Fused tricyclic cyclin-dependent kinase inhibitor as well as preparation method and medical application thereof

Abstract

The invention relates to a fused tricyclic cyclin-dependent kinase inhibitor as well as a preparation method and medical application thereof. Particularly, the structure of the fused tricyclic cyclin-dependent kinase inhibitor is shown as a formula I, and the definition of each substituent group is shown in the specification. The fused tricyclic cyclin-dependent kinase inhibitor is used for preventing and / or treating diseases related to cyclin-dependent kinase, especially cancers.

Description

[0001] This application is a divisional application of the Chinese patent application with application number 202280008155.2, application date January 28, 2022, and invention name “Fused tricyclic cyclin-dependent kinase inhibitors, preparation methods and medical uses thereof”. Technical Field

[0002] The present disclosure belongs to the field of medicine and relates to a fused tricyclic cyclin-dependent kinase inhibitor and a preparation method, a composition and medical use thereof. Background Art

[0003] Cyclin-dependent kinases (CDKs) are important cellular enzymes that play an important role in regulating eukaryotic cell division and proliferation. The cyclin-dependent kinase catalytic unit is called the regulatory subunit activation of cyclins. At least 16 mammalian cell cycle proteins have been identified (Annu. Rev. Pharmacol. Toxicol. (1999) 39: 295-312). Cyclin B / CDK1, cyclin A / CDK2, cyclin E / CDK2, cyclin D / CDK4, cyclin D / CDK6 and possible other heterodynes are important regulatory factors for cell cycle progression. Other functions of cyclin / CDK heterodynes include transcriptional regulation, DNA repair, differentiation and apoptosis (Annu. Rev. Cell. Dev. Biol. (1997) 13: 261-291).

[0004] In recent years, the greatest progress in the field of breast cancer treatment is undoubtedly the use of CDK4 / 6 alone or in combination with endocrine therapy in hormone receptor-positive advanced breast cancer. For example, palbociclib, ribociclib, and abemaciclib have been approved in combination with aromatase inhibitors for the treatment of hormone receptor (HR)-positive, human epidermal growth factor receptor 2 (HER2)-negative advanced or metastatic breast cancer in postmenopausal women, and palbociclib and abemaciclib have been approved in combination with fulvestrant for the treatment of hormone receptor (HR)-positive, human epidermal growth factor receptor 2 (HER2)-negative advanced or metastatic breast cancer in postmenopausal women after disease progression following endocrine therapy (Nature Reviews (2016) 13:417-430, J Clin Oncol 2017, 35, 2875-2884). Although CDK4 / 6 inhibitors have shown remarkable clinical efficacy in estrogen receptor (ER)-positive metastatic breast cancer, their effects, like other kinases, may be limited over time by the development of primary or acquired resistance.

[0005] Treatment with CDK4 / 6 inhibitors has been shown in the clinic to result in adverse effects, such as gastrointestinal and / or hematologic toxicities, and may lead to acquired resistance over time. Emerging data suggest that cyclin D3-CDK6 may be involved in the observed hematologic toxicities. (Malumbres et al., Mammalian Cells Cycle without the D-Type Cyclin-Dependent Kinases Cdk4 and Cdk6, (2004) Cell 118(4): 493-504; Sicinska et al., Essential Role for Cyclin D3 in Granulocyte Colony-Stimulating Factor-Driven Expansion of Neutrophil Granulocytes (2006), Mol. Cell Biol 26(21): 8052-8060; Cooper et al., A unique function for cyclin D3 in early B cell development, (2006), Nat. Immunol. 5(7): 489-497). CDK4 has been identified as a single oncogenic driver in many breast cancers. Therefore, due to the potential for higher and / or continuous dosing compared to dual CDK4 / 6 inhibitors, CDK4 selective inhibitors may provide improved safety or enhanced overall efficacy. Therefore, the development of molecules with high CDK4 selectivity has practical clinical application value. WO2019207463A discloses a class of cyclin-dependent kinase inhibitors. Summary of the Invention

[0006] The present disclosure provides a compound represented by formula I or a pharmaceutically acceptable salt thereof,

[0007]

[0008] Among them, R 1 Selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl and C 3-8 Cycloalkyl, the C 1-6 Alkyl, C 1-6 Haloalkyl and C 3-8 The cycloalkyl groups are each independently optionally substituted with one or more R a replace;

[0009] R 2 The structure of formula II: R 9 Selected from H, OH and NH2, wherein NH2 is optionally replaced by 1 or 2 R a’ or R a” replace;

[0010] R 10 Each independently selected from OH, halogen, CN, NH2, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 3-8 The cycloalkyl group and the 3-12 membered heterocycloalkyl group are each independently optionally substituted with one or more R b replace;

[0011] Q is NR 11 or O;

[0012] Or Q is CR 12 R 13 , where R 12 、R 13 The carbon atom connected to it forms a 3-12-membered NR 11 wherein N or O is a heterocycloalkyl group as a ring atom, the heterocycloalkyl group is optionally substituted by one or more R 10 replace;

[0013] R 11 Selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, SO2R c 、SO2NR d R e 、COR f 、COOR f and CONR g R h , the C 1-6 Alkyl and C 1-6 Each haloalkyl group is independently optionally selected from R a 、R b 、SO2R c 、SO2NR d R e 、COR f 、COOR f and CONR g R h substituted with one or more substituents;

[0014] m is 0, 1, or 2;

[0015] n is 0, 1, 2, 3 or 4;

[0016] p is 1, 2, or 3;

[0017] X is N or CH;

[0018] Y is N or CR 7 , R 7 Selected from H, F, Cl, CN, C 1-6 Alkyl and C 1-6 Alkoxy, the C 1-6 Alkyl and C 1-6 Each alkoxy group is independently optionally substituted with one or more R a replace;

[0019] R 3 selected from H, F, Cl, CN, CH2F, CHF2 and CF3;

[0020] R 4 Selected from H, C 1-6 Alkyl, C 1-6 Alkoxy, C3-8 cycloalkyl and 3-12 membered heterocycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 The cycloalkyl group and the 3-12 membered heterocycloalkyl group are each independently optionally substituted with one or more R b or deuterium substitution;

[0021] Z is O or CHR 8 , R 8 selected from the group consisting of hydrogen atoms, deuterium atoms and halogens;

[0022] L is -(CH2) q -, wherein -(CH2)- is optionally selected from deuterium, CN, halogen, C 1-6 Alkyl, C 3-8 Cycloalkyl, C 1-6 Alkoxy and 3-12 membered heterocycloalkyl are substituted with one or more substituents, the C 1-6 Alkyl, C 3-8 Cycloalkyl, C 1-6 Alkoxy and 3-12 membered heterocycloalkyl are each independently optionally substituted with one or more R b or deuterium substitution;

[0023] q is 1, 2, 3, or 4;

[0024] R 5 and R 6 Each independently selected from H, deuterium, CN, halogen, C 1-6 Alkyl, C 3-8 Cycloalkyl, C1-6 Alkoxy and 3-12 membered heterocycloalkyl, the C 1-6 Alkyl, C 3-8 Cycloalkyl, C 1-6 Alkoxy and 3-12 membered heterocycloalkyl are optionally each independently substituted with one or more R b or deuterium substitution;

[0025] R a and R b Each independently selected from H, OH, CN, halogen (fluorine, chlorine, bromine, iodine), C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-8 Cycloalkyl, 3-12 membered heterocycloalkyl and NR a’ R a” Substitute, the C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl are each independently optionally selected from NH2, NHCH3, N(CH3)2, halogen, OH, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl are substituted with one or more substituents;

[0026] R a’ and R a” Each independently selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl, the C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl are each independently optionally selected from NH2, NHCH3, N(CH3)2, halogen, OH, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl are substituted with one or more substituents;

[0027] or R a’ 、R a” Together with the nitrogen atom to which they are attached, they form a 3-12 membered heterocycloalkyl group, wherein the 3-12 membered heterocycloalkyl group is optionally selected from halogen, OH, C 1-6Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl are substituted with one or more substituents;

[0028] R c 、R d and R e Each independently selected from H, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 Cycloalkyl and C 1-6 alkyl halide;

[0029] R f Selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl, the C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl are each independently optionally selected from NH2, NHCH3, N(CH3)2, halogen, OH, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl are substituted with one or more substituents;

[0030] R g and R h Each independently selected from H, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl and C 3-8 Cycloalkyl, the C 1-6 Alkoxy, C 1-6 Haloalkyl and C 3-8 The cycloalkyl groups are each independently optionally substituted with one or more R a or R b replace.

[0031] In an optional embodiment, the present disclosure provides a compound of formula I or a pharmaceutically acceptable salt thereof, wherein R 1 H or C 1-6 alkyl.

[0032] In an optional embodiment, the present disclosure provides a compound of formula I or a pharmaceutically acceptable salt thereof, wherein R 1 For H.

[0033] In an optional embodiment, the present disclosure provides a compound of formula I or a pharmaceutically acceptable salt thereof, wherein R 2 Selected from

[0034]

[0035] Among them, R 9 、R 10 , m and Q are as defined in the compound represented by formula I.

[0036] In an optional embodiment, the present disclosure provides a compound of formula I or a pharmaceutically acceptable salt thereof, wherein R 2 for Among them, R 9 、R 10 , m and Q are as defined in the compound represented by formula I.

[0037] In an optional embodiment, the present disclosure provides a compound of formula I or a pharmaceutically acceptable salt thereof, wherein R 2 for where R 9 、R 10 、R 11 and m are as defined in the compound of formula I.

[0038] In an optional embodiment, the present disclosure provides a compound of formula I or a pharmaceutically acceptable salt thereof, wherein R 9 is OH or NH2, wherein the NH2 is optionally replaced by 1 or 2 R a’ or R a” Replacement, R a’ and R a” Each independently is C 1-6 alkyl.

[0039] In an optional embodiment, the present disclosure provides a compound of formula I or a pharmaceutically acceptable salt thereof, wherein R 9 For OH.

[0040] In an optional embodiment, the present disclosure provides a compound of formula I or a pharmaceutically acceptable salt thereof, wherein R 9 is NH2.

[0041] In an optional embodiment, the present disclosure provides a compound of Formula I or a pharmaceutically acceptable salt thereof, wherein X is N.

[0042] In an optional embodiment, the present disclosure provides a compound of formula I or a pharmaceutically acceptable salt thereof, wherein Y is CR 7 ; R 7 Selected from H, F, Cl and C 1-6 alkyl.

[0043] In an optional embodiment, the present disclosure provides a compound of formula I or a pharmaceutically acceptable salt thereof, wherein R 3 Selected from H, F and Cl.

[0044] In an optional embodiment, the present disclosure provides a compound of formula I or a pharmaceutically acceptable salt thereof, wherein R 4 Selected from H, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl; the C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 The cycloalkyl group and the 3-12 membered heterocycloalkyl group are each independently optionally substituted with one or more R b or deuterium substitution, R b Selected from H, OH, CN, halogen (fluorine, chlorine, bromine, iodine), C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Haloalkoxy and C 3-8 Cycloalkyl.

[0045] In an optional embodiment, the present disclosure provides a compound of formula I or a pharmaceutically acceptable salt thereof, wherein R 4 C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with one or more R b or deuterium substitution, R b Selected from OH, CN, halogen (fluorine, chlorine, bromine, iodine), C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Haloalkoxy and C 3-8 Cycloalkyl.

[0046] In an optional embodiment, the present disclosure provides a compound of formula I or a pharmaceutically acceptable salt thereof, wherein R 4 C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with one or more R b or deuterium substitution, R b Selected from OH, CN and halogen (fluorine, chlorine, bromine, iodine).

