anhydrous sodium sulfate. Filtered and the solvent was removed by rotation, and the crude product was purified by silica gel column chromatography to obtain 340 mg of an off-white solid. 1H NMR (400MHz, DMSO-d6): δ10.39(s,1H),9.08(s,2H),8.02(d,J=5.2Hz,1H),7.61-7.55(m,2H),7.37(dd,J1=8.4Hz,J2=1.2Hz,1H),7.01(q,J= 4.8Hz,1H),6.95(t,J=4.8Hz,1H),6.73-6.67(m,1H),4.07(s,2H),3.99-3.97(m,2H),2.53-2.51(m,3H),1.77(t,J=2.4Hz,3H).ESI-MS:[M+H] + =662.0.

[0231] Example 9

[0232] Step 1:

[0233] In a 100 mL single-necked flask, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoic acid hydrochloride (1.3 g), 1-hydroxybenzotriazole (867 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.5 g), and N,N-dimethylformamide (20 mL) were added and the reaction was stirred at room temperature for 2 h. Propargylamine (354 mg) and N,N-diisopropylethylamine (2.1 g) were then added to the system, and the reaction was stirred at room temperature for another 2 h. The reaction system was added to a saturated aqueous sodium bicarbonate solution to precipitate a solid. The filter cake was filtered and washed with ethyl acetate / petroleum ether (1 / 3) to obtain 900 mg of a light yellow solid. ESI-MS: [M+H] + =443.2.

[0234] Step 2:

[0235] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-methylphenyl)amino)-N-(prop-2-yn-1-yl)benzamide (500 mg), N,N-dimethylformamide (6 mL) and pyridine (894 mg) were added, and the system was placed in an ice bath and stirred. When the system dropped below 0°C, methylaminosulfonyl chloride (732 mg) was slowly added dropwise. After 30 minutes, the reaction was stopped, and the system was poured into a saturated aqueous sodium bicarbonate solution and stirred at room temperature for 10 minutes. Extracted with ethyl acetate, the organic phases were combined and dried over anhydrous sodium sulfate. Filtered and the solvent was removed by rotation, and the crude product was purified by silica gel column chromatography to obtain 300 mg of an off-white solid. 1H NMR (400MHz, DMSO-d6): δ10.39(s,1H),9.17(t,J=5.2Hz,1H),9.08(s,1H),8.02(d,J=4.8Hz,1H),7.56(d,J=7.2Hz,1H),7.05 -7.02(m,2H),6.93(s,1H),6.88-6.81(m,2H),4.05-4.02(m,4H),3.16(s,1H),2.53-2.51(m,3H),2.25(s,3H).ESI-MS:[M+H] + =536.2.

[0236] Example 10

[0237] Step 1:

[0238] In a 100 mL single-necked bottle, methyl 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoate (1.5 g), cuprous iodide (11 mg), triethylamine (4.5 mL), tetrahydrofuran (15 mL) and bistriphenylphosphine palladium dichloride (40 mg) were added, and the system was replaced with a nitrogen environment and stirred at room temperature for reaction. Trimethylsilyl acetylene (1.1 g) was then slowly added dropwise to the system. After the addition, the reaction system was stirred at room temperature for 21 hours. The reaction system was diluted with ethyl acetate, filtered, the filtrate was collected, washed with saturated brine, the organic phase was collected, and dried over anhydrous sodium sulfate. Filter and spin-dry the solvent, and the crude product was purified by silica gel column chromatography to obtain 1.0 g of an off-white solid. ESI-MS: [M+H] + =502.2.

[0239] Step 2:

[0240] In a 50 mL single-necked bottle, methyl 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-((trimethylsilyl)ethynyl)phenyl)amino)benzoate (1.0 g), tetrahydrofuran (15 mL) and water (7.5 mL) were added, and the system was stirred at room temperature until clear. The system was then placed in an ice bath, and lithium hydroxide monohydrate (252 mg) was slowly added to the system. After the addition, the system was warmed to room temperature and stirred for 16.5 hours. Hydrochloric acid (6 M, 6 mL) was then slowly added dropwise to the system to adjust the pH of the system to 1-2 and stirred for 5 minutes. The tetrahydrofuran in the system was removed under reduced pressure, and then water was added to dilute the concentrate and stirred for 10 minutes. The filter cake was collected by filtration and washed with water and tert-methyl ether respectively to obtain 810 mg of a brown solid. ESI-MS: [M+H] + =416.1.

[0241] Step 3:

[0242] In a 50 mL single-necked flask, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-2-((4-ethynyl-2-fluorophenyl)amino)-3,4-difluorobenzoic acid hydrochloride (870 mg), 1-hydroxybenzotriazole (513 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5 g), and N,N-dimethylformamide (15 mL) were added and the reaction was stirred at room temperature for 2 hours. Propargylamine (209 mg) and N,N-diisopropylethylamine (1.3 g) were then added to the system, and the reaction was stirred at room temperature for another 2 hours. The reaction system was added to a saturated aqueous sodium bicarbonate solution to precipitate a solid. The filter cake was filtered and purified by silica gel column chromatography to yield 750 mg of a light brown solid. 1 H NMR (400MHz, DMSO-d6): δ9.16(t,J=5.2Hz,1H),9.08(s,1H),7.67(d,J=5.2Hz,1H),7.53( d,J=6.8Hz,1H),7.34(d,J=12.0Hz,1H),7.16(d,J=8.4Hz,1H),6.84-6.78(m,1H),6.41(t J=4.8Hz,1H),6.17(s,2H),4.12(s,1H),4.023.97(s,4H),3.15(s,1H).ESI-MS:[M+H] + =453.1.

[0243] Step 4:

[0244] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-2-((4-ethynyl-2-fluoropyridinyl)amino)-3,4-difluoro-N-(prop-2-yn-1-yl)benzamide (720 mg), N,N-dimethylformamide (9 mL) and pyridine (1.3 g) were added, and the system was placed in an ice bath and stirred. When the system dropped below 0 ° C, methylaminosulfonyl chloride (900 mg) was slowly added dropwise. After 30 minutes, the reaction was stopped, and the system was poured into a saturated sodium bicarbonate aqueous solution and stirred at room temperature for 10 minutes. Extracted with ethyl acetate, the organic phases were combined and dried over anhydrous sodium sulfate. Filtered and the solvent was removed by rotation, and the crude product was then purified by silica gel column chromatography to obtain 700 mg of an off-white solid. 1H NMR (400MHz, DMSO-d6): δ10.40(s,1H),9.16(t,J=5.6Hz,1H),9.12(s,1H),8.03 (d,J=5.2Hz,1H),7.56(d,J=7.2Hz,1H),7.34(d,J=12.8Hz,1H),7.16(d,J=8.4H z,1H),7.04-7.00(m,1H),6.95(t,J=4.4Hz,1H),6.87-6.81(m,1H),4.13(s,1H) ,4.09(s,2H),4.02-4.01(m,2H),3.16(s,1H),2.53-2.52(m,3H).ESI-MS:[M+H] + =546.1.

[0245] Example 11

[0246] Step 1:

[0247] In a 100 mL single-necked flask, methyl 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoate (1.9 g), potassium cyclopropylfluoroborate (799 mg), cesium carbonate (3.5 g), toluene (38 mL), water (4 mL), and [1,1′-bis(diphenylphosphino)ferrocene]palladium dichloride (294 mg) were added and the atmosphere was replaced with nitrogen. The temperature was raised to 100°C and stirred for 23 hours. The reaction system was cooled to room temperature, ethyl acetate was added, and the reaction system was washed with saturated brine. The organic phase was collected and dried over anhydrous sodium sulfate. The solvent was removed by filtration, and the concentrate was purified by silica gel column chromatography to obtain 1.0 g of a yellow solid. ESI-MS: [M+H] + =446.2.

[0248] Step 2:

[0249] In a 50 mL single-necked bottle, methyl 5-((2-amino-3-fluoropyridin-4-yl)methyl)-2-((4-cyclopropyl-2-fluorophenyl)amino)-3,4-difluorobenzoate (1.0 g), tetrahydrofuran (14 mL) and water (7 mL) were added, and the system was stirred at room temperature until clear. The system was then placed in an ice bath, and lithium hydroxide monohydrate (283 mg) was slowly added to the system. After the addition, the system was warmed to room temperature and stirred for 19 hours. Hydrochloric acid (6 M, 6 mL) was slowly added dropwise to the system to adjust the pH to 1-2 and stirred for 5 minutes. The tetrahydrofuran in the system was removed by concentration under reduced pressure, and then the concentrate was diluted with water and stirred for 10 minutes. Filter, collect the filter cake, and wash with water and tert-methyl ether respectively to obtain 1.0 g of an off-white solid. ESI-MS: [M+H]+ =432.2.

[0250] Step 3:

[0251] In a 50 mL single-necked flask, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-2-((4-cyclopropyl-2-fluorophenyl)amino)-3,4-difluorobenzoic acid hydrochloride (1.0 g), 1-hydroxybenzotriazole (639 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (805 mg), and N,N-dimethylformamide (10 mL) were added and the reaction was stirred at room temperature for 2 hours. Propargylamine (347 mg) and N,N-diisopropylethylamine (814 mg) were then added to the system, and the reaction was stirred at room temperature for another 2 hours. The reaction system was added to a saturated aqueous sodium bicarbonate solution to precipitate a solid. The filter cake was filtered, and purified by silica gel column chromatography to yield 0.9 g of an off-white solid. 1 H NMR (400MHz, DMSO-d6): δ9.16(t,J=5.2Hz,1H),9.05(s,1H),7.66(d,J=4.8Hz,1H),7.53(d,J=7.2Hz,1H),6.90(d,J=12.8Hz,1H),6.80(s,2H),6.3 9(t,J=4.8Hz,1H),6.16(s,2H),4.02-4.01(m,2H),3.93(s,2H),3.15(s,1 H),1.91-1.84(m,1H),093-0.88(m,2H),0.64-0.61(m,2H).ESI-MS:[M+H] + =469.2.

