WO2023083201A1 - Aminopyrazole derivative, and preparation method therefor and use thereof - Google Patents

Abstract

Provided in the present disclosure are an aminopyrazole derivative, and a preparation method therefor and the use thereof. Specifically, provided in the present disclosure is a compound as represented by formula I or a pharmaceutically acceptable salt thereof. The compound as represented by formula I can be used as a cyclin-dependent kinase inhibitor, and used for preventing and/or treating a disease related to a protein-dependent kinase or cyclin-dependent protein. Each group in general formula I is as defined in the description.

Description

A kind of aminopyrazole derivative and its preparation method and application technical field

The disclosure relates to an aminopyrazole derivative, a preparation method and application thereof, and belongs to the field of medicine.

Background technique

Cyclin-dependent kinases (CDKs) are members of the serine/threonine kinase subfamily, and each CDK/cyclin complex is responsible for a specific phase transition or progression within the cell cycle, which plays an important role in regulating eukaryotic cell division and important role in proliferation. Cyclin-dependent kinase catalytic units are activated by regulatory subunits called cyclins. At least 16 mammalian cell 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).

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

Overexpression of CDK2 is associated with abnormal regulation of the cell cycle. The cyclin E/CDK2 complex plays an important role in regulating the G1/S transition, histone biosynthesis and centrosome duplication. Progressive phosphorylation of Rb by cyclin D/Cdk4/6 and cyclin E/Cdk2 releases the G1 transcription factor E2F and promotes S phase entry. Activation of cyclin A/CDK2 during early S phase promotes phosphorylation of endogenous substrates that allow DNA replication and inactivation of E2F to complete S phase (Nat.Rev.Drug.Discov.2015; 14( 2): 130-146). Cyclin E is overexpressed in a variety of cancers, especially breast cancer, lung cancer, leukemia, and lymphoma (Guo Cuiping et al., Cyclin E regulation and malignant tumors. International Journal of Oncology, 2012,39(005):337 -340), the amplification or overexpression of cyclin E is also associated with poor prognosis in ovarian, gastric, endometrial and other cancers.

Studies have shown that inhibition of CDK2 kinase induces tumor cell apoptosis, but causes only minor damage to normal cells. The monomeric form of the CDK kinase is inactive, whereas the binding of cyclin A/E to CDK2 and triggering phosphorylation activates CDK2. CDK2 also binds cyclin A for progression through S phase and participates in DNA repair. In recent years, major companies have identified and discovered a series of selective CDK 2 inhibitors for the treatment of cancer and other diseases, such as Seliciclib, Dinaciclib, etc., but in order to achieve better cancer treatment effects, better meet Market demand still requires the development of a new generation of selective CDK2 inhibitors with high efficiency and low toxicity.

Contents of the invention

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

Wherein, said ^R1 and ^R2 are each independently selected from hydrogen, alkyl, haloalkyl, hydroxyalkyl, cycloalkyl or heterocycloalkyl, said alkyl, haloalkyl, hydroxyalkyl, ring Alkyl or heterocycloalkyl is optionally substituted by one or more substituents independently selected from R ^1A ;

The R ^1A is selected from halogen, hydroxyl, alkyl, alkoxy, haloalkyl, haloalkoxy, heterocycloalkyl or NR'(R"), and the alkyl, alkoxy, haloalkyl, Haloalkoxy or heterocycloalkyl is optionally substituted by one or more substituents independently selected from R ⁵ ;

Or R ¹ and R ² form a 3- to 7-membered heterocycloalkyl group with the nitrogen atom to which they are jointly connected, and the heterocycloalkyl group is optionally substituted by one or more substituents independently selected from R ^1B ;

The R ^1B is selected from independently halogen, cyano, hydroxyl, amino, alkyl or alkoxy;

The R ³ is selected from H, COR' or

The ring A is selected from 5 to 6 membered aryl, 5 to 6 membered heteroaryl, cycloalkyl or heterocycloalkyl;

或氧代基，所述的烯基、炔基、烷基、烷氧基、卤代烷基、卤代烷氧基、羟烷基、环烷基、杂环烷基、环烷基氧基、杂环烷基氧基、芳基、杂芳基任选被一个或多个各自独立地选自R ^4A的取代基取代； The ^R4 are each independently selected from halogen, cyano, nitro, hydroxyl, alkenyl, alkynyl, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclic Alkyl, cycloalkyloxy, heterocycloalkyloxy, aryl, heteroaryl, SR', SOR', SO ₂ R', -NHSO ₂ R', SO ₂ NR'(R"), NR '(R"), COR', -NHCOR', COOR', CONR'(R"), -(P=O)R'(R"),

Or oxo, the alkenyl, alkynyl, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocycloalkyl, cycloalkyloxy, heterocycloalkane Oxygen, aryl, heteroaryl are optionally substituted by one or more substituents independently selected from R ^4A ;

Alternatively, two adjacent R ⁴ together form ring A optionally substituted by one or more independently selected from R ^4A

所述的烷基、烷氧基、卤代烷基、卤代烷氧基、羟烷基、环烷基、杂环烷基、环烷基氧基、杂环烷基氧基、芳基、杂芳基任选被一个或者多个R ⁸取代； Described R ^4A is selected from halogen, cyano, nitro, hydroxyl, alkenyl, alkynyl, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocycloalkyl, Cycloalkyloxy, heterocycloalkyloxy, aryl, heteroaryl, SR', SOR', SO ₂ R', -NHSO ₂ R', SO ₂ NR'(R"), NR'(R "), COR', -NHCOR', COOR', CONR'(R"), -(P=O)R'(R"), or

The alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocycloalkyl, cycloalkyloxy, heterocycloalkyloxy, aryl, heteroaryl are any is selected to be replaced by one or more R ⁸ ;

Said X is independently selected from O or S;

The R ⁵ is selected from halogen, hydroxyl, alkyl, cyano, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl or NR'(R");

The R ^and R ^are each independently selected from hydrogen, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl or heterocycloalkyl;

The ^R is selected from halogen, hydroxyl, cyano, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl or NR'(R");

The R' and R" are each independently selected from H or alkyl;

Said n is selected from 1, 2, 3, 4, 5.

In an optional embodiment, the compound represented by formula I or a pharmaceutically acceptable salt thereof in the present disclosure is a compound represented by formula II or a pharmaceutically acceptable salt thereof,

Wherein, the R ¹ , R ² , R ³ , R ⁴ , and n are respectively as defined in the compound represented by formula I.

In an optional embodiment, the compound represented by formula I or a pharmaceutically acceptable salt thereof in the present disclosure is a compound represented by formula III or a pharmaceutically acceptable salt thereof,

Wherein, the R ¹ , R ² , R ⁴ , and n are respectively as defined in the compound represented by formula I.

In an optional embodiment, the compound represented by formula I or a pharmaceutically acceptable salt thereof in the present disclosure is a compound represented by formula IV or a pharmaceutically acceptable salt thereof,

Wherein, the R ¹ , R ² , R ⁴ , and n are respectively as defined in the compound represented by formula I.

In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein ring A is selected from phenyl, five-membered nitrogen-containing heteroaryl, six-membered nitrogen-containing heteroaryl.

In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein ring A is selected from pyridyl, pyrimidinyl, piperidinyl, imidazolyl, Pyrrolyl, pyrazolyl.

In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, IV or pharmaceutically acceptable salts thereof, wherein ring A is selected from pyridyl.

In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, IV or pharmaceutically acceptable salts thereof, wherein ring A is selected from phenyl.

所述G ¹、G ²、G ³、G ⁴和G ⁵各自独立地选自CR ⁴或N，其中，G ¹、G ²、G ³、G ⁴和G ⁵中至少有一个为CR ⁴，所述R ⁴如式I所示化合物中定义。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, IV or pharmaceutically acceptable salts thereof, wherein Ring A is I-1,

The G ¹ , G ² , G ³ , G ⁴ and G ⁵ are each independently selected from CR ⁴ or N, wherein at least one of G ¹ , G ² , G ³ , G ⁴ and G ⁵ is CR ⁴ , The R ⁴ is as defined in the compound shown in formula I.

所述G ¹、G ²、G ³和G ⁴各自独立地选自CR ⁴或N或NR ⁴，所述R ⁴如式I所示化合物中定义。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, IV or pharmaceutically acceptable salts thereof, wherein Ring A is I-2,

The G ¹ , G ² , G ³ and G ⁴ are each independently selected from CR ⁴ or N or NR ⁴ , and the R ⁴ is as defined in the compound shown in formula I.

所述G ²、G ³、G ⁴和G ⁵各自独立地选自CR ⁴，所述R ⁴如式I所示化合物中定义。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

The G ² , G ³ , G ⁴ and G ⁵ are each independently selected from CR ⁴ , and the R ⁴ is as defined in the compound shown in formula I.

所述G ¹、G ³、G ⁴和G ⁵各自独立地选自CR ⁴，所 述R ⁴如式I所示化合物中定义。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

The G ¹ , G ³ , G ⁴ and G ⁵ are each independently selected from CR ⁴ , and the R ⁴ is as defined in the compound represented by formula I.

所述G ¹、G ²、G ⁴和G ⁵各自独立地选自CR ⁴，所述R ⁴如式I所示化合物中定义。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

The G ¹ , G ² , G ⁴ and G ⁵ are each independently selected from CR ⁴ , and the R ⁴ is as defined in the compound shown in formula I.

所述G ³、G ⁴和G ⁵各自独立地选自CR ⁴，所述R ⁴如式I所示化合物中定义。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

The G ³ , G ⁴ and G ⁵ are each independently selected from CR ⁴ , and the R ⁴ is as defined in the compound shown in formula I.

所述G ²、G ⁴和G ⁵各自独立地选自CR ⁴，所述R ⁴如式I所示化合物中定义。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

The G ² , G ⁴ and G ⁵ are each independently selected from CR ⁴ , and the R ⁴ is as defined in the compound shown in formula I.

所述G ²、G ³和G ⁴各自独立地选自CR ⁴，所述R ⁴如式I所示化合物中定义。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

The G ² , G ³ and G ⁴ are each independently selected from CR ⁴ , and the R ⁴ is as defined in the compound shown in formula I.

所述G ¹、G ³和G ⁴各自独立地选自CR ⁴，所述R ⁴如式I所示化合物中定义。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

The G ¹ , G ³ and G ⁴ are each independently selected from CR ⁴ , and the R ⁴ is as defined in the compound shown in formula I.

所述G ²、G ³和G ⁵各自独立地选自CR ⁴，所述R ⁴如式I所示化合物中定义。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

The G ² , G ³ and G ⁵ are each independently selected from CR ⁴ , and the R ⁴ is as defined in the compound shown in formula I.

所述G ¹、G ³和G ⁵各自独立地选自CR ⁴，所述R ⁴如式I所示化合物中定义。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

The G ¹ , G ³ and G ⁵ are each independently selected from CR ⁴ , and the R ⁴ is as defined in the compound shown in formula I.

所述G ³和G ⁴各自独立地选自CR ⁴，所述R ⁴如式I所示化合物中定义。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

The G ³ and G ⁴ are each independently selected from CR ⁴ , and the R ⁴ is as defined in the compound shown in formula I.

所述G ⁴为CR ⁴，所述R ⁴如式I所示化合物中定义。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

The G ⁴ is CR ⁴ , and the R ⁴ is as defined in the compound shown in formula I.

所述G ³为CR ⁴，所述R ⁴如式I所示化合物中定义。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

The G ³ is CR ⁴ , and the R ⁴ is as defined in the compound shown in formula I.

In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, IV or pharmaceutically acceptable salts thereof, wherein G ¹ , G ² , G ³ , G ⁴ and G ⁵ are each independently selected from ^CR4 .

G ¹、G ²、G ³、G ⁴和G ⁵选自CR ⁴，所述

选自

In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

G ¹ , G ² , G ³ , G ⁴ and G ⁵ are selected from CR ⁴ , the

selected from

为

In an optional embodiment, the present disclosure provides a compound represented by formula I, II, III, IV or a pharmaceutically acceptable salt thereof, the

for

G ²、G ³、G ⁴和G ⁵各自独立地选自CR ⁴。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

G ² , G ³ , G ⁴ and G ⁵ are each independently selected from CR ⁴ .

所述

选自

In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

said

selected from

In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

G ¹、G ³、G ⁴和G ⁵各自独立地选自CR ⁴。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

G ¹ , G ³ , G ⁴ and G ⁵ are each independently selected from CR ⁴ .

In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

G ²、G ⁴和G ⁵各自独立地选自CR ⁴，所述R ⁴如式I所示化合物中定义。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

G ² , G ⁴ and G ⁵ are each independently selected from CR ⁴ , and R ⁴ is as defined in the compound represented by formula I.