[0047] In an optional embodiment, the present disclosure provides a compound of formula I or a pharmaceutically acceptable salt thereof, wherein L is -(CH2) q -, q is selected from 1 or 2; said -(CH2)- is optionally selected from deuterium, CN, halogen, C1-6 Alkyl, C 3-8 Cycloalkyl, C 1-6 The alkoxy group and the 3- to 12-membered heterocycloalkyl group are substituted with one or more substituents.

[0048] In an optional embodiment, the present disclosure provides a compound of formula I or a pharmaceutically acceptable salt thereof, wherein L is -(CH2) q -, q is selected from 1; said -(CH2)- is optionally selected from deuterium, CN, halogen, C 1-6 Alkyl, C 3-8 Cycloalkyl, C 1-6 The alkoxy group and the 3- to 12-membered heterocycloalkyl group are substituted with one or more substituents.

[0049] In an optional embodiment, the present disclosure provides a compound of formula I or a pharmaceutically acceptable salt thereof, wherein L is -(CH2) qa -, q is selected from 1; the -(CH2)- is optionally substituted with one or more deuterium substituents.

[0050] In an optional embodiment, the present disclosure provides a compound of formula I or a pharmaceutically acceptable salt thereof, wherein R 5 and R 6 Each independently selected from H, deuterium, CN, halogen, C 1-6 Alkyl and C 3-8 Cycloalkyl.

[0051] In an optional embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula I-2 or a pharmaceutically acceptable salt thereof,

[0052]

[0053] where R 9 is OH or NH2, wherein the NH2 is optionally replaced by 1 or 2 R a’ or R a” Replacement, R a’ and R a” Each independently is C 1-6 alkyl;

[0054] R 10 Each independently selected from H, OH, halogen, CN, NH2, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl and C 1-6 haloalkoxy;

[0055] m is 0, 1, or 2;

[0056] R 11 Selected from SO2Rc 、SO2NR d R e 、COR f 、COOR f and CONR g R h ;

[0057] R c 、R d and R e Each independently selected from H, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 Cycloalkyl and C 1-6 alkyl halide;

[0058] R f Selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl;

[0059] R g 、R h Each independently selected from H, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl and C 3-8 Cycloalkyl;

[0060] R 7 Selected from H, F, Cl and C 1-6 alkyl;

[0061] R 3 selected from H, F and Cl;

[0062] R 4 Selected from H, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl; the C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 The cycloalkyl group and the 3-12 membered heterocycloalkyl group are each independently optionally substituted with one or more R b or deuterium substitution, R b Selected from H, OH, CN and halogen (fluorine, chlorine, bromine, iodine);

[0063] R 5 and R 6 Each independently selected from H, deuterium, CN, halogen, C 1-6 Alkyl and C 3-8 Cycloalkyl;

[0064] Z is O or CHR8 , R 8 is selected from the group consisting of a hydrogen atom, a deuterium atom and a halogen.

[0065] In an optional embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula I-2 or a pharmaceutically acceptable salt thereof 2, wherein Z is O.

[0066] In an optional embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula I-2 or a pharmaceutically acceptable salt thereof, wherein Z is CHR 8 , R 8 is selected from the group consisting of a hydrogen atom, a deuterium atom and a halogen.

[0067] In an optional embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula I-2 or a pharmaceutically acceptable salt thereof,

[0068] where R 9 is OH or NH2, wherein the NH2 is optionally replaced by 1 or 2 R a’ or R a” Replacement, R a’ and R a” Each independently is C 1-6 alkyl;

[0069] m is 0;

[0070] R 11 SO2R c ;

[0071] R c Selected from H, C 1-6 Alkyl and C 1-6 alkyl halide;

[0072] R 7 Selected from H, F, Cl and C 1-6 alkyl;

[0073] R 3 selected from H, F and Cl;

[0074] R 4 Selected from H, C 1-6 Alkyl and C 1-6 Alkoxy; the C 1-6 Alkyl and C 1-6 Each alkoxy group is independently optionally substituted with one or more R b or deuterium substitution; R b Selected from H, OH, CN and halogen (fluorine, chlorine, bromine, iodine);

[0075] R 5 and R6 Each independently selected from H, deuterium, CN, halogen and C 1-6 alkyl.

[0076] In an optional embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula I-2 or a pharmaceutically acceptable salt thereof,

[0077] where R 9 for OH;

[0078] m is 0;

[0079] R 11 SO2R c ;

[0080] R c C 1-6 Alkyl or C 1-6 alkyl halide;

[0081] R 7 is F or Cl;

[0082] R 3 is F or Cl;

[0083] R 4 H or C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with one or more R b or deuterium substitution, R b Selected from H, OH, CN and halogen (fluorine, chlorine, bromine, iodine);

[0084] R 5 and R 6 Each is independently H or C 1-6 alkyl.

[0085] In an optional embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula I-2 or a pharmaceutically acceptable salt thereof,

[0086] where R 9 for OH;

[0087] m is 0;

[0088] R 11 SO2R c ;

[0089] R c is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl;

[0090] R 7 is F or Cl;

[0091] R 3 is F or Cl;

[0092] R 4 H or C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with one or more R b or deuterium substitution, R b is H or OH;

[0093] R 5 and R 6 Each is independently selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl.

[0094] In an optional embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula I-2 or a pharmaceutically acceptable salt thereof,

[0095] where R 9 for OH;

[0096] m is 0;

[0097] R 11 SO2R c ;

[0098] R c is selected from methyl, ethyl and n-propyl;

[0099] R 7 is F or Cl;

[0100] R 3 is F or Cl;

[0101] R 4 is selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl, wherein the methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl are each independently optionally replaced by one or more R b Replacement, R b is H or OH;

[0102] R 5 and R 6 Each is independently selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl.

[0103] In an optional embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula I-2 or a pharmaceutically acceptable salt thereof,

[0104] where R 9 for OH;

[0105] m is 0;

[0106] R 11 SO2R c ;

[0107] R c is methyl;

[0108] R 7 is F or Cl;

[0109] R 3 is F or Cl;

[0110] R 4 is selected from H, methyl, ethyl, n-propyl and isopropyl;

[0111] R 5 and R 6 Each is independently selected from H, methyl, ethyl, n-propyl and isopropyl.

[0112] In an optional embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula I-3 or a pharmaceutically acceptable salt thereof,

[0113]

[0114] where R 9 is OH or NH2, wherein the NH2 is optionally replaced by 1 or 2 R a’ or R a” Replacement, R a’ and R a” Each independently is C 1-6 alkyl;

[0115] R 10 Each independently selected from OH, halogen, CN, NH2, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl and C 1-6 haloalkoxy;

[0116] m is 0, 1, or 2;

[0117] R 7 Selected from H, F, Cl and C 1-6 alkyl;

[0118] R 3 selected from H, F and Cl;

[0119] R 4 Selected from H, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl, the C 1-6Alkyl, C 1-6 Alkoxy, C3-8 cycloalkyl and 3-12 membered heterocycloalkyl are each independently optionally substituted with one or more R b or deuterium substitution; R b Selected from H, OH, CN and halogen (fluorine, chlorine, bromine, iodine);

[0120] R 5 and R 6 Each independently selected from H, deuterium, CN, halogen, C 1-6 Alkyl and C 3-8 Cycloalkyl;

[0121] Z is O or CHR 8 , R 8 is selected from the group consisting of a hydrogen atom, a deuterium atom and a halogen.

[0122] In an optional embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula I-3 or a pharmaceutically acceptable salt thereof, wherein Z is O.

[0123] In an optional embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula I-3 or a pharmaceutically acceptable salt thereof, wherein Z is CHR 8 , R 8 is a hydrogen atom, a deuterium atom or a halogen.

[0124] In an optional embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula I-3 or a pharmaceutically acceptable salt thereof,

[0125] where R 9 is OH or NH2, wherein the NH2 is optionally replaced by 1 or 2 R a’ or R a” Replacement, R a’ and R a” Each independently is C 1-6 alkyl;

[0126] m is 0;

[0127] R 7 Selected from H, F, Cl and C 1-6 alkyl;

[0128] R 3 selected from H, F and Cl;

[0129] R 4 Selected from H, C 1-6 Alkyl and C 1-6 Alkoxy; the C 1-6 Alkyl and C 1-6Each alkoxy group is independently optionally substituted with one or more R b or deuterium substitution, R b Selected from H, OH, CN and halogen (fluorine, chlorine, bromine, iodine);

[0130] R 5 and R 6 Each independently selected from H, deuterium, CN, halogen and C 1-6 alkyl.

[0131] In an optional embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula I-3 or a pharmaceutically acceptable salt thereof,

[0132] where R 9 for OH;

[0133] m is 0;

[0134] R 7 is F or Cl;

[0135] R 3 is F or Cl;

[0136] R 4 H or C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with one or more R b or deuterium substitution, R b Selected from H, OH, CN and halogen (fluorine, chlorine, bromine, iodine);

[0137] R 5 and R 6 Each is independently H or C 1-6 alkyl.

[0138] In an optional embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula I-3 or a pharmaceutically acceptable salt thereof,

[0139] where R 9 for OH;

[0140] m is 0;

[0141] R 7 is F or Cl;

[0142] R 3 is F or Cl;

[0143] R 4 H or C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with one or more R b or deuterium substitution, Rb is H or OH;

[0144] R 5 and R 6 Each is independently selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl.

[0145] In an optional embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula I-3 or a pharmaceutically acceptable salt thereof,

[0146] where R 9 for OH;

[0147] m is 0;

[0148] R 7 is F or Cl;

[0149] R 3 For F Hu Cl;

[0150] R 4 is selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl, wherein the methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl are each independently optionally replaced by one or more R b Replacement; R b is H or OH;

[0151] R 5 and R 6 Each is independently selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl.

[0152] In an optional embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula I-3 or a pharmaceutically acceptable salt thereof,

[0153] where R 9 for OH;

[0154] m is 0;

[0155] R 7 is F or Cl;

[0156] R 3 is F or Cl;

[0157] R 4 is selected from H, methyl, ethyl, n-propyl and isopropyl;

[0158] R 5 and R 6 Each is independently selected from H, methyl, ethyl, n-propyl and isopropyl.

[0159] In an optional embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is selected from

[0160]

[0161]

[0162]

[0163] Another aspect of the present disclosure provides an isotope substitution of the compound represented by the aforementioned formula I, I-2, I-3 or a pharmaceutically acceptable salt thereof. In an optional embodiment, the isotope substitution is a deuterium atom substitution.

[0164] In an optional embodiment, the compound of Formula I or a pharmaceutically acceptable salt thereof, has an abundance of deuterium atoms greater than 20%.

[0165] In an optional embodiment, the compound of Formula I or a pharmaceutically acceptable salt thereof, has an abundance of deuterium atoms greater than 50%.

[0166] In an optional embodiment, the compound of Formula I or a pharmaceutically acceptable salt thereof, has an abundance of deuterium atoms greater than 90%.

[0167] In an optional embodiment, the compound of Formula I or a pharmaceutically acceptable salt thereof, has an abundance of deuterium atoms greater than 95%.

[0168] The present disclosure also provides a method for preparing a compound of formula I, comprising the steps of reacting a compound of formula IB with a compound of formula IC to form a compound of formula I.

[0169]

[0170] Among them, LG 1 is a leaving group, wherein the leaving group is preferably a halogen, a sulfonate, a boronic acid, and a borate;

[0171] X, Y, Z, L, R 1 、R 2 、R 3 、R 4 、R 5 and R 6 As defined in the compound represented by formula I.

[0172] In some embodiments, the reaction is carried out in the presence of a catalyst, which is metallic palladium or metallic nickel.

[0173] In some embodiments, the catalyst is selected from palladium / carbon, Raney nickel, tetrakistriphenylphosphine palladium, palladium dichloride, palladium acetate, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium, 1,1'-[1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium, tris(dibenzylideneacetone)dipalladium and 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl, preferably [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium and 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl.

