Aromatic hydrazide derivative and medical use thereof

Abstract

The present invention belongs to the field of medicinal chemistry, and relates to an aromatic hydrazide derivative and the medical use thereof. Specifically, disclosed in the present invention are an aromatic hydrazide derivative, and a stereoisomer, tautomer or pharmaceutically acceptable salt thereof. Further discloses are a preparation method of the compound and the use of the compound in preventing and / or treating tumor diseases. The compound shows a good application prospect in preventing and / or treating tumors related to MAPK pathways.

Description

Aromatic hydrazide derivatives and their medical uses Technical Field

[0001] The present invention belongs to the field of medicinal chemistry and relates to a class of aromatic hydrazide derivatives, a preparation method thereof and the application of the compounds in the preparation of anti-tumor drugs. Background Art

[0002] Hydrazide is widely present in the molecular structures of physiologically active molecules and drugs. Its unique structure plays a significant role in improving drug performance, attracting widespread interest from researchers both domestically and internationally in drug design and discovery. For example, the following drug molecules all contain hydrazide structural units.

[0003] Mitogen-activated protein kinases (MAPKs) belong to a class of serine / threonine kinases. The extracellular signal-regulated kinase (ERK) Ras / Raf / MEK / ERK signaling pathway is one of the most important cellular signaling pathways, playing a central regulatory role in multiple key aspects of cell proliferation, differentiation, apoptosis, and metabolism. Studies have shown that this signaling pathway is highly conserved in eukaryotic cells. When cells are stimulated by mitogens, conformational changes in Ras trigger the Raf / MEK / ERK cascade. Activated ERK enters the cell and phosphorylates various substrates, including kinases and transcription factors, triggering a series of physiological and biochemical reactions.

[0004] The Ras / Raf / MEK / ERK signaling pathway is one of the most important dysregulated signaling pathways in the development and progression of human tumors, involved in multiple aspects of tumor cell proliferation, apoptosis, invasion, and metastasis. This pathway has become an important therapeutic target, and drugs targeting this pathway can significantly inhibit oncogenic signaling in tumor cells, such as the marketed MEK inhibitors trametinib, cobimetinib, and selumetinib, and the Raf inhibitors sorafenib and dabrafenib. Complicating matters is the complex interplay between signaling pathways during the development and progression of various cancers. Inhibition of a single target may lead to compensatory activation of other target pathways, thereby weakening the anticancer effect of a single-target inhibitor. Combination inhibitors have shown superior therapeutic efficacy in overcoming or delaying drug resistance compared to single-target inhibitors.

[0005] The development of inhibitors targeting the Ras / Raf / MEK / ERK signaling pathway has made some progress, but it is clear that there is still a need to discover and develop new compounds with better efficacy, pharmacokinetic properties and safety. Such research work is of great significance in the field of anti-tumor. Summary of the Invention

[0006] The present invention discloses a class of aromatic hydrazide derivatives having a general structure of formula (I). Compounds of this structure show good application prospects in preventing and / or treating tumor diseases related to the MAPK pathway.

[0007] The present invention provides a compound represented by formula (I), its stereoisomers, tautomers or pharmaceutically acceptable salts thereof,

[0008] wherein Z is selected from oxygen or sulfur;

[0009] U1 is selected from nitrogen or CR 5d ; U2 is selected from nitrogen or CR 5a ; U3 is selected from nitrogen or CR 5b ; U4 is selected from nitrogen or CR 5c ;

[0010] Ring B is selected from substituted or unsubstituted aryl or heteroaryl;

[0011] R A Selected from -SO2NR a R b , -NR a SO2NR b R c , -NR a SO2R b , -C(O)NR a R b , -NR a C(O)NR b R c or -NR a C(O)R b ;

[0012] R 2a , R 2b , R 2c are each independently selected from hydrogen, deuterium, -OR a , -SR a , C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-10 membered heteroaryl, wherein alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl can be selected from deuterium, halogen, aldehyde, cyano, amino, hydroxy, oxo, C 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C6-10 substituted by one or more substituents in aryl or 5-10 membered heteroaryl;

[0013] R 3a , R 3b , R 5a , R 5b , R 5c , R 5d are independently selected from hydrogen, deuterium, halogen, aldehyde, cyano, amino, hydroxyl, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio, C 2-6 Alkenyl, C 2-6 Alkynyl or C 3-6 Cycloalkyl; or two adjacent substituents may together form a substituted or unsubstituted 3-10 membered ring structure;

[0014] X is selected from -O-, -CR a R b or -NR a ;

[0015] Ring A is selected from substituted or unsubstituted aryl or heteroaryl groups;

[0016] R a , R b , R c are each independently selected from hydrogen, deuterium, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Alkoxy, C 2-6 Alkenyl, C 2-6 Deuteroalkenyl, C 2-6 Alkynyl, C 2-6 Deuterated alkynyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-10 membered heteroaryl;

[0017] n is 1, 2, 3 or 4.

[0018] The present invention provides a compound represented by formula (II), its stereoisomers, tautomers or pharmaceutically acceptable salts thereof,

[0019] Wherein, U1 is selected from nitrogen or CR 5d ;

[0020] R A Selected from -SO2NR a R b , -NR a SO2NRb R c , -NR a SO2R b , -C(O)NR a R b , -NR a C(O)NR b R c or -NR a C(O)R b ;

[0021] R a , R b , R c are each independently selected from hydrogen, deuterium, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Alkoxy, C 2-6 Alkenyl, C 2-6 Deuteroalkenyl, C 2-6 Alkynyl, C 2-6 Deuterated alkynyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-10 membered heteroaryl;

[0022] R 2a , R 2b , R 2c are each independently selected from hydrogen, deuterium, -OR a , -SR a , C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-10 membered heteroaryl, wherein alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl can be selected from deuterium, halogen, aldehyde, cyano, amino, hydroxy, oxo, C 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 substituted by one or more substituents in aryl or 5-10 membered heteroaryl;

[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 are independently selected from hydrogen, deuterium, halogen, aldehyde, cyano, amino, hydroxyl, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio, C 2-6 Alkenyl, C 2-6 Alkynyl or C 3-6 or two adjacent substituents may together form a substituted or unsubstituted 3-10 membered ring structure.

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

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

[0026] R 2a , R 2b are each independently selected from hydrogen, deuterium, -OR a , -SR a , C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-10 membered heteroaryl, wherein alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl can be selected from deuterium, halogen, aldehyde, cyano, amino, hydroxy, oxo, C 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, C 5-10 substituted by one or more substituents in aryl or 5-10 membered heteroaryl;

[0027] R a Selected from hydrogen, deuterium, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 2-6 Alkenyl, C 2-6 Deuteroalkenyl, C 2-6 Alkynyl, C 2-6 Deuterated alkynyl or C 3-6 Cycloalkyl;

[0028] R3a , R 3b are each independently selected from hydrogen, deuterium, halogen or C 1-6 alkyl;

[0029] 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 independently selected from hydrogen, deuterium, halogen, aldehyde, cyano, amino, hydroxyl, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio, C 2-6 Alkenyl, C 2-6 Alkynyl or C 3-6 Cycloalkyl; or two adjacent substituents may together form a substituted or unsubstituted 3-10 membered ring structure;

[0030] R7 is selected from hydrogen, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl, C 2-6 Alkenyl or C 2-6 Alkynyl.

[0031] The present invention provides the following compounds, their stereoisomers, tautomers or pharmaceutically acceptable salts thereof,

[0032] The present invention provides a compound represented by formula (IV-I), its stereoisomers, tautomers or pharmaceutically acceptable salts thereof,

[0033] Wherein, U1 is selected from nitrogen or CR 5d ;

[0034] R 2b , R 3a , R 3b Selected from hydrogen, deuterium, halogen, C 1-6 Alkyl, C 1-6 Deuterated alkyl or C 1-6 alkyl halide;

[0035] 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 independently selected from hydrogen, deuterium, halogen, aldehyde, cyano, amino, hydroxyl, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio, C 2-6 Alkenyl, C 2-6 Alkynyl or C 3-6 Cycloalkyl; or two adjacent substituents may together form a substituted or unsubstituted 3-10 membered ring structure;

[0036] R7 is selected from hydrogen, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl, C 2-6 Alkenyl or C 2-6 Alkynyl;

[0037] R 8a , R 8b , R 8c , R 8d , R 8e are each independently selected from hydrogen, deuterium, halogen, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl or C 3-6 Cycloalkyl;

[0038] n is 1, 2, 3 or 4.

[0039] The present invention provides a compound represented by formula (IV-II), its stereoisomers, tautomers or pharmaceutically acceptable salts thereof,

[0040] wherein U1 is selected from nitrogen or CH;

[0041] R 2b Selected from hydrogen, deuterium or C 1-6 alkyl;

[0042] R 4a , R 4care independently selected from hydrogen, deuterium, halogen, aldehyde, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 Alkenyl, C 2-6 Alkynyl or C 3-6 Cycloalkyl;

[0043] R 5c , R 6b , R 6c are each independently selected from hydrogen, deuterium, halogen or C 1-6 alkyl;

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

[0045] R 8a , R 8b , R 8c , R 8d , R 8e are each independently selected from hydrogen, deuterium, halogen, C 1-6 Alkyl, C 1-6 Deuterated alkyl or C 1-6 alkyl halide;

[0046] n is 1, 2, 3 or 4.

[0047] The present invention provides a compound represented by formula (IV-III), its stereoisomers, tautomers or pharmaceutically acceptable salts thereof,

[0048] Among them, R 2b Selected from hydrogen or C 1-6 alkyl;

[0049] R 4c Selected from halogen, aldehyde, C 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 Alkenyl, C 2-6 Alkynyl or C 3-6 Cycloalkyl;

[0050] R7 is selected from C 1-6 Alkyl, C 1-6 Deuterated alkyl or C 3-6 Cycloalkyl;

[0051] R 8a , R 8b , R 8c , R 8d , R 8e are each independently selected from hydrogen, deuterium or C 1-6 alkyl;

[0052] n is 1, 2, 3 or 4.

[0053] The present invention provides the following compounds, their stereoisomers, tautomers or pharmaceutically acceptable salts thereof,

[0054] The present invention provides a compound represented by formula (V), its stereoisomers, tautomers or pharmaceutically acceptable salts thereof,

[0055] Wherein, U1 is selected from nitrogen or CR 5d ;

[0056] R 2b , R 3a , R 3b Selected from hydrogen, deuterium, halogen, C 1-6 Alkyl, C 1-6 Deuterated alkyl or C 1-6 alkyl halide;

[0057] 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 independently selected from hydrogen, deuterium, halogen, aldehyde, cyano, amino, hydroxyl, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio, C 2-6 Alkenyl, C 2-6 Alkynyl or C 3-6 Cycloalkyl; or two adjacent substituents may together form a substituted or unsubstituted 3-10 membered ring structure;

[0058] R7 is selected from hydrogen, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl, C 2-6 Alkenyl or C 2-6 Alkynyl;

[0059] R 8a , R8b , R 8c are each independently selected from hydrogen, deuterium, halogen, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl or C 3-6 Cycloalkyl;

[0060] n is 1, 2, 3 or 4.