G ³、G ⁴和G ⁵各自独立地选自CR ⁴。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

G ³ , G ⁴ and G ⁵ are each independently selected from CR ⁴ .

In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

G ²、G ⁴和G ⁵各自独立地选自CR ⁴，所述R ⁴如式I所示化合物中定义。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

G ² , G ⁴ and G ⁵ are each independently selected from CR ⁴ , and R ⁴ is as defined in the compound represented by formula I.

G ²、G ³、G ⁴各自独立地选自CR ⁴，所述R ⁴如式I所示化合物中定义。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

G ² , G ³ , and G ⁴ are each independently selected from CR ⁴ , and R ⁴ is as defined in the compound shown in formula I.

G ¹、G ³、G ⁴各自独立地选自CR ⁴。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

G ¹ , G ³ , and G ⁴ are each independently selected from CR ⁴ .

In an optional embodiment, the present disclosure provides a compound represented by formula I, II, III, IV or a pharmaceutically acceptable salt thereof, wherein ring A is selected from

G ²、G ³、G ⁵各自独立地选自CR ⁴，所述R ⁴如式I所示化合物中定义。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

G ² , G ³ , and G ⁵ are each independently selected from CR ⁴ , and R ⁴ is as defined in the compound shown in formula I.

G ¹、G ³和G ⁵各自独立地选自CR ⁴，所述R ⁴如式I所示化合物中定义。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

G ¹ , G ³ and G ⁵ are each independently selected from CR ⁴ , and R ⁴ is as defined in the compound represented by formula I.

G ³、G ⁴各自独立地选自CR ⁴，所述R ⁴如式I所 示化合物中定义。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

G ³ and G ⁴ are each independently selected from CR ⁴ , and R ⁴ is as defined in the compound shown in formula I.

G ⁴选自CR ⁴，所述R ⁴如式I所示化合物中定义。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

G ⁴ is selected from CR ⁴ , and R ⁴ is as defined in the compound shown in formula I.

G ³选自CR ⁴，所述R ⁴如式I所示化合物中定义。 In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein Ring A is

G ³ is selected from CR ⁴ , and R ⁴ is as defined in the compound shown in formula I.

其中环A的定义如本文任一方案所述。 In an alternative embodiment, two adjacent R ⁴ together with ring A form an optionally substituted by one or more R ^4A

Wherein ring A is defined as described in any scheme herein.

In an optional embodiment, the present disclosure provides a compound of formula I, II, III, IV or a pharmaceutically acceptable salt thereof, wherein said ^R is selected from hydrogen, alkyl, haloalkyl, hydroxyalkane group, cycloalkyl or heterocycloalkyl.

In an optional embodiment, the present disclosure provides a compound of formula I, II, III, IV or a pharmaceutically acceptable salt thereof, wherein, said R ¹ is selected from hydrogen, C _1-6 alkyl, C _1-6 haloalkyl, C _1-6 hydroxyalkyl, 3-7 membered cycloalkyl or 3-7 membered heterocycloalkyl.

In an optional embodiment, the present disclosure provides a compound represented by formula I, II, III, IV or a pharmaceutically acceptable salt thereof, wherein said R ¹ is selected from hydrogen, C _1-3 alkyl or C _1-3 haloalkyl.

In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, IV or pharmaceutically acceptable salts thereof, wherein said R ¹ is hydrogen.

In an optional embodiment, the present disclosure provides a compound of formula I, II, III, IV or a pharmaceutically acceptable salt thereof, wherein, said R ² is selected from the group consisting of alkyl, haloalkyl, hydroxyalkyl, Cycloalkyl or heterocycloalkyl, said alkyl, haloalkyl, hydroxyalkyl, cycloalkyl or heterocycloalkyl are optionally substituted by one or more R ^1A , said R ^1A is selected from haloalkane group, alkoxy, haloalkyl, haloalkoxy, heterocycloalkyl or NR'(R"), the said alkyl, alkoxy, haloalkyl, haloalkoxy, heterocycloalkyl are optionally replaced by one Or multiple R ⁵ substitutions.

In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, IV or pharmaceutically acceptable salts thereof, wherein said R ² is selected from C _1-6 alkyl, C _{1- 6} haloalkyl, C _1-6 hydroxyalkyl, 3 to 7 membered cycloalkyl or 3 to 7 membered heterocycloalkyl, said C _1-6 alkyl, C _1-6 haloalkyl, C _{1 -6} hydroxyalkyl, 3 to 7-membered cycloalkyl or 3 to 7-membered heterocycloalkyl are optionally substituted by one or more R ^1A ; said R ^1A is selected from halogen, C _1-6 alkyl .

In an optional embodiment, the present disclosure provides a compound represented by formula I, II, III, IV or a pharmaceutically acceptable salt thereof, wherein said R ² is selected from methyl, ethyl, isopropyl, Tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, described cyclopropyl, cyclobutyl, cyclopentyl are substituted by one or more R ^1A , described R ^1A is selected from methyl, ethyl base, isopropyl.

In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, IV or pharmaceutically acceptable salts thereof, wherein said R ² is selected from isopropyl or methylcyclopropyl.

其中X为O，所述R ⁶和R ⁷各自独立地选自氢、C _1-6烷基。 In an optional embodiment, the present disclosure provides compounds represented by formulas I and II or pharmaceutically acceptable salts thereof, wherein said ^R is selected from

Wherein X is O, the R ⁶ and R ⁷ are each independently selected from hydrogen, C _1-6 alkyl.

其中X为O，所述R ⁶和R ⁷各自独立地选自氢、甲基、乙基。 In an optional embodiment, the present disclosure provides compounds represented by formulas I and II or pharmaceutically acceptable salts thereof, wherein said ^R is selected from

Wherein X is O, and the R ⁶ and R ⁷ are each independently selected from hydrogen, methyl, ethyl.

In an optional embodiment, the present disclosure provides compounds represented by formulas I and II or pharmaceutically acceptable salts thereof, wherein said ^R is selected from

所述的烯基、炔基、烷基、烷氧基、卤代烷基、卤代烷氧基、羟烷基、环烷基、杂环烷基、环烷基氧基、杂环烷基氧基、芳基、杂芳基任选被一个或多个R ^4A取代，其中，R ^4A如式I所示的化合物中定义。 In an optional embodiment, the present disclosure provides a compound of formula I, II, III, IV or a pharmaceutically acceptable salt thereof, wherein R ⁴ is selected from halogen, cyano, nitro, hydroxyl, alkenyl, Alkynyl, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocycloalkyl, cycloalkyloxy, heterocycloalkyloxy, aryl, heteroaryl, SR', SOR', SO ₂ R', -NHSO ₂ R', SO ₂ NR'(R"), NR'(R"), COR', -NHCOR', COOR', CONR'(R"), -(P=O)R'(R") or

The alkenyl, alkynyl, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocycloalkyl, cycloalkyloxy, heterocycloalkyloxy, aromatic The radical, heteroaryl is optionally substituted by one or more R ^4A , wherein R ^4A is as defined in the compound shown in formula I.

或氧代基，所述的烯基、炔基、烷基、烷氧基、卤代烷基、卤代烷氧基、羟烷基、环烷基、杂环烷基、环烷基氧基、杂环烷基氧基、芳基、杂芳基任选被一个或多个R ^4A取代，其中，R ^4A如式I所示的化合物中定义。 In an optional embodiment, the present disclosure provides a compound of formula I, II, III, IV or a pharmaceutically acceptable salt thereof, wherein R ⁴ is selected from halogen, cyano, nitro, hydroxyl, alkenyl, Alkynyl, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocycloalkyl, cycloalkyloxy, heterocycloalkyloxy, aryl, heteroaryl, SR', SOR', SO ₂ R', -NHSO ₂ R', SO ₂ NR'(R"), NR'(R"), COR', -NHCOR', COOR', CONR'(R"), -(P=O)R'(R"),

Or oxo, the alkenyl, alkynyl, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocycloalkyl, cycloalkyloxy, heterocycloalkane Baseoxy, aryl, heteroaryl are optionally substituted by one or more R ^4A , wherein R ^4A is as defined in the compound shown in formula I.

In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, IV or pharmaceutically acceptable salts thereof, wherein R ⁴ is oxo.

所述的烷基、烷氧基、卤代烷基、卤代烷氧基、羟烷基、环烷基、杂环烷基、杂芳基任选被一个或多个R ^4A取代； In an optional embodiment, the present disclosure provides a compound represented by formula I, II, III, IV or a pharmaceutically acceptable salt thereof, wherein each of the R ⁴ is independently selected from hydrogen, cyano, halogen, hydroxyl , alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocycloalkyl, cycloalkyloxy, heterocycloalkyloxy, heteroaryl, SO ₂ R', -NHSO ₂ R', COR', -NHCOR', COOR', CONR'(R"), or

The alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocycloalkyl, heteroaryl are optionally substituted by one or more R ^4A ;

Described R ^4A is selected from halogen, cyano, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocycloalkyl, cycloalkyloxy, heterocycloalkyloxy base, SO ₂ R', -NHSO ₂ R', COR', -NHCOR', COOR', CONR'(R") or

In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, IV or pharmaceutically acceptable salts thereof, wherein each R ⁴ is independently selected from hydrogen, cyano or halogen.

In an optional embodiment, the present disclosure provides a compound represented by formula I, II, III, IV or a pharmaceutically acceptable salt thereof, wherein each of the R ⁴ is independently selected from hydrogen, COR', -NHSO ₂ R', SO ₂ R' or

In an optional embodiment, the present disclosure provides a compound represented by formula I, II, III, IV or a pharmaceutically acceptable salt thereof, wherein each of said R ⁴ is independently selected from hydrogen, alkyl, hydroxyalkyl , the alkyl is optionally substituted by one or more R ^4A , the R ^4A is selected from halogen, cyano, alkoxy, SO ₂ R' or

In an optional embodiment, the present disclosure provides a compound represented by formula I, II, III, IV or a pharmaceutically acceptable salt thereof, wherein each of the R ⁴ is independently selected from hydrogen, alkoxy, heterocyclic Alkyloxy, or each R ⁴ is independently selected from C _1-6 alkoxy or 3 to 7 membered heterocycloalkyloxy.

In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, IV or pharmaceutically acceptable salts thereof, wherein said R ¹ is hydrogen.

In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, IV or pharmaceutically acceptable salts thereof, wherein said R ² is selected from isopropyl or methylcyclopropyl.

In an optional embodiment, the present disclosure provides compounds represented by formulas I, II, III, and IV or pharmaceutically acceptable salts thereof, wherein the ring A is pyridyl, and at least one of the R ⁴ is cyano.

In an optional embodiment, the compound shown in the formula I, II, III, IV provided by the present disclosure or a pharmaceutically acceptable salt thereof, wherein, in an optional embodiment, the compound of the formula I, II, III provided in the present disclosure , the compound shown in IV or a pharmaceutically acceptable salt thereof, wherein R' and R" are each independently selected from hydrogen or C _1-6 alkyl.

In an optional embodiment, the compound shown in the formula I, II, III, IV provided by the present disclosure or a pharmaceutically acceptable salt thereof, wherein, in an optional embodiment, the compound of the formula I, II, III provided in the present disclosure , the compound shown in IV or a pharmaceutically acceptable salt thereof, wherein R' and R" are each independently selected from hydrogen or methyl and ethyl.

The compound provided by the present disclosure or a pharmaceutically acceptable salt thereof is selected from:

The present disclosure also provides isotopic substitutions of the compounds described above.

In some embodiments, the isotope substitution is deuterium atom substitution.

The present disclosure also provides a pharmaceutical composition, which comprises the compound represented by the aforementioned formula I, II, III, IV or its pharmaceutically acceptable salt or its isotope substitution and at least one pharmaceutically acceptable carrier and diluent or excipients. In some embodiments, the unit dose of the pharmaceutical composition is 0.001 mg-1000 mg.

In certain embodiments, based on the total weight of the composition, the pharmaceutical composition contains 0.01-99.99% of the compound represented by the aforementioned formula I, II, III, IV or a pharmaceutically acceptable salt thereof or an isotope substitution thing. In certain embodiments, the pharmaceutical composition contains 0.1-99.9% of the compound represented by the aforementioned formulas I, II, III, IV or pharmaceutically acceptable salts or isotope substitutions thereof. In some embodiments, the pharmaceutical composition contains 0.5%-99.5% of the compound represented by the aforementioned formulas I, II, III, IV or pharmaceutically acceptable salts or isotope substitutions thereof. In certain embodiments, the pharmaceutical composition contains 1%-99% of the compound represented by the aforementioned formulas I, II, III, IV or pharmaceutically acceptable salts or isotope substitutions thereof. In certain embodiments, the pharmaceutical composition contains 2%-98% of the compound represented by the aforementioned formulas I, II, III, IV or pharmaceutically acceptable salts or isotope substitutions thereof.

