Compounds with ERK kinase inhibitory activity and uses thereof

Abstract

A compound of formula (I): #imgabs0# wherein the variables are as defined in the specification. The compound is an inhibitor of ERK kinases, such as ERK1 and / or ERK2 kinases. Also disclosed are uses and methods for preparing the compound, and pharmaceutical compositions comprising the compound.

Description

Technical Field

[0001] The present invention belongs to the field of medicinal chemistry. Specifically, the present invention relates to novel compounds or pharmaceutically acceptable salts thereof, and pharmaceutical compositions containing the compounds or pharmaceutically acceptable salts thereof, which are useful as modulators of the extracellular signal-regulated kinase (ERK) pathway, particularly as inhibitors of ERK kinases, such as ERK1 and / or ERK2 kinases. Background Art

[0002] The Ras-Raf-MEK-ERK pathway is a mitogen-activated protein kinase (MAPK) signaling pathway that regulates numerous cell functions, including proliferation, differentiation, and apoptosis. Mutations in this pathway are present in over one-third of all human cancers, making node proteins in this pathway a hot topic in the development of targeted cancer drugs in recent years. The specific B-Raf inhibitors vemurafenib and dabrafenib were approved by the US FDA for the treatment of melanoma in 2011 and 2013, respectively. The MEK1 / 2 inhibitor trametinib was also approved by the US FDA for the treatment of melanoma in 2013. In 2015, the combination of vemurafenib and the MEK inhibitor cobimetinib was approved by the US FDA for the treatment of melanoma harboring B-Raf V600E or V600K mutations. In 2017, the US FDA also approved the combination of dabrafenib and trametinib for the treatment of B-Raf V600E mutant non-small cell lung cancer. However, inhibiting these upstream pathway nodes has its limitations. Tumors can rapidly develop resistance to B-Raf and MEK inhibitors. The mechanisms of resistance include point mutations, changes in protein polymerization, and alterations in protein peptide length, posing a significant challenge to the development of next-generation drugs targeting Raf and MEK. Furthermore, as a key terminal node in the MAPK pathway, activated ERK can transmit extracellular signals to the nucleus, promoting the phosphorylation of cytoplasmic target proteins or regulating the activity of other protein kinases, thereby regulating gene expression. Its importance in the development of oncology drugs is undeniable. This is particularly true given that most current MAPK upstream targeted therapies ultimately develop resistance. ERK inhibitors may offer a more effective therapeutic approach because they are less susceptible to acquired resistance. Since its discovery in the 1990s, ERK has been extensively and intensively studied, but to date, no ERK inhibitors have been approved for marketing. Currently, the leading candidate internationally is the highly selective ERK inhibitor BVD-523 (ulixertinib), currently in Phase 2 clinical trials. Reports in early 2017 demonstrated that a 600mg twice-daily dose of BVD-523 had an acceptable safety profile and produced durable responses in patients with NRAS-mutant melanoma, and BRAF V600 and non-V600 mutant solid tumors (including melanoma, glioblastoma multiforme, brain metastases, gallbladder adenocarcinoma, and head and neck cancer). These data further support the clinical development of ERK inhibitors.

[0003] In summary, it can be expected that ERK inhibitors will have broad prospects in the field of tumors as single drugs or combined drugs, and the field is in urgent need of developing new ERK inhibitors. Patent application WO2017 / 114510A1 discloses a series of ERK inhibitors, but the inventors of this application found that some compounds in the patent application, especially those with the structure The chemical stability of the compounds is poor and they are prone to generate impurities, especially when they encounter alkaline conditions. This property of the compounds brings certain difficulties to drug development; some compounds with structural Although compounds in which the R group is an amino, carboxyl, or amide group have shown good activity in preliminary in vitro testing, their pharmacokinetic parameters are unsatisfactory, posing certain difficulties in drug development. In short, some compounds disclosed in patent application WO2017 / 114510A1 show certain drug development difficulties in comprehensive evaluation (for example, in terms of chemical stability and / or pharmacokinetic properties). Therefore, it is necessary to identify highly selective compounds with ERK kinase inhibitory activity that are more suitable for drug development through comprehensive evaluation. Summary of the Invention

[0004] After repeated experimental research, the inventors of the present invention finally found that the core structure Compounds in which R3 is a halogen, an unsubstituted alkyl, a halogenated alkyl, a deuterated alkyl, etc. have good chemical stability, good solubility and permeability, have ERK kinase inhibitory activity in both enzyme tests and cell tests, and have good pharmacokinetic parameters, and are particularly suitable for drug development.

[0005] Implementation Plan

[0006] In one aspect, the present invention provides novel ERK kinase inhibitors. Specifically, the present invention provides the following embodiments:

[0007] Embodiment 1. A compound of formula (I) or a stereoisomer, racemate, geometric isomer, tautomer, prodrug, hydrate, solvate or a pharmaceutically acceptable salt thereof,

[0008]

[0009] Where,

[0010] X1 selected from: CR 9a and N;

[0011] X2 selected from: CR 9b and N;

[0012] X3 selected from: CR 9cand N; and at most one of X1, X2 and X3 is N;

[0013] Y1 and Y2 are each independently selected from: CR9' and N;

[0014] R 9a 、R 9b and R 9c are each independently selected from: H, D, halogen, -OH, cyano, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted alkylcarbonyl, optionally substituted alkoxycarbonyl, optionally substituted cycloalkyl, amino, optionally substituted mono- or di-(alkyl)amino, and -CONR a R b ;

[0015] R9′ is selected from the group consisting of H, D, halogen, —OH, cyano, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted alkylcarbonyl, optionally substituted alkoxycarbonyl, optionally substituted cycloalkyl, amino, optionally substituted mono- or di-(alkyl)amino, and CONR a R b ;

[0016] R1 is selected from the group consisting of: H and D;

[0017] R2 is selected from the group consisting of optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl, or R2 and X1 together form an optionally substituted heterocyclyl;

[0018] R3 is selected from the group consisting of: halogen and optionally substituted alkyl;

[0019] R4 is selected from the group consisting of: H, D, optionally substituted alkyl, optionally substituted alkoxy, -CO(CR 10 R 11 ) m R 12 、-SO2(CR 10 R 11 ) m R 12 、-CONR 13 (CR 10 R 11 ) m R 12 、-COO(CR 10 R 11 ) m R 12 、-CR 13 R 13 '(CR 10 R 11 ) m R 12 and C1-8 Alkylcarbonyl-; wherein m is 0, 1, 2 or 3, and wherein

[0020] R 10 and R 11 are each independently selected from: H, D, halogen, optionally substituted alkyl and optionally substituted alkoxy; or R 10 With R 11 are joined to form optionally substituted cycloalkyl, cycloalkenyl, aryl, heteroaryl, and heterocyclyl; and

[0021] R 12 are each independently selected from: H, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl; and

[0022] R 13 and R 13 ' are each independently selected from: H and optionally substituted alkyl; or R 13 and R 13 'Together with adjacent carbon atoms, they form optionally substituted cycloalkyl, cycloalkenyl, and heterocyclyl groups;

[0023] R5, R6, R7 and R8 are each independently selected from: -H, -D, halogen, -OH, amino, cyano, optionally substituted alkyl, optionally substituted alkoxy, -(CH2) 0-3 CONR a R b 、-(CH2) 0-3 COOH, optionally substituted cycloalkyl and optionally substituted heterocyclyl; or any two of R5, R6, R7 and R8 together with adjacent carbons form optionally substituted cycloalkyl, cycloalkenyl, aryl, heteroaryl and heterocyclyl; and

[0024] R a and R b are each independently selected from: H, D, and optionally substituted alkyl;

[0025] wherein the optional substituents are independently selected from: deuterium (D), halogen, -OH, mercapto, cyano, -CD3, -C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, 3-8 membered cycloalkyl, aryl, 3-8 membered heterocyclyl, heteroaryl, aryl-C1-C6 alkyl-, heteroaryl-C1-C6 alkyl-, C1-C6 haloalkyl-, -OC1-C6 alkyl, -OC2-C6 alkenyl, -OC1-C6 alkylphenyl, -C1-C6 alkyl-OH, -C1-C 6 alkyl-SH, -C1-C6 alkyl-O-C1-C6 alkyl, -OC1-C6 haloalkyl, -NH2, -C1-C6 alkyl-NH2, -N(C1-C6 alkyl)2, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)(C1-C6 alkylphenyl), -NH(C1-C6 alkylphenyl), nitro, -C(O)-OH, -C(O)OC1-C6 alkyl, -CONRiRii (wherein Ri and Rii are H, D and C 1-6alkyl), -NHC(O)(C1-C6 alkyl), -NHC(O)(phenyl), -N(C1-C6 alkyl)C(O)(C1-C6 alkyl), -N(C1-C6 alkyl)C(O)(phenyl), -C(O)C1-C6 alkyl, -C(O)-5-7 membered heteroaryl), -C(O)C1-C6 alkylphenyl, -C(O)C1-C6 haloalkyl, -OC(O)C1-C6 alkyl, -S(O)2-C1-C6 alkyl, -S( -C1-C6 alkyl), -S(O)2-phenyl, -S(O)2-C1-C6 haloalkyl, -S(O)2NH2, -S(O)2NH(C1-C6 alkyl), -S(O)2NH(phenyl), -NHS(O)2(C1-C6 alkyl), -NHS(O)2(phenyl) and -NHS(O)2(C1-C6 haloalkyl), wherein each of the alkyl, cycloalkyl, phenyl, aryl, heterocyclyl and heteroaryl groups is optionally substituted by one or more selected Further substituted with the following substituents: halogen, -OH, -NH2, cycloalkyl, 3-8 membered heterocyclyl, C1-C4 alkyl, C1-C4 haloalkyl-, -OC1-C4 alkyl, -C1-C4 alkyl-OH, -C1-C4 alkyl-O-C1-C4 alkyl, -OC1-C4 haloalkyl, cyano, nitro, -C(O)-OH, -C(O)OC1-C6 alkyl, -CON(C1-C6 alkyl)2, -CONH(C1-C6 alkyl), -CONH2, -NHC(O)(C1-C6 alkyl), -NH(C1-C6 alkyl)C(O)(C1-C6 alkyl), -SO2(C1-C6 alkyl), -SO2(phenyl), -SO2(C1-C6 haloalkyl), -SO2NH2, -SO2NH(C1-C6 alkyl), -SO2NH(phenyl), -NHSO2(C1-C6 alkyl), -NHSO2(phenyl) and -NHSO2(C1-C6 haloalkyl),

[0026] with the proviso that the compound is not 2-(2-chloropyridin-3-yl)-1-(7-fluoro-2-(hydroxymethyl)-5-(2-(isopropylamino)pyrimidin-4-yl)indolin-1-yl)ethan-1-one.

[0027] Embodiment 2. The compound according to Embodiment 1 or its stereoisomer, racemate, geometric isomer, tautomer, prodrug, hydrate, solvate or pharmaceutically acceptable salt, characterized in that:

[0028] X1 selected from: CR 9a and N;

[0029] X2 selected from: CR 9b and N;

[0030] X3 selected from: CR 9c and N; and at most one of X1, X2 and X3 is N;

[0031] Y1 and Y2 are each independently selected from: CR9' and N;

[0032] R 9a 、R 9b and R 9c Each is independently selected from: H, D, halogen, -OH, cyano, optionally substituted C 1-3 Alkyl, optionally substituted C 1-3 Alkoxy, optionally substituted C 1-3 Alkylcarbonyl, optionally substituted C 1-3 Alkoxycarbonyl, optionally substituted C 3-8 Cycloalkyl, amino, optionally substituted mono- or di-(C 1-3 alkyl)amino and -CONR a R b ;

[0033] R9' is selected from the group consisting of: H, D, halogen, -OH, cyano, optionally substituted C 1-3 Alkyl, optionally substituted C 1-3 Alkoxy, optionally substituted C 1-3 Alkylcarbonyl, optionally substituted C 1-3 Alkoxycarbonyl, optionally substituted C 3-8 Cycloalkyl, amino, optionally substituted mono- or di-(C 1-3 alkyl)amino and CONR a R b ;

[0034] R1 is selected from the group consisting of: H and D;

[0035] R2 is selected from: optionally substituted C 1-8 Alkyl, optionally substituted C 3-8 cycloalkyl, optionally substituted 3-8 membered heterocyclyl, optionally substituted 6-12 membered aryl and optionally substituted 5-12 membered heteroaryl such as 5-7 membered heteroaryl, or R2 and X1 together form an optionally substituted 3-8 membered heterocyclyl;

[0036] R3 is selected from: halogen and C optionally substituted by one or more substituents independently selected from D and halogen 1-8 alkyl;

[0037] R4 is selected from the group consisting of: H, D, optionally substituted C 1-8 Alkyl, optionally substituted C 1-8 Alkoxy, -CO(CR 10 R 11 ) m R12 、-SO2(CR 10 R 11 ) m R 12 、-CONR 13 (CR 10 R 11 ) m R 12 、-COO(CR 10 R 11 ) m R 12 、-CR 13 R 13 '(CR 10 R 11 ) m R 12 and C 1-8 Alkylcarbonyl-; wherein m is 0, 1, 2 or 3, and wherein

[0038] R 10 and R 11 are each independently selected from: H, D, halogen, optionally substituted C 1-8 Alkyl and optionally substituted C 1-8 Alkoxy, or R 10 With R 11 are joined to form optionally substituted cycloalkyl, cycloalkenyl, aryl, heteroaryl, and heterocyclyl; and

[0039] R 12 are each independently selected from: H, optionally substituted C 1-3 Alkyl, optionally substituted C 3-8 cycloalkyl, optionally substituted 3-8 membered heterocyclyl, optionally substituted 6-12 membered aryl and optionally substituted 5-12 membered heteroaryl such as 5-7 membered heteroaryl; and

[0040] R 13 and R 13 ' are each independently selected from: H and optionally substituted C 1-3 Alkyl; or R 13 and R 13 Together with the adjacent carbons, it forms an optionally substituted C 3-8 Cycloalkyl, C 4-8 Cycloalkenyl and C 5-8 heterocyclic group;

[0041] R5, R6, R7 and R8 are each independently selected from: -H, -D, halogen, -OH, amino, cyano, optionally substituted C 1-3 Alkyl, optionally substituted C 1-3 Alkoxy, -(CH2) 0-3 CONRa R b 、-(CH2) 0-3 COOH, optionally substituted C 3-8 Cycloalkyl and optionally substituted 3-8 membered heterocyclyl; or any two of R5, R6, R7 and R8 together with adjacent carbon atoms form optionally substituted cycloalkyl, cycloalkenyl, aryl, heteroaryl and heterocyclyl; and

[0042] R a and R b are each independently selected from: H, D and optionally substituted C 1-3 alkyl.

[0043] Embodiment 3. The compound according to any one of Embodiments 1 to 2, or a stereoisomer, racemate, geometric isomer, tautomer, prodrug, hydrate, solvate, or pharmaceutically acceptable salt thereof, characterized in that the compound has a structure represented by Formula Ia, Ib, Ic, or Id:

[0044]

[0045] R 9a 、R 9b and R 9c Each independently selected from: H, D, C 1-3 Alkyl, C 1-3 Halogenated alkyl, C 1-3 Alkoxy, -OH, cyano, halogen, amino, mono- or di-(C 1-3 Alkyl)amino, C 1-3 Alkylcarbonyl, C 1-3 Alkoxycarbonyl and C 3-8 Cycloalkyl; preferably, R 9a 、R 9b and R 9c Each independently selected from: H, D and C 1-3 Alkyl; more preferably, R 9a 、R 9b and R 9c are each independently selected from: H and D; and

[0046] Other variables are as defined in Embodiment 1 or 2.

[0047] Embodiment 4. The compound according to any one of Embodiments 1 to 3, or a stereoisomer, racemate, geometric isomer, tautomer, prodrug, hydrate, solvate, or pharmaceutically acceptable salt thereof, characterized in that:

[0048] Y1 is CR9', Y2 is CR9', and R9' is selected from H, D, halogen and C 1-3Alkyl; more preferably, Y1 is CR9', Y2 is CR9', and wherein R9' is H, D, F or methyl;

[0049] or

[0050] Y1 is CR9', Y2 is N, and R9' is selected from H, D, halogen and C 1-3 Alkyl; more preferably, Y1 is CR9', Y2 is N, and wherein R9' is H, D, F or methyl;

[0051] or

[0052] Y1 is N, Y2 is CR9', and R9' is selected from H, D, halogen and C 1-3 Alkyl; more preferably, Y1 is N, Y2 is CR9', and wherein R9' is H, D, F or methyl;

[0053] or

[0054] Y1 is N, and Y2 is N.

[0055] Embodiment 5. The compound according to Embodiment 1 or its stereoisomer, racemate, geometric isomer, tautomer, prodrug, hydrate, solvate or pharmaceutically acceptable salt, characterized in that the compound has the structure shown in Formula Ie:

[0056]

[0057] wherein each variable is as defined in Embodiment 1.

[0058] Embodiment 6. The compound according to any one of Embodiments 1, 2 and 5, or its stereoisomer, racemate, geometric isomer, tautomer, prodrug, hydrate, solvate or pharmaceutically acceptable salt, characterized in that: X1 is selected from CR 9a and N, where R 9a Selected from H, D, halogen, -OH, cyano, C 1-3 Alkyl, C 1-3 Halogenated alkyl, C 1-3 Alkoxy, C 1-3 Alkylcarbonyl, C 1-3 Alkoxycarbonyl, C 3-8 Cycloalkyl, amino and mono- or di-(C 1-3 Preferably, X1 is selected from CR 9a and N, where R 9a Selected from H, D and C 1-3 Alkyl; more preferably, X1 is selected from CH, CD and N.

[0059] Embodiment 7. The compound according to any one of Embodiments 1 and 5, or a stereoisomer, racemate, geometric isomer, tautomer, prodrug, hydrate, solvate, or pharmaceutically acceptable salt thereof, characterized in that:

[0060] R2 is selected from: optionally substituted C 1-6 Alkyl, optionally substituted C 3-8 Cycloalkyl, optionally substituted 3-8 membered heterocyclyl and optionally substituted 5-12 membered heteroaryl such as 5-7 membered heteroaryl, wherein the optional substituents are one or more independently selected from D, halogen, hydroxy, -CD3, C 1-6 Alkyl and hydroxy C 1-6 The substituents of the alkyl group are preferably one or more substituents independently selected from D, halogen, hydroxy, -CD3, -CH3 and -CH2OH;

[0061] or

[0062] R2 is selected from: C optionally substituted by one or more hydroxyl groups 1-6 Alkyl, C optionally substituted by one or more hydroxyl groups 3-8 Cycloalkyl, 3-8 membered heterocyclic group and optionally one or more -CD3, C 1-6 Alkyl and hydroxy C 1-6 Alkyl-substituted 5-12 membered heteroaryl such as 5-7 membered heteroaryl;

[0063] or

[0064] R2 is selected from: C 1-4 alkyl, They are optionally replaced by one or more independently selected from D, halogen, hydroxy, C 1-4 Alkyl, -CD3 and hydroxyl C 1-4 Alkyl is substituted with one or more substituents independently selected from D, halogen, hydroxy, -CH3, -CD3 and -CH2OH;

[0065] or

[0066] R2 is selected from: isopropyl,

[0067] or

[0068] R2 is selected from

[0069] or

[0070] R2 is selected from

[0071] Embodiment 8. A compound according to any one of Embodiments 1 to 7, or a stereoisomer, racemate, geometric isomer, tautomer, prodrug, hydrate, solvate, or pharmaceutically acceptable salt thereof, characterized in that: R3 is selected from halogen and C optionally substituted with one or more substituents independently selected from D or halogen 1-6 Alkyl; or, R3 is selected from halogen and C optionally substituted by one or more substituents independently selected from D or halogen 1-3 Alkyl; or, R3 is selected from halogen and C 1-6 Alkyl; or R3 is selected from fluorine, chlorine, bromine, iodine, -CH3, -CH2CH3, -CH(CH3)2, -CH2CH2CH3, -CF3, -CHF2, CF3CH2- and CD3-; or, R3 is selected from fluorine, chlorine and -CH3; or, R3 is fluorine.

[0072] Embodiment 9. The compound of any one of Embodiments 1 to 8, or a stereoisomer, racemate, geometric isomer, tautomer, prodrug, hydrate, solvate, or pharmaceutically acceptable salt thereof, characterized in that:

[0073] R4 is selected from -CO(CR 10 R 11 ) m R 12 and-CR 13 R 13 '(CR 10 R 11 ) m R 12 ; where m is 0, 1, 2 or 3, and where

[0074] R 10 and R 11 are each independently selected from H, D and C optionally substituted by hydroxy 1-4 alkyl;

[0075] R 12 Each is independently selected from: optionally substituted 6-12 membered aryl and optionally substituted 5-12 membered heteroaryl such as 5-7 membered heteroaryl;

[0076] R 13 and R 13 ' are each independently selected from: H, C 1-3 Alkyl and C 1-3 Halogenated alkyl.

[0077] Embodiment 10. The compound according to Embodiment 9 or its stereoisomer, racemate, geometric isomer, tautomer, prodrug, hydrate, solvate or pharmaceutically acceptable salt, characterized in that: R 12Each is independently selected from optionally substituted 6-12 membered aryl and optionally substituted 5-12 membered heteroaryl such as 5-7 membered heteroaryl, wherein the optional substituent is one or more substituents independently selected from the following: D, halogen, C 1-4 Alkyl, cyano and C 3-8 Heterocyclyl-(CH2) 0-4 -(e.g. morpholinyl such as morpholino, piperazinyl, tetrahydropyranyl such as tetrahydropyran-4-yl, morpholinylmethyl such as morpholinomethyl or piperazinylmethyl);

[0078] Or, R 12 Selected from: optionally substituted phenyl and optionally substituted pyridinyl such as pyridin-3-yl, wherein the optional substituents are substituted by one or more substituents independently selected from the following substituents: D, halogen, C 1-4 Alkyl (such as methyl or ethyl), cyano and C 3-8 Heterocyclyl-(CH2) 0-4 -(e.g. morpholinyl such as morpholino, piperazinyl, tetrahydropyranyl such as tetrahydropyran-4-yl, morpholinylmethyl such as morpholinomethyl or piperazinylmethyl);

[0079] Or, R 12 Selected from: wherein Rc is selected from halogen such as fluorine or chlorine, C 1-4 Alkyl groups such as methyl, where R d Selected from H, C 1-4 Alkyl groups such as methyl or ethyl and where R e is selected from halogens such as fluorine and chlorine, and p is 1 or 2; and where R f Selected from

[0080] Embodiment 11. The compound according to any one of Embodiments 1 to 8, or a stereoisomer, racemate, geometric isomer, tautomer, prodrug, hydrate, solvate, or pharmaceutically acceptable salt thereof, characterized in that: R4 is selected from -CO(CR 10 R 11 ) m R 12 , where m is 0, 1, 2, or 3, and where

[0081] R 10 and R 11 are each independently selected from H; and

[0082] R 12 Selected from

[0083]

[0084] Embodiment 12. The compound of any one of Embodiments 1 to 8, or a stereoisomer, racemate, geometric isomer, tautomer, prodrug, hydrate, solvate, or pharmaceutically acceptable salt thereof, characterized in that: R4 is selected from -CO(CR 10 R 11 ) m R 12 , where m is 0, 1, 2 or 3, and where R 10 and R 11 are each independently selected from H; R 12 is selected from 2-cyanophenyl, 5-chloro-2-fluorophenyl, 2-chloro-3-fluorophenyl, 2-chloro-4-fluorophenyl, 2-chloro-5-fluorophenyl, 2,5-difluorophenyl, 3-chloropyridin-2-yl, 6-chloropyridin-2-yl, 3-chloropyridin-4-yl or 4-chloropyridin-3-yl.

[0085] Embodiment 13. The compound of any one of Embodiments 1 to 12, or a stereoisomer, racemate, geometric isomer, tautomer, prodrug, hydrate, solvate, or pharmaceutically acceptable salt thereof, characterized in that:

[0086] R5, R6, R7 and R8 are each independently selected from: -H, -D, halogen, -OH, amino, cyano, optionally substituted C 1-6 Alkyl, optionally substituted C 1-6 Alkoxy, -(CH2) 0-3 CONR a R b 、-(CH2) 0-3 COOH, optionally substituted C 3-8 Cycloalkyl and optionally substituted 3-8 membered heterocyclyl, wherein the optional substituents are one or more substituents independently selected from the group consisting of D, -OH, -OC1-C6 alkyl and NH2, and wherein R a and R b Each independently selected from H, D and C 1-3 alkyl;

[0087] Alternatively, R5, R6, R7 and R8 are each independently selected from H, C optionally substituted by hydroxy or -OC1-C6 alkyl. 1-6 alkyl;

[0088] Alternatively, R5 and R6 are each independently selected from H or C 1-6 and R7 and R8 are each independently selected from H and C optionally substituted by hydroxy or -OC1-C6 alkyl 1-6 alkyl;

[0089] Alternatively, R5, R6, R7 and R8 are each independently selected from -H, -CH3 and -CH2OH;

[0090] Alternatively, R5, R6 and R7 are H, and R8 is H, -CH3 or -CH2OH.

[0091] Embodiment 14. Compounds selected from Examples P1-P20, P23-P25, P28-51, P53-P64:

[0092]

[0093]

[0094]

[0095]

[0096]

[0097]

[0098]

[0099]

[0100]

[0101] or a pharmaceutically acceptable salt thereof.

[0102] On the other hand, the present invention also provides pharmaceutical compositions comprising the above novel compounds, uses of the above novel compounds, and methods of treatment using the above novel compounds:

[0103] Embodiment 15. A compound according to any one of Embodiments 1-14, or a pharmaceutically acceptable salt thereof, for use as a medicament.

[0104] Embodiment 16. A pharmaceutical composition comprising a compound according to any one of Embodiments 1-14 or a pharmaceutically acceptable salt thereof, and optionally comprising a pharmaceutically acceptable carrier.

[0105] Embodiment 17. Use of the compound according to any one of Embodiments 1 to 14 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing and / or treating diseases associated with ERK kinase or a product for use as an ERK kinase inhibitor.

[0106] Embodiment 18. A method for non-therapeutic inhibition of ERK kinase activity, comprising contacting an effective amount of a compound according to any one of Embodiments 1-14 or a pharmaceutically acceptable salt thereof with ERK kinase, thereby inhibiting ERK kinase.

[0107] On the other hand, the present invention also provides intermediates (e.g., intermediates 1-82 disclosed in the present invention, especially intermediate 25) and methods (e.g., Figure 1-4 The method shown in Figure 3 Method):

[0108] Embodiment 19. A compound having the following structure: tert-butyl (4-(7-fluoroindolin-5-yl)pyridin-2-yl)(1-methyl-1H-pyrazol-5-yl)carbamate or a stereoisomer, racemate, geometric isomer, tautomer, hydrate, solvate, or pharmaceutically acceptable salt thereof:

[0109]

[0110] Embodiment 20. A method for preparing a compound of formula (I) or a stereoisomer, racemate, geometric isomer, tautomer, prodrug, hydrate, solvate or pharmaceutically acceptable salt thereof as described in Embodiment 1, wherein the compound of formula (I) is a compound of formula C3:

[0111]

[0112] Among them, X1, R2, R3, R 10 、R 11 、R 12 and m is as defined in embodiment 1,

[0113] The method comprises the following steps:

[0114] (a) making a compound of formula C1

[0115]

[0116] With compound Carry out amide coupling reaction to generate a compound of formula C2,

[0117] and

[0118] (b) when the compound of formula C2 is Boc protected, deprotecting it to generate a compound of formula C3,

[0119]

[0120] Embodiment 21. The method of Embodiment 20, wherein the amide coupling reaction is carried out in the presence of a condensation reagent and a base in an inert solvent.

[0121] Embodiment 22. The method of embodiment 20, wherein the deprotection is performed in the presence of an acid in an inert solvent.

[0122] Embodiment 23. The method of embodiment 21 or 22, wherein the inert solvent is selected from: ethyl acetate, tetrahydrofuran, methyltetrahydrofuran, acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, dichloromethane, 1,2-dichloroethane, N-methyl-2-pyrrolidone, or a combination thereof.

[0123] Embodiment 24. The method of embodiment 21, wherein the condensation reagent is selected from the group consisting of: 1-hydroxybenzotriazole (HOBT), 1-hydroxy-7-azobenzotriazole (HOAT), benzotriazol-1-yl-oxytripyrrolidinium hexafluorophosphate (PyBOP), benzotriazole-1-tris(trimethylamino)-hexafluorophosphate (BOP), 1,1-carbonyldiimidazole (CDI), 1-propylphosphonic anhydride (T3P), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl), N,N-dicyclohexylcarbodiimide (DCC), acetic anhydride, acetyl chloride, oxalyl chloride, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate quaternary ammonium salt (HBTU).

[0124] Embodiment 25. The method of Embodiment 21, wherein the base is selected from one or more of: triethylamine, DIPEA, pyridine, 2,4-lutidine, NaOH, KOH, LiOH, Na2CO3, K2CO3, NaHCO3, Cs2CO3, Na3PO4, or K3PO4.

[0125] Embodiment 26. The method of Embodiment 20, wherein the amide coupling reaction is carried out at a temperature ranging from room temperature to reflux for 0.5 to 24 hours.

[0126] Embodiment 27. The method of Embodiment 22, wherein the acid is selected from one or more of hydrochloric acid, sulfuric acid, trifluoroacetic acid, acetic acid, formic acid, and phosphoric acid.

[0127] Embodiment 28. The method of embodiment 20, wherein the deprotection is at a temperature of -10°C to 80°C for 0.5 to 24 hours.

[0128] definition

[0129] The following terms and symbols used in this application have the meanings described below, unless the context indicates otherwise.

[0130] A hyphen ("-") that is not between two letters or symbols indicates the point of attachment of a substituent. For example, -O(C 1-3 Alkyl) refers to a C 1-3 However, when the attachment point of the substituent is obvious to those skilled in the art, for example, a halogen substituent, the "-" may be omitted.

[0131] When the group has a wavy line , the wavy line indicates the point of attachment of the group to the rest of the molecule.

[0132] The term "alkyl" as used herein refers to a linear or branched saturated monovalent hydrocarbon group having 1 to 8 carbon atoms, such as 1 to 6 carbon atoms, such as 1 to 4 carbon atoms, such as 1, 2 or 3 carbon atoms. 1-8 "Alkyl" means an alkyl group having 1 to 8 carbon atoms. Similarly, "C 1-4 "Alkyl" means an alkyl group having 1 to 4 carbon atoms; "C 1-3 The term "alkyl" refers to an alkyl group having 1 to 3 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl ("Me"), ethyl ("Et"), n-propyl ("n-Pr"), isopropyl ("i-Pr"), n-butyl ("n-Bu"), isobutyl ("i-Bu"), sec-butyl ("s-Bu"), tert-butyl ("t-Bu"), and the like. This definition applies regardless of whether the term "alkyl" is used alone or as part of another group, such as a haloalkyl group, an alkoxy group, and the like.

[0133] The term "alkenyl" as used herein refers to a linear or branched monovalent hydrocarbon group having 2 to 8 carbon atoms, such as 2 to 6 carbon atoms, such as 2, 3 or 4 carbon atoms, containing one or more, such as 1, 2 or 3, carbon-carbon double bonds (C=C). 2-6 "Alkenyl" means an alkenyl group having 2 to 6 carbon atoms and containing 1 or 2, preferably 1, carbon-carbon double bonds. Similarly, "C 2-3 "Alkenyl" means an alkenyl group having 2 to 3 carbon atoms and containing one carbon-carbon double bond. Examples of alkenyl groups include, but are not limited to, ethenyl, 2-propenyl, and 2-butenyl.

[0134] The term "alkynyl" as used herein refers to a linear or branched monovalent hydrocarbon group having 2 to 8 carbon atoms, such as 2 to 6 carbon atoms, such as 2 to 4 carbon atoms, containing one or more, such as 1, 2 or 3, carbon-carbon triple bonds (C≡C). 2-6"Alkynyl" means an alkynyl group having 2 to 6 carbon atoms and containing 1 or 2, preferably 1, carbon-carbon triple bonds. Similarly, "C 2-3 "Alkynyl" refers to an alkynyl group having 2 to 3 carbon atoms and containing one carbon-carbon triple bond. Examples of alkynyl groups include, but are not limited to, ethynyl, 2-propynyl, and 2-butynyl.

[0135] The term "alkoxy" as used herein refers to the group -O-alkyl, wherein alkyl is as defined above. For example, "C 1-8 "Alkoxy" means -OC 1-8 Alkyl, i.e., alkoxy having 1 to 8 carbon atoms. Similarly, "C 1-3 "Alkoxy" means -OC 1-3 Alkyl, i.e., an alkoxy group having 1 to 3 carbon atoms. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentoxy, and hexoxy. This definition applies regardless of whether the term "alkoxy" is used alone or as part of another group.

[0136] As used herein, the term "halo" or "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br) and iodine (I), preferably fluorine and chlorine, most preferably fluorine.

[0137] The term "haloalkyl" as used herein refers to an alkyl group as defined herein in which one or more hydrogen atoms, such as 1, 2, 3, 4 or 5 hydrogen atoms, are replaced by halogen, and when more than one hydrogen atom is replaced by a halogen atom, the halogen atoms may be the same or different from one another. Examples of haloalkyl groups include, but are not limited to, -CF3, -CHF2, -CH2CF3, and the like.

[0138] As used herein, the term "hydroxyl" refers to the group -OH.

[0139] As used herein, the term "mercapto" refers to the group -SH.

[0140] As used herein, the term "cyano" refers to the group -CN.

[0141] As used herein, the term "carboxyl" refers to the group -C(O)-OH, which may also be represented as -COOH.

[0142] As used herein, the term "carbonyl" refers to the group -C(O)-, which may also be represented as -CO-.

[0143] As used herein, the term "hydrogen" refers to the group -H.

[0144] As used herein, the symbol "D" refers to deuterium.

[0145] As used herein, the term "amino" refers to the group -NH2.

[0146] The term "alkylamino" or "monoalkylamino" as used herein refers to the group alkyl-NH-, wherein alkyl is as defined herein.

[0147] The term "dialkylamino" as used herein refers to the group (alkyl)2-N-, wherein alkyl is as defined herein.

[0148] The term "alkylcarbonyl" as used herein refers to an alkyl group attached to another group through a carbonyl group, ie, alkyl-C(O)-, wherein alkyl is as defined herein.

[0149] The term "alkoxycarbonyl" as used herein refers to an alkoxy group attached to another group through a carbonyl group, ie, alkoxy-C(O)-, wherein alkoxy is as defined herein.

[0150] As used herein, the term "oxo" refers to the group =0.

[0151] As used herein, the term "nitro" refers to the group -NO2.

[0152] The term "cycloalkyl" as used herein refers to a saturated monovalent monocyclic or bicyclic hydrocarbon group having 3 to 12 ring carbon atoms, such as 3 to 8 ring carbon atoms, such as 3 to 6 ring carbon atoms. 3-8 "Cycloalkyl" means a cycloalkyl group having 3 to 8 ring carbon atoms. Similarly, "C 3-6 "Cycloalkyl" means a cycloalkyl radical having 3 to 6 ring carbon atoms. Examples of cycloalkyl radicals include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[0153] The term "cycloalkenyl" as used herein refers to a cycloalkyl group as defined herein containing one or more double bonds, for example 1, 2, 3 or 4 double bonds, wherein the ring of the cycloalkenyl group is non-aromatic. For example, "C 3-8 "Cycloalkenyl" means a cycloalkenyl group having 3 to 8 ring carbon atoms. Similarly, "C 3-6 "Cycloalkenyl" means a cycloalkenyl group having 3 to 6 ring carbon atoms. Examples of cycloalkenyl groups include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl.

[0154] As used herein, the term "heterocyclyl" or "heterocyclic" or "heterocycle" refers to a monocyclic, bicyclic or tricyclic, saturated and partially unsaturated non-aromatic ring having 3-20 ring atoms, for example, 3-12 ring atoms, for example, 3-8 ring atoms, for example, 3-6 ring atoms, which contains at least one carbon atom in addition to 1-4, for example, 1-3, for example, 1 or 2, for example, 1 heteroatom selected from O, S and N. In one example, the "heterocyclyl" or "heterocyclic" or "heterocycle" refers to a monocyclic ring having 3 to 8 ring atoms, for example, 3, 4, 5 or 6 ring atoms, which contains at least one carbon atom in addition to 1-4, for example, 1-3, for example, 1 or 2, for example, 1 heteroatom selected from O, S and N. In one example, the "heterocyclyl" or "heterocyclic" or "heterocycle" contains 0, 1, 2 or 3 double bonds. Any nitrogen or sulfur heteroatom may optionally be oxidized (e.g., NO, SO, SO2), and any nitrogen heteroatom may optionally be quaternized (e.g., [NR4] + Cl - 、[NR4] + OH - A heterocyclic group having 3 to 8 ring atoms is also referred to as a 3-8 membered heterocyclic group. Heterocyclic groups having other numbers of carbon atoms may also be briefly expressed similarly. Examples of heterocyclic groups include, but are not limited to, oxiranyl, aziridinyl, thiirane, azetidinyl, oxetanyl, thietanyl, 1,2-dithietanyl, 1,3-dithietanyl, pyrrolidinyl (pyrrolidin-1-yl, pyrrolidin-2-yl, pyrrolidin-3-yl), dihydro-1H-pyrrolyl, dihydrofuranyl, tetrahydrofuranyl (e.g., tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrafluorofuran-4-yl), dihydrothiophenyl, tetrahydrothiophenyl, imidazolidinyl, piperidinyl, piperazinyl (e.g., piperazin-1-yl, piperazin-2-yl, piperazin-3-yl, piperazin-4-yl), isoquinolinyl, tetrahydroisoquinolinyl, morpholinyl ( For example, morpholino (i.e., morpholin-1-yl), morpholin-2-yl, morpholin-3-yl), thiomorpholinyl, 1,1-dioxo-thiomorpholinyl, dihydropyranyl, tetrahydropyranyl (e.g., tetrahydropyran-2-yl, tetrahydropyran-3-yl, tetrahydropyran-4-yl), hexahydrothiopyranyl, hexahydropyrimidinyl, oxazinanyl, thiazinanyl, thioxanyl, homopiperazinyl, homopiperidinyl, azepanyl, oxepanyl, thiepanyl, oxazepanyl, Oxazepanyl, diazepanyl, 1,4-diazepanyl, diazepanyl thiazolinone thiazepanyl, tetrahydrothiopyranyl, Oxazolidinyl, thiazolidinyl, isothiazolidinyl, 1,1-dioxoisothiazolidinone, oxazolidinone, imidazolidinone, 4,5,6,7-tetrahydro[2H]indazolyl, tetrahydrobenzimidazolyl, 4,5,6,7-tetrahydrobenzo[d]imidazolyl, 1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridinyl, thiazinyl, azine group, thiadiazine group, diazine, dithiazine, di Azine group, Thiazinyl, thiatriazinyl, triazinyl, dithiadiazinyl, imidazolinyl, dihydropyrimidinyl, tetrahydropyrimidinyl, 1-pyrrolinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, thiopyranyl, 2H-pyranyl, 4H-pyranyl, dihydropyrimidin ... alkyl, 1,3-dioxolanyl, pyrazolyl, pyrazolidinyl, dithianyl, dithiolanyl, pyrimidinyl, pyrimidinedione, pyrimidine-2,4-dione, piperazinyl, piperazindione, pyrazolidinyl imidazolinyl, 3-azabicyclo[3.1.0]hexanyl, 3,6-diazabicyclo[3.1.1]heptanyl, 6-azabicyclo[3.1.1]heptanyl, 3-azabicyclo[3.1.1]heptanyl, 3-azabicyclo[4.1.0]heptanyl, azabicyclo[2.2.2]hexanyl, 2-azabicyclo[3.1.0]hexanyl, cyclo[3.2.1]octyl, 8-azabicyclo[3.2.1]octyl, 2-azabicyclo[2.2.2]octyl, 8-azabicyclo[2.2.2]octyl, 7-oxabicyclo[2.2.1]heptane, azaspiro[3.5]nonyl, azaspiro[2.5]octyl, azaspiro[4.5]decyl, 1-azaspiro[4.5]decan-2-onyl, azaspiro[5.5]undecyl, tetrahydroindolyl, octahydroindolyl, tetrahydroisoindolyl, tetrahydroindazolyl, 1,1-dioxohexahydrothiopyranyl. Examples of 5-membered heterocyclic groups containing a sulfur or oxygen atom and 1 to 3 nitrogen atoms are: thiazolyl, including thiazol-2-yl and thiazol-2-yl N-oxide, thiadiazolyl, including 1,3,4-thiadiazol-5-yl and 1,2,4-thiadiazol-5-yl, Azolyl, e.g. oxazol-2-yl, and Oxazolyl, such as 1,3,4- oxadiazole-5-yl and 1,2,4- Examples of 5-membered heterocyclic groups containing 2 to 4 nitrogen atoms include: imidazolyl, such as imidazol-2-yl; triazolyl, such as 1,3,4-triazol-5-yl; 1,2,3-triazol-5-yl, 1,2,4-triazol-5-yl, and tetrazolyl, such as 1H-tetrazolyl. Examples of benzo-fused 5-membered heterocyclic groups are benzo Examples of 6-membered heterocyclic groups include oxazol-2-yl, benzothiazol-2-yl, and benzimidazol-2-yl. Exemplary 6-membered heterocyclic groups contain 1 to 3 nitrogen atoms and optionally contain sulfur or oxygen atoms, for example: pyridyl, such as pyridin-2-yl, pyridin-3-yl, and pyridin-4-yl; pyrimidinyl, such as pyrimidin-2-yl and pyrimidin-4-yl; triazinyl, such as 1,3,4-triazin-2-yl and 1,3,5-triazin-4-yl; pyridazinyl, particularly pyridazin-3-yl, and pyrazinyl. Pyridine N-oxides and pyridazine N-oxides, as well as pyridyl, pyrimidin-2-yl, pyrimidin-4-yl, pyridazinyl, and 1,3,4-triazin-2-yl are further examples of heterocyclic groups.

[0155] The term "hydroxyalkyl" as used herein refers to an alkyl group substituted with a hydroxy group, i.e., -alkyl-OH, wherein alkyl is as defined herein. Examples of such groups include, but are not limited to, hydroxymethyl, hydroxyethyl (e.g., 2-hydroxyethyl, 1-hydroxyethyl), hydroxypropyl (e.g., 1-hydroxypropyl-2-yl, 1-hydroxypropyl-3-yl, 1-hydroxypropyl-1-yl, etc.), hydroxybutyl (e.g., 4-hydroxybutyl-2-yl, etc.).

[0156] The term "aryl" as used herein refers to a carbocyclic hydrocarbon group consisting of one ring or a plurality of fused rings having 6 to 14 ring carbon atoms, for example, 6 to 12 ring carbon atoms, for example, 6 to 10 ring carbon atoms, wherein at least one ring is an aromatic ring. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl, and indenyl, with phenyl and naphthyl being preferred.

[0157] As used herein, the term "heteroaryl" refers to:

[0158] a monocyclic aromatic hydrocarbon group having 5, 6 or 7 ring atoms, for example having 6 ring atoms, which contains one or more, for example 1, 2 or 3, for example 1 or 2, ring heteroatoms independently selected from N, O and S (for example N) in the ring, the remaining ring atoms being carbon atoms; and

[0159] A bicyclic aromatic hydrocarbon group having 8 to 12 ring atoms, for example 9 or 10 ring atoms, which contains one or more, for example 1, 2, 3 or 4, for example 1 or 2, ring heteroatoms independently selected from N, O and S (for example N) in the ring, the remaining ring atoms being carbon atoms, wherein at least one ring is aromatic.

[0160] When the total number of S and O atoms in the heteroaryl group exceeds 1, these S and O heteroatoms are not adjacent to each other.

[0161] Heteroaryl groups also include those in which the N ring heteroatom is in the form of an N-oxide, for example N-oxidopyrimidinyl.

[0162] In some embodiments, the ring heteroatom in the above-mentioned heteroaryl group is an N atom, and such heteroaryl groups are referred to as "nitrogen-containing heteroaryl groups". Nitrogen-containing heteroaryl groups also include those heteroaryl groups in which the N ring heteroatom is in the form of an N-oxide, such as N-oxypyridyl. For example, the nitrogen-containing heteroaryl group is a monocyclic heteroaryl group having 5 ring atoms, which contains 1 or 2 N heteroatoms in the ring, and the remaining ring atoms are carbon atoms; for another example, the nitrogen-containing heteroaryl group is a monocyclic heteroaryl group having 6 ring atoms, which contains 1, 2, or 3 heteroatoms in the ring, and the remaining ring atoms are carbon atoms.

[0163] Examples of heteroaryl groups include, but are not limited to, pyridinyl (e.g., pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyridin-5-yl, pyridin-6-yl), pyridinyl N-oxide; pyrazinyl; pyrimidinyl; pyrazolyl (e.g., pyrazol-5-yl, pyrazol-1-yl, pyrazol-2-yl, pyrazol-3-yl, pyrazol-4-yl); imidazolyl; azole; iso oxazolyl; thiazolyl; isothiazolyl; thiadiazolyl; tetrazolyl; triazolyl; thienyl; furyl; pyranyl; pyrrolyl; pyridazinyl; benzo[d]thiazolyl; benzodioxolyl, for example benzo[d][1,3]dioxolyl; benzo Azolyl, such as benzo[d] oxazolyl; imidazopyridinyl, for example imidazo[1,2-a]pyridinyl; triazolopyridinyl, for example [1,2,4]triazolo[4,3-a]pyridinyl and [1,2,4]triazolo[1,5-a]pyridinyl; indazolyl; 2H-indazolyl; pyrrolopyrimidinyl, for example pyrrolo[3,4-d]pyrimidinyl, 7H-pyrrolo[2,3-d]pyrimidinyl; pyrazole pyrimidinyl, for example pyrazolo[1,5-a]pyrimidinyl; tetrazolopyridinyl, for example tetrazolopyridinyl; benzothienyl; benzofuranyl; benzimidazolinyl; indolyl; indolinyl; purinyl, for example 9H-purinyl and 7H-purinyl; quinolyl; isoquinolyl; 1,2,3,4-tetrahydroquinolyl and 5,6,7,8-tetrahydroisoquinolyl.

[0164] Examples of nitrogen-containing heteroaryl groups include, but are not limited to, pyrrolyl, pyrazolyl, imidazolyl, pyridyl, pyrazinyl, pyrimidinyl, N-oxidized pyrimidinyl, pyridazinyl, pyrrolopyrimidinyl, such as pyrrolo[3,4-d]pyrimidinyl and 7H-pyrrolo[2,3-d]pyrimidinyl, purinyl, such as 9H-purinyl and 7H-purinyl, quinolyl, indolyl, and indazolyl.

[0165] As used herein, "aryl" or "aromatic" follows Hückel's rule, where the number of π electrons is equal to 4n+2, and n is zero or any positive integer up to 6.

[0166] As used herein, the terms "optional," "optionally," or "optionally" mean that the subsequently described substitution pattern, event, or circumstance may or may not occur, and that the description includes instances where the substitution pattern occurs as well as instances where the substitution pattern does not occur. For example, "optionally substituted alkyl" includes "unsubstituted alkyl" and "substituted alkyl" as defined herein. It will be understood by those skilled in the art that, for any group containing one or more substituents, the group does not include any substitution pattern that is sterically impractical, chemically incorrect, synthetically infeasible, and / or inherently unstable.

[0167] As used herein, the term "substituted" or "substituted by" means that one or more hydrogen atoms on a given atom or group are replaced by one or more substituents selected from a given group of substituents, provided that the normal valence of the given atom is not exceeded. When the substituent is oxo (i.e., =O), then two hydrogen atoms on a single atom are replaced by oxygen. Such combinations are permitted only if the combination of substituents and / or variables results in a chemically correct and stable compound. A chemically correct and stable compound means that the compound is stable enough to be isolated from a reaction mixture and the chemical structure of the compound can be determined, and then it can be formulated into a formulation that has at least practical utility. For example, in the absence of a clear listing of substituents, the term "substituted" or "substituted by" as used herein means that one or more hydrogen atoms on a given atom or group are independently replaced by one or more, for example, 1, 2, 3 or 4, substituents independently selected from: deuterium (D), halogen, -OH, mercapto, cyano, -CD3, -C1-C6 alkyl (preferably -C 1-3C2-C6 alkyl), C2-C6 alkenyl, C2-C6 alkynyl, cycloalkyl (preferably 3-8 membered cycloalkyl), aryl, heterocyclyl (preferably 3-8 membered heterocyclyl), heteroaryl, aryl-C1-C6 alkyl-, heteroaryl-C1-C6 alkyl-, C1-C6 haloalkyl-, -OC1-C6 alkyl (preferably -OC1-C3 alkyl), -OC2-C6 alkenyl, -OC1-C6 alkylphenyl, -C1-C6 alkyl-OH (preferably -C1-C4 alkyl-OH), -C1-C6 alkyl-SH, -C1-C6 alkyl-O-C1-C6 alkyl, -OC1-C 6 haloalkyl, -NH2, -C1-C6 alkyl-NH2 (preferably -C1-C3 alkyl-NH2), -N(C1-C6 alkyl)2 (preferably -N(C1-C3 alkyl)2), -NH(C1-C6 alkyl) (preferably -NH(C1-C3 alkyl)), -N(C1-C6 alkyl)(C1-C6 alkylphenyl), -NH(C1-C6 alkylphenyl), nitro, -C(O)-OH, -C(O)OC1-C6 alkyl (preferably -C(O)OC1-C3 alkyl), -CONRiRii (wherein Ri and Rii are H, D and C 1-6 Alkyl, preferably C 1-3C1-C6 alkyl), -NHC(O)(C1-C6 alkyl), -NHC(O)(phenyl), -N(C1-C6 alkyl)C(O)(C1-C6 alkyl), -N(C1-C6 alkyl)C(O)(phenyl), -C(O)C1-C6 alkyl, -C(O)heteroaryl (preferably -C(O)-5-7 membered heteroaryl), -C(O)C1-C6 alkylphenyl, -C(O)C1-C6 haloalkyl, -OC(O)C1-C6 alkyl (preferably -OC(O)C1-C3 alkyl) -C1-C6 alkyl), -S(O)2-C1-C6 alkyl, -S(O)2-phenyl, -S(O)2-C1-C6 haloalkyl, -S(O)2NH2, -S(O)2NH(C1-C6 alkyl), -S(O)2NH(phenyl), -NHS(O)2(C1-C6 alkyl), -NHS(O)2(phenyl) and -NHS(O)2(C1-C6 haloalkyl), wherein the alkyl, cycloalkyl, phenyl, aryl, heterocyclic and heteroaryl groups are Each of the following is optionally further substituted by one or more substituents selected from the group consisting of halogen, -OH, -NH2, cycloalkyl, 3-8 membered heterocyclyl, C1-C4 alkyl, C1-C4 haloalkyl-, -OC1-C4 alkyl, -C1-C4 alkyl-OH, -C1-C4 alkyl-O-C1-C4 alkyl, -OC1-C4 haloalkyl, cyano, nitro, -C(O)-OH, -C(O)OC1-C6 alkyl, -CON(C1-C6 alkyl)2, -CONH(C1 -C6 alkyl), -CONH2, -NHC(O)(C1-C6 alkyl), -NH(C1-C6 alkyl)C(O)(C1-C6 alkyl), -SO2(C1-C6 alkyl), -SO2(phenyl), -SO2(C1-C6 haloalkyl), -SO2NH2, -SO2NH(C1-C6 alkyl), -SO2NH(phenyl), -NHSO2(C1-C6 alkyl), -NHSO2(phenyl) and -NHSO2(C1-C6 haloalkyl). When an atom or group is substituted with a plurality of substituents, the substituents may be the same or different.

[0168] As used herein, the term "pharmaceutically acceptable" means non-toxic, biologically tolerable, and suitable for administration to an individual.

[0169] The term "pharmaceutically acceptable salt" as used herein refers to non-toxic, biologically tolerable acid addition salts or base addition salts of compounds of formula (I) suitable for administration to a subject, including but not limited to: acid addition salts of compounds of formula (I) formed with inorganic acids, such as hydrochlorides, hydrobromides, carbonates, bicarbonates, phosphates, sulfates, sulfites, nitrates, etc.; and acid addition salts of compounds of formula (I) formed with organic acids, such as formate, acetate, malate, maleate, fumarate, tartrate, succinate, citrate, lactate, methanesulfonate, p-toluenesulfonate, 2-hydroxyethanesulfonate, benzoate, salicylate, stearate and compounds of formula (HOOC-(CH2)) n "Pharmaceutically acceptable salts" also include base addition salts formed by compounds of formula (I) with acidic groups and pharmaceutically acceptable cations such as sodium, potassium, calcium, aluminum, lithium and ammonium.

[0170] Furthermore, if the compounds described herein are obtained in the form of acid addition salts, their free base forms can be obtained by basifying a solution of the acid addition salt. Conversely, if the product is in the form of a free base, its acid addition salt, particularly a pharmaceutically acceptable acid addition salt, can be obtained by dissolving the free base in a suitable solvent and treating the solution with an acid according to conventional procedures for preparing acid addition salts from basic compounds. Those skilled in the art can readily identify various synthetic methods for preparing non-toxic pharmaceutically acceptable acid addition salts without undue experimentation.

[0171] The compounds of the present invention may exist in the form of solvates. The term "solvate" refers to a solvent addition form comprising a stoichiometric or non-stoichiometric amount of a solvent. If the solvent is water, the solvate formed is a hydrate, and when the solvent is ethanol, the solvate formed is an ethanolate. Hydrates are formed by one or more molecules of water with one molecule of the substance, wherein the water retains its molecular form of HO. Such a combination can form one or more hydrates, such as hemihydrates, monohydrates, and dihydrates.

[0172] The term "prodrug" as used herein refers to an active or inactive compound that is chemically modified into a compound of the present invention by physiological actions in the body, such as hydrolysis, metabolism, etc., after administration to a subject. The suitability and techniques involved in preparing and using prodrugs are well known to those skilled in the art. Exemplary prodrugs include, for example, esters of free carboxylic acids and S-acyl derivatives of thiols and O-acyl derivatives of alcohols or phenols. Suitable prodrugs are generally pharmaceutically acceptable ester derivatives that can be converted into the parent carboxylic acid by solvolysis under physiological conditions, such as lower alkyl esters, cycloalkyl esters, lower alkenyl esters, benzyl esters, mono- or di-substituted lower alkyl esters, such as ω-(amino, mono- or di-lower alkylamino, carboxyl, lower alkoxycarbonyl)-lower alkyl esters, α-(lower alkanoyloxy, lower alkoxycarbonyl or di-lower alkylaminocarbonyl)-lower alkyl esters, such as pivaloyloxymethyl esters, which are conventionally used in the art.

[0173] It will be appreciated by those skilled in the art that some compounds of formula (I) may contain one or more chiral centers and therefore exist as two or more stereoisomers. Therefore, the compounds of the present invention may exist as single stereoisomers (e.g., enantiomers, diastereomers) and mixtures thereof in any proportion, such as racemates, and, where appropriate, as tautomers and geometric isomers.

[0174] As used herein, the term "stereoisomers" refers to compounds that have identical chemical constitution but differ in the arrangement of the atoms or groups in space. Stereoisomers include enantiomers, diastereomers, conformers, etc.

[0175] As used herein, the term "enantiomers" refers to two stereoisomers of a compound that are non-superimposable mirror images of one another.

[0176] As used herein, the term "diastereomer" refers to stereoisomers that have two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers have different physical properties, such as melting points, boiling points, spectral properties, or biological activities. Mixtures of diastereomers can be separated using high-resolution analytical methods such as electrophoresis and chromatography, such as HPLC.

[0177] Stereochemical definitions and conventions can be followed in SP Parker, ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994. Many organic compounds exist in optically active forms, that is, they have the ability to rotate the plane of plane-polarized light. When describing optically active compounds, the prefixes D and L or R and S are used to indicate the absolute configuration of the molecule about its chiral center. The prefixes d and l or (+) and (-) are used to indicate the sign of rotation of plane-polarized light by the compound, where (-) or l indicates that the compound is left-handed. Compounds with a prefix of (+) or d are right-handed. For a given chemical structure, these stereoisomers are identical except that they are mirror images of each other. A specific stereoisomer may also be referred to as an enantiomer, and a mixture of such isomers is often referred to as an enantiomeric mixture. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which can occur when there is no stereoselectivity or stereospecificity in a chemical reaction or process. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomers that are optically inactive.

[0178] The racemic mixture can be used as such or resolved into its individual isomers. Resolution can yield a stereochemically pure compound or a mixture enriched in one or more isomers. Methods for separating isomers are well known (see Allinger NL and Eliel EL, "Topics in Stereochemistry," Vol. 6, Wiley Interscience, 1971) and include physical methods such as chromatography using chiral adsorbents. Individual isomers can be prepared in chiral form from chiral precursors. Alternatively, the individual isomers can be chemically separated from the mixture by forming diastereomeric salts with chiral acids (e.g., individual enantiomers of 10-camphorsulfonic acid, camphoric acid, α-bromocamphoric acid, tartaric acid, diacetyltartaric acid, malic acid, pyrrolidone-5-carboxylic acid, etc.), fractionally crystallizing the salts, then liberating one or both of the resolved bases, and optionally repeating this process to obtain one or both isomers substantially free of the other isomer, i.e., the desired stereoisomer having an optical purity of, for example, at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 99.5% by weight. Alternatively, as is well known to those skilled in the art, the racemate can be covalently linked to a chiral compound (auxiliary) to obtain diastereomers, which can be separated by chromatography or fractional crystallization, followed by chemical removal of the chiral auxiliary to obtain the pure enantiomers.

[0179] The term "conformational isomer" as used herein refers to isomers formed by different spatial positions of atoms or groups in the molecules of covalently bonded compounds connected by single bonds, such as the semi-chair conformer and envelope conformer of cyclopentane.

[0180] As used herein, the term "tautomer" or "tautomeric form" refers to structural isomers of different energies that are interconvertible via a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) include interconversions via migration of a proton, such as keto-enol and imine-enamine isomerizations. Valence tautomers include interconversions via reorganization of some of the bonding electrons.

[0181] The term "geometric isomers" used herein refers to isomers caused by the inability of double bonds or single bonds of ring carbon atoms to rotate freely, also known as cis-trans isomers. Substituent groups located on the same side of the plane are cis isomers, and substituent groups located on opposite sides of the plane are trans isomers.

[0182] As used herein, the term "treating" refers to administering one or more pharmaceutical substances, particularly compounds of Formula (I) as described herein and / or pharmaceutically acceptable salts thereof, to a subject suffering from a disease or symptoms of the disease in order to cure, alleviate, relieve, alter, cure, ameliorate, improve, or affect the disease or symptoms of the disease. In some embodiments, the disease is cancer.

[0183] As used herein, the term "prevention" refers to administering one or more pharmaceutical substances, particularly compounds of formula (I) and / or pharmaceutically acceptable salts thereof, to an individual with a predisposition to the disease to prevent the individual from developing the disease. In some embodiments, the disease is a disease associated with ERK kinases, such as ERK1 and / or ERK2 kinases. In further embodiments, the disease is a disease associated with high expression or high activity of ERK kinases, such as ERK1 and / or ERK2 kinases. In further embodiments, the disease is cancer or a tumor.

[0184] As used herein, the terms "cancer," "carcinoma," and "tumor" refer to the physiological condition in mammals that is typically characterized by unregulated cell growth. Examples of such cancers include blastomas, gliomas, sarcomas, seminoma, glioblastomas, melanomas, leukemias, and myeloid or lymphoid malignancies. More specific examples of such cancers include squamous cell carcinomas (e.g., epithelial squamous cell carcinomas) and lung cancers, including small cell lung cancer, non-small cell lung cancer (NSCLC), lung adenocarcinomas, and lung squamous cell carcinomas. Additional cancers include skin cancer, keratoacanthoma, follicular carcinoma, hairy cell leukemia, oral vestibule cancer, pharyngeal cancer, lip cancer, tongue cancer, mouth cancer, salivary gland cancer, esophageal cancer, laryngeal cancer, hepatocellular carcinoma, stomach cancer, gastrointestinal cancer, small intestine cancer, large intestine cancer, pancreatic cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, genitourinary system cancer, biliary tract cancer, gallbladder adenocarcinoma, thyroid cancer, papillary carcinoma, endometrial cancer, uterine cancer, salivary gland cancer, kidney cancer, prostate cancer, testicular cancer, vulvar cancer, peritoneal cancer, anal cancer, penile cancer, bone cancer, multiple myeloma, B-cell lymphoma, central nervous system cancer, brain cancer, head and neck cancer, and Hodgkin lymphoma. Examples also include myeloproliferative disorders such as polycythemia vera, essential thrombocythemia, myelofibrosis such as primary myelofibrosis, acute myeloid leukemia, and chronic myeloid leukemia (CML).

[0185] When referring to a chemical reaction, the terms "treating," "contacting," and "reacting" mean adding or mixing two or more reagents under appropriate conditions to produce the indicated and / or desired product. It should be understood that the reaction that produces the indicated and / or desired product may not necessarily result directly from the combination of the two reagents initially added, that is, there may be one or more intermediates generated in the mixture that ultimately lead to the formation of the indicated and / or desired product.

[0186] The term "effective amount" as used herein refers to an amount that is generally sufficient to produce a beneficial effect on an individual. The effective amount of a compound of the present invention can be determined by conventional methods (e.g., modeling, dose escalation studies, or clinical trials) in combination with conventional influencing factors (e.g., route of administration, pharmacokinetics of the compound, severity and course of the disease, individual medical history, individual health status, individual response to the drug, etc.).

[0187] The term "inhibit" refers to a decrease in the baseline activity of a biological activity or process. The term "inhibit ERK activity" refers to a decrease in ERK activity caused by a direct or indirect response to the presence of a compound of formula (I) or a pharmaceutically acceptable salt thereof as described herein, relative to the activity of ERK in the absence of a compound of formula (I) or a pharmaceutically acceptable salt thereof. The decrease in activity can be caused by a direct interaction between a compound of formula (I) or a pharmaceutically acceptable salt thereof as described herein and ERK, or by an interaction between a compound of formula (I) and / or a pharmaceutically acceptable salt thereof as described herein and one or more other factors that affect ERK activity.

[0188] As used herein, the term "subject" refers to both mammals and non-mammals. A mammal refers to any member of the class mammalia, including but not limited to humans; non-human primates such as chimpanzees and other apes and monkeys; farm animals such as cattle, horses, sheep, goats, and pigs; livestock such as rabbits, dogs, and cats; laboratory animals, including rodents such as rats, mice, and guinea pigs; and the like. Examples of non-mammals include, but are not limited to, birds. The term "subject" does not limit the subject to a particular age or sex. In some embodiments, the subject is a human.

[0189] In general, the term "about" is used herein to modify a stated numerical value by 20% above or below that value.

[0190] Technical and scientific terms used herein without specific definition have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0191] General synthetic method

[0192] The compound of formula (I) of the present invention or its stereoisomer, racemate, geometric isomer, tautomer, prodrug, hydrate, solvate or its pharmaceutically acceptable salt can be prepared by a variety of methods, including the methods given below, the methods given in the examples or similar methods. A suitable general synthesis scheme is described below. Suitable reaction conditions for each reaction step are known to those skilled in the art. The raw materials can be purchased or can be prepared by the methods below, methods similar to the methods given below or methods known in the art. Each variable in the general formula has the same meaning as above, unless otherwise specified.

[0193] The general synthesis method of the compound of the present invention is shown in the attached Figure 1-4 .

[0194] Figure 1 A general synthetic scheme A for synthesizing compounds of the present invention is shown.

[0195] Figure 2 General Synthetic Scheme B for synthesizing compounds of the present invention is shown.

[0196] Figure 3 A general synthetic scheme C for synthesizing compounds of the present invention is shown.

[0197] Figure 4 General Synthetic Scheme D is shown for the synthesis of compounds of the present invention.

[0198] In these schemes, it is fully understood that: when necessary, protecting groups for sensitive or reactive groups (such as amino, hydroxyl and carboxyl groups) are used according to general principles or chemistry to prevent them from undesirable chemical reactions. Protecting groups are treated according to standard methods of organic synthesis (TW Greene and PGM Wuts, "Protective Groups in Organic Synthesis", 5th edition, Wiley, New York 2014). These groups are removed using methods well known to those skilled in the art at a convenient stage in the synthesis of the compound. The selection of methods, reaction conditions and the order in which they are carried out should be consistent with the preparation of the compound of formula (I).

[0199] The example of amino protecting group comprises carbamate, amide, alkyl and aryl, and imines, and many N-heteroatom derivatives, and it can be removed to regenerate required amine group.Specific amino protecting group is Pmb (p-methoxybenzyl), Boc (tert-butyloxycarbonyl), Fmoc (9-fluorenylmethoxycarbonyl) and Cbz (benzyloxycarbonyl).The other example of these groups sees TW Greene and PGM Wuts, " Protecting Groups in Organic Synthesis ", the 3rd edition, John Wiley & Sons, Inc., in 1999.

[0200] Examples of hydroxy protecting groups include tetrahydropyranyloxy, benzoyl, acetoxy, carbamoyloxy, benzyl, and silyl ether (e.g., TBS, TBDPS) groups. Additional examples of these groups are found in TW Greene and PGM Wuts, "Protecting Groups in Organic Synthesis, 3rd Edition, John Wiley & Sons, Inc., 1999.

[0201] Examples of carboxyl protecting groups include ester groups and heterocyclic groups. Ester derivatives of carboxylic acid groups can be used to block or protect the carboxylic acid group while reactions are carried out on other functional groups of the compound. Examples of such ester groups include: substituted arylalkyl groups, including substituted benzyl groups, such as 4-nitrobenzyl, 4-methoxybenzyl, 3,4-dimethoxybenzyl, 2,4-dimethoxybenzyl, 2,4,6-trimethoxybenzyl, 2,4,6-trimethylbenzyl, pentamethylbenzyl, 3,4-methylenedioxybenzyl; alkyl or substituted alkyl esters, such as methyl, ethyl, tert-butylallyl or tert-amyl, triphenylmethyl (trityl), 4-methoxytrityl, 4,4'-dimethoxytrityl, 4,4',4"-trimethoxytrityl, 2-phenylprop-2-yl; thioesters, such as tert-butylthioester; silyl esters, such as trimethylsilyl ester, tert-butyldimethylsilyl ester (TBSO), and the like. Additional examples of these groups can be found in TW Greene and PGM Wuts, "Protective Groups in Organic Synthesis”, 5th edition, Wiley, New York, 2014.

[0202] Those skilled in the art will recognize whether a stereocenter exists in a compound of formula (I). Thus, when a compound is desired as a single enantiomer or diastereomer, it can be obtained by stereoselective synthesis or by resolving the final product or any convenient intermediate. Resolving the final product, intermediate, or starting material can be performed by any suitable method known in the art. See, for example, E.L. Eliel, S.H. Wilen, and L.N. Mander, "Stereochemistry of Organic Compounds" (Wiley-Interscience, 1994).

[0203] exist Figure 3 In the method of embodiment 20, compound C2 is prepared by reacting compound C1 with a carboxylic acid This is achieved by an amide coupling reaction.

[0204] Preferably, the amide coupling reaction is carried out in an inert solvent. More preferably, the amide coupling reaction is carried out in the presence of a condensation reagent and a base in an inert solvent.

[0205] The inert solvent is preferably selected from the group consisting of ethyl acetate, tetrahydrofuran, methyltetrahydrofuran, acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, dichloromethane, 1,2-dichloroethane, N-methyl-2-pyrrolidone, or a combination thereof.

[0206] The condensation reagent is preferably selected from the group consisting of 1-hydroxybenzotriazole (HOBT), 1-hydroxy-7-azobenzotriazole (HOAT), benzotriazole-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP), benzotriazole-1-tris(trimethylamino)-hexafluorophosphate (BOP), 1,1-carbonyldiimidazole (CDI), 1-propylphosphonic anhydride (T3P), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl), N,N-dicyclohexylcarbodiimide (DCC), acetic anhydride, acetyl chloride, oxalyl chloride, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), and O-(benzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate quaternary ammonium salt (HBTU), more preferably T3P.

[0207] The base is preferably selected from one or more of triethylamine, DIPEA, pyridine, 2,4-lutidine, NaOH, KOH, LiOH, Na2CO3, K2CO3, NaHCO3, Cs2CO3, Na3PO4 or K3PO4; more preferably DIPEA.

[0208] Preferably, the amide coupling reaction is carried out at a temperature ranging from room temperature to reflux.

[0209] Preferably, the reaction time of the amide coupling reaction is 0.5 to 24 hours.

[0210] exist Figure 3 In the method of embodiment 20, when the compound of formula C2 is Boc-protected, compound C3 can be prepared by deprotecting compound C2.

[0211] Preferably, the deprotection is carried out in an inert solvent, which is preferably selected from the group consisting of ethyl acetate, tetrahydrofuran, methyltetrahydrofuran, acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, dichloromethane, 1,2-dichloroethane, N-methyl-2-pyrrolidone, or a combination thereof.

[0212] Preferably, the deprotection is carried out in the presence of an acid, which is preferably selected from one or more of hydrochloric acid, sulfuric acid, trifluoroacetic acid, acetic acid, formic acid, and phosphoric acid.

[0213] Preferably, the deprotection is carried out at a temperature of -10°C to 80°C.

[0214] Preferably, the deprotection is carried out for 0.5 to 24 hours.

[0215] Efficacy and administration

[0216] The compounds of the present invention can be used to treat diseases associated with ERK kinases such as ERK1 and / or ERK2 kinases, for example, diseases associated with high expression or high activity of ERK kinases such as ERK1 and / or ERK2 kinases, such as tumors and cancers. More specifically, the tumors and cancers are selected from, for example, blastomas, gliomas, sarcomas, seminoma, glioblastomas, melanomas, leukemias, and myeloid or lymphoid malignancies. More specific examples of such cancers include squamous cell carcinomas (e.g., epithelial squamous cell carcinomas) and lung cancers, including small cell lung cancer, non-small cell lung cancer (NSCLC), lung adenocarcinomas, and lung squamous cell carcinomas. Additional cancers include skin cancer, keratoacanthoma, follicular carcinoma, hairy cell leukemia, oral vestibule cancer, pharyngeal cancer, lip cancer, tongue cancer, mouth cancer, salivary gland cancer, esophageal cancer, laryngeal cancer, hepatocellular carcinoma, stomach cancer, gastrointestinal cancer, small intestine cancer, large intestine cancer, pancreatic cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, genitourinary system cancer, biliary tract cancer, gallbladder adenocarcinoma, thyroid cancer, papillary carcinoma, endometrial cancer, uterine cancer, salivary gland cancer, kidney cancer, prostate cancer, testicular cancer, vulvar cancer, peritoneal cancer, anal cancer, penile cancer, bone cancer, multiple myeloma, B-cell lymphoma, central nervous system cancer, brain cancer, head and neck cancer, and Hodgkin lymphoma. Examples also include myeloproliferative disorders such as polycythemia vera, essential thrombocythemia, myelofibrosis such as primary myelofibrosis, acute myeloid leukemia, and chronic myeloid leukemia (CML).

[0217] The compounds of the present invention may be administered to an individual in the form of a pharmaceutical composition, which may optionally contain one or more pharmaceutically acceptable excipients.

[0218] The compounds of the present invention can be administered by various known routes, including oral, rectal, intragastric, intracranial and parenteral administration, such as intravenous, intramuscular, intranasal, intradermal, subcutaneous, and similar routes of administration. Particularly preferred are oral, intranasal and parenteral administration. Depending on the route of administration, different pharmaceutical formulations are required, and some of these routes of administration may require the application of a protective coating to the pharmaceutical formulation to prevent degradation of the compounds of the present invention, such as in the digestive tract.

[0219] The compound of the present invention can be formulated into syrup, infusion or injection solution, spray, tablet, capsule, lozenge, liposome, suppository and the like.

[0220] The particularly preferred pharmaceutical form for administering the compound of the present invention is a form suitable for injection use, including aseptic aqueous solutions or dispersions and the sterile powder for the immediate preparation of sterile injection solutions or dispersions. In all cases, final solution or dispersion form must be aseptic and are fluid. Typically, this type of solution or dispersion will comprise a solvent or dispersion medium containing, for example, a water-buffered aqueous solution such as a biocompatible buffer, ethanol, a polyol such as glycerol, propylene glycol, polyethylene glycol, a mixture thereof, a surfactant or a vegetable oil. The compound of the present invention can also be formulated into liposomes, particularly liposomes for parenteral administration. Liposomes provide the advantage of increased half-life (if compared with free drugs) in circulation and the more uniform release of the extended duration of the encapsulated medicine.

[0221] Sterilization of infusions and injections can be achieved by techniques recognized in the art, including but not limited to the addition of preservatives such as antibacterial or antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid or thimerosal. In addition, isotonic agents, such as sugars or salts, particularly sodium chloride, can also be incorporated into infusions and injections.

[0222] Sterile injectable solutions containing one or more compounds of the present invention are prepared by incorporating the desired amount of each compound into a suitable solvent containing the ingredients listed above, as appropriate, followed by sterilization. To obtain a sterile powder, the solution is vacuum dried or freeze-dried as needed. Preferred diluents for the present invention are water, a physiologically acceptable buffer, a physiologically acceptable buffered saline solution, or a saline solution. Preferred carriers are cocoa butter and vitebesole.

[0223] Excipients that can be used with the various pharmaceutical forms of the compounds of the present invention can be selected from the following non-limiting list: a) binders, such as lactose, mannitol, crystalline sorbitol, hydrogen phosphate, sugars, microcrystalline cellulose, carboxymethyl cellulose, hydroxyethyl cellulose, polyvinyl pyrrolidone, and the like;

[0224] b) lubricants, such as magnesium stearate, talc, calcium stearate, zinc stearate, stearic acid, hydrogenated vegetable oils, leucine, glycerides, and sodium stearyl fumarate;

[0225] c) disintegrants, such as starch, cross-linked carboxymethyl cellulose, sodium methyl cellulose, agar, bentonite, alginic acid, carboxymethyl cellulose, polyvinyl pyrrolidone, etc.

[0226] In one embodiment, the formulation is for oral administration and comprises one or more or all of the following ingredients: pregelatinized starch, talc, polyvinylpyrrolidone K30, croscarmellose sodium, sodium stearyl fumarate, gelatin, titanium dioxide, sorbitol, monosodium citrate, xanthan gum, titanium dioxide, flavoring, sodium benzoate, and sodium saccharin.

[0227] In one embodiment, the compounds of the invention are administered intranasally, which can be administered with a dry powder inhaler or from a nasal cannula using a suitable propellant such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, a hydrofluoroalkane such as 1,1,1,2-tetrafluoroethane (HFA 134A) or a combination thereof. TM ) or 1,1,1,2,3,3,3-heptafluoropropane (HFA 227EA TM ), carbon dioxide, or other suitable gas in a pressurized container, pump, sprayer or atomizer. The pressurized container, pump, sprayer or atomizer may contain a solution or suspension of the compound of the invention, such as a solution or suspension using ethanol and a propellant as a solvent, and may further contain a lubricant, such as sorbitan trioleate.

[0228] Typical dosage ranges for the compounds of the present invention are 0.001-1000 mg active ingredient / kg body weight / day. These doses may be administered once daily or in divided doses. The appropriate dosage is determined by the attending physician based on the type and severity of the disease being treated, the individual's health and medical history, concomitant medications, the specific compound being administered, and the route of administration. The dosage of the compounds of the present invention may be outside this dosage range, as needed.

[0229] It should be understood that within the scope of the present invention, the technical features defined in the above-mentioned technical solutions and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one.

[0230] Unless the structural formula is obviously incorrect, when the chemical name of any compound of the present invention is inconsistent with the given structural formula, the structural formula shall prevail.

[0231] As will be readily understood by those skilled in the art, for the sake of simplicity, not all hydrogen atoms are explicitly indicated in some of the compound formulas given herein. When there are vacant valences on carbon atoms or nitrogen atoms in a compound, this indicates the presence of unindicated hydrogen atoms. For example, the compound of Example P12 below is represented by the formula Description, wherein a hydrogen atom is omitted on the nitrogen atom between the pyrimidine ring and the pyrazole ring. It will be understood by those skilled in the art that this structural formula is similar to the structural formula represent the same compound. BRIEF DESCRIPTION OF THE DRAWINGS

[0232] Figure 1 Shown is a general synthetic scheme A for synthesizing compounds of the present invention, wherein the variables are as defined herein.

[0233] Figure 2 Shown is a general synthetic scheme B for synthesizing compounds of the present invention, wherein the variables are as defined herein.

[0234] Figure 3 Shown is a general synthetic scheme C for synthesizing compounds of the present invention, wherein the variables are as defined herein.

[0235] Figure 4 Shown is a general synthetic scheme D for synthesizing compounds of the present invention, wherein the variables are as defined herein. Example

[0236] The following examples are provided to further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0237] In the following examples, the experimental methods without specific conditions can be carried out according to the conventional conditions of such reactions or the conditions recommended by the manufacturers. Unless otherwise specified, percentages and parts are by weight.

[0238] Unless otherwise specified, the experimental materials and reagents used in the following examples can be obtained from commercial channels.

[0239] In the following examples, 1 H-NMR spectra were recorded using a Bluker AVHD 400 MHz or Bluker AVHD 500 MHz NMR spectra; 13 C-NMR spectra were recorded using a Bluker AVHD 500 MHz or Bluker AVHD 600 MHz nuclear magnetic resonance instrument, and chemical shifts are expressed in δ (ppm); mass spectra were recorded using a Waters UPLC H-Class+QDa (ESI) coupled with an Agilent 1260_6120 (ESI) mass spectrometer; and reversed-phase preparative HPLC separations were performed using a Waters UV-guided fully automated purification system (XBridge Prep C18 5 μm OBD column).

[0240] The abbreviations used in the examples have the following meanings:

[0241] iPrOH isopropyl alcohol

[0242] EtOH

[0243] DCM dichloromethane

[0244] TFA or CF3COOH trifluoroacetic acid

[0245] MeOH methanol

[0246] NaOH sodium hydroxide

[0247] HCl Hydrogen chloride or hydrochloric acid

[0248] TEA triethylamine Raney Ni

[0249] dioxane

[0250] NaH sodium hydride

[0251] H2O water

[0252] Pd / C Palladium / Carbon

[0253] H2 Hydrogen

[0254] N2 nitrogen HATU 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethyluronium hexafluorophosphate

[0255] DMF N,N-dimethylformamide

[0256] THF Tetrahydrofuran

[0257] Boc2O Di-tert-butyl dicarbonate

[0258] Boc tert-Butoxycarbonyl

[0259] NBS N-bromosuccinimide

[0260] NCS N-chlorosuccinimide

[0261] NIS N-iodosuccinimide

[0262] MeCN or CH3CN Acetonitrile

[0263] DIPEA or DIEA N,N-diisopropylethylamine

[0264] NaBH4 sodium borohydride

[0265] AcOH acetic acid

[0266] Ac2O acetic anhydride

[0267] AcCl Acetyl chloride NaBH3CN or NaBH3(CN) Sodium cyanoborohydride

[0268] K2CO3 potassium carbonate

[0269] Cs2CO3 cesium carbonate

[0270] NaHCO3 sodium bicarbonate

[0271] nBuLi n-butyllithium

[0272] LiAlH4 lithium aluminum hydride Pd(dppf)Cl2 or PdCl2(dppf) [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride

[0273] PdCl2(PPh3)2 Bistriphenylphosphine palladium dichloride

[0274] KOAc potassium acetate

[0275] Fumaronitrile

[0276] P(nBu)3 tri-n-butylphosphine

[0277] LDA lithium diisopropylamide

[0278] LiOH lithium hydroxide

[0279] MeI iodomethane

[0280] EtI Ethyl iodide

[0281] (CH2O) n Paraformaldehyde

[0282] HCO2H or FA Formic acid

[0283] CH3COCl Acetyl chloride

[0284] HPLC high-performance liquid chromatography

[0285] CH3COOK or AcOK Potassium Acetate

[0286] t-BuONa Sodium tert-butoxide

[0287] DMSO dimethyl sulfoxide

[0288] h hour

[0289] min

[0290] DMAP 4-dimethylaminopyridine

[0291] rt or RT room temperature

[0292] T3P 1-propylphosphoric anhydride

[0293] DMEA N,N-dimethylethanolamine

[0294] POCl3 phosphorus oxychloride

[0295] ℃ degrees Celsius

[0296] EA Ethyl acetate

[0297] Bu4NBr3 Tetrabutylammonium tribromide

[0298] CuI Cuprous iodide

[0299] Mg

[0300] Py pyridine

[0301] TLC thin layer chromatography

[0302] LCMS liquid chromatography-mass spectrometry

[0303] TBS tert-butyldimethylsilyl

[0304] TBSCl tert-Butyldimethylsilyl chloride

[0305] BPin2 Bis(pinacolato)diboron

[0306] PE petroleum ether

[0307] MW microwave

[0308] DEA Diethylamine

[0309] HEP n-heptane

[0310] IPA Isopropyl alcohol

[0311] HEX n-hexane

[0312] Synthesis of intermediate 4 :Synthesis of 7-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indoline (4)

[0313]

[0314] Step 1. Synthesis of 7-fluoroindoline (2)

[0315]

[0316] Compound 1 (5000 mg, 37 mmol) and AcOH (30 mL) were added to a dry 50 mL round-bottom flask at room temperature. NaBH3CN (5813 mg, 92.49 mmol) was added portionwise at 0°C and stirred at room temperature for 3 hours. After completion of the reaction, the mixture was concentrated under reduced pressure and 100 mL of water was added. The pH was adjusted to 9 with 2 mol / L aqueous sodium hydroxide at 0°C and stirred for 1 hour. The mixture was then heated to 25°C and stirred for 1 hour. The mixture was extracted with ethyl acetate, dried, filtered, and the filtrate was evaporated to dryness. The resulting residue was purified by silica gel column chromatography using an eluent system (ethyl acetate / petroleum ether = 1 / 50 to 1 / 10) to afford intermediate 2 (7-fluoroindoline) as an off-white solid (4.0 g) in a 78.4% yield. LCMS: m / z 138.1 (M+H).

[0317] Step 2. Synthesis of 5-bromo-7-fluoroindoline (3)

[0318]

[0319] Method 1:

[0320] To a solution of Intermediate 2 (4.0 g, 29.19 mmol) in acetonitrile (100 mL) in a dry 250 mL round-bottom flask at 0°C, NBS (5.2 g, 29.19 mmol) was slowly added. The mixture was then warmed to room temperature and stirred for 2 hours. After completion, the reaction was concentrated under reduced pressure, 100 mL of water was added, and the mixture was extracted with ethyl acetate (150 mL x 3). The organic phases were combined. The mixture was washed with saturated sodium bicarbonate aqueous solution and saturated brine (100 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using an eluent system (ethyl acetate:petroleum ether = 1:8) to afford Intermediate 3 (5.0 g, pale purple solid) in an 80.6% yield. LCMS: m / z 215.9 (M+H).

[0321] Method 2:

[0322] Compound 2 (9.8 g, 0.072 mol) and dichloromethane (100 mL) were added to a 100 mL flask in an ice-water bath. Tetrabutylammonium tribromide (34.5 g, 0.072 mol) was then added portionwise. The mixture was allowed to warm to room temperature and stirred for 4 hours. After completion, the reaction was concentrated under reduced pressure. Saturated aqueous sodium bicarbonate was slowly added to adjust the pH to 6-7. The mixture was extracted with ethyl acetate (50 mL x 4). The organic phases were combined and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using an eluent system (ethyl acetate:petroleum ether = 1:15) to yield 5-bromo-7-fluoroindoline 3 (9.5 g, off-white solid). LCMS: m / z 215.9 (M+H).

[0323] Step 3. Synthesis of 7-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-indoline (4)

[0324]

[0325] To a dry 50 mL single-necked bottle at room temperature were added intermediate 3 (600 mg, 2.777 mmol), bis(pinacolato)diboron (1410 mg, 5.554 mmol), Pd(dppf)Cl2 (243 mg, 0.333 mmol), potassium acetate (545 mg, 5.554 mmol), and 1,4-dioxane (4 mL). The atmosphere was purged with nitrogen three times. The mixture was stirred and heated to 110°C for 3 hours. After completion of the reaction, as determined by LCMS, the mixture was filtered, the filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography using an eluent system (EA / PE = 2%-10%) to afford the intermediate 4,7-fluoro-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-2,3-dihydro-1H-indole (600 mg, yellow solid) in 82.1% yield and approximately 90% purity. LCMS: m / z 264.1 (M+H).

[0326] Intermediate 7 :Synthesis of 4-bromo-N-(1-methyl-1H-pyrazol-5-yl)pyridin-2-amine (7)

[0327]

[0328] To a dry 250 mL round-bottom flask at room temperature, compound 6 (4.2 g, 23.86 mmol), compound 5 (2.8 g, 28.83 mmol), sodium tert-butoxide (4.6 g, 47.8 mmol), and DMSO (60 mL) were added. The atmosphere was replaced with nitrogen three times, and the mixture was stirred and refluxed at 125°C for 24 hours. After completion of the reaction, the mixture was extracted with EA (50 mL x 3), washed with saturated brine (30 mL), dried, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using an eluent system (PE:EA = 3:7) to afford intermediate 7 (2.1 g, pale yellow solid) in a 40% yield. LCMS: m / z 253.0 / 255.0 (M+H).

[0329] Intermediate 8 :Synthesis of tert-butyl (4-bromopyridin-2-yl)(1-methyl-1H-pyrazol-5-yl)carbamate (8)

[0330]

[0331] To a dry 100 mL round-bottom flask at room temperature, intermediate 7 (500 mg, 1.976 mmol), DMAP (67 mg, 0.593 mmol), DIPEA (766 mg, 5.928 mmol), and DMF (5 ml) were added. After stirring at room temperature to dissolve, Boc2O (1299 mg, 5.928 mmol) was slowly added and allowed to react at room temperature for 3 hours. After completion of the reaction, the mixture was extracted with EA, and the organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting mixture was purified by silica gel column chromatography using an eluent system (ethyl acetate / petroleum ether = 15 / 85 to 70 / 30) to obtain intermediate 8 (626 mg, light yellow solid) in an 88% yield. LCMS: m / z 352.9 (M+H).

[0332] Intermediate 13 :Synthesis of 1-(5-(2-bromopyrimidin-4-yl)-7-fluoroindolin-1-yl)-2-(2-chloropyridin-3-yl)ethan-1-one (13)

[0333]

[0334] Step 1. Synthesis of 1-(5-bromo-7-fluoroindolin-1-yl)-2-(2-chloropyridin-3-yl)ethan-1-one (10)

[0335]

[0336] To a dry 250 mL round-bottom flask at room temperature, N,N-dimethylformamide (10 mL), intermediate 3 (1.7 g, 0.0079 mol), compound 9 (1.23 g, 0.007 mol), and N,N-diisopropylethylamine (3.69 g, 0.028 mol) were added. The atmosphere was purged with nitrogen three times, and the temperature was gradually raised to 50°C. 1-propylphosphoric anhydride (56 mL, 50% ethyl acetate solution) was added and allowed to react for 0.5 hours. After completion of the reaction, as determined by TLC, the product was concentrated under reduced pressure and the residue poured into ice water to precipitate a solid, which was filtered to afford intermediate 10 (2.7 g, light yellow solid) in a 93% yield. LCMS: m / z 368.8 / 370.8 (M+H).

[0337] Step 2. Synthesis of 2-(2-chloropyridin-3-yl)-1-(7-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-1-yl)ethan-1-one (11)

[0338]

[0339] To a dry 100 mL round-bottom flask at room temperature, Intermediate 10 (2 g, 0.0054 mol), bis(pinacolato)diboron (2.0 g, 0.0079 mol), potassium acetate (1.59 g, 0.016 mol), and 1,4-dioxane (36 mL) were added. The atmosphere was purged with nitrogen once, and Pd(dppf)Cl2 (442 mg, 0.0005 mol) was added. The atmosphere was purged with nitrogen three times, and the mixture was heated to 90°C and stirred for 4 hours. After completion of the reaction, as monitored by LCMS, the mixture was concentrated under reduced pressure and the resulting residue was purified by silica gel column chromatography with an eluent system (ethyl acetate:petroleum ether = 1:3) to afford Intermediate 11 (1.5 g, light yellow solid) in a 68% yield. LCMS: m / z 417.1 (M+H).

[0340] Step 3. Synthesis of 1-(5-(2-bromopyrimidin-4-yl)-7-fluoroindolin-1-yl)-2-(2-chloropyridin-3-yl)ethan-1-one (13)

[0341]

[0342] To a dry 100 mL round-bottom flask at room temperature were added intermediate 11 (200 mg, 0.48 mmol), 2,4-dibromopyrimidine (126 mg, 0.53 mmol), potassium carbonate (200 mg, 1.44 mmol), (1,1-bis(diphenylphosphino)ferrocene)palladium dichloride (70 mg, 0.096 mmol), and a 4:1 mixture of 1,4-dioxane and water (10.0 mL). The atmosphere was replaced with nitrogen three times, the temperature was raised to 60°C, and the mixture was stirred for 2 hours. After completion of the reaction, the mixture was concentrated under reduced pressure, 100 mL of water was added, and the mixture was extracted with ethyl acetate (150 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using an eluent system (ethyl acetate:petroleum ether = 1:1) to afford product 13 (140 mg, pale yellow solid) in a 65.0% yield. LCMS: m / z 446.8 (M+H).

[0343] Intermediate 17 :Synthesis of 4-chloro-N-(1-methyl-1H-pyrazol-5-yl)pyrimidin-2-amine (17)

[0344]

[0345] Step 1. Synthesis of 2-(1-methyl-1H-pyrazol-5-yl)amino-pyrimidin-4(3H)-one (16)

[0346]

[0347] To a dry 50 mL round-bottom flask at room temperature, compound 14 (7.8 g, 80 mmol), compound 15 (5.72 g, 40 mmol), and trimethylacetic acid (28.6 g) were added. Stirring was initiated and the mixture was slowly heated to 150°C for 40 hours. After completion of the reaction, the mixture was cooled to room temperature and 30 mL of dichloromethane and 5 mL of methanol were added to dissolve the reaction mixture completely. Silica gel was added and the sample was mixed. Intermediate 16 (5 g, yellow solid) was purified by silica gel column chromatography using an eluent system (dichloromethane:methanol = 3:2) to obtain intermediate 16 (5 g, yellow solid) in a 65.8% yield. LCMS: m / z 191.9 (M+H).

[0348] Step 2. Synthesis of 4-chloro-N-(1-methyl-1H-pyrazol-5-yl)pyrimidin-2-amine (17)

[0349]

[0350] To a dry 250 mL round-bottom flask at room temperature, intermediate 16 (5 g, 26.1 mmol), phosphorus oxychloride (10 mL, 109.2 mmol), and acetonitrile (100.0 mL) were added. The mixture was heated to 100°C and stirred for 2 hours. After completion, the reaction was concentrated under reduced pressure, quenched with 100 mL of water, and extracted with ethyl acetate (60 mL x 3). The organic phases were combined and washed with saturated brine (40 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using an eluent system (ethyl acetate:petroleum ether = 3:2) to afford intermediate 17 (2.9 g, light yellow solid) in a 53.7% yield. LCMS: m / z 209.9 (M+H).

[0351] 1 H-NMR (CDCl3, 400MHz): 8.27 (d, J = 5.2Hz, 1H), 7.53 (br, 1H), 7.48 (d, J = 5.2Hz, 1H), 6.80 (d, J = 5.6Hz, 1H), 6.28 (d, J = 5.6Hz, 1H), 3.77 (s, 3H).

[0352] Intermediate 23 :Synthesis of 1-(7-chloro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-1-yl)-2-(2-chloropyridin-3-yl)ethan-1-one (23)

[0353]

[0354] Step 1. Synthesis of 1-(5-bromo-dihydroindolin-1-yl)ethan-1-one (19)

[0355]

[0356] To a dry 50 mL round-bottom flask at room temperature, compound 18 (1500 mg, 7.57 mmol), DCM (20 mL), Et3N (1915 mg, 18.93 mmol), and DMAP (30 mg) were added. Acetyl chloride (1188 mg, 15.14 mmol) was added portionwise at 0°C and stirred at room temperature for 3 hours. The mixture was poured into water at 0°C and stirred for 1 hour. Extraction with DCM, drying, filtration, and evaporation to dryness afforded Intermediate 19 (1.75 g, yellow solid) in a 96.3% yield. LCMS: m / z 240.0 / 242.0 (M+H).

[0357] Step 2. 1-(5-Bromo-7-chloro-dihydroindolin-1-yl)ethan-1-one (20)

[0358]

[0359] To a dry 50 mL round-bottom flask at room temperature, Intermediate 19 (1600 mg, 14.58 mmol), acetonitrile (30 mL), and NCS (979 mg, 7.33 mmol) were added. After complete addition, the mixture was stirred at 70°C for 18 hours. The mixture was poured into water (20 mL), extracted with EA, dried, filtered, and the filtrate evaporated to dryness. The resulting residue was purified by silica gel column chromatography using an eluent system (ethyl acetate / petroleum ether = 1 / 50 to 1 / 10) to afford Intermediate 20 (1.2 g, yellow liquid) in a 65.6% yield. LCMS: m / z 274.0 / 276.0 (M+H).

[0360] Step 3. Synthesis of 5-bromo-7-chloro-dihydroindoline (21)

[0361]

[0362] To a dry 50 mL single-necked bottle at room temperature, intermediate 20 (1200 mg, 4.37 mmol), lithium hydroxide monohydrate (550 mg, 13.11 mmol), methanol (15 mL), and water (8 mL) were added. The mixture was stirred and heated to 70°C for 18 hours. After completion of the reaction, as determined by LCMS, the product was extracted with EA, washed, dried, and filtered. The filtrate was concentrated under reduced pressure and purified by silica gel column chromatography using an eluent system (EA / PE = 2%-10%) to afford intermediate 21 (700 mg, yellow solid) in a 68.9% yield with an approximate purity of 80%. LCMS: m / z 232.0 (M+H).

[0363] Step 4. Synthesis of 7-chloro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-indoline (22)

[0364]

[0365] To a dry 50 mL single-necked vial at room temperature were added Intermediate 21 (600 mg, 2.581 mmol), bis(pinacolato)diboron (852 mg, 3.354 mmol), Pd(dppf)Cl2 (188 mg, 0.2581 mmol), potassium acetate (329 mg, 3.354 mmol), and 1,4-dioxane (5 mL). The atmosphere was purged with nitrogen three times. The mixture was stirred and heated to 100°C for 3 hours. After completion of the reaction, as determined by LCMS, the mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using an eluent system (EA / PE = 2%-10%) to afford Intermediate 22 (480 mg, yellow solid) in a 66.67% yield with a purity of approximately 80%. LCMS: m / z 280.1 (M+H).

[0366] Step 5. Synthesis of intermediate 1-(7-chloro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-1-yl)-2-(2-chloropyridin-3-yl)ethan-1-one (23)

[0367]

[0368] To a dry 25 mL single-necked vial at room temperature, intermediate 22 (440 mg, 1.574 mmol), compound 9 (540 mg, 3.147 mmol), Et3N (318 mg, 3.147 mmol), HATU (1193 mg, 3.147 mmol), and THF (8 mL) were added. The mixture was stirred at 32°C under nitrogen for 18 hours. After completion of the reaction, as determined by LCMS, the mixture was extracted with EA, washed with water, dried, and evaporated to dryness. The crude product was purified by silica gel column chromatography using an eluent system (EA / PE 10-40%) to afford intermediate 23 (290 mg, yellow solid) in a 42.58% yield. LCMS: m / z 434.1 (M+H).

[0369] Intermediate 31 :Synthesis of 2-(2-chloropyridin-3-yl)-1-(7-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-1-yl)ethan-1-one (31)

[0370]

[0371] Step 1. Synthesis of 7-methylindoline (28)

[0372]

[0373] To a dry 250 mL round-bottom flask at 0°C, compound 27 (7-methyl-1H-indole) (2.0 g, 15.27 mmol), sodium cyanoborohydride (2.9 g, 45.81 mmol), and glacial acetic acid (40 mL) were added. The mixture was allowed to warm to room temperature and stirred for 3 hours. After completion of the reaction, the mixture was concentrated under reduced pressure, 100 mL of water was added, and the pH was adjusted to 9 with 2 mol / L aqueous sodium hydroxide. The mixture was extracted with ethyl acetate (100 mL x 3). The combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by flash column chromatography using an eluent system (ethyl acetate:petroleum ether = 1:5) to afford intermediate 28 (7-methylindoline) as an off-white solid (1.5 g) in a 75.3% yield. LCMS: m / z 134.1 (M+H).

[0374] Step 2. Synthesis of 5-bromo-7-methylindoline (29)

[0375]

[0376] To a dry 100 mL round-bottom flask at room temperature, Intermediate 28 (7-methylindoline) (120 mg, 0.902 mmol), tetrabutylammonium tribromide (464 mg, 0.902 mmol), and dichloromethane (10.0 mL) were added and stirred for 30 minutes. After completion of the reaction, as monitored by LCMS, the mixture was concentrated under reduced pressure, 100 mL of water was added, and extraction with ethyl acetate (150 mL x 3) was performed. The combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using an eluent system (ethyl acetate:petroleum ether = 1:5) to afford Intermediate 29 (5-bromo-7-methylindoline) as a pale purple solid (200 mg) in an 80.6% yield. LCMS: m / z 211.9 (M+H).

[0377] Step 3. Synthesis of 1-(5-bromo-7-methylindolin-1-yl)-2-(2-chloropyridin-3-yl)ethan-1-one (30)

[0378]

[0379] In a dry 100 mL round-bottom flask, intermediate 29 (80 mg, 0.38 mmol), compound 9 2-(2-chloropyridin-3-yl)acetic acid (67 mg, 0.38 mmol), N,N-diisopropylethylamine (0.25 mL, 1.52 mmol), T3P (1.208 g, 50% (wt%) ethyl acetate solution (1.9 mmol)) and N,N-dimethylformamide (5.0 mL) were added and the reaction was stirred at room temperature for 30 minutes. After completion of the reaction, 100 mL of water was added, and the mixture was extracted with ethyl acetate (150 mL × 3). The combined organic phases were washed with saturated brine (100 mL × 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with an eluent system (ethyl acetate:petroleum ether = 1:5) to obtain Intermediate 30, 1-(5-bromo-7-methylindolin-1-yl)-2-(2-chloropyridin-3-yl)ethan-1-one (50 mg, off-white solid) in a 36.3% yield. LCMS: m / z 364.8 (M+H).

[0380] Step 4. Synthesis of 2-(2-chloropyridin-3-yl)-1-(7-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-1-yl)ethan-1-one (31)

[0381]

[0382] To a dry 100 mL round-bottom flask were added intermediate 30 1-(5-bromo-7-methylindolin-1-yl)-2-(2-chloropyridin-3-yl)ethan-1-one (400 mg, 1.09 mmol), bis(pinacolato)diboron (1.38 g, 5.43 mmol), potassium acetate (233 mg, 3.27 mmol), (1,1-bis(diphenylphosphino)ferrocene)palladium dichloride (70 mg, 0.11 mmol) and 1,4-dioxane (20.0 mL) at room temperature. The atmosphere was replaced with nitrogen five times, the temperature was raised to 90°C, and the reaction was stirred overnight. After completion of the reaction, as monitored by LCMS, the reaction mixture was concentrated under reduced pressure, 100 mL of water was added, and the mixture was extracted with ethyl acetate (150 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with an eluent system (ethyl acetate:petroleum ether = 1:10) to provide Intermediate 31, 2-(2-chloropyridin-3-yl)-1-(7-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-1-yl)ethan-1-one (200 mg, white solid), in a 44.0% yield. LCMS: m / z 413.0 (M+H).

[0383] Intermediate 53:Synthesis of 2-(2-chloropyridin-3-yl)-1-(7-fluoro-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-1-yl)ethan-1-one

[0384]

[0385] Step 1. Synthesis of ethyl 2-(3-fluoro-2-nitrophenyl)-3-oxobutanoate (47)

[0386]

[0387] To a dry 100 mL round-bottom flask at room temperature, ethyl acetoacetate (46) (6.1 g, 0.047 mol), potassium carbonate (8.7 g, 0.063 mol), 1,3-difluoro-2-nitrobenzene (5 g, 0.031 mol), and N,N-dimethylformamide (20 mL) were added in sequence. The temperature was raised to 50°C and the reaction was stirred for 2 hours. After the reaction was completed, it was diluted with water (100 mL) and extracted with ethyl acetate (80 mL × 3). The organic phases were combined, washed with saturated brine (300 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 1:100) to obtain ethyl 2-(3-fluoro-2-nitrophenyl)-3-oxobutanoate 47 (2.8 g, light yellow oil) in a yield of 33%. 1 H NMR(CDCl3,300MHz):13.07(s,1H),7.58-7.40(m,1H),7.30-7.19(m,1H),7.16-7.01(m,1H),4.31-4.00(m,2H),1.87(s,3H).1.18(t,J=7.2Hz,3H).

[0388] Step 2. Synthesis of 1-(3-fluoro-2-nitrophenyl)propan-2-one (48)

[0389]

[0390] To a dry 100 mL round-bottom flask at room temperature, ethyl 2-(3-fluoro-2-nitrophenyl)-3-oxobutanoate (2.8 g, 0.01 mol), acetic acid (20 mL), and 50% sulfuric acid (20 mL) were added. The temperature was raised to 100°C and the reaction was stirred for 4 hours. After completion of the reaction, as monitored by TLC, the mixture was diluted with water (100 mL) and extracted with ethyl acetate (80 mL × 3). The organic phases were combined, washed with saturated sodium bicarbonate (200 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography (ethyl acetate:petroleum ether = 1:5) to obtain 1-(3-fluoro-2-nitrophenyl)propan-2-one 48 (1.86 g, light yellow oil) in a 91% yield. 1 H NMR (DMSO-d6, 300MHz): 7.73-7.63 (m, 1H), 7.57-7.48 (m, 1H), 7.31 (d, J = 7.5Hz, 1H), 4.13 (s, 3H), 2.51 (s, 2H).

[0391] Step 3. Synthesis of 7-fluoro-2-methyl-1H-indole (49)

[0392]

[0393] To a dry 100 mL round-bottom flask, distilled water (32 mL) and sodium dithionite (13.2 g, 0.076 mol) were added sequentially. A solution of 1-(3-fluoro-2-nitrophenyl)propan-2-one (1.5 g, 0.0076 mmol) in 1,4-dioxane (3.4 mL) was then added dropwise. The mixture was stirred at room temperature for 2 hours. After completion of the reaction, as monitored by TLC, the mixture was diluted with water (80 mL) and extracted with ethyl acetate (60 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. Purification by silica gel column chromatography (pure petroleum ether) afforded 7-fluoro-2-methyl-1H-indole 49 (250 mg, white solid) in a 22% yield. 1 H NMR (DMSO-d6, 300MHz): 11.34 (s, 1H), 7.21 (d, J = 6.9Hz, 1H), 6.93-6.72 (m, 2H), 6.19 (s, 1H), 2.39 (s, 3H).

[0394] Step 4. Synthesis of 7-fluoro-2-methylindoline (50)

[0395]

[0396] In a dry 100 mL round-bottom flask, 7-fluoro-2-methyl-1H-indole 49 (1.8 g, 12.08 mmol), acetic acid (20 mL), and sodium cyanoborohydride (2.28 g, 36.19 mmol) were added sequentially under ice. The mixture was slowly warmed to room temperature and stirred for 2 hours. After completion of the reaction, the mixture was concentrated under reduced pressure. The resulting residue was dissolved in ethyl acetate (100 mL), washed with saturated sodium bicarbonate (80 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 1:70) to afford 7-fluoro-2-methylindoline 50 (1.4 g, colorless oil) in a 77% yield. LCMS: m / z 152.1 (M+H).

[0397] Step 5. Synthesis of 5-bromo-7-fluoro-2-methylindoline (51)

[0398]

[0399] To a dry 100 mL round-bottom flask in an ice bath, 7-fluoro-2-methylindoline 50 (1.4 g, 9.27 mmol), acetonitrile (20 mL), and N-bromosuccinimide (1.65 g, 9.27 mmol) were added sequentially. The mixture was gradually warmed to room temperature and stirred for 2 hours. After completion of the reaction, the mixture was concentrated under reduced pressure and purified by column chromatography (ethyl acetate:petroleum ether = 1:70) to afford 5-bromo-7-fluoro-2-methylindoline 51 (1.5 g, light yellow oil) in a 71% yield. LCMS: m / z 229.9 / 231.9 (M+H).

[0400] Step 6. Synthesis of 1-(5-bromo-7-fluoro-2-methylindolin-1-yl)-2-(2-chloropyridin-3-yl)ethan-1-one (52)

[0401]

[0402] To a dry 100 mL round-bottom flask at room temperature were added 5-bromo-7-fluoro-2-methylindoline 7 (1.0 g, 4.35 mmol), 2-(2-chloropyridin-3-yl)acetic acid (748 mg, 4.35 mmol), N,N-dimethylformamide (10 mL), and N,N-diisopropylethylamine (2.24 g, 17.36 mmol). The atmosphere was purged with nitrogen three times and the temperature was raised to 50°C. 1-propylphosphoric anhydride (26 mL, 50% ethyl acetate solution) was added and stirred for 0.5 h. After completion of the reaction, as monitored by TLC, the reaction was concentrated under reduced pressure and the residue poured into ice water to precipitate a solid, which was filtered and dried to afford 1-(5-bromo-7-fluoro-2-methylindoline-1-yl)-2-(2-chloropyridin-3-yl)ethan-1-one 52 (1.5 g, light brown solid) in a 90% yield. LCMS: m / z 382.8 / 384.8 (M+H).

[0403] Step 7. Synthesis of 2-(2-chloropyridin-3-yl)-1-(7-fluoro-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-1-yl)ethan-1-one (53)

[0404]

[0405] To a dry 100 mL round-bottom flask at room temperature, 1-(5-bromo-7-fluoro-2-methylindolin-1-yl)-2-(2-chloropyridin-3-yl)ethan-1-one (52) (500 mg, 1.3 mmol), bis(pinacolato)diboron (496 mg, 1.95 mmol), potassium acetate (383 mg, 3.9 mmol) and 1,4-dioxane (20 mL) were added in sequence. The nitrogen atmosphere was replaced once, and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (10.6 mg, 0.013 mol) was added. The nitrogen atmosphere was replaced three times. The mixture was heated to 90°C and stirred for 3 hours. After completion of the reaction, as monitored by LCMS, the reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by column chromatography (ethyl acetate:petroleum ether = 1:3) to provide 2-(2-chloropyridin-3-yl)-1-(7-fluoro-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-1-yl)ethan-1-one 53 (450 mg, colorless oil) in an 80% yield. LCMS: m / z 430.9 (M+H). Intermediate 70 :Synthesis of 2-(2-chloropyridin-3-yl)-1-(4,7-difluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-1-yl)ethan-1-one (70)

[0406]

[0407] Step 1. Synthesis of 5-bromo-4,7-difluoro-1H-indole (67)

[0408]

[0409] Vinylmagnesium bromide (45.6 mL, 45.6 mmol) was added to tetrahydrofuran (50 mL), and the mixture was cooled to -78°C under nitrogen. Compound 66 (3.9 g, 15.2 mmol, dissolved in 50 mL of tetrahydrofuran) was added dropwise. After the addition, the mixture was stirred for 2 hours. After completion of the reaction, saturated aqueous ammonium chloride (20 mL) was slowly added, and the mixture was extracted with ethyl acetate (20 mL × 3). The organic phases were combined and concentrated under reduced pressure. The residue was purified by silica gel column chromatography with an eluent system (ethyl acetate:petroleum ether = 1:8) to give 5-bromo-4,7-difluoro-1H-indole 67 (1.47 g, light yellow oil) in a yield of 38.7%.

[0410] 1 H NMR (CD3OD, 300MHz): 7.29 (d, J = 3.0Hz, 1H), 7.01-6.97 (m, 1H), 6.60-6.54 (m, 1H).

[0411] Step 2. Synthesis of 5-bromo-4,7-difluoro-dihydroindoline (68)

[0412]

[0413] To a 50 mL round-bottom flask in an ice-water bath, compound 67 (1.45 g, 6.28 mmol) and glacial acetic acid (20 mL) were added sequentially. Sodium cyanoborohydride (0.87 g, 12.6 mol) was added portionwise over approximately 30 minutes and stirred at room temperature for 16 hours. After completion of the reaction, the mixture was cooled in an ice-water bath and slowly adjusted to pH 6-7 with saturated sodium bicarbonate solution. The mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated. The resulting residue was purified by silica gel column chromatography using an eluent system (ethyl acetate:petroleum ether = 1:6) to afford 5-bromo-4,7-difluoro-indoline 68 (0.43 g, pale yellow oil) in a 30.0% yield. LCMS: m / z 234.0 / 236.0 (M+H).

[0414] Step 3. Synthesis of 1-(5-bromo-4,7-difluoro-indolin-1-yl)-2-(2-chloropyridin-3-yl)ethan-1-one (69)

[0415]

[0416] To a 100 mL flask were added compound 68 (0.43 mg, 0.1.85 mol), ethyl acetate (10 mL), compound 9 (0.35 g, 2.04 mmol), T3P (50% W / W solution in ethyl acetate, 2.83 g, 3.71 mmol), and diisopropylethylamine (0.48 g, 3.71 mmol) in sequence at room temperature. The mixture was stirred at room temperature for 3 hours. After completion of the reaction, saturated aqueous sodium bicarbonate solution was slowly added to adjust the pH to 6-7. The mixture was extracted with ethyl acetate (10 mL x 4). The organic phases were combined, concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with an eluent system (ethyl acetate:petroleum ether = 1:3) to afford 1-(5-bromo-4,7-difluoro-indolin-1-yl)-2-(2-chloropyridin-3-yl)ethan-1-one 69 (0.56 g, light yellow oil) in a yield of 78.2%. LCMS: m / z 386.9 / 388.9 (M+H).

[0417] Step 4. Synthesis of 2-(2-chloropyridin-3-yl)-1-(4,7-difluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-1-yl)ethan-1-one (70)

[0418]

[0419] In a dry 100 mL round-bottom flask, compound 69 (0.27 g, 0.69 mmol), bis(pinacolato)diboron (0.23 g, 0.89 mol), potassium acetate (0.10 g, 1.02 mmol), and (1,1'-bis(diphenylphosphino)ferrocene)palladium dichloride (0.028 g, 0.89 mmol) were added to 1,4-dioxane (10 mL) in sequence. The atmosphere was replaced with nitrogen three times, the temperature was raised to 90°C, and the reaction was stirred for 16 hours. After completion of the reaction, the mixture was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with an eluent system (ethyl acetate:petroleum ether = 1:3) to afford crude 2-(2-chloropyridin-3-yl)-1-(4,7-difluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-1-yl)ethan-1-one 70 (0.18 g, pale yellow solid). LCMS: m / z 434.7 / 436.8 (M+H).

[0420] Intermediate 77: Synthesis of 2-(2-chloropyridin-3-yl)-1-(6,7-difluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-1-yl)ethan-1-one (77)

[0421]

[0422] Step 1. Synthesis of 4-bromo-2,3-difluoro-6-iodoaniline (72)

[0423]

[0424] Compound 71 (0.85 g, 4.1 mmol), glacial acetic acid (15 mL), and N-iodosuccinimide (0.97 g, 4.31 mmol) were added sequentially to a 100 mL round-bottom flask. The mixture was stirred at room temperature for 3 hours. After completion of the reaction, the mixture was concentrated under reduced pressure. Saturated aqueous sodium bicarbonate solution was added to the residue to adjust the pH to 6-7. The mixture was extracted with ethyl acetate (20 mL × 3). The organic phases were combined and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using an eluent system (ethyl acetate:petroleum ether = 1:10) to give 4-bromo-2,3-difluoro-6-iodoaniline 72 (1.30 g, light yellow solid) in a 95% yield. LCMS: m / z 333.9 / 335.8 (M+H). Step 2. Synthesis of 4-bromo-2,3-difluoro-6-((trimethylsilyl)ethynyl)aniline (73)

[0425]

[0426] To a 100 mL round-bottom flask were added compound 72 (1.30 g, 3.91 mmol), triethylamine (20 mL), cuprous iodide (37.2 mg, 0.20 mol), ethynyltrimethylsilane (460 mg, 4.69 mmol), and bistriphenylphosphine palladium dichloride (137 mg, 0.20 mmol). The mixture was stirred at room temperature for 16 hours. After completion of the reaction, the mixture was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography using an eluent system (ethyl acetate:petroleum ether = 1:10) to provide 4-bromo-2,3-difluoro-6-((trimethylsilyl)ethynyl)aniline 73 (1.02 g, light yellow oil) in an 86.2% yield. 1 H NMR (DMSO-d6, 400MHz): δ7.32 (d, J = 7.2, 1H), 5.86 (s, 2H), 0.25 (s, 9H).

[0427] Step 3. Synthesis of 5-bromo-6,7-difluoro-1H-indole (74)

[0428]

[0429] Compound 73 (0.85 g, 2.80 mmol), N,N-dimethylformamide (2 mL), and cuprous iodide (1.07 g, 5.61 mmol) were added to a 50 mL eggplant-shaped flask. The mixture was heated to 100°C under nitrogen and stirred for 4 hours. After completion of the reaction, the mixture was cooled to room temperature, and water (20 mL) was added. The mixture was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with an eluent system (ethyl acetate:petroleum ether = 1:6) to afford 5-bromo-6,7-difluoro-1H-indole 74 (560 mg, light yellow oil) in an 86% yield. 1 H NMR (DMSO-d6, 400MHz): δ12.46 (s, 1H), 7.74-7.70 (m, 1H), 7.51 (s, 1H), 3.36 (d, J = 5.6Hz, 1H).

[0430] Step 4. Synthesis of 5-bromo-6,7-difluoro-indoline (75)

[0431]

[0432] To a 50 mL round-bottom flask in an ice-water bath, compound 74 (0.56 g, 2.42 mmol) and glacial acetic acid (10 mL) were added sequentially. Sodium cyanoborohydride (0.35 g, 4.85 mmol) was added portionwise over approximately 30 minutes. The reaction was stirred at room temperature for 16 hours. After completion of the reaction, the mixture was placed in an ice-water bath. Saturated sodium bicarbonate solution was slowly added to adjust the pH to 6-7. The mixture was then extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The resulting residue was purified by silica gel column chromatography using an eluent system (ethyl acetate:petroleum ether = 1:6) to afford 5-bromo-6,7-difluoro-dihydroindoline 75 (0.23 g) as a pale yellow crude oil. LCMS: m / z 233.9 / 235.9 (M+H); RT = 1.470 min (2.5 min).

[0433] Step 5. Synthesis of 1-(5-bromo-6,7-difluoro-indolin-1-yl)-2-(2-chloropyridin-3-yl)ethan-1-one (76)

[0434]

[0435] To a 100 mL flask were added compound 75 (0.23 mg, 0.99 mmol), ethyl acetate (10 mL), compound 9 (0.20 g, 1.18 mmol), T3P (50% W / W solution in ethyl acetate, 1.26 g, 1.98 mmol), and diisopropylethylamine (0.38 g, 2.96 mmol) in sequence at room temperature. The mixture was stirred at room temperature for 3 hours. After completion of the reaction, saturated aqueous sodium bicarbonate solution was slowly added to adjust the pH to 6-7. The mixture was extracted with ethyl acetate (10 mL x 4). The organic phases were combined, concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with an eluent system (ethyl acetate:petroleum ether = 1:3) to afford 1-(5-bromo-6,7-difluoro-indolin-1-yl)-2-(2-chloropyridin-3-yl)ethan-1-one 76 (0.10 g, light yellow oil) in a yield of 26.2%. LCMS: m / z 386.9 / 388.9 (M+H); RT=1.427min (2.5min).

[0436] Step 6. Synthesis of 2-(2-chloropyridin-3-yl)-1-(6,7-difluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-1-yl)ethan-1-one (77)

[0437]

[0438] In a dry 50 mL round-bottom flask, compound 76 (0.10 g, 0.26 mmol), bis(pinacolato)diboron (0.079 g, 0.31 mmol), potassium acetate (0.038 g, 0.39 mmol), and (1,1'-bis(diphenylphosphino)ferrocene)palladium dichloride (0.011 g, 0.013 mmol) were added to 1,4-dioxane (10 ml) in sequence. The atmosphere was replaced with nitrogen three times, the temperature was raised to 90°C, and the reaction was stirred for 5 hours. After completion of the reaction, the mixture was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with an eluent system (ethyl acetate:petroleum ether = 1:3) to afford crude 2-(2-chloropyridin-3-yl)-1-(6,7-difluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-1-yl)ethan-1-one 77 (0.065 g, pale yellow solid). LCMS: m / z 435.1 / 437.1 (M+H).

[0439] Intermediate 89: Synthesis of 2-(2-chloropyridin-3-yl)-1-(7-fluoro-3-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-1-yl)ethan-1-one (89)

[0440]

[0441] Step 1. Synthesis of 7-fluoro-3-methyl-1H-indole (85)

[0442]

[0443] Compound 84 (200.0 mg, 1.23 mmol) was added to tetrahydrofuran (15 mL), the atmosphere was purged with nitrogen three times, and the mixture was cooled to 0°C. Lithium aluminum hydride (140.0 mg, 3.69 mmol) was added. After the addition was complete, the mixture was allowed to warm to room temperature and stirred for 2 hours. After completion of the reaction, water (20 mL) was slowly added dropwise, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phases were combined and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using an eluent system (ethyl acetate:petroleum ether = 1:8) to afford 7-fluoro-3-methyl-1H-indole 85 (130.0 mg, light yellow oil) in a yield of 71.0%.

[0444] 1 H NMR (DMSO-d6, 400MHz): δ11.21 (s, 1H), 7.29 (d, J = 7.6Hz, 1H), 7.17 (s, 1H), 6.97-6.87 (m, 2H).

[0445] Step 2. Synthesis of 7-fluoro-3-methyl-dihydroindoline (86)

[0446]

[0447] To a 50 mL round-bottom flask in an ice-water bath, compound 85 (819.0 mg, 5.49 mmol) and glacial acetic acid (20 mL) were added sequentially. Sodium cyanoborohydride (691.0 mg, 10.99 mol) was added portionwise over approximately 30 minutes. The mixture was allowed to warm to room temperature and stirred for 16 hours. After completion of the reaction, the mixture was cooled in an ice-water bath. A saturated sodium bicarbonate solution was slowly added to adjust the pH to 6-7. The mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The resulting residue was purified by silica gel column chromatography using an eluent system (ethyl acetate:petroleum ether = 1:6) to afford 7-fluoro-3-methyl-dihydroindoline 86 (49.0 mg, pale yellow oil). LCMS: m / z 152.0 (M+H).

[0448] Step 3. Synthesis of 5-bromo-7-fluoro-3-methyl-dihydroindoline (87)

[0449]

[0450] In a 50 mL flask under an ice-water bath, compound 86 (49.0 mg, 0.324 mmol) and dichloromethane (5 mL) were added portionwise, followed by N-bromosuccinimide (63 mg, 0.356 mmol) and stirred at room temperature for 4 hours. After completion, the reaction was concentrated under reduced pressure at room temperature, and saturated aqueous sodium bicarbonate was slowly added to adjust the pH to 6-7. The mixture was extracted with ethyl acetate (10 mL x 4). The organic phases were combined and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using an eluent system (ethyl acetate:petroleum ether = 1:5) to afford 5-bromo-7-fluoro-3-methyl-dihydroindole 87 (61.0 mg, pale yellow solid) in an 82.0% yield. LCMS: m / z 229.8 / 231.8 (M+H).

[0451] Step 4. Synthesis of 1-(5-bromo-7-fluoro-3-methyl-indolin-1-yl)-2-(2-chloropyridin-3-yl)ethan-1-one (88)

[0452]

[0453] To a 50 mL flask were added compound 87 (61.0 mg, 0.266 mol), ethyl acetate (5 mL), T3P (50% ethyl acetate solution, w / w, 406.0 mg, 0.53 mmol), and diisopropylethylamine (103.0 mg, 0.80 mmol) in sequence at room temperature. The reaction was stirred for 3 hours. After completion, saturated aqueous sodium bicarbonate was slowly added to adjust the pH to 6-7. The mixture was extracted with ethyl acetate (10 mL x 4). The organic phases were combined, the solvent removed under reduced pressure, and the resulting residue was purified by silica gel column chromatography with an eluent system (ethyl acetate:petroleum ether = 1:3) to afford 1-(5-bromo-7-fluoro-3-methyl-indolin-1-yl)-2-(2-chloropyridin-3-yl)ethan-1-one 88 (79.0 mg, light yellow oil) in a 78.0% yield. LCMS: m / z 382.5 / 384.6 (M+H).

[0454] Step 5. Synthesis of 2-(2-chloropyridin-3-yl)-1-(7-fluoro-3-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-1-yl)ethan-1-one (89)

[0455]

[0456] In a dry 50 mL round-bottom flask, compound 88 (45.0 mg, 0.12 mmol), bis(pinacolato)diboron (36.0 mg, 0.14 mmol), potassium acetate (17.3 mg, 0.18 mmol), and (1,1'-bis(diphenylphosphino)ferrocene)palladium dichloride (4.8 mg, 0.0058 mmol) were added to 1,4-dioxane (10 ml) in sequence. The nitrogen atmosphere was replaced three times, the temperature was raised to 90°C, and the reaction was stirred for 3 hours. After completion of the reaction, the mixture was concentrated under reduced pressure and the resulting residue was purified by silica gel column chromatography with an eluent system (ethyl acetate:petroleum ether = 1:3) to afford 2-(2-chloropyridin-3-yl)-1-(7-fluoro-3-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-1-yl)ethan-1-one 89 (36.0 mg, light brown solid) in a 76.0% yield. LCMS: m / z 430.8 / 432.8 (M+H). Intermediate 57 : Synthesis of 2-(2-chloro-4-fluoro-pyridin-3-yl)acetic acid (57)

[0457]

[0458] Step 1. Synthesis of ethyl 2-(2-chloro-4-fluoropyridin-3-yl)acetate (56)

[0459]

[0460] At room temperature, compound 54 (1.0 g, 5.71 mmol), dichloromethane (25 mL), and N,N-dimethylformamide (1 mL) were added to a 50 mL round-bottom flask in sequence, and then a dichloromethane solution (10 mL) of oxalyl chloride (0.90 g, 7.14 mmol) was added dropwise. After the addition was complete, the reaction was stirred at room temperature for 1 hour, and the mixture was concentrated under reduced pressure. 15 mL of anhydrous tetrahydrofuran was added to the resulting residue, and compound 55 (2 M n-hexane solution, 5.2 mL, 10.3 mmol) and triethylamine (1.04 g, 10.3 mmol) in acetonitrile and tetrahydrofuran ( The mixture was added to a mixture of 20 mL:20 mL) and stirred at 0 ° C for 1 hour, then placed in a refrigerator for 16 hours, diluted with ethyl acetate (100 mL), washed with water, and the organic phase was adjusted to pH 4-5 with 0.5 mmol / L hydrochloric acid, stirred at room temperature for 5 minutes, and then adjusted to pH 8-9 with 1 mol / L sodium hydroxide aqueous solution. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was dissolved in 20 mL of ethanol, triethylamine (692 mg, 6.85 mmol) was added, and silver benzoate (silver benzoate) was added in batches at room temperature. The mixture was stirred for 10 minutes, heated to 80°C, stirred for 10 minutes, cooled to room temperature, filtered, and the filtrate concentrated. The residue was purified by column chromatography using an eluent system (ethyl acetate:petroleum ether = 1:7) to obtain ethyl 2-(2-chloro-4-fluoropyridin-3-yl)acetate 56 (250 mg, colorless oil) in a 20.0% yield. LCMS: m / z 217.8 / 219.8 (M+H).

[0461] Step 2. Synthesis of 2-(2-chloro-4-fluoro-pyridin-3-yl)acetic acid (57)

[0462]

[0463] Compound 56 (250 mg, 1.15 mmol) was dissolved in tetrahydrofuran (10 mL) at room temperature, and an aqueous solution of lithium hydroxide (10 mL, 0.5 mol / L) was added. The mixture was stirred at room temperature for 1 hour. After completion of the reaction, as monitored by LCMS, ethyl acetate (10 mL) was added, and the aqueous phase was adjusted to a pH of 3-4 with 0.5 mol / L dilute hydrochloric acid. The mixture was filtered, and the filter cake was washed with water and dried to yield 2-(2-chloro-4-fluoro-pyridin-3-yl)acetic acid 57 (220 mg, crude, pale yellow oil). LCMS: m / z 189.9 / 191.9 (M+H).

[0464] Similar to the preparation of intermediate 57, intermediates 65 and 83 were synthesized from the corresponding raw materials in the following table:

[0465]

[0466] Intermediate 82: Synthesis of 3-(2-chloropyridin-3-yl)propionic acid (82)

[0467]

[0468] Step 1. Synthesis of ethyl (E)-3-(2-chloropyridin-3-yl)acrylate (80)

[0469]

[0470] To a dry 100 mL round-bottom flask in an ice bath was added compound 79 (634 mg, 4.46 mmol), tetrahydrofuran (50 mL), and sodium hydride (357 mg, 8.93 mmol). The mixture was stirred in an ice bath for 0.5 hour, followed by the addition of compound 78 (1000 mg, 4.46 mmol). The mixture was gradually warmed to room temperature and stirred for 1 hour. The reaction system was monitored by LCMS, quenched with saturated aqueous ammonium chloride, extracted with ethyl acetate (50 mL x 3), and the organic phase was dried and concentrated. The resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to afford (E)-ethyl 3-(2-chloropyridin-3-yl)acrylate 80 (300 mg, white solid). Yield: 20%. LCMS: m / z 212.0 (M+H).

[0471] Step 2. Synthesis of ethyl 3-(2-chloropyridin-3-yl)propionate (81)

[0472]

[0473] To a dry 100 mL round-bottom flask at room temperature were added compound 80 (300 mg, 1.4 mmol), methanol (12 mL), water (3 mL), cuprous chloride (140 mg, 1.4 mmol), and sodium borohydride (54 mg, 1.4 mmol). The reaction was stirred at 0°C for 1 hour, followed by the addition of sodium borohydride (54 mg, 1.4 mmol). The reaction system was gradually warmed to room temperature and stirred for 1 hour. LCMS monitored the reaction completion. The reaction was quenched with ice water and extracted with ethyl acetate (50 mL x 3). The organic phase was dried and concentrated, and the resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to afford ethyl 3-(2-chloropyridin-3-yl)propanoate 81 (260 mg, white solid). Yield: 86%. LCMS: m / z 214.0 (M+H).

[0474] Step 3. Synthesis of 3-(2-chloropyridin-3-yl)propanoic acid (82)

[0475]

[0476] To a dry 100 mL round-bottom flask was added compound 81 (260 mg, 1.21 mmol), lithium hydroxide monohydrate (153 mg, 3.64 mmol), tetrahydrofuran (20 mL), methanol (4 mL), and water (4 mL). The mixture was stirred for 2 hours and monitored for completion by TLC. The reaction mixture was concentrated and lyophilized to afford 3-(2-chloropyridin-3-yl)propanoic acid 82 (300 mg as a white solid), which was used in the next step without purification. LCMS: m / z 185.9 (M+H).

[0477] Example:

[0478] Example P1:

[0479] Synthesis of Compound 2-(2-chloropyridin-3-yl)-1-(7-fluoro-5-(2-((1-hydroxypropyl-2-yl)amino)pyrimidin-4-yl)indolin-1-yl)ethan-1-one (P1)

[0480]

[0481] To a dry 100 mL round-bottom flask at room temperature, intermediate 13 (100 mg, 0.22 mmol), DL-aminopropanol (84 mg, 1.10 mmol), and DMSO (0.5 mL) were added. The temperature was raised to 90°C and stirred for 1 hour. After completion of the reaction, the mixture was concentrated under reduced pressure, 100 mL of water was added, and the mixture was extracted with ethyl acetate (150 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using ethyl acetate as the eluent to afford product P1 (10 mg, white solid) in a 10.0% yield. LCMS: m / z 441.9 (M+H).

[0482] 1 H-NMR (DMSO-d6,,400MHz): δ8.334(s,1H),8.333(d,J=10.4Hz,1H),7.91(s,1H),7. 76(m,3H),7.84(m,1H),7.83(d,J=10.4Hz,1H),7.43(dd,J=7.6Hz,5.2Hz,1H),7.14( d,J=5.2Hz,1H),6.85(d,J=8.0Hz,1H),4.71(t,J=5.6Hz,1H),4.29(t,J=8.0Hz,2H), 4.08(s,2H),3.50(m,1H),3.31(m,2H),3.23(t,J=8.4Hz,2H),1.16(d,J=6.4Hz,2H).

[0483] Similar to the preparation of Example P1, according to Figure 1 Examples P2-P7 were prepared from intermediate 13 and the corresponding amines according to the general synthetic scheme A given in:

[0484]

[0485] Embodiment P8:

[0486] Synthesis of 2-(2-chloropyridin-3-yl)-1-(7-fluoro-5-(2-((1-hydroxypropyl-2-yl)amino)pyridin-4-yl)indolin-1-yl)ethan-1-one (P8)

[0487]

[0488] Step 1. Synthesis of 2-((4-bromopyridin-2-yl)amino)propan-1-ol (24)

[0489]

[0490] To a dry 20 mL microwave reaction vial were added dimethyl sulfoxide (8 mL), 4-bromo-2-fluoropyridine (1 g, 0.0057 mol), 2-aminopropanol (0.65 g, 0.0085 mol), and N,N-diisopropylethylamine (1.1 g, 0.0085 mol). The temperature was raised to 140°C and the reaction was stirred for 0.5 hours. After completion of the reaction, as monitored by LCMS, the reaction solution was poured into water (10 mL) and extracted with ethyl acetate (10 mL × 3). The combined organic phases were washed with saturated brine (10 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography using an eluent system (ethyl acetate:petroleum ether = 1:1) to afford compound 24 (2-((4-bromopyridin-2-yl)amino)propan-1-ol) (0.8 g, yellow solid) in a 61% yield. LCMS: m / z 230.8 / 232.8 (M+H).

[0491] Step 2. Synthesis of 2-(2-chloropyridin-3-yl)-1-(7-fluoro-5-(2-((1-hydroxypropyl-2-yl)amino)pyridin-4-yl)indolin-1-yl)ethan-1-one (P8)

[0492]

[0493] To a dry 100 mL round-bottom flask at room temperature were added Intermediate 24 (0.05 g, 0.00021 mol), K2CO3 (0.058 g, 0.00042 mol), Intermediate 11 (0.108 g, 0.00026 mol), Pd(dppf)Cl2 (15 mg, 0.000021 mol), 1,4-dioxane (20 mL), and distilled water (2 mL). The atmosphere was purged with nitrogen three times. The temperature was raised to 80°C under nitrogen and stirred for 5 hours. After completion of the reaction, the mixture was concentrated under reduced pressure, and the crude product was purified by thin-layer chromatography (ethyl acetate) to afford compound P8 (0.04 g, white solid) in a 43% yield. LCMS: m / z 440.7 (M+H).

[0494] 1 H-NMR (DMSO-d6, 400MHz): δ8.34(d,J=3.6Hz,1H),7.98(d,J=5.8Hz,1H),7.89(d,J=6.8Hz,1H),7.51(s,1H),7.45-7.40(m,2H),6.96(br s,1H),6.88(br s,1H),4.91(br s,1H),4.28(t,J=7.8Hz,2H),4.08(s,2H),4.05-3.96(m,1H),3.52-3.44(m,1H),3.37(s,1H),3.22(t,J=7.8Hz,2H),1.15(d,J=6.5Hz,3H).

[0495] Similar to the preparation of Example P8, according to Figure 2 Examples P10-P16, P32, P34, P36, P44, P47-51, P53 and P58-59 were synthesized using intermediates 11, 53 or borate intermediates similar to intermediate 11 according to the general synthetic scheme B given in

[0496]

[0497]

[0498]

[0499]

[0500]

[0501]

[0502] Embodiment P9:

[0503] Synthesis of 2-(2-chloropyridin-3-yl)-1-(7-fluoro-5-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)indolin-1-yl)ethan-1-one (P9)

[0504]

[0505] Step 1. Synthesis of tert-butyl (4-(7-fluoroindolin-5-yl)pyridin-2-yl)(1-methyl-1H-pyrazol-5-yl)carbamate (25)

[0506]

[0507] Method 1: To a dry 50 mL round-bottom flask at room temperature, intermediate 4 (600 mg, 2.279 mmol), 1,4-dioxane (4 mL), water (0.8 mL), intermediate 8 (563 mg, 1.595 mmol), Pd(dppf)Cl2 (167 mg, 0.2279 mmol), and sodium bicarbonate (383 mg, 4.56 mmol) were added. The atmosphere was purged with nitrogen three times. The mixture was heated to 75°C and stirred for 2 hours. After completion of the reaction, the mixture was filtered while hot and evaporated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography using an eluent system (ethyl acetate / petroleum ether = 1 / 5 to 3 / 1) to obtain tert-butyl intermediate 25 (4-(7-fluoroindolin-5-yl)pyridin-2-yl)(1-methyl-1H-pyrazol-5-yl)carbamate (620 mg, solid) in a yield of 60.45%. 1 H NMR (400MHz, DMSO) δ8.27(d,J=5.2Hz,1H),7.85(d,J=1.2Hz,1H),7.49(dd,J=5.2,1.6Hz,1H),7.43(s,1H),7.41–7.36(m, 2H),6.18(d,J=2.0Hz,1H),6.14(s,1H),3.70(s,3H),3.57(t,J=8.8Hz,2H),3.08(t,J=8.8Hz,2H),1.41(s,9H).LCMS:m / z 410.2(M+H).

[0508] Method 2: To a dry 250 mL round-bottom flask at room temperature, intermediate 8 (0.984 g, 2.80 mmol), intermediate 4 (0.88 g, 2.90 mol), triethylamine (1.19 g, 12 mmol), 1,4-dioxane (15 mL), and distilled water (3 mL) were added sequentially. Pd(dppf)Cl2 (240 mg, 0.3 mol) was then added. The nitrogen atmosphere was replaced three times, and the temperature was raised to 70°C. The reaction was stirred for 16 hours. After completion of the reaction, as monitored by LCMS, the product was concentrated under reduced pressure and the residue was purified by silica gel column chromatography using an eluent system (ethyl acetate:petroleum ether = 1:3 to 1:5) to afford intermediate 25 (tert-butyl 4-(7-fluoroindolin-5-yl)pyridin-2-yl)(1-methyl-1H-pyrazol-5-yl)carbamate) (1.0 g, solid, 88% yield). LCMS: m / z 410.1 (M+H).

[0509] Method 3: To a round-bottom flask (50 mL), add Intermediate 8 (1.80 g), Intermediate 4 (2.15 g), methyltetrahydrofuran (15 mL), triethylamine (2.06 g), purified water (3.6 g), and Pd(ddpf)Cl2 (1.25 g). Replace the atmosphere with nitrogen three times, maintain the temperature at 65-75°C, and react for approximately 15 hours. After completion of the reaction, sample the mixture for HPLC analysis and concentrate under reduced pressure. Add MTBE and tap water, stir to dissolve, filter through diatomaceous earth, and wash the filter cake. Combine the filtrates, separate the layers, and wash the organic layer twice with tap water. After concentrating the combined filtrates under reduced pressure, add n-heptane dropwise, cool, and slurry. Filter under reduced pressure, and rinse the filter cake with methyl tert-butyl ether. The filter cake was dried under reduced pressure to obtain 1.72 g of Intermediate 25 (tert-butyl 4-(7-fluoroindolin-5-yl)pyridin-2-yl)(1-methyl-1H-pyrazol-5-yl)carbamate). LCMS: m / z 410.1 (M+H).

[0510] Step 2. Synthesis of (4-(1-(2-(2-chloropyridin-3-yl)acetyl)-7-fluoroindolin-5-yl)pyridin-2-yl)(1-methyl-1H-pyrazol-5-yl)carbamate (26)

[0511]

[0512] Method 1: To a dry 25 mL round-bottom flask at room temperature, compound 9 (2-chloropyridine-3-acetic acid) (493 mg, 2.876 mmol), AcO (285 mg, 2.80 mmol), and THF (3 mL) were added. The reaction was incubated at 75°C for 1 hour. Intermediate 25 (620 mg, 1.514 mmol), THF (2 mL), DMF (1 mL), and pyridine (240 mg, 3.028 mmol) were then added. The atmosphere was purged with nitrogen three times. The temperature was raised to 70°C and stirred for 3 hours. After completion of the reaction, methanol (2 mL) was added, the mixture was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography using an eluent system (ethyl acetate / petroleum ether = 1 / 4 to DCM:MeOH = 30 / 1) to afford Intermediate 26 (400 mg, light yellow solid) in a 46.9% yield. LCMS: m / z 564.2 (M+H).

[0513] Method 2: Compound 25 (1.682 g, 4.1 mmol), diisopropylethylamine (2.121 g, 16.4 mmol), compound 9 (0.843 g, 4.9 mol), ethyl acetate (20 mL), and 1-propylphosphonic anhydride (50% ethyl acetate solution, 6.54 g, 10 mmol) were added to a 1000 mL round-bottom flask at room temperature. The mixture was stirred at room temperature for 5 hours. After completion of the reaction, saturated sodium bicarbonate solution was added to adjust the pH to 7-8, and the mixture was extracted with ethyl acetate (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain tert-butyl (4-(1-(2-(2-chloropyridin-3-yl)acetyl)-7-fluoroindolin-5-yl)pyridin-2-yl)(1-methyl-1-hydrogen-pyrazol-5-yl)carbamate (26) (2.033 g, pale yellow oil, crude product). LCMS: m / z 562.5 / 564.5 (M+H).

[0514] Step 3. Synthesis of 2-(2-chloropyridin-3-yl)-1-(7-fluoro-5-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)indolin-1-yl)ethan-1-one (P9)

[0515]

[0516] Method 1: To a dry 25 mL single-necked vial, intermediate 26 (350 mg, 0.622 mmol), TFA (709 mg, 6.22 mmol), acetonitrile (5 mL), and H₂O (0.5 mL) were added at room temperature and stirred at 40°C under nitrogen for 8 hours. After completion of the reaction, as determined by LCMS, the crude product was purified by preparative liquid chromatography to afford product P9 (2-(2-chloropyridin-3-yl)-1-(7-fluoro-5-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)indolin-1-yl)ethan-1-one) (135 mg, yellow solid) in a 46.9% yield. LCMS: m / z 463.1 (M+H).

[0517] 1 H-NMR(400MHz,DMSO-d6)δ8.82(s,1H),8.35(dd,J=4.8,1.6Hz,1H),8.17(d, J=5.6Hz,1H),7.88(dd,J=7.2,1.2Hz,1H),7.50(s,1H),7.45–7.41(m,2H),7 .34(d,J=1.2Hz,1H),7.10(d,J=5.2Hz,1H),7.02(s,1H),6.28(d,J=1.2Hz,1 H),4.28(t,J=8.0Hz,2H),4.08(s,2H),3.69(s,3H),3.21(t,J=8.0Hz,2H)..

[0518] Method 2: To a dry 50 mL round-bottom flask, dichloromethane (15 mL) and intermediate 26 (2.033 g, 3.6 mmol) were added sequentially, followed by the slow addition of trifluoroacetic acid (4 mL). The reaction was stirred at room temperature overnight. After completion of the reaction, saturated aqueous sodium carbonate solution was added dropwise to adjust the pH to 7-8, monitored by LCMS. The mixture was separated and extracted with dichloromethane (50 mL x 3). The combined organic phases were concentrated under reduced pressure, and water (20 mL) was added dropwise to the suspension with stirring. The mixture was stirred in an ice bath for 1-2 hours. The mixture was filtered, and the filter cake was washed once with water (10 mL). The filter cake was then dried under reduced pressure to yield 1.63 g of P9 (2-(2-chloropyridin-3-yl)-1-(7-fluoro-5-(2-((1-methyl-1-hydrogen-pyrazol-5-yl)amino)pyridin-4-yl)indolin-1-yl)ethan-1-one). LCMS: m / z 463.1 (M+H).

[0519] Similar to the preparation of Example P9, according to Figure 3 Examples P45-46 and P61 were synthesized using intermediate 25 or an intermediate similar thereto according to the general synthetic scheme C given in:

[0520]

[0521] Embodiment P17:

[0522] Synthesis of 1-(7-fluoro-5-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyrimidin-4-yl)indolin-1-yl)-2-(2-fluorophenyl)ethan-1-one (P17)

[0523]

[0524] Step 1. Synthesis of N-4-(7-fluoroindolin-5-yl)-N-(1-methyl-1H-pyrazol-5-yl)pyrimidin-2-ylamine (32)

[0525]

[0526] To a dry 100 mL round-bottom flask were added compound 4 7-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indoline (900 mg, 3.80 mmol), compound 17 4-chloro-N-(1-methyl-1H-pyrazol-5-yl)pyrimidin-2-amine (800 mg, 3.80 mmol), potassium carbonate (1.1 g, 7.60 mmol), (1,1-bis(diphenylphosphino)ferrocene)palladium dichloride (278 mg, 0.38 mmol) and 1,4-dioxane (20.0 mL) at room temperature. The atmosphere was replaced with nitrogen five times, the temperature was raised to 80°C, and the reaction was stirred overnight. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, 100 mL of water was added, and the mixture was extracted with ethyl acetate (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with an eluent system (ethyl acetate:petroleum ether = 1:10) to give intermediate 32N-4-(7-fluoroindolin-5-yl)-N-(1-methyl-1H-pyrazol-5-yl)pyrimidin-2-ylamine (150 mg, white solid) in a yield of 12.0%.

[0527] LCMS: m / z 311.0 (M+H).

[0528] Step 2. Synthesis of 1-(7-fluoro-5-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyrimidin-4-yl)indolin-1-yl)-2-(2-fluorophenyl)ethan-1-one (P17)

[0529]

[0530] Compound 32N-4-(7-fluoroindolin-5-yl)-N-(1-methyl-1H-pyrazol-5-yl)pyrimidin-2-ylamine (50 mg, 0.16 mmol), compound 33 2-(2-Fluorophenyl)acetic acid (25 mg, 0.16 mmol), N,N-diisopropylethylamine (0.1 mL, 0.64 mmol), 1-propylphosphonic anhydride (407 mg, 50% (wt%) ethyl acetate solution (0.64 mmol) and N,N-dimethylformamide (1.0 mL) were stirred and reacted for 30 minutes. After completion of the reaction, the mixture was concentrated under reduced pressure, 100 mL of water was added, and the mixture was extracted with ethyl acetate (150 mL × 3). The organic phases were combined, washed with saturated brine (50 mL × 5), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with an eluent system (ethyl acetate: petroleum ether = 1:1) to obtain the product P17. 1-(7-Fluoro-5-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyrimidin-4-yl)indolin-1-yl)-2-(2-fluorophenyl)ethan-1-one (16.7 mg, pale yellow solid), yield: 20.0%. LCMS: m / z 447.0 (M+H).

[0531] 1 H NMR (DMSO-d6, 400MHz): δ9.48(s,1H),8.51(d,J=5.2Hz,1H),7.91(s,1H),7.83(d,J=12.4Hz,1H),7.46(d,J=5.2Hz,1H),7. 41-7.31(m,3H),7.19(q,J=7.6Hz,2H),6.29(s,1H),4.26(t,J=8.0Hz,2H),3.98(s,2H),3.70(s,3H),3.23(t,J=8.0Hz,2H).

[0532] according to Figure 3 The following examples P18-P20, P23-P25, P28-31, P33, P37-39 and P54-57 were synthesized using intermediate 32 or an intermediate similar thereto according to the general synthetic scheme C given in:

[0533]

[0534]

[0535]

[0536]

[0537]

[0538] Embodiment P35:

[0539]

[0540] Step 1. Synthesis of methyl 1-methyl-5-nitro-1H-pyrazole-3-carboxylate (59)

[0541]

[0542] To a 100 mL round-bottom flask in an ice-water bath, compound 58 (5.0 g, 12.74 mmol), potassium carbonate (1.94 g, 14.0 mmol), and N,N-dimethylformamide (50 mL) were added in sequence, and a solution of iodomethane (3.80 g, 26.75 mmol) in N,N-dimethylformamide (10 mL) was slowly added dropwise. After the addition was complete, the reaction was stirred for 16 hours. After completion of the reaction, water (150 mL) was added, and the mixture was extracted with ethyl acetate (50 mL × 3). The organic phases were combined, washed with saturated brine (30 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography with an eluent system (ethyl acetate:petroleum ether = 1:20) to give methyl 1-methyl-5-nitro-1H-pyrazole-3-carboxylate 59 (1.0 g, white solid) in a yield of 17.0%. LCMS: m / z 185.9 (M+H)

[0543] Step 2. Synthesis of methyl 5-amino-1-methyl-1H-pyrazole-3-carboxylate (60)

[0544]

[0545] Compound 59 (200 mg, 1.081 mmol) was dissolved in methanol (10 mL) at room temperature, and Pd / C (10%, 90 mg) was added. The mixture was stirred under 1 atm of hydrogen for 4 hours. After completion of the reaction, the filtrate was filtered and concentrated under reduced pressure to afford methyl 5-amino-1-methyl-1H-pyrazole-3-carboxylate 60 (145 mg, light yellow oil). Yield: 87.0%. LCMS: m / z 156 (M+H).

[0546] Step 3. Synthesis of methyl 5-((4-bromopyridin-2-yl)amino)-1-methyl-1H-pyrazole-3-carboxylate (61)

[0547]

[0548] To a 50 mL round-bottom flask at room temperature, compound 60 (650 mg, 4.19 mmol) and N,N-dimethylformamide (10 mL) were added sequentially. Sodium hydride (335 mg, 8.38 mmol) was added portionwise and the mixture was stirred for 0.5 h. 2-Bromo-4-fluoropyridine (1.47 g, 8.387 mmol) was then added and the mixture was stirred for 16 h. After completion of the reaction, monitored by LCMS, water (30 mL) was added and the mixture was extracted with ethyl acetate (10 mL × 5). The combined organic phases were washed with saturated brine (20 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography using an eluent system (ethyl acetate:petroleum ether = 1:4) to afford methyl 5-((4-bromopyridin-2-yl)amino)-1-methyl-1H-pyrazole-3-carboxylate 61 (130 mg, light yellow solid) in a 10% yield. LCMS: m / z 310.9 / 312.9 (M+H).

[0549] Step 4. Synthesis of (5-((4-bromopyridin-2-yl)amino)-1-methyl-1H-pyrazol-3-yl)methanol (62)

[0550]

[0551] Compound 61 (130 mg, 0.418 mmol) was dissolved in anhydrous methanol (25 mL) at room temperature, and sodium borohydride (156 mg, 4.18 mmol) was added portionwise. After addition, the reaction was stirred at room temperature for 2 hours and concentrated under reduced pressure. The resulting residue was purified by column chromatography with an eluent system (ethyl acetate:petroleum ether = 4:1) to provide (5-((4-bromopyridin-2-yl)amino)-1-methyl-1H-pyrazol-3-yl)methanol 62 (100 mg, white gum) in an 85% yield. LCMS: m / z 282.9 / 284.9 (M+H).

[0552] Step 5. Synthesis of N-(3-(((tert-Butyldimethylsilyl)oxy)methyl)-1-methyl-1H-pyrazol-5-yl)-N-(4-bromopyridin-2-yl)-amine (63)

[0553]

[0554] To a 50 mL round-bottom flask at room temperature, compound 62 (100 mg, 0.355 mmol), tert-butyldimethylsilyl chloride (532 mg, 3.55 mmol), triethylamine (358 mg, 3.55 mmol), 4-dimethylaminopyridine (4.33 mg, 0.036 mmol) and dichloromethane (10 mL) were added in sequence, and the nitrogen atmosphere was replaced three times. The reaction was stirred at room temperature for 16 hours. After completion of the reaction, water (30 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with an eluent system (ethyl acetate:petroleum ether = 1:5) to afford N-(3-(((tert-butyldimethylsilyl)oxy)methyl)-1-methyl-1H-pyrazol-5-yl)-N-(4-bromopyridin-2-yl)-amine 63 (50 mg, light yellow oil) in a 33.0% yield. LCMS: m / z 396.9 / 398.9 (M+H).

[0555] Step 6. Synthesis of 1-(5-(2-((3-((tert-butyldimethylsilyl)oxy)methyl)-1-methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)-7-fluoroindolin-1-yl)-2-(2-chloropyridin-3-yl)ethan-1-one (64)

[0556]

[0557] Into a 50 mL round-bottom flask were added compound 63 (50 mg, 0.126 mmol), 1,4-dioxane (10 mL), PdCl2(dppf) (9.2 mg, 0.0126 mmol), potassium carbonate (26.1 mg, 0.189 mmol), and compound 11 (52.5 mg, 0.126 mmol) in sequence at room temperature. The nitrogen atmosphere was replaced three times, the temperature was raised to 80°C, and the reaction was stirred for 3 hours. After completion of the reaction, the mixture was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with an eluent system (ethyl acetate:petroleum ether = 5:1) to afford 1-(5-(2-((3-(((tert-butyldimethylsilyl)oxy)methyl)-1-methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)-7-fluoroindolin-1-yl)-2-(2-chloropyridin-3-yl)ethan-1-one 64 (60 mg, light yellow oil) in a 78.5% yield. LCMS: m / z 606.5 / 608.5, (M+H)

[0558] Step 7. Synthesis of 2-(2-chloropyridin-3-yl)-1-(7-fluoro-5-(2-((3-(hydroxymethyl)-1-methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)indolin-1-yl)ethan-1-one (P35)

[0559]

[0560] Compound 64 (60 mg, 0.099 mmol), trifluoroacetic acid (2 mL), and dichloromethane (10 mL) were added sequentially to a 50 mL flask at room temperature and stirred for 1 hour. After completion of the reaction, the mixture was concentrated under reduced pressure, and saturated aqueous sodium bicarbonate (15 mL) was slowly added. The mixture was extracted with ethyl acetate (10 mL x 5). The organic phases were combined and concentrated under reduced pressure. The residue was purified by preparative liquid chromatography (column: Gemini-C18 150 x 21.2 mm, 5 μm; mobile phase: ACN-H2O (0.1% FA), gradient: 10-60) to afford 2-(2-chloropyridin-3-yl)-1-(7-fluoro-5-(2-((3-(hydroxymethyl)-1-methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)indolin-1-yl)ethan-1-one P35 (12 mg, light yellow solid) in a 25% yield.

[0561] 1 H-NMR (CD3OD, 400MHz): 8.33 (d, J = 3.2Hz, 1H), 8.15 (d, J = 5.6Hz, 1H), 7.90 (d, J = 7.6Hz, 1H), 7.51 (s, 1H), 7.44-7.38 (m, 2H), 7.12 (d, J = 5.6Hz, 1 H),6.99(s,1H),6.31(s,1H),4.56(s,2H),4.37(t,J=7.6Hz,2H),4.14(s,2H),3.73(s,3H),3.28(t,J=7.2Hz,2H).LCMS:m / z492.9 / 494.9(M+H)

[0562] Embodiment P60: Synthesis of 2-(2-chloropyridin-3-yl)-1-(7-fluoro-5-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)indolin-1-yl)-2-hydroxyethan-1-one (P60)

[0563]

[0564] Step 1. Synthesis of ethyl 2-(2-chloropyridin-3-yl)-2-hydroxyacetate (91)

[0565]

[0566] To a dry 250 mL round-bottom flask in an ice bath, compound 90 (6.0 g, 0.031 mol), tetrahydrofuran (100 mL), and isopropyllithium chloride and magnesium chloride (31 mL, 40.3 mol) were added. The atmosphere was purged with nitrogen three times, and the mixture was allowed to warm to room temperature and stirred for 3 hours. The mixture was cooled in an ice-water bath, and compound 2 (3.2 g, 0.031 mol) was added. The mixture was gradually warmed to room temperature and stirred for 2 hours. The reaction system was quenched with saturated ammonium chloride (100 mL) and extracted with ethyl acetate (80 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 1:10) to afford ethyl 2-(2-chloropyridin-3-yl)-2-hydroxyacetate 91 (1.4 g, light yellow oil) in a 21% yield. LCMS: m / z 216.0 (M+H).

[0567] Step 2. Synthesis of ethyl 2-((tert-butyldimethylsilyl)oxy)-2-(2-chloropyridin-3-yl)acetate (92)

[0568]

[0569] To a dry 100 mL round-bottom flask at room temperature were added compound 91 (800 mg, 3.7 mmol), dichloromethane (50 mL), tert-butyldimethylsilyl chloride (2796 mg, 18.5 mmol), 4-dimethylaminopyridine (452 ​​mg, 3.7 mmol), and triethylamine (3740 mg, 37.0 mmol). The atmosphere was purged with nitrogen three times and the mixture was stirred at room temperature for 18 hours. After completion, the reaction was quenched with ice water (100 mL) and extracted with dichloromethane (60 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 1:20) to afford ethyl 2-((tert-butyldimethylsilyl)oxy)-2-(2-chloropyridin-3-yl)acetate 92 (640 mg, colorless oil) in a 52% yield. LCMS: m / z 330.0 (M+H).

[0570] Step 3. Synthesis of 2-((tert-butyldimethylsilyl)oxy)-2-(2-chloropyridin-3-yl)acetic acid (93)

[0571]

[0572] Compound 92 (640 mg, 1.94 mmol), lithium hydroxide monohydrate (244 mg, 5.81 mmol), tetrahydrofuran (20 mL), and water (4 mL) were added to a dry 100 mL round-bottom flask at room temperature and stirred for 2 hours. The mixture was filtered, concentrated under reduced pressure, and the pH was adjusted to 7 with 1N aqueous hydrochloric acid. A solid precipitated, which was filtered and dried to afford 2-((tert-butyldimethylsilyl)oxy)-2-(2-chloropyridin-3-yl)acetic acid 93 (180 mg, white solid) in a 31% yield. LCMS: m / z 302.0 (M+H). Step 4. Synthesis of tert-butyl N-(4-(1-(2-((tert-butyldimethylsilyl)oxy)-2-(2-chloropyridin-3-yl)acetyl)-7-fluoroindol-5-yl)pyridin-2-yl)-N-(1-methyl-1H-pyrazol-5-yl)carbamate (94)

[0573]

[0574] To a dry 100 mL round-bottom flask in an ice bath were added compound 93 (243 mg, 0.6 mmol), compound 25 (180 mg, 0.6 mmol), phosphorus oxychloride (273 mg, 1.8 mmol), and pyridine (20 mL). The mixture was gradually warmed to room temperature and stirred for 3 hours. Iced water (50 mL) was added for dilution and extraction with ethyl acetate (50 mL x 3). The combined organic phases were concentrated under reduced pressure, and the resulting residue was purified by preparative TLC (ethyl acetate:petroleum ether = 1:3) to afford tert-butyl N-(4-(1-(2-((tert-butyldimethylsilyl)oxy)-2-(2-chloropyridin-3-yl)acetyl)-7-fluoroindol-5-yl)pyridin-2-yl)-N-(1-methyl-1H-pyrazol-5-yl)carbamate 94 (250 mg) in a 61% yield. LCMS: m / z 692.5 (M+H).

[0575] Step 5. Synthesis of 2-((tert-butyldimethylsilyl)oxy)-2-(2-chloropyridin-3-yl)-1-(7-fluoro-5-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)indolin-1-yl)ethan-1-one (95)

[0576]

[0577] Compound 94 (250 mg, 0.36 mmol), dichloromethane (20 mL), and trifluoroacetic acid (5 mL) were added to a dry 100 mL round-bottom flask at room temperature and stirred for 2 hours. The mixture was cooled in an ice bath, neutralized with a saturated aqueous sodium bicarbonate solution (50 mL), and extracted with dichloromethane (30 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to afford 2-((tert-butyldimethylsilyl)oxy)-2-(2-chloropyridin-3-yl)-1-(7-fluoro-5-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)indolin-1-yl)ethan-1-one 95 (200 mg, colorless oil) in a 93% yield. LCMS: m / z 593.1 (M+H).

[0578] Step 6. Synthesis of 2-(2-chloropyridin-3-yl)-1-(7-fluoro-5-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)indolin-1-yl)-2-hydroxyethan-1-one (P60)

[0579]

[0580] Compound 95 (200 mg, 0.34 mmol), tetrahydrofuran (4 mL), and tetrabutylammonium fluoride (1N THF solution, 4 mL) were added to a dry 100 mL round-bottom flask at room temperature, and the mixture was stirred at room temperature for 1 hour. The mixture was diluted with water (50 mL), extracted with ethyl acetate (30 mL×3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether=1:20) to give 2-(2-chloropyridin-3-yl)-1-(7-fluoro-5-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)dihydroindole-1-yl)-2-hydroxyethan-1-one P60 (120 mg, white solid), which was recrystallized from ethyl acetate:n-hexane=1:2 to give 2-(2-chloropyridin-3-yl)-1-(7-fluoro-5-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)dihydroindole-1-yl)-2-hydroxyethan-1-one P60 (55 mg, white solid) in a yield of 34%.

[0581] 1H NMR (400MHz, DMSO-d6) δ8.82 (s, 1H), 8.38 (dd, J = 2.0, 4.8Hz, 1H), 8.16 (d, J = 5.6Hz, 1H), 8 .02(dd,J=2.0,7.6Hz,1H),7.55-7.47(m,2H),7.41(d,J=12Hz,1H),7.34(d,J=1.6Hz,1H), 7.09(dd,J=1.2,5.2Hz,1H),7.01(s,1H),6.63(d,J=6.8Hz,1H),6.27(d,J=2.0Hz,1H),5. 77(d,J=6.4Hz,1H),4.42-4.33(m,1H),4.24-4.14(m,1H),3.68(s,3H),3.27-3.20(m,1H).

[0582] LCMS: m / z 479.0 (M+H).

[0583] Similar to the preparation method of Example P60, Example P64 was synthesized using an intermediate similar to Intermediate 25:

[0584]

[0585] Step 7: Compound P60 was resolved to produce compounds P62 and P63

[0586]

[0587] Split conditions:

[0588] Chiral column: AD-H, 0.46cm ID x 15cm L

[0589] Mobile phase: HEP:IPA (0.1% DEA) = 60:40

[0590] Flow rate: 0.5 mL

[0591] Detection wavelength: UV 254nm

[0592] Column temperature: 25°C

[0593] The first peak compound (Peak1) is numbered P62

[0594] 1H NMR (400MHz, DMSO) δ8.81(s,1H),8.38(dd,J=4.8,1.8Hz,1H),8.16(d,J=5.4Hz,1H),8.02(dd, J=7.6,1.6Hz,1H),7.52(dd,J=7.6,4.8Hz,1H),7.48(s,1H),7.40(d,J=12.0Hz,1H),7.34(d,J =1.8Hz,1H),7.09(d,J=4.2Hz,1H),7.00(s,1H),6.61(s,1H),6.27(d,J=1.6Hz,1H),5.77(d,J =6.6Hz,1H),4.37-4.41(m,1H),4.26–4.14(m,1H),3.68(s,3H),3.22(dd,J=13.4,7.2Hz,,2H).

[0595] .LCMS: m / z 479.0 (M+H).

[0596] The second peak compound (Peak2) is numbered P63

[0597] 1 H NMR (400MHz, DMSO) δ8.81(s,1H),8.38(dd,J=4.8,1.8Hz,1H),8.16(d,J=5.4Hz,1H),8.02(dd,J=7 .6,1.8Hz,1H),7.52(dd,J=7.6,4.8Hz,1H),7.48(s,1H),7.40(d,J=12.0Hz,1H),7.34(d,J=1.8Hz ,1H),7.09(dd,J=5.4,1.6Hz,1H),7.00(s,1H),6.61(d,J=6.8Hz,1H),6.27(d,J=1.8Hz,1H),5.77 (d,J=6.8Hz,1H),4.43–4.35(m,1H),4.25-4.15(m,1H),3.68(s,3H),3.22(dd,J=13.4,7.2Hz,2H).

[0598] LCMS: m / z 479.0 (M+H).

[0599] Examples P40 and P41. Resolution of compound P53 to prepare P40 and P41

[0600]

[0601] Split conditions:

[0602] Chiral column: Chiralpak-OJ, 0.46 cm ID × 25 cm L

[0603] Mobile phase: HEX-EtOH (0.2% DEA) = 50:50

[0604] Flow rate: 0.8 mL

[0605] Detection wavelength: UV 214 / 254nm

[0606] Column temperature: 40°C

[0607] The first peak compound (Peak1) is numbered P40

[0608] 1 H NMR (CD3OD, 400MHz): 8.34 (d, J = 4.8Hz, 1H), 7.98 (d, J = 5.6Hz, 1H), 7.90 (d, J = 7.6Hz, 1H), 7.51 (s, 1H), 7.44-7.38 (m, 2H), 6.94-6.92 (m, 1H), 6. 79(s,1H),4.51-4.48(m,1H),4.37(t,J=8Hz,2H),4.30-4.27(m,1H),4. 14(s,2H),3.28(t,J=7.6Hz,2H),2.41-2.38(m,2H),2.35-2.31(m,2H).

[0609] LCMS: m / z 452.8 / 454.9 (M+H)

[0610] The second peak compound (Peak2) is numbered P41

[0611] 1 H NMR(CD3OD,400MHz):8.33(d,J=4.8Hz,1H),7.97(d,J=6Hz,1H),7.90(d,J=7.6Hz,1H),7.53(s,1H),7.43-7.40(m,2H),6.96(d,J=5.6Hz,1H),6 .84(s,1H),4.37(t,J=7.6Hz,2H),4.14(s,2H),4.10-4.03(m,1H),3.85 -3.77(m,1H),3.29(t,J=8Hz,2H),2.91-2.85(m,2H),1.92-1.85(m,2H).

[0612] LCMS: m / z 452.8 / 454.9 (M+H)

[0613] Examples P42 and P43. Resolution of Compound P16 to Prepare P42 and P43

[0614]

[0615] Split conditions:

[0616] Chiral column: OJ, 0.46cm ID*25cm L

[0617] Mobile phase: n-hexane: ethanol (0.2% diethylamine) = 50:50

[0618] Flow rate: 0.8 mL

[0619] Detection wavelength: UV 214 / 254nm

[0620] Column temperature: 40°C

[0621] The first peak compound (Peak1) is numbered P42

[0622] 1 H NMR (400MHz, DMSO-d6) δ8.82(s,1H),8.34(d,J=3.2Hz,1H),8.16(d,J=5.2Hz,1H),7.89(d, J=7.6Hz,1H),7.52(s,1H),7.49-7.41(m,2H),7.34(d,J=1.4Hz,1H),7.10(d,J=5.4Hz,1H) ,7.01(s,1H),6.28(s,1H),4.97-4.89(m,1H),4.18(d,J=16.8Hz,1H),3.96(d,J=16.8Hz,1 H),3.68(s,3H),3.57-3.51(m,1H),2.72(d,J=16.0Hz,1H),1.27(d,J=6.4Hz,3H).LCMS:m / z 476.9(M+H).

[0623] The second peak compound (Peak2) is numbered P43

[0624] 1H NMR (400MHz, DMSO-d6) δ8.82(s,1H),8.34(d,J=3.6Hz,1H),8.16(d,J=5.4Hz,1H),7.8 9(d,J=6.8Hz,1H),7.52(s,1H),7.48-7.41(m,2H),7.34(s,1H),7.10(d,J=5.4Hz,1H) ,7.01(s,1H),6.27(s,1H),4.98-4.89(m,1H),4.18(d,J=16.8Hz,1H),3.96(d,J=16.8 Hz,1H),3.68(s,3H),3.57-3.51(m,1H),2.72(d,J=16.0Hz,1H),1.27(d,J=6.4Hz,3H).

[0625] LCMS: m / z 476.9 (M+H).

[0626] Effect Example I: Chemical Stability Test

[0627] 1. Means and conditions used for testing in chemical stability tests

[0628] Detection method: Ultra-performance liquid chromatography (UPLC)

[0629] Chromatographic conditions:

[0630] System: Ultra-high performance liquid chromatography system, including pump, autosampler, detector and column oven

[0631] Column: Waters Acquity UPLC BEH C18 (2.1*50mm, 1.7μm)

[0632] Detector: PDA detector

[0633] Detection wavelength: 225nm

[0634] Mobile phase: A: 0.05% trifluoroacetic acid in water

[0635] B: Acetonitrile

[0636] gradient:

[0637] Flow rate: 0.4ml / min

[0638] Column temperature: 40°C

[0639] Injection volume: 2μl

[0640] 2. Investigation of the chemical stability of the compounds of the present invention

[0641] (1) Preparation of solution of test compound P9

[0642] Compound P9 was prepared into 0.2 mg / ml solutions in different buffer systems using PEG400 as a solubilizer for chemical stability studies. Solutions of different pH values ​​were prepared as follows:

[0643]

[0644] Chemical stability test conditions: Solutions of compound P9 with different pH values ​​were placed at 37° C. for 24 hours, and the content of compound P9 was determined by HPLC at 0, 4, 8, 12, 14 and 24 hours, respectively.

[0645] (2) Chemical stability test results of the test compound P9 in solutions with different pH values

[0646]

[0647] (3) Conclusion

[0648] The research results show that compound P9 has good chemical stability and no obvious impurity growth after being placed at 37°C for 24 hours in solutions with pH 2.0, pH 6.8, and pH 7.4.

[0649] 3. Chemical stability study of comparative compounds

[0650] Similarly, the chemical stability of compound A107 in WO2017 / 114510A1 in solutions at pH 1.2, pH 6.8, and pH 7.4 was tested using the above method.

[0651] (1) Preparation of compound A107 solution

[0652] (1.1) A107 Prescription Solution (pH 1.2):

[0653] Solution prescription: 10% PEG400 + 5% Solutol HS-15 + 85% pH1.2 dilute hydrochloric acid

[0654] Solution concentration: 0.2mg / ml

[0655] Preparation method: Weigh A107, add the prescribed amount of PEG400 and Solutol HS-15, vortex to obtain a clear solution, then add the prescribed amount of pH 1.2 dilute hydrochloric acid and mix well.

[0656] (1.2) A107 prescription solution (pH 6.8):

[0657] Solution prescription: 10% PEG400 + 5% Solutol HS-15 + 85% pH 6.8 phosphate buffer

[0658] Solution concentration: 0.2mg / ml

[0659] Preparation method: Weigh A107, add the prescribed amount of PEG400 and Solutol HS-15, vortex to obtain a clear solution, then add the prescribed amount of pH 6.8 phosphate buffer and mix well.

[0660] (1.3) A107 Prescription Solution (pH 7.4):

[0661] Solution prescription: 10% PEG400 + 5% Solutol HS-15 + 85% pH 7.4 phosphate buffer

[0662] Solution concentration: 0.2mg / ml

[0663] Preparation method: Weigh A107, add the prescribed amount of PEG400 and Solutol HS-15, vortex to obtain a clear solution, then add the prescribed amount of pH 7.4 phosphate buffer and mix well.

[0664]

[0665]

[0666] The experimental results in the above table show that compound A107 in WO2017 / 114510A1 significantly increases impurities after being placed at 37°C for 24 hours in solutions with pH 1.2, pH 6.8, and pH 7.4, and has poor chemical stability under acidic, neutral, and weakly alkaline conditions.

[0667] Effect Example II: In vitro enzyme activity assay

[0668] In this example, the half-maximal inhibitory activity (IC 50 value)

[0669] (1) Materials and instruments:

[0670] Enzyme: Extracellular signal-regulated kinase ERK2 kinase (PV3595, Invitrogen)

[0671] Kit: Z'- Protein Kinase Assay Kit - Ser / Thr 3Peptide (PV3176, Invitrogen) Kit components: Substrate Z'-LYTE TM Ser / Thr 3Peptide(PV3200)

[0672] Phosphorylation substrate Z′-LYTE TM Ser / Thr 3Phospho-peptide(PV3215)

[0673] 5X Kinase Buffer: 250mM HEPES (pH 7.5), 50mM MgCl2,

[0674] 5mM EGTA, 0.05% BRIJ-35 (PV3189, Invitrogen)

[0675] ATP (PV3227, Invitrogen)

[0676] Development Reagent A (PV3295, Invitrogen)

[0677] Development Buffer (P3127, Invitrogen)

[0678] Stop Reagent (P3094, Invitrogen)

[0679] Microplate reader: Multifunctional microplate reader PerkinElmer

[0680] Microplate: 384-well black shallow well plate (6008269, PerkinElmer)

[0681] (2) Test method:

[0682] Z'-LYTE TM Ser / Thr 3Peptide, phosphorylation substrate Z′-LYTE TMSer / Thr 3Phospho-peptide, 1X kinase buffer (5X kinase buffer diluted 5-fold with ultrapure water), ATP, Development Reagent A, Development Buffer, and Stop Reagent were equilibrated to room temperature and prepared for sample addition. The screening concentration for detecting the effect of the compounds of the present invention on ERK enzyme activity was 1 μM (0.2 μM for positive drugs) with a 3-fold serial dilution, for a total of 7 concentrations, using 4% DMSO as a co-solvent. In a 384-well plate, 5 μL of enzyme system (50 mM HEPES pH 7.5, 1 mM EGTA, 10 mM MgCl2, 0.01% Brij-35, 4 μM substrate, 0.8 ng / μL enzyme), 2.5 μL of compound, and 2.5 μL of 400 nM ATP were added and incubated at room temperature in the dark for 60 minutes. After the reaction is completed, 5 μl of Development Reagent A diluted with Development Buffer is added to all reaction wells and incubated at room temperature in the dark for 60 min. 5 μl of Terminator is added to each well to terminate the reaction. Detect fluorescence signals (excitation light wavelength is 400 nm, emission light wavelengths are 460 nm and 528 nm).

[0683] The inhibition rate of each well was calculated based on the total activity wells and background signal wells. The data analysis method was as follows:

[0684] Phosphorylation percentage = 1 – {(emission ratio × F 100％ –C 100％ ) / [C 0％ –C 100％ + emission ratio×(F 100％ –F 0％ )]}×100

[0685] Percent Inhibitor = 100 x (1 - Percent Phosphorylation in Test Compound Wells / Percent Phosphorylation in 0% Inhibition Control Wells)

[0686] Where: Emission ratio = the ratio of 445nm emission light to 520nm emission light in the sample

[0687] F 100％ = Average fluorescence emission at 520 nm in the 100% phosphorylated substrate control wells

[0688] F 0％ = Average fluorescence emission at 520 nm in the wells with 0% phosphorylated substrate control

[0689] C 100％= Average fluorescence emission at 445 nm in the 100% phosphorylated substrate control wells

[0690] C 0％ = Average value of 445 nm emission fluorescence in the wells with 0% phosphorylation substrate control The experiment was repeated twice in parallel. IC was calculated by the inhibition value of the test compound on the kinase at a series of different concentrations 50 value.

[0691] (3) Experimental results

[0692] The inhibitory activity data of the compounds of the present invention on ERK2 kinase activity (IC 50 ) as shown in the following table, where:

[0693] A: represents the IC of the compound 50 Less than or equal to 10nM;

[0694] B: represents the IC of the compound 50 greater than 10 nM but less than 100 nM;

[0695] C: represents the IC of the compound 50 Greater than or equal to 100 nM but less than 1 μM.

[0696] Inhibitory activity data of the compounds of the present invention on ERK2 kinase activity

[0697]

[0698]

[0699] More specifically, in this example, the IC values ​​of compounds P5, P9, P10, P18, P42, P59, and P60 of the present invention are 50 The values ​​were 8.2 nM, 3.3 nM, 6.1 nM, 3.0 nM, 2.7 nM, 4.8 nM and 10 nM, respectively.

[0700] Effect Example III: In vitro cell activity study

[0701] In this example, the proliferation inhibitory activity (IC 50 value)

[0702] (1) Materials and instruments:

[0703] Cells: Human melanoma cell line A375 (CRL-1619 TM ,ATCC)

[0704] Detection reagent: Sulfolositamine B SRB (S9012, Sigma)

[0705] Culture plate: 96-well cell culture plate (3599, Corning)

[0706] Microplate reader: Full wavelength microplate reader (SpectraMax 190, Molecular Devices)

[0707] (2) Test method:

[0708] Logarithmically growing cells were seeded at an appropriate density (3500 cells / well) into 96-well cell culture plates, with 90 μL per well. After incubation overnight at 37°C in a CO2 incubator, 10 μL of various compound concentrations were added for 72 hours. Each concentration was treated in triplicate, along with a saline vehicle control and a cell-free zero well. Following treatment, cells were decultured and fixed with 10% (w / v) trichloroacetic acid (100 μL / well) for 1 hour at 4°C. Following this, cells were rinsed five times with distilled water and oven-dried. Then, 100 μL of SRB solution (4 mg / mL in 1% glacial acetic acid) was added to each well. After incubation and staining at room temperature for 15 minutes, unbound SRB was removed by rinsing five times with 1% glacial acetic acid. After oven-drying, 150 μL of 10 mM Tris solution was added to each well, and the optical density (OD) at 560 nm was measured using a SpectraMax 190 microplate reader. The inhibition rate of drug on tumor cell growth was calculated according to the following formula:

[0709]

[0710] IC 50 The values ​​were obtained by four-parameter regression using the software provided with the microplate reader. The experiment was repeated twice.

[0711] The experimental results are shown in the following table.

[0712] Comparison of cell activity data between representative compounds in WO2017 / 114510A1 and compound P9 of the present invention

[0713]

[0714] The above experimental results show that the activity of compound P9 of the present invention is significantly higher than that of the representative compound in WO2017 / 114510A1.

[0715] Effect Example IV: Permeability Study

[0716] In this example, the permeability of compound P9 of the present invention and compound A107 in WO2017 / 114510A1 was determined using a Caco-2 cell in vitro drug absorption model.

[0717] (1) Materials and instruments:

[0718] Cells: Human colon cancer Caco2 (HTB-37, ATCC)

[0719] Petri dish: 10 cm Petri dish (430167, Corning)

[0720] Millicell-24 cell culture plates (PSHT010R5, Millipore)

[0721] Buffer: PBS (14190, Invitrogen)

[0722] HEPES (H0887, Sigma)

[0723] HBSS (H8264, Sigma)

[0724] Cell culture related: High glucose DMEM medium (L0103-500, Biowest)

[0725] Fetal bovine serum (S1810-500, Biowest)

[0726] Trypsin (255200-056, Invitrogen)

[0727] Nonessential amino acids (M7145, Sigma)

[0728] Penicillin and streptomycin (B-13234, GIBCO)

[0729] Sodium pyruvate (11360-070, Invitrogen)

[0730] L-glutamine (25030-081, Invitrogen)

[0731] Related reagents: Fluorescent Yellow (L0144, Sigma)

[0732] Propranolol (P831800, Sigma)

[0733] Colchicine (C9754, Sigma)

[0734] Atenolol (A7655, Sigma)

[0735] Instrument: Liquid chromatography (Waters Acquity UPLC I-class, Waters)

[0736] Mass spectrometry (Waters xevo TQ-S MS / MS, Waters)

[0737] Resistance meter (Millicell-ERS, Thermo)

[0738] Microplate reader (Infinite Pro, Tecan)

[0739] (2) Test method:

[0740] Establishment of Caco-2 cell monolayer model

[0741] 1) Thaw Caco-2 cells. Culture in 10 cm dishes in an incubator at 37°C, 5-6% CO2, and 95% relative humidity in high-glucose DMEM supplemented with 10% fetal bovine serum, 1% glutamine, 1% non-essential amino acids, 100 U / mL penicillin, and 100 μg / mL streptomycin.

[0742] 2) When the cell density reaches 80-90%, trypsinize the cells, centrifuge, discard the supernatant, resuspend the cells in 6 ml of complete culture medium, and count three times.

[0743] 3) Centrifuge at 1000 rpm for 5 minutes to collect cells, dilute the cell suspension, and centrifuge the cells at 2 × 10 5 The concentration of 100 μg / mL was inoculated into Millicell-24 well plates, 400 μL per well, 800 μL of culture medium was added to the basolateral side, and the cells were cultured in an incubator at 37°C and 5% CO2.

[0744] 4) Change the medium 72 hours after cell inoculation and every other day thereafter. Culture for 21 days.

[0745] Evaluation of Caco-2 cell monolayers

[0746] 1) After 21 days of culture, the integrity of the Caco-2 monolayer in each well was assessed by measuring the transmembrane electrical resistance during cell growth.

[0747] 2) Marker leakage inspection

[0748] The integrity of the Caco-2 cell monolayer was verified using the fluorescent marker Lucifer Yellow. After 21 days of growth, 200 μL of Lucifer Yellow (100 μg / ml) was added to the apical side of the cell layer, and 800 μL of HBSS was added to the basolateral side. The cells were incubated in a 37°C, 5% CO2 incubator for 1.5 hours. Samples were collected and absorbance was measured at 485-535 nm. The amount of leakage was calculated, which generally did not exceed 0.4%. A blank HBSS solution was used as a blank control.

[0749] Bilateral transport experiment

[0750] Under the same conditions, drug transport from the apical side (AP side) to the basolateral side (BL side) and from the BL side to the AP side of the Caco-2 cell layer was simultaneously measured.

[0751] 1) Prepare a stock solution of the compound in DMSO at a concentration of 10 mM.

[0752] 2) Dilute the stock solution with HBSS solution to a working concentration of 20 μM.

[0753] 3) Rinse the cells three times with HBSS and measure the TEER value using a cell potential meter.

[0754] 4) Add control compound or test compound and HBSS to both sides of the cells, 400 μL / well on the AP side and 800 μL / well on the BL side.

[0755] 5) Place the cells in an incubator at 37°C and 5% CO2 for 1.5 hours, then collect samples from the AP and BL sides.

[0756] (3) Data analysis:

[0757] Apparent permeability coefficient P of drugs permeating Caco-2 cell model app (apparent permeability coefficients) are calculated according to formula (1):

[0758] Papp=(V A / (Area×Time))×([drug] acceptor / [drug] initial donor ) (1)

[0759] Where V A is the volume of the receiving side, Area is the membrane area (cm 2), Time is the reaction time, [drug] acceptor is the drug concentration at the receiving side, [drug] initial donor is the drug concentration on the dosing side.

[0760] The experimental results are shown in the following table.

[0761]

[0762] According to the measurement results of these permeability parameters, it can be seen that compared with the A107 compound in WO2017 / 114510A1, the compound P9 of the present invention has a significantly higher apparent permeability coefficient (Papp, Apical to Basal) in the Caco-2 permeability test; it is known that Papp>2×10 -6 Compound P9, a drug with good permeability (e.g., see Journal of Pharmacological and Toxicological Methods 44 (2000) 235-249), significantly exceeded this indicator, while A107 did not. A107 had a higher efflux ratio, while P9 had a lower efflux ratio. Therefore, P9 has better permeability and is expected to have better intestinal absorption and oral absorption in vivo.

[0763] Effect Example V: Solubility Determination

[0764] In this example, the thermodynamic solubility of the compounds of the present invention was determined.

[0765] (1) Reagents and materials:

[0766]

[0767]

[0768] (2) Instruments and equipment:

[0769] name source Constant temperature mixer Eppendorf centrifuge Eppendorf 5424R centrifuge, Eppendorf centrifuge Eppendorf 5810R centrifuge, Eppendorf Plate sealing machine Plate Loc plate sealer, Agilent Oscillator IKA MS3 digital oscillator, IKA Liquid chromatography Waters ACQUITY I-Class System, Waters pH meter Sartorius PB-10,Sartorius Eddy current machine MS3 digital eddy current machine, IKA analytical balance Sartorius MSE125P-100-DA Analytical Balance MS / MS systems Waters ACQUITY XEVO TQ-S (ESI source), Waters

[0770] (3) Experimental methods

[0771] Blank matrices at different pH values:

[0772] pH 7.4: DPBS

[0773] PH6.8: 9900μL HBSS+100μL HEPS+5μL 2N NaOH

[0774] pH 7.4: DPBS adjusted to pH 2.0 with 2N HCl

[0775] Experimental incubation:

[0776] Weigh approximately 3 mg of compound and add 500 μL of blank matrix respectively, and shake at 37°C for 24 h.

[0777] Sample processing:

[0778] After the incubation, the sample was centrifuged for 30 min, the supernatant was transferred to a new EP tube, and centrifuged for another 30 min.

[0779] After centrifugation, the sample was diluted 100-fold with ACN / H2O (V / V, 1:1).

[0780] ACN / H2O (V / V, 1:1) was used to prepare the linearity, and the linear concentration range was 12.5nM-1mM

[0781] Biological analysis:

[0782] All samples were mixed with water at a volume ratio of 1:1, centrifuged at 4000 rpm for 5 min, and analyzed by LC-MS / MS.

[0783] Analytical methods:

[0784] Chromatographic method:

[0785] Analytical column: Acquity BEH C18 (1.7 μm; 2.1 x 50 mm, Waters)

[0786] Mobile phase A: 0.1% FA in H2O

[0787] Mobile phase B: 0.1% FA in CAN / MEOH (9:1, V / V)

[0788] Gradient: as shown in the table below

[0789]

[0790]

[0791] MS / MS systems

[0792] Using multiple reaction monitoring (MRM) mode:

[0793] Compound Ionization mode MRM Compound P9 of the present invention ESI, positive 463.047>307.588

[0794] (4) Experimental results

[0795]

[0796] The above results show that at the three pH values ​​examined, compound P9 of the present invention has much greater solubility than the compound in WO2017 / 114510A1, which is conducive to formulation into a drug.

[0797] Effect Example VI: Pharmacokinetic Studies in Animal Models

[0798] In this example, the pharmacokinetic parameters of compound P9 of the present invention and representative compounds in WO2017 / 114510A1 in mice were determined.

[0799] (1) In vivo pharmacokinetic study plan for single intravenous (IV) and oral (PO) administration in ICR mice

[0800] (1.1) Preparation of test samples

[0801] The test sample was prepared based on the concentration of pure free base.

[0802] intravenous (IV)

[0803] Accurately weigh an appropriate amount of the test substance, add an appropriate amount of prescription excipients (5% DMSO + 5% Solutol + 90% normal saline), and after complete dissolution, prepare a dosing solution with a concentration of 0.2 mg / mL for intravenous administration.

[0804] Oral administration (PO)

[0805] An appropriate amount of the test substance was accurately weighed, and an appropriate amount of the prescription excipient (0.4% methylcellulose (viscosity: 400 cps)) was added. After thorough mixing, a dosing solution with a concentration of 1 mg / mL was prepared for oral administration.

[0806] (1.2) Animal reception and adaptation

[0807] Forty male SPF-grade ICR mice were purchased from Shanghai Xipu-Bikai Laboratory Animal Co., Ltd., of which 30 healthy ICR mice that passed the physical examination and had no abnormalities were used in this study, with animal weights ranging from 20.12 to 25.56 g.

[0808] (1.3) Animal administration

[0809] Thirty male ICR mice were used for the experiment according to the table below.

[0810]

[0811] Note*: All animals were fasted for 10-14 hours before administration and resumed feeding 2 hours after administration.

[0812] (1.4) Sample collection and processing

[0813] Blood was collected by intraorbital or carbon dioxide (CO2) euthanasia via cardiac puncture. Approximately 0.20 milliliters (mL) of blood was collected for each sample, anticoagulated with sodium heparin, and placed on ice after collection.

[0814] The collection time points for the intravenous and oral administration groups were: before administration and 5, 15 and 30 minutes, 1, 2, 4, 6, 8 and 24 hours after administration, as shown in the following table:

[0815]

[0816] After blood samples were collected, they were placed on ice and centrifuged to separate plasma (centrifugation conditions: 8000 rpm, 6 minutes, 2-8°C). The collected plasma was stored at -80°C before analysis.

[0817] Biological samples of the test article were analyzed by LC-MS / MS. The analytical method used was described in Item (1.7). The LLOQ of the sample test was 1 ng / mL. The standard curve and quality control sample analysis were performed during the sample testing process.

[0818] (1.5) Animal treatment

[0819] After the experiment, all animals were euthanized according to the institutional SOP.

[0820] (1.6) Pharmacokinetic analysis

[0821] According to the blood concentration data of the drug, WinNonlin 7.0 was used to calculate the pharmacokinetic parameters and provide AUC 0-t , AUC 0-∞ , MRT 0-∞ 、C max 、T max , and T 1 / 2 Parameters and their means and standard deviations.

[0822] For samples with concentrations below the limit of quantification, when calculating pharmacokinetic parameters, max The samples taken previously should be calculated as zero. max Subsequent sampling point samples should be calculated as no quantifiable amount (BLQ).

[0823] (1.7) Analytical Methods a. Instruments and Equipment

[0824] LC-MS / MS

[0825] Ultra-high performance liquid chromatography system (Waters, ACQUITY UPLC), including a binary solvent manager (ACQUITY UPLC Binary Solvent Manager), a sample manager (ACQUITY UPLC Autosampler Mod.), a high-throughput sample organizer (ACQUTIY UPLC Sample Organizer), and a high-temperature column oven (ACQUITY UPLC Column Heater HT).

[0826] Mass spectrometer (API 4000, Applied Biosystems, USA) with electrospray ionization (ESI) source and tandem quadrupole mass analyzer.

[0827] The data processing system was Analyst software (Applied Biosystems, USA, software version 1.5.1).

[0828] Microanalytical balance (XP26, Mettler-Toledo Instruments (Shanghai) Co., Ltd.); vortex shaker (SI-A256, Scientific Industries, Inc.); small desktop high-speed refrigerated centrifuge (5417R, Eppendorf); ultrapure water machine (Millipore); pipette (Eppendorf).

[0829] Reagents

[0830] Methanol (Burdick & Jackson, HPLC), acetonitrile (Burdick & Jackson, HPLC), formic acid (J&K), ultrapure water.

[0831] b. LC-MS / MS conditions

[0832] The liquid phase conditions are as follows:

[0833] Column: ACQUITY UPLC HSS T3 1.8 μm (50 mm × 2.10 mm)

[0834] Mobile phase:

[0835] Time (min) A (％) B(％) 0.00 80 20 0.40 10 90 0.80 10 90 0.81 80 20 1.20 80 20

[0836] A: 0.1% formic acid in water B: 0.1% formic acid in methanol

[0837] Column temperature: 40°C Autosampler temperature: 4°C

[0838] Flow rate: 500 μL / min Injection volume: 1 μL

[0839] Mass spectrometry conditions are as follows:

[0840] Scan mode: Positive ion multiple reaction monitoring mode

[0841] Ion source: electrospray ionization source Nebulization mode: electrospray

[0842] Q1 resolution: Unit Q3 resolution: Unit

[0843] Atomizing gas (Gas 1): 65psi Auxiliary heating gas (Gas 2): 65psi

[0844] Curtain gas (CUR): 35psi Collision gas (CAD): 10

[0845] Ion source voltage (IS): 5500v Ion source temperature (TEM): 550℃

[0846] c. Preparation of standard curve samples and quality control samples

[0847] Weigh a certain amount of test substance and dissolve it completely in methanol to prepare a stock solution with a concentration of 569,000 ng / mL. Take a certain amount of the stock solution and dilute it with methanol to a working solution with a concentration of 200,000 ng / mL. Take a certain amount of the 200,000 ng / mL standard solution and add it to a certain amount of blank plasma at a ratio of 1:39 to prepare a standard curve sample with a concentration of 5000 ng / mL. Take the 5000 ng / mL standard curve sample and dilute it with blank plasma in sequence to obtain standard curve samples at a concentration of 1000, 500, 100, 50, 10, 5, and 1 ng / mL, as well as quality control samples at 800, 200, and 2.5 ng / mL. The specific preparation process is shown in Table I.

[0848] Table 1 Preparation table of standard curve samples and QC samples

[0849]

[0850] Internal standard working solution: Pipette a certain amount of tolbutamide stock solution with a concentration of 767,000 ng / mL into a volumetric flask of a certain volume, dilute to the mark with methanol, and mix to prepare an internal standard working solution with a concentration of 200 ng / mL.

[0851] d. Plasma sample processing

[0852] Transfer 50 μL of sample (standard / quality control / biological sample) to a 1.5 mL centrifuge tube, add 250 μL of internal standard solution (for a blank sample without internal standard, add an equal volume of methanol), and vortex to mix. Centrifuge at 14,000 rpm for 5 minutes. Transfer 200 μL of the supernatant to the corresponding 96-well plate for LC-MS / MS analysis.

[0853] (2) Experimental results

[0854] The pharmacokinetic data of the representative compounds in WO2017 / 114510A1 and compound P9 of the present invention in mice are compared as follows.

[0855]

[0856]

[0857] According to the results of the measurement of pharmacokinetic parameters in these mice, it can be seen that compared with the compound in WO2017 / 114510A1, the compound P9 of the present invention has a significantly higher area under the drug-dose curve (AUC) and a lower in vivo clearance rate (CLz) in mice, and has good bioavailability. Therefore, it is predicted that this compound has better oral absorption and better drugability.

[0858] Formulation Example 1

[0859] Accurately weigh an appropriate amount of compound P9, add 5% DMSO + 5% Solutol + 90% saline, etc., and dissolve it completely to prepare a dosing solution with a concentration of 0.2 mg / mL. Sterile filter it for intravenous administration;

[0860] Formulation Example II

[0861] An appropriate amount of compound P9 was accurately weighed, and 0.4% methylcellulose (viscosity: 400 cps) was added to the final volume. After thorough mixing, a dosing solution with a concentration of 1 mg / mL was prepared for oral administration.

[0862] All references mentioned in this application are incorporated herein by reference in their entirety, just as if each reference were listed separately. It should be understood that after reading the disclosure of this application, those skilled in the art may make various changes or modifications to the present invention, and these equivalent forms also fall within the scope of the claims appended hereto.

Claims

1. A compound of formula (Ie) or a pharmaceutically acceptable salt thereof, Where, X1 is CH or CD; R1 is selected from H and D; R2 is selected from R3 is fluorine; R4 is -CO(CR 10 R 11 ) m R 12 ; where m is 1, R 10 and R 11 are each independently H; and R 12 yes and R5, R6, R7 and R8 are each independently selected from: -H, -D.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

3. A pharmaceutical composition comprising the compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, and optionally comprising a pharmaceutically acceptable carrier.

4. Use of the compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing and / or treating diseases associated with ERK kinase or a product as an ERK kinase inhibitor.

5. A method for preparing a compound of formula (Ie) or a pharmaceutically acceptable salt thereof according to claim 1, wherein the compound of formula (Ie) is a compound of formula C3: Among them, X1, R2, R3, R 10 、R 11 、R 12 and m as defined in claim 1, The method comprises the following steps: (a) making a compound of formula C1 With compound Carry out amide coupling reaction to generate a compound of formula C2, and (b) when the compound of formula C2 is Boc protected, deprotecting it to generate a compound of formula C3, 6. The method of claim 5, wherein the amide coupling reaction is carried out in the presence of a condensation reagent and a base in an inert solvent.

7. The method of claim 5, wherein the deprotection is carried out in the presence of an acid in an inert solvent.

8. The method of claim 6 or 7, wherein the inert solvent is selected from the group consisting of ethyl acetate, tetrahydrofuran, methyltetrahydrofuran, acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, dichloromethane, 1,2-dichloroethane, N-methyl-2-pyrrolidone, or a combination thereof.

9. The method of claim 6, wherein the condensation reagent is selected from the group consisting of: 1-hydroxybenzotriazole (HOBT), 1-hydroxy-7-azobenzotriazole (HOAT), benzotriazol-1-yl-oxytripyrrolidinium hexafluorophosphate (PyBOP), benzotriazole-1-tris(trimethylamino)-hexafluorophosphate (BOP), 1,1-carbonyldiimidazole (CDI), 1-propylphosphonic anhydride (T3P), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl), N,N-dicyclohexylcarbodiimide (DCC), acetic anhydride, acetyl chloride, oxalyl chloride, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), and O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate quaternary ammonium salt (HBTU).

10. The method of claim 6, wherein the base is selected from one or more of triethylamine, DIPEA, pyridine, 2,4-lutidine, NaOH, KOH, LiOH, Na2CO3, K2CO3, NaHCO3, Cs2CO3, Na3PO4 or K3PO4.

11. The method of claim 5, wherein the amide coupling reaction is carried out at a temperature ranging from room temperature to reflux for 0.5 to 24 hours.

12. The method of claim 7, wherein the acid is selected from the group consisting of: hydrochloric acid, sulfuric acid, trifluoroacetic acid, acetic acid, formic acid, phosphoric acid, or one or more thereof.

13. The method of claim 5, wherein the deprotection is carried out at a temperature of -10°C to 80°C for 0.5 to 24 hours.

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