[0174] Another aspect of the present disclosure provides a compound represented by Formula IB or a pharmaceutically acceptable salt thereof,

[0175]

[0176] Among them, LG 1 is a leaving group selected from halogen, sulfonate, boronic acid and borate; X, Y, Z, L, R 3 、R 4 、R 5 、R 6 As defined in the compound represented by formula I.

[0177] The present disclosure also provides a pharmaceutical composition comprising at least one therapeutically effective amount of the compound represented by the aforementioned formula I, I-2, or I-3, or a pharmaceutically acceptable salt or the aforementioned isotope substitute, and a pharmaceutically acceptable excipient.

[0178] In some embodiments, the unit dose of the pharmaceutical composition is 0.001 mg-1000 mg.

[0179] In certain embodiments, the pharmaceutical composition contains 0.01-99.99% of the compound represented by the aforementioned formula I, I-2, I-3, or a pharmaceutically acceptable salt thereof, or the aforementioned isotope substitute, based on the total weight of the composition.

[0180] In certain embodiments, the pharmaceutical composition contains 0.1-99.9% of the compound represented by the aforementioned formula I, I-2, I-3, or a pharmaceutically acceptable salt thereof, or the aforementioned isotopic substitution.

[0181] In certain embodiments, the pharmaceutical composition contains 0.5%-99.5% of the compound represented by the aforementioned formula I, I-2, I-3, or a pharmaceutically acceptable salt thereof, or the aforementioned isotopic substitution.

[0182] In certain embodiments, the pharmaceutical composition contains 1%-99% of the compound represented by the aforementioned formula I, I-2, I-3, or a pharmaceutically acceptable salt thereof, or the aforementioned isotopic substitution.

[0183] In certain embodiments, the pharmaceutical composition contains 2%-98% of the compound represented by the aforementioned formula I, I-2, I-3, or a pharmaceutically acceptable salt thereof, or the aforementioned isotope substitute.

[0184] In certain embodiments, the pharmaceutical composition contains 0.01%-99.99% of a pharmaceutically acceptable excipient based on the total weight of the composition.

[0185] In certain embodiments, the pharmaceutical composition contains 0.1%-99.9% of a pharmaceutically acceptable excipient.

[0186] In certain embodiments, the pharmaceutical composition contains 0.5%-99.5% of a pharmaceutically acceptable excipient.

[0187] In certain embodiments, the pharmaceutical composition contains 1%-99% of a pharmaceutically acceptable excipient.

[0188] In certain embodiments, the pharmaceutical composition contains 2%-98% of a pharmaceutically acceptable excipient.

[0189] The present disclosure also provides a method for preventing and / or treating diseases related to cyclin-dependent kinases, which comprises administering a therapeutically effective amount of the compound represented by the aforementioned formula I, I-2, or I-3, or a pharmaceutically acceptable salt thereof, or the aforementioned isotope substitute, or the aforementioned pharmaceutical composition to a patient in need thereof.

[0190] The present disclosure also provides a method for preventing and / or treating cancer, comprising administering a therapeutically effective amount of the compound represented by the aforementioned formula I, I-2, or I-3, or a pharmaceutically acceptable salt thereof, or the aforementioned isotope substitute, or the aforementioned pharmaceutical composition to a patient in need thereof.

[0191] The present disclosure also provides the use of the compounds represented by the aforementioned formulas I, I-2, and I-3, or their pharmaceutically acceptable salts or the aforementioned isotopic substitutes, or the aforementioned pharmaceutical compositions in the preparation of drugs for preventing and / or treating diseases associated with cyclin-dependent kinases.

[0192] The present disclosure also provides the use of the compounds represented by the aforementioned formulas I, I-2, and I-3, or their pharmaceutically acceptable salts or the aforementioned isotope substitutes, or the aforementioned pharmaceutical compositions in the preparation of drugs for preventing and / or treating cancer.

[0193] In an alternative embodiment, the cyclin-dependent kinase-related disease is selected from cell proliferative diseases, cancer and immune diseases.

[0194] The cancer in the present disclosure is selected from breast cancer, ovarian cancer, bladder cancer, uterine cancer, prostate cancer, lung cancer (including NSCLC, SCLC, squamous cell carcinoma or adenocarcinoma), esophageal cancer, head and neck cancer, intestinal cancer, kidney cancer (including RCC), liver cancer (including HCC), pancreatic cancer, gastric cancer and thyroid cancer.

[0195] In alternative embodiments, the cyclin-dependent kinase described in the present disclosure is CDK4.

[0196] Another aspect of the present disclosure provides a use of the compound represented by the aforementioned formula I, I-2, or I-3, or a pharmaceutically acceptable salt thereof, or the aforementioned isotope-substituted product as a medicine.

[0197] The compounds of the aforementioned formulas I, I-2, and I-3, or their pharmaceutically acceptable salts or isotope substitutes, or the aforementioned pharmaceutical compositions, provided by the present disclosure, reduce gastrointestinal and / or hematological toxicity.

[0198] On the other hand, the pharmaceutically acceptable salts of the compounds described in the present disclosure are inorganic salts or organic salts.

[0199] The compounds of the present disclosure may exist in specific geometric or stereoisomeric forms. The present disclosure contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present disclosure. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All of these isomers and their mixtures are included within the scope of the present disclosure. The compounds of the present disclosure containing asymmetric carbon atoms can be isolated in optically pure form or in racemic form. Optically pure forms can be resolved from racemic mixtures or synthesized by using chiral starting materials or chiral reagents.

[0200] Optically active (R)- and (S)-isomers, as well as D and L isomers, can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present disclosure is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are separated by conventional methods known in the art, and then the pure enantiomer is recovered. In addition, the separation of enantiomers and diastereomers is typically accomplished using chromatography, which employs a chiral stationary phase and is optionally combined with a chemical derivatization method (e.g., carbamate formation from an amine).

[0201] In the chemical structures of the compounds disclosed herein, the bond " " indicates that the configuration is not specified, i.e. if chiral isomers exist in the chemical structure, the bond " ” can be "or" ”, or both "and" "Two configurations. Key" ” indicates unspecified configuration, including cis (E) or trans (Z) configuration.

[0202] Compounds and intermediates of the present disclosure can also exist in different tautomeric forms, and all such forms are included in the scope of the present disclosure. The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that can interconvert via a low energy barrier. For example, proton tautomers (also referred to as prototransfer tautomers) include interconversions via proton migration, such as keto-enol and imine-enamine, lactam-lactim isomerization. The lactam-lactim equilibrium example is between A and B as shown below.

[0203]

[0204] All compounds in this disclosure can be drawn as either Form A or Form B. All tautomeric forms are within the scope of this disclosure. The naming of the compounds does not exclude any tautomers.

[0205] The present disclosure also includes isotopically labeled compounds of the present disclosure that are identical to those described herein, but where one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into the compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as 2 H.3 H. 11 C. 13 C. 14 C. 13 N. 15 N. 15 O. 17 O. 18 O. 31 P. 32 P. 35 S. 18 F. 123 I. 125 I and 36 Cl et al.

[0206] Unless otherwise stated, when a position is specifically designated as deuterium (D), the position is understood to have at least 1000 times the abundance of deuterium greater than the natural abundance of deuterium (which is 0.015%) (i.e., at least 10% deuterium incorporation). In the example, the compound has a natural abundance greater than deuterium that can be at least 1000 times the abundance of deuterium, at least 2000 times the abundance of deuterium, at least 3000 times the abundance of deuterium, at least 4000 times the abundance of deuterium, at least 5000 times the abundance of deuterium, at least 6000 times the abundance of deuterium or more abundant deuterium. The disclosure also includes various deuterated forms of formula (I) compounds. Each available hydrogen atom connected to a carbon atom can be independently replaced by a deuterium atom. Those skilled in the art can synthesize deuterated forms of formula (I) compounds with reference to relevant literature. Commercially available deuterated starting materials may be used in the preparation of deuterated forms of the compounds of formula (I), or they may be synthesized using conventional techniques using deuterated reagents, including but not limited to deuterated borane, trideuterated borane in tetrahydrofuran, deuterated lithium aluminum hydride, deuterated iodoethane, deuterated iodomethane, and the like.

[0207] Explanation of terms:

[0208] "Pharmaceutically acceptable excipients" include, but are not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavoring agent, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier approved by the U.S. Food and Drug Administration for use in humans or domestic animals.

[0209] "Alkyl" refers to a saturated aliphatic hydrocarbon group, including straight and branched chain groups of 1 to 20 carbon atoms. Alkyl groups containing 1 to 12 carbon atoms are preferred, and alkyl groups containing 1 to 6 carbon atoms are more preferred. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, and various branched chain isomers thereof. Alkyl groups can be substituted or unsubstituted. When substituted, the substituents can be substituted at any available point of attachment, preferably one or more of the following groups, independently selected from halogen, hydroxy, oxo, nitro, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-7 Cycloalkyl, 3-12 membered heterocyclic group, etc.

[0210] "Alkenyl" includes branched and straight chain alkenes having 2 to 12 carbon atoms or alkenes containing aliphatic hydrocarbon groups. For example, "C 2-6 "Alkenyl" means an alkenyl group having 2, 3, 4, 5 or 6 carbon atoms. Examples of alkenyl groups include, but are not limited to, vinyl, allyl, 1-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methylbut-2-enyl, 3-methylbut-1-enyl, 1-pentenyl, 3-pentenyl and 4-hexenyl. Alkenyl groups may be substituted or unsubstituted. When substituted, the substituents may be substituted at any available point of attachment, preferably one or more of the following groups, independently selected from halogen, hydroxy, oxo, nitro, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-7 Cycloalkyl, 3-12 membered heterocyclic group, etc.

[0211] "Alkynyl" includes branched and straight chain alkynyl or alkenyl containing aliphatic hydrocarbon groups having 2 to 12 carbon atoms, or if a specific number of carbon atoms is specified, that specific number is intended. Examples include ethynyl, propynyl (e.g., 1-propynyl, 2-propynyl), 3-butynyl, pentynyl, hexynyl, and 1-methylpent-2-ynyl. Alkynyl groups may be substituted or unsubstituted. When substituted, the substituents may be substituted at any available point of attachment, preferably one or more of the following groups, independently selected from halogen, hydroxy, oxo, nitro, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-7 Cycloalkyl, 3-12 membered heterocyclic group, etc.

[0212] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent, wherein the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, and more preferably 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like; polycyclic cycloalkyls include spirocyclic, fused, and bridged cycloalkyls.

[0213] The cycloalkyl ring may be fused to an aryl, heteroaryl or heterocycloalkyl ring, wherein the ring attached to the parent structure is a cycloalkyl, non-limiting examples of which include indanyl, tetrahydronaphthyl, benzocycloheptyl, etc. The cycloalkyl may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups, which are independently selected from halogen, hydroxy, oxo, nitro, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-7 Cycloalkyl, 3-12 membered heterocyclic group, etc.

[0214] The term "heterocyclyl" also referred to as heterocycloalkyl, refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent containing 3 to 20 ring atoms, one or more of which is selected from nitrogen, oxygen or S(O) m (wherein m is an integer from 0 to 2) heteroatoms, but excluding the ring portion of -OO-, -OS- or -SS-, the remaining ring atoms are carbon. Preferably, it contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably, it contains 3 to 8 ring atoms. Non-limiting examples of monocyclic heterocyclic groups include pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc. Polycyclic heterocyclic groups include spirocyclic, fused ring and bridged ring heterocyclic groups. Non-limiting examples of "heterocyclic group" include:

[0215]

[0216] The heterocyclyl ring may be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring attached to the parent structure is a heterocyclyl, non-limiting examples of which include:

[0217] wait.