In certain embodiments, the pharmaceutical composition contains 0.01%-99.99% of pharmaceutically acceptable excipients based on the total weight of the composition. In certain embodiments, the pharmaceutical composition contains 0.1%-99.9% of pharmaceutically acceptable excipients. In certain embodiments, the pharmaceutical composition contains 0.5%-99.5% of pharmaceutically acceptable excipients. In certain embodiments, the pharmaceutical composition contains 1%-99% of pharmaceutically acceptable excipients. In certain embodiments, the pharmaceutical composition contains 2%-98% of pharmaceutically acceptable excipients.

The present disclosure also provides a method for preventing and/or treating a patient suffering from a protein-dependent kinase-related disease, by administering to the patient a therapeutically effective amount of the formulas I, II, III, and IV described in the present disclosure. The compound shown or its pharmaceutically acceptable salt or its isotope substitution or the aforementioned pharmaceutical composition.

The present disclosure also provides a method for preventing and/or treating a patient suffering from a cyclin-related disease, by administering to the patient a therapeutically effective amount of the compounds represented by formulas I, II, III, and IV described in the present disclosure. The compound or its pharmaceutically acceptable salt or its isotopic substitution or the aforementioned pharmaceutical composition.

In some embodiments, the protein-dependent kinase-related disease or cyclin-related disease is selected from cell proliferative disease, cancer or immune disease.

In some embodiments, the protein-dependent kinase-associated disease or cyclin-associated disease is selected from breast cancer, ovarian cancer, prostate cancer, melanoma, brain tumor, esophageal cancer, gastric cancer, liver cancer, pancreatic cancer, Colorectal cancer, lung cancer, kidney cancer, skin cancer, glioblastoma, neuroblastoma, sarcoma.

In some specific embodiments, the cancer is selected from cancers in which cyclin E1 and/or cyclin E2 are amplified.

The present disclosure also provides a method for preventing and/or treating a patient suffering from cancer, by administering to the patient a therapeutically effective amount of the compound represented by the formulas I, II, III, IV described in the present disclosure or its alternative Pharmaceutically acceptable salt or its isotope substitution or the aforementioned pharmaceutical composition, the cancer is selected from breast cancer, ovarian cancer, prostate cancer, melanoma, brain tumor, esophageal cancer, gastric cancer, liver cancer, pancreatic cancer, colorectal cancer, Lung cancer, kidney cancer, skin cancer, glioblastoma, neuroblastoma, sarcoma.

The present disclosure provides a therapeutically effective dose of the compounds shown in formulas I, II, III, and IV described in the present disclosure or their pharmaceutically acceptable salts or isotope substitutions or the aforementioned pharmaceutical compositions in the preparation for preventing and/or Use in medicines for the treatment of diseases associated with protein-dependent kinases,

In some specific embodiments, the protein-dependent kinase is selected from CDK2, and the disease related to the protein-dependent kinase is selected from cell proliferative diseases, cancer or immune diseases.

On the other hand, the present disclosure provides a therapeutically effective amount of the compound shown in formula I, II, III, IV described in the present disclosure or its pharmaceutically acceptable salt or its isotope substitution or the aforementioned pharmaceutical composition in preparation for use in Use in medicines for preventing and/or treating diseases related to cell cyclins.

In some specific embodiments, the cyclin is selected from cyclin E, such as cyclin E1, cyclin E2. The cyclin-associated disease is selected from cell proliferative disease, cancer or immune disease.

In some embodiments, the present disclosure provides a therapeutically effective amount of a compound represented by formula I, II, III, IV of the present disclosure or a pharmaceutically acceptable salt thereof or an isotope substitution thereof or the aforementioned pharmaceutical composition in the preparation of a drug for the treatment of cancer Uses in medicine.

In some specific embodiments, the cancer is selected from breast cancer, ovarian cancer, prostate cancer, melanoma, brain tumor, esophageal cancer, gastric cancer, liver cancer, pancreatic cancer, colorectal cancer, lung cancer, kidney cancer, skin cancer Carcinoma, glioblastoma, neuroblastoma, sarcoma.

In some specific embodiments, the cancer is selected from cancers in which cyclin E1 and/or cyclin E2 are amplified.

Another aspect of the present disclosure provides a preparation method of the compound represented by formula III or its pharmaceutically acceptable salts and isotope substitutions, which includes the step of removing the amino protecting group PG from the compound represented by formula V under acidic conditions,

Among them, R ¹ , R ² , R ⁴ , and n are respectively as defined in the compound shown in formula III, and PG is an amino protecting group, which can be tert-butyl, acetyl, trifluoroacetyl, trityl, benzyl base, formyl.

Another aspect of the present disclosure provides a compound represented by formula V,

Among them, R ¹ , R ² , R ⁴ , and n are respectively as defined in the compound shown in formula III, and PG is an amino protecting group, which can be tert-butyl, acetyl, trifluoroacetyl, trityl, benzyl base, formyl.

The pharmaceutically acceptable salts of the compounds described in the present disclosure are selected from inorganic salts or organic salts, and the compounds described in the present disclosure can react with acidic or basic substances to form corresponding salts.

The disclosed compound has a good inhibitory effect on cell cycle protein-dependent kinase, and the IC ₅₀ value of the inhibitory activity to CDK2/Cyclin E is 0.01 to 1000nM, and the IC ₅₀ value of the inhibitory activity of some compounds on CDK2/Cyclin E In the range of 0.01 to 500nM, the IC ₅₀ values of the inhibitory activity of some compounds on CDK2/Cyclin E are in the range of 0.01 to 300nM, and the IC ₅₀ values of the inhibitory activity of some compounds on CDK2/Cyclin E are in the range of 0.01 to 200nM. The _IC50 value of the inhibitory activity of CDK2/Cyclin E is between 0.01 and 100nM, and the IC50 value of the inhibitory activity of some compounds against CDK2/Cyclin E is <100nM.

Some embodiments provide compounds that achieve <50 nM inhibitory activity against CDK2/Cyclin E. Other embodiments provide compounds that achieve <20 nM inhibitory activity against CDK2/Cyclin E. Other embodiments provide compounds that have <10 nM inhibitory activity against CDK2/Cyclin E. Other embodiments provide compounds that achieve <5nM inhibitory activity on CDK2/Cyclin E. Other embodiments provide compounds that achieve <1 nM inhibitory activity against CDK2/Cyclin E.

On the other hand, the disclosed compound also has a good inhibitory effect on CDK2/Cyclin A. Some embodiments provide compounds that achieve <500 nM inhibitory activity against CDK2/Cyclin A. Other embodiments provide compounds that achieve <200 nM inhibitory activity against CDK2/Cyclin A. Other embodiments provide compounds that achieve <150 nM inhibitory activity against CDK2/Cyclin A. Other embodiments provide compounds that achieve <100 nM inhibitory activity against CDK2/Cyclin A. Other embodiments provide compounds that have CDK2/Cyclin A inhibitory activity <50 nM. Other embodiments provide compounds that achieve <20 nM inhibitory activity against CDK2/Cyclin A. Other embodiments provide compounds that achieve <10 nM inhibitory activity against CDK2/Cyclin A.

In other embodiments, the compounds of the present disclosure have weak inhibitory activity on other kinases such as CDK1, CDK4, CDK5, CDK7, CDK9 or GSK3β, and have CDK2-specific selection.

On the other hand, the disclosed compounds may exist in particular geometric or stereoisomeric forms. This disclosure contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers isomers, (D)-isomers, (L)-isomers, and their racemic and other mixtures, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of this disclosure. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers, as well as mixtures thereof, are included within the scope of this disclosure.

In addition, 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 migration of a proton, such as keto-enol and imine-enamine, lactam-lactam, pyrazole base isomerization.

An example of a pyrazolyl equilibrium is between E and F as shown below:

Specific to this application:

All tautomeric forms are within the scope of the invention. The naming of compounds does not exclude any tautomers.

Compounds of the present disclosure may be asymmetric, eg, possess one or more stereoisomers. Unless otherwise stated, all stereoisomers are included, such as enantiomers and diastereomers. Compounds of the present disclosure containing asymmetric carbon atoms can be isolated in optically pure or racemic forms. Optically pure forms can be resolved from racemic mixtures or synthesized by using chiral starting materials or reagents.

Optically active (R)- and (S)-isomers as well as D and L-isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present disclosure is desired, it can be prepared by asymmetric synthesis or derivatization with chiral auxiliary agents, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide 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 diastereoisomeric salt is formed with an appropriate optically active acid or base, and then a diastereomeric salt is formed by a conventional method known in the art. Diastereomeric resolution is performed and the pure enantiomers are recovered. Furthermore, the separation of enantiomers and diastereomers is usually accomplished by the use of chromatography using chiral stationary phases, optionally in combination with chemical derivatization methods (e.g. amines to amino groups formate).

The present disclosure also includes certain isotopically labeled compounds of the disclosure that are identical to those described herein, but wherein one or more atoms are replaced by an atom of an atomic mass or mass number different from that normally found in nature. Examples of isotopes that can be incorporated into 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.

Unless otherwise stated, when a position is specifically designated as deuterium (D), the position is understood to have an abundance of deuterium (i.e., at least 10 % deuterium incorporation). Exemplary compounds having a natural abundance greater than deuterium can be at least 1000 times more abundant deuterium, at least 2000 times more abundant deuterium, at least 3000 times more abundant deuterium, at least 4000 times more abundant deuterium, at least 5000 times more abundant deuterium, at least 6000 times more abundant deuterium, or more abundant deuterium. The present disclosure also includes various deuterated forms of compounds of formula I. Each available hydrogen atom attached to a carbon atom can be independently replaced by a deuterium atom. Those skilled in the art can refer to the relevant literature to synthesize the deuterated form of the compound of formula I. Commercially available deuterated starting materials can be used in the preparation of deuterated forms of compounds of formula I, or they can be synthesized using conventional techniques using deuterated reagents, including but not limited to deuterated borane, trideuterated borane Tetrahydrofuran solution, deuterated lithium aluminum hydride, deuterated ethyl iodide and deuterated methyl iodide, etc.

"Optionally" or "optionally" means that the subsequently described event or circumstance can but need not occur, and that the description includes instances where the event or circumstance occurs or does not occur. For example, "C _1-6 alkyl optionally substituted by halogen or cyano" means that halogen or cyano may but not necessarily exist, and this description includes the case where the alkyl is substituted by halogen or cyano and the alkyl is not substituted by halogen And the case of cyano substitution.

表示未指定构型，即如果化学结构中存在手性异构体，键

可以为

或

或者同时包含

和

两种构型。虽然为简便起见将全部上述结构式画成某些异构体形式，但是本发明可以包括所有的异构体，如互变异构体、旋转异构体、几何异构体、非对映异构体、外消旋体和对映异构体。 In the chemical structure of the compound of the present invention, the bond

Indicates unassigned configuration, i.e. if chiral isomers exist in the chemical structure, the bond

can be

or

or both

and

Two configurations. Although all of the above formulas are drawn as certain isomeric forms for simplicity, the present invention may include all isomers such as tautomers, rotamers, geometric isomers, diastereoisomers body, racemate and enantiomer.

并未指定构型，即可以为Z构型或E构型，或者同时包含两种构型。 In the chemical structures of the compounds described in this disclosure, the bond

If the configuration is not specified, it can be Z configuration or E configuration, or both configurations.

Terminology Explanation

"Pharmaceutical composition" means a mixture containing one or more compounds described herein, or a physiologically acceptable salt or prodrug thereof, and other chemical components, as well as other components such as physiologically acceptable carriers and excipients. Forming agent. The purpose of the pharmaceutical composition is to promote the administration to the organism, facilitate the absorption of the active ingredient and then exert the biological activity.

"Pharmaceutically acceptable excipients" include, but are not limited to, any adjuvants, carriers, glidants, sweeteners, diluents that have been approved by the U.S. Food and Drug Administration (FDA) to be acceptable for human or livestock use , preservative, dye/colorant, flavor enhancer, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier.

An "effective amount" or "therapeutically effective amount" as used in the present disclosure includes an amount sufficient to ameliorate or prevent a symptom or condition of a medical condition. An effective amount also means an amount sufficient to permit or facilitate diagnosis. Effective amounts for a particular patient or veterinary subject may vary depending on factors such as the condition being treated, the general health of the patient, the method, route and dosage of administration, and the severity of side effects. An effective amount may be the maximum dose or dosing regimen that avoids significant side effects or toxic effects.

"Alkyl" refers to a saturated aliphatic hydrocarbon group, including straight and branched chain groups of 1 to 20 carbon atoms. An alkyl group containing 1 to 6 carbon atoms. 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 and various branched isomers, etc. Alkyl groups may be substituted or unsubstituted, and when substituted, substituents may be substituted at any available point of attachment, preferably one or more of the following groups independently selected from halogen, hydroxy, oxo, cyano, amino, C _1-6 alkyl, C _1-6 alkoxy, 3 to 6 membered cycloalkyl or 3 to 6 membered heterocycloalkyl, the alkyl, alkoxy, cycloalkyl or Heterocycloalkyl is optionally substituted with halo, hydroxy, nitro, cyano or amino.