[0252] Step 4:

[0253] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-2-((4-cyclopropyl-2-fluorophenyl)amino)-3,4-difluoro-N-(prop-2-yn-1-yl)benzamide (1.0 g), N,N-dimethylformamide (10 mL) and pyridine (1.7 g) were added, and the system was placed in an ice bath and stirred. When the system dropped below 0°C, methylaminosulfonyl chloride (830 mg) was slowly added dropwise. After 30 minutes, the reaction was stopped, and the system was poured into a saturated aqueous sodium bicarbonate solution and stirred at room temperature for 10 minutes. The mixture was extracted with ethyl acetate, the organic phases were combined, and dried over anhydrous sodium sulfate. The solvent was filtered and spun off, and the crude product was then purified by silica gel column chromatography to obtain 400 mg of an off-white solid. 1H NMR (400MHz, DMSO-d6): δ10.39(s,1H),9.16(t,J=5.2Hz,1H),9.08(s,1H),8.0 2(d,J=4.0Hz,1H),7.55(d,J=7.2Hz,1H),7.01(q,J=4.8Hz,1H),6.94-6.89(m,2 H),6.83-6.79(m,2H),4.05(s,2H),4.02-4.01(m,2H),3.16(s,1H),2.53-2.51 (m,3H),1.91-1.84(m,1H),0.93-0.89(m,2H),0.65-0.61(m,2H).ESI-MS:[M+H] + =562.2.

[0254] Example 12

[0255] Step 1:

[0256] In a 50 mL single-necked bottle, methyl 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoate (1.3 g), potassium vinyl fluoroborate (636 mg), potassium carbonate (984 mg), 1,4-dioxane (12 mL), water (4 mL) and [1,1′-bis(diphenylphosphino)ferrocene]palladium dichloride (86 mg) were added. The atmosphere was replaced with nitrogen and the temperature was raised to 95°C and stirred for 2 h. After the system was cooled to room temperature, it was poured into water to quench the reaction. The organic phases were extracted with ethyl acetate, combined, and dried over anhydrous sodium sulfate. The solvent was filtered and removed by rotation to obtain 950 mg of a brown solid. ESI-MS: [M+H] + =432.2.

[0257] Step 2:

[0258] In a 50 mL single-necked flask, methyl 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-vinylphenyl)amino)benzoate (950 mg), tetrahydrofuran (20 mL), and water (10 mL) were added and stirred at room temperature until the solution became clear. The system was then placed in an ice bath and lithium hydroxide monohydrate (360 mg) was slowly added. After the addition, the system was warmed to room temperature and stirred for 3 hours. Hydrochloric acid (6 M, 5 mL) was then slowly added dropwise to the system to adjust the pH to 1-2 and stirred for 5 minutes. The system was concentrated under reduced pressure to remove tetrahydrofuran, and the concentrate was diluted with water and stirred for 10 minutes. The filter cake was collected by filtration and washed with water and tertiary methyl ether to obtain 700 mg of a brown solid. The product from this step was used directly in the next step without further characterization or purification.

[0259] Step 3:

[0260] In a 50 mL single-necked flask, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-vinylphenyl)amino)benzoic acid hydrochloride (700 mg), 1-hydroxybenzotriazole (396 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.1 g) and N,N-dimethylformamide (15 mL) were added, and the system was stirred at room temperature for 2 hours. Subsequently, propargylamine (162 mg) and N,N-diisopropylethylamine (598 mg) were added to the system, and the reaction was continued to stir at room temperature for 2 hours. The reaction system was added to a saturated sodium bicarbonate aqueous solution to precipitate a solid. The filter cake was filtered and washed with ethyl acetate / petroleum ether (1 / 3) to obtain 460 mg of a light yellow solid. ESI-MS: [M+H] + =455.2.

[0261] Step 4:

[0262] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-vinylphenyl)amino)-N-(prop-2-yn-1-yl)benzamide (460 mg), N,N-dimethylformamide (10 mL) and pyridine (801 mg) were added, and the system was placed in an ice bath and stirred. When the system dropped below 0°C, methylaminosulfonyl chloride (600 mg) was slowly added dropwise. After 30 minutes, the reaction was stopped, and the system was poured into a saturated aqueous sodium bicarbonate solution (50 mL) and stirred at room temperature for 10 minutes. Extracted with ethyl acetate, the organic phases were combined and dried over anhydrous sodium sulfate. Filtered and the solvent was removed by rotation, and the crude product was then purified by silica gel column chromatography to obtain 300 mg of an off-white solid. 1H NMR (400MHz, DMSO-d6): δ10.40(s,1H),9.18(t,J=5.2Hz,1H),9.14(s,1H),8.01(d,J=4.4Hz, 1H),7.57(d,J=7.2Hz,1H),7.39(d,J=12.8Hz,1H),7.14(d,J=8.1Hz,1H),7.02(s,1H)6.94(s, 1H),6.90-6.84(m,1H),6.65(dd,J1=17.6Hz,J2=11.2Hz,1H),5.75(d,J=17.6Hz,1H),5.18(d, J=10.8Hz,1H),4.07(s,2H),4.04-4.01(m,2H),3.16(s,1H),2.52-2.51(m,3H).ESI-MS:[M+H] + =548.1.

[0263] Example 13

[0264] Step 1:

[0265] In a 100 mL single-necked flask, methyl 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-vinylphenyl)amino)benzoate (1.1 g), potassium osmate dihydrate (91 mg), sodium periodate (2.7 g), 1,4-dioxane (30 mL), and water (12 mL) were added and stirred at room temperature for 2 h. The reaction mixture was poured into water and extracted with ethyl acetate. The organic phases were combined and dried over anhydrous sodium sulfate. The mixture was filtered and the solvent removed by vortexing to yield 950 mg of a solid. The solid was diluted with acetic acid (20 mL), followed by the addition of iron powder (9.0 g) and the reaction was stirred at room temperature for 1 h. Ethyl acetate was added, and the insoluble material was removed by filtration. The filtrate was collected and concentrated, and the concentrate was diluted with water. The pH of the mixture was adjusted to 7-8 with aqueous sodium bicarbonate solution. The mixture was extracted with ethyl acetate, and the organic phases were combined and dried over anhydrous sodium sulfate. The crude product was filtered and the solvent was removed by rotary evaporation, and then purified by silica gel column chromatography to obtain 450 mg of a yellow solid. ESI-MS: [M+H] + =434.1.

[0266] Step 2:

[0267] In a 50 mL single-necked flask, methyl 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-formylphenyl)amino)benzoate (450 mg), tetrahydrofuran (10 mL) and water (5 mL) were added and the system was stirred at room temperature until clear. The system was then placed in an ice bath and lithium hydroxide monohydrate (131 mg) was slowly added to the system. After the addition was complete, the system was warmed to room temperature and stirred for 2 hours. Hydrochloric acid (6 M, 3 mL) was then slowly added dropwise to the system to adjust the pH to 1-2 and stirred for 5 minutes. The tetrahydrofuran in the system was removed under reduced pressure, and the concentrate was then diluted with water and stirred for 10 minutes. The filter cake was collected by filtration and washed with water and tertiary methyl ether respectively to obtain 390 mg of a light yellow solid. This step was not further purified or characterized and the next step was directly carried out.

[0268] Step 3:

[0269] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-formylphenyl)amino)benzoic acid hydrochloride (390 mg), 1-hydroxybenzotriazole (220 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (607 mg) and N,N-dimethylformamide (12 mL) were added, and the system was stirred at room temperature for 2 hours. Subsequently, propargylamine (90 mg) and N,N-diisopropylethylamine (332 mg) were added to the system, and the reaction was continued to stir at room temperature for 1 hour. The reaction system was added to a saturated sodium bicarbonate aqueous solution, and the precipitated solid was filtered to obtain 250 mg of a yellow solid. ESI-MS: [M+H] + =457.1.

[0270] Step 4:

[0271] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-2-((2-fluoro-4-formylphenyl)amino)-3,4-difluoro-N-(prop-2-yn-1-yl)benzamide (120 mg), N,N-dimethylformamide (2 mL) and pyridine (237 mg) were added, and the system was placed in an ice bath and stirred. When the system dropped below 0°C, methylaminosulfonyl chloride (700 mg) was slowly added dropwise. After 30 minutes, the reaction was stopped, and the system was poured into a saturated aqueous sodium bicarbonate solution and stirred at room temperature for 10 minutes. The mixture was extracted with ethyl acetate, and the organic phases were combined and dried over anhydrous sodium sulfate. The solvent was removed by filtration, and the crude product was then purified by silica gel column chromatography to obtain 70 mg of an off-white solid. 1H NMR (400MHz, DMSO-d6): δ10.41(s,1H),9.79(s,1H),9.19(s,1H),9.12(t,J=5.2Hz,1H),8.04(d,J=5.2Hz,1H),7.67(d,J=11.6Hz,1H),7.61(d, J=8.4Hz,1H),7.55(d,J=6.8Hz,1H),7.04-6.90(m,3H),4.13(s,2H),4.00-3.99(m,2H),3.14(s,1H),2.53-2.52(d,J=5.2Hz,3H).ESI-MS:[M+H] + =550.1.

[0272] Example 14

[0273] Step 1:

[0274] In a 100 mL single-necked flask, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-(2-fluoro-4-methoxyphenyl)benzoic acid hydrochloride (300 mg), propargylamine (40 mg), N,N-diisopropylethylamine (272 mg), and N,N-dimethylformamide (6 mL) were added. The system was then replaced with a nitrogen atmosphere, and O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate (228 mg) was added portionwise. The mixture was stirred at room temperature for 3 hours. The reaction solution was added to semi-saturated brine and extracted with ethyl acetate. The organic phases were combined, washed with aqueous citric acid and saturated brine, dried, and concentrated. The crude product was purified by silica gel column chromatography to obtain 167 mg of a yellow solid. ESI-MS: [M+H] + =459.1.

[0275] Step 2:

[0276] In a 50 mL single-necked bottle, add N-propargyl-5-((2-amino-3-fluoropyridin-4-yl)methyl)-2-((4-methoxy-2-fluorophenyl)amino)-3,4-difluorobenzamide (150 mg), N,N-dimethylformamide (6 mL) and pyridine (209 mg), and place the system in an ice bath and stir. When the system drops below 0°C, slowly add methylaminosulfonyl chloride (257 mg) dropwise and react for 2 hours. Add water to the system and stir at room temperature for 10 minutes. Extract with ethyl acetate, combine the organic phases, and dry over anhydrous sodium sulfate. Filter and spin off the solvent, and the crude product is purified by silica gel column chromatography to obtain 62 mg of a light yellow solid. 1H NMR (400MHz, DMSO-d6): δ10.41 (s, 1H), 9.16 (t, J = 5.2Hz, 1H), 9.11 (s, 1H),8.02(d,J=5.2Hz,1H),7.55(d,J=7.2Hz,1H),7.04-6.86(m,4H),6.69-6.66(m,1H), 4.02-4.01(m,4H),3.73(s,3H),3.17(t,J=2.4Hz,1H),2.52-2.51(m,3H).ESI-MS:[M+H] + =552.1.