[0218] The heterocyclic group may be optionally substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups independently selected from halogen, hydroxy, oxo, nitro, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-7 Cycloalkyl, 3-12 membered heterocyclic group, etc.

[0219] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., rings sharing adjacent pairs of carbon atoms) group having a conjugated π electron system, preferably 6- to 12-membered, such as phenyl and naphthyl. The aryl ring may be fused to a heteroaryl, heterocyclyl, or cycloalkyl ring, wherein the ring attached to the parent structure is the aryl ring, non-limiting examples of which include:

[0220]

[0221] Aryl may be substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups independently selected from halogen, hydroxy, oxo, nitro, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-7 Cycloalkyl, 3-12 membered heterocyclic group, etc., preferably phenyl.

[0222] The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. The heteroaryl group is preferably 6 to 12-membered, more preferably 5-membered or 6-membered. For example, non-limiting examples include: imidazolyl, furyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, pyrrolyl, tetrazolyl, pyridyl, pyrimidinyl, thiadiazole, pyrazine, etc.

[0223] The heteroaryl ring may be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring that is attached to the parent structure is a heteroaryl ring, non-limiting examples of which include:

[0224]

[0225] Heteroaryl may be optionally substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups independently selected from halogen, hydroxy, oxo, nitro, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-7 Cycloalkyl, 3-12 membered heterocyclic group, etc.

[0226] The term "alkoxy" refers to -O-(alkyl) and -O-(unsubstituted cycloalkyl), wherein alkyl is as defined above. Non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy. Alkoxy may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from halogen, hydroxy, oxo, nitro, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-7Cycloalkyl, 3-12 membered heterocyclic group, etc.

[0227] The term "hydroxy" refers to an -OH group.

[0228] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0229] The term "amino" refers to -NH2.

[0230] The term "cyano" refers to -CN.

[0231] The term "nitro" refers to -NO2.

[0232] The term "oxo" refers to a =0 substituent.

[0233] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "a heterocyclic group optionally substituted with an alkyl group" means that an alkyl group may but need not be present, and that the description includes instances where the heterocyclic group is substituted with an alkyl group and instances where the heterocyclic group is not substituted with an alkyl group.

[0234] "Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3 hydrogen atoms, in a group are replaced independently of one another by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and a person skilled in the art can determine (by experiment or theory) which substitutions are possible or impossible without undue effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated (e.g., olefinic) bond.

[0235] A "pharmaceutical composition" refers to a mixture containing one or more compounds described herein, or their physiologically and pharmaceutically acceptable salts or prodrugs, together with other chemical components, and other components such as physiologically and pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, promote absorption of the active ingredient, and thereby exert its biological activity. DETAILED DESCRIPTION

[0236] The present disclosure is further described below with reference to embodiments, but these embodiments do not limit the scope of the present disclosure.

[0237] Experimental methods in the examples disclosed herein that do not specify specific conditions are generally performed under conventional conditions or according to the conditions recommended by the raw material or product manufacturers. Reagents that do not specify their specific sources are conventional reagents purchased from the market.

[0238] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). -6The unit of ppm is given. NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD3OD), and tetramethylsilane (TMS) as the internal standard.

[0239] MS was measured using a Shimadzu 2010 Mass Spectrometer or an Agilent 6110A MSD mass spectrometer.

[0240] HPLC determination was performed using a Shimadzu LC-20A systems, Shimadzu LC-2010HT series, or Agilent 1200LC high pressure liquid chromatograph (Ultimate XB-C18 3.0*150 mm column or Xtimate C18 2.1*30 mm column).

[0241] Chiral HPLC analysis was performed using Chiralpak IC-3 100×4.6mm ID, 3um, Chiralpak AD-3 150×4.6mm ID, 3um, Chiralpak AD-3 50×4.6mm ID, 3um, Chiralpak AS-3 150×4.6mm ID, 3um, Chiralpak AS-3 100×4.6mm ID, 3μm, ChiralCel OD-3 150×4.6mmI.D., 3um, Chiralcel OD-3 100×4.6mm ID, 3μm, ChiralCel OJ-H 150×4.6mm ID, 5um, Chiralcel OJ-3 150×4.6mm ID, 3um columns;

[0242] The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate. The specification of the silica gel plate used in thin layer chromatography (TLC) is 0.15mm~0.2mm, and the specification used for thin layer chromatography separation and purification products is 0.4mm~0.5mm.

[0243] Column chromatography generally uses Yantai Huanghai silica gel 100-200 mesh, 200-300 mesh or 300-400 mesh silica gel as the carrier.

[0244] Chiral preparative columns used were DAICEL CHIRALPAK IC (250 mm*30 mm, 10 μm) or Phenomenex-Amylose-1 (250 mm*30 mm, 5 μm).

[0245] The CombiFlash rapid preparation instrument used was Combiflash Rf150 (TELEDYNE ISCO).

[0246] Average kinase inhibition rate and IC 50 The values ​​were determined using a NovoStar microplate reader (BMG, Germany).

[0247] The known starting materials disclosed herein can be synthesized by methods known in the art, or can be purchased from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, AccelaChemBio Inc, Darui Chemicals, and other companies.

[0248] Unless otherwise specified in the examples, all reactions can be carried out under an argon atmosphere or a nitrogen atmosphere.

[0249] Argon atmosphere or nitrogen atmosphere means that the reaction bottle is connected to an argon or nitrogen balloon with a capacity of about 1 L.

[0250] Hydrogen atmosphere means that the reaction bottle is connected to a hydrogen balloon with a capacity of about 1L.

[0251] The pressurized hydrogenation reaction uses a Parr 3916EKX hydrogenator and a Qinglan QL-500 hydrogen generator or an HC2-SS hydrogenator.

[0252] The hydrogenation reaction is usually carried out by evacuating the chamber and filling it with hydrogen, and the operation is repeated three times.

[0253] A CEM Discover-S 908860 microwave reactor was used for the microwave reaction.

[0254] Unless otherwise specified in the examples, the solution refers to an aqueous solution.

[0255] Unless otherwise specified in the examples, the reaction temperature is room temperature, 20°C to 30°C.

[0256] The reaction progress in the examples was monitored using thin layer chromatography (TLC).

[0257] The developing solvent used in the reaction, the eluent system for column chromatography and the developing solvent system for thin layer chromatography used to purify the compound, and the volume ratio of the solvents are adjusted according to the polarity of the compound, and can also be adjusted by adding a small amount of alkaline or acidic reagents such as triethylamine and acetic acid.

[0258] Example 1

[0259] (3S,4R)-4-((5-fluoro-4-(8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylene-6-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol Isomer 1

[0260] (3S,4R)-4-((5-fluoro-4-(-8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylene-6-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol Isomer 2

[0261]

[0262] first step

[0263] 6-Bromo-4-fluoro-2,3-dinitrophenol 1b

[0264] Compound 1a (3.5 g, 14.8 mmol) was dissolved in 16 mL of dichloromethane. A solution of nitric acid in dichloromethane (2 mol / L, 16 mL) was added. The reaction was allowed to react at room temperature for 20 minutes. The reaction solution was poured into 50 mL of ice water, the organic phase was separated, and the aqueous phase was extracted with dichloromethane (50 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was collected. The filtrate was concentrated under reduced pressure to obtain the title compound 1b (3.8 g, yield: 91%).

[0265] MS (ESI) m / z 279.0, 281.0 [MH] -

[0266] Step 2

[0267] 2-Amino-6-bromo-4-fluoro-3-nitrophenol 1c

[0268] Compound 1b (3.8 g, 13.5 mmol) was dissolved in 60 mL of methanol. 25 mL of concentrated hydrochloric acid was added, and stannous chloride dihydrate (9.2 g, 40.6 mmol) was added portionwise. The reaction mixture was allowed to react at room temperature for 20 minutes. The reaction solution was concentrated, 100 mL of saturated sodium bicarbonate solution was added, and the mixture was extracted with ethyl acetate (100 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was collected. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography using petroleum ether and ethyl acetate as eluents to obtain the title compound 1c (2.0 g, 59% yield).

[0269] MS (ESI) m / z 249.1, 251.1 [MH] -

[0270] Step 3

[0271] 8-Bromo-6-fluoro-3-methyl-5-nitro-3,4-dihydro-2H-benzo[b][1,4]oxazine 1d

[0272] Compound 1c (200 mg, 0.8 mmol) was dissolved in 4 mL of acetone and cooled to 0°C. Potassium carbonate (121 mg, 0.9 mmol) and bromoacetone (120 mg, 0.9 mmol) were added at 0°C and allowed to react at room temperature for 2 hours. 20 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was collected. The filtrate was concentrated under reduced pressure, and the residue was dissolved in 5 mL of tetrahydrofuran. 0.5 mL of trifluoroacetic acid was added, and sodium cyanoborohydride (75 mg, 1.2 mmol) was added portionwise. The mixture was allowed to react at room temperature for 2 hours. The reaction mixture was poured into 20 mL of saturated sodium bicarbonate solution and extracted with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was collected. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography using petroleum ether and ethyl acetate as eluents to obtain the title compound 1d (160 mg, 69% yield).

[0273] MS (ESI) m / z 291.2, 293.2 [M+H] +

[0274] Step 4

[0275] 8-Bromo-6-fluoro-3-methyl-3,4-dihydro-2H-benzo[b][1,4]oxazin-5-amine 1e

[0276] Compound 1d (160 mg, 0.5 mmol) was dissolved in 4 mL of methanol. 2 mL of concentrated hydrochloric acid was added, and stannous chloride dihydrate (496 mg, 2.2 mmol) was added portionwise. The reaction was allowed to react at room temperature for 2 hours. The reaction solution was poured into 20 mL of saturated sodium bicarbonate solution and extracted with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was collected. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography using petroleum ether and ethyl acetate as eluents to obtain the title compound 1e (105 mg, yield: 73%).

[0277] MS (ESI) m / z 261.3, 263.3 [M+H] +

[0278] Step 5

[0279] 6-Bromo-8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylene 1f

[0280] Compound 1e (105 mg, 0.4 mmol) was dissolved in 2 mL of concentrated hydrochloric acid. 0.5 mL of acetic acid was added and the mixture was allowed to react at 120°C for 2 hours. The reaction solution was concentrated, 30 mL of saturated sodium bicarbonate solution was added, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was collected. The filtrate was concentrated under reduced pressure, and the residue was purified by C-18 reverse phase chromatography to obtain the title compound 1f (17 mg, yield: 15%).

[0281] MS (ESI) m / z 241.3, 243.3 [M+H] +

[0282] Step 6

[0283] 6-(2-chloro-5-fluoropyrimidin-4-yl)-8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylene 1g

[0284] Under a nitrogen atmosphere, compound 1f (40 mg, 0.14 mmol), pinacol diboron (57 mg, 0.22 mmol), potassium acetate (29 mg, 0.29 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (21 mg, 0.03 mmol) were dissolved in 2 mL of 1,4-dioxane. The mixture was reacted at 100°C for 1 hour. After cooling to room temperature, 1,3,5,7-tetramethyl-6-phenyl-2,4,8-trioxa-6-phosphoryladamantane (8 mg, 0.03 mmol), potassium carbonate (41 mg, 0.29 mmol), 2,4-dichloro-5-fluoropyrimidine (35 mg, 0.21 mmol), tris(dibenzylideneacetone)dipalladium (26 mg, 0.03 mmol), and 0.5 mL of water were added and the reaction was continued at 80°C for 1 hour. The reaction solution was cooled to room temperature, filtered, and the filtrate was collected and concentrated under reduced pressure. The residue was purified by C-18 reverse phase chromatography to obtain 1 g (18 mg, yield: 38%) of the title compound.

[0285] MS (ESI) m / z 337.2 [M+H] +

[0286] Step 7

[0287] (3S,4R)-4-((5-fluoro-4-(8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylene-6-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol Isomer 1

[0288] (3S,4R)-4-((5-fluoro-4-(-8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylene-6-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol Isomer 2

[0289] Under a nitrogen atmosphere, compound 1g (18 mg, 0.05 mmol), (3S,4R)-4-aminotetrahydro-2H-pyran-3-ol (7.5 mg, 0.06 mmol), (S)-(-)-2,2"-bis(diphenylphosphino)-1,1"-binaphthyl (6.2 mg, 0.01 mmol), and palladium acetate (2.2 mg, 0.01 mmol) were dissolved in 2 mL of tetrahydrofuran. Cesium carbonate (41 mg, 0.13 mmol) was added, and the reaction was allowed to proceed at 80°C for 1 hour.

[0290] The reaction solution was cooled to room temperature, filtered, and the filtrate was collected. The residue was concentrated under reduced pressure and purified by C-18 reverse phase chromatography to give a crude mixture. The crude product was separated by chiral separation (column: DAICEL CHIRALPAK AD (250 mm*30 mm, 10 μm), conditions: 45% EtOH (0.1% NH3·H2O) in CO2; flow rate: 80 mL / min) to give isomer 1 (3.8 mg, yield: 17%) and isomer 2 (4.4 mg, yield: 20%).

[0291] Analytical methods

[0292] Column: DAICEL CHIRALPAK AD-3 (150 mm*4.6 mm, 3 μm);

[0293] Conditions: 40% EtOH (0.05% DEA) under CO2;

[0294] Flow rate: 2.5 mL / min;

[0295] ABPR: 1500psi;

[0296] Temperature: 35℃.

[0297] The compound with a retention time of 2.903 min was defined as isomer 1; MS (ESI) m / z 418.3 [M+H] +

[0298] The compound with a retention time of 3.997 min was defined as isomer 2; MS (ESI) m / z 418.3 [M+H] +

[0299] Example 2

[0300] (3R,4R)-4-((5-chloro-4-(8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazaacenaphthyl-6-yl)pyrimidin-2-yl)amino)-1-(methylsulfonyl)piperidin-3-ol Isomer 1

[0301] (3R,4R)-4-((5-chloro-4-(8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazaacenaphthyl-6-yl)pyrimidin-2-yl)amino)-1-(methylsulfonyl)piperidin-3-ol Isomer 2

[0302]

[0303] first step

[0304] 6-(2,5-Dichloropyrimidin-4-yl)-8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylene 2a

[0305] Under a nitrogen atmosphere, compound 1f (50 mg, 0.18 mmol), pinacol diboron (67 mg, 0.26 mmol), potassium acetate (37 mg, 0.38 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (26 mg, 0.04 mmol) were dissolved in 2 mL of 1,4-dioxane. The mixture was reacted at 100°C for 1 hour. After cooling to room temperature, 1,3,5,7-tetramethyl-6-phenyl-2,4,8-trioxa-6-phosphoryladamantane (10 mg, 0.04 mmol), potassium carbonate (51 mg, 0.37 mmol), 2,4-dichloro-5-fluoropyrimidine (52 mg, 0.28 mmol), tris(dibenzylideneacetone)dipalladium (32 mg, 0.04 mmol), and 0.5 mL of water were added and the reaction was continued at 80°C for 1 hour. The reaction solution was cooled to room temperature, filtered, and the filtrate was collected and concentrated under reduced pressure. The residue was purified by C-18 reverse phase chromatography to give the title compound 2a (30 mg, yield: 48%).

[0306] MS (ESI) m / z 353.1 [M+H] +

[0307] Step 2

[0308] (3R,4R)-4-((5-chloro-4-(8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazaacenaphthyl-6-yl)pyrimidin-2-yl)amino)-1-(methylsulfonyl)piperidin-3-ol Isomer 1

[0309] (3R,4R)-4-((5-chloro-4-(8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazaacenaphthyl-6-yl)pyrimidin-2-yl)amino)-1-(methylsulfonyl)piperidin-3-ol Isomer 2

[0310] Under a nitrogen atmosphere, compound 2a (30 mg, 0.08 mmol), (3R,4R)-4-amino-1-(methylsulfonyl)piperidin-3-ol (19.4 mg, 0.10 mmol, prepared using the method disclosed in patent application "WO 2019 / 207463 A1"), (S)-(-)-2,2"-bis(diphenylphosphino)-1,1"-binaphthyl (10 mg, 0.02 mmol), and palladium acetate (4.4 mg, 0.02 mmol) were dissolved in 2 mL of tetrahydrofuran. Cesium carbonate (52 mg, 0.16 mmol) was added, and the reaction was carried out at 80°C for 1 hour. The reaction solution was cooled to room temperature, filtered, and the filtrate was collected. The residue was concentrated under reduced pressure and purified by C-18 reverse phase chromatography to give a crude mixture. The crude product was subjected to chiral separation (column: DAICEL CHIRALCEL OD-H (250 mm*30 mm, 5 μm), 50% EtOH (0.1% NH 3 · H 2 O) in CO 2 ; flow rate: 80 mL / min) to give isomer 1 (4.4 mg, yield: 10%) and isomer 2 (3.3 mg, yield: 8%).

[0311] Analytical methods

[0312] Column: DAICEL CHIRALPAK AD-3 (50 mm*4.6 mm, 3 μm);

[0313] Mobile phase: A: CO2 B: isopropanol (0.05% DEA), gradient: 5% to 40% B in 2 min, maintain 40% B for 1.2 min, then 5% B for 0.8 min;

[0314] Flow rate: 4 mL / min;

[0315] ABPR:1500psi;

[0316] Temperature: 35℃.

[0317] The compound with a retention time of 2.094 min was defined as isomer 1;

[0318] MS (ESI) m / z 511.3 [M+H] +

[0319] 1H NMR (400MHz, DMSO-d6) δ=8.38(s,1H),7.48(br s,1H),6.91(br d,J=11.0Hz,1H),5.21(br s,1H),4.86(br d,J=6.5Hz,1H),4.52-4.44(m,1H),4.24(br d,J=10.8Hz,1H),3.77(br s,1H),3.64-3.55(m,2H),3.48(br d,J=13.6Hz,1H),2.89(s,3H),2.69-2.63(m,1H),2.60(s,3H),1.39(d,J=6.8Hz,3H),1.24(br s, 2H), 1.18-1.04 (m, 1H).

[0320] The compound with a retention time of 2.499 min was defined as isomer 2;

[0321] MS (ESI) m / z 511.3 [M+H] +

[0322] 1 H NMR (400MHz, DMSO-d6) δ = 8.38 (s, 1H), 7.47 (br s, 1H), 6.91 (br d, J = 11.5Hz, 1H), 5.21 (br d, J = 4.0Hz, 1H), 4.86 (br d,J=6.5Hz,1H),4.48(dd,J=1.8,11.5Hz,1H),4.36(t,J=5.0Hz,2H),4.24(br d,J=9.5Hz,1H),3.76(br s,1H),3.59(brd,J=8.0Hz,2H),2.89(s,3H),2.67(br d,J=9.3Hz,1H),2.60(s,3H),1.58-1.46(m,1H),1.39(d,J=6.8Hz,3H),1.24(br s,1H).

[0323] Example 3

[0324] (3S,4R)-4-((5-chloro-4-((S)-8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylene-6-

[0325] (4-(2-Yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol Isomer 1)

[0326] (3S,4R)-4-((5-chloro-4-((R)-8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylene-6-

[0327] 1-[(1-amino-2-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol isomer 2]

[0328]

[0329] Under a nitrogen atmosphere, compound 2a (55 mg, 0.16 mmol), (3S, 4R)-4-aminotetrahydro-2H-pyran-3-ol (18 mg, 0.16 mmol), (S)-(-)-2,2"-bis(diphenylphosphino)-1,1"-binaphthyl (20 mg, 0.03 mmol), and palladium acetate (6.7 mg, 0.03 mmol) were dissolved in 3 mL of tetrahydrofuran. Cesium carbonate (104 mg, 0.32 mmol) was added and the reaction was carried out at 85°C for 3 hours. The reaction solution was cooled to room temperature, filtered, and the filtrate was collected. After concentration under reduced pressure, the residue was purified by C-18 reverse phase chromatography to obtain a crude mixture. The crude product was separated by chiral separation [column: DAICEL CHIRALPAK AD (250 mm*30 mm, 10 μm), conditions: 0.1% NH3 . H2O IPA, starting with B: 45%; ending with B: 45%; flow rate (ml / min): 70)] to give Isomer 1 (6.2 mg, yield: 9.2%) and Isomer 2 (7.7 mg, yield: 11%).

[0330] Analytical methods

[0331] Column: DAICEL CHIRALCELOD-3 (100 mm*4.6 mm, 3 μm);

[0332] Mobile phase: A: CO2 B: ethanol (0.05% DEA), gradient: 5% to 40% B in 4 min, maintain 40% B for 2.5 min, then 5% B for 1.5 min;

[0333] Flow rate: 2.8 mL / min;

[0334] ABPR:1500psi;

[0335] Temperature: 35℃.

[0336] The compound with a retention time of 3.518 min was defined as isomer 1;

[0337] MS (ESI) m / z 434.3 [M+H] +

[0338] 1 H NMR (400MHz, DMSO-d6) δ = 8.37 (s, 1H), 7.45 (br s, 1H), 6.90 (d, J = 11.5Hz, 1H), 4.93 (d, J = 5.3Hz, 1H), 4.86 (br d,J=6.8Hz,1H),4.48(dd,J=1.6,11.7Hz,1H),4.24(br d,J=9.3Hz,1H),3.84-3.74(m,3H),3.53-3.40(m,2H),3.03(br t,J=10.2Hz,1H),2.60(s,3H),2.01-1.90(m,1H),1.54-1.44(m,1H),1.38(d,J=6.5Hz,3H).

[0339] The compound with a retention time of 4.165 min was defined as isomer 2;

[0340] MS (ESI) m / z 434.3 [M+H] +

[0341] 1 H NMR (400MHz, DMSO-d6) δ=8.37(s,1H),7.46(br s,1H),6.91(br d,J=11.5Hz,1H),4.93(d,J=5.3Hz,1H),4.86(br d,J=6.5Hz,1H),4.47(br d,J=10.5Hz,1H),4.23(br d,J=9.8Hz,1H),3.86-3.74(m,3H),3.54-3.44(m,1H),3.32-3.27(m,1H),3.03(br t,J=10.3Hz,1H),2.60(s,3H),1.95(br d,J=13.1Hz,1H),1.54-1.43(m,1H),1.38(d,J=6.5Hz,3H).

[0342] Example 4

[0343] (3S,4R)-4-((5-chloro-4-((S)-8-fluoro-2-(2-hydroxypropan-2-yl)-3-methyl-3,4-dihydro-5-oxapropane)

[0344] -1,2a-diazaacenaphthylene-6-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol isomer 1

[0345] (3S,4R)-4-((5-chloro-4-((R)-8-fluoro-2-(2-hydroxypropan-2-yl)-3-methyl-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylene-6-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol Isomer 2

[0346]

[0347]

[0348] first step

[0349] 6-Bromo-8-fluoro-3-methyl-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylene-2-carbaldehyde 4a

[0350] Under a nitrogen atmosphere, compound 1f (2.0 g, 7 mmol) and selenium dioxide (3.1 g, 28 mmol) were added sequentially to 30 mL of 1,4-dioxane. The reaction was allowed to proceed at 95°C for 8 hours. The reaction solution was cooled to room temperature, filtered, and the filtrate was collected and concentrated under reduced pressure. The residue was purified by C-18 reverse-phase chromatography to afford the title compound 4a (930 mg, 44% yield).

[0351] MS (ESI) m / z 299.1, 301.1 [M+H] +

[0352] Step 2

[0353] 1-(6-Bromo-8-fluoro-3-methyl-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylene-2-yl)ethan-1-ol 4b

[0354] Under a nitrogen atmosphere, compound 4a (930 mg, 3.1 mmol) was dissolved in 20 mL of tetrahydrofuran. The temperature was cooled to -20°C, and a solution of methylmagnesium bromide in tetrahydrofuran (3 mol / L, 1.5 mL, 4.5 mmol) was added dropwise. The mixture was allowed to react at -20°C for 4 hours. The reaction was quenched by the addition of 5 mL of water. The reaction solution was concentrated under reduced pressure, and the residue was purified by C-18 reverse-phase chromatography to afford the title compound 4b (830 mg, 85% yield).

[0355] MS (ESI) m / z 315.2, 317.2 [M+H] +

[0356] Step 3

[0357] 1-(6-Bromo-8-fluoro-3-methyl-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylene-2-yl)ethan-1-one 4c

[0358] At room temperature, compound 4b (600 mg, 1.9 mmol) was dissolved in 20 mL of tetrahydrofuran, and Dess-Martin periodinane (2.0 g, 4.8 mmol) was added. The mixture was heated to 80°C and reacted for 2 hours. The reaction solution was cooled to room temperature, filtered, and the filtrate was collected. The residue was concentrated under reduced pressure and purified by C-18 reverse phase chromatography to obtain the title compound 4c (350 mg, yield: 59%).

[0359] MS (ESI) m / z 313.1, 315.1 [M+H] +

[0360] Step 4

[0361] 2-(6-Bromo-8-fluoro-3-methyl-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylene-2-yl)propan-2-ol 4d

[0362] Under a nitrogen atmosphere, compound 4c (350 mg, 1.1 mmol) was dissolved in 20 mL of tetrahydrofuran. The temperature was cooled to -20°C, and a solution of methylmagnesium bromide in tetrahydrofuran (3 mol / L, 0.7 mL, 2.1 mmol) was added dropwise. The mixture was allowed to react at -20°C for 4 hours. The reaction was quenched by the addition of 5 mL of water. The reaction solution was concentrated under reduced pressure, and the residue was purified by C-18 reverse-phase chromatography to afford the title compound 4d (260 mg, 71% yield).

[0363] MS (ESI) m / z 329.2, 331.2 [M+H] +

[0364] Step 5

[0365] 2-(6-(2,5-dichloropyrimidin-4-yl)-8-fluoro-3-methyl-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylene-2-yl)propane

[0366] -2-ol 4e

[0367] Under a nitrogen atmosphere, compound 4d (260 mg, 0.8 mmol), pinacol diboronate (305 mg, 1.2 mmol), potassium acetate (157 mg, 1.6 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (117 mg, 0.2 mmol) were dissolved in 5 mL of 1,4-dioxane. The mixture was reacted at 100°C for 2 hours. After cooling to room temperature, 1,3,5,7-tetramethyl-6-phenyl-2,4,8-trioxa-6-phosphoryladamantane (58 mg, 0.2 mmol), potassium carbonate (221 mg, 1.6 mmol), 2,4-dichloro-5-fluoropyrimidine (220 mg, 1.2 mmol), tris(dibenzylideneacetone)dipalladium (183 mg, 0.2 mmol), and 1 mL of water were added and the reaction was continued at 80°C for 1 hour. The reaction solution was cooled to room temperature, filtered, and the filtrate was collected and concentrated under reduced pressure. The residue was purified by C-18 reverse phase chromatography to give the title compound 4e (129 mg, yield: 41%).

[0368] MS (ESI) m / z 397.3 [M+H] +

[0369] Step 6

[0370] (3S,4R)-4-((5-chloro-4-((S)-8-fluoro-2-(2-hydroxypropan-2-yl)-3-methyl-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylene-6-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol Isomer 1

[0371] (3S,4R)-4-((5-chloro-4-((R)-8-fluoro-2-(2-hydroxypropan-2-yl)-3-methyl-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylene-6-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol Isomer 2

[0372] Under a nitrogen atmosphere, compound 4e (124 mg, 0.31 mmol), (3S,4R)-4-aminotetrahydro-2H-pyran-3-ol (19 mg, 0.16 mmol), (S)-(-)-2,2"-bis(diphenylphosphino)-1,1"-binaphthyl (118 mg, 0.5 mmol), and palladium acetate (14 mg, 0.06 mmol) were dissolved in 5 mL of tetrahydrofuran. Cesium carbonate (202 mg, 0.62 mmol) was added, and the mixture was reacted at 85°C for 3 hours. The reaction solution was cooled to room temperature, filtered, and the filtrate was collected. The residue was concentrated under reduced pressure and purified by C-18 reverse phase chromatography to obtain the crude product 4f. The crude product was separated by chiral separation [column: DAICEL CHIRALPAK AD (250 mm*30 mm, 10 μm), conditions: 0.1% NH3. H2OETOH, Begin B: 40%; End B: 40%; flow rate (ml / min): 60)] to obtain the title compound isomer 1 (25.2 mg, yield: 17%) and the title compound isomer 2 (22 mg, yield: 15%).

[0373] Analytical methods

[0374] Column: DAICEL CHIRALCEL AD-3 (100 mm*4.6 mm, 3 μm);

[0375] Mobile phase: A: CO2 B: ethanol (0.05% DEA), gradient: 5% to 40% B in 2 min, maintain 40% B for 1.2 min, then 5% B for 0.8 min;

[0376] Flow rate: 4 mL / min;

[0377] ABPR:1500psi;

[0378] Temperature: 35℃.

[0379] The compound with a retention time of 1.887 min was defined as isomer 2;

[0380] MS (ESI) m / z 478.1 [M+H] +

[0381] 1 H NMR (400MHz, DMSO-d6) δ = 8.37 (s, 1H), 7.45 (br s, 1H), 6.92 (d, J = 11.3Hz, 1H), 5.82 (br s,1H),5.23(q,J=6.4Hz,1H),4.93(d,J=5.3Hz,1H),4.47(d,J=11.0Hz,1H),4.22(br d,J=11.3Hz,1H),3.89-3.67(m,3H),3.39-3.25(m,2H),3.03(br t,J=10.2Hz,1H),1.95(br d,J=10.5Hz,1H),1.68(s,3H),1.62(s,3H),1.54-1.47(m,1H),1.45(d,J=6.5Hz,3H).

[0382] The compound with a retention time of 2.078 min was defined as isomer 1;

[0383] MS (ESI) m / z 478.1 [M+H] +

[0384] 1H NMR (400MHz, DMSO-d6) δ = 8.37 (s, 1H), 7.45 (br s,1H),6.92(d,J=11.5Hz,1H),5.82(s,1H),5.29-5.19(m,1H),4.93(d,J=5.5Hz,1H),4.47(d,J=11.3Hz,1H),4.21(br d,J=10.3Hz,1H),3.85-3.75(m,3H),3.53-3.40(m,2H),3.03(br t,J=10.4Hz,1H),1.94(br s,1H),1.67(s,3H),1.62(s,3H),1.53-1.47(m,1H),1.45(d,J=6.5Hz,3H).

[0385] Biological evaluation

[0386] The present invention is further described and explained below in conjunction with test examples, but these examples are not intended to limit the scope of the present invention.

[0387] Test Example 1: Detection of Cyclin-Dependent Kinase Activity of Disclosed Compounds

[0388] 1. Experimental Materials

[0389]

[0390] Compound A is Example A94 of WO 2019 / 207463A1, and was synthesized according to the method disclosed in the patent application.

[0391] 2. Kinase activity test (CDK4 / cyclin D1, CDK6 / cyclin D3)

[0392] In vitro CDK kinase activity was assessed using a mobility shift assay. Inhibition of CDK activity by the test compound was assessed at a starting concentration of 300 nM, using 3-fold dilutions for a total of 10 concentrations in duplicate. Staurosporine was used as a standard control.

[0393] Prepare 1x kinase buffer (CDK2) (50 mM HEPES, pH 7.5, 0.0015% Brij-35), 1x kinase buffer (CDK4) (20 mM HEPES, pH 7.5, 0.01% Triton X-100), and stop solution (100 mM HEPES, pH 7.5, 0.015% Brij-35, 0.2% Coating Reagent #3, 50 mM EDTA). Add the appropriate amount of kinase to the 1x kinase buffer to prepare a 2.5x enzyme solution. Prepare a 5x compound dilution solution (1x kinase buffer, 10% DMSO) corresponding to the test compound concentration. Add the appropriate amount of FAM-labeled peptide and ATP to the 1x kinase buffer to prepare a 2.5x substrate solution. 5 μl of 5x compound dilution and 10 μl of 2.5x enzyme solution were added to the reaction wells of a 384-well reaction plate, mixed, and incubated at room temperature for 10 minutes. 10 μl of 2.5x substrate solution was then added to the 384-well plate, and the plate was centrifuged at 1000 rpm for 1 minute. The plate was incubated at 28°C for 60 minutes (biochemical incubator model: SPX-100B-Z). 30 μl of stop solution was added to the 384-well plate to terminate the reaction, and the plate was centrifuged at 1000 rpm for 1 minute. Conversion rate data were read on a Caliper EZ Reader II (excitation wavelength: 400 nm, emission wavelengths: 445 nm and 520 nm).

[0394] IC of the compound 50 The values ​​were fitted using XLFit Excel add-in version 5.4.0.8. Fitting formula:

[0395] Y=Bottom+(Top-Bottom) / (1+10^((LogIC50-X)*HillSlope)).

[0396] X: log value of compound concentration; Y: percentage of compound inhibition.

[0397] 3. Kinase activity test (CDK1 / cyclin B, CDK9 / cyclin T1)

[0398] In vitro CDK (CDK2, CDK9) kinase activity assays were performed with a starting concentration of 1 μM, followed by 3-fold dilutions for a total of 10 concentrations in duplicate wells. PHA-793887 was used as a control compound.

[0399] Prepare 1x kinase reaction buffer (40mM Tris-HCl, pH 7.4, 20mM MgCl2, 0.1mg / ml BSA, 50μM DTT) by adding 1 volume of 5x kinase reaction buffer and 4 volumes of water. Add DTT (final concentration 50μM). Use an Echo655 to transfer 50nL of the diluted compound working solution (final DMSO concentration 1%) to each well of a reaction plate (784075, Greiner). Seal the plate with film and centrifuge at 1000g for 1 minute. Prepare 2x enzyme (0.3ng / uL CDK2 / cyclin E1 or CDK9 / cyclin T1) in 1x kinase reaction buffer. Add 2.5uL of the above kinase solution to each well. Seal the plate with film and centrifuge at 1000g for 1 minute. Incubate at room temperature for 10 minutes. Prepare a 2× kinase substrate and ATP mixture in 1× kinase reaction buffer. The 2× CDK2 / CylinE1 kinase substrate consists of 0.4 mg / mL histone H1 and 30 μM ATP. Add 2.5 μL of the 2× histone H1 and ATP mixture to the reaction plate and centrifuge at 1000 g for 30 seconds to initiate the reaction. After the kinase assay is incubated at room temperature for 120 minutes, add 4 μL of ADP-Glo ​​reagent and incubate at room temperature for 40 minutes. Then, add 8 μL of kinase detection reagent and incubate at room temperature for 40 minutes. Luminescence signals are read using an Envision 2104.

[0400] The data analysis is as follows

[0401] a) Inhibition percentage: % inhibition

[0402] =100-(Signalcmpd-SignalAve_PC) / (SignalAve_VC-SignalAve_PC)×100.

[0403] SignalAve_PC: The average value of all positive control wells in the whole plate.

[0404] SignalAve_VC: The average value of all negative control wells in the whole plate.

[0405] Signalcmpd: The average value of the corresponding wells of the test compound.

[0406] b) Compound IC 50 : Calculated using GraphPad 8.0 using the following nonlinear fitting formula.

[0407] Y=Bottom+(Top-Bottom) / (1+10^((LogIC50-X)*HillSlope))

[0408] X: log value of compound concentration; Y: percentage of compound inhibition.

[0409] The CDK (CDK1, CDK4, CDK6, CDK9) kinase biochemical inhibitory activity of the disclosed compounds was determined by the above assay, and the measured IC 50 The values ​​are shown in Tables 1, 2 and 3.

[0410] Table 1.

[0411]

[0412] Table 2.

[0413]

[0414] Table 3.

[0415]

[0416]

[0417] Test Example 2: CYP inhibition experiment

[0418] Pooled human liver microsomes from 150 donors (Corning, Cat. No. 452117) were used to evaluate the metabolic reactions of representative substrates of the five major human CYP isoforms (CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4). The effects of varying concentrations of the test compounds on the metabolic reactions of phenacetin (CYP1A2), diclofenac sodium (CYP2C9), S-mephenytoin (CYP2C19), bufuralol hydrochloride (CYP2D6), and midazolam (CYP3A4 / 5) were determined by liquid chromatography-tandem mass spectrometry (LC / MS / MS).

[0419] A 200 μL reaction mixture (100 mmol / L phosphate buffer, pH 7.4, containing 0.3% DMSO, 0.6% acetonitrile, and 0.1% methanol, by volume) containing 30 μM phenacetin, 10 μM diclofenac sodium, 35 μM S-mephenytoin, 5 μM bufuralol hydrochloride, 3 μM midazolam, 1 mM NADPH, test compounds (at concentrations of 0.1, 0.3, 1, 3, 10, and 30 μmol / L), positive compounds, or a blank control, was incubated with mixed human liver microsomes (0.2 mg / mL) for 5 minutes at 37°C. Then, 200 μL of an acetonitrile solution containing 3% formic acid and 40 nM internal standard verapamil was added, and the mixture was centrifuged at 4000 rpm for 50 minutes. The mixture was cooled on ice for 20 minutes and then centrifuged at 4000 rpm for 20 minutes to precipitate the protein. A 200 μL supernatant was used for LC / MS / MS analysis.

[0420] The peak area was calculated based on the chromatogram. The residual activity ratio (%) was calculated using the following formula:

[0421] Peak area ratio = metabolite peak area / internal standard peak area

[0422] Residual activity ratio (%) = peak area ratio of the test compound group / peak area ratio of the blank group

[0423] CYP half-maximal inhibitory concentration (IC 50 ) was calculated using Excel XLfit 5.3.1.3.

[0424] The measured CYP half-maximal inhibitory concentration (IC 50 ) values ​​are shown in Table 4 below.

[0425] Table 4. The half-maximal inhibitory concentration (IC) of the compounds of the present disclosure on CYP 50 )

[0426]

[0427] Test Example 3: Solubility Test

[0428] The thermodynamic solubility of the compounds was determined in phosphate buffer at pH 7.4. Sample supernatants and standards of known concentrations were analyzed by LC / MS / MS.

[0429] 1. Materials and Reagents

[0430] Compound A (Compound A is Example A94 of WO 2019207463A, synthesized according to the method disclosed in the patent application).

[0431] NaH2PO4·2H2O (analytical grade), NaH2PO4 (analytical grade), NaOH (analytical grade).

[0432] 1.5 mL flat-bottom glass tubes (BioTech Solutions); molded polytetrafluoroethylene caps (BioTech Solutions), polytetrafluoroethylene-coated stirring bars (BioTech Solutions), Eppendorf Comfort Thermomixer, and 96-well deep-well plates.

[0433] 2. Preparation of 0.01 M sodium phosphate buffer at pH 7.4

[0434] Weigh 15.6 g of NaH₂PO₄·2H₂O into a 1 L glass bottle and dissolve in 1 L of deionized water. The pH of the solution should be approximately 4.7. Adjust the pH to 7.4 with 10 M NaOH.

[0435] 3. Solubility determination process

[0436] Accurately weigh 1 mg of powder of each compound and place it into a glass tube. Add 1 mL of pH 7.4 phosphate buffer per mg to the tube. Add a stir bar to each tube, close the lid, and place the sample plate containing the tubes in an Eppendorf Comfort Thermomixer. Incubate at 25°C, 1100 rpm for 24 hours. After the incubation period, open the lid, remove the stir bar using a magnet, and record the contents of each tube. Centrifuge the plate at 25°C, 4000 rpm for 30 minutes. Aspirate 750 μL of the supernatant. Rinse the pipette tip with acetonitrile for 5 seconds, then with pure water for 5 seconds. The first 50 μL of the solution was then discarded, and the remaining 700 μL was added to another 96-well sample plate containing glass tubes. The sample was centrifuged again for 30 minutes (25°C, 4000 rpm). 10 μL of the sample from the second centrifugation was added to 990 μL of a 1:1 mixture of acetonitrile and water containing the internal standard (100x sample). Furthermore, 10 μL of the dilution solution was added to 990 μL of a 1:1 mixture of acetonitrile and water containing the internal standard (10,000x sample). The sample dilution factor may vary depending on the solubility value and LC / MS signal response.

[0437] Table 5. Phenomenon records and dilution factors

[0438]

[0439] 4. Preparation of Standards

[0440] Accurately weigh 1 mg of compound powder and add it to a glass tube. Add 1 mL of DMSO per mg to each tube. Add a stir bar to each tube and cap. Place the tray containing the standard tubes in an Eppendorf Comfort Thermomixer and incubate at 1,100 rpm at 25°C for 2 hours to allow the powder to fully dissolve. Observe for complete dissolution of the solids and note any compounds that do not dissolve completely in the DMSO solution. Pipette 10 μL of the 1 mg / mL standard into 990 μL of a 1:1 mixture of acetonitrile and water containing the internal standard to obtain a 10 μg / mL standard. Pipette 10 μL of the 10 μg / mL standard into 990 μL of a 1:1 mixture of acetonitrile and water containing the internal standard to obtain a 0.1 μg / mL standard. Sample dilution may vary depending on LC / MS signal response. Samples were analyzed by LC / MS / MS. All compounds were tested individually.

[0441] 5. Data calculation

[0442] All calculations were performed using Microsoft Excel.

[0443] Samples were analyzed by LC / MS / MS and quantified based on standards of known concentrations. The solubility of the test compound was calculated using the following formula: [Sample] = Area Ratio Sample × DF Sample × [STD] / Area Ratio STD DF: dilution factor.

[0444] STD: Standard of the compound to be tested.

[0445] Table 6. Solubility of compounds of the present disclosure

[0446] Example No. Solubility (μM) Compound A 16 Isomer 2 with a retention time of 1.887 min in Example 4 2126 Isomer 1 with a retention time of 2.078 min in Example 4 2203

[0447] Test Example 4: PXR Induction Experiment

[0448] 1. Evaluate the potential of the test compound to induce drug metabolizing enzyme activity by activating PXR in vitro. The test compound was measured at different concentrations (30, 10, 3.33, 1.11, 0.370 and 0.123 μM) to obtain EC 50 The concentrations of rifampicin as the positive control were 20, 5, 1.25, 0.312, 0.0781, and 0.195 μM.

[0449] 2. Materials and Reagents

[0450] 1) DPX2 (human PXR gene and fluorescent reporter gene stably transfected into HepG2 cells) cells were purchased from Puracyp (Carlsbad, CA).

[0451] 2) The client provided the test compound, and the control drug (rifampicin) was purchased from Sigma (St. Louis, MO).

[0452] 3) CellTiter-Fluor TM The cell viability assay kit and One-Glo fluorescence assay kit were purchased from Promega (Madison, WI), fetal bovine serum (FBS) was purchased from Corning (Manassas, VA), and an MTS3 shaker was purchased from IKA Labortechnik (Staufen, Germany). DMEM, penicillin, and streptomycin were purchased from local suppliers, hygromycin B and G418 were purchased from Merck (Darmstadt, Germany), and cell culture medium and DPX2 cells were purchased from Puracyp Inc.

[0453] 3. Experimental steps

[0454] 3.1 Seed Plate Preparation

[0455] 1) Add 50 mL of FBS to 450 mL of cell culture medium.

[0456] 2) DPX2 cells were cultured in a T-75 culture flask in an incubator at 37° C., 5% CO 2 , and 95% relative humidity. The cells were digested when the cells covered 80-90% of the bottom of the culture flask.

[0457] 3) Wash the surface of the T-75 cultured cells with 8 mL of PBS, discard the PBS, add 3 mL of trypsin, and digest at 37°C for about 5 minutes, or until the cells are digested and suspended in trypsin. Add 10 mL of excess serum-containing culture medium to neutralize the trypsin.

[0458] 4) Transfer the cell suspension to a conical-bottom centrifuge tube and centrifuge at 120 g for 10 minutes. Resuspend the cells in seed culture medium and adjust the concentration to 4 x 10 5 cells / mL. Add 100 μL of diluted cells to each well of a 96-well cell culture plate. Place the plate in an incubator at 37°C for 24 hours and then prepare for the PXR activation assay.

[0459] 3.2 Dosing treatment

[0460] 1) Prepare the test compound and positive compound (rifampicin) in DMSO and dilute the compound in serum-free medium at 37°C. Final concentrations of rifampicin as the positive control are 20, 5, 1.25, 0.312, 0.0781, and 0.195 μM, and final concentrations of the test compound are 30, 10, 3.33, 1.11, 0.370, and 0.123 μM. The final DMSO concentration is 0.1%. Add 1 μL of DMSO to 1 mL of preincubated medium as a solvent control.

[0461] 2) Take the cell culture plate out of the incubator, discard the culture medium, add 100 μL of the test compound and the positive compound to the appropriate wells, two parallel wells per group, and place the cell plate in the incubator for 24 hours.

[0462] Quantitative determination of PXR activation

[0463] 1) Two days after drug treatment, cultures can be used for quantitative detection of PXR activation.

[0464] 2) CellTiter-Fluor TM Equilibrate the Cell Viability Assay Kit and One-Glo Luciferase Reagent to room temperature. Add GF-AFC substrate (10 μL) to assay buffer (10 mL) to form a 2X reagent, then dilute to 1X with 10 mL of PBS. Add ONE-Glo substrate to ONE-Glo Luciferase Assay buffer.

[0465] 3) Remove the cell culture plate from the incubator, discard the culture medium in each well, wash twice with PBS, and add 1X CellTiter-FluorTM The reagent was added to the sterilized sample reservoir, 50 μL was drawn into each well using a dispenser, and incubated at 37°C for 30 minutes.

[0466] 4) Remove the 96-well cell plate from the incubator and measure the fluorescence value of each well using a microplate reader in fluorescence mode with an excitation wavelength of 400 nm and an emission wavelength of 505 nm.

[0467] 5) Pour ONE-Glo reagent into the sample reservoir, then use a dispenser to dispense 50 μL into each well. Gently mix the reagent and incubate at room temperature for 5 minutes. After incubation, read the luminescence value of each well using a luminometer.

[0468] 4. Calculation of cell induction value

[0469] 4.1 Cell viability

[0470] Cell viability calculation formula:

[0471] Cell viability percentage (%) = I (sample) / (I (vehicle) x 100

[0472] I(sample) is the fluorescence intensity of the sample, and I(vehicle) refers to the fluorescence intensity of the cells in the presence of 0.1% DMSO.

[0473] 4.2 Calculation of cell induction value

[0474] All data were calculated using Microsoft Excel.

[0475] The activity of luciferase is expressed as RFU / RLU, where RLU is the average luminescence intensity of two parallel assays for each compound at each concentration, and RFU is the average fluorescence intensity of two parallel assays for each compound at each concentration. The induction multiple is calculated as follows:

[0476]

[0477] Table 7. Some of the measured PXR induction values

[0478]

Claims

1. A pharmaceutical composition comprising 0.01-99.99% of a compound of formula I or a pharmaceutically acceptable salt thereof, based on the total weight of the composition, and a pharmaceutically acceptable excipient. in, R 1 Selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl and C 3-8 Cycloalkyl, the C 1-6 Alkyl, C 1-6 Haloalkyl and C 3-8 The cycloalkyl groups are each independently optionally substituted with one or more R a replace; R 2 The structure of formula II: R 9 Selected from H, OH and NH2, wherein NH2 is optionally replaced by 1 or 2 R a’ or R a” replace; R 10 Each independently selected from OH, halogen, CN, NH2, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 3-8 The cycloalkyl group and the 3-12 membered heterocycloalkyl group are each independently optionally substituted with one or more R b replace; Q is NR 11 or O; Or Q is CR 12 R 13 , where R 12 、R 13 The carbon atom connected to it forms a 3-12-membered NR 11 wherein N or O is a heterocycloalkyl group as a ring atom, the heterocycloalkyl group is optionally substituted by one or more R 10 replace; R 11 Selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, SO2R c 、SO2NR d R e 、COR f 、COOR f and CONR g R h , the C 1-6 Alkyl and C 1-6 Each haloalkyl group is independently optionally selected from R a 、R b 、SO2R c 、SO2NR d R e 、COR f 、COOR f and CONR g R h substituted with one or more substituents; m is 0, 1, or 2; n is 0, 1, 2, 3 or 4; p is 1, 2, or 3; X is N or CH; Y is N or CR 7 ; R 7 Selected from H, F, Cl, CN, C 1-6 Alkyl and C 1-6 Alkoxy, the C 1-6 Alkyl and C 1-6 Each alkoxy group is independently optionally substituted with one or more R a replace; R 3 selected from H, F, Cl, CN, CH2F, CHF2 and CF3; R 4 Selected from H, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 The cycloalkyl group and the 3-12 membered heterocycloalkyl group are each independently optionally substituted with one or more R b or deuterium substitution; Z is O or CHR 8 , R 8 selected from the group consisting of hydrogen atoms, deuterium atoms and halogens; L is -(CH2) q -, wherein -(CH2)- is optionally selected from deuterium, CN, halogen, C 1-6 Alkyl, C 3-8 Cycloalkyl, C 1-6 Alkoxy and 3-12 membered heterocycloalkyl are substituted with one or more substituents, the C 1-6 Alkyl, C 3-8 Cycloalkyl, C 1-6 Alkoxy and 3-12 membered heterocycloalkyl are each independently optionally substituted with one or more R b or deuterium substitution; q is 1, 2, 3, or 4; R 5 and R 6 Each independently selected from H, deuterium, CN, halogen, C 1-6 Alkyl, C 3-8 Cycloalkyl, C 1-6 Alkoxy and 3-12 membered heterocycloalkyl, the C 1-6 Alkyl, C 3-8 Cycloalkyl, C 1-6 Alkoxy and 3-12 membered heterocycloalkyl are each independently optionally substituted with one or more R b or deuterium substitution; R a and R b Each independently selected from H, OH, CN, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-8 Cycloalkyl, 3-12 membered heterocycloalkyl and NR a’ R a” , the C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl are each independently optionally selected from NH2, NHCH3, N(CH3)2, halogen, OH, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl are substituted with one or more substituents; R a’ and R a” Each independently selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl, the C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl are each independently optionally selected from NH2, NHCH3, N(CH3)2, halogen, OH, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl are substituted with one or more substituents; or R a’ 、R a” Together with the nitrogen atom to which they are attached, they form a 3-12 membered heterocycloalkyl group, wherein the 3-12 membered heterocycloalkyl group is optionally selected from halogen, OH, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl are substituted with one or more substituents; R c 、R d and R e Each independently selected from H, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 Cycloalkyl and C 1-6 alkyl halide; R f Selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl, the C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl are each independently optionally selected from NH2, NHCH3, N(CH3)2, halogen, OH, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl are substituted with one or more substituents; R g and R h Each independently selected from H, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl and C 3-8 Cycloalkyl, the C 1-6 Alkoxy, C 1-6 Haloalkyl and C 3-8 The cycloalkyl groups are each independently optionally substituted with one or more R a or R b replace.

2. The pharmaceutical composition according to claim 1, wherein R 1 H or C 1-6 Alkyl, preferably H.

3. The pharmaceutical composition according to claim 1, wherein R 2 Selected from Preferred R 9 、R 10 , m and Q are as defined in claim 1.

4. The pharmaceutical composition according to claim 3, wherein R 2 for R 9 、R 10 , m and R 11 As defined in claim 1.

5. The pharmaceutical composition according to claim 3 or 4, wherein R 9 is OH or NH2, wherein the NH2 is optionally replaced by 1 or 2 R a’ or R a” Replacement, R a’ and R a” Each independently is C 1-6 alkyl. The pharmaceutical composition according to claim 1 , wherein X is N.

7. The pharmaceutical composition according to claim 1, wherein Y is CR 7 ; R 7 Selected from H, F, Cl and C 1-6 Alkyl, preferably F or Cl.

8. The pharmaceutical composition according to claim 1, wherein R 3 is selected from H, F and Cl, preferably F or Cl.

9. The pharmaceutical composition according to claim 1, wherein R 4 Selected from H, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 The cycloalkyl group and the 3-12 membered heterocycloalkyl group are each independently optionally substituted with one or more R b or deuterium substitution, R b Selected from OH, CN, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Haloalkoxy and C 3-8 Cycloalkyl; preferably, R 4 C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with one or more R b or deuterium substitution, R b is OH or halogen.

10. The pharmaceutical composition according to claim 1, wherein L is -(CH2) q -, q is selected from 1 or 2; said -(CH2)- is optionally selected from deuterium, CN, halogen, C 1-6 Alkyl, C 3-8 Cycloalkyl, C 1-6 Alkoxy and one or more substituents of 3-12 membered heterocycloalkyl; Preferably, q is selected from 1; said -(CH2)- is optionally selected from H, deuterium, CN, halogen, C 1-6 Alkyl, C 3-8 Cycloalkyl, C 1-6 Alkoxy and one or more substituents of 3-12 membered heterocycloalkyl; Most preferably, q is selected from 1; and said -(CH2)- is optionally substituted with one or more deuterium.

11. The pharmaceutical composition according to claim 1, wherein R 5 and R 6 Each independently selected from H, deuterium, CN, halogen, C 1-6 Alkyl and C 3-8 Cycloalkyl.

12. The pharmaceutical composition according to any one of claims 1 to 11, which is a compound represented by formula I-3 or a pharmaceutically acceptable salt thereof, where R 9 is OH or NH2, wherein the NH2 is optionally replaced by 1 or 2 R a’ or R a” Replacement, R a’ and R a” Each independently is C 1-6 alkyl; R 10 Each independently selected from OH, halogen, CN, NH2, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl and C 1-6 haloalkoxy; m is 0, 1, or 2; R 7 Selected from H, F, Cl and C 1-6 alkyl; R 3 selected from H, F and Cl; R 4 Selected from H, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 Cycloalkyl and 3-12 membered heterocycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 The cycloalkyl group and the 3-12 membered heterocycloalkyl group are each independently optionally substituted with one or more R b or deuterium substitution; R b is selected from H, OH, CN and halogen; R 5 and R 6 Each independently selected from H, deuterium, CN, halogen, C 1-6 Alkyl and C 3-8 Cycloalkyl; Z is O or CHR 8 , R 8 is selected from the group consisting of a hydrogen atom, a deuterium atom and a halogen.

13. The pharmaceutical composition according to claim 12, wherein Z is O.

14. The pharmaceutical composition according to claim 12, wherein Z is CHR 8 , R 8 is selected from the group consisting of a hydrogen atom, a deuterium atom and a halogen.

15. The pharmaceutical composition according to claim 13 or 14, comprising a compound of formula I or a pharmaceutically acceptable salt thereof, where R 9 is OH or NH2, wherein the NH2 is optionally replaced by 1 or 2 R a’ or R a” Replacement, R a’ and R a” Each independently is C 1-6 alkyl; m is 0; R 7 Selected from H, F, Cl and C 1-6 alkyl; R 3 selected from H, F and Cl; R 4 Selected from H, C 1-6 Alkyl and C 1-6 Alkoxy, the C 1-6 Alkyl and C 1-6 Each alkoxy group is independently optionally substituted with one or more R b or deuterium substitution, R b is selected from H, OH, CN and halogen; R 5 and R 6 Each independently selected from H, deuterium, CN, halogen and C 1-6 alkyl; Preferably, where R 9 for OH; m is 0; R 7 is F or Cl; R 3 is F or Cl; R 4 H or C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with one or more R b or deuterium substitution, R b is selected from OH, CN and halogen; R 5 and R 6 Each is independently H or C 1-6 alkyl; More preferably, where R 9 for OH; m is 0; R 7 is F or Cl; R 3 is F or Cl; R 4 H or C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with one or more R b or deuterium substitution, R b for OH; R 5 and R 6 each independently selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl; Even better, where R 9 for OH; m is 0; R 7 is F or Cl; R 3 is F or Cl; R 4 is selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl, wherein the methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl are each independently optionally replaced by one or more R b Replacement; R b for OH; R 5 and R 6 each independently selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl; Particularly preferred, where R 9 for OH; m is 0; R 7 is F or Cl; R 3 is F or Cl; R 4 is selected from H, methyl, ethyl, n-propyl and isopropyl; R 5 and R 6 Each is independently selected from H, methyl, ethyl, n-propyl and isopropyl.

16. The pharmaceutical composition according to any one of claims 1 to 15, wherein the compound represented by formula I or a pharmaceutically acceptable salt thereof is selected from 17. A method for preparing a pharmaceutical composition according to any one of claims 1 to 16, comprising the steps of reacting a compound of formula IB with a compound of formula IC to form a compound of formula I. in, LG 1 is a leaving group selected from halogen, sulfonate, boronic acid and borate; X, Y, Z, L, R 1 、R 2 、R 3 、R 4 、R 5 and R 6 As defined in claim 1; Preferably, the reaction is carried out in the presence of a catalyst, which is metal palladium or metal nickel; Most preferably, the catalyst is selected from palladium / carbon, Raney nickel, tetrakistriphenylphosphine palladium, palladium dichloride, palladium acetate, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, 1,1'-[1,1'-bis(di-tert-butylphosphino)ferrocene]palladium dichloride, tris(dibenzylideneacetone)dipalladium and 2-dicyclohexylphosphine-2',6'-dimethoxybiphenyl, preferably [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride and 2-dicyclohexylphosphine-2',6'-dimethoxybiphenyl.

18. Use of the pharmaceutical composition according to any one of claims 1 to 16 in the preparation of a medicament for preventing and / or treating diseases associated with cyclin-dependent kinases; preferably, the cyclin-dependent kinase is CDK4.

19. The use according to claim 18, wherein the disease associated with cyclin-dependent kinase is selected from cell proliferation diseases, cancer and immune diseases.

20. Use of the pharmaceutical composition according to any one of claims 1 to 16 in the preparation of a medicament for preventing and / or treating cancer, wherein the cancer is selected from breast cancer, ovarian cancer, bladder cancer, uterine cancer, prostate cancer, lung cancer, esophageal cancer, head and neck cancer, intestinal cancer, kidney cancer, liver cancer, pancreatic cancer, gastric cancer and thyroid cancer; preferably, the lung cancer is selected from NSCLC, SCLC, squamous cell carcinoma or adenocarcinoma; the kidney cancer is RCC, and the liver cancer is HCC.

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