The term "alkenyl" refers to an alkyl compound containing at least one carbon-carbon double bond in the molecule, wherein alkyl is as defined above. Alkenyl groups may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from the group consisting of alkoxy, halo, haloalkyl, haloalkoxy, hydroxy, hydroxyalkane One or more substituents in radical, cyano, amino, nitro, cycloalkyl, heterocycloalkyl, aryl and heteroaryl.

The term "alkynyl" refers to an alkyl compound containing at least one carbon-carbon triple bond in the molecule, wherein alkyl is as defined above. Alkynyl groups may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkoxy, halo, haloalkyl, haloalkoxy, hydroxy, hydroxyalkane One or more substituents in radical, cyano, amino, nitro, cycloalkyl, heterocycloalkyl, aryl and heteroaryl.

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 containing 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, etc.; multicyclic cycloalkyls include spiro Cycloalkyls of rings, parallel rings and bridged rings. Cycloalkyl groups may be substituted or unsubstituted, and when substituted, the substituents may be substituted at any available point of attachment, preferably one or more of the following groups, independently selected from halogen, hydroxy, oxo , cyano, amino, C _1-6 alkyl, C _1-6 alkoxy, 3 to 6 membered cycloalkyl or 3 to 6 membered heterocycloalkyl, the alkyl, alkoxy, cycloalkyl Or heterocycloalkyl is optionally substituted with halo, hydroxy, nitro, cyano or amino.

The term "heterocycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent comprising 3 to 20 ring atoms, one or more of which are selected from nitrogen, oxygen or A heteroatom of S(O) _m (where m is an integer from 0 to 2), excluding ring portions of -OO-, -OS- or -SS-, the remaining ring atoms being carbon. Preferably it contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably it contains 3 to 7 ring atoms. Non-limiting examples of "heterocycloalkyl" include:

等等。

etc.

The heterocycloalkyl ring may be fused to an aryl or heteroaryl ring, wherein the ring bonded to the parent structure is a heterocycloalkyl, non-limiting examples of which include:

等。

wait.

Heterocycloalkyl groups 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, cyano, amino, C _1-6 alkyl, C _1-6 alkoxy, 3 to 6 membered cycloalkyl or 3 to 6 membered heterocycloalkyl, the alkyl, alkoxy, cycloalkyl or heterocycloalkyl optionally Substituted by halogen, hydroxy, nitro, cyano or amino.

The term "alkoxy" refers to -O-(alkyl), wherein alkyl is as defined above. Non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy. Alkoxy may be optionally substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from halogen, hydroxy, oxo, cyano, amino, _{C -6} alkyl, C _1-6 alkoxy, 3 to 7 membered cycloalkyl or 3 to 7 membered heterocycloalkyl, said alkyl, alkoxy, cycloalkyl or heterocycloalkyl are optionally Halogen, hydroxy, nitro, cyano or amino substituted.

Similarly, "cycloalkyloxy" and "heterocycloalkyloxy" are as defined above for "alkoxy", specifically -O-(cycloalkyl), -O-(heterocycloalkyl).

The term "alkylthio" refers to -S-(alkyl), wherein alkyl is as defined above. Non-limiting examples of alkoxy include: methylthio, ethylthio, propylthio, butylthio. Alkylthio may be optionally substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from C _1-6 alkoxy, 3 to 6 membered cycloalkane group, 3 to 6 membered heterocycloalkyl, 3 to 6 membered cycloalkoxy, 3 to 6 membered heterocycloalkyloxy, C _1-6 alkylthio, 3 to 6 membered cycloalkylthio, 3 to 6 6-membered heterocycloalkylthio, the alkoxy, cycloalkyl, heterocycloalkyl, cycloalkoxy, heterocyclyloxy, alkylthio, cycloalkylthio, heterocycloalkylthio are optionally Halogen, hydroxy, cyano or amino substituted.

Similarly, "cycloalkylthio" and "heterocycloalkylthio" have the same definition as the above-mentioned "alkylthio".

"Monovalent group" means that a compound is "formally" eliminated from a monovalent atom or group. "Subunit" refers to a compound formed by "formally" eliminating two monovalent or one divalent atoms or atomic groups.

The term "alkylene" means what remains of an alkane molecule after removal of 2 hydrogen atoms, including straight and branched chain subgroups of 1 to 20 carbon atoms. An alkylene group having 1 to 6 carbon atoms, non-limiting examples include methylene (-CH ₂ -), ethylene (eg -CH ₂ CH ₂ - or -CH(CH ₃ )-). Unless otherwise specified, the alkylene group may be substituted or unsubstituted, and when substituted, the substituent may be substituted at any available point of attachment, preferably one or more of the following groups, independently selected from halogen , hydroxyl, cyano, amino, C _1-6 alkyl or C _1-6 alkoxy.

Similarly, the definitions of "alkyleneoxy", "alkenylene", "alkenyleneoxy", "cycloalkylene", and "heterocycloalkylene" are the same as "alkylene".

The term "aryl" refers to a 6 to 14 membered all-carbon monocyclic or fused polycyclic (that is, rings sharing adjacent pairs of carbon atoms) group, preferably 6 to 12 membered, having a conjugated π-electron system, such as benzene base and naphthyl. The aryl ring may be fused to a heteroaryl, heterocycloalkyl or cycloalkyl ring, where the ring bonded to the parent structure is an aryl ring, non-limiting examples of which include:

Aryl groups may be 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, _C1-6 Alkyl, C _1-6 alkoxy, C _2-6 alkenyloxy, C _2-6 alkynyloxy, 3 to 6-membered cycloalkoxy, 3 to 6-membered heterocycloalkyloxy, C _{3- 8} cycloalkenyloxy, 5 to 6 membered aryl or heteroaryl, said C _1-6 alkyl, C _1-6 alkoxy, C _2-6 alkenyloxy, C _2-6 alkynyloxy, 3 to 6-membered cycloalkoxy, 3 to 6-membered heterocycloalkyloxy, 3 to 8-membered cycloalkenyloxy, 5 to 6-membered aryl or heteroaryl are optionally replaced by one or more members selected from halogen, Hydroxy, cyano, amino, C _1-6 alkyl or C _1-6 alkoxy.

等。 The term "heteroaryl" refers to a heteroaromatic system comprising 1 to 4 heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. The heteroaryl group is preferably 6 to 12 membered, more preferably 5 or 6 membered. For example. Non-limiting examples thereof include: imidazolyl, furyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, isoxazolyl, pyrrolyl, tetrazolyl, pyridyl, pyrimidinyl , Thiadiazole, pyrazinyl, triazolyl, indazolyl, benzimidazolyl,

wait.

The heteroaryl ring may be fused to an aryl, heterocycloalkyl or cycloalkyl ring, wherein the ring bonded to the parent structure is a heteroaryl ring, non-limiting examples of which include:

Heteroaryl may be optionally substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from halogen, hydroxy, cyano, amino, C _alkane group or C _1-6 alkoxy group (here needs to be adjusted according to the claims!).

The term "spiro" refers to a compound in which two rings share one atom. Non-limiting examples of spirocycloalkyl groups include:

The term "merged ring" refers to a compound formed by combining two or more rings by sharing two adjacent atoms. Non-limiting examples of cycloalkyl groups include:

The term "bridged ring" refers to a structure formed by two or more ring structures sharing two non-adjacent ring atoms with each other. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl groups, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of bridged cycloalkyl groups include:

The term "heterocyclic ring" refers to atoms other than carbon atoms that make up the ring, and includes heterocycloalkyl and heteroaryl rings.

The term "hydroxyl" refers to a -OH group.

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

The term "cyano" refers to -CN.

The term "amino" refers to _-NH2 .

The term "nitro" refers to _-NO2 .

The term "oxo" refers to a =O substituent.

"Substituted" means that one or more hydrogen atoms in a group, preferably up to 5, more preferably 1 to 3 hydrogen atoms are independently substituted by the corresponding number of substituents. When a substituent is keto or oxo (ie, =0), then two (2) hydrogens on the atom are replaced.

"Substituted by one or more" means that it can be substituted by single or multiple substituents. When substituted by a plurality of substituents, it may be a plurality of the same substituents, or one or a combination of a plurality of different substituents.

Detailed ways

The present disclosure is further described below in conjunction with examples, but these examples do not limit the scope of the present disclosure.

The experimental methods in the embodiments of the present disclosure that do not indicate specific conditions are generally in accordance with conventional conditions, or in accordance with the conditions suggested by the raw material or commodity manufacturers. Reagents without specific sources indicated are conventional reagents purchased in the market.

Compound structures were determined by nuclear magnetic resonance (NMR) or/and mass spectroscopy (MS). NMR shifts (δ) are given in units of 10 ^-6 (ppm). The determination of NMR is to use Bruker AVANCE-400 nuclear magnetic instrument, and the determination solvent is deuterated dimethyl sulfoxide (DMSO-d ₆ ), deuterated chloroform (CDCl ₃ ), deuterated methanol (CD ₃ OD), and the internal standard is four Methylsilane (TMS).

MS was determined by Shimadzu 2010 Mass Spectrometer or Agilent 6110A MSD mass spectrometer.

The determination of HPLC uses Shimadzu LC-20A systems, Shimadzu LC-2010HT series or Agilent Agilent 1200LC high pressure liquid chromatography (Ultimate XB-C18 3.0*150mm column or Xtimate C18 2.1*30mm column).

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 were used for chiral HPLC analysis and determination 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 column;

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

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

The chiral preparative column uses DAICEL CHIRALPAK IC (250mm*30mm, 10um) or Phenomenex-Amylose-1 (250mm*30mm, 5um).

The CombiFlash rapid preparation instrument uses Combiflash Rf150 (TELEDYNE ISCO).

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 ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Shaoyuan Chemical Technology (Accela ChemBio Inc), Darui chemical companies.

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

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

The hydrogen atmosphere means that the reaction bottle is connected to a hydrogen balloon with a capacity of about 1L.

The pressurized hydrogenation reaction uses Parr 3916EKX hydrogenation instrument and Qinglan QL-500 hydrogen generator or HC2-SS hydrogenation instrument.

The hydrogenation reaction is usually vacuumized and filled with hydrogen, and the operation is repeated 3 times.

For the microwave reaction, a CEM Discover-S 908860 microwave reactor was used.

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

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

The monitoring of the reaction process in the embodiment adopts thin-layer chromatography (TLC), the developing agent used in reaction, the eluent system of the eluent system of the column chromatography that purification compound adopts and the developing agent system of thin-layer chromatography, the volume of solvent The ratio is adjusted according to the polarity of the compound, and it can also be adjusted by adding a small amount of basic or acidic reagents such as triethylamine and acetic acid.

Example 1

(1R,3S)-3-(3-((4-cyanopyridin-2-yl)amino)-1H-pyrazol-5-yl)cyclopentylisopropylcarbamate

first step

(1R,3S)-3-(1-(tert-butyl)-5-((4-cyanopyridin-2-yl)amino)-1H-pyrazol-3-yl)cyclopentylisopropyl carbamate 1b

(1R,3S)-3-(5-Amino-1-(tert-butyl)-1H-pyrazol-3-yl)cyclopentyl isopropyl carbamate (40mg, 0.13mmol, using patent Prepared by the method disclosed in the application "WO 2020/157652 A2"), 2-chloroisonicotinonitrile (20mg, 0.14mmol), palladium acetate (5.8mg, 0.03mmol), 1,1'-binaphthalene-2.2' - Diphenylphosphine (32.3 mg, 0.05 mmol), cesium carbonate (84.5 mg, 0.26 mmol) and 1,4-dioxane (1.5 mL) were placed in a microwave tube and blown with nitrogen for 1 minute. The reaction solution was microwaved at 100° C. for 2 hours. After the reaction, it was cooled to room temperature and concentrated under reduced pressure to obtain the crude product 1b as a black solid, which was directly used in the next reaction (80 mg, yield 30%).

MS m/z (ESI): 411.4 [M+H] ⁺ .

second step

(1R,3S)-3-(3-((4-cyanopyridin-2-yl)amino)-1H-pyrazol-5-yl)cyclopentylisopropylcarbamate 1

At room temperature, to (1R,3S)-3-(1-(tert-butyl)-5-((4-cyanopyridin-2-yl)amino)-1H-pyrazol-3-yl) ring Formic acid (1.5 mL) was added to pentyl isopropyl carbamate (80 mg, 0.04 mmol). The reaction solution was stirred and reacted at 100° C. for 1 hour. After the reaction, the reaction solution was directly purified by C-18 reverse phase chromatography to obtain the title compound 1 (5.4 mg, yield 38%).