[0277] Example 15

[0278] Step 1:

[0279] In a 100 mL single-necked flask, 5-(3-amino-2-fluorobenzyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoic acid hydrochloride (310 mg), 1-hydroxybenzotriazole (170 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (322 mg), and N,N-dimethylformamide (6 mL) were added, and the system was stirred at room temperature for 2 hours. Subsequently, propargylamine (93 mg) and N,N-diisopropylethylamine (217 mg) were added to the system, and the reaction was continued to stir at room temperature for 2 hours. The reaction system was added to a saturated sodium bicarbonate aqueous solution, and the precipitated solid was filtered with suction. The filter cake was purified by silica gel column chromatography to obtain 220 mg of an off-white solid. ESI-MS: [M+H] + =554.0.

[0280] Step 2:

[0281] In a 50 mL single-necked bottle, 5-(3-amino-2-fluorobenzyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-N-(alkyn-2-prop-1-yl)benzamide (220 mg), N,N-dimethylformamide (5 mL) and pyridine (312 mg) were added and the system was placed in an ice bath and stirred. After the system cooled to below 0°C, methylaminosulfonyl chloride (156 mg) was slowly added dropwise. After 30 minutes, the reaction was stopped and the system was poured into a saturated aqueous sodium bicarbonate solution and stirred at room temperature for 10 minutes. The organic phases were extracted with ethyl acetate, combined, and dried over anhydrous sodium sulfate. The solvent was filtered and removed by rotation. The crude product was purified by silica gel column chromatography to obtain 150 mg of a light yellow solid. 1H NMR (400MHz, DMSO-d6): δ9.43 (s, 1H), 9.15 (t, J = 5.6Hz, 1H), 9.00 (s, 1H), 7.59 (dd ,J1=10.8Hz,J2=1.6Hz,1H),7.52(d,J=6.8Hz,1H),7.38-7.31(m,2H),7.25(q,J=4. 8Hz,1H),7.11(t,J=8.0Hz,1H),7.02(t,J=6.8Hz,1H),6.70-6.65(m,1H),4.03(s,2 H),4.01-3.99(m,2H),3.16(t,J=2.4Hz,1H),2.52(d,J=5.2Hz,3H).ESI-MS:[M+Na] + =669.0.

[0282] Example 16

[0283] Step 1:

[0284] In a 100 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-(phenylamino)benzoic acid hydrochloride (350 mg), 1-hydroxybenzotriazole (230 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (656 mg) and N,N-dimethylformamide (5 mL) were added, and the system was stirred at room temperature for 2 hours. Subsequently, propargylamine (100 mg) and N,N-diisopropylethylamine (548 mg) were added to the system, and the reaction was continued to stir at room temperature for 2 hours. The reaction system was added to a saturated sodium bicarbonate aqueous solution to precipitate a solid. The filter cake was filtered and purified by silica gel column chromatography to obtain 200 mg of a light yellow solid. ESI-MS: [M+H] + =411.2.

[0285] Step 2:

[0286] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-(phenylamino)-N-(prop-2-yn-1-yl)benzamide (200 mg), N,N-dimethylformamide (10 mL) and pyridine (386 mg) were added, and the system was placed in an ice bath and stirred. When the system dropped below 0°C, methylaminosulfonyl chloride (190 mg) was slowly added dropwise. After 30 minutes, the reaction was stopped, and the system was poured into a saturated aqueous sodium bicarbonate solution and stirred at room temperature for 10 minutes. Extracted with ethyl acetate, the organic phases were combined and dried over anhydrous sodium sulfate. Filtered and the solvent was removed by rotation, and the crude product was purified by silica gel column chromatography to obtain 160 mg of an off-white solid. 1H NMR (400MHz, DMSO-d6): δ10.41(s,1H),9.08(t,J=5.6Hz,1H),8.86(s,1H),8.03(d,J=5.2Hz,1H),7.51(d,J=6.4Hz,1H),7.23-7.19(m,2H),7.03(q ,J=5.2Hz,1H),6.95(t,J=5.2Hz,1H),6.90-6.84(m,3H),4.01(s,2H),3. 99-3.97(m,2H),3.15(t,J=2.4Hz,1H),2.52-2.51(m,3H).ESI-MS:[M+Na] + =526.1.

[0287] Example 17

[0288] Step 1:

[0289] To a 3L reaction flask, add 1000mL of dichloromethane, trimethylsilyl trifluoromethanesulfonate (271.8g), triethylamine (123.7g), and silver trifluoromethanesulfonate (10g). Slowly add a solution of ethyl propargyl acid (100g) in dichloromethane (100mL) dropwise. After the addition is complete, stir at room temperature for 30 minutes and concentrate under reduced pressure to remove the dichloromethane. Extract the concentrated residue with n-hexane. Combine the n-hexanes and wash with saturated ammonium chloride solution. Dry the n-hexane phase over anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain approximately 140g of a light yellow liquid.

[0290] Step 2:

[0291] In a 1000mL three-necked flask, add ethyl 3-(trimethylsilyl)propionate (50g), 500mL of ether, sodium borodeuteride (17.3g), lithium chloride (17.4g), and heavy water (8.2g). Stir at room temperature for 40 minutes, then warm to 33°C and react for 8 hours. Slowly add the reaction solution to 500mL of 10% aqueous acetic acid. Extract with n-hexane, separate the layers, and extract the aqueous layer again with n-hexane. Combine the organic phases, wash with 1N hydrochloric acid solution, and then wash the organic phase with water until neutral. Dry over anhydrous sodium sulfate and concentrate under reduced pressure to obtain a light yellow liquid. 1 H NMR (400MHz, DMSO-d6): δ5.20 (brs, 1H), 0.13 (s, 9H).

[0292] Step 3:

[0293] In a 100 mL single-necked flask, add 3-(trimethylsilyl)prop-2-yn-1,1-d2-1-ol (6.0 g) and dichloromethane (75 mL), and cool the system to -10°C. Phosphorus tribromide (30.0 mg) is then added dropwise. After addition, the reaction system is warmed to room temperature for 1 hour. The system is poured onto ice, and the pH is adjusted to neutral with saturated sodium bicarbonate aqueous solution. Extract with dichloromethane, combine the organic phases, dry, and concentrate to yield 5.1 g of a colorless, transparent liquid.

[0294] Step 4:

[0295] In a 250 mL single-necked flask, add (3-bromoprop-1-yn-1-yl-3,3-d2)trimethylsilane (10.0 g), potassium phthalimide (10.5 g), potassium carbonate (8.6 g), and N,N-dimethylformamide (100 mL). Stir the reaction at room temperature for 16 hours. Pour the mixture into water, extract with ethyl acetate, combine the organic phases, and dry over anhydrous sodium sulfate. Filter and concentrate to obtain 12.0 g of an off-white solid.

[0296] Step 5:

[0297] In a 250 mL single-necked bottle, 2-((3-(trimethylsilyl)prop-2-yn-1-yl-1,1-d2)oxy)isoindoline-1,3-dione (5.0 g) and dichloromethane / methanol (2 / 1, v / v, 96 mL in total) were added, and the system was replaced with a nitrogen atmosphere. Hydrazine hydrate (1.4 g) was slowly added dropwise to the system. After the addition, the reaction was stirred for 20 h. Filtered with suction, the filter cake was washed with dichloromethane, and the organic phases were combined. Concentrated, the concentrate was diluted with tert-methyl ether, filtered, and the pH of the filtrate was adjusted to 1 with a solution of hydrogen chloride in ethyl acetate. The filtrate was concentrated to obtain 1.8 g of a light yellow oil.

[0298] Step 6:

[0299] In a 100 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoic acid hydrochloride (770 mg), O-benzotriazole-N,N,N',N',-tetramethyluronium tetrafluoroborate (311 mg), N,N-diisopropylethylamine (188 mg), O-(3-(trimethylsilyl)prop-2-yn-1-yl-1,1-d2)hydroxylamine hydrochloride (165 mg) and N,N-dimethylformamide (10 mL) were added, and the system was stirred at room temperature for 3 hours. The reaction system was added to saturated brine, extracted with ethyl acetate, and the organic phases were combined and dried over anhydrous sodium sulfate. Filtered and concentrated to give 550 mg of a brown solid. ESI-MS: [M+H] + =629.1.

[0300] Step 7:

[0301] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-N-(3-(trimethylsilyl)prop-2-yn-1-yl-1,1-d2)benzamide (795 mg), tetrahydrofuran (30 mL) and tetrabutylammonium fluoride trihydrate (998 mg) were added and stirred at room temperature for 10 minutes. The system was poured into water and extracted with ethyl acetate. The organic phases were combined and dried over anhydrous sodium sulfate. The product was concentrated to give 300 mg of a white solid. ESI-MS: [M+H] + =557.0.

[0302] Step 8:

[0303] In a 50 mL single-necked flask, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-N-(prop-2-yn-1-yl-1,1-d2)benzamide (336 mg), N,N-dimethylformamide (8 mL), and pyridine (763 mg) were added and the system was stirred in an ice bath. After the system cooled to below 0°C, methylaminosulfonyl chloride (777 mg) was slowly added dropwise. The reaction was stopped after 30 minutes, and the system was poured into a saturated aqueous sodium bicarbonate solution and stirred at room temperature for 10 minutes. The mixture was extracted with ethyl acetate, the organic phases were combined, and dried over anhydrous sodium sulfate. The solvent was removed by filtration, and the crude product was purified by silica gel column chromatography to obtain 180 mg of a light yellow solid. 1 H NMR (400MHz, DMSO-d6): δ10.42(s,1H),9.16(s,1H),9.07(s,1H),8.02(d,J=4.8Hz,1H),7.61-7.55(m,2H),7.37(d,J=8.4Hz,1 H),7.33(q,J=4.8Hz,1H),6.95(t,J=4.8Hz,1H),6.74-6.68(m,1H),4.07(s,2H),3.16(s,1H),2.52-2.51(m,3H).ESI-MS:[M+H] + =650.0.

[0304] Example 18

[0305] Step 1:

[0306] In a 100 mL single-necked flask, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoic acid hydrochloride (600 mg), 1-hydroxybenzotriazole (297 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (844 mg), and N,N-dimethylformamide (32 mL) were added, and the system was stirred at room temperature for 2 hours. Subsequently, O-(prop-2-yn-1-yl)hydroxylamine (237 mg) and N,N-diisopropylethylamine (711 mg) were added to the system, and the reaction was continued to stir at room temperature for 2 hours. The reaction system was added to saturated sodium bicarbonate aqueous solution to precipitate a solid. The filter cake was filtered and washed with ethyl acetate / petroleum ether (1 / 3) to obtain 500 mg of a light yellow solid. ESI-MS: [M+H] + =571.0.

[0307] Step 2:

[0308] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-N-(prop-2-yn-1-yloxy)benzamide (500 mg), N,N-dimethylformamide (10 mL) and pyridine (696 mg) were added, and the system was placed in an ice bath and stirred. When the system dropped below 0°C, methylaminosulfonyl chloride (700 mg) was slowly added dropwise. After the addition was completed and the reaction was stopped after 30 minutes, the system was poured into a saturated aqueous sodium bicarbonate solution and stirred at room temperature for 10 minutes. Extracted with ethyl acetate, the organic phases were combined and dried over anhydrous sodium sulfate. Filtered and the solvent was removed by rotation, and the crude product was purified by silica gel column chromatography to obtain 381 mg of an off-white solid. 1 H NMR (400MHz, DMSO-d6): δ12.02(s,1H),10.42(s,1H),8.53(s,1H),8.02(d,J=5.2Hz,1H),7.57(dd,J1=10.8Hz,J2=2.4Hz,1H),7.36-7.35(m, 2H),7.04(q,J=4.8Hz,1H),6.95(t,J=4.8Hz,1H),6.70-6.65(m,1H),4.49(s,2H),4.07(s,2H),3.59(s,1H),2.52-2.51(m,3H).ESI-MS:[M+H] + =664.0.

[0309] Example 19

[0310] Step 1:

[0311] In a 100 mL single-necked bottle, 5-(3-amino-2-fluorobenzyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoic acid hydrochloride (400 mg), 1-hydroxybenzotriazole (104 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (294 mg) and N,N-dimethylformamide (12 mL) were added, and the system was stirred at room temperature for 2 hours. Subsequently, propargylhydroxylamine hydrochloride (92 mg) and N,N-diisopropylethylamine (298 mg) were added to the system, and the reaction was continued to stir at room temperature for 3 hours. The reaction system was added to saturated brine to precipitate a solid. The filter cake was filtered and purified by silica gel column chromatography to obtain 220 mg of a yellow oil. ESI-MS: [M+H] + =570.1.

[0312] Step 2:

[0313] In a 50 mL single-necked bottle, 5-(3-amino-2-fluorobenzyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-N-(prop-2-yn-1-yloxy)benzamide (220 mg), N,N-dimethylformamide (5 mL) and pyridine (493 mg) were added, and the system was placed in an ice bath and stirred. When the system dropped below 0°C, methylaminosulfonyl chloride (505 mg) was slowly added dropwise. After 30 minutes, the reaction was stopped, the system was poured into saturated brine and stirred at room temperature for 10 minutes. Extracted with ethyl acetate, the organic phases were combined and dried over anhydrous sodium sulfate. Filtered and the solvent was removed by rotation, and the crude product was purified by silica gel column chromatography to obtain 140 mg of a light yellow solid. 1 H NMR (400MHz, DMSO-d6): δ11.99(s,1H),9.43(s,1H),8.47(s,1H),8.02(d,J=2 .8Hz,1H),7.57(dd,J1=10.8Hz,J2=2.0Hz,1H),7.36-7.32(m,2H),7.25(q,J=4 .8Hz,1H),7.11(t,J=8.0Hz,1H),7.02(t,J=6.8Hz,1H),6.65-6.62(m,1H),4.4 8(d,J=1.6Hz,2H),4.02(s,2H),3.59(s,1H),2.53-2.51(m,3H).ESI-MS:[M+H] + =663.0.

[0314] Example 20

[0315] Step 1:

[0316] In a 1L three-necked flask, add but-3-yn-1-ol (6.0g), triphenylphosphine (29.2g), N-hydroxyphthalimide (15.4g) and tetrahydrofuran (342mL), and replace the system with a nitrogen atmosphere. Cool the system to below 0°C, and add a tetrahydrofuran solution of diisopropyl azodicarboxylate (22.5g, 111mL tetrahydrofuran) dropwise to the system. After the addition is complete, remove the low-temperature bath and stir the system at room temperature for 16h. Concentrate the reaction system, then add it to saturated brine, extract the combined organic phases with ethyl acetate, dry and concentrate. Purify the concentrate by silica gel column chromatography to obtain 9.2g of an off-white solid.

[0317] Step 2:

[0318] In a 250 mL single-necked flask, add 2-(but-3-yn-1-yloxy)isoindoline-1,3-dione (5.0 g) and dichloromethane / methanol (2 / 1, v / v, 117 mL in total), and replace the system with a nitrogen atmosphere. Slowly add hydrazine hydrate (1.6 g) dropwise to the system. After addition, continue stirring and react for 3 hours. Filter with suction, wash the filter cake with dichloromethane, and combine the organic phases. Concentrate, dilute with tert-methyl ether, filter with suction, and adjust the filtrate pH to 1 with a solution of hydrogen chloride in ethyl acetate. Concentrate the filtrate to obtain 670 mg of a white solid.

[0319] Step 3:

[0320] In a 50 mL single-necked flask, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoic acid hydrochloride (400 mg), 1-hydroxybenzotriazole (125 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (296 mg), and N,N-dimethylformamide (10 mL) were added, and the system was stirred at room temperature for 2 hours. Subsequently, O-(but-3-yn-1-yl)hydroxylamine hydrochloride (103 mg) and N,N-diisopropylethylamine (274 mg) were added to the system, and the reaction was continued to stir at room temperature for 2 hours. The reaction system was added to saturated brine to precipitate a solid. The filter cake was filtered and purified by silica gel column chromatography to obtain 150 mg of a yellow solid. ESI-MS: [M+H] + =585.1.

[0321] Step 4:

[0322] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-N-(but-3-yn-1-yloxy)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzamide (150 mg), N,N-dimethylformamide (5 mL) and pyridine (205 mg) were added, and the system was placed in an ice bath and stirred. When the system temperature dropped below 0°C, methylaminosulfonyl chloride (100 mg) was slowly added dropwise. The reaction was stopped after 30 minutes, and the system was poured into a saturated aqueous sodium bicarbonate solution and stirred at room temperature for 10 minutes. Extracted with ethyl acetate, the organic phases were combined and dried over anhydrous sodium sulfate. Filtered and the solvent was removed by rotation, and the crude product was purified by silica gel column chromatography to obtain 100 mg of an off-white solid. 1 H NMR (400MHz, DMSO-d6): δ11.86(s,1H),10.42(s,1H),8.54(s,1H),8.02(d,J=5.2Hz ,1H),7.57(dd,J1=10.8Hz,J2=1.6Hz,1H),7.41(d,J=7.2Hz,1H),7.35(d,J=8.0Hz,1 H),7.03(q,J=4.8Hz,1H),6.94(t,J=5.2Hz,1H),6.68-6.63(m,1H),4.07(s,2H),3.8 5(t,J=6.4Hz,2H),2.86(s,1H),2.52-2.51(m,3H),2.49-2.47(m,2H).ESI-MS:[M+H] + =678.0.

[0323] Example 21

[0324] Step 1:

[0325] In a 1L three-necked flask, pent-4-yn-1-ol (6.0g), triphenylphosphine (28.1g), N-hydroxyphthalimide (15.0g) and tetrahydrofuran (300mL) were added, and the system was replaced with a nitrogen environment. The system was cooled to below 0°C, and a tetrahydrofuran solution of diisopropyl azodicarboxylate (21.6g, 107mL tetrahydrofuran) was added dropwise to the system. After the dropwise addition, the low-temperature bath was removed and the system was stirred at room temperature for 16h. The reaction system was concentrated and then added to saturated brine. The organic phases were extracted with ethyl acetate, dried and concentrated. Purification by silica gel column chromatography gave 9.1g of an off-white solid.

[0326] Step 2:

[0327] In a 250 mL single-necked flask, add 2-(pent-4-yn-1-yloxy)isoindoline-1,3-dione (8.0 g:) and dichloromethane / methanol (2 / 1, v / v, 175 mL in total), and replace the system with a nitrogen atmosphere. Slowly add hydrazine hydrate (2.4 g) dropwise to the system. After addition, continue stirring and react for 3 hours. Filter with suction, wash the filter cake with dichloromethane, and combine the organic phases. Concentrate, dilute the concentrate with tertiary methyl ether, filter with suction, and adjust the filtrate pH to 1 with a solution of hydrogen chloride in ethyl acetate. Concentrate the filtrate to obtain 2.5 g of a light yellow oil.

[0328] Step 3:

[0329] In a 100 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoic acid hydrochloride (300 mg), 1-hydroxybenzotriazole (88 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (207 mg) and N,N-dimethylformamide (5 mL) were added, and the system was stirred at room temperature for 2 hours. Subsequently, O-(pent-4-yn-1-yl)hydroxylamine hydrochloride (146 mg) and N,N-diisopropylethylamine (209 mg) were added to the system, and the reaction was continued to stir at room temperature for 2 hours. The reaction system was added to saturated brine to precipitate a solid. Filter and dry the filter cake to obtain 300 mg of a yellow oil. ESI-MS: [M+H] + =599.1.

[0330] Step 4:

[0331] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-N-(pent-4-yn-1-yloxy)benzamide (300 mg), N,N-dimethylformamide (5 mL) and pyridine (390 mg) were added, and the system was placed in an ice bath and stirred. When the system dropped below 0 ° C, methylaminosulfonyl chloride (194 mg) was slowly added dropwise. After 30 minutes, the reaction was stopped, the system was poured into saturated brine and stirred at room temperature for 10 minutes. Extracted with ethyl acetate, the organic phases were combined and dried over anhydrous sodium sulfate. Filtered and the solvent was removed by rotation, and the crude product was purified by silica gel column chromatography to obtain 210 mg of a yellow solid. 1H NMR (400MHz, DMSO-d6): δ11.76(s,1H),10.42(s, 1H),8.53(s,1H),8.02(d,J=5.2Hz,1H),7.57(dd,J1=10.8Hz,J2=1.6Hz,1H),7 .38(d,J=7.2Hz,1H),7.35(d,J=8.8Hz,1H),7.04(q,J=5.2Hz,1H),6.94(t,J=4. 8Hz,1H),6.68-6.63(m,1H),4.08(s,2H),3.83(t,J=6.0Hz,2H),2.80(t,J=2.4 Hz,1H),2.52-2.51(m,3H),2.28-2.26(m,2H),1.74-1.68(m,2H).ESI-MS:[M+H] + =692.0.

[0332] Example 22

[0333] Step 1:

[0334] In a 500mL three-necked flask, add but-2-yn-1-ol (2.0g), triphenylphosphine (9.7g), N-hydroxyphthalimide (5.1g) and tetrahydrofuran (114mL), and replace the system with a nitrogen atmosphere. The system is cooled to below 0°C, and a tetrahydrofuran solution of diisopropyl azodicarboxylate (7.0g, 37mL tetrahydrofuran) is added dropwise to the system. After the addition is complete, remove the low-temperature bath and stir the system at room temperature for 16h. Concentrate the reaction system, then add it to saturated brine, extract with ethyl acetate, combine the organic phases, dry and concentrate. Purify the concentrate by silica gel column chromatography to obtain 3.1g of an off-white solid.

[0335] Step 2:

[0336] In a 250 mL single-necked flask, add 2-(but-2-yn-1-yloxy)isoindoline-1,3-dione (4.6 g) and dichloromethane / methanol (1 / 1, v / v, 100 mL total), and replace the system with a nitrogen atmosphere. Slowly add hydrazine hydrate (1.5 g) dropwise to the system. After addition, continue stirring and react for 3 hours. Filter with suction, wash the filter cake with dichloromethane, and combine the organic phases. Concentrate, dilute the concentrate with tertiary methyl ether, filter with suction, and adjust the filtrate pH to 1 with a solution of hydrogen chloride in ethyl acetate. Concentrate the filtrate to obtain 2.1 g of a light yellow oil.

[0337] Step 3:

[0338] In a 100 mL single-necked flask, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoic acid hydrochloride (400 mg), 1-hydroxybenzotriazole (117 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (276 mg), and N,N-dimethylformamide (5 mL) were added, and the system was stirred at room temperature for 2 hours. Subsequently, O-(but-2-yn-1-yl)hydroxylamine hydrochloride (123 mg) and N,N-diisopropylethylamine (186 mg) were added to the system, and the reaction was continued to stir at room temperature for 2 hours. The reaction system was added to saturated brine to precipitate a solid. The filter cake was filtered and purified by silica gel column chromatography to obtain 250 mg of a yellow solid. ESI-MS: [M+H] + =585.1.

[0339] Step 4:

[0340] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-N-(but-2-yn-1-yloxy)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzamide (250 mg), N,N-dimethylformamide (5 mL) and pyridine (336 mg) were added, and the system was placed in an ice bath and stirred. When the system dropped below 0°C, methylaminosulfonyl chloride (167 mg) was slowly added dropwise. After 30 minutes, the reaction was stopped, the system was poured into saturated brine and stirred at room temperature for 10 minutes. Extracted with ethyl acetate, the organic phases were combined and dried over anhydrous sodium sulfate. Filtered and the solvent was removed by rotation, and the crude product was purified by silica gel column chromatography to obtain 180 mg of an off-white solid. 1 H NMR (400MHz, DMSO-d6): δ11.94(s,1H),10.42(s,1H),8.52(s,1H),8.03(d,J=5.2Hz,1H),7.58(dd,J1=10.8Hz,J2=1.6Hz,1H),7.36-7.35(m, 2H),7.03(q,J=4.8Hz,1H),6.95(t,J=4.8Hz,1H),6.70-6.64(m,1H),4.43(s,2H),4.07(s,2H),2.52-2.51(m,3H),1.80(s,3H).ESI-MS:[M+H] + =678.0.

[0341] Example 23

[0342] Step 1:

[0343] In a 500mL three-necked flask, 3-cyclopropylprop-2-yn-1-ol (4.0g), triphenylphosphine (14.2g), N-hydroxyphthalimide (7.5g) and tetrahydrofuran (166mL) were added, and the system was replaced with a nitrogen environment and stirred at room temperature for reaction. The system was cooled to below 0°C, and a tetrahydrofuran solution of diisopropyl azodicarboxylate (10.9g, 54mL tetrahydrofuran) was added dropwise to the system. After the dropwise addition, the low temperature bath was removed and the system was stirred at room temperature for 16h. The reaction system was concentrated and then added to saturated brine, extracted with ethyl acetate, and the organic phases were combined, dried and concentrated. The concentrate was purified by silica gel column chromatography to obtain 6.7g of an off-white solid.

[0344] Step 2:

[0345] In a 250 mL single-necked flask, add 2-((3-cyclopropylprop-2-yn-1-yl)oxy)isoindoline-1,3-dione (6.7 g) and dichloromethane / methanol (2 / 1, v / v, 162 mL in total), and replace the system with a nitrogen atmosphere. Slowly add hydrazine hydrate (1.9 g) dropwise to the system. After addition, continue stirring and react for 3 hours. Filter with suction, wash the filter cake with dichloromethane, and combine the organic phases. Concentrate, dilute the concentrate with tertiary methyl ether, filter with suction, and adjust the filtrate pH to 1 with a solution of hydrogen chloride in ethyl acetate. Concentrate the filtrate to obtain 2.0 g of a light yellow oil.

[0346] Step 3:

[0347] In a 100 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoic acid hydrochloride (220 mg), 1-hydroxybenzotriazole (81 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (230 mg) and N,N-dimethylformamide (10 mL) were added, and the system was stirred at room temperature for 2 hours. Subsequently, O-(3-cyclopropylprop-2-yn-1-yl)hydroxylamine hydrochloride (118 mg) and N,N-diisopropylethylamine (268 mg) were added to the system, and the reaction was continued to stir at room temperature for 2 hours. The reaction system was added to saturated brine to precipitate a solid. The filter cake was filtered and concentrated to obtain 150 mg of a yellow solid. ESI-MS: [M+H] + =611.1.

[0348] Step 4:

[0349] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-N-((3-cyclopropylprop-2-yn-1-yl)oxy)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzamide (235 mg), N,N-dimethylformamide (10 mL) and pyridine (314 mg) were added, and the system was placed in an ice bath and stirred. When the system dropped below 0°C, methylaminosulfonyl chloride (311 mg) was slowly added dropwise. After the addition was completed, the reaction was continued for 30 minutes and then stopped. The system was poured into saturated brine and stirred at room temperature for 10 minutes. Extracted with ethyl acetate, the organic phases were combined and dried over anhydrous sodium sulfate. Filtered and the solvent was removed by rotation, and the crude product was purified by silica gel column chromatography to obtain 10 mg of a yellow solid. 1 H NMR (400MHz, DMSO-d6): δ11.92(s,1H),10.42(s,1H),8.54(s,1H),8.02(d, J=4.8Hz,1H),7.58(d,J=10.8Hz,1H),7.36-7.35(m,2H),7.04(d,J=4.8Hz,1 H),6.94(s,1H),6.70-6.65(m,1H),4.42(s,2H),4.07(s,2H),2.52-2.51(m, 3H),1.31-1.29(m,1H),0.77-0.69(m,2H),0.64-0.58(m,2H).ESI-MS:[M+H] + =704.0.

[0350] Example 24

[0351] To a 50 mL single-necked flask was added N-methyl-d3-2-oxoxooxazoline-3-sulfonamide (641 mg), 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-N-(prop-2-yn-1-yloxy)benzamide (200 mg), triethylamine (354 mg), and 1,2-dichloroethane (10 mL). The reaction system was heated to 80°C for 17 h. After cooling to room temperature, the system was poured into a water / tetrahydrofuran mixture (90 / 10, v / v), and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, dried, concentrated, and purified by silica gel column chromatography to obtain 50 mg of a yellow solid. 1H NMR (400MHz, DMSO-d6): δ12.02(s,1H),10.41(s,1H),8.53(s,1H),8.03(d,J=4.8Hz,1H),7.58(dd,J1=10.8Hz,J2=2.0Hz,1H),7.38-7.3 5(m,2H),7.01(s,1H),6.95(t,J=4.4Hz,1H),6.70-6.65(m,1H),4.90(d,J=2.0Hz,2H),4.07(s,2H),3.59(t,J=2.0Hz,1H).ESI-MS:[M+H] + =667.0.

[0352] Example 25

[0353] To a 50 mL single-necked flask was added N-ethyl-2-oxoxazoline-3-sulfonamide (339 mg), 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-N-(prop-2-yn-1-yloxy)benzamide (100 mg), triethylamine (177 mg), and 1,2-dichloroethane (5 mL). The reaction system was heated to 80°C for 17 h. After cooling to room temperature, the system was poured into a water / tetrahydrofuran mixture (90 / 10, v / v), and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, dried, concentrated, and purified by silica gel column chromatography to obtain 5 mg of an off-white solid. 1 H NMR (400MHz, DMSO-d6): δ12.02(s,1H),10.40(s,1H),8.52(s,1H),8.02(d,J=4.8Hz,1H),7.57(dd,J1=10.4Hz,J2=1.2Hz,1H),7.36(d,J=8.4Hz,2H), 7.18(s,1H),6.93(t,J=4.0Hz,1H),6.97-6.64(m,1H),4.49(s,2H),4.06(s ,2H),3.58(s,1H),2.96-2.91(m,2H),1.02(t,J=7.2Hz,3H).ESI-MS:[M+H] + =678.0.

[0354] Example 26

[0355] Step 1:

[0356] To a 3L reaction flask, add 1000mL of dichloromethane, trimethylsilyl trifluoromethanesulfonate (271.8g), triethylamine (123.7g), and silver trifluoromethanesulfonate (10g). Slowly add a solution of ethyl propargyl acid (100g) in dichloromethane (100mL) dropwise. After the addition is complete, stir at room temperature for 30 minutes and concentrate under reduced pressure to remove the dichloromethane. Extract the concentrated residue with n-hexane. Combine the n-hexanes and wash with saturated ammonium chloride solution. Dry the n-hexane phase over anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain approximately 140g of a light yellow liquid.

[0357] Step 2:

[0358] In a 1000mL three-necked flask, add ethyl 3-(trimethylsilyl)propionate (50g), 500mL of ether, sodium borodeuteride (17.3g), lithium chloride (17.4g), and heavy water (8.2g). Stir at room temperature for 40 minutes, then warm to 33°C and react for 8 hours. Slowly add the reaction solution to a 10% aqueous acetic acid solution; extract with n-hexane, separate the layers, and extract the aqueous layer again with n-hexane. Combine the organic phases, wash with 1N hydrochloric acid solution, and then wash the organic phase with water until neutral. Dry over anhydrous sodium sulfate and concentrate under reduced pressure to obtain a light yellow liquid. 1 H NMR (400MHz, DMSO-d6): δ5.20 (brs, 1H), 0.13 (s, 9H).

[0359] Step 3:

[0360] In a 500 mL three-necked flask, 3-(trimethylsilyl)prop-2-yn-1,1-d2-1-ol (5.0 g), triphenylphosphine (13.1 g), N-hydroxyphthalimide (6.9 g), and tetrahydrofuran (250 mL) were added and the system was replaced with a nitrogen atmosphere. The system was cooled to below 0°C, and a solution of diisopropyl azodicarboxylate in tetrahydrofuran (10.1 g in 100 mL of tetrahydrofuran) was added dropwise. After the addition was complete, the low-temperature bath was removed and the system was stirred at room temperature for 16 hours. The reaction system was concentrated and then added to saturated brine. The organic phases were extracted with ethyl acetate, dried, and concentrated. The concentrate was purified by silica gel column chromatography to obtain 5.5 g of a white solid.

[0361] Step 4:

[0362] In a 250 mL single-necked bottle, 2-((3-(trimethylsilyl)prop-2-yn-1-yl-1,1-d2)oxy)isoindoline-1,3-dione (1.0 g) and dichloromethane / methanol (2 / 1, v / v, 24 mL in total) were added, and the system was replaced with a nitrogen atmosphere. Hydrazine hydrate (218 mg) was slowly added dropwise to the system. After the addition, the reaction was stirred for 3 h. Filtered with suction, the filter cake was washed with dichloromethane, and the organic phases were combined. Concentrated, the concentrate was diluted with tert-methyl ether, filtered, and the pH of the filtrate was adjusted to 1 with a solution of hydrogen chloride in ethyl acetate. The filtrate was concentrated to obtain 500 mg of a light yellow oil.

[0363] Step 5:

[0364] In a 100 mL single-necked flask, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoic acid hydrochloride (500 mg), 1-hydroxybenzotriazole (146 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (345 mg), and N,N-dimethylformamide (10 mL) were added, and the system was stirred at room temperature for 2 hours. Subsequently, O-(3-(trimethylsilyl)prop-2-yn-1-yl-1,1-d2)hydroxylamine hydrochloride (262 mg) and N,N-diisopropylethylamine (232 mg) were added to the system, and the reaction was continued to stir at room temperature for 2 hours. The reaction system was added to saturated brine, and a solid precipitated. The filter cake was filtered and purified by silica gel column chromatography to obtain 400 mg of a light yellow solid. ESI-MS: [M+H] + =645.1.

[0365] Step 6:

[0366] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-N-((3-(trimethylsilyl)prop-2-yn-1-yl-1,1-d2)oxy)benzamide (400 mg), tetrahydrofuran (10 mL) and tetrabutylammonium fluoride trihydrate (489 mg) were added and stirred at room temperature for 10 minutes. The system was poured into water, extracted with ethyl acetate, and the organic phases were combined and dried over anhydrous sodium sulfate. Concentration gave 350 mg of a yellow solid. ESI-MS: [M+H] + =573.0.

[0367] Step 7:

[0368] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-N-((prop-2-yn-1-yl-1,1-d2)oxy)benzamide (350 mg), N,N-dimethylformamide (10 mL) and pyridine (476 mg) were added, and the system was placed in an ice bath and stirred. After the system dropped below 0°C, methylaminosulfonyl chloride (237 mg) was slowly added dropwise. After the addition was completed, the reaction was continued for 30 minutes and then stopped. The system was poured into a saturated aqueous sodium bicarbonate solution and stirred at room temperature for 10 minutes. Extracted with ethyl acetate, the organic phases were combined and dried over anhydrous sodium sulfate. Filtered and the solvent was removed by rotation, and the crude product was purified by silica gel column chromatography to obtain 200 mg of an off-white solid. 1 H NMR (400MHz, DMSO-d6): δ12.02(s,1H),10.42(s,1H),8.53(s,1H),8.03(d,J=4.8Hz,1H),7.58(dd,J1=10.8Hz,J2=1.6Hz,1H),7.37-7. 35(m,2H),7.04(d,J=5.2Hz,1H),6.95(t,J=4.8Hz,1H),6.70-6.65(m,1H),4.07(s,2H),3.58(s,1H),2.52-2.51(m,3H).ESI-MS:[M+H] + =666.0.

[0369] Example 27

[0370] Step 1:

[0371] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-methylphenyl)amino)benzoic acid hydrochloride (500 mg), 1-hydroxybenzotriazole (375 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (472 mg) and N,N-dimethylformamide (5 mL) were added, and the system was stirred at room temperature for 2 hours. Subsequently, propargylhydroxylamine hydrochloride (397 mg) and N,N-diisopropylethylamine (952 mg) were added to the system, and the reaction was continued to stir at room temperature for 2 hours. The reaction system was added to saturated brine to precipitate a solid. The filter cake was filtered and purified by silica gel column chromatography to obtain 250 mg of a yellow solid. ESI-MS: [M+H] + =459.3.

[0372] Step 2:

[0373] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-methylphenyl)amino)-N-(prop-2-yn-1-yloxy)benzamide (350 mg), N,N-dimethylformamide (5 mL) and pyridine (594 mg) were added, and the system was placed in an ice bath and stirred. When the system dropped below 0°C, methylaminosulfonyl chloride (295 mg) was slowly added dropwise. After 30 minutes, the reaction was stopped, the system was poured into saturated brine and stirred at room temperature for 10 minutes. Extracted with ethyl acetate, the organic phases were combined and dried over anhydrous sodium sulfate. Filtered and the solvent was removed by rotation, and the crude product was purified by silica gel column chromatography to obtain 180 mg of an off-white solid. 1 H NMR (400MHz, DMSO-d6): δ12.06(s,1H),10.42(s,1H),8.54(s,1H),8.02(d,J=4.8Hz,1H),7.36(d,J=6.8Hz,1H),7.05-7.02(m,2H ),6.94(t,J=4.4Hz,1H),6.88-6.80(m,2H),4.51(s,2H),4.04(s,2H),3.60(s,1H),2.53-2.51(m,3H),2.25(s,3H).ESI-MS:[M+H] + =552.3.

[0374] Example 28

[0375] Step 1:

[0376] In a 50 mL single-necked bottle, methyl 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoate (1.3 g), potassium vinyl fluoroborate (636 mg), potassium carbonate (984 mg), 1,4-dioxane (12 mL), water (4 mL) and [1,1′-bis(diphenylphosphino)ferrocene]palladium dichloride (86 mg) were added, and the system was replaced with a nitrogen atmosphere and heated to 95 ° C. The reaction was stirred for 2 hours. After the system was cooled to room temperature, it was poured into water to quench the reaction. It was extracted with ethyl acetate, the organic phases were combined, and dried over anhydrous sodium sulfate. Filtered and the solvent was removed by rotation to obtain 950 mg of a brown solid. ESI-MS: [M+H] + =432.2.

[0377] Step 2:

[0378] In a 50 mL single-necked flask, methyl 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-vinylphenyl)amino)benzoate (950 mg), tetrahydrofuran (20 mL), and water (10 mL) were added and stirred at room temperature until the solution became clear. The system was then placed in an ice bath and lithium hydroxide monohydrate (360 mg) was slowly added to the system. After the addition, the system was warmed to room temperature and stirred for 3 hours. Hydrochloric acid (6 M, 5 mL) was then slowly added dropwise to the system to adjust the pH to 1-2 and stirred for 5 minutes. The tetrahydrofuran in the system was removed under reduced pressure, and then water was added to the system to dilute the concentrate and stir for 10 minutes. The filter cake was collected by filtration and washed with water and tertiary methyl ether to obtain 700 mg of a brown solid. The product from this step was used directly in the next step without further characterization or purification.

[0379] Step 3:

[0380] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-vinylphenyl)amino)benzoic acid hydrochloride (300 mg), 1-hydroxybenzotriazole (107 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (253 mg) and N,N-dimethylformamide (7 mL) were added, and the system was stirred at room temperature for 2 hours. Subsequently, propargylhydroxylamine hydrochloride (142 mg) and N,N-diisopropylethylamine (255 mg) were added to the system, and the reaction was continued to stir at room temperature for 2 hours. The reaction system was added to saturated brine to precipitate a solid. The filter cake was filtered and purified by silica gel column chromatography to obtain 250 mg of a yellow solid. ESI-MS: [M+H] + =471.2.

[0381] Step 4:

[0382] In a 50 mL single-necked flask, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-vinylphenyl)amino)-N-(prop-2-yn-1-yloxy)benzamide (250 mg), N,N-dimethylformamide (5 mL), and pyridine (414 mg) were added, and the system was placed in an ice bath and stirred. After the system cooled to below 0°C, methylaminosulfonyl chloride (206 mg) was slowly added dropwise. The reaction was stopped after 30 minutes, and the system was poured into saturated brine and stirred at room temperature for 10 minutes. Extraction was performed with ethyl acetate, and the organic phases were combined and dried over anhydrous sodium sulfate. Filter and spin-dry the solvent. The crude product was purified by silica gel column chromatography to obtain 120 mg of an off-white solid. 1H NMR (400MHz, DMSO-d6): δ12.05(s,1H),10.42(s,1H),8.60(s,1H),8.03(d,J=5.2Hz,1H),7. 38(d,J=10.8Hz,2H),7.13(d,J=8.4Hz,1H),7.04(q,J=4.8Hz,1H),6.95(d,J=4.8Hz,1H),6. 87-6.82(m,1H),6.64(dd,J1=17.6Hz,J2=11.2Hz,1H),5.74(d,J=17.2Hz,1H),5.18(d,J=10 .8Hz,1H),4.50(d,J=1.6Hz,2H),4.07(s,2H),3.59(s,1H),2.53-2.51(m,3H).ESI-MS:[M+H] + =564.2.

[0383] Example 29

[0384] Step 1:

[0385] In a 100mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoic acid methyl ester (1.5g), cuprous iodide (11mg), triethylamine (4.5mL), tetrahydrofuran (15mL) and bistriphenylphosphine palladium dichloride (40mg) were added, and the system was replaced with a nitrogen environment and stirred at room temperature for reaction. Trimethylsilyl acetylene (1.1g) was then slowly added dropwise to the system. After the addition, the reaction system was stirred at room temperature for 21h. The reaction system was diluted with ethyl acetate, filtered, the filtrate was collected, washed with saturated brine, the organic phase was collected, and dried over anhydrous sodium sulfate. Filter and spin to remove the solvent, and the crude product was purified by silica gel column chromatography to obtain 1.0g of an off-white solid. ESI-MS: [M+H] + =502.2.

[0386] Step 2:

[0387] In a 50 mL single-necked bottle, methyl 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-((trimethylsilyl)ethynyl)phenyl)amino)benzoate (1.0 g), tetrahydrofuran (15 mL) and water (7.5 mL) were added, and the system was stirred at room temperature until clear. The system was then placed in an ice bath, and lithium hydroxide monohydrate (252 mg) was slowly added to the system. After the addition, the system was warmed to room temperature and stirred for 16.5 hours. Hydrochloric acid (6 M, 6 mL) was then slowly added dropwise to the system to adjust the pH of the system to 1-2 and stirred for 5 minutes. The tetrahydrofuran in the system was removed under reduced pressure, and then the concentrate was diluted with water and stirred for 10 minutes. The filter cake was collected by filtration and washed once with water and tertiary methyl ether to obtain 810 mg of a brown solid. ESI-MS: [M+H] + =416.1.

[0388] Step 3:

[0389] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-2-((4-ethynyl-2-fluorophenyl)amino)-3,4-difluorobenzoic acid hydrochloride (415 mg), 1-hydroxybenzotriazole (135 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (383 mg) and N,N-dimethylformamide (10 mL) were added, and the system was stirred at room temperature for 2 hours. Subsequently, propargylhydroxylamine hydrochloride (215 mg) and N,N-diisopropylethylamine (516 mg) were added to the system, and the reaction was continued to stir at room temperature for 2 hours. The reaction system was added to saturated brine to precipitate a solid. The filter cake was filtered and dried to obtain 300 mg of a brown solid. ESI-MS: [M+H] + =469.1.

[0390] Step 4:

[0391] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-2-((4-ethynyl-2-fluorophenyl)amino)-3,4-difluoro-N-(prop-2-yn-1-yloxy)benzamide (300 mg), N,N-dimethylformamide (5 mL) and pyridine (480 mg) were added, and the system was placed in an ice bath and stirred. When the system dropped below 0°C, methylaminosulfonyl chloride (450 mg) was slowly added dropwise. After 30 minutes, the reaction was stopped, the system was poured into saturated brine and stirred at room temperature for 10 minutes. Extracted with ethyl acetate, the organic phases were combined and dried over anhydrous sodium sulfate. Filtered and the solvent was removed by rotation, and the crude product was purified by silica gel column chromatography to obtain 39 mg of an off-white solid. 1H NMR (400MHz, DMSO-d6): δ12.03(s,1H),10.43(s,1H),8.62(s,1H),8.03(s,1H),7.37-7.32(m,2H) ,7.16-6.79(m,4H),4.49(s,2H),4.12-4.02(m,3H),3.58(s,1H),2.53-2.51(m,3H).ESI-MS:[M+H] + =562.1.

[0392] Example 30

[0393] Step 1:

[0394] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-methoxyphenyl)amino)benzoic acid hydrochloride (250 mg), 1-hydroxybenzotriazole (89 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (211 mg) and N,N-dimethylformamide (5 mL) were added, and the system was stirred at room temperature for 2 hours. Subsequently, propargylhydroxylamine hydrochloride (118 mg) and N,N-diisopropylethylamine (355 mg) were added to the system, and the reaction was continued to stir at room temperature for 2 hours. The reaction system was added to saturated brine to precipitate a solid. Filter and dry the filter cake to obtain 250 mg of a yellow solid. ESI-MS: [M+H] + =475.2.

[0395] Step 2:

[0396] In a 50 mL single-necked bottle, 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-methoxyphenyl)amino)-N-(prop-2-yn-1-yloxy)benzamide (250 mg), N,N-dimethylformamide (5 mL) and pyridine (414 mg) were added, and the system was placed in an ice bath and stirred. When the system dropped below 0°C, methylaminosulfonyl chloride (206 mg) was slowly added dropwise. After 30 minutes, the reaction was stopped, the system was poured into saturated brine and stirred at room temperature for 10 minutes. Extracted with ethyl acetate, the organic phases were combined and dried over anhydrous sodium sulfate. Filtered and the solvent was removed by rotation, and the crude product was purified by silica gel column chromatography to obtain 80 mg of a yellow solid. 1H NMR (400MHz, DMSO-d6): δ12.04(s,1H),10.41(s,1H),8.53(s,1H),8.02(d,J=4 .8Hz,1H),7.34(d,J=6.8Hz,1H),7.06-6.95(m,2H),6.92(t,J=4.8Hz,1H),6.8 7(dd,J1=13.2Hz,J2=2.8Hz,1H),6.69-6.66(m,1H),4.51(d,J=2.0Hz,2H),4.0 1(s,2H),3.73(s,3H),3.61(t,J=2.0Hz,1H),2.52-2.51(m,3H).ESI-MS:[M+H] + =568.1.

[0397] Preparation of control compounds

[0398] The reference compounds Ref A-7 and Ref A-13 were synthesized with reference to CN114929669, which are compound A-7 and compound A-13 in the reference, respectively, and have the following structures:

[0399] The therapeutic efficacy and safety of a drug in vivo are the result of the combined effects of the compound's inhibitory activity, various physicochemical properties, and absorption, metabolism, distribution, excretion, and other factors. The compounds of this invention demonstrate superior efficacy and safety and have promising therapeutic prospects.

[0400] Biological testing

[0401] 1. Determination of Cell Proliferation Inhibitory Activity

[0402] Cells in the exponential growth phase were digested with trypsin-EDTA, and HCT-116 cells were plated in 96-well plates and cultured overnight at 37°C, 5% CO2. Growth medium containing the compound was added, and the cells were incubated in an incubator at 37°C, 5% CO2. After incubation, a luminescence detection reagent was added to each well, and the cells were shaken and incubated. The chemiluminescence value was measured, and the IC value of the compound for inhibiting HCT-116 cell proliferation was fitted and calculated. 50 value.

[0403] Where "A" stands for IC 50 Value (nM) ≤ 150;

[0404] "B" stands for IC 50 Value (nM) 150~300 (excluding 300);

[0405] "C" stands for IC 50 Value (nM) 300~500 (excluding 500);

[0406] "D" stands for IC 50 Value (nM) ≥ 500;

[0407] Compound 1 of the present invention exhibits significant anti-proliferative activity against HCT-116 cells.

[0408] 2. BRAF Inhibitory Activity Test

[0409] Fluorescence polarization testing was performed using the following procedure: a series of test substances were added to a mixture of BRAF and MAP2K1 fluorescent peptides, mixed, and centrifuged; after incubation at 23°C, ATP buffer was added, mixed again, centrifuged, and incubated; finally, the detection reagent was added, mixed, centrifuged, and incubated, and polarized fluorescence was measured using a Perkin Elmer Envision. Data were fitted using XLFIT5 software, and the results are shown in the table below:

[0410] In the above table, "A" represents the inhibitory activity IC 50 <5nM, "B" indicates inhibitory activity 5nM ≤ IC 50 <10nM, "C" indicates inhibitory activity 10nM ≤ IC 50 <50nM.

[0411] The test results showed that the compound of the present invention exhibited significant inhibitory activity against BRAF.

[0412] 3. MEK1 inhibitory activity test

[0413] Fluorescence polarization testing was performed using the following procedure: MEK1 enzyme solution or control buffer was mixed with a series of test substances at varying concentrations, centrifuged, and incubated at 23°C. Inactive ERK, BRAF, ATP, and a fluorescent peptide substrate were then added, mixed, centrifuged, and incubated. Finally, the detection reagent was added, mixed, centrifuged, and incubated, and polarized fluorescence was measured using a Perkin Elmer Envision. Data were fitted using XLFIT5 software, and the results are shown in the table below:

[0414] In the above table, "A" represents the inhibitory activity IC 50 <5nM, "B" indicates inhibitory activity 5nM ≤ IC 50 <10nM, "C" indicates inhibitory activity 10nM ≤ IC 50 <50nM.

[0415] The test results showed that the compound of the present invention exhibited significant inhibitory activity against MEK1.

[0416] 4. Evaluation of the Antiproliferation Activity of Compounds on Tumor Cells SW480

[0417] The CellTiter-3D-Glo reagent method was used to detect the proliferation inhibitory activity of the 3D model of colorectal cancer cell SW480. The experimental process is as follows: SW480 cells in the logarithmic growth phase were digested and seeded into low-adsorption 96-well U-bottom plates at the corresponding density and cultured in a cell culture incubator; culture medium containing a series of concentration gradient test substances was added to the experimental wells, and control medium was added to the control wells, and incubated in a 5% CO2 incubator. After incubation, celltiter-3D-glo reagent was added to each well for lysis detection; chemiluminescence values ​​were measured by microplate reader. Data were fitted using Graphpad Prism software to calculate the IC value of the test substance. 50 The test results are shown in the following table:

[0418] The test results showed that the compound of the present invention exhibited significant cell proliferation inhibitory activity on colorectal cancer cell line SW480.

[0419] 5. Evaluation of the compound's proliferation inhibitory activity against tumor cell A549

[0420] The CellTiter-3D-Glo reagent method was used to detect the proliferation inhibitory activity of the 3D model of lung cancer A549 cells. The experimental process is as follows: A549 cells in the logarithmic growth phase were digested and seeded into low-adsorption 96-well U-bottom plates at the corresponding density and cultured in a cell culture incubator; culture medium containing a series of concentration gradients of the test substance was added to the experimental wells, and control medium was added to the control wells, and incubated in a 5% CO2 incubator. After incubation, the celltiter-3D-glo reagent was added to each well for lysis detection; the chemiluminescence value was measured by microplate reader. The data were fitted with Graphpad Prism software to calculate the IC value of the test substance. 50 The test results are shown in the following table:

[0421] The test results showed that the compound of the present invention exhibited significant cell proliferation inhibition activity on lung cancer cell A549.

[0422] 6. Evaluation of the compound's proliferation inhibitory activity against tumor cells NCI-H358

[0423] The CellTiter-3D-Glo reagent method was used to detect the proliferation inhibitory activity of the 3D model of lung cancer cells NCI-H358. The experimental process is as follows: NCI-H358 cells in the logarithmic growth phase were digested and seeded into low-adsorption 96-well U-bottom plates at the corresponding density and cultured in a cell culture incubator; culture medium containing a series of concentration gradients of the test substance was added to the experimental wells, and control medium was added to the control wells, and incubated in a 5% CO2 incubator. After incubation, the celltiter-3D-glo reagent was added to each well for lysis detection; the chemiluminescence value was measured by microplate reader. The data were fitted using Graphpad Prism software to calculate the IC value of the test substance. 50 The test results are shown in the following table:

[0424] The test results showed that the compound of the present invention exhibited significant cell proliferation inhibition activity on lung cancer cells NCI-H358.

[0425] 7. Pharmacodynamic Evaluation of the Colorectal Cancer Cell HCT116 Subcutaneous Xenograft Model in Nude Mice

[0426] Test samples: Compound 1 and Compound 21;

[0427] Animal species and number: BALB / c nude mice, 6 per group (half male and half female);

[0428] Dosage: oral gavage, once a day;

[0429] Animal grouping and dosage: vehicle blank group, compound 1 group (0.2 mg / kg, QD×22 days), compound 21 group (0.2 mg / kg, QD×22 days);

[0430] Animal model establishment: HCT116 colorectal cancer cells in the logarithmic growth phase were cultured in vitro and collected, and subcutaneously inoculated into the right back of mice. 3 Then, the animals in each group were randomly divided into groups at about 1 hour. Subsequently, the animals in each group were given the drug, and the day of the first drug administration was the first day of the experiment;

[0431] General status observation: observe the animal's administration site, external signs, mental state, general behavior, death, and other abnormal manifestations once a day;

[0432] Tumor volume calculation: V = 0.5 × long diameter × short diameter 2 The tumor growth inhibition rate (TGI) was used to evaluate the inhibitory effect of the compound on tumor growth.

[0433] In the above table, “+” indicates TGI < 70%; “++” indicates TGI is 70% to 100%; and “+++” indicates TGI > 100%.

[0434] The experimental results showed that compounds 1 and 21 had a significant inhibitory effect on the growth of subcutaneous transplanted tumors of colorectal cancer HCT116 cells in nude mice; during the administration period, the general observations of animals in each administration group were normal, showing good safety.

[0435] 8. In vivo pharmacodynamic evaluation of the compound in combination with a KRAS G12C inhibitor

[0436] Test samples: Compound 21, AMG510 (Sotorasib);

[0437] Animal species: BALB / c nude mice, 6 mice / group (half male and half female);

[0438] Dosage: oral gavage, once a day;

[0439] Animal grouping and dosage: vehicle blank group, compound 21 (0.1 mg / kg, QD×22 days), compound 21+AMG510 (0.1+10 mg / kg, QD×22 days), AMG510 (10 mg / kg, QD×22 days);

[0440] Animal model establishment: MIA PaCa-2 lung cancer cells in the logarithmic growth phase were cultured in vitro and collected, and subcutaneously inoculated into the right back of mice. 3 Then, the animals in each group were randomly divided into groups at about 1 hour. Subsequently, the animals in each group were given the drug, and the day of the first drug administration was the first day of the experiment;

[0441] General status observation: observe the animal's administration site, external signs, mental state, general behavior, death, and other abnormal manifestations once a day;

[0442] Tumor volume calculation: V = 0.5 × long diameter × short diameter 2 The tumor growth inhibition rate (TGI) was used to evaluate the inhibitory effect of the compound on tumor growth.

[0443] In the above table, “+” indicates TGI < 70%; “++” indicates TGI is 70% to 100%; and “+++” indicates TGI > 100%.

[0444] The experimental results showed that compound 21 and AMG510, when administered alone, inhibited the growth of subcutaneous xenograft tumors in nude mice, with compound 21 being more effective than AMG510. The combination group showed a significant inhibitory effect on tumor growth compared to the single-dose groups. During the dosing period, the animals in each treatment group maintained normal diet and activity, demonstrating a good safety profile.

Claims

1. A compound represented by formula (I), its stereoisomers, tautomers, solvates and pharmaceutically acceptable salts, in, U1, U2, U3, U4 are each independently selected from nitrogen, CR U ; R U Selected from R 5c , R 5d , R 6b , R 6c ; W1, W2, and W3 are each independently a bond, NR W ; R W is selected from hydrogen, alkyl, deuterated alkyl; Y is selected from oxygen or sulfur; R1 is selected from -S(O)2NH-R7 or -S(O)2-R7; R7 is selected from hydrogen, substituted or unsubstituted alkyl, deuterated alkyl, haloalkyl, alkoxyalkyl, alkoxy, cycloalkyl, alkenyl, alkynyl; R2 is selected from hydrogen, substituted or unsubstituted alkyl, deuterated alkyl, haloalkyl, alkoxy, alkoxyalkyl, alkenyl, alkynyl, cycloalkyl; R 3a , R 3b , R 4a , R 4b , R 4c , R 4d , R 4e , R 5a , R 5b , R 5c , R 5d , R 6a , R 6b , R 6c Each is independently selected from hydrogen, deuterium, halogen, hydroxyl, substituted or unsubstituted amino, alkyl, deuterated alkyl, haloalkyl, alkoxy, alkenyl, alkynyl, alkylthio, cycloalkyl, heterocyclic group, aryl, heteroaryl, or two adjacent substituents can together form a substituted or unsubstituted cyclic structure.

2. A compound represented by formula (II), its stereoisomers, tautomers, solvates and pharmaceutically acceptable salts, in, U1 is selected from nitrogen, CR 5d ; R7 is selected from hydrogen, substituted or unsubstituted alkyl, deuterated alkyl, haloalkyl, alkoxyalkyl, alkoxy, cycloalkyl, alkenyl, alkynyl; R2 is selected from hydrogen, substituted or unsubstituted alkyl, deuterated alkyl, haloalkyl, alkoxy, alkoxyalkyl, alkenyl, alkynyl, cycloalkyl; R 3a , R 3b , R 4a , R 4b , R 4c , R 4d , R 4e , R 5a , R 5b , R 5c , R 5d , R 6a , R 6b , R 6c Each is independently selected from hydrogen, deuterium, halogen, hydroxyl, substituted or unsubstituted amino, alkyl, deuterated alkyl, haloalkyl, alkoxy, alkenyl, alkynyl, alkylthio, cycloalkyl, heterocyclic group, aryl, heteroaryl, or two adjacent substituents can together form a substituted or unsubstituted cyclic structure.

3. A compound represented by formula (III), its stereoisomers, tautomers, solvates and pharmaceutically acceptable salts, in, U1 is selected from nitrogen, CR 5d ; R7 is selected from hydrogen, substituted or unsubstituted alkyl, deuterated alkyl, haloalkyl, alkoxyalkyl, alkoxy, cycloalkyl, alkenyl, alkynyl; R 8a , R 8b , R 8c Each is independently selected from hydrogen, deuterium, halogen, hydroxy, substituted or unsubstituted alkyl, deuterated alkyl, haloalkyl, cycloalkyl, heterocyclyl; R 3a , R 3b , R 4a , R 4b , R 4c , R 4d , R 4e , R 5a , R 5b , R 5c , R 5d , R 6a , R 6b , R 6c are each independently selected from hydrogen, deuterium, halogen, hydroxy, substituted or unsubstituted amino, alkyl, deuterated alkyl, haloalkyl, alkoxy, alkenyl, alkynyl, alkylthio, cycloalkyl, A heterocyclic group, an aryl group, a heteroaryl group, or two adjacent substituents may together form a substituted or unsubstituted ring structure; n is 1, 2, 3, 4.

4. The following compounds, their stereoisomers, tautomers, solvates and pharmaceutically acceptable salts, 5. A compound represented by formula (IV), its stereoisomers, tautomers or pharmaceutically acceptable salts, in, U1 is selected from nitrogen or CR 5d ; R 5d Selected from hydrogen or C 1-6 alkyl; W is selected from a chemical bond or oxygen; R 4a , R 4b , R 4c , R 4d , R 4e are independently selected from hydrogen, halogen, aldehyde, C 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 Alkenyl, C 2-6 Alkynyl or C 3-6 Cycloalkyl; R 5a , R 5b , R 5c , R 6a , R 6b , R 6c are each independently selected from hydrogen, halogen or C 1-6 alkyl; R7 is selected from hydrogen, C 1-6 Alkyl, C 1-6 Deuterated alkyl or C 3-6 Cycloalkyl; R 8a , R 8b , R 8c are each independently selected from hydrogen, deuterium, C 1-6 Alkyl or C 3-6 Cycloalkyl; n is 1, 2, 3, 4.

6. A compound represented by formula (V), its stereoisomers, tautomers or pharmaceutically acceptable salts, in, U1 is selected from nitrogen or CH; R 4a is selected from hydrogen or halogen; R 4c Selected from hydrogen, halogen, aldehyde, C 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 Alkenyl, C 2-6 Alkynyl or C 3-6 Cycloalkyl; R7 is selected from C 1-6 Alkyl, C 1-6 Deuterated alkyl or C 3-6 Cycloalkyl; R 8a , R 8b are each independently selected from hydrogen, deuterium or C 1-6 alkyl; R 8c Selected from hydrogen or C 1-6 alkyl; n is 1, 2, 3.

7. The following compounds, their stereoisomers, tautomers or pharmaceutically acceptable salts, 8. A compound represented by formula (VI), its stereoisomers, tautomers or pharmaceutically acceptable salts, in, U1 is selected from nitrogen or CH; R 4a is selected from hydrogen or halogen; R 4c Selected from hydrogen, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 Alkenyl or C 2-6 Alkynyl; R7 is selected from C 1-6 Alkyl, C 1-6 Deuterated alkyl or C 3-6 Cycloalkyl; R 8a , R 8b are each independently selected from hydrogen, deuterium or C 1-6 alkyl; R 8c Selected from hydrogen, C 1-6 Alkyl or C 3-6 Cycloalkyl; n is 1, 2, 3.

9. The following compounds, their stereoisomers, tautomers or pharmaceutically acceptable salts, 10. A pharmaceutical composition comprising a therapeutically effective dose of a compound according to any one of claims 1 to 9, or a stereoisomer, tautomer, solvate or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

11. Use of the compound according to any one of claims 1 to 9 or its pharmaceutically acceptable salt or the pharmaceutical composition according to claim 10 in the preparation of a medicament for preventing and / or treating diseases associated with MEK and Raf-mediated cancers, wherein the compound according to any one of claims 1 to 9 or its pharmaceutically acceptable salt or the pharmaceutical composition according to claim 10 is used alone or in combination with other therapeutic methods including immunotherapy to prevent and / or treat diseases associated with MEK and Raf-mediated cancers.

12. Use of the compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 10, in the preparation of a method for preventing and / or treating a cell proliferative disease, wherein the compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 10, is used alone or in combination with other therapeutic methods including immunotherapy to prevent and / or treat a cell proliferative disease.

13. The method according to claim 11, wherein the disease includes but is not limited to renal cancer, liver cancer, breast cancer, pancreatic cancer, prostate cancer, melanoma, leukemia, malignant lymphoma, ovarian cancer, head and neck cancer, lung cancer, colon cancer, bladder cancer, uterine cancer, etc.