[0061] The present invention provides the following compounds, their stereoisomers, tautomers or pharmaceutically acceptable salts thereof,

[0062] 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 or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0063] Use of any one of the compounds of the present invention, its stereoisomers, tautomers or pharmaceutically acceptable salts, or the pharmaceutical composition of the present invention in the preparation of drugs for preventing and / or treating tumors.

[0064] Use of any one of the compounds of the present invention, its stereoisomers, tautomers or pharmaceutically acceptable salts, or the pharmaceutical composition of the present invention, in the preparation of a medicament for preventing and / or treating diseases related to the MAPK pathway, particularly tumors.

[0065] The application of the present invention, wherein the tumor includes renal cancer, liver cancer, breast cancer, pancreatic cancer, prostate cancer, melanoma, leukemia, malignant lymphoma, ovarian cancer, head and neck cancer, lung cancer, colorectal cancer, bladder cancer, uterine cancer, etc.

[0066] Detailed Description of the Invention

[0067] All technical and scientific terms used in this specification have the same meanings as commonly understood by those skilled in the art.

[0068] The term "hydrogen" refers to H herein.

[0069] The term "deuterium" refers to D herein.

[0070] The term "nitrogen" refers to N herein.

[0071] The term "oxygen" refers to O herein.

[0072] The term "sulfur" refers to S herein.

[0073] The term "cyano" refers herein to -CN.

[0074] The term "hydroxy" refers herein to -OH.

[0075] The term "halogen" as used herein refers to -F, -Cl, -Br and -I.

[0076] The term "aldehyde group" refers herein to -CHO.

[0077] The term "amino" or "amine" refers interchangeably herein 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, aryl, heteroaryl, 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 halogen, cyano, alkenyl, alkynyl, alkyl, alkoxy, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, urea, carbonyl, carboxylate, amine, and amide.

[0078] The term "alkyl" herein refers to a saturated aliphatic hydrocarbon group having 1 to 15 carbon atoms, preferably 1 to 10 carbon atoms, more preferably 1 to 6 carbon atoms, and includes straight-chain and branched hydrocarbon groups. When the number of carbon atoms before the alkyl group is specified, such as C 1-6 Alkyl refers to an alkyl group having 1 to 6 carbon atoms. Non-limiting examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, neopentyl, n-hexyl, etc. Alkyl groups as 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, acyloxy, oxo, amide, ester, amine, sulfonyl, sulfinyl, cycloalkyl, heterocyclyl, cycloalkenyl, heterocycloalkyl, alkenyl, alkenyloxy, alkynyl, cycloalkyloxy, heterocycloalkyloxy, aryloxy, heteroaryloxy, aryl or heteroaryl.

[0079] The term "deuterated alkyl" herein refers to an alkyl group obtained by substituting the aforementioned "alkyl" group with deuterium. Non-limiting examples of deuterated alkyl groups include deuterated methyl, deuterated ethyl, and the like.

[0080] The term "haloalkyl" herein refers to an alkyl group obtained by substituting the aforementioned "alkyl" with a halogen, wherein the halogen includes fluorine, chlorine, bromine, iodine and the like.

[0081] The term "alkoxy" refers to a group of the formula -OR a Group, where R a When the number of carbon atoms before the alkoxy group is limited, such as C 1-6Alkoxy refers to an alkoxy group containing 1 to 6 carbon atoms. Examples of alkoxy include, but are not limited to, methoxy, ethoxy, isopropoxy, n-propoxy, n-butoxy, tert-butoxy, n-pentoxy, and the like.

[0082] Alkenyl is an unsaturated hydrocarbon group containing a carbon-carbon double bond. The term "alkenyl" herein refers to an alkyl group containing a carbon-carbon double bond in the molecule, wherein the alkyl group is as defined above. Alkenyl herein has 2 to 10 carbon atoms, preferably 2 to 8 carbon atoms, more preferably 2 to 6 carbon atoms. When the number of carbon atoms in the alkenyl group is specified, such as C 2-6 Alkenyl refers to that the alkenyl contains 2-6 carbon atoms.Alkenyl described herein can be optionally substituted by one or more of the following substituents: deuterium, fluorine, chlorine, bromine, iodine, cyano group, nitro, hydroxyl, carboxyl, amino, oxo, alkyl, alkoxy, acyl, amide, ester group, amido, sulfonyl, sulfinyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, cycloalkyloxy, sulfydryl, alkyl mercapto, deuterated alkyl mercapto, sulfone, sulfoxide, silicon, phosphono, deuterated alkyl, heterocyclic radical, aryl, heteroaryl, alkynyl, alkenyl, arylalkyl. Non-limiting examples of alkenyl include vinyl, propenyl, allyl, isopropenyl, butenyl, isobutenyl etc.

[0083] Alkynyl is an unsaturated hydrocarbon group containing a carbon-carbon triple bond. The term "alkynyl" herein refers to an alkyl group containing a carbon-carbon triple bond in the molecule, wherein the alkyl group is as defined above. Alkynyl herein has 2 to 10 carbon atoms, preferably 2 to 8 carbon atoms, more preferably 2 to 6 carbon atoms. When the number of carbon atoms in front of the alkynyl group is limited, such as C 2-6 Alkynyl refers to that the alkynyl contains 2-6 carbon atoms.Alkynyl can be optionally substituted by one or more following substituents: deuterium, fluorine, chlorine, bromine, iodine, cyano group, nitro, hydroxyl, carboxyl, amino, alkyl, oxo, alkoxyl, acyl group, amide groups, ester groups, amido, sulfonyl, sulfinyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, cycloalkyloxy, sulfydryl, alkyl mercapto, deuterated alkyl mercapto, sulfone group, sulfoxide group, silicon radical, phosphono, deuterated alkyl, heterocyclic radical, aryl, heteroaryl, alkynyl, alkenyl, arylalkyl.Alkynyl non-limiting examples comprises ethynyl, 1-propynyl, 2-propynyl, 1-, 2-or 3-butynyl etc.

[0084] The term "cycloalkyl" refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon group consisting only of carbon atoms and hydrogen atoms, which may include a fused ring system, a spiro ring system or a bridged ring system, having 3 to 15 carbon atoms, preferably 3 to 10 carbon atoms, more preferably 3 to 8 carbon atoms, most preferably 3 to 6 carbon atoms, and which is saturated or unsaturated and can be connected to the rest of the molecule by a single bond via any suitable carbon atom. When the cycloalkyl group is preceded by a carbon atom number limit, such as C 3-6Cycloalkyl means that the cycloalkyl group contains 3-6 carbon atoms. Non-limiting examples of cycloalkyl include, but are 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, etc. The cycloalkyl groups described herein may be optionally substituted with one or more of the following substituents: deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, hydroxy, oxo, carboxyl, amino, alkyl, alkoxy, acyl, amide, ester, amine, sulfonyl, sulfinyl, cycloalkyl, heterocyclyl, cycloalkenyl, heterocycloalkyl, alkenyl, alkynyl, cycloalkoxy, aryl, and heteroaryl.

[0085] 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. The aromatic or non-aromatic ring may be a 3- to 10-membered monocyclic ring, a 4- to 20-membered spirocyclic ring, a fused ring, or a bridged ring, preferably a 3- to 12-membered heterocyclic ring. The optionally substituted nitrogen or sulfur atoms in the heterocyclyl ring may be oxidized to various oxidation states. Non-limiting examples of heterocyclyl groups 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. The heterocyclyl groups described herein may be optionally substituted with one or more of the following substituents: deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, hydroxy, carboxyl, amino, alkyl, alkoxy, acyl, amide, ester, amine, sulfonyl, sulfinyl, cycloalkyl, heterocyclyl, cycloalkenyl, heterocycloalkyl, alkenyl, alkynyl, cycloalkoxy, aryl, and heteroaryl.

[0086] The term "aryl" herein refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., rings sharing adjacent pairs of carbon atoms) group having a conjugated π electron system, preferably 6-10 members, such as phenyl and naphthyl, more preferably phenyl. The aryl ring may be fused to a heteroaryl, heterocyclyl, or cycloalkyl ring, wherein the ring connected to the parent core structure is the aryl ring. The aryl groups described herein may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups: deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, hydroxyl, carboxyl, amino, alkyl, alkoxy, acyl, amide, ester, sulfonyl, sulfinyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, alkenyl, alkynyl, heterocyclyl, cycloalkoxy, aryl, and heteroaryl.

[0087] 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, triazole, 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, oxo, alkyl, alkoxy, acyl, acyloxy, amide, ester, amine, sulfonyl, sulfinyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, alkenyl, alkynyl, heterocyclyl, cycloalkoxy, aryl, and heteroaryl.

[0088] 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 14Compounds 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.

[0089] 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.

[0090] The term "compound of the present invention" (unless otherwise specifically stated) refers to the compounds of formula (I) to formula (V) 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.

[0091] 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.

[0092] The term "optionally substituted by..." means that the structure is unsubstituted or substituted by one or more substituents described herein. 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 on the same part) is chemically allowed, 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 substituent and can therefore be the same or different.

[0093] 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.

[0094] 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.

[0095] The term "tautomer" as used herein refers to structural isomers having 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 -NH- and -N- portions of the ring, such as pyrazoles, imidazoles, benzimidazoles, triazoles, and tetrazoles. Valence tautomers include interconversion due to reorganization of some of the bonding electrons.

[0096] 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.

[0097] 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, kidney cancer, gallbladder cancer, pancreatic cancer, liver cancer, prostate cancer, uterine cancer, testicular cancer, ovarian cancer, renal cell carcinoma, colorectal cancer, bladder cancer, renal pelvis-ureter cancer, melanoma, skin cancer and other solid tumors. DETAILED DESCRIPTION

[0098] 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.

[0099] 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 can usually be obtained commercially, for example, by It can be purchased from companies such as TCI, or prepared by methods well known to those skilled in the art.

[0100] The compound of formula (III) of the present invention, its stereoisomers, tautomers or pharmaceutically acceptable salts can be prepared by the following process:

[0101] The compound of formula (III) of the present invention can be prepared according to the above process: under alkaline conditions, a fluorobenzene derivative A and an aniline derivative B are reacted to produce a diphenylamine derivative C; the diphenylamine derivative C is subjected to esterification and oxidation reactions to produce an aldehyde derivative D; the aldehyde derivative D is reacted with an arylsulfonylhydrazine to produce a hydrazine derivative E; under alkaline conditions, the hydrazine derivative E is reacted with an arylboronic acid derivative F to produce a compound G; under acidic conditions, the compound G is deprotected to produce an ester derivative H; under alkaline conditions, the ester derivative H is reacted with a corresponding sulfonyl compound to produce a compound I; compound I is hydrolyzed in the presence of a hydroxide to produce a benzoic acid derivative J; in the presence of a condensing agent, the benzoic acid derivative J is reacted with a corresponding hydrazine or hydrazine hydrochloride to produce a compound of general formula III. The substituents are as defined above.

[0102] 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.

[0103] The structures of the following example compounds were characterized by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). The compounds were dissolved in appropriate deuterated reagents and analyzed at ambient temperature using TMS as an internal standard using a Bruker Ascend 400 MHz NMR spectrometer. 1H-NMR analysis. NMR chemical shifts (δ) are in ppm and are referred to using the following abbreviations: s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; brs, broad singlet.

[0104] 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.).

[0105] Preparation of intermediates

[0106] Preparation of Intermediate 1

[0107] Step 1: Preparation of 2,3,4-trifluoro-5-iodobenzoic acid

[0108] To a 3L three-necked flask, add acetic acid (928mL) and acetic anhydride (538.0g) and cool to 0°C in an ice bath. Slowly add concentrated sulfuric acid (2116.0g) dropwise to the system, maintaining the system temperature above 40°C. After completion of the addition, cool the system to room temperature again. Then add 2,3,4-trifluorobenzoic acid (115.0g), iodine (66.9g), and manganese dioxide (68.7g). After addition, raise the temperature to 50°C and react for 3 hours. Stop heating, allow the system to cool to room temperature, then add additional iodine (66.9g) and manganese dioxide (68.7g), then raise the temperature to 50°C and react for 3 hours. Stop heating, allow the system to cool to room temperature, then add additional iodine (33.4g) and manganese dioxide (34.4g), then raise the temperature to 50°C again and react for 21 hours. Stop the reaction, allow the system to cool to room temperature, and then slowly pour into ice water to quench the reaction. Extract with dichloromethane, combine the organic phases, wash with aqueous sodium sulfite and saturated brine, and dry over anhydrous sodium sulfate. After concentrating the organic phase, add n-heptane and stir in an ice bath to precipitate a large amount of solid. Filter with suction and wash the filter cake with n-heptane to obtain 178.1 g of the product as a white solid. ESI-MS: [MH] - =300.9.

[0109] Step 2: Preparation of 2,3,4-trifluoro-5-vinylbenzoic acid

[0110] In a 3L three-necked flask, 2,3,4-trifluoro-5-iodobenzoic acid (89.0 g), potassium vinyl trifluoroborate (47.4 g), potassium phosphate (93.8 g), methanol (1780 mL), water (890 mL) and [1,1′-bis(diphenylphosphino)ferrocene] palladium dichloride (21.6 g) were added in sequence. The system was replaced with a nitrogen atmosphere and heated to 60°C for 3 hours. After the system was cooled to room temperature, it was filtered through celite, the filtrate was collected, concentrated, and 1M HCl was added to adjust the pH of the system to 4-5. Then, it was extracted with tert-methyl ether, the organic phases were combined, washed with saturated brine, dried and concentrated. n-heptane and dichloromethane were added to the concentrate, stirred at room temperature for 2 hours, filtered, and the filter cake was washed with n-heptane to obtain 48.2 g of a white solid. ESI-MS: [MH] - =201.0.

[0111] Step 3: Preparation of 3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-5-vinylbenzoic acid

[0112] In a 3L three-necked flask, 2,3,4-trifluoro-5-vinylbenzoic acid (65.0g), 2-fluoro-4-iodoaniline (76.3g) and tetrahydrofuran (1300mL) were added, and then the system was replaced with a nitrogen environment and cooled to -15 to -20°C. Lithium bis(trimethylsilyl)amide (1M, 965mL) 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 1M hydrochloric acid was added to adjust the pH of the system to 1 to 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, precipitate the solid, filter, and obtain 99.0g of off-white solid. ESI-MS: [MH] - =418.0.

[0113] Step 4: Preparation of methyl 3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-5-vinylbenzoate

[0114] 3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-5-vinylbenzoic acid (20.0 g) and N,N-dimethylformamide (200 mL) were added to a 1L three-necked flask and stirred at room temperature until the solution became clear. Potassium carbonate (7.9 g) and iodomethane (8.1 g) were added to the system, and the system was then reacted at room temperature for 12 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 1 h. Filtered and the filter cake was washed with 250 mL of ethanol to obtain 15.1 g of a light yellow solid.

[0115] Step 5: Preparation of methyl 3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-5-formylbenzoate

[0116] In a 1L three-necked flask, methyl 3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-5-vinylbenzoate (11.6g) and tetrahydrofuran (200mL) were added and stirred in an ice bath until dissolved. Potassium osmate dihydrate (494mg) was added to the system and the reaction was continued by stirring in an ice bath. After 15min, water (200mL), sodium periodate (17.2g) and 2,6-lutidine (2.9g) 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 with saturated brine, and the organic phases were concentrated to obtain a solid crude product. The crude product was washed with anhydrous ethanol and filtered to obtain 9.0g of a light brown 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(d d, J1=10.4Hz, J2=1.6Hz,1H),7.54-7.51(m,1H),7.13-7.07(m,1H),3.86(s,3H).ESI-MS:[M+H] + =436.0.

[0117] Step 6: Preparation of methyl (E)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-5-((2-toluenesulfonylhydrazono)methyl)benzoate

[0118] 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 h. The reaction was then cooled to 0°C in an ice bath, resulting in the precipitation of a large amount of solid. The solid was filtered, and the filter cake was washed with ethanol to obtain 53.1 g of a light yellow solid. 1 H 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),6.92-6.89(m,1H),3.85(s,3H),2.37(s,3H).ESI-MS:[M+H] + =604.0.

[0119] Step 7: Preparation of N-(2,4-dimethoxybenzyl)-3-fluoro-4-iodopyridin-2-amine

[0120] 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 2 hours. After the system cools to room temperature, it is poured into water and extracted with tertiary methyl ether. The organic phases are combined, washed with saturated brine, dried, and concentrated to obtain a crude product. The concentrate is slurried with tertiary 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=5.6Hz,1H),7.11-7.08(m,1H),7.03(d,J=8.4Hz,1H),6.89-6.86(m,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).

[0121] Step 8: Preparation of N-(2,4-dimethoxybenzyl)-3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine

[0122] 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)ferrocene]palladium 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 mixture is poured into water and extracted with ethyl acetate. The organic phases are combined, dried over anhydrous sodium sulfate, and concentrated. The concentrate is then diluted with n-hexane, dried, filtered through celite, and the filtrate is concentrated to obtain the product. The product from this step is carried on directly to the next step without further purification. 1H NMR (400MHz, DMSO-d6): δ7.75 (dd, J1=4.8Hz, J2=0.8Hz, 1H), 7.01 (d, J=8.4Hz, 1H), 6.87-6.84 (m, 1H), 6.59-6.56 (m, 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).

[0123] Step 9: Preparation of (2-((2,4-dimethoxybenzyl)amino)-3-fluoropyridin-4-yl)boronic acid

[0124] In a 2L single-necked bottle, N-(2,4-dimethoxybenzyl)-3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine and THF (500mL) were added and stirred until clear. 1M aqueous hydrochloric acid solution was slowly added to the system and the reaction was stirred at room temperature for 1h. The system was extracted with petroleum ether and the aqueous phase was collected. Methylboric acid (17.4g) was added to the aqueous phase and the reaction was stirred at room temperature for 1h. The system was then extracted with a mixed solution of ethyl acetate / petroleum ether (3 / 1) and the aqueous phase was collected. Ethyl acetate was added to the aqueous phase and the pH was adjusted to 8 with aqueous sodium bicarbonate solution. The organic phase was collected, washed with water, dried and concentrated to give 37.0g of an off-white solid. ESI-MS: [M+H] + =307.1.

[0125] Step 10: Preparation of methyl 5-((2-((2,4-dimethoxybenzyl)amino)-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoate

[0126] In a 1L single-necked flask, (2-((2,4-dimethoxybenzyl)amino)-3-fluoropyridin-4-yl)boric acid (15.0 g), (E)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-5-((2-toluenesulfonylhydrazono)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 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 then purified by silica gel column chromatography to obtain 17.9 g of a light yellow liquid. 1H NMR (400MHz, DMSO-d6): 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=3.0Hz,1H),7.39(dd,J1=8.4Hz,J2=0.8Hz,1H),7.03(d,J=8.4Hz,1H),6.92-6.88(m ,1H),6.80-6.75(m,1H),6.54(d,J=2.4Hz,1H),6.42(dd,J1=8.4Hz,J2=2.4Hz,1H),6.38(t,J=4 .8Hz,1H),4.44(d,J=6.0Hz,2H),4.01(s,2H),3.80(d,J=1.2Hz,6H),3.72(s,3H).ESI-MS:[M+H] + =682.1.

[0127] Step 11: Preparation of methyl 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoate

[0128] In a 500mL single-necked bottle, 5-((2-((2,4-dimethoxybenzyl)amino)-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoic acid methyl ester (10.0g) and dichloromethane (150mL) were added and stirred at room temperature until clear. The system was then placed in an ice bath and trifluoroacetic acid (150mL) 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 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 5.8g 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=1.6Hz,1H),7.39 (dd,J1=8.4Hz, J2=1.2Hz,1H),6.80-6.74(m,1H),6.40(t,J=4.8Hz,1H),6.22(s,2H),3.99(s,2H),3.79(s,3H).ESI-MS:[M+H] + =532.0.

[0129] Preparation of Intermediate 2

[0130] Step 1: Preparation of tert-butyl hydrazinecarboxylate

[0131] Add hydrazine hydrate (11.3 g) and isopropanol (100 mL) to a 250 mL single-necked flask. Replace the atmosphere with nitrogen, cool to 0°C, and dropwise add a solution of di-tert-butyl dicarbonate (17.4 g) in isopropanol (20 mL). After addition, warm the system to room temperature and stir for 2 h. Concentrate the reaction mixture under reduced pressure to yield 10.3 g of a colorless oil.

[0132] Step 2: Preparation of tert-butyl 2-allylhydrazine-1-carboxylate

[0133] To a 250 mL single-necked flask, add tert-butyl hydrazinecarboxylate (10.3 g), dimethyl sulfoxide (50 mL), triethylamine (6.5 g), and allyl bromide (7.8 g). Replace the atmosphere with nitrogen and stir at room temperature for 3 h. Add water (100 mL) and extract with ethyl acetate. The organic phases are combined, dried, and concentrated. The concentrate is purified by silica gel column chromatography to yield 3.2 g of a light yellow oil.

[0134] Step 3: Preparation of allylhydrazine hydrochloride

[0135] To a 100 mL single-necked flask, add tert-butyl 2-allylhydrazine-1-carboxylate (3.2 g) and tetrahydrofuran (6 mL). Slowly add concentrated hydrochloric acid (6 mL) dropwise with stirring at room temperature. Continue stirring at room temperature for 3 h. The reaction solution is concentrated under reduced pressure to yield 1.9 g of an off-white solid.

[0136] Preparation of Intermediate 3

[0137] Preparation of N-ethyl-2-oxoxazoline-3-sulfonamide

[0138] 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).

[0139] Preparation of Intermediate 4

[0140] Preparation of N-cyclopropyl-2-oxoxazoline-3-sulfonamide

[0141] Under ice bath conditions, 500mL of acetonitrile and chlorosulfonyl isocyanate (11.6g) were added to a 1L single-necked flask. 2-bromoethanol (10.3g) 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 (50.0g) was then slowly added to the system and stirred for 10min, after which cyclopropylamine (4.7g) 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 mixture was filtered and the filtrate was concentrated to obtain a light orange solid. Ethyl acetate was added to dissolve most of the solid, washed with saturated brine, the organic phase was separated and dried over anhydrous sodium sulfate, filtered and the solvent was removed by spinning to obtain a crude product. The crude product was slurried with tert-methyl ether to obtain 4.4g 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).

[0142] Preparation of Intermediate 5

[0143] Preparation of N-methyl-d3-2-oxoxazoline-3-sulfonamide

[0144] 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 15h. Filtered and the filtrate was concentrated. Water was added to dilute the concentrate, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. Filtered and the solvent was removed by spinning to obtain 11.9g of a light yellow solid. 1 H NMR (400MHz, DMSO-d6, δ): 8.17 (s, 1H), 4.42-4.38 (m, 2H), 3.96-3.93 (m, 2H).

[0145] Compound preparation

[0146] Example 1

[0147] Preparation of N'-n-propyl-3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzohydrazide

[0148] To a 100 mL single-necked bottle, 3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzoic acid (500 mg), N,N-dimethylformamide (10 mL), 1-hydroxybenzotriazole (170 mg), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (470 mg) were added and stirred at room temperature for 1 hour. n-Propylhydrazine hydrochloride (170 mg) and N,N-diisopropylethylamine (320 mg) were added to the system, and the reaction was continued by stirring at room temperature for 1.5 hours. The reaction solution was slowly added to a saturated sodium bicarbonate solution, extracted with ethyl acetate, and the organic phases were combined, dried, and concentrated. The concentrate was purified by silica gel column chromatography to obtain 190 mg of an off-white solid. ESI-MS: [M+H] + =667.1.

[0149] Example 2

[0150] Preparation of N'-isopropyl-3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzohydrazide

[0151] To a 100 mL single-necked flask, 3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzoic acid (400 mg), N,N-dimethylformamide (20 mL), 1-hydroxybenzotriazole (133 mg), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (188 mg) were added and stirred at room temperature for 1 hour. Isopropylhydrazine hydrochloride (145 mg) and N,N-diisopropylethylamine (338 mg) were added to the system, and the reaction was continued by stirring at room temperature for 2 hours. The reaction solution was slowly added to a saturated sodium bicarbonate solution, extracted with ethyl acetate, and the organic phases were combined, dried, and concentrated. The concentrate was purified by silica gel column chromatography to obtain 130 mg of an off-white solid. 1 H NMR (400MHz, DMSO-d6): δ10.42(s,1H),10.12(d,J=6.4Hz,1H),8.83(s,1H),8.02(d, J=5.2Hz,1H),7.58(dd,J=10.8,2.0Hz,1H),7.48(d,J=7.2Hz,1H),7.35(d,J=8.4Hz,1 H),7.07-7.00(m,1H),6.96(t,J=4.8Hz,1H),6.69-6.63(m,1H),4.99-4.97(m,1H),4. 08(s,2H),3.01-2.94(m,1H),2.52-2.51(m,3H),0.96(d,J=6.0Hz,6H).ESI-MS:[M+H] + =667.1.

[0152] Example 3

[0153] Preparation of N'-tert-butyl-3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzohydrazide

[0154] To a 100 mL single-necked flask, 3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzoic acid (500 mg), N,N-dimethylformamide (15 mL), 1-hydroxybenzotriazole (170 mg), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (470 mg) were added and stirred at room temperature for 1 hour. Tert-butylhydrazine hydrochloride (200 mg) and N,N-diisopropylethylamine (320 mg) were added to the system, and the reaction was stirred at room temperature for another 2 hours. The reaction solution was slowly added to a saturated sodium bicarbonate solution, extracted with ethyl acetate, and the organic phases were combined, dried, and concentrated. The concentrate was purified by silica gel column chromatography to obtain 180 mg of an off-white solid. 1 H NMR (400MHz, DMSO-d6): δ10.43(s,1H),9.99(d,J=6.8Hz,1H),8.67(s,1H),8.0 2(d,J=4.8Hz,1H),7.58(dd,J=10.8,2.0Hz,1H),7.51(d,J=6.8Hz,1H),7.34(d ,J=8.4Hz,1H),7.06-7.02(m,1H),6.96(t,J=4.4Hz,1H),6.67-6.60(m,1H),4. 92(d,J=7.2Hz,1H),4.10(s,2H),2.52-2.51(m,3H),1.00(s,9H).ESI-MS:[M+H] + =681.1.

[0155] Example 4

[0156] Preparation of N'-hydroxyethyl-3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzohydrazide

[0157] In a 100 mL single-necked bottle, 3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzoic acid (1.0 g), N,N-dimethylformamide (20 mL), 1-hydroxybenzotriazole (266 mg), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (629 mg) were added and stirred at room temperature for 1 hour. Hydroxyethylhydrazine hydrochloride (374 mg) and N,N-diisopropylethylamine (423 mg) were added to the system and the reaction was continued to stir at room temperature for 2 hours. The reaction solution was slowly added to a saturated sodium bicarbonate solution, extracted with ethyl acetate, and the organic phases were combined, dried, and concentrated. The concentrate was purified by silica gel column chromatography to obtain 350 mg of an off-white solid. ESI-MS: [M+H] + =669.0.

[0158] Example 5

[0159] Preparation of 3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzohydrazide

[0160] In a 100 mL single-necked bottle, 3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzoic acid (400 mg), N,N-dimethylformamide (20 mL), 1-hydroxybenzotriazole (133 mg) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (188 mg) were added and stirred at room temperature for 1 hour. Hydrazine dihydrochloride (276 mg) and N,N-diisopropylethylamine (680 mg) were added to the system and the reaction was continued to stir at room temperature for 2 hours. The reaction solution was slowly added to a saturated sodium bicarbonate solution, extracted with ethyl acetate, and the organic phases were combined, dried, and concentrated. The concentrate was purified by silica gel column chromatography to obtain 100 mg of a light yellow solid. ESI-MS: [M+H] + =625.0.

[0161] Example 6

[0162] Preparation of N'-cyclopropyl-3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzohydrazide

[0163] To a 100 mL single-necked bottle, 3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzoic acid (700 mg), N,N-dimethylformamide (15 mL), 1-hydroxybenzotriazole (187 mg), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (441 mg) were added and stirred at room temperature for 1 hour. Cyclopropylhydrazine hydrochloride (375 mg) and N,N-diisopropylethylamine (445 mg) were added to the system and the reaction was continued to stir at room temperature for 2 hours. The reaction solution was slowly added to a saturated sodium bicarbonate solution, extracted with ethyl acetate, and the organic phases were combined, dried, and concentrated. The concentrate was purified by silica gel column chromatography to obtain 350 mg of an off-white solid. 1 H NMR (400MHz, DMSO): δ10.43 (s, 1H), 10.20 (d, J = 6.0Hz, 1H), 8.95 (s, 1H), 8.02 (d,J=4.4Hz,1H),7.59(dd,J=10.8,2.0Hz,1H),7.49(d,J=6.8Hz,1H),7.36(d, J=8.4Hz,1H),7.04(d,J=4.4Hz,1H),6.95(brs,1H),6.71-6.66(m,1H),5.67- 5.64(m,1H),4.07(s,2H),2.52-2.50(m,4H),0.39-0.38(m,4H).ESI-MS:[M+H] + =665.1.

[0164] Example 7

[0165] Step 1: Preparation of methyl 3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzoate

[0166] To a 100 mL single-necked flask, add methyl 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoate (460 mg), N,N-dimethylformamide (10 mL), and pyridine (680 mg), and cool to -20°C. Slowly add methylaminosulfonyl chloride (560 mg) dropwise to the system, and allow to react for 30 minutes. The reaction solution is poured into water and stirred at room temperature for 10 minutes. Filter with suction, dissolve the filter cake in ethyl acetate, wash the organic phase with saturated brine, dry, and concentrate to obtain 531 mg of an off-white solid. ESI-MS: [M+H] + =625.0.

[0167] Step 2: Preparation of 3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzoic acid

[0168] Add the product from the previous step (400 mg) and tetrahydrofuran (6 mL) to a 100 mL single-necked flask and stir at room temperature until dissolved. Add a 3 mL solution of lithium hydroxide monohydrate (81 mg) dropwise to the system. Continue stirring at room temperature for 3 h. Concentrate under reduced pressure to remove the tetrahydrofuran, add water to the system, adjust the pH to 2-3 with 1 M dilute hydrochloric acid, and stir for 10 min. Filter, collect the filter cake, wash with water, and dry to obtain 330 mg of an off-white solid. ESI-MS: [M+H] + =611.0.

[0169] Step 3: Preparation of N'-allyl-3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzohydrazide

[0170] To a 100 mL single-necked flask, add the product from the previous step (700 mg) and N,N-dimethylformamide (20 mL) and cool to 0°C in an ice bath. Then add 1-hydroxybenzotriazole (230 mg) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (660 mg), stir and react for 5 minutes, then warm to room temperature and react for 2 hours. Add allylhydrazine hydrochloride (250 mg) and N,N-diisopropylethylamine (590 mg) to the system and continue stirring at room temperature for 2 hours. The reaction solution is slowly added to a saturated sodium bicarbonate solution, extracted with ethyl acetate, and the organic phases are combined, dried, and concentrated. The concentrate is purified by preparative HPLC to yield 200 mg of a white solid. 1 H NMR (400MHz, DMSO-d6): δ10.42(s,1H),10.11(d,J=6.3Hz,1H),8.81(s,1H),8.02(d,J=4.9H z,1H),7.58(dd,J=10.8,1.8Hz,1H),7.40(d,J=6.9Hz,1H),7.35(d,J=8.5Hz,1H),7.04-6.9 5(m,2H),6.68-6.62(m,1H),5.85-5.75(m,1H),5.32-5.28(m,1H),5.10(d,J=17.3Hz,1H),5 .02(d,J=10.2Hz,1H),4.06(s,2H),3.34(t,J=5.5Hz,2H),2.51-2.50(m,3H).ESI-MS:[M+H] + =665.1.

[0171] Example 8

[0172] Step 1: Preparation of tert-butyl 2-allyl-2-methylhydrazine-1-carboxylate

[0173] To a 50 mL single-necked flask, add tert-butyl 2-allylhydrazine-1-carboxylate (800 mg), potassium carbonate (1.3 g), iodomethane (979 mg), and N,N-dimethylformamide (5 mL). Heat to 30°C and stir for 4.5 hours. Pour the reaction solution into water and extract with ethyl acetate. The organic phases are combined, dried, and concentrated to yield 800 mg of a light yellow-green liquid. ESI-MS: [M+H] + =187.1.

[0174] Step 2: Preparation of 1-allyl-1-methylhydrazine hydrochloride

[0175] The product from the previous step (800 mg) and a 4M solution of hydrogen chloride in ethyl acetate (5 mL) were added to a 50 mL single-necked bottle and stirred at room temperature for 2 h. The reaction solution was concentrated to obtain 500 mg of a yellow viscous product. ESI-MS: [M+H] + =87.1.

[0176] Step 3: Preparation of N'-methyl-N'-allyl-3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzohydrazide

[0177] To a 100 mL single-necked flask, add 3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzoic acid (400 mg) and N,N-dimethylformamide (8 mL) and cool to 0°C in an ice bath. Then, add 1-hydroxybenzotriazole (130 mg) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (380 mg). After stirring for 5 minutes, warm the mixture to room temperature and allow to react for 2 hours. Then, add 1-allyl-1-methylhydrazine hydrochloride (500 mg) and N,N-diisopropylethylamine (250 mg). After addition, continue stirring at room temperature for 2 hours. The reaction mixture is slowly added to a saturated sodium bicarbonate solution, extracted with ethyl acetate, and the organic phases are combined, dried, and concentrated. The concentrate is purified by silica gel column chromatography to yield 88 mg of an off-white solid. 1H NMR (400MHz, DMSO-d6): δ10.43(s,1H),9.48(s,1H),8.54(s,1H),8.02(d,J=4.0Hz ,1H),7.58(dd,J=10.8,2.0Hz,1H),7.35-7.33(m,2H),7.04(brs,1H),6.93(brs,1H ),6.65-6.60(m,1H),5.81-5.71(m,1H),5.10(dd,J=17.2,1.6Hz,1H),4.98(dd,J= 10.4,1.6Hz,1H),4.07(s,2H),3.34-3.33(m,2H),2.51-2.50(m,6H).ESI-MS:[M+H] + =679.1.

[0178] Example 9

[0179] Preparation of N'-allyl-3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-ethynylphenyl)amino)benzohydrazide

[0180] To a 100 mL single-necked flask, N'-allyl-3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzohydrazide (150 mg), cuprous iodide (1 mg), triethylamine (0.5 mL), tetrahydrofuran (5 mL), trimethylsilylacetylene (100 mg), and bistriphenylphosphine palladium dichloride (8 mg) were added. The atmosphere was replaced with nitrogen and the reaction was stirred at room temperature for 1 h. Subsequently, a solution of tetrabutylammonium fluoride in tetrahydrofuran (1 M, 3 mL) was added to the system and the reaction was stirred at room temperature for another 1 h. The mixture was filtered, the filter cake was washed with ethyl acetate, the organic phases were combined, washed with water, dried, and concentrated. The concentrate was purified by silica gel column chromatography to obtain the product. ESI-MS: [M+H] + =563.2.

[0181] Example 10

[0182] Preparation of N'-allyl-3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-vinylphenyl)amino)benzohydrazide

[0183] In a 50 mL single-necked bottle, add N'-allyl-3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzohydrazide (150 mg), potassium vinyl trifluoroborate (61 mg), potassium carbonate (94 mg), 1,4-dioxane (5 mL), water (1.5 mL), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (8 mg). The atmosphere is replaced with nitrogen, and the temperature is raised to 95°C and stirred for 1 hour. After cooling the reaction solution to room temperature, it is poured into water to quench the reaction. The aqueous phase is extracted with ethyl acetate, and the organic phases are combined, dried, and concentrated. The concentrate is purified by silica gel column chromatography to obtain the product. ESI-MS: [M+H] + =565.2.

[0184] Example 11

[0185] Step 1: Preparation of methyl 5-((2-((N-cyclopropylaminosulfonyl)amino)-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoate

[0186] To a 100 mL single-necked flask, add N-cyclopropyl-2-oxoxazoline-3-sulfonamide (970 mg), methyl 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoate (500 mg), triethylamine (570 mg), and 1,2-dichloroethane (25 mL). Heat to 80°C and react for 4 hours. After cooling to room temperature, the reaction solution is poured into a water / tetrahydrofuran mixture (90 / 10, v / v) and extracted with ethyl acetate. The organic phases are combined, dried, and concentrated to obtain 600 mg of a brown solid. ESI-MS: [M+H] + =651.0.

[0187] Step 2: Preparation of 5-((2-((N-cyclopropylaminosulfonyl)amino)-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoic acid

[0188] In a 50 mL single-necked flask, add the product from the previous step (600 mg), tetrahydrofuran (30 mL), and water (12 mL) in sequence and stir until dissolved. Add lithium hydroxide monohydrate (193 mg) to the system and stir at room temperature for 1.5 hours. Add aqueous hydrochloric acid (1 M) to adjust the pH to 4-5 and concentrate under reduced pressure to remove tetrahydrofuran. Add water to the concentrate, filter, and collect the filter cake to obtain 580 mg of a brown solid. ESI-MS: [M+H] + =637.0.

[0189] Step 3: Preparation of N'-allyl-3,4-difluoro-5-((3-fluoro-2-((N-cyclopropylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzohydrazide

[0190] The product from the previous step (580 mg), N,N-dimethylformamide (20 mL), 1-hydroxybenzotriazole (216 mg), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (305 mg) were added to a 100 mL single-necked flask and stirred at room temperature for 1 hour. Allylhydrazine hydrochloride (265 mg) and N,N-diisopropylethylamine (793 mg) were added to the system and the reaction was continued to stir at room temperature for 30 minutes. The reaction solution was slowly added to a saturated sodium bicarbonate aqueous solution and extracted with ethyl acetate. The organic phases were combined, dried, and concentrated. The concentrate was purified by silica gel column chromatography to obtain the product. ESI-MS: [M+H] + =691.1.

[0191] Example 12

[0192] Step 1: Preparation of methyl 5-((2-((N-ethylaminosulfonyl)amino)-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoate

[0193] N-ethyl-2-oxoxazoline-3-sulfonamide (5.5 g), methyl 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoate (1.5 g), triethylamine (3.1 g), and 1,2-dichloroethane (50 mL) were added to a 100 mL single-necked flask and heated to 80°C for 4.5 h. After the system cooled to room temperature, it 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, and concentrated to obtain 1.0 g of a yellow oil. ESI-MS: [M+H] + =639.0.

[0194] Step 2: Preparation of 5-((2-((N-ethylaminosulfonyl)amino)-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoic acid

[0195] In a 50 mL single-necked flask, add the product from the previous step (1.0 g), tetrahydrofuran (20 mL), and water (5 mL) in sequence and stir until dissolved. Then, add lithium hydroxide monohydrate (130 mg) to the system and continue stirring at room temperature for 3 h. Add hydrochloric acid (1 M) to the reaction solution to adjust the pH to 5-6. Concentrate under reduced pressure to remove tetrahydrofuran. Dilute the concentrate with water, filter, and collect the filter cake to obtain 750 mg of a brown solid. ESI-MS: [MH] - =623.0.

[0196] Step 3: Preparation of N'-allyl-3,4-difluoro-5-((3-fluoro-2-((N-ethylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzohydrazide

[0197] In a 100 mL single-necked flask, add the product from the previous step (750 mg), N,N-dimethylformamide (8 mL), 1-hydroxybenzotriazole (240 mg), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (690 mg), and stir the reaction at room temperature for 2 hours. Add allylhydrazine hydrochloride (260 mg) and N,N-diisopropylethylamine (470 mg) to the system, and continue stirring the reaction at room temperature for 1 hour. The reaction solution is slowly added to a saturated sodium bicarbonate aqueous solution, extracted with ethyl acetate, and the organic phases are combined, dried, and concentrated. The concentrate is purified by silica gel column chromatography to obtain the product. ESI-MS: [M+H] + =679.1.

[0198] Example 13

[0199] Referring to Example 12, N-methyl-d3-2-oxoxazoline-3-sulfonamide was substituted for N-ethyl-2-oxoxazoline-3-sulfonamide to synthesize compound 4-8. ESI-MS: [M+H] + =668.1.

[0200] Example 14

[0201] Refer to the synthesis method of Intermediate 2 to synthesize (allyl-1,1-d2)hydrazine hydrochloride, and then refer to Example 7 to synthesize compound 4-9. ESI-MS: [M+H] + =667.1.

[0202] Example 15

[0203] Step 1: Preparation of tert-butyl 2-(prop-2-yn-1-yl)hydrazine-1-carboxylate

[0204] To a 250 mL single-necked flask, add tert-butyl hydrazinecarboxylate (10.0 g), dimethyl sulfoxide (50 mL), triethylamine (6.8 g), and propargyl bromide (8.0 g). After nitrogen replacement, stir at room temperature for 3 h. Add water to the reaction solution, extract with ethyl acetate, and combine the organic phases, dry, and concentrate. The concentrate is purified by silica gel column chromatography to yield 1.5 g of a white solid.

[0205] Step 2: Preparation of prop-2-yn-1-ylhydrazine hydrochloride

[0206] The product from the previous step (2.2 g) and tetrahydrofuran (6 mL) were added to a 100 mL single-necked flask. Concentrated hydrochloric acid (6 mL) was slowly added dropwise with stirring at room temperature. After completion of the addition, the system was stirred at room temperature for 3 h. The reaction solution was concentrated to obtain 1.0 g of a yellow oil. ESI-MS: [M+H] + =71.1.

[0207] Step 3: Preparation of N'-(prop-2-yn-1-yl)-3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzohydrazide

[0208] To a 100 mL single-necked flask, 3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzoic acid (1.0 g) and N,N-dimethylformamide (20 mL) were added. The mixture was cooled to 0°C in an ice bath, followed by the addition of 1-hydroxybenzotriazole (266 mg) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (629 mg). The mixture was stirred for 5 minutes and then allowed to warm to room temperature for 2 hours. Prop-2-yn-1-ylhydrazine hydrochloride (351 mg) and N,N-diisopropylethylamine (793 mg) were added, and the mixture was stirred at room temperature for another 2 hours. The reaction mixture was slowly added to a saturated sodium bicarbonate solution, extracted with ethyl acetate, and the organic phases were combined, dried, and concentrated. The concentrate was purified by silica gel column chromatography to obtain 200 mg of an off-white solid. ESI-MS:[M+H] + =663.0.

[0209] Example 16

[0210] Step 1: Preparation of tert-butyl 2-methyl-2-(prop-2-yn-1-yl)hydrazine-1-carboxylate

[0211] To a 50 mL single-necked flask, add tert-butyl 2-(prop-2-yn-1-yl)hydrazine-1-carboxylate (1.4 g), potassium carbonate (1.7 g), iodomethane (1.8 g), and N,N-dimethylformamide (20 mL). Heat to 30°C and stir for 2 h. Pour the reaction solution into water and extract with ethyl acetate. The organic phases are combined, dried, and concentrated to yield 1.0 g of a colorless liquid.

[0212] Step 2: Preparation of 1-methyl-1-(prop-2-yn-1-yl)hydrazine hydrochloride

[0213] The product from the previous step (1.0 g) and a 4M solution of hydrogen chloride in ethyl acetate (10 mL) were added to a 50 mL single-necked flask and stirred at room temperature for 2 h. The reaction solution was concentrated to obtain 410 mg of a white solid. ESI-MS: [M+H] + =85.1.

[0214] Step 3: Preparation of N'-methyl-N'-(prop-2-yn-1-yl)-3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzohydrazide

[0215] To a 100 mL single-necked flask, add 3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzoic acid (400 mg) and N,N-dimethylformamide (10 mL), and cool to 0°C in an ice bath. Then, add 1-hydroxybenzotriazole (107 mg) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (252 mg), stir for 5 minutes, and then warm to room temperature for 2 hours. Then, add 1-methyl-1-(prop-2-yn-1-yl)hydrazine hydrochloride (95 mg) and N,N-diisopropylethylamine (254 mg), and continue stirring at room temperature for 2 hours. The reaction solution is slowly added to a saturated sodium bicarbonate solution, extracted with ethyl acetate, and the organic phases are combined, dried, and concentrated. The concentrate was purified by silica gel column chromatography to obtain 150 mg of an off-white solid. 1H NMR (400MHz, DMSO-d6): δ10.43(s,1H),9.62(s,1H),8.55(s,1H),8.02(d,J=4 .4Hz,1H),7.57(dd,J=10.8,2.0Hz,1H),7.41(d,J=7.2Hz,1H),7.35(dd,J=8. 4,0.8Hz,1H),7.04(brs,1H),6.93(brs,1H),6.67-6.61(m,1H),4.07(s,2H), 3.56(d,J=2.0Hz,2H),3.14(t,J=2.0Hz,1H),2.54-2.51(m,6H).ESI-MS:[M+H] + =677.1.

[0216] Example 17

[0217] Step 1: Preparation of tert-butyl 2-(3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzoyl)-1-methylhydrazine-1-carboxylate

[0218] To the reaction flask, 3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzoic acid (500 mg), 1-tert-butoxycarbonyl-1-methylhydrazine (145 mg), and N,N-dimethylformamide (10 mL) were added and cooled to 0°C in an ice bath. Subsequently, O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate (395 mg) and N,N-diisopropylethylamine (265 mg) were added. After the addition, the mixture was allowed to warm to room temperature and stirred for 2 hours. The reaction solution was slowly added to a saturated sodium bicarbonate solution, extracted with ethyl acetate, and the organic phases were combined, dried, and concentrated. The concentrate was purified by silica gel column chromatography to obtain 442 mg of the product.

[0219] Step 2: Preparation of N'-methyl-3,4-difluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzohydrazide

[0220] The product from the previous step (440 mg), dichloromethane (10 mL), and trifluoroacetic acid (2 mL) were added to the reaction flask and stirred at room temperature for 2 h. The reaction solution was slowly added to a cold aqueous sodium bicarbonate solution and extracted with dichloromethane. The organic phase was collected, washed with a saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The concentrate was purified by silica gel column chromatography to obtain 263 mg of the product. ESI-MS: [M+H]+ =639.0.

[0221] Example 18

[0222] Step 1: Preparation of 2,4-difluoro-5-vinylbenzoic acid

[0223] In a 3L three-necked flask, 2,4-difluoro-5-iodobenzoic acid (10.0 g), potassium vinyl trifluoroborate (5.7 g), potassium phosphate (11.2 g), methanol (200 mL), water (100 mL) and 1,1'-bis(diphenylphosphino)ferrocene palladium dichloride (2.6 g) were added in sequence. The system was replaced with a nitrogen atmosphere and heated to 60°C for 21 hours. After the system was cooled to room temperature, it was filtered through celite, the filtrate was collected, concentrated, and 1M HCl was added to adjust the pH of the system to 4-5. The system was then extracted with tertiary methyl ether, the organic phases were combined, washed with saturated brine, dried and concentrated. The concentrate was purified by slurrying with a mixed solvent of petroleum ether / dichloromethane (v / v=9 / 1) to obtain 6.3 g of a brown solid. ESI-MS: [MH] - =183.0.

[0224] Step 2: Preparation of 4-fluoro-2-((2-fluoro-4-iodophenyl)amino)-5-vinylbenzoic acid

[0225] In a 500mL three-necked flask, add the product of the previous step (6.2g), 2-fluoro-4-iodoaniline (8.0g) and tetrahydrofuran (120mL), then replace the system with a nitrogen environment and cool to -70~-80℃. Slowly add bistrimethylsilylamine lithium (1M, 101mL) to the system, ensuring that the system temperature does not exceed -70℃. After the dropwise addition, slowly return the system to room temperature and keep the reaction for 30min. Then add hydrochloric acid (1M) to the system to adjust the pH of the system to 1~2. Use tert-butyl methyl ether to extract the system, combine the organic phases, wash with saturated brine, dry and concentrate. Slowly add tert-butyl methyl ether and petroleum ether to the concentrate, stir at room temperature for 30min, filter to obtain 11.0g of light brown solid.

[0226] Step 3: Preparation of methyl 4-fluoro-2-((2-fluoro-4-iodophenyl)amino)-5-vinylbenzoate

[0227] The product from the previous step (11.0 g) and 110 mL of N,N-dimethylformamide were added to a 250 mL three-necked flask and stirred at room temperature until dissolved. Potassium carbonate (5.7 g) was added to the system, the temperature was raised to 50°C and stirred for 2 h, then the temperature was lowered to 25°C. Methyl iodide (5.8 g) was added and the reaction was allowed to react for 3 h. The reaction solution was added to an aqueous ammonium chloride solution and extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried, and concentrated. Purification by methanol slurrying gave 9.3 g of a light brown solid. 1 H NMR (400MHz, DMSO-d6): δ9.51(s,1H),8.12(d,J=8.8Hz,1H),7.76(dd,J1=10.0Hz,J2=1.2Hz,1H),7.57(d,J=8.4Hz,1H ),7.34(t,J=8.4Hz,1H),6.78-6.67(m,2H),5.76(d,J=17.6Hz,1H),5.31(d,J=11.6Hz,1H),3.88(s,3H).ESI-MS:[M+H] + =416.0.

[0228] Step 4: Preparation of methyl 4-fluoro-2-((2-fluoro-4-iodophenyl)amino)-5-formylbenzoate

[0229] In a 500mL three-necked flask, add the product from the previous step (9.3g) and tetrahydrofuran (150mL), stir at room temperature until dissolved, and cool to 0°C. Add potassium osmate dihydrate (413mg) to the system and stir for 5min. Add an aqueous solution (150mL) of sodium periodate (14.4g) to the system and stir for 5min. Then add 2,4-lutidine (2.4g) to the system and react at room temperature for 8h. The reaction solution is slowly added to a 20% sodium sulfite solution, extracted with ethyl acetate, and the organic phases are combined, dried and concentrated, and purified by ethanol slurrying to obtain 5.6g of light brown solid. 1 H NMR (400MHz, DMSO-d6): δ9.95-9.22(m,2H),8.42(d,J=7.2Hz,1H),7.83(d,J=9.6Hz,1H),7.6 4(d,J=7.6Hz,1H),7.35(t,J=8.0Hz,1H),6.64(d,J=13.2Hz,1H),3.91(s,3H).ESI-MS:[M+H] + =418.0.

[0230] Step 5: Preparation of methyl (E)-4-fluoro-2-((2-fluoro-4-iodophenyl)amino)-5-((2-toluenesulfonylhydrazide)methyl)benzoate

[0231] To a 250 mL three-necked flask, add the product from the previous step (5.6 g), p-methylbenzenesulfonylhydrazide (2.5 g), and ethanol (120 mL). Heat the system to 50°C and react for 5 h. Cool the system to 0°C in an ice bath to precipitate a solid. Filter the solid, and wash the filter cake with ethanol to obtain 7.0 g of a gray solid. 1 H NMR (400MHz, DMSO-d6): δ11.5(s,1H),9.60(s,1H),8.24(d,J=8.4Hz,1H),7.95(s,1H),7.77(dd,J1=10.0Hz,J2=1.6Hz,1H),7.73(d,J=8.4H z,2H),7.58(d,J=8.4Hz,1H),7.42(d,J=8.4Hz,2H),7.32(t,J=8.4Hz,1H),6.68(d,J=12.8Hz,1H),3.91(s,3H),2.37(s,3H).ESI-MS:[M+H] + =586.0.

[0232] Step 6: Preparation of methyl 5-((2-((2,4-dimethoxybenzyl)amino)-3-fluoropyridin-4-yl)methyl)-4-fluoro-2-((2-fluoro-4-iodophenyl)amino)benzoate

[0233] To a 500 mL single-necked flask, add (2-((2,4-dimethoxybenzyl)amino)-3-fluoropyridin-4-yl)boric acid (5.1 g), the product from the previous step (7.0 g), potassium carbonate (2.8 g), and toluene (300 mL). Replace the atmosphere with nitrogen. Heat the system to 100°C and react for 14 hours. After cooling to room temperature, filter, remove the solvent, and purify the crude product by silica gel column chromatography to obtain 6.0 g of a pale yellow solid. 1 H NMR (400MHz, DMSO-d6): δ9.44(s,1H),7.92(d,J=8.8Hz,1H),7.74(dd,J1=10.4Hz,J2=1.6Hz,1H), 7.65(d,J=5.2Hz,1H),7.54(dd,J1=8.4Hz,J2=1.2Hz,1H),7.33(t,J=8.4Hz,1H),7.02(d,J=8.4Hz ,1H),6.84-6.77(m,2H),6.52(d,J=2.4Hz,1H),6.41(dd,J1=8.4Hz,J2=2.0Hz,1H),6.31(t,J=4.8 Hz,1H),4.43(d,J=6.0Hz,2H).3.87(s,2H),3.85(s,3H),3.79(s,3H),3.71(s,3H).ESI-MS:[M+H]+ =664.1.

[0234] Step 7: Preparation of methyl 5-((2-amino-3-fluoropyridin-4-yl)methyl)-4-fluoro-2-((2-fluoro-4-iodophenyl)amino)benzoate

[0235] In a 500mL single-necked bottle, the product of the previous step (5.9g) and dichloromethane (118mL) were added and stirred at room temperature until dissolved. The system was then placed in an ice bath and trifluoroacetic acid (59mL) was slowly added. After the addition, the system was warmed to room temperature and stirred for 1.5h. The system was slowly added to a saturated aqueous sodium bicarbonate 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 3.9g of a light yellow solid. ESI-MS: [M+H] + =514.0.

[0236] Step 8: Preparation of methyl 4-fluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzoate

[0237] To a 100mL three-necked flask, add the product from the previous step (1.0g) and N,N-dimethylformamide (20mL) and stir until dissolved. Pyridine (1.6g) is then added and the temperature is lowered to -15°C. Methylaminosulfonyl chloride (0.8g) is then added dropwise. Stir the reaction at -15°C for 1 hour. Pour the mixture into water and extract with ethyl acetate. The organic phases are combined, dried, and concentrated to yield 1.1g of a brown solid.

[0238] Step 9: Preparation of 4-fluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzoic acid

[0239] In a 1L single-necked flask, the product from the previous step (1.1 g), tetrahydrofuran (17 mL), and water (9 mL) were added sequentially and stirred in an ice bath. Lithium hydroxide monohydrate (0.2 g) was slowly added to the system. After addition, the mixture was warmed to room temperature and stirred for 16 hours. Aqueous hydrochloric acid (1 N) was then added to the system to adjust the pH to 1-2 and stirred for 5 minutes. Extraction with ethyl acetate was performed, and the organic phases were combined, dried, and concentrated to yield 1.0 g of a brown solid.

[0240] Step 10: Preparation of tert-butyl 2-(4-fluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzoyl)hydrazine-1-carboxylate

[0241] In a 100 mL single-necked flask, the product from the previous step (1.0 g) and N,N-dimethylformamide (20 mL) were added. 1-Hydroxybenzotriazole (0.5 g) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.7 g) were then added to the system and stirred at room temperature for 1 hour. Tert-butyl carbazate (0.5 g) and N,N-diisopropylethylamine (0.9 g) were then added to the system and stirred at room temperature for 2 hours. Water was slowly added to the reaction system, extracted with ethyl acetate, and the organic phases were combined, dried, and concentrated. Purification by silica gel column chromatography yielded 800 mg of a brown solid. 1 H NMR (400MHz, DMSO-d6): δ10.28(s,1H),10.25(s,1H),9.69(s,1H),8.91(s,1H),8.00(d,J=4.8Hz 1H),7.76(s,1H),7.69(dd,J1=10.8Hz,J2=1.2Hz,1H),7.51(d,J=8.4Hz,1H),7.30( t,J=8.4Hz,1H),6.92-6.88(m,3H),3.94(s,2H),2.53(d,J=4.8Hz,3H),1.43(s,9H).

[0242] Step 11: Preparation of 4-fluoro-5-((3-fluoro-2-((N-methylaminosulfonyl)amino)pyridin-4-yl)methyl)-2-((2-fluoro-4-iodophenyl)amino)benzohydrazide

[0243] In a 100mL single-necked bottle, the product of the previous step (800mg) and ethyl acetate (40mL) were added, and the system was stirred at room temperature for 5min. Subsequently, a solution of hydrogen chloride in ethyl acetate (4M, 8mL) was slowly added dropwise to the system, and the reaction was continued to stir at room temperature for 4h. The reaction system was added dropwise to a saturated aqueous sodium bicarbonate solution and the system was extracted with ethyl acetate. The organic phase was dried, filtered, and concentrated to give 640mg of the product as a white solid. 1 H NMR (400MHz, DMSO-d6): δ10.38 (s, 1H), 9.93-9.91 (m, 2H), 7.99 (d, J = 4.8Hz 1H),7.70-7.65(m,2H),7.50-7.47(m,1H),7.30(t,J=8.4Hz,1H),6.99- 6.90(m,3H),4.55(s,2H),3.92(s,2H),2.52-2.51(m,3H).ESI-MS:[M+H] + =607.0.

[0244] Preparation of control compounds

[0245] Reference compounds TX-1 and TX-2 were prepared from reference document CN114929669, which are compounds A-13 and A-7 in the reference document, respectively, and have the following structures:

[0246] 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.

[0247] Biological testing

[0248] 1. Proliferation inhibition activity test of pancreatic cancer MIA PaCa-2 cells

[0249] Cell proliferation activity was assessed using the CellTiter-3D-Glo assay. The assay involved digesting MIA PaCa-2 cells in the logarithmic growth phase, diluting them to an appropriate cell density using DMEM complete medium containing 10% FBS, 2.5% HS, and 1% PS, and seeding them in a low-adsorption 96-well U-bottom plate in a 5% CO2 incubator. A series of concentration gradients of the test compound were added, and the cells were incubated in a 5% CO2 incubator. Following incubation, the cells were lysed and assayed using the CellTiter-3D-Glo assay. Chemiluminescence values ​​were measured using a microplate reader, and data were fitted using Graphpad Prism software to calculate the IC values ​​of the test compounds. 50 The test results are shown in the table below.

[0250] “A” indicates inhibitory activity IC 50 <20 nM, "B" indicates inhibitory activity 20 nM ≤ IC 50 <50nM, “C” represents the inhibitory activity IC 50 ≥50nM.

[0251] The test results show that the compound of the present invention exhibits significant cell proliferation inhibitory activity on pancreatic cancer MIA PaCa-2 cells.

[0252] 2. Pharmacodynamics Study in Subcutaneous Pancreatic Cancer ASPC-1 Cell Xenograft Model

[0253] Animal species and number: BALB / c nude mice, 6 per group;

[0254] Test sample: Compound 4-1;

[0255] Experimental groups: blank solvent control group, compound 4-1 (0.2 mg / kg, QD×21 days);

[0256] Animal model establishment: Pancreatic cancer ASPC-1 cells in the logarithmic growth phase were cultured in vitro and collected, then subcutaneously inoculated into the right dorsal region of Balb / c nude mice. When tumors grew to 200-300 mm3, the tumor-bearing mice were randomly divided into groups based on tumor volume. Each group of animals was then dosed, and the day of the first dose was defined as experimental day 1.

[0257] Administration route and frequency: oral gavage, once a day for 21 consecutive days;

[0258] General status observation: Observation time and frequency: once a day; Observation indicators or contents: including but not limited to the animal's drug administration site, external signs, general behavioral activities, mental state, death, and other abnormal manifestations.

[0259] Tumor volume calculation: V = 0.5a × b 2 , where a is the long diameter of the tumor and b is the short diameter of the tumor. The tumor growth inhibition rate (TGI) (%) was used to evaluate the anti-tumor efficacy of the compound. TGI (%) = [1 - (average tumor volume of the treatment group at the end of dosing - average tumor volume of the treatment group at the start of dosing) / (average tumor volume of the control group at the end of dosing - average tumor volume of the control group at the start of dosing)] × 100%.

[0260] “+” indicates a tumor inhibition rate of <60%; “++” indicates a tumor inhibition rate of 60% to 100%; and “+++” indicates a tumor inhibition rate of >100%.

[0261] The test results showed that compound 4-1 significantly inhibited the growth of pancreatic cancer ASPC-1 cell subcutaneous transplanted tumors in nude mice. During the dosing period, the animals in the treatment group maintained normal diet, body weight, and activity, demonstrating a good safety profile.

[0262] 3. Solubility test

[0263] Test compound: Compound 4-1;

[0264] Test procedure: In a 25 mL stoppered colorimetric tube, add 5 mL of pH 1.2 hydrochloric acid solution, then add compound 4-1, stir in a 37°C water bath, filter through a polyethersulfone filter membrane, and determine the concentration of the filtrate by HPLC. The results are shown in the table below.

[0265] “+” indicates solubility <10 μg / ml; “++” indicates solubility of 10-30 μg / ml; “+++” indicates solubility >30 μg / ml.

[0266] The test results showed that compound 4-1 had good solubility in pH 1.2 hydrochloric acid solution.

[0267] 4. Proliferation Inhibitory Activity Test of Pancreatic Cancer MIA PaCa-2 Cells

[0268] Referring to Biological Test Example 1, the proliferation inhibitory activity of the compounds of the present invention on pancreatic cancer MIA PaCa-2 cells was tested. The test results are shown in the following table. Wherein, "A" represents the inhibitory activity IC 50 <20 nM, "B" indicates inhibitory activity 20 nM ≤ IC 50 <50nM, “C” represents the inhibitory activity IC 50 ≥50nM.

[0269] The test results show that the compound of the present invention has significant cell proliferation inhibitory activity on pancreatic cancer MIA PaCa-2 cells.

[0270] 5. Proliferation Inhibitory Activity Test of Colorectal Cancer SW480 Cells

[0271] Cell proliferation activity was assessed using the CellTiter-3D-Glo assay. The assay procedure was as follows: SW480 cells in the logarithmic growth phase were digested, diluted to an appropriate cell density using DMEM complete medium supplemented with 10% FBS and 1% PS, and seeded into low-adsorption 96-well U-bottom plates in a 5% CO2 incubator. A series of concentration gradients of the test compound were added and incubated in a 5% CO2 incubator. Following incubation, the cells were lysed and assayed using the CellTiter-3D-Glo assay. Chemiluminescence values ​​were measured using a microplate reader, and data were fitted using Graphpad Prism software to calculate the IC values ​​of the test compounds. 50 The test results are shown in the following table:

[0272] The test results show that the compound of the present invention exhibits significant cell proliferation inhibitory activity on colorectal cancer SW480 cells and is significantly better than the control compound TX-1.

[0273] 6. Proliferation inhibition activity test of lung cancer NCI-H358 cells

[0274] Cell proliferation activity was assessed using the CellTiter-3D-Glo assay. The assay procedure was as follows: NCI-H358 cells in the logarithmic growth phase were digested and seeded into low-adsorption 96-well U-bottom plates at the appropriate density. The cells were then cultured in a cell culture incubator. A concentration gradient of the test substance was added to the experimental wells, while control medium was added to the control wells. The cells were then incubated in a 5% CO2 incubator. After incubation, the cells were lysed and assayed using the CellTiter-3D-Glo assay. Chemiluminescence was measured using a microplate reader. Data were fitted using Graphpad Prism software to calculate the IC values ​​of the test substances. 50The test results are shown in the following table:

[0275] The test results show that the compound of the present invention exhibits significant cell proliferation inhibitory activity on lung cancer NCI-H358 cells and is significantly better than the comparative compound TX-2.

[0276] 7. Pharmacodynamics Study in Subcutaneous Xenograft Model of Colorectal Cancer Cell SW480

[0277] Animal species and number: BALB / c nude mice, 6 per group;

[0278] Test samples: Compound 3-9, Compound 4-1;

[0279] Test groups: blank solvent control group, compound 3-9 (0.2 mg / kg, QD×24 days), compound 4-1 (0.2 mg / kg, QD×24 days);

[0280] Animal model establishment: SW480 tumor cells in the logarithmic growth phase were cultured in vitro and collected, and subcutaneously inoculated into the right back of Balb / c Nude mice. 3 Around 24 hours after the start of the experiment, the tumor-bearing mice were randomly divided into groups. Subsequently, each group of animals was dosed, and the day of the first dose was defined as the first day of the experiment.

[0281] Administration route and frequency: oral gavage, once a day for 24 consecutive days;

[0282] General status observation: Observe once a day. The observation contents include but are not limited to the animal's drug administration site, appearance signs, general behavior activities, mental state, death, and other abnormal manifestations.

[0283] Tumor volume calculation: V = 0.5a × b 2 , where a is the long diameter of the tumor and b is the short diameter of the tumor. The tumor growth inhibition rate (TGI) (%) was used to evaluate the anti-tumor efficacy of the compound. TGI (%) = [1 - (average tumor volume of the treatment group at the end of dosing - average tumor volume of the treatment group at the start of dosing) / (average tumor volume of the control group at the end of dosing - average tumor volume of the control group at the start of dosing)] × 100%.

[0284] “+” indicates a tumor inhibition rate of <60%; “++” indicates a tumor inhibition rate of 60% to 90%; and “+++” indicates a tumor inhibition rate of >90%.

[0285] The experiments showed that compounds 3-9 and 4-1 significantly inhibited the growth of subcutaneous transplanted tumors of colorectal cancer SW480 cells in nude mice. During the dosing period, the animals in each treatment group maintained normal diet, body weight, and activity, demonstrating a good safety profile.

Claims

1. A compound represented by formula (I), its stereoisomers, tautomers or pharmaceutically acceptable salts thereof, Among them, Z is selected from oxygen or sulfur; U1 is selected from nitrogen or CR 5d ; U2 is selected from nitrogen or CR 5a ; U3 is selected from nitrogen or CR 5b ; U4 is selected from nitrogen or CR 5c ; Ring B is selected from substituted or unsubstituted aryl or heteroaryl; R A selected from -SO2NR a R b , -NR a SO2NR b R c , -NR a SO2R b , -C(O)NR a R b , -NR a C(O)NR b R c or -NR a C(O)R b ; R 2a ,R 2b ,R 2c each independently selected from hydrogen, deuterium, -OR a ,-SR a ,C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-6 cycloalkyl, 3- to 10-membered heterocyclic group, C 6-10 aryl or 5- to 10-membered heteroaryl, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic group, aryl, heteroaryl may be substituted by one or more substituents independently selected from deuterium, halogen, aldehyde group, cyano group, amino group, hydroxyl group, oxo, C 1-6 alkyl, C 1-6 alkoxy, C 2-6 alkenyl, C 2-6 alkynyl, C 3-6 cycloalkyl, 3- to 10-membered heterocyclic group, C 6-10 aryl or 5- to 10-membered heteroaryl; R 3a ,R 3b ,R 5a ,R 5b ,R 5c ,R 5d each independently selected from hydrogen, deuterium, halogen, aldehyde group, cyano group, amino group, hydroxyl group, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 2-6 alkenyl, C 2-6 alkynyl or C 3-6 cycloalkyl; or two adjacent substituents may together form a substituted or unsubstituted 3- to 10-membered ring structure; X is selected from -O-, -CR a R b or -NR a ; Ring A is selected from substituted or unsubstituted aryl or heteroaryl; R a ,R b ,R c each independently selected from hydrogen, deuterium, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 alkoxy, C 2-6 alkenyl, C 2-6 deuterated alkenyl, C 2-6 alkynyl, C 2-6 deuterated alkynyl, C 3-6 cycloalkyl, 3- to 10-membered heterocyclic group, C 6-10 aryl or 5- to 10-membered heteroaryl; n is 1, 2, 3 or 4.

2. A compound of formula (II), its stereoisomers, tautomers or pharmaceutically acceptable salts thereof, Among them, U1 is selected from nitrogen or CR 5d ; R A selected from -SO2NR a R b , -NR a SO2NR b R c , -NR a SO2R b , -C(O)NR a R b , -NR a C(O)NR b R c or -NR a C(O)R b ; R a ,R b ,R c are each independently selected from hydrogen, deuterium, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 alkoxy, C 2-6 alkenyl, C 2-6 deuterated alkenyl, C 2-6 alkynyl, C 2-6 deuterated alkynyl, C 3-6 cycloalkyl, 3- to 10-membered heterocyclic group, C 6-10 aryl or 5- to 10-membered heteroaryl; R 2a ,R 2b ,R 2c each independently selected from hydrogen, deuterium, -OR a , -SR a , C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-6 cycloalkyl, 3- to 10-membered heterocyclic group, C 6-10 aryl or 5- to 10-membered heteroaryl, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic group, aryl, heteroaryl may be substituted by one or more substituents independently selected from deuterium, halogen, aldehyde group, cyano group, amino group, hydroxyl group, oxo, C 1-6 alkyl, C 1-6 alkoxy, C 2-6 alkenyl, C 2-6 alkynyl, C 3-6 cycloalkyl, 3- to 10-membered heterocyclic group, C 6-10 aryl or 5- to 10-membered heteroaryl; 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, aldehyde group, cyano group, amino group, hydroxyl group, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 2-6 alkenyl, C 2-6 alkynyl or C 3-6 cycloalkyl; or two adjacent substituents may together form a substituted or unsubstituted 3- to 10-membered ring structure.

3. A compound represented by formula (III), its stereoisomers, tautomers or pharmaceutically acceptable salts thereof, Among them, U1 is selected from nitrogen or Cr 5d ; R 2a and R 2b are each independently selected from hydrogen, deuterium, -OR a , -SR a , C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-6 cycloalkyl, 3-10 membered heterocyclic group, C 6-10 aryl or 5-10 membered heteroaryl, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic group, aryl, heteroaryl may be substituted by one or more substituents selected from deuterium, halogen, aldehyde group, cyano group, amino group, hydroxyl group, oxo, C 1-6 alkyl, C 1-6 alkoxy, C 2-6 alkenyl, C 2-6 alkynyl, C 3-6 cycloalkyl, 3-10 membered heterocyclic group, C 6-10 aryl or 5-10 membered heteroaryl; R a selected from hydrogen, deuterium, C 1-6 alkyl, C 1-6 deuterated alkyl, C 2-6 alkenyl, C 2-6 deuterated alkenyl, C 2-6 alkynyl, C 2-6 deuterated alkynyl or C 3-6 cycloalkyl; R 3a ,R 3b each independently selected from hydrogen, deuterium, halogen or C 1-6 alkyl; 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 independently selected from hydrogen, deuterium, halogen, aldehyde group, cyano group, amino group, hydroxyl group, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 2-6 alkenyl, C 2-6 alkynyl or C 3-6 cycloalkyl; or two adjacent substituents may together form a substituted or unsubstituted 3- to 10-membered ring structure; R7 is selected from hydrogen, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 3-6 cycloalkyl, C 2-6 alkenyl or C 2-6 alkynyl.

4. The following compounds, their stereoisomers, tautomers or pharmaceutically acceptable salts, 5. A compound of formula (IV-I), its stereoisomers, tautomers or pharmaceutically acceptable salts thereof, Among them, U1 is selected from nitrogen or CR 5d ; R 2b ,R 3a ,R 3b is selected from hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 deuterated alkyl or C 1-6 haloalkyl; 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 independently selected from hydrogen, deuterium, halogen, aldehyde group, cyano group, amino group, hydroxyl group, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 2-6 alkenyl, C 2-6 alkynyl or C 3-6 cycloalkyl; or two adjacent substituents may together form a substituted or unsubstituted 3- to 10-membered cyclic structure; R7 is selected from hydrogen, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 3-6 cycloalkyl, C 2-6 alkenyl or C 2-6 alkynyl; R 8a ,R 8b ,R 8c ,R 8d ,R 8e each independently selected from hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl or C 3-6 cycloalkyl; n is 1, 2, 3 or 4.

6. A compound represented by formula (IV-II), its stereoisomers, tautomers or pharmaceutically acceptable salts thereof, Among them, U1 is selected from nitrogen or CH; R 2b selected from hydrogen, deuterium or C 1-6 alkyl; R 4a ,R 4c each independently selected from hydrogen, deuterium, halogen, aldehyde group, hydroxyl group, C 1-6 alkyl, C 1-6 alkoxy, C 2-6 alkenyl, C 2-6 alkynyl or C 3-6 cycloalkyl; R 5c ,R 6b ,R 6c are each independently selected from hydrogen, deuterium, a halogen or a C 1-6 alkyl group; 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 ,R 8d ,R 8e each independently selected from hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 deuterated alkyl or C 1-6 haloalkyl; n is 1, 2, 3 or 4.

7. A compound represented by formula (IV-III), its stereoisomers, tautomers or pharmaceutically acceptable salts thereof, Among them, R 2b selected from hydrogen or C 1-6 alkyl group; R 4c selected from halogen, aldehyde group, C 1-6 alkyl group, C 1-6 alkoxy group, C 2-6 alkenyl group, C 2-6 alkynyl group or C 3-6 cycloalkyl group; R7 is selected from C 1-6 alkyl, C 1-6 deuterated alkyl or C 3-6 cycloalkyl; R 8a ,R 8b ,R 8c ,R 8d ,R 8e Each independently selected from hydrogen, deuterium or C 1-6 alkyl; n is 1, 2, 3 or 4.

8. The following compounds, their stereoisomers, tautomers or pharmaceutically acceptable salts thereof, 9. A compound represented by formula (V), its stereoisomers, tautomers or pharmaceutically acceptable salts thereof, Among them, U1 is selected from nitrogen or CR 5d ; R 2b ,R 3a ,R 3b is selected from hydrogen, deuterium, a halogen, C 1-6 alkyl, C 1-6 deuterated alkyl or C 1-6 haloalkyl; 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 independently selected from hydrogen, deuterium, halogen, aldehyde group, cyano group, amino group, hydroxyl group, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 2-6 alkenyl, C 2-6 alkynyl or C 3-6 cycloalkyl; or two adjacent substituents may together form a substituted or unsubstituted 3- to 10-membered cyclic structure; R7 is selected from hydrogen, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 3-6 cycloalkyl, C 2-6 alkenyl or C 2-6 alkynyl; R 8a ,R 8b ,R 8c are each independently selected from hydrogen, deuterium, a halogen, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl or C 3-6 cycloalkyl; n is 1, 2, 3 or 4.

10. The following compounds, their stereoisomers, tautomers or pharmaceutically acceptable salts, 11. A pharmaceutical composition comprising a therapeutically effective dose of a compound according to any one of claims 1 to 10, or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

12. Use of a compound according to any one of claims 1 to 10, a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 11, in the preparation of a medicament for preventing and / or treating tumors.

13. Use of a compound according to any one of claims 1 to 10, a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 11, in the preparation of a medicament for preventing and / or treating a disease related to the MAPK pathway, wherein the disease particularly refers to a tumor.

14. Use according to claim 12 or 13, wherein the tumors include renal cancer, liver cancer, breast cancer, pancreatic cancer, prostate cancer, melanoma, leukemia, malignant lymphoma, ovarian cancer, head and neck cancer, lung cancer, colorectal cancer, bladder cancer, uterine cancer, etc.