MS m/z (ESI): 355.4 [M+H] ⁺ .

¹ H NMR(400MHz,DMSO):δ11.99(s,1H),9.69(s,1H),8.33(s,1H),7.67(s,1H),7.00(d,J＝35.1Hz,2H) ,6.08(s,1H),5.00(s,1H),3.58(s,1H),3.06(s,1H),1.96(m,3H),1.67(m,3H),1.13-0.92(m,6H ).

Example 2

(1R,3S)-3-(3-((5-(cyanomethyl)pyridin-2-yl)amino)-1H-pyrazol-5-yl)cyclopentylisopropylcarbamate

first step

(1R,3S)-3-(1-(tert-butyl)-5-((5-(cyanomethyl)pyridin-2-yl)amino)-1H-pyrazol-3-yl)cyclopentene isopropyl carbamate 2b

Under nitrogen atmosphere, 2-(6-chloropyridin-3-yl)acetonitrile (20mg, 0.13mmol), (1R,3S)-3-(5-amino-1-(tert-butyl)-1H -pyrazol-3-yl) cyclopentyl isopropyl carbamate (40.4mg, 0.13mmol, prepared by the method disclosed in the patent application "WO 2020/157652 A2"), palladium acetate (5.9mg, 0.03 mmol), 1,1'-binaphthyl-2,2'-bisdiphenylphosphine (16.3 mg, 0.03 mmol) were dissolved in 1,4-dioxane (3 mL). Add cesium carbonate (106.8mg, 0.33mmol) and react at 80°C for 2 hours. The reaction solution was cooled to room temperature, filtered, and the filtrate was collected and concentrated under reduced pressure to obtain the title compound 2b (40 mg, yield 71.8%).

MS m/z (ESI): 425.6 [M+H] ⁺ .

second step

(1R,3S)-3-(3-((5-(cyanomethyl)pyridin-2-yl)amino)-1H-pyrazol-5-yl)cyclopentylisopropylcarbamate 2

(1R,3S)-3-(1-(tert-butyl)-5-((5-(cyanomethyl)pyridin-2-yl)amino)-1H-pyrazol-3-yl)ring Amyl isopropyl carbamate (40mg, 0.09mmol) was dissolved in 6mL of formic acid and reacted at 80°C for 3 hours. After cooling to room temperature, the reaction solution was directly purified by C-18 reverse phase chromatography to obtain the title compound 2 (12 mg, yield 34.5%).

MS m/z (ESI): 369.4 [M+H] ⁺ .

¹ H NMR(400MHz,DMSO):δ11.84(s,1H),9.20(s,1H),8.06(d,J=2.4Hz,1H),7.52(dd,J=8.7,2.5Hz,1H) ,7.29(s,1H),6.96(d,J=7.9Hz,1H),6.07(s,1H),5.00(s,1H),3.88(s,2H),3.58(m,2H),3.11– 2.97(m,1H),2.00(t,J=8.3Hz,1H),1.90(m,1H),1.73(d,J=9.8Hz,2H),1.61(s,1H),1.03(d,J = 6.5, 6H).

Example 3

(1R,3S)-3-(3-((4-(1H-1,2,4-triazol-1-yl)pyridin-2-yl)amino)-1H-pyrazol-5-yl)ring Amyl isopropyl carbamate

first step

2-Chloro-4-(1H-1,2,4-triazol-1-yl)pyridine 3b

1H-1,2,4-triazole (420 mg, 6.08 mmol) was dissolved in N,N-dimethylformamide (8 mL) under nitrogen atmosphere. Sodium hydride (243.2 mg, 6.08 mmol) was added to the reaction solution in batches at room temperature, stirred at room temperature for half an hour, 2,4-dichloropyridine (300 mg, 2.02 mmol) was added to the reaction solution in batches, Stir at 100°C for 2 hours, cool down to room temperature, add 50ml of water to the reaction solution, extract with ethyl acetate, combine the organic phases, wash the organic phases, dry, and concentrate under reduced pressure, the residue is purified by C-18 reverse phase chromatography The title compound 3b (120 mg, 32.8% yield) was obtained.

MS m/z (ESI): 181.2 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO): δ9.58(s, 1H), 8.58(d, J=5.5Hz, 1H), 8.38(s, 1H), 8.11(d, J=2.0Hz, 1H), 7.95 (dd, J = 5.6, 1.9 Hz, 1H).

second step

(1R,3S)-3-(5-((4-(1H-1,2,4-triazol-1-yl)pyridin-2-yl)amino)-1-(tert-butyl)-1H -pyrazol-3-yl)cyclopentylisopropylcarbamate 3c

Under nitrogen atmosphere, 2-chloro-4-(1H-1,2,4-triazol-1-yl)pyridine (15 mg, 0.08 mmol), (1R,3S)-3-(5-amino-1 -(tert-butyl)-1H-pyrazol-3-yl)cyclopentylisopropylcarbamate (25.6mg, 0.08mmol, prepared by the method disclosed in the patent application "WO 2020/157652 A2" ), palladium acetate (3.7mg, 0.02mmol), 1,1'-binaphthyl-2,2'-bisdiphenylphosphine (10.3mg, 0.02mmol) were dissolved in 1,4-dioxane (2mL) . Add cesium carbonate (67.7mg, 0.21mmol) and react at 80°C for 2 hours. The reaction solution was cooled to room temperature, filtered, and the filtrate was collected and concentrated under reduced pressure to obtain the title compound 3c (30 mg, yield 79.8%)

MS m/z (ESI): 453.7 [M+H] ⁺ .

third step

(1R,3S)-3-(3-((4-(1H-1,2,4-triazol-1-yl)pyridin-2-yl)amino)-1H-pyrazol-5-yl)ring Amyl isopropyl carbamate 3

(1R,3S)-3-(5-((4-(1H-1,2,4-triazol-1-yl)pyridin-2-yl)amino)-1-(tert-butyl)- 1H-pyrazol-3-yl) cyclopentyl isopropyl carbamate (30mg, 0.07mmol) was dissolved in 3mL formic acid, and reacted at 80°C for 12 hours. After cooling to room temperature, the reaction solution was directly purified by C-18 reverse phase chromatography to obtain the title compound 3 (10.7 mg, yield 40.7%).

MS m/z (ESI): 397.4 [M+H] ⁺ .

¹ H NMR(400MHz,DMSO-d6):δ12.21-11.66(m,1H),9.52-9.42(m,1H),9.42-9.33(m,1H),8.34-8.18(m,2H),7.81 (s,1H),7.20(d,J＝2.1Hz,1H),7.04-6.83(m,1H),6.05(s,1H),5.05-4.95(m,1H),3.69-3.47(m,3H ),2.10-1.96(m,1H),1.95-1.83(m,1H),1.79-1.67(m,2H),1.66-1.54(m,1H),1.03(br d,J＝6.4Hz,6H) .

Example 4

(Propan-2-ylamino)methane-(1R,3S)-3-[5-({4-[(methylamino)dioxylidene-6-thio]phenyl}amino)-2H- Pyrazol-3-yl]cyclopentyl ester

Compound 4 was prepared according to the synthesis steps of Example 1 by replacing the corresponding raw materials.

MS m/z (ESI): 422.2 [M+H] ⁺ .

Example 5

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-(5-{[5-(cyanomethyl)pyrazin-2-yl]amino}-2H-pyrazol-3-yl ) cyclopentyl ester

first step

2-(5-Bromopyrazin-2-yl)-2-cyanoacetic acid-2-methylpropan-2-yl ester 5b

2-Bromo-5-chloropyrazine 5a (500mg, 2.6mmol) was first dissolved in N,N-dimethylformamide (20ml) followed by the addition of tert-butyl 2-cyanoacetate (733mg, 5.2mmol) , cesium carbonate (2.1g, 6.5mmol), react at room temperature for 4 hours. The crude product is obtained after extraction, drying and concentration, which can be directly used in the next reaction.

MS m/z (ESI): 298.4 [M+H] ⁺ .

second step

(5-Bromopyrazin-2-yl)acetonitrile 5c

2-(5-Bromopyrazin-2-yl)-2-cyanoacetic acid-2-methylpropan-2-yl ester 5b was dissolved in dichloromethane (10 ml), followed by the addition of excess trifluoroacetic acid ( 2ml), react at room temperature for 1 hour. The crude product was obtained after extraction, drying and concentration, and (5-bromopyrazin-2-yl)acetonitrile 5c (150 mg, 29%) was obtained by normal phase column chromatography.

MS m/z (ESI): 198.2 [M+H] ⁺ .

third step

(Propan-2-ylamino)methane acid-(1R,3S)-3-(5-{[5-(cyanomethyl)pyrazin-2-yl]amino}-1-(2-methylpropane -2-yl)pyrazol-3-yl)cyclopentyl ester 5d

Under nitrogen atmosphere, (5-bromopyrazin-2-yl)acetonitrile (50mg, 0.25mmol), (1R,3S)-3-(5-amino-1-(tert-butyl)-1H-pyridine Azol-3-yl) cyclopentyl isopropyl carbamate (77mg, 0.25mmol, prepared by the method disclosed in the patent application "WO 2020/157652 A2"), XantPhos (8.7mg 0.015mmol), Pd2dba3 ( 11.4mg, 0.0125mmol), cesium carbonate (81.2mg, 0.25mmol) were dissolved in dioxane (2mL), and microwaved at 120°C for 1 hour. The reaction solution was cooled to room temperature, filtered, and the filtrate was collected, concentrated under reduced pressure and separated from the normal phase to obtain the title compound 5d (50 mg, yield 47%).

MS m/z (ESI): 426.2 [M+H] ⁺ .

the fourth step

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-(5-{[5-(cyanomethyl)pyrazin-2-yl]amino}-2H-pyrazol-3-yl ) cyclopentyl ester 5

(Propan-2-ylamino)methane acid-(1R,3S)-3-(5-{[5-(cyanomethyl)pyrazin-2-yl]amino}-1-(2-methyl Propan-2-yl)pyrazol-3-yl)cyclopentyl ester (40mg, 0.09mmol) was dissolved in 3mL formic acid and reacted at 100°C for 1 hour. After cooling to room temperature, the reaction solution was purified by C-18 reverse phase chromatography to obtain the title compound 5 (8 mg, yield 23.0%).

MS m/z (ESI): 370.2 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO): δ12.00(s, 1H), 9.70(s, 1H), 8.55(s, 1H), 8.11(d, J=1.4Hz, 1H), 6.94(d, J= 7.7Hz, 1H), 6.19(s, 1H), 5.00(s, 1H), 4.05(s, 2H), 3.63–3.54(m, 1H), 3.11-3.02(m, 1H), 2.10-1.84(m , 3H), 1.79-1.68 (m, 2H), 1.64-1.60 (m, 1H), 1.04 (dJ=6.7Hz, 6H).

Example 6

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-{5-[(5-cyanopyrazin-2-yl)amino]-2H-pyrazol-3-yl}cyclopentyl ester

Compound 6 was prepared according to the similar steps of Example 1 by substituting corresponding raw materials and reagents.

MS m/z (ESI): 356.4 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ): δ12.21(s, 1H), 10.47(s, 1H), 7.93(d, J=9.6Hz, 1H), 7.69(d, J=Hz, 1H) ,6.93(d,J=7.2Hz,1H),6.19(s,1H),5.01(d,J=4.4Hz,1H),3.62-3.54(m,1H),3.11-3.05(m,1H), 2.08-1.58 (m, 6H), 1.03 (d, J=6.4Hz, 6H).

Example 7

(Propan-2-ylamino)methane-(1R,3S)-3-{5-[(6-cyano-1,2-diazacyclohexyl-3-yl)amino]-2H-pyridine Azol-3-yl}cyclopentyl ester

Compound 7 was prepared according to the synthesis steps of Example 1 by replacing the corresponding raw materials.

MS m/z (ESI): 356.4 [M+H] ⁺ .

Example 8

(Propan-2-ylamino)methane-(1R,3S)-3-[5-({4-[(amino)dioxylidene-6-thio]phenyl}amino)-2H-pyrazole -3-yl]cyclopentyl ester

first step

(1R,3S)-3-(1-(tert-butyl)-5-((4-sulfonylphenyl)amino)-1H-pyrazol-3-yl)cyclopentylisopropylcarbamate 8b

Under nitrogen atmosphere, 4-bromobenzenesulfonamide (23.5mg, 0.10mmol), (1R,3S)-3-(5-amino-1-(tert-butyl)-1H-pyrazol-3-yl ) cyclopentyl isopropyl carbamate (30.8mg, 0.10mmol, prepared by the method disclosed in the patent application "WO 2020/157652 A2"), XantPhos Pd G3 (9.5mg, 0.01mmol), potassium phosphate ( 63.6mg, 0.30mmol) was dissolved in NMP (2mL), and reacted under microwave at 80°C for 2 hours. The reaction solution was cooled to room temperature, filtered, and the filtrate was collected, concentrated under reduced pressure and separated from the normal phase to obtain the title compound 8b (40 mg, yield 85%).

MS m/z (ESI): 464.3 [M+H] ^+.

third step

(1R,3S)-3-(3-((4-sulfonylphenyl)amino)-1H-pyrazol-5-yl)cyclopentylisopropylcarbamate 8

8b (40mg, 0.08mmol) was dissolved in 3mL formic acid and reacted at 80°C for 12 hours. After cooling to room temperature, the reaction solution was purified by C-18 reverse phase chromatography to obtain the title compound 8 (7.7 mg, yield 18.9%).

MS m/z (ESI): 408.2 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ): δ11.87(s, 1H), 8.86(s, 1H), 7.64-7.57(m, 2H), 7.40(d, J=8.4Hz, 2H), 7.03 -6.91(m,3H),5.69(s,1H),5.00(s,1H),3.63-3.53(m,1H),3.11-3.01(m,1H),2.49-2.41(m,1H),2.05 -1.99 (m, 1H), 1.97-1.84 (m, 1H), 1.73 (m, 2H), 1.27-1.22 (m, 1H), 1.08-1.00 (d, J=6.6, 6H).

Example 9

(Propan-2-ylamino)methane-(1R,3S)-3-[5-({4-[(amino)dioxylidene-6-sulfanyl]-2fluorophenyl}amino)-2H -pyrazol-3-yl]cyclopentyl ester

first step

(Propan-2-ylamino)methane acid-(1R,3S)-3-(5-{[4-(Aminodioxyylidene-6-thio)-2-fluorophenyl]amino}-1- (2-Methylprop-2-yl)pyrazol-3-yl)cyclopentyl ester 9b

Compound 9a (2.7g, 10.6mmol), 1a (3.0g, 9.7mmol), Xantphos-Pd-G3 (300mg, 0.3mmol) and potassium phosphate (2.5g, 11.6mmol), were dissolved in N-methylpyrrolidone ( 15 mL), react under microwave at 120°C for 5 hours. After the reaction was complete, water (30 mL) was added to the reaction solution, and extracted with EtOAc (20 mL×2). The organic phases were combined and dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated in vacuo to obtain the crude compound. The crude product was purified by C18 column to obtain compound 9b (4.0 g, yield 85.7%).

MS m/z (ESI): 482.2 [M+H] ⁺ .

second step

(Propan-2-ylamino)methane-(1R,3S)-3-[5-({4-[(amino)dioxylidene-6-sulfanyl]-2fluorophenyl}amino)-2H -pyrazol-3-yl]cyclopentyl ester 9

Compound 9b (4.0 g, 8.3 mmol) was dissolved in formic acid (20 mL), and reacted at 120° C. for 5 hours. After the reaction was complete, the reaction solution was concentrated to obtain a crude compound, which was purified by reverse-phase HPLC to obtain compound 9 (1.7 g, yield 48.2%).

MS m/z (ESI): 426.5 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ): δ11.96(s, 1H), 8.62(s, 1H), 8.19(t, J=8.5Hz, 1H), 7.51(d, 2H), 7.17(s ,2H),6.94(d,J=7.8Hz,1H),5.83(s,1H),5.08-4.96(m,1H),3.69-3.48(m,1H),3.16-2.92(m,1H), 2.48-2.40(m,1H),2.09-1.98(m,1H),1.96-1.86(m,1H),1.76-1.69(m,2H),1.66-1.55(m,1H),1.04(d,J = 6.6, 6H).

Example 10

{[(2S)-Butan-2-yl]amino}methane-(1R,3S)-3-(5-{[4-(aminodioxyylidene-6-thio)-2-fluorophenyl ]amino}-2H-pyrazol-3-yl)cyclopentyl ester

first step

{[(2S)-Butan-2-yl]amino}methane-(1R,3S)-3-(5-{[4-(aminodioxyylidene-6-thio)-2-fluorophenyl ] amino }-1-(2-methylprop-2-yl)pyrazol-3-yl)cyclopentyl ester 10c

Compound 10a (0.5g, 1.96mmol), 10b (0.65g, 2.0mmol, prepared by the method disclosed in the patent application "WO 2020/157652 A2"), Xantphos-Pd-G3 (95mg, 0.1mmol), phosphoric acid Potassium (0.53g, 2.5mmol) was dissolved in N-methylpyrrolidone (10mL), and reacted under microwave at 120°C for 5 hours. After the reaction was complete, water (30 mL) was added to the reaction liquid, and extracted with EtOAc (20 mL×2). The organic phases were combined and dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated in vacuo to obtain the crude compound. The crude product was purified by C18 silica gel column to obtain compound 10c (454 mg, 0.91 mmol, yield: 46.4%).

MS(ESI)m/z 496.4[M+H] ⁺

second step

{[(2S)-Butan-2-yl]amino}methane-(1R,3S)-3-(5-{[4-(aminodioxyylidene-6-thio)-2-fluorophenyl ]amino}-2H-pyrazol-3-yl)cyclopentyl ester 10

Compound 10c (454mg, 0.91mmol) was dissolved in formic acid (10mL) and reacted at 120°C for 5 hours. After the reaction was complete, the reaction solution was concentrated to obtain a crude compound, which was purified by HPLC to obtain compound 10 (250 mg, 0.57 mmol, yield: 62.5%).

MS m/z (ESI): 440.2 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ): δ11.95(s, 1H), 8.61(s, 1H), 8.19(t, J=8.5Hz, 1H), 7.58-7.46(m, 2H), 7.17 (s,2H),6.88(d,J＝8.3Hz,1H),5.83(s,1H),5.07-4.95(m,1H),3.47-3.35(m,1H),3.15-2.99(m,1H ),2.49-2.40(m,1H),2.11-1.99(m,1H),1.98-1.84(m,1H),1.80-1.66(m,2H),1.66-1.53(m,1H),1.43-1.31 (m, 2H), 1.02 (d, J=6.6Hz, 3H), 0.81 (t, J=7.4Hz, 3H).

Example 11

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-[5-({6-[(methylamino)dioxylidene-6-thio]pyridin-3-yl}amino) -2H-pyrazol-3-yl]cyclopentyl ester

first step

5-Bromo-N-methylpyridine-2-sulfonamide 11b

At room temperature, dissolve methylamine hydrochloride (180mg, 3mmol) and DIEA (650mg, 5mmol) in anhydrous dichloromethane (10mL), and slowly add 5-bromopyridine-2-sulfonyl chloride (250mg, 1mmol), continue to stir, TLC monitoring until the end of the reaction, add water (10mL) to wash once, saturated brine (10mL) wash once, collect the organic phase layer anhydrous sodium sulfate and dry, spin to a small volume direct normal phase column Chromatography afforded 11b (180 mg, 72% yield).

second step

(1R,3S)-3-(1-(tert-butyl)-5-((6-(N-methylaminosulfonyl)pyridin-3-yl)amino)-1H-pyrazol-3-yl) Cyclopentyl isopropyl carbamate 11c

Under nitrogen atmosphere, 5-bromo-N-methylpyridine-2-sulfonamide (25.1 mg, 0.10 mmol), (1R,3S)-3-(5-amino-1-(tert-butyl)- 1H-pyrazol-3-yl) cyclopentyl isopropyl carbamate (30.8 mg, 0.10 mmol, prepared by the method disclosed in the patent application "WO 2020/157652 A2"), Pd ₂ (dba) ₃ (9.1mg, 0.01mmol), Xantphos (11.4mg, 0.02mmol), CS ₂ CO ₃ (96.2mg, 0.30mmol) were dissolved in 1,4-dioxane (2mL), and reacted under microwave at 80°C 2 Hour. The reaction solution was cooled to room temperature, filtered, and the filtrate was collected, concentrated under reduced pressure and separated from the normal phase to obtain the title compound 11c (15 mg, yield 32%).

MS m/z (ESI): 479.3 [M+H] ⁺ .

third step

(1R,3S)-3-(3-((6-(N-Methylsulfonyl)pyridin-3-yl)amino)-1H-pyrazol-5-yl)cyclopentylisopropylaminomethyl Ester 11

((1R,3S)-3-(1-(tert-butyl)-5-((6-(N-methylaminosulfonyl)pyridin-3-yl)amino)-1H-pyrazole-3- Base) cyclopentyl isopropyl carbamate (15 mg, 0.03 mmol) was dissolved in 2 mL of formic acid and reacted at 80 ° C for 12 hours. Cooled to room temperature, the reaction solution was purified by C-18 reverse phase chromatography to obtain the title Compound (2.2 mg, yield 17.3%).

MS m/z (ESI): 423.2 [M+H] ⁺ .

Example 12

{[(2S)-Butan-2-yl]amino}methane-(1R,3S)-3-[5-({6-[(Ethylamino)dioxyylidene-6-thio]pyridine- 3-yl}amino)-2H-pyrazol-3-yl]cyclopentyl ester

Compound 12 was prepared according to the synthetic steps of Example 11 by replacing the corresponding raw materials.

MS m/z (ESI): 451.2 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d6): δ12.00(s, 1H), 9.02(s, 1H), 8.57(d, J=1.6Hz, 1H), 7.97(dd, J ₁ =1.6Hz, J ₂ =8.4Hz,1H),7.73(d,J=8.8Hz,1H),7.43(t,J=5.6Hz,1H),6.87(d,J=8.4Hz,1H),5.72(s,1H) ,5.00(d,J＝2.4Hz,1H),3.42-3.35(m,1H),3.09-3.05(m,1H),2.90-2.83(m,2H),2.08-1.56(m,6H),1.40 -1.30(m, 2H), 1.01(d, J=6.4Hz, 3H), 0.96(t, J=7.2Hz, 3H), 0.80(t, J=7.6Hz, 3H).

Example 13

(1R,3S)-3-(3-((5-(2-(Deuteromethylamino)-2-oxoethyl)pyrazin-2-yl)amino)-1H-pyrazol-5-yl ) cyclopentyl (S)-sec-butyl carbamate

first step

Diethyl 2-(5-chloropyrazin-2-yl)malonate 13b

Under nitrogen atmosphere, diethyl malonate (2.49g, 15.51mmol), 2-pyridinecarboxylic acid (127.3mg, 1.04mmol), cesium carbonate (5.06g, 15.51mmol) and cuprous iodide (393.9mg, 2.07 mmol) was added to a solution of 2-bromo-5-chloropyrazine 13a (1 g, 5.17 mmol) in dioxane (30 mL). Reacted overnight at 105°C, lowered to room temperature, concentrated the reaction solution under reduced pressure, added 30 mL of water, extracted with ethyl acetate, combined the organic phases, washed the organic phase with saturated brine, dried, and concentrated under reduced pressure. Separation and purification by silica gel column chromatography gave the title compound 13b (1.1 g, yield 79%).

MS m/z (ESI): 273.1 [M+H] ⁺ .

second step

2-(5-Chloropyrazin-2-yl) ethyl acetate 13c

Diethyl 2-(5-chloropyrazin-2-yl)malonate 13b (1 g, 3.68 mmol) was dissolved in N,N-dimethylsulfoxide (20 mL), and sodium chloride ( 234.6mg, 4mmol) and water (132.4mg, 7.35mmol). React at 145°C. After the reaction is completed, the reaction solution is lowered to room temperature, 30 mL of water is added, extracted with ethyl acetate, the organic phases are combined, the organic phase is washed with saturated brine, dried, and concentrated under reduced pressure, and the residue is washed with silica gel Purified by column chromatography to obtain the title compound 13c (514.7 mg, yield 70%).

MS m/z (ESI): 201.1 [M+H] ⁺ .

third step

2-(5-((1-(tert-butyl)-3-((1S,3R)-3-((((S)-sec-butyl)carbamoyl)oxy)cyclopentyl)-1H -pyrazol-5-yl)amino)pyrazin-2-yl)ethyl acetate 13d

Ethyl 2-(5-chloropyrazin-2-yl)acetate (500mg, 2.5mmol), (1R,3S)-3-(5-amino-1-(tert-butyl)-1H-pyrazole- 3-yl) cyclopentyl ((S)-sec-butyl) carbamate (805mg, 2.5mmol, prepared by the method disclosed in the patent application "WO 2020/157652 A2"), cesium carbonate (2.45g, 7.5mmol) and XantPhos Pd G3 (119mg, 0.125mmol) were dissolved in 1,4-dioxane (10mL), and refluxed under nitrogen protection. After the reaction was completed, the reaction solution was lowered to room temperature, and 20ml of water was added. The ester was extracted, and the organic phases were combined. The organic phase was washed with saturated brine, dried, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography to obtain the title compound 13d (850.5 mg, yield 70%).

MS m/z (ESI): 487.5 [M+H] ⁺ .

the fourth step

2-(5-((1-(tert-butyl)-3-((1S,3R)-3-((((S)-sec-butyl)carbamoyl)oxy)cyclopentyl)-1H -pyrazol-5-yl)amino)pyrazin-2-yl)acetic acid 13e

2-(5-((1-(tert-butyl)-3-((1S,3R)-3-((((S)-sec-butyl)carbamoyl)oxy)cyclopentyl)- 1H-pyrazol-5-yl)amino)pyrazin-2-yl)ethyl acetate 13d (500mg, 1.03mmol) was dissolved in THF (4mL), and to the solution was added aqueous lithium hydroxide (2M, 2.06mmol) . Reaction at room temperature, after the reaction was completed, the reaction solution was acidified with 1M hydrochloric acid, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried, and concentrated under reduced pressure to obtain the title compound 13e (424.6 mg, yield 90 %).

MS m/z (ESI): 457.4 [MH] ^- .

the fifth step

(1R,3S)-3-(1-(tert-butyl)-5-((5-(2-(deuteromethylamino)-2-oxoethyl)pyrazin-2-yl)amino)- 1H-pyrazol-3-yl)cyclopentyl((S)-sec-butyl)carbamate 13f

13e (400mg, 0.87mmol) was dissolved in dichloromethane (5mL), 1,1'carbonyldiimidazole (141.6mg, 0.88mmol) was added to the solution, and the reaction solution was stirred at room temperature for 1 hour. Deuteromethylamine hydrochloride (62.2 mg, 0.87 mmol) was dissolved in dichloromethane (1 mL), triethylamine (138 μL, 1 mmol) was added thereto, and stirred at room temperature for 30 minutes. The suspension was slowly added to the above mixture, and the reaction was continued overnight. The reaction solution was quenched with water, extracted with dichloromethane, the organic phases were combined, and the organic phase was successively washed with 1M hydrochloric acid, 1M aqueous sodium hydroxide solution and saturated brine, dried, concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography to obtain The title compound 13f (247.4 mg, 60% yield).

MS m/z (ESI): 475.5 [M+H] ⁺ .

step six

(1R,3S)-3-(3-((5-(2-(Deuteromethylamino)-2-oxoethyl)pyrazin-2-yl)amino)-1H-pyrazol-5-yl ) cyclopentyl-(S)-sec-butylcarbamate 13

13f (173mg, 0.36mmol) was dissolved in formic acid (3mL), and the reaction solution was reacted at 100°C for about 2 hours. After the reaction solution was cooled to room temperature and concentrated under reduced pressure, the residue was purified by C-18 reverse phase chromatography to obtain the title compound 13 (30 mg, yield 20%).

MS m/z (ESI): 419.4 [M+H] ⁺ .

Example 14

(1R,3S)-3-(3-((5-(cyanomethyl)-3-methylpyridin-2-yl)amino)-1H-pyrazol-5-yl)isopropylcarbamate cyclopentyl ester

first step

4-(6-Chloro-5-methylpyridin-3-yl)isoxazole 14b

Under nitrogen atmosphere, 5-bromo-2-chloro-3-methylpyridine (1g, 4.84mmol) was dissolved in N,N-dimethylformamide (5mL) and water (5ml), and 4-iso Oxazole boronic acid pinacol ester (1.04g, 5.32mmol), potassium fluoride (0.84g, 14.52mmol), Pd(dppf)Cl ₂ (0.35g, 0.48mmol), rise to 80°C for 16h, then cool to room temperature , The reaction solution was poured into 10ml of water, extracted with ethyl acetate (10ml*2), the organic phases were combined, washed, dried, and concentrated under reduced pressure to obtain the title compound 14b (1g, yield 105%).

MS m/z (ESI): 195.2 [M+H] ⁺ .

second step

2-(6-Chloro-5-methylpyridin-3-yl)acetonitrile 14c

Dissolve 4-(6-chloro-5-methylpyridin-3-yl)isoxazole (1g, 5mmol) in methanol (10ml) and water (1ml), then add potassium fluoride (60mg, 1mmol), Raise the temperature to 90°C, react for 1h, cool down to room temperature, spin off methanol, add 3ml of water, extract with ethyl acetate (5ml*2), combine the organic phases, mix the sample and pass through the column to obtain the title compound 14c (350mg, yield 40.9% ).

MS m/z (ESI): 167.2 [M+H] ⁺ .

third step

(1R,3S)-3-(1-(tert-butyl)-5-((5-(cyanomethyl)-3-methylpyridin-2-yl)amino)-1H-pyrazol-3-yl ) Cyclopentyl isopropylcarbamate 14d

14c (35.8 mg, 0.21 mmol), (1R,3S)-3-(5-amino-1-(tert-butyl)-1H-pyrazol-3-yl)cyclopentylisopropylaminomethyl Ester (60mg, 0.19mmol) prepared by the method disclosed in the patent application "WO 2020/157652 A2"), Pd2(dba)3 (17.8mg, 0.02mmol), 1,1'-binaphthyl-2,2 '-Bisdiphenylphosphine (24.3 mg, 0.04 mmol) was dissolved in 1,4-dioxane (2 mL). Potassium phosphate (103.4mg, 0.49mmol) was added, and microwaved at 120°C for 4h. The reaction solution was cooled to room temperature, poured into 5ml of water, extracted with ethyl acetate (5ml*2), the organic phases were combined, mixed and passed through the column to obtain the title compound 14d (40mg, yield 46.9%).

MS m/z (ESI): 439.3 [M+H] ⁺ .

the fourth step

(1R,3S)-3-(3-((5-(cyanomethyl)-3-methylpyridin-2-yl)amino)-1H-pyrazol-5-yl)isopropylcarbamate cyclopentyl Ester 14

14d (40mg, 0.07mmol) was dissolved in 4mL formic acid and reacted at 100°C for 3 hours. After cooling to room temperature, the reaction solution was purified by C-18 reverse phase chromatography to obtain the title compound 14 (20 mg, yield 57.1%).

MS m/z (ESI): 383.3 [M+H] ⁺ .

¹ H NMR(400MHz,DMSO-d6):δ11.94(s,1H),8.17(s,1H),7.98-7.92(m,1H),7.38(s,1H),6.93(d,J=7.6 Hz,1H),6.28(s,1H),5.00(d,J=6.9Hz,1H),3.87(s,2H),3.58(h,J=6.7Hz,1H),3.09-2.99(m,1H ),2.5-2.40(m,1H),2.22(s,3H),2.00(t,J＝7.8Hz,1H),1.90(ddd,J＝13.3,8.0,5.0Hz,1H),1.74(s, 2H), 1.65 (s, 1H), 1.03 (dd, J=6.6, 2.1 Hz, 6H).

Example 15

(1R,3S)-3-(3-((5-(cyanomethyl)-4-methylpyridin-2-yl)amino)-1H-pyrazol-5-yl)isopropylcarbamate cyclopentyl ester

Synthesis Reference Example 14, compound 15 can be synthesized by changing the relevant raw materials.

MS m/z (ESI): 383.3 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ): δ11.82(s, 1H), 9.09(s, 1H), 7.99(s, 1H), 7.16(s, 1H), 6.94(d, J＝7.8Hz ,1H),6.05(s,1H),5.00(d,J=7.3Hz,1H),3.87(s,2H),3.58(h,J=6.6Hz,1H),3.11-2.97(m,1H) ,2.45(dd,J=14.2,7.4Hz,1H),2.25(s,3H),2.01(q,J=8.5,6.9Hz,1H),1.96-1.83(m,1H),1.73(q,J = 7.3, 6.2Hz, 2H), 1.59 (d, J = 14.3Hz, 1H), 1.04 (d, J = 6.6Hz, 6H).

Example 16

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-(5-{[5-(formamidomethyl)pyrazin-2-yl]amino}-2H-pyrazole-3- base) cyclopentyl ester

Compound 16 was prepared according to the synthesis steps of Example 11 by replacing the corresponding raw materials.

MS m/z (ESI): 388.4 [M+H] ⁺ .

¹ H NMR(400MHz,DMSO):δ11.93(s,1H),9.44(s,1H),8.51(s,1H),8.02(d,J＝1.4Hz,1H),7.43(s,1H) ,6.93(s,2H),6.15(s,1H),5.00(s,1H),3.63-3.54(m,1H),3.46(s,1H),3.10-3.01(m,1H),2.06-1.83 (m, 3H), 1.81-1.55 (m, 4H), 1.04 (d, J=6.4Hz, 6H).

Example 17

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-(5-{[6-(cyanomethyl)pyrazin-2-yl]amino}-2H-pyrazol-3-yl ) cyclopentyl ester

first step

2-(6-Bromopyrazin-2-yl)-2-cyanoacetic acid-2-methylpropan-2-yl ester 17b

First, 2,6-dibromopyrazine (1 g, 4.2 mmol) was dissolved in N, N-dimethylformamide (40 ml), followed by the addition of tert-butyl 2-cyanoacetate (1.2 g, 8.4 mmol), Cesium carbonate (3.4g, 10.5mmol) was reacted at room temperature for 4 hours. The crude product is obtained after extraction, drying and concentration, which can be directly used in the next reaction.

MS m/z (ESI): 298.2 [M+H] ⁺ .

second step

(6-Bromopyrazin-2-yl)acetonitrile 17c

Firstly, the resulting crude product 2-(6-bromopyrazin-2-yl)-2-cyanoacetic acid-2-methylpropan-2-yl ester was dissolved in dichloromethane (20ml), then an excess of tris Fluoroacetic acid (4ml), react at room temperature for 1 hour. The crude product was obtained after extraction, drying and concentration, and (6-bromopyrazin-2-yl)acetonitrile 17c (250 mg, 30%) was separated by normal phase column chromatography.

MS m/z (ESI): 198.2 [M+H] ⁺ .

third step

(Propan-2-ylamino)methane acid-(1R,3S)-3-(5-{[6-(cyanomethyl)pyrazin-2-yl]amino}-1-(2-methylpropane -2-yl)pyrazol-3-yl)cyclopentyl ester 17e

Under nitrogen atmosphere, (6-bromopyrazin-2-yl) acetonitrile (50mg, 0.25mmol), (1R,3S)-3-(5-amino-1-(tert-butyl)-1H-pyridine Azol-3-yl) cyclopentyl isopropyl carbamate (77mg, 0.25mmol, prepared by the method disclosed in the patent application "WO 2020/157652 A2"), XantPhos (8.7mg 0.015mmol), Pd2dba3 ( 11.4mg, 0.0125mmol), cesium carbonate (81.2mg, 0.25mmol) were dissolved in dioxane (2mL), and microwaved at 120°C for 1 hour. The reaction solution was cooled to room temperature, filtered, and the filtrate was collected, concentrated under reduced pressure and separated from the normal phase to obtain the title compound 17e (38 mg, yield 36%).

MS m/z (ESI): 426.4 [M+H] ⁺ .

the fourth step

(Propan-2-ylamino)methane acid-(1R,3S)-3-(5-{[6-(cyanomethyl)pyrazin-2-yl]amino}-2H-pyrazol-3-yl ) cyclopentyl ester 17

(Propan-2-ylamino)methane acid-(1R,3S)-3-(5-{[6-(cyanomethyl)pyrazin-2-yl]amino}-1-(2-methylpropane -2-yl)pyrazol-3-yl)cyclopentyl ester (38mg, 0.09mmol) was dissolved in 3mL formic acid and reacted at 100°C for 1 hour. After cooling to room temperature, the reaction solution was purified by C-18 reverse phase chromatography to obtain the title compound 636 (9 mg, yield 28%).

MS m/z (ESI): 370.2 [M+H] ⁺ .

Example 18

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-(5-{[6-(formamidomethyl)pyrazin-2-yl]amino}-2H-pyrazole-3- base) cyclopentyl ester

Compound 18 was prepared according to the synthesis steps of Example 11 by replacing the corresponding raw materials.

¹ H NMR(400MHz,DMSO):δ11.93(s,1H),9.45(s,1H),8.51(s,1H),8.02(d,J＝1.5Hz,1H),7.43(s,1H) ,6.93(s,1H),6.88(d,J=8.3Hz,1H),6.15(s,1H),5.00(d,J=8.4Hz,1H),3.46(s,2H),3.41-3.36( m, 1H), 3.13-3.01 (m, 1H), 2.09-1.84 (m, 3H), 1.82-1.54 (m, 3H), 1.01 (d, J=6.6Hz, 6H).

Example 19

(Propan-2-ylamino)methane acid-(1R,3S)-3-(5-{[6-(cyanomethyl)-1,2-diazacyclohexyl-3-yl]amino} -2H-pyrazol-3-yl)cyclopentyl ester

Compound 19 was prepared according to the synthetic steps of Example 11 by replacing the corresponding raw materials.

MS m/z (ESI): 370.2 [M+H] ⁺ .

Example 20

(But-2-ylamino)methane-(1R,3S)-3-(5-{[5-(formamidomethyl)pyrazin-2-yl]amino}-2H-pyrazole-3- base) cyclopentyl ester

Compound 20 was prepared according to the synthesis steps of Example 11 by replacing the corresponding raw materials.

MS m/z (ESI): 402.2 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO): δ11.94(s,1H),9.54(s,1H),8.45(s,1H),7.83(s,1H),7.54(s,1H),7.00(s, 1H), 6.94(d, J=7.9Hz, 1H), 6.18(s, 1H), 5.01(s, 1H), 3.63-3.53(m, 1H), 3.48(s, 2H), 3.09-3.00(m ,1H),2.49-2.41(m,1H),2.07-1.96(m,3H),1.94-1.85(m,1H),1.78-1.57(m,3H),1.06-1.04(m,3H),1.03 -1.02(m,3H).

Example 21

(Propan-2-ylamino)methane acid-(1R,3S)-3-[5-({4-[Dimethyl(oxyethylene)-5-methylphosphoryl]phenyl}amino)-2H- Pyrazol-3-yl]cyclopentyl ester

Compound 21 was prepared according to the synthetic steps of Example 11 by replacing the corresponding raw materials.

MS m/z (ESI): 405.2 [M+H] ⁺ .

Example 22

(1R,3S)-3-(3-((6-cyanopyridazin-3-yl)amino)-1H-pyrazol-5-yl)cyclopentyl((S)-sec-butyl)aminomethyl Ester

first step

(1R,3S)-3-(1-(tert-butyl)-5-((6-cyanopyridazin-3-yl)amino)-1H-pyrazol-3-yl)cyclopentyl ((S )-sec-butyl)carbamate

(1R,3S)-3-(5-Amino-1-(tert-butyl)-1H-pyrazol-3-yl)cyclopentyl((S)-sec-butyl)carbamate (84mg, 0.26mmol) was dissolved in N,N-dimethylformamide (10mL), sodium hydride (13mg, 0.31mmol) was added at 0°C, stirred at 0°C for 0.5 hours, then 6-chloropyridazine-3-formazine was added Nitrile 22a (36mg, 0.26mmol), after the reaction was completed, the reaction solution was washed with 1mL of water, extracted with 20mL of ethyl acetate, washed with 20mL of saturated aqueous sodium chloride solution, the organic phase was collected, 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 chromatography to obtain the title compound 22c (40 mg, yield 40.9%).

MS m/z (ESI): 426.2 [M+H] ⁺ .

second step

(1R,3S)-3-(3-((6-cyanopyridazin-3-yl)amino)-1H-pyrazol-5-yl)cyclopentyl((S)-sec-butyl)aminomethyl Ester

Compound 22 was prepared according to the synthesis steps of Example 11 by replacing the corresponding raw materials.

MS m/z (ESI): 370.2 [M+H] ⁺ .

Example 23

(1R,3S)-3-(3-((2-fluoro-4-(N-(deuteromethyl)sulfonyl)phenyl)amino)-1H-pyrazol-5-yl)cyclopentyliso Propyl carbamate

first step

4-Bromo-3-fluoro-N-(deuteromethyl)benzenesulfonamide 23b

Deuteromethylamine hydrochloride (154mg, 2.2mmol) was dissolved in 15mL of dichloromethane, triethylamine (557mg, 5.52mmol), 4-dimethylaminopyridine (45mg, 0.37mmol) were added to the system, 0 Add 4-bromo-3-fluorobenzenesulfonyl chloride (500mg, 1.84mmol) at ℃ and stir overnight at room temperature. After the reaction, the reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel chromatography to obtain the title compound 23b (145 mg, yield 29.2%).

MS m/z (ESI): 270.9 [M+H] ⁺ .

second step

(1R,3S)-3-(1-(tert-butyl)-5-((2-fluoro-4-(N-(deuteromethyl)sulfonyl)phenyl)amino)-1H-pyrazole- 3-yl) cyclopentyl isopropyl carbamate 23c

Compound 23c was prepared according to the synthetic procedure of Example 11 by replacing the corresponding raw materials. (56 mg, yield 75.9%).

MS m/z (ESI): 499.2 [M+H] ⁺ .

third step

(1R,3S)-3-(3-((2-fluoro-4-(N-(deuteromethyl)sulfonyl)phenyl)amino)-1H-pyrazol-5-yl)cyclopentyliso Propyl carbamate 23

Compound 23 was prepared according to the synthesis steps of Example 11 by replacing the corresponding raw materials. (19 mg, yield 38.2%).

MS m/z (ESI): 443.2 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d6): δ11.99(s, 1H), 8.68(d, J=1.6Hz, 1H), 8.23-8.19(m, 1H), 7.47-7.43(m, 2H), 7.18(s,1H),6.96(d,J=7.2Hz,1H),5.84(s,1H),5.01(d,J=4.0Hz,1H),3.61-3.56(m,1H),3.09-3.05 (m, 1H), 2.06-1.60 (m, 6H), 1.03 (d, J=6.4Hz, 6H).

Example 24

(1R,3S)-3-(3-((4-(N-(Deuteromethyl)sulfonyl)phenyl)amino)-1H-pyrazol-5-yl)cyclopentylisopropylaminomethyl Ester

Compound 24 (19 mg, yield 38.2%) was obtained by replacing the corresponding raw materials according to the synthetic procedure of Example 23.

MS m/z (ESI): 425.2 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ): δ11.91(s, 1H), 8.93(s, 1H), 7.55(d, J=8.8Hz, 2H), 7.41(d, J=8.0Hz, 2H ),7.02(s,1H),6.94(d,J=7.2Hz,1H),5.71(s,1H),5.00(d,J=2.8Hz,1H),3.61-3.56(m,1H),3.08 -3.04 (m, 1H), 2.06-1.61 (m, 6H), 1.03 (d, J=6.4Hz, 6H).

Example 25

(1R,3S)-3-(3-((6-(N-(methyl-d3)aminosulfonyl)pyridin-3-yl)amino)-1H-pyrazol-5-yl)cyclopentyliso Propyl carbamate

first step

5-Bromo-N-(methyl-d3)pyridine-2-sulfonamide 25b

At room temperature, deuterated methylamine hydrochloride (210mg, 3mmol) and DIEA (650mg, 5mmol) were dissolved in anhydrous dichloromethane (10mL), and after clarification, 5-bromopyridine-2-sulfonyl chloride ( 250mg, 1mmol), continue to stir, TLC monitors until the end of the reaction, add water (10mL) to wash once, and saturated brine (10mL) to wash once, collect the organic phase and dry it over anhydrous sodium sulfate, spin to a small volume and directly normalize Column chromatography gave the title compound 25b (200 mg, yield 82%).

MS m/z (ESI): 253.9 [M+H] ⁺ .

second step

(1R,3S)-3-(1-(tert-butyl)-5-((6-(N-(methyl-d3)aminosulfonyl)pyridin-3-yl)amino)-1H-pyrazole- 3-yl) cyclopentyl isopropyl carbamate 25c

Under nitrogen atmosphere, 5-bromo-N-(methyl-d3)pyridine-2-sulfonamide (25.3 mg, 0.10 mmol), (1R,3S)-3-(5-amino-1-(tert- Butyl)-1H-pyrazol-3-yl)cyclopentylisopropylcarbamate (30.8mg, 0.10mmol, prepared by the method disclosed in the patent application "WO 2020/157652 A2"), Pd2( dba)3 (9.1mg, 0.01mmol), Xantphos (11.4mg, 0.02mmol), CS2CO3 (96.2mg, 0.30mmol) were dissolved in 1,4-dioxane (2mL), and reacted under microwave conditions at 80°C 2 Hour. The reaction solution was cooled to room temperature, filtered, and the filtrate was collected, concentrated under reduced pressure and separated from the normal phase to obtain the title compound 25c (16 mg, yield 32%).

MS m/z (ESI): 482.3 [M+H] ⁺ .

third step

(1R,3S)-3-(3-((6-(N-(methyl-d3)aminosulfonyl)pyridin-3-yl)amino)-1H-pyrazol-5-yl)cyclopentyliso Propyl carbamate 25

25c (16mg, 0.03mmol) was dissolved in 2mL formic acid and reacted at 80°C for 12 hours. After cooling to room temperature, the reaction solution was purified by C-18 reverse phase chromatography to obtain the title compound 25 (2.5 mg, yield 19.6%).

MS m/z (ESI): 426.2 [M+H] ⁺ .

Example 26

(1R,3S)-3-(3-((6-(N-Methylaminosulfonyl)pyridin-3-yl)amino)-1H-pyrazol-5-yl)cyclopentyl ((S)- sec-butyl) carbamate

The title compound 26 was prepared according to the synthesis procedure of Example 25 by replacing the corresponding raw materials.

MS m/z (ESI): 437.2 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ): δ12.01(s, 1H), 9.23(s, 1H), 8.58(d, J=2.6Hz, 1H), 8.45(s, 1H), 7.99(d ,J=8.6Hz,1H),7.74(d,J=8.7Hz,1H),7.33(q,J=5.0Hz,1H),6.89(d,J=8.3Hz,1H),5.73(s,1H ),5.00(s,1H),3.12-3.03(m,1H),2.47(d,J=4.8Hz,3H),2.01(dp,J=12.9,7.0,6.3Hz,2H),1.91(ddd, J=12.9,8.4,4.3Hz,1H),1.78-1.70(m,2H),1.61(t,J=14.2Hz,1H),1.47(d,J=7.3Hz,1H),1.41-1.30(m , 1H), 1.02 (d, J=6.6Hz, 3H), 0.83 (dt, J=23.5, 6.9Hz, 3H).

Example 27

(1R,3S)-3-(3-((5-(N-methylaminosulfonyl)pyridin-2-yl)amino)-1H-pyrazol-5-yl)cyclopentylisopropylaminomethyl Ester

The title compound 27 was prepared according to the synthesis procedure of Example 25 by replacing the corresponding raw materials.

MS m/z (ESI): 423.2 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ): δ12.03(s, 1H), 9.84(s, 1H), 8.44(d, J=2.5Hz, 1H), 7.82(dd, J=9.0, 2.6Hz ,1H),7.34(s,1H),7.26(d,J=5.5Hz,1H),6.94(d,J=7.7Hz,1H),6.18(s,1H),5.01(s,1H),3.63 -3.53(m,1H),3.07(t,J=8.9Hz,1H),2.41(d,J=4.8Hz,3H),2.00(p,J=6.9,6.5Hz,1H),1.74(d, J=9.0Hz, 1H), 1.62(s, 1H), 1.46(t, J=7.1Hz, 1H), 1.25(m, 4H), 1.04(dd, J=6.6, 2.2Hz, 3H).

Example 28

(1R,3S)-3-(3-((6-cyanopyridin-3-yl)amino)-1H-pyrazol-5-yl)cyclopentylisopropylcarbamate

Compound 28 was prepared according to the synthetic steps of Example 11 by replacing the corresponding raw materials.

MS m/z (ESI): 355.2 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ): δ12.08(s, 1H), 9.43(s, 1H), 8.58(d, J=2.6Hz, 1H), 7.96(dd, J=8.8, 2.7Hz ,1H),7.77(d,J=8.7Hz,1H),6.94(d,J=7.8Hz,1H),5.75(s,1H),5.00(dd,J=6.6,3.2Hz,1H),3.58 (dt,J=9.8,5.0Hz,1H),3.12-3.01(m,1H),2.09-1.97(m,1H),2.00-1.84(m,1H),1.74(q,J=7.6,4.7Hz , 2H), 1.61 (s, 1H), 1.24 (d, J=3.7Hz, 1H), 1.04 (dd, J=6.6, 2.3Hz, 6H).

Example 29

(1R,3S)-3-(3-((5-cyanopyridin-3-yl)amino)-1H-pyrazol-5-yl)cyclopentylisopropylcarbamate

Compound 29 was prepared according to the synthetic steps of Example 11 by replacing the corresponding raw materials.

MS m/z(ESI): 355.2[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ ): δ11.97(s, 1H), 9.15(s, 1H), 8.70(d, J=2.5Hz, 1H), 8.34-8.27(m, 2H), 6.94 (d,J=7.7Hz,1H),5.70(d,J=1.9Hz,1H),5.00(s,1H),3.62-3.53(m,1H),3.12-3.02(m,1H),2.09- 1.84(m,1H),1.73(s,1H),1.61(s,1H),1.46(s,1H),1.24(s,2H),1.04(d,J=6.5Hz,6H).

Example 30

(1R,3S)-3-(3-((4-(methylsulfonyl)phenyl)amino)-1H-pyrazol-5-yl)cyclopentylisopropylcