Fused tricyclic cyclin-dependent kinase inhibitors, preparation methods thereof and pharmaceutical uses

Abstract

Provided are fused tricyclic cyclin-dependent kinase inhibitors, their preparation methods and pharmaceutical uses. In particular, the fused tricyclic cyclin-dependent kinase inhibitor has a structure as shown in Formula I, wherein the definitions of each substituent are as described in the specification. The fused tricyclic cyclin-dependent kinase inhibitor is used for preventing and / or treating cyclin-dependent kinase-related diseases, especially cancers.

Description

Technical Field

[0001] The present disclosure belongs to the field of medicine and relates to a fused tricyclic cyclin-dependent kinase inhibitor, a preparation method thereof, a composition and a medical use thereof. Background Art

[0002] Cyclin-dependent kinases (CDKs) are important cellular enzymes that play important roles in regulating eukaryotic cell division and proliferation. The catalytic units of cyclin-dependent kinases are activated by regulatory subunits called cyclins. At least 16 mammalian cyclins 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 possibly other heterodynes are important regulators of 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).

[0003] In recent years, the greatest progress in the field of breast cancer treatment has undoubtedly been 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 for 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 for 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 significant clinical efficacy in estrogen receptor ER-positive metastatic breast cancer, like other kinases, their effects may be limited by the development of primary or acquired resistance over time.

[0004] Treatment with CDK4 / 6 inhibitors has been shown clinically to cause adverse reactions such as gastrointestinal and / or hematological toxicities, and acquired resistance may develop over time. Emerging data suggest that cyclin D3-CDK6 may be associated with the observed hematological 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. Thus, CDK4-selective inhibitors may offer improved safety or enhanced overall efficacy due to potentially higher and / or continuous doses compared to dual CDK4 / 6 inhibitors, and the development of highly CDK4-selective molecules has practical clinical applications. WO2019207463A discloses a class of cyclin-dependent kinase inhibitors. SUMMARY OF THE INVENTION

[0005] The present disclosure provides a compound of formula I or a pharmaceutically acceptable salt thereof,

[0006]

[0007] wherein R 1 is selected from H, C 1-6 alkyl, C 1-6 haloalkyl, and C 3-8 cycloalkyl, and the C 1-6 alkyl, C 1-6 haloalkyl, and C 3-8 cycloalkyl are each independently optionally substituted with one or more R a substituents;

[0008] R 2 is of the structure of formula II: R 9Selected from H, OH, and NH 2 , said NH 2 optionally substituted by 1 or 2 R a’ or R a ”;

[0009] R 10 each independently selected from OH, halogen, CN, NH 2 , C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-8 cycloalkyl, and 3 - 12 membered heterocycloalkyl, said C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 3-8 cycloalkyl, and 3 - 12 membered heterocycloalkyl each independently optionally substituted by one or more R b ;

[0010] Q is NR 11 or O;

[0011] Or Q is CR 12 R 13 , where R 12 , R 13 and the carbon atom to which it is attached form a 3 - 12 membered heterocycloalkyl containing N or O in NR 11 as a ring atom, said heterocycloalkyl optionally substituted by one or more R 10 ;

[0012] R 11 is selected from H, C 1-6 alkyl, C 1-6 haloalkyl, SO 2 R c , SO 2 NR d R e , COR f , COOR f , and CONR g R h , said C 1-6 alkyl and C 1-6 haloalkyl each independently optionally substituted by a group selected from R a , R b , SO 2 R c , SO 2 NR d R e , COR f , COOR f , and CONRg R h is substituted by one or more substituents of

[0013] m is 0, 1 or 2;

[0014] n is 0, 1, 2, 3 or 4;

[0015] p is 1, 2 or 3;

[0016] X is N or CH;

[0017] Y is N or CR 7 , R 7 is selected from H, F, Cl, CN, C 1-6 alkyl and C 1-6 alkoxy, and the C 1-6 alkyl and C 1-6 alkoxy are each independently optionally substituted by one or more R a substituents;

[0018] R 3 is selected from H, F, Cl, CN, CH 2 F, CHF 2 and CF 3 ;

[0019] R 4 is selected from H, C 1-6 alkyl, C 1-6 alkoxy, C3-8 cycloalkyl and 3-12 membered heterocycloalkyl, and the C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl and 3-12 membered heterocycloalkyl are each independently optionally substituted by one or more R b or deuterium;

[0020] Z is O or CHR 8 , R 8 is selected from a hydrogen atom, a deuterium atom and a halogen;

[0021] L is -(CH 2 ) q -, and the -(CH 2 )- is optionally substituted by one or more substituents selected from deuterium, CN, halogen, C 1-6 alkyl, C 3-8 cycloalkyl, C 1-6 alkoxy and 3-12 membered heterocycloalkyl, and the C 1-6 alkyl, C 3-8 cycloalkyl, C 1-6 alkoxy and 3-12 membered heterocycloalkyl are each independently optionally substituted by one or more R b or deuterium;

[0022] q is 1, 2, 3 or 4;

[0023] R 5 and R 6 are each independently selected from H, deuterium, CN, halogen, C 1-6 alkyl, C 3-8 cycloalkyl, C 1-6 alkoxy and 3- to 12-membered heterocycloalkyl, wherein the C 1-6 alkyl, C 3-8 cycloalkyl, C 1-6 alkoxy and 3- to 12-membered heterocycloalkyl are each independently optionally substituted by one or more R b or deuterium;

[0024] R a and R b are each independently selected from H, OH, CN, halogen (fluorine, chlorine, bromine, iodine), C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-8 cycloalkyl, 3- to 12-membered heterocycloalkyl and NR a’ R a” substituted, wherein the C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 3-8 cycloalkyl and 3- to 12-membered heterocycloalkyl are each independently optionally substituted by one or more substituents selected from NH 2 , NHCH 3 , N(CH 3 ) 2 , halogen, OH, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 3-8 cycloalkyl and 3- to 12-membered heterocycloalkyl;

[0025] R a’ and R a” are each independently selected from H, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl and 3- to 12-membered heterocycloalkyl, wherein the C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl and 3- to 12-membered heterocycloalkyl are each independently optionally substituted by one or more substituents selected from NH 2 , NHCH 3 , N(CH 3 ) 2 , halogen, OH, C1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-8 substituted by one or more substituents selected from cycloalkyl and 3- to 12-membered heterocycloalkyl;

[0026] or R a’ , R a” together with the N atom to which they are attached form a 3- to 12-membered heterocycloalkyl, said 3- to 12-membered heterocycloalkyl being optionally substituted by one or more substituents selected from halogen, OH, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-8 substituted by one or more substituents selected from cycloalkyl and 3- to 12-membered heterocycloalkyl;

[0027] R c , R d and R e are each independently selected from H, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl and C 1-6 haloalkyl;

[0028] R f is selected from H, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl and 3- to 12-membered heterocycloalkyl, said C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl and 3- to 12-membered heterocycloalkyl each being independently optionally substituted by one or more substituents selected from NH 2 , NHCH 3 , N(CH 3 ) 2 , halogen, OH, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 3-8 substituted by one or more substituents selected from cycloalkyl and 3- to 12-membered heterocycloalkyl;

[0029] R g and R h are each independently selected from H, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl and C 3-8 cycloalkyl, said C 1-6 alkoxy, C 1-6 haloalkyl and C3-8 Each cycloalkyl is independently optionally substituted with one or more R a or R b .

[0030] In an alternative embodiment, a compound of formula I provided by the present disclosure or a pharmaceutically acceptable salt thereof, wherein R 1 is H or C 1-6 alkyl.

[0031] In an alternative embodiment, a compound of formula I provided by the present disclosure or a pharmaceutically acceptable salt thereof, wherein R 1 is H.

[0032] In an alternative embodiment, a compound of formula I provided by the present disclosure or a pharmaceutically acceptable salt thereof, wherein R 2 is selected from

[0033]

[0034] wherein R 9 , R 10 , m and Q are as defined in the compound of formula I.

[0035] In an alternative embodiment, a compound of formula I provided by the present disclosure or a pharmaceutically acceptable salt thereof, wherein R 2 is wherein R 9 , R 10 , m and Q are as defined in the compound of formula I.

[0036] In an alternative embodiment, a compound of formula I provided by the present disclosure or a pharmaceutically acceptable salt thereof, wherein R 2 is wherein R 9 , R 10 , R 11 and m are as defined in the compound of formula I.

[0037] In an alternative embodiment, a compound of formula I provided by the present disclosure or a pharmaceutically acceptable salt thereof, wherein R 9 is OH or NH 2 , and the NH 2 is optionally substituted with 1 or 2 R a’ or R a” , and R a’ and R a” are each independently C 1-6 alkyl.

[0038] In an alternative embodiment, a compound of formula I provided by the present disclosure or a pharmaceutically acceptable salt thereof, wherein R 9is OH.

[0039] In an alternative embodiment, the compound of formula I provided by the present disclosure or a pharmaceutically acceptable salt thereof, wherein R 9 is NH 2 .

[0040] In an alternative embodiment, the compound of formula I provided by the present disclosure or a pharmaceutically acceptable salt thereof, wherein X is N.

[0041] In an alternative embodiment, the compound of formula I provided by the present disclosure or a pharmaceutically acceptable salt thereof, wherein Y is CR 7 ; R 7 is selected from H, F, Cl, and C 1-6 alkyl.

[0042] In an alternative embodiment, the compound of formula I provided by the present disclosure or a pharmaceutically acceptable salt thereof, wherein R 3 is selected from H, F, and Cl.

[0043] In an alternative embodiment, the compound of formula I provided by the present disclosure or a pharmaceutically acceptable salt thereof, wherein R 4 is selected from H, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, and 3- to 12-membered heterocycloalkyl; the C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, and 3- to 12-membered heterocycloalkyl are each independently optionally substituted with one or more R b or deuterium, and R b is selected from H, OH, CN, halogen (fluorine, chlorine, bromine, iodine), C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, and C 3-8 cycloalkyl.

[0044] In an alternative embodiment, the compound of formula I provided by the present disclosure or a pharmaceutically acceptable salt thereof, wherein R 4 is C 1-6 alkyl, and the C 1-6 alkyl is optionally substituted with one or more R b or deuterium, and R b is selected from OH, CN, halogen (fluorine, chlorine, bromine, iodine), C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, and C 3-8Naphthenyl.

[0045] In an alternative embodiment, the compound of formula I provided by the present disclosure or a pharmaceutically acceptable salt thereof, wherein R 4 is C 1-6 alkyl, and the C 1-6 alkyl is optionally substituted with one or more R b or deuterium, and R b is selected from OH, CN, and halogen (fluorine, chlorine, bromine, iodine).

[0046] In an alternative embodiment, the compound of formula I provided by the present disclosure or a pharmaceutically acceptable salt thereof, wherein L is -(CH 2 ) q -, q is selected from 1 or 2; the -(CH 2 )- is optionally substituted with one or more substituents selected from deuterium, CN, halogen, C 1-6 alkyl, C 3-8 naphthenyl, C 1-6 alkoxy, and 3- to 12-membered heterocycloalkyl.

[0047] In an alternative embodiment, the compound of formula I provided by the present disclosure or a pharmaceutically acceptable salt thereof, L is -(CH 2 ) q -, q is selected from 1; the -(CH 2 )- is optionally substituted with one or more substituents selected from deuterium, CN, halogen, C 1-6 alkyl, C 3-8 naphthenyl, C 1-6 alkoxy, and 3- to 12-membered heterocycloalkyl.

[0048] In an alternative embodiment, the compound of formula I provided by the present disclosure or a pharmaceutically acceptable salt thereof, wherein L is -(CH 2 ) qa -, q is selected from 1; the -(CH 2 )- is optionally substituted with one or more substituents of deuterium.

[0049] In an alternative embodiment, the compound of formula I provided by the present disclosure or a pharmaceutically acceptable salt thereof, wherein R 5 and R 6 are each independently selected from H, deuterium, CN, halogen, C 1-6 alkyl, and C 3-8 naphthenyl.

[0050] In an alternative embodiment, the compound of formula I provided by the present disclosure or a pharmaceutically acceptable salt thereof is the compound of formula I-2 or a pharmaceutically acceptable salt thereof,

[0051]

[0052] wherein R 9 is OH or NH 2 and the NH 2 is optionally substituted by 1 or 2 R a’ or R a” and R a’ and R a” are each independently C 1-6 alkyl;

[0053] R 10 are each independently selected from H, OH, halogen, CN, NH 2 , C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl and C 1-6 haloalkoxy;

[0054] m is 0, 1 or 2;

[0055] R 11 is selected from SO 2 R c , SO 2 NR d R e , COR f , COOR f and CONR g R h ;

[0056] R c , R d and R e are each independently selected from H, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl and C 1-6 haloalkyl;

[0057] R f is selected from H, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl and 3- to 12-membered heterocycloalkyl;

[0058] R g , R h are each independently selected from H, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl and C 3-8 cycloalkyl;

[0059] R 7 is selected from H, F, Cl and C 1-6 alkyl;

[0060] R 3 selected from H, F, and Cl;

[0061] R 4 selected from H, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, and 3- to 12-membered heterocycloalkyl; the C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, and 3- to 12-membered heterocycloalkyl are each independently optionally substituted with one or more R b or deuterium, and R b is selected from H, OH, CN, and halogen (fluorine, chlorine, bromine, iodine);

[0062] R 5 and R 6 are each independently selected from H, deuterium, CN, halogen, C 1-6 alkyl, and C 3-8 cycloalkyl;

[0063] Z is O or CHR 8 , and R 8 is selected from a hydrogen atom, a deuterium atom, and a halogen.

[0064] In an alternative embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is the compound of formula I-2 or a pharmaceutically acceptable salt thereof 2, wherein Z is O.

[0065] In an alternative embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is the compound of formula I-2 or a pharmaceutically acceptable salt thereof, wherein Z is CHR 8 , and R 8 is selected from a hydrogen atom, a deuterium atom, and a halogen.

[0066] In an alternative embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is the compound of formula I-2 or a pharmaceutically acceptable salt thereof,

[0067] wherein R 9 is OH or NH 2 , and the NH 2 is optionally substituted with 1 or 2 R a’ or R a” , and R a’ and R a” are each independently C 1-6 alkyl;

[0068] m is 0;

[0069] R11 is SO 2 R c ;

[0070] R c is selected from H, C 1-6 alkyl and C 1-6 haloalkyl;

[0071] R 7 is selected from H, F, Cl and C 1-6 alkyl;

[0072] R 3 is selected from H, F and Cl;

[0073] R 4 is selected from H, C 1-6 alkyl and C 1-6 alkoxy; the C 1-6 alkyl and C 1-6 alkoxy are each independently optionally substituted by one or more R b or deuterium; R b is selected from H, OH, CN and halogen (fluorine, chlorine, bromine, iodine);

[0074] R 5 and R 6 are each independently selected from H, deuterium, CN, halogen and C 1-6 alkyl.

[0075] In an alternative embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is the compound of formula I-2 or a pharmaceutically acceptable salt thereof,

[0076] wherein R 9 is OH;

[0077] m is 0;

[0078] R 11 is SO 2 R c ;

[0079] R c is C 1-6 alkyl or C 1-6 haloalkyl;

[0080] R 7 is F or Cl;

[0081] R 3 is F or Cl;

[0082] R 4 is H or C 1-6 alkyl, the C 1-6 alkyl is optionally substituted by one or more Rb or deuterium substitution, R b is selected from H, OH, CN, and halogen (fluorine, chlorine, bromine, iodine);

[0083] R 5 and R 6 are each independently H or C 1-6 alkyl.

[0084] In an alternative embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is the compound of formula I-2 or a pharmaceutically acceptable salt thereof,

[0085] wherein R 9 is OH;

[0086] m is 0;

[0087] R 11 is SO 2 R c ;

[0088] R c is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and tert-butyl;

[0089] R 7 is F or Cl;

[0090] R 3 is F or Cl;

[0091] R 4 is H or C 1-6 alkyl, and the C 1-6 alkyl is optionally substituted with one or more R b or deuterium, and R b is H or OH;

[0092] R 5 and R 6 are each independently selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and tert-butyl.

[0093] In an alternative embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is the compound of formula I-2 or a pharmaceutically acceptable salt thereof,

[0094] wherein R 9 is OH;

[0095] m is 0;

[0096] R 11 is SO 2 R c ;

[0097] R cSelected from methyl, ethyl, and n-propyl;

[0098] R 7 is F or Cl;

[0099] R 3 is F or Cl;

[0100] R 4 is selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and tert-butyl, and the methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and tert-butyl are each independently optionally substituted with one or more R b substituents, and R b is H or OH;

[0101] R 5 and R 6 are each independently selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and tert-butyl.

[0102] In an optional embodiment, the compound of formula I provided by the present disclosure or a pharmaceutically acceptable salt thereof is the compound of formula I-2 or a pharmaceutically acceptable salt thereof,

[0103] wherein R 9 is OH;

[0104] m is 0;

[0105] R 11 is SO 2 R c ;

[0106] R c is methyl;

[0107] R 7 is F or Cl;

[0108] R 3 is F or Cl;

[0109] R 4 is selected from H, methyl, ethyl, n-propyl, and isopropyl;

[0110] R 5 and R 6 are each independently selected from H, methyl, ethyl, n-propyl, and isopropyl.

[0111] In an optional embodiment, the compound of formula I provided by the present disclosure or a pharmaceutically acceptable salt thereof is the compound of formula I-3 or a pharmaceutically acceptable salt thereof,

[0112]

[0113] wherein R 9is OH or NH 2 , said NH 2 is optionally substituted by 1 or 2 Rs a’ or R a” , R a’ and R a” are each independently C 1-6 alkyl;

[0114] R 10 are each independently selected from OH, halogen, CN, NH 2 , C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl and C 1-6 haloalkoxy;

[0115] m is 0, 1 or 2;

[0116] R 7 is selected from H, F, Cl and C 1-6 alkyl;

[0117] R 3 is selected from H, F and Cl;

[0118] R 4 is selected from H, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl and 3-12 membered heterocycloalkyl, said C 1-6 alkyl, C 1-6 alkoxy, C3-8 cycloalkyl and 3-12 membered heterocycloalkyl are each independently optionally substituted by one or more Rs b or deuterium; R b is selected from H, OH, CN and halogen (fluorine, chlorine, bromine, iodine);

[0119] R 5 and R 6 are each independently selected from H, deuterium, CN, halogen, C 1-6 alkyl and C 3-8 cycloalkyl;

[0120] Z is O or CHR 8 , R 8 is selected from a hydrogen atom, a deuterium atom and a halogen.

[0121] In an optional embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is the compound of formula I-3 or a pharmaceutically acceptable salt thereof, wherein Z is O.

[0122] In an alternative embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is the 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.

[0123] In an alternative embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is the compound of formula I-3 or a pharmaceutically acceptable salt thereof,

[0124] wherein R 9 is OH or NH 2 , and the NH 2 is optionally substituted by 1 or 2 R a’ or R a” , and R a’ and R a” are each independently C 1-6 alkyl;

[0125] m is 0;

[0126] R 7 is selected from H, F, Cl and C 1-6 alkyl;

[0127] R 3 is selected from H, F and Cl;

[0128] R 4 is selected from H, C 1-6 alkyl and C 1-6 alkoxy; the C 1-6 alkyl and C 1-6 alkoxy are each independently optionally substituted by one or more R b or deuterium, and R b is selected from H, OH, CN and halogen (fluorine, chlorine, bromine, iodine);

[0129] R 5 and R 6 are each independently selected from H, deuterium, CN, halogen and C 1-6 alkyl.

[0130] In an alternative embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is the compound of formula I-3 or a pharmaceutically acceptable salt thereof,

[0131] wherein R 9 is OH;

[0132] m is 0;

[0133] R 7 is F or Cl;

[0134] R 3 is F or Cl;

[0135] R 4 is H or C 1-6 alkyl, and the C 1-6 alkyl is optionally substituted by one or more R b or deuterium, and R b is selected from H, OH, CN, and halogen (fluorine, chlorine, bromine, iodine);

[0136] R 5 and R 6 are each independently H or C 1-6 alkyl.

[0137] In an alternative embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is the compound of formula I-3 or a pharmaceutically acceptable salt thereof,

[0138] wherein R 9 is OH;

[0139] m is 0;

[0140] R 7 is F or Cl;

[0141] R 3 is F or Cl;

[0142] R 4 is H or C 1-6 alkyl, and the C 1-6 alkyl is optionally substituted by one or more R b or deuterium, and R b is H or OH;

[0143] R 5 and R 6 are each independently selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and tert-butyl.

[0144] In an alternative embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is the compound of formula I-3 or a pharmaceutically acceptable salt thereof,

[0145] wherein R 9 is OH;

[0146] m is 0;

[0147] R 7 is F or Cl;

[0148] R 3 is F or Cl;

[0149] R 4 Selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl, and the methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl are each independently optionally substituted by one or more R b substituted; R b is H or OH;

[0150] R 5 and R 6 are each independently selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl.

[0151] In an optional embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is the compound of formula I-3 or a pharmaceutically acceptable salt thereof,

[0152] wherein R 9 is OH;

[0153] m is 0;

[0154] R 7 is F or Cl;

[0155] R 3 is F or Cl;

[0156] R 4 is selected from H, methyl, ethyl, n-propyl and isopropyl;

[0157] R 5 and R 6 are each independently selected from H, methyl, ethyl, n-propyl and isopropyl.

[0158] In an optional embodiment, the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present disclosure is selected from

[0159]

[0160]

[0161]

[0162] On the other hand, the present disclosure provides an isotope-substituted compound of the compound of formula I, I-2 or I-3 or a pharmaceutically acceptable salt thereof, and in an optional embodiment, the isotope-substituted compound is a deuterium atom-substituted compound.

[0163] In an optional embodiment, for the compound of formula I or a pharmaceutically acceptable salt thereof, the abundance of the deuterium atom is greater than 20%.

[0164] In an alternative embodiment, for the compound of formula I or a pharmaceutically acceptable salt thereof, the abundance of the deuterium atom is greater than 50%.

[0165] In an alternative embodiment, for the compound of formula I or a pharmaceutically acceptable salt thereof, the abundance of the deuterium atom is greater than 90%.

[0166] In an alternative embodiment, for the compound of formula I or a pharmaceutically acceptable salt thereof, the abundance of the deuterium atom is greater than 95%.

[0167] The present disclosure also provides a method for preparing the compound of formula I, which includes the step of reacting the compound of formula I-B with the compound of formula I-C to form the compound of formula I,

[0168]

[0169] wherein LG 1 is a leaving group, and the leaving group is preferably halogen, sulfonate, boric acid, and borate;

[0170] X, Y, Z, L, R 1 、R 2 、R 3 、R 4 、R 5 and R 6 are as defined in the compound of formula I.

[0171] In some embodiments, the reaction is carried out in the presence of a catalyst, and the catalyst is palladium metal or nickel metal.

[0172] In some embodiments, the catalyst is selected from palladium / carbon, Raney nickel, tetrakis(triphenylphosphine)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-bis(cyclohexylphosphino)-2',6'-dimethoxybiphenyl, preferably [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium and 2-bis(cyclohexylphosphino)-2',6'-dimethoxybiphenyl.

[0173] On the other hand, the present disclosure provides a compound of formula I-B or a pharmaceutically acceptable salt thereof,

[0174]

[0175] wherein LG 1 is a leaving group, and the leaving group is selected from halogen, sulfonate, boric acid, and borate; X, Y, Z, L, R 3 、R 4 、R 5 、R 6As defined in the compounds of Formula I.

[0176] The present disclosure also provides a pharmaceutical composition comprising at least one therapeutically effective amount of the compounds of Formula I, I-2, or I-3 as described above, or a pharmaceutically acceptable salt or the aforementioned isotope-substituted compound thereof, and a pharmaceutically acceptable excipient.

[0177] In some embodiments, the unit dose of the pharmaceutical composition is 0.001 mg - 1000 mg.

[0178] In certain embodiments, based on the total weight of the composition, the pharmaceutical composition contains 0.01 - 99.99% of the compounds of Formula I, I-2, or I-3 as described above, or a pharmaceutically acceptable salt or the aforementioned isotope-substituted compound thereof.

[0179] In certain embodiments, the pharmaceutical composition contains 0.1 - 99.9% of the compounds of Formula I, I-2, or I-3 as described above, or a pharmaceutically acceptable salt or the aforementioned isotope-substituted compound thereof.

[0180] In certain embodiments, the pharmaceutical composition contains 0.5% - 99.5% of the compounds of Formula I, I-2, or I-3 as described above, or a pharmaceutically acceptable salt or the aforementioned isotope-substituted compound thereof.

[0181] In certain embodiments, the pharmaceutical composition contains 1% - 99% of the compounds of Formula I, I-2, or I-3 as described above, or a pharmaceutically acceptable salt or the aforementioned isotope-substituted compound thereof.

[0182] In certain embodiments, the pharmaceutical composition contains 2% - 98% of the compounds of Formula I, I-2, or I-3 as described above, or a pharmaceutically acceptable salt or the aforementioned isotope-substituted compound thereof.

[0183] In certain embodiments, based on the total weight of the composition, the pharmaceutical composition contains 0.01% - 99.99% of a pharmaceutically acceptable excipient.

[0184] In certain embodiments, the pharmaceutical composition contains 0.1% - 99.9% of a pharmaceutically acceptable excipient.

[0185] In certain embodiments, the pharmaceutical composition contains 0.5% - 99.5% of a pharmaceutically acceptable excipient.

[0186] In certain embodiments, the pharmaceutical composition contains 1% - 99% of a pharmaceutically acceptable excipient.

[0187] In certain embodiments, the pharmaceutical composition contains 2% - 98% of a pharmaceutically acceptable excipient.

[0188] The present disclosure also provides a method for preventing and / or treating cyclin-dependent kinase-related diseases, which comprises administering to a patient in need thereof a therapeutically effective amount of the compound represented by the foregoing formula I, I-2, I-3, or a pharmaceutically acceptable salt thereof, or the foregoing isotope-substituted compound, or the foregoing pharmaceutical composition.

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

[0190] The present disclosure also provides the use of the compound represented by the foregoing formula I, I-2, I-3, or a pharmaceutically acceptable salt thereof, or the foregoing isotope-substituted compound, or the foregoing pharmaceutical composition in the preparation of a medicament for preventing and / or treating cyclin-dependent kinase-related diseases.

[0191] The present disclosure also provides the use of the compound represented by the foregoing formula I, I-2, I-3, or a pharmaceutically acceptable salt thereof, or the foregoing isotope-substituted compound, or the foregoing pharmaceutical composition in the preparation of a medicament for preventing and / or treating cancer.

[0192] In an alternative embodiment, the cyclin-dependent kinase-related diseases are selected from cell proliferative diseases, cancer, and immune diseases.

[0193] 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.

[0194] In an alternative embodiment, the cyclin-dependent kinase in the present disclosure is CDK4.

[0195] On the other hand, the present disclosure provides a use of the compound represented by the foregoing formula I, I-2, I-3, or a pharmaceutically acceptable salt thereof

[0196] or the foregoing isotope-substituted compound as a medicament.

[0197] The compound represented by the foregoing formula I, I-2, I-3, or a pharmaceutically acceptable salt thereof, or the foregoing isotope-substituted compound, or the foregoing pharmaceutical composition provided by the present disclosure reduces gastrointestinal and / or hematological toxicity.

[0198] On the other hand, the pharmaceutically acceptable salts of the compounds 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 their racemic mixtures and other mixtures, such as enantiomer- or diastereomer- enriched mixtures, all of which mixtures are within the scope of the present disclosure. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and their mixtures are included within the scope of the present disclosure. Compounds of the present disclosure containing asymmetric carbon atoms may be isolated in optically pure form or as a racemate. The optically pure form may be resolved from the racemic mixture or synthesized by using chiral starting materials or chiral reagents.

[0200] The optically active (R)- and (S)- isomers and D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If an 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 a suitable optically active acid or base, and then the diastereomers are resolved by conventional methods known in the art, and then the pure enantiomer is recovered. In addition, the separation of enantiomers and diastereomers is usually accomplished by using chromatography employing a chiral stationary phase and optionally in combination with chemical derivatization (such as formation of a carbamate from an amine).

[0201] In the chemical structure of the compounds of the present disclosure, the bond represents an unspecified configuration, that is, if chiral isomers are present in the chemical structure, the bond can be or can simultaneously contain both configurations. The bond represents an unspecified configuration, including cis (E) or trans (Z) configurations.

[0202] The compounds and intermediates of the present disclosure may also exist in different tautomeric forms, and all such forms are included within 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 known as prototropic tautomers) include interconversions via proton migration, such as keto - enol and imine - enamine, lactam - lactim isomerization. An example of a lactam - lactim equilibrium is between A and B shown below.

[0203]

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

[0205] The present disclosure also includes isotopically labeled compounds of the present disclosure that are the same as those described herein, but in which one or more atoms are replaced with atoms having an atomic weight or mass number different from the atomic weight or mass number normally 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 etc.

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

[0207] Term Explanation:

[0208] "Pharmaceutically acceptable excipients" include, but are not limited to, any adjuvant, carrier, excipient, glidant, sweetening agent, diluent, preservative, dye / colorant, flavoring agent, surfactant, wetting agent, dispersing agent, suspending agent, stabilizing agent, isotonic agent, solvent or emulsifying agent that has been approved by the US Food and Drug Administration for use in humans or domestic animals.

[0209] "Alkyl" refers to saturated aliphatic hydrocarbon groups, including straight-chain and branched-chain groups having from 1 to 20 carbon atoms. Alkyl groups having from 1 to 12 carbon atoms are preferred, and alkyl groups having from 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 isomers thereof, etc. The alkyl group can be substituted or unsubstituted, and when substituted, the substituent can be substituted at any available attachment point, 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- to 12-membered heterocyclic groups, etc.

[0210] "Alkenyl" includes branched and straight-chain alkenes having from 2 to 12 carbon atoms or alkenes containing aliphatic hydrocarbon groups. For example, "C 2-6 alkenyl" represents 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. The alkenyl group can be substituted or unsubstituted, and when substituted, the substituent can be substituted at any available attachment point, 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- to 12-membered heterocyclic groups, etc.

[0211] "Alkynyl" includes branched and straight-chain alkynyl groups having from 2 to 12 carbon atoms or alkenes containing aliphatic hydrocarbon groups, or if a specific number of carbon atoms is specified, it means that particular number. For example, ethynyl, propynyl (e.g., 1-propynyl, 2-propynyl), 3-butynyl, pentynyl, hexynyl, and 1-methylpent-2-ynyl. The alkynyl group can be substituted or unsubstituted, and when substituted, the substituent can be substituted at any available attachment point, preferably one or more of the following groups, independently selected from halogen, hydroxy, oxo, nitro, cyano, C 1-6 alkyl, C1-6 Alkoxy, C 3-7 Cycloalkyl, 3- to 12-membered heterocyclic group, etc.

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

[0213] The cycloalkyl ring may be fused to an aryl, heteroaryl, or heterocycloalkyl ring, where the ring attached to the parent structure is cycloalkyl, non-limiting examples including indanyl, tetrahydronaphthyl, benzocycloheptyl, etc. Cycloalkyl 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 3-7 Cycloalkyl, 3- to 12-membered heterocyclic group, etc.

[0214] The term "heterocyclic group", also expressed as heterocycloalkyl, refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent containing 3 to 20 ring atoms, one or more of which are heteroatoms selected from nitrogen, oxygen, or S(O) m (where m is an integer from 0 to 2), but not including ring moieties of -O-O-, -O-S-, or -S-S-, and the remaining ring atoms are carbon. Preferably, it contains 3 to 12 ring atoms, where 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 spiro, fused, and bridged heterocyclic groups. Non-limiting examples of "heterocyclic group" include:

[0215]

[0216] And so on.

[0217] The heterocyclic group ring may be fused to an aryl, heteroaryl, or cycloalkyl ring, where the ring attached to the parent structure is heterocyclic group, non-limiting examples of which include:

[0218] And so on.

[0219] The heterocyclic group may be optionally substituted or unsubstituted. 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- to 12-membered heterocyclic group, etc.

[0220] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., rings sharing adjacent carbon atom pairs) group having a conjugated π electron system, preferably 6 to 12 members, such as phenyl and naphthyl. The aryl ring may be fused to a heteroaryl, heterocyclic or cycloalkyl ring, where the ring connected to the parent structure is the aryl ring. Non-limiting examples thereof include:

[0221]

[0222] The aryl may be substituted or unsubstituted. 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- to 12-membered heterocyclic group, etc., preferably phenyl.

[0223] The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 14 ring atoms, where the heteroatoms are selected from oxygen, sulfur and nitrogen. The heteroaryl is preferably 6 to 12 members, more preferably 5 or 6 members. For example. Non-limiting examples thereof include: imidazolyl, furyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, pyrrolyl, tetrazolyl, pyridyl, pyrimidinyl, thiadiazole, pyrazine, and so on.

[0224] The heteroaryl ring may be fused to an aryl, heterocyclic or cycloalkyl ring, where the ring connected to the parent structure is the heteroaryl ring. Non-limiting examples thereof include:

[0225]

[0226] The heteroaryl may be optionally substituted or unsubstituted. 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- to 12-membered heterocyclic group, etc.

[0227] The term "alkoxy" refers to -O-(alkyl) and -O-(unsubstituted cycloalkyl), where the definition of alkyl is as described above. Non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy. The alkoxy can be optionally substituted or unsubstituted. 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- to 12-membered heterocyclic group, etc.

[0228] The term "hydroxy" refers to the -OH group.

[0229] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0230] The term "amino" refers to -NH 2 .

[0231] The term "cyano" refers to -CN.

[0232] The term "nitro" refers to -NO 2 .

[0233] The term "oxo" refers to the ═O substituent.

[0234] "Optional" or "optionally" means that the subsequent described event or circumstance can but does not have to occur, and this description includes the occasions where the event or circumstance occurs or does not occur. For example, "a heterocyclic group optionally substituted by alkyl" means that alkyl can but does not have to be present, and this description includes the case where the heterocyclic group is substituted by alkyl and the case where the heterocyclic group is not substituted by alkyl.

[0235] "Substituted" means that one or more hydrogen atoms in the group, preferably up to 5, more preferably 1 to 3 hydrogen atoms, are independently replaced by the corresponding number of substituents. It goes without saying that the substituents are only at their possible chemical positions, and those skilled in the art can determine (by experiment or theory) the possible or impossible substitutions without excessive effort. For example, an amino or hydroxy group with a free hydrogen may be unstable when combined with a carbon atom having an unsaturated (such as olefinic) bond.

[0236] "Pharmaceutical composition" means a mixture containing one or more compounds described herein or their physiologically and pharmaceutically acceptable salts or prodrugs and other chemical components, as well as other components such as physiologically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to facilitate the administration to an organism, facilitate the absorption of the active ingredient and thus exert its biological activity. Detailed Description

[0237] The present disclosure will be further described below in conjunction with embodiments, but these embodiments do not limit the scope of the present disclosure.

[0238] In the embodiments of the present disclosure, the experimental methods without specific conditions are generally carried out according to conventional conditions or according to the conditions recommended by the raw material or commodity manufacturers. The reagents without specific sources are conventional reagents purchased from the market.

[0239] The structure of the compound is determined by nuclear magnetic resonance (NMR) or / and mass spectrometry (MS). The NMR shift (δ) is given in units of 10 -6 (ppm). The NMR measurement is performed using a Bruker AVANCE-400 nuclear magnetic resonance spectrometer, and the measurement solvent is deuterated dimethyl sulfoxide (DMSO-d 6 ), deuterated chloroform (CDCl 3 ), deuterated methanol (CD 3 OD), and the internal standard is tetramethylsilane (TMS).

[0240] The MS measurement is performed using a Shimadzu 2010 Mass Spectrometer or an Agilent 6110A MSD mass spectrometer.

[0241] The HPLC measurement is performed using a Shimadzu LC-20A systems, a Shimadzu LC-2010HT series, or an Agilent 1200 LC high-performance liquid chromatograph (Ultimate XB-C18 3.0*150mm chromatographic column or Xtimate C18 2.1*30mm chromatographic column).

[0242] The chiral HPLC analysis and determination are performed using Chiralpak IC-3 100×4.6mm I.D., 3um, Chiralpak AD-3 150×4.6mm I.D., 3um, Chiralpak AD-3 50×4.6mm I.D., 3um, Chiralpak AS-3 150×4.6mm I.D., 3um, Chiralpak AS-3 100×4.6mm I.D., 3μm, ChiralCel OD-3 150×4.6mm I.D., 3um, Chiralcel OD-3 100×4.6mm I.D., 3μm, ChiralCel OJ-H 150×4.6mm I.D., 5um, Chiralcel OJ-3 150×4.6mm I.D., 3um chromatographic columns;

[0243] For thin-layer chromatography silica gel plates, Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates are used. The specifications of the silica gel plates used in thin-layer chromatography (TLC) are 0.15 mm to 0.2 mm, and the specifications of the silica gel plates used for thin-layer chromatography separation and purification of products are 0.4 mm to 0.5 mm.

[0244] For column chromatography, silica gel with 100 - 200 mesh, 200 - 300 mesh or 300 - 400 mesh from Yantai Huanghai is generally used as the carrier.

[0245] For chiral preparation columns, DAICEL CHIRALPAK IC (250 mm * 30 mm, 10 μm) or Phenomenex - Amylose - 1 (250 mm * 30 mm, 5 μm) are used.

[0246] For the CombiFlash rapid preparation instrument, Combiflash Rf150 (TELEDYNE ISCO) is used.

[0247] The average inhibition rate of kinase and IC 50 value is measured using a NovoStar microplate reader (from BMG Labtech, Germany).

[0248] The known starting materials of the present disclosure can be adopted or synthesized according to methods known in the art, or can be purchased from companies such as ABCR GmbH&Co.KG, Acros Organics, Aldrich Chemical Company, AccelaChemBio Inc, Darui Chemicals, etc.

[0249] Unless otherwise specified in the examples, the reactions can all be carried out under an argon or nitrogen atmosphere.

[0250] An argon or nitrogen atmosphere means that the reaction flask is connected to an argon or nitrogen balloon with a volume of about 1 L.

[0251] A hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon with a volume of about 1 L.

[0252] For catalytic hydrogenation reactions, a Parr 3916EKX hydrogenator and Qinglan QL - 500 hydrogen gas generator or an HC2 - SS hydrogenator are used.

[0253] For hydrogenation reactions, usually, the system is evacuated, filled with hydrogen gas, and this operation is repeated 3 times.

[0254] For microwave reactions, a CEM Discover - S 908860 microwave reactor is used.

[0255] Unless otherwise specified in the examples, the solution refers to an aqueous solution.

[0256] Unless otherwise specified in the examples, the reaction temperature is room temperature, which is 20°C to 30°C.

[0257] The progress of the reaction in the examples was monitored by thin-layer chromatography (TLC).

[0258] For the eluent used in the reaction, the eluent system of column chromatography for purifying the compound, and the eluent system of thin-layer chromatography, the volume ratio of the solvents was adjusted according to the polarity of the compound, and a small amount of basic or acidic reagents such as triethylamine and acetic acid could also be added for adjustment.

[0259] Example 1

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

[0261] (3S,4R)-4-((5-Fluoro-4-(-8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazacyclopenta[a]acenaphthylen-6-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol isomer 2

[0262]

[0263] The first step

[0264] 6-Bromo-4-fluoro-2,3-dinitrophenol 1b

[0265] Compound 1a (3.5 g, 14.8 mmol) was dissolved in 16 mL of dichloromethane. A dichloromethane solution of nitric acid (2 mol / L, 16 mL) was added. The reaction was carried out at room temperature for 20 minutes. The reaction solution was poured into 50 mL of ice water, the organic phase was separated, the aqueous phase was extracted with dichloromethane (50 mL × 2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was collected, and the filtrate was concentrated under reduced pressure to obtain the title compound 1b (3.8 g, yield: 91%).

[0266] MS(ESI) m / z 279.0, 281.0 [M-H] -

[0267] The second step

[0268] 2-Amino-6-bromo-4-fluoro-3-nitrophenol 1c

[0269] 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 in portions. The reaction was carried out 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 × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was collected, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with petroleum ether and ethyl acetate to obtain the title compound 1c (2.0 g, yield: 59%).

[0270] MS(ESI) m / z 249.1, 251.1 [M-H] -

[0271] The third step

[0272] 8-Bromo-6-fluoro-3-methyl-5-nitro-3,4-dihydro-2H-benzo[b][1,4]oxazine 1d

[0273] 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 the reaction was carried out at room temperature for 2 hours. 20 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was collected, and the filtrate was concentrated under reduced pressure. 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 in portions. The reaction was carried out at room temperature for 2 hours. The reaction solution was poured into 20 mL of saturated sodium bicarbonate solution, and the mixture was extracted with ethyl acetate (20 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was collected, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with petroleum ether and ethyl acetate to obtain the title compound 1d (160 mg, yield: 69%).

[0274] MS(ESI) m / z 291.2, 293.2 [M+H] +

[0275] The fourth step

[0276] 8-Bromo-6-fluoro-3-methyl-3,4-dihydro-2H-benzo[b][1,4]oxazin-5-amine 1e

[0277] 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 carried out 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×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 eluting with petroleum ether and ethyl acetate to obtain the title compound 1e (105 mg, yield: 73%).

[0278] MS(ESI) m / z 261.3, 263.3 [M+H] +

[0279] The fifth step

[0280] 6-Bromo-8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazacenaphthylene 1f

[0281] 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 reaction was carried out at 120 °C for 2 hours. The reaction mixture was concentrated, 30 mL of saturated sodium bicarbonate solution was added, and extracted with ethyl acetate (30 mL×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%).

[0282] MS(ESI) m / z 241.3, 243.3 [M+H] +

[0283] The sixth step

[0284] 6-(2-Chloro-5-fluoropyrimidin-4-yl)-8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazacenaphthylene 1g

[0285] Under a nitrogen atmosphere, compound 1f (40 mg, 0.14 mmol), bis(pinacolato)diboron (57 mg, 0.22 mmol), potassium acetate (29 mg, 0.29 mmol), and dichlorobis(1,1'-bis(diphenylphosphino)ferrocene)palladium(II) (21 mg, 0.03 mmol) were successively dissolved in 2 mL of 1,4-dioxane. The reaction was carried out at 100 °C for 1 hour. The reaction solution was cooled to room temperature, and 1,3,5,7-tetramethyl-6-phenyl-2,4,8-trioxa-6-phosphaadamantane (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(0) (26 mg, 0.03 mmol), and 0.5 mL of water were added. 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. After concentration under reduced pressure, the residue was purified by C-18 reverse-phase chromatography to obtain the title compound 1g (18 mg, yield: 38%).

[0286] MS(ESI) m / z 337.2 [M+H] +

[0287] The seventh step

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

[0289] (3S,4R)-4-((5-Fluoro-4-(-8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylen-6-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol isomer 2

[0290] 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”-binaphthalene (6.2 mg, 0.01 mmol), and palladium(II) acetate (2.2 mg, 0.01 mmol) were successively dissolved in 2 mL of tetrahydrofuran. Cesium carbonate (41 mg, 0.13 mmol) was added, and the reaction was carried out at 80 °C for 1 hour.

[0291] 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 product mixture. The crude product was subjected to chiral resolution (column: DAICEL CHIRALPAK AD (250 mm * 30 mm, 10 μm), conditions: 45% EtOH (0.1% NH3· H 2 O) in CO 2 medium; flow rate: 80 mL / min) to obtain isomer 1 (3.8 mg, yield: 17%) and isomer 2 (4.4 mg, yield: 20%).

[0292] Analysis method

[0293] Column: DAICEL CHIRALPAK AD-3 (150 mm * 4.6 mm, 3 μm);

[0294] Condition: CO 2 condition, 40% EtOH (0.05% DEA);

[0295] Flow rate: 2.5 mL / min;

[0296] ABPR: 1500 psi;

[0297] Temperature: 35 °C.

[0298] The compound with a retention time of 2.903 min was defined as isomer 1; MS (ESI) m / z 418.3 [M + H] +

[0299] The compound with a retention time of 3.997 min was defined as isomer 2; MS (ESI) m / z 418.3 [M + H] +

[0300] Example 2

[0301] (3R,4R)-4-((5-chloro-4-(8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazacyclopenta[a]acenaphthylen-6-yl)pyrimidin-2-yl)amino)-1-(methanesulfonyl)piperidin-3-ol isomer 1

[0302] (3R,4R)-4-((5-chloro-4-(8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazacyclopenta[a]acenaphthylen-6-yl)pyrimidin-2-yl)amino)-1-(methanesulfonyl)piperidin-3-ol isomer 2

[0303]

[0304] The first step

[0305] 6-(2,5-dichloropyrimidin-4-yl)-8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazacyclopenta[a]acenaphthylene 2a

[0306] Under a nitrogen atmosphere, compound 1f (50 mg, 0.18 mmol), bis(pinacolato)diboron (67 mg, 0.26 mmol), potassium acetate (37 mg, 0.38 mmol), and palladium(II) dichloride [1,1'-bis(diphenylphosphino)ferrocene] (26 mg, 0.04 mmol) were successively dissolved in 2 mL of 1,4-dioxane. The reaction was carried out at 100 °C for 1 hour. After the reaction solution was cooled to room temperature, 1,3,5,7-tetramethyl-6-phenyl-2,4,8-trioxa-6-phosphaadamantane (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(0) (32 mg, 0.04 mmol), and 0.5 mL of water were 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. After concentration under reduced pressure, the residue was purified by C-18 reverse-phase chromatography to obtain the title compound 2a (30 mg, yield: 48%).

[0307] MS(ESI) m / z 353.1 [M+H] +

[0308] The second step

[0309] (3R,4R)-4-((5-chloro-4-(8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazacyclopenta[c]acenaphthylen-6-yl)pyrimidin-2-yl)amino)-1-(methanesulfonyl)piperidin-3-ol isomer 1

[0310] (3R,4R)-4-((5-chloro-4-(8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazacyclopenta[c]acenaphthylen-6-yl)pyrimidin-2-yl)amino)-1-(methanesulfonyl)piperidin-3-ol isomer 2

[0311] 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 by the method disclosed in patent application "WO 2019 / 207463 A1"), (S)-(-)-2,2”-bis(diphenylphosphino)-1,1”-binaphthalene (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. After concentration under reduced pressure, the residue was purified by C-18 reverse-phase chromatography to obtain a crude product mixture. The crude product was subjected to chiral resolution (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 obtain isomer 1 (4.4 mg, yield: 10%) and isomer 2 (3.3 mg, yield: 8%).

[0312] Analysis method

[0313] Column: DAICEL CHIRALPAK AD-3 (50 mm * 4.6 mm, 3 μm);

[0314] Mobile phase: A: CO 2 B: isopropanol (0.05% DEA), gradient: 5% to 40% of B in 2 min, maintain 40% of B for 1.2 min, then 5% of B for 0.8 min;

[0315] Flow rate: 4 mL / min;

[0316] ABPR: 1500 psi;

[0317] Temperature: 35 °C.

[0318] The compound with a retention time of 2.094 min was defined as isomer 1;

[0319] MS(ESI) m / z 511.3 [M+H] +

[0320] 11H NMR (400 MHz, DMSO-d6) δ = 8.38 (s, 1H), 7.48 (br s, 1H), 6.91 (br d, J = 11.0 Hz, 1H), 5.21 (br s, 1H), 4.86 (br d, J = 6.5 Hz, 1H), 4.52 - 4.44 (m, 1H), 4.24 (br d, J = 10.8 Hz, 1H), 3.77 (br s, 1H), 3.64 - 3.55 (m, 2H), 3.48 (br d, J = 13.6 Hz, 1H), 2.89 (s, 3H), 2.69 - 2.63 (m, 1H), 2.60 (s, 3H), 1.39 (d, J = 6.8 Hz, 3H), 1.24 (br s, 2H), 1.18 - 1.04 (m, 1H).

[0321] The compound with a retention time of 2.499 min was defined as isomer 2;

[0322] MS (ESI) m / z 511.3 [M + H] +

[0323] 1 1H NMR (400 MHz, DMSO-d6) δ = 8.38 (s, 1H), 7.47 (br s, 1H), 6.91 (br d, J = 11.5 Hz, 1H), 5.21 (br d, J = 4.0 Hz, 1H), 4.86 (br d, J = 6.5 Hz, 1H), 4.48 (dd, J = 1.8, 11.5 Hz, 1H), 4.36 (t, J = 5.0 Hz, 2H), 4.24 (br d, J = 9.5 Hz, 1H), 3.76 (br s, 1H), 3.59 (br d, J = 8.0 Hz, 2H), 2.89 (s, 3H), 2.67 (br d, J = 9.3 Hz, 1H), 2.60 (s, 3H), 1.58 - 1.46 (m, 1H), 1.39 (d, J = 6.8 Hz, 3H), 1.24 (br s, 1H).

[0324] Example 3

[0325] (3S,4R)-4-((5-chloro-4-((S)-8-fluoro-2,3-dimethyl-3,4-dihydro-5-oxa-1,2a-diazacenaphthylen-6-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-diazacenaphthylen-6-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol isomer 2

[0327]

[0328] 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”-binaphthalene (20 mg, 0.03 mmol), and palladium(II) acetate (6.7 mg, 0.03 mmol) were successively 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 product mixture. The crude product was subjected to chiral resolution [column: DAICEL CHIRALPAK AD (250 mm * 30 mm, 10 μm), conditions: 0.1% NH 3 . H 2 O IPA, starting with B: 45%; ending with B: 45%; flow rate (ml / min): 70)] to obtain isomer 1 (6.2 mg, yield: 9.2%) and isomer 2 (7.7 mg, yield: 11%).

[0329] Analysis method

[0330] Column: DAICEL CHIRALCEL OD-3 (100 mm * 4.6 mm, 3 μm);

[0331] Mobile phase: A: CO 2 B: ethanol (0.05% DEA), gradient: 5% to 40% of B in 4 min, maintaining 40% of B for 2.5 min, then 5% of B for 1.5 min;

[0332] Flow rate: 2.8 mL / min;

[0333] ABPR: 1500 psi;

[0334] Temperature: 35 °C.

[0335] The compound with a retention time of 3.518 min was defined as isomer 1;

[0336] MS(ESI) m / z 434.3 [M+H] +

[0337] 1 1H NMR (400 MHz, DMSO-d6) δ = 8.37 (s, 1H), 7.45 (br s, 1H), 6.90 (d, J = 11.5 Hz, 1H), 4.93 (d, J = 5.3 Hz, 1H), 4.86 (br d, J = 6.8 Hz, 1H), 4.48 (dd, J = 1.6, 11.7 Hz, 1H), 4.24 (br d, J = 9.3 Hz, 1H), 3.84 - 3.74 (m, 3H), 3.53 - 3.40 (m, 2H), 3.03 (br t, J = 10.2 Hz, 1H), 2.60 (s, 3H), 2.01 - 1.90 (m, 1H), 1.54 - 1.44 (m, 1H), 1.38 (d, J = 6.5 Hz, 3H).

[0338] The compound with a retention time of 4.165 min was defined as isomer 2;

[0339] MS (ESI) m / z 434.3 [M + H] +

[0340] 1 1H NMR (400 MHz, DMSO-d6) δ = 8.37 (s, 1H), 7.46 (br s, 1H), 6.91 (br d, J = 11.5 Hz, 1H), 4.93 (d, J = 5.3 Hz, 1H), 4.86 (br d, J = 6.5 Hz, 1H), 4.47 (br d, J = 10.5 Hz, 1H), 4.23 (br d, J = 9.8 Hz, 1H), 3.86 - 3.74 (m, 3H), 3.54 - 3.44 (m, 1H), 3.32 - 3.27 (m, 1H), 3.03 (br t, J = 10.3 Hz, 1H), 2.60 (s, 3H), 1.95 (br d, J = 13.1 Hz, 1H), 1.54 - 1.43 (m, 1H), 1.38 (d, J = 6.5 Hz, 3H).

[0341] Example 4

[0342] (3S,4R)-4-((5-chloro-4-((S)-8-fluoro-2-(2-hydroxypropan-2-yl)-3-methyl-3,4-dihydro-5-oxa-1,2a-diazacenaphthylen-6-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol isomer 1

[0343] (3S,4R)-4-((5-chloro-4-((R)-8-fluoro-2-(2-hydroxypropan-2-yl)-3-methyl-3,4-dihydro-5-oxa-1,2a-diazacyclopenta[c]acenaphthylen-6-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol isomer 2

[0344]

[0345] The first step

[0346] 6-bromo-8-fluoro-3-methyl-3,4-dihydro-5-oxa-1,2a-diazacyclopenta[c]acenaphthylene-2-carbaldehyde 4a

[0347] Under a nitrogen atmosphere, compound 1f (2.0 g, 7 mmol) and selenium dioxide (3.1 g, 28 mmol) were successively added to 30 mL of 1,4-dioxane. The reaction was carried out at 95 °C for 8 hours. The reaction solution was cooled to room temperature, filtered, the filtrate was collected, and the residue after concentration under reduced pressure was purified by C-18 reverse-phase chromatography to obtain the title compound 4a (930 mg, yield: 44%).

[0348] MS(ESI) m / z 299.1, 301.1 [M+H] +

[0349] The second step

[0350] 1-(6-bromo-8-fluoro-3-methyl-3,4-dihydro-5-oxa-1,2a-diazacyclopenta[c]acenaphthylen-2-yl)ethan-1-ol 4b

[0351] Under a nitrogen atmosphere, compound 4a (930 mg, 3.1 mmol) was dissolved in 20 mL of tetrahydrofuran. The temperature was lowered to -20 °C, and a solution of methylmagnesium bromide in tetrahydrofuran (3 mol / L, 1.5 mL, 4.5 mmol) was added dropwise, and the reaction was carried out at -20 °C for 4 hours. The reaction was quenched by adding 5 mL of water. The residue after concentration under reduced pressure of the reaction solution was purified by C-18 reverse-phase chromatography to obtain the title compound 4b (830 mg, yield: 85%).

[0352] MS(ESI) m / z 315.2, 317.2 [M+H] +

[0353] The third step

[0354] 1-(6-bromo-8-fluoro-3-methyl-3,4-dihydro-5-oxa-1,2a-diazacyclopenta[c]acenaphthylen-2-yl)ethan-1-one 4c

[0355] 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. After the reaction solution was cooled to room temperature, it was filtered, the filtrate was collected, and the residue after concentration under reduced pressure was purified by C-18 reverse-phase chromatography to obtain the title compound 4c (350 mg, yield: 59%).

[0356] MS(ESI) m / z 313.1, 315.1 [M+H] +

[0357] The fourth step

[0358] 2-(6-Bromo-8-fluoro-3-methyl-3,4-dihydro-5-oxa-1,2a-diazacenaphthylen-2-yl)propan-2-ol 4d

[0359] Under a nitrogen atmosphere, compound 4c (350 mg, 1.1 mmol) was dissolved in 20 mL of tetrahydrofuran. The temperature was lowered to -20 °C, and a solution of methylmagnesium bromide in tetrahydrofuran (3 mol / L, 0.7 mL, 2.1 mmol) was added dropwise, and the reaction was carried out at -20 °C for 4 hours. 5 mL of water was added to quench the reaction. The residue after concentration of the reaction solution under reduced pressure was purified by C-18 reverse-phase chromatography to obtain the title compound 4d (260 mg, yield: 71%).

[0360] MS(ESI) m / z 329.2, 331.2 [M+H] +

[0361] The fifth step

[0362] 2-(6-(2,5-Dichloropyrimidin-4-yl)-8-fluoro-3-methyl-3,4-dihydro-5-oxa-1,2a-diazacenaphthylen-2-yl)propan-2-ol 4e

[0363] Under a nitrogen atmosphere, compound 4d (260 mg, 0.8 mmol), bis(pinacolato)diboron (305 mg, 1.2 mmol), potassium acetate (157 mg, 1.6 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (117 mg, 0.2 mmol) were successively dissolved in 5 mL of 1,4-dioxane. The reaction was carried out at 100 °C for 2 hours. The reaction solution was cooled to room temperature, and 1,3,5,7-tetramethyl-6-phenyl-2,4,8-trioxa-6-phosphaadamantane (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. 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. After concentration under reduced pressure, the residue was purified by C-18 reverse-phase chromatography to obtain the title compound 4e (129 mg, yield: 41%).

[0364] MS(ESI) m / z 397.3 [M+H] +

[0365] The sixth step

[0366] (3S,4R)-4-((5-chloro-4-((S)-8-fluoro-2-(2-hydroxypropan-2-yl)-3-methyl-3,4-dihydro-5-oxa-1,2a-diazacenaphthylen-6-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol isomer 1

[0367] (3S,4R)-4-((5-chloro-4-((R)-8-fluoro-2-(2-hydroxypropan-2-yl)-3-methyl-3,4-dihydro-5-oxa-1,2a-diazacenaphthylen-6-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol isomer 2

[0368] 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 successively dissolved in 5 mL of tetrahydrofuran. Cesium carbonate (202 mg, 0.62 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 the crude product mixture 4f. The crude product was subjected to chiral resolution [column: DAICEL CHIRALPAK AD (250 mm * 30 mm, 10 μm), conditions: 0.1% NH3 . H 2 O ETOH, Starting B: 40%; Ending B: 40%; Flow rate (ml / min): 60) gave the title compound isomer 1 (25.2 mg, yield: 17%) and the title compound isomer 2 (22 mg, yield: 15%).

[0369] Analysis method

[0370] Column: DAICEL CHIRALCEL AD-3 (100 mm * 4.6 mm, 3 μm);

[0371] Mobile phase: A: CO 2 B: Ethanol (0.05% DEA), gradient: 5% to 40% of B in 2 min, maintaining 40% of B for 1.2 min, then 5% of B for 0.8 min;

[0372] Flow rate: 4 mL / min;

[0373] ABPR: 1500 psi;

[0374] Temperature: 35 °C.

[0375] The compound with a retention time of 1.887 min was defined as isomer 2;

[0376] MS (ESI) m / z 478.1 [M + H]+

[0377] 1 H NMR (400 MHz, DMSO-d6) δ = 8.37 (s, 1H), 7.45 (br s, 1H), 6.92 (d, J = 11.3 Hz, 1H), 5.82 (br s, 1H), 5.23 (q, J = 6.4 Hz, 1H), 4.93 (d, J = 5.3 Hz, 1H), 4.47 (d, J = 11.0 Hz, 1H), 4.22 (br d, J = 11.3 Hz, 1H), 3.89 - 3.67 (m, 3H), 3.39 - 3.25 (m, 2H), 3.03 (br t, J = 10.2 Hz, 1H), 1.95 (br d, J = 10.5 Hz, 1H), 1.68 (s, 3H), 1.62 (s, 3H), 1.54 - 1.47 (m, 1H), 1.45 (d, J = 6.5 Hz, 3H).

[0378] The compound with a retention time of 2.078 min was defined as isomer 1;

[0379] MS (ESI) m / z 478.1 [M + H] +

[0380] 1 1H NMR (400 MHz, DMSO-d6) δ = 8.37 (s, 1H), 7.45 (br s, 1H), 6.92 (d, J = 11.5 Hz, 1H), 5.82 (s, 1H), 5.29 - 5.19 (m, 1H), 4.93 (d, J = 5.5 Hz, 1H), 4.47 (d, J = 11.3 Hz, 1H), 4.21 (br d, J = 10.3 Hz, 1H), 3.85 - 3.75 (m, 3H), 3.53 - 3.40 (m, 2H), 3.03 (br t, J = 10.4 Hz, 1H), 1.94 (br s, 1H), 1.67 (s, 3H), 1.62 (s, 3H), 1.53 - 1.47 (m, 1H), 1.45 (d, J = 6.5 Hz, 3H).

[0381] Biological evaluation

[0382] The present invention will be further described and explained in combination with test examples below, but these examples are not intended to limit the scope of the present invention.

[0383] Test Example 1: Detection of Cyclin-Dependent Kinase Activity of the Published Compound

[0384] 1. Experimental Materials

[0385]

[0386] Compound A is Example A94 of WO 2019 / 207463A1 and was synthesized according to the method disclosed in this patent application.

[0387] 2. Kinase Activity Test (CDK4 / Cyclin D1, CDK6 / Cyclin D3)

[0388] The in vitro CDK kinase activity was tested by the method of Mobility Shift Assay. In the experiment, the starting concentration of the test compound for the inhibition test of CDK activity was 300 nM, diluted 3-fold, with a total of 10 concentrations, and tested in duplicate. Staurosporine was used as a standard control.

[0389] Prepare 1× kinase buffer (CDK2) (50 mM HEPES, pH 7.5, 0.0015% Brij-35), 1× 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 an appropriate amount of kinase to 1× kinase buffer to prepare a 2.5× enzyme solution; prepare a 5× compound dilution corresponding to the test concentration of the compound (1× kinase buffer, 10% DMSO); add an appropriate amount of FAM-labeled polypeptide and ATP to 1× kinase buffer to prepare a 2.5× substrate solution. Add 5 μl of 5× compound dilution and 10 μl of 2.5× enzyme solution to the reaction wells of a 384-well reaction plate, mix well, and incubate at room temperature for 10 minutes; then add 10 μl of 2.5× substrate solution to the 384-well plate and centrifuge at 1000 rpm for 1 minute; incubate the reaction plate at 28 °C for 60 minutes (biochemical incubator model: SPX-100B-Z); add 30 μl of stop solution to the 384-well reaction plate to terminate the reaction and centrifuge at 1000 rpm for 1 minute; finally, read the conversion rate data on a Caliper EZ Reader Ⅱ (excitation wavelength: 400 nm, emission wavelengths: 445 nm and 520 nm).

[0390] The IC 50 value of the compound was fitted using the XLFit excel add-in version 5.4.0.8. Fitting formula:

[0391] Y = Bottom + (Top - Bottom) / (1 + 10^((LogIC50 - X)*HillSlope)).

[0392] X: logarithm value of the compound concentration; Y: percentage inhibition of the compound.

[0393] 3. Kinase activity assay (CDK1 / cyclin B, CDK9 / cyclin T1)

[0394] The starting concentration for the in vitro CDK (CDK2, CDK9) kinase activity assay was 1 μM, diluted 3-fold, with a total of 10 concentrations, and tested in duplicate. The compound PHA-793887 was used as a control compound.

[0395] Prepare 1x kinase reaction buffer (40 mM Tris-HCl, pH 7.4, 20 mM Mg2Cl2, 0.1 mg / ml BSA, 50 μM DTT), 1 volume of 5x kinase reaction buffer and 4 volumes of water, and add DTT (final concentration 50 μM). Transfer 50 nL of the diluted compound working solution (final DMSO concentration is 1%) to each well of the reaction plate (784075, Greiner) using an Echo 655. Seal the reaction plate with a sealing film and centrifuge at 1000 g for 1 minute. Prepare 2x enzyme (0.3 ng / uL CDK2 / cyclin E1 or CDK9 / cyclin T1) with 1x kinase reaction buffer, add 2.5 uL of the above kinase solution to each well, seal the reaction plate with a sealing film, centrifuge at 1000 g for 1 minute, and incubate at room temperature for 10 minutes. Prepare 2x kinase substrate and ATP mixture with 1× kinase reaction buffer, and the 2× CDK2 / CylinE1 kinase substrate is 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, centrifuge at 1000 g for 30 seconds, and start the reaction. After the kinase test reacts at room temperature for 120 minutes, add 4 μl of ADP-Glo reagent and react at room temperature for 40 minutes. Then add 8 μl of kinase detection reagent and react at room temperature for 40 minutes, and read the luminescence signal using an Envision 2104. Data analysis is as follows

[0396] a) Inhibition percentage: % Inhibition = 100 - (Signalcmpd - SignalAve_PC) / (SignalAve_VC - SignalAve_PC) × 100.

[0397] SignalAve_PC: The average value of all positive control wells on the entire plate.

[0398] SignalAve_VC: The average value of all negative control wells on the entire plate.

[0399] Signalcmpd: The average value of the wells corresponding to the test compound.

[0400] b) Compound IC 50 : Calculate using the following non-linear fitting formula with GraphPad 8.0.

[0401] Y = Bottom + (Top - Bottom) / (1 + 10^((LogIC50 - X)*HillSlope))

[0402] X: Log value of the compound concentration; Y: Inhibition percentage of the compound.

[0403] The CDK (CDK1, CDK4, CDK6, CDK9) kinase biochemical inhibitory activities of the compounds of the present disclosure were determined by the above tests, and the measured IC 50 values are shown in Tables 1, 2, and 3.

[0404] Table 1.

[0405]

[0406]

[0407] Table 2.

[0408]

[0409] Table 3.

[0410]

[0411] Test Example 2, CYP Inhibition Experiment

[0412] The metabolic reactions of representative substrates of the five major human CYP subtypes (CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP3A4) were evaluated using 150-donor pooled human liver microsomes (purchased from Corning, catalog number 452117). The effects of the test compounds at different concentrations 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).

[0413] A reaction system of 200 μL (100 mmol / L phosphate buffer, pH 7.4, containing DMSO at a volume ratio of 0.3%, acetonitrile at a volume ratio of 0.6%, and methanol at a volume ratio of 0.1%) of 30 μM phenacetin, 10 μM diclofenac sodium, 35 μM S-mephenytoin, 5 μM bufuralol hydrochloride, 3 μM midazolam, 1 mM NADPH, the test compounds (at concentrations of 0.1, 0.3, 1, 3, 10, 30 μmol / L respectively), or the positive compound or the blank control and pooled human liver microsomes (0.2 mg / mL) was incubated at 37 °C for 5 minutes. 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. It was cooled on ice for 20 minutes and then centrifuged at 4000 rpm for 20 minutes to precipitate proteins. 200 μL of the supernatant was taken for LC / MS / MS analysis.

[0414] The peak areas were calculated based on the chromatograms. The residual activity ratio (%) was calculated using the following formula:

[0415] Peak area ratio = metabolite peak area / internal standard peak area

[0416] Residual activity ratio (%) = peak area ratio of the test compound group / peak area ratio of the blank group

[0417] Half inhibitory concentration (IC 50 ) was calculated by Excel XLfit 5.3.1.3

[0418] The measured CYP half inhibitory concentration (IC 50 ) values are shown in Table 4 below

[0419] Table 4. Half inhibitory concentration (IC 50 ) of the compounds of the present disclosure against CYP

[0420]

[0421] Test Example 3. Solubility test experiment

[0422] The thermokinetic solubility of the compound in phosphate buffer at pH 7.4 was determined. The sample supernatant and the standard with known concentration were both detected by LC / MS / MS

[0423] 1. Materials and reagents

[0424] Compound A (Compound A is Example A94 of WO 2019207463A and was synthesized by referring to the method disclosed in this patent application)

[0425] NaH 2 PO 4 ·2H 2 O (analytical pure), NaH 2 PO 4 (analytical pure), NaOH (analytical pure)

[0426] 1.5 mL flat-bottom glass tube (BioTech Solutions); molded polytetrafluoroethylene lid (BioTech

[0427] Solutions), polytetrafluoroethylene-coated stir bar (BioTech Solutions), Eppendorf Comfort Thermomixer and 96-well deep-well plate

[0428] 2. Preparation of 0.01 M sodium phosphate buffer at pH 7.4

[0429] Weigh 15.6 g of NaH 2 PO 4 ·2H 2Put O into a 1L glass bottle and dissolve it with 1L of deionized water. The pH of the solution is approximately 4.7, and then adjust the pH value to 7.4 with 10M NaOH.

[0430] 3. Solubility determination process

[0431] Accurately weigh 1mg of powder of each compound and put it into a glass tube. Add phosphate buffer solution with pH 7.4 to the above glass tube, and the addition amount is 1mL per milligram. Add a stir bar to each glass tube, then cover the lid. Put the sample tray with the glass tubes into an Eppendorf Comfort Thermomixer, incubate at 25°C and 1100rpm for 24 hours. After the incubation, open the lid, suck out the stir bar with a magnet, and record the phenomena in each glass tube. Centrifuge the plate at 25°C and 4000rpm for 30 minutes. Pipette 750μL of the supernatant. Wash the pipette tip with acetonitrile for 5 seconds, and then wash it with pure water for 5 seconds. Then discharge the first 50μL of waste liquid, and add the remaining 700μL to another 96-well sample tray with glass tubes, and centrifuge for 30 minutes (25°C, 4000rpm). Pipette 10μL of the sample after the second centrifugation into 990μL of a mixture of acetonitrile and water (1:1) containing internal standard (100 times the sample). Pipette 10μL of the diluted solution into 990μL of a mixture of acetonitrile and water (1:1) containing internal standard (10,000 times the sample). The sample dilution factor may vary according to the solubility value and LC / MS signal response.

[0432] Table 5. Recording of phenomena and dilution factor

[0433]

[0434] 4. Preparation of standards

[0435] Accurately weigh 1mg of compound powder and add it to each glass tube. Add DMSO to each glass tube, and the addition amount is 1mL per milligram. Add a stir bar to each glass tube, then cover the lid. Put the tray with the standard glass tubes into an Eppendorf Comfort Thermomixer, incubate at 25°C and 1100 revolutions for 2 hours to fully dissolve the powder. Observe whether the solid is completely dissolved, and record the compounds that cannot be completely dissolved in the DMSO solution. Pipette 10μL of the 1mg / mL standard into 990μL of a mixture of acetonitrile and water (1:1) containing internal standard to obtain a 10μg / mL standard. Pipette 10μL of the 10μg / mL standard into 990μL of a mixture of acetonitrile and water (1:1) containing internal standard to obtain a 0.1μg / mL standard. The sample dilution factor may vary according to the LC / MS signal response. The samples are analyzed by LC / MS / MS. All compounds are tested individually.

[0436] 5. Data Calculation

[0437] All calculations were performed using Microsoft Excel.

[0438] The samples were analyzed by LC / MS / MS and quantified according to the standards of known concentrations. The solubility of the compound to be tested was calculated using the following formula: [Sample] = Area Ratio of Sample × DF of Sample × [STD] / Area Ratio of STD

[0439] DF: Dilution Factor.

[0440] STD: Standard of the compound to be tested

[0441] Table 6. Solubility of the Compounds of the Present Disclosure

[0442] 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

[0443] Test Example 4. PXR Induction Experiment

[0444] 1. Evaluate the potential of the compound to be tested to induce the activity of drug-metabolizing enzymes by activating PXR in vitro. The EC 50 values of the test compound at different concentrations (30, 10, 3.33, 1.11, 0.370, and 0.123 μM) were obtained by activating PXR in vitro. The concentrations of the positive control rifampicin were 20, 5, 1.25, 0.312, 0.0781, and 0.195 μM.

[0445] 2. Materials and Reagents

[0446] 1) DPX2 (human PXR gene and fluorescent reporter gene stably transfected into HepG2 cells) cells were purchased from Puracyp (Carlsbad, CA).

[0447] 2) The compound to be tested was provided by the client, and the control drug (rifampicin) was purchased from Sigma (St. Louis, MO).

[0448] 3) CellTiter-Fluor TM Cell viability detection kit and One-Glo fluorescence detection kit were purchased from Promega (Madison, WI), fetal bovine serum (FBS) was purchased from Corning (Manassas, VA), 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.

[0449] 3. Experimental Procedures

[0450] 3.1 Preparation of Seeding

[0451] 1) Add 50 mL of FBS to 450 mL of cell culture medium.

[0452] 2) Culture DPX2 cells in a T-75 culture flask in an incubator at 37 °C, 5% CO 2 , with a relative humidity of 95%. Digest the cells when they reach 80 - 90% confluence at the bottom of the culture flask.

[0453] 3) Wash the surface of the cells cultured in the T-75 flask with 8 mL of PBS, aspirate 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. Then add 10 mL of excess serum-containing medium to neutralize the trypsin.

[0454] 4) Transfer the cell suspension to a conical-bottom centrifuge tube and centrifuge at 120 g for 10 minutes. Suspend the cells with seeding medium and adjust the concentration to 4x10 5 cells / mL. Add 100 μL of the diluted cells to each well of a 96-well cell culture plate. Place the culture plate in the incubator and incubate at 37 °C for 24 hours, then prepare for the PXR activation experiment.

[0455] 3.2 Drug Treatment

[0456] 1) Prepare the test compound and the positive compound (rifampicin) with DMSO, and dilute the compounds with serum-free medium at 37 °C. The final concentrations of the positive control rifampicin are 20, 5, 1.25, 0.312, 0.0781, and 0.195 μM, and the final concentrations of the test compounds are 30, 10, 3.33, 1.11, 0.370, and 0.123 μM. The final concentration of DMSO is 0.1%. Add 1 μL of DMSO to 1 mL of pre-incubated medium as the solvent control group.

[0457] 2) Take out the cell culture plate from the incubator, discard the medium, add 100 μL of the test compound and the positive compound to the appropriate wells, with two parallels in each group. Place the cell plate in the incubator and incubate for 24 hours.

[0458] 3.3 Quantitative Determination of PXR Activation

[0459] 1) Two days after drug treatment, the culture can be used for quantitative detection of PXR activation.

[0460] 2) Add CellTiter-Fluor TMThe cell viability detection kit and One-Glo luciferase reagent were equilibrated to room temperature. The GF-AFC (10 μL) substrate was added to the detection buffer (10 mL) to form a 2X reagent, which was then diluted to 1X with 10 mL of PBS; the ONE-Glo substrate was added to the ONE-Glo luciferase detection buffer.

[0461] 3) Remove the cell culture plate from the incubator, discard the medium in each well, wash twice with PBS, add the 1X CellTiter-Fluor TM reagent to a sterilized pipette tip box, and use a multichannel pipette to aspirate 50 μL and add it to each well, then incubate at 37 °C for 30 minutes.

[0462] 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.

[0463] 5) Pour the ONE-Glo reagent into the pipette tip box, then use a multichannel pipette to aspirate 50 μL and add it to each well, gently mix the reagent, and incubate at room temperature for 5 minutes with mixing. After incubation, use a luminometer to read the luminescence value of each well.

[0464] 4. Calculation of cell induction value

[0465] 4.1 Cell viability

[0466] Cell viability calculation formula:

[0467] Percentage of cell viability (%) = I (sample) / (I (vector) x 100

[0468] I (sample) is the fluorescence intensity of the sample, and I (vector) is the fluorescence intensity of 0.1% DMSO on the cells.

[0469] 4.2 Calculation of cell induction value

[0470] All data were calculated using Microsoft Excel.

[0471] The activity of luciferase was represented by RFU / RLU. RLU is the average luminescence intensity value of two parallels for each compound at each concentration, and RFU is the average fluorescence intensity of two parallels for each compound at each concentration. Induction fold calculation formula:

[0472]

[0473] Table 7. Part of the measured PXR induction values

[0474]

[0475]

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof, which is a compound of formula I-3 or a pharmaceutically acceptable salt thereof, wherein R 9 is OH or NH 2 and said NH 2 is optionally substituted by 1 or 2 R a’ or R a” and R a’ and R a” are each independently C 1-6 alkyl; R 10 Each independently selected from OH, halogen, CN, NH 2 , 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, wherein the C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, and 3-12 membered heterocycloalkyl are each independently optionally substituted with one or more R b or deuterium; 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.

2. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, wherein R 9 is OH or NH 2 and said NH 2 is optionally substituted by 1 or 2 R a’ or R a” ; R a’ and R a” are each independently 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, wherein the C 1-6 alkyl and C 1-6 alkoxy are each independently optionally substituted with one or more R b or deuterium, and 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 3. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 2, wherein R 9 is OH; m is 0; R 7 is F or Cl; R 3 is F or Cl; R 4 is H or C 1-6 alkyl, said C 1-6 alkyl is optionally substituted by one or more R b or deuterium, and R b is selected from OH, CN, and halogen; R 5 and R 6 are each independently H or C 1-6 alkyl group.

4. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 2, wherein R 9 is OH; m is 0; R 7 is F or Cl; R 3 is F or Cl; R 4 is H or C 1-6 alkyl, said C 1-6 alkyl is optionally substituted by one or more R b or deuterium, R b is OH; R 5 and R 6 are each independently selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and tert-butyl.

5. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 2, wherein R 9 is OH; m is 0; R 7 is F or Cl; R 3 is F or Cl; R 4 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 substituted with one or more R b substituents; R b is OH; R 5 and R 6 are each independently selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and tert-butyl.

6. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 2, wherein R 9 is OH; m is 0; R 7 is F or Cl; R 3 is F or Cl; R 4 selected from H, methyl, ethyl, n-propyl and isopropyl; R 5 and R 6 are each independently selected from H, methyl, ethyl, n-propyl, and isopropyl.

7. A compound of formula I or a pharmaceutically acceptable salt thereof according to claim 2, which is selected from the following compounds or pharmaceutically acceptable salts thereof, 8. An isotopically substituted compound of the compound of formula I or a pharmaceutically acceptable salt thereof according to any one of claims 1-7, wherein the isotopically substituted compound is a deuterium atom substituted compound.

9. A method for preparing the compound of formula I or a pharmaceutically acceptable salt thereof according to any one of claims 1-7, comprising the step of reacting a compound of formula I-B with a compound of formula I-C to form a compound of formula I, wherein, LG 1 is a leaving group, and the leaving group is selected from halogen, sulfonate, boric acid and borate; X, Y, Z, L, R 1 , R 2 , R 3 , R 4 , R 5 and R 6 are defined as in claim 1.

10. A pharmaceutical composition comprising the compound of formula I or a pharmaceutically acceptable salt thereof according to any one of claims 1-7, or the isotopically substituted compound according to claim 8, and a pharmaceutically acceptable excipient.

11. Use of the compound of formula I or a pharmaceutically acceptable salt thereof according to any one of claims 1-7, or the isotopically substituted compound according to claim 8, or the pharmaceutical composition according to claim 10 in the preparation of a drug for preventing and / or treating a cyclin-dependent kinase related disease.

12. Use of the compound of formula I or a pharmaceutically acceptable salt thereof according to any one of claims 1-7, or the isotopically substituted compound according to claim 8, or the pharmaceutical composition according to claim 10 in the preparation of a drug 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.

13. A compound of formula I-B or a pharmaceutically acceptable salt thereof, wherein, LG 1 is a leaving group, and the leaving group is selected from halogen, sulfonate, boric acid and borate; X, Y, Z, L, R 3 , R 4 , R 5 and R 6 as defined in claim 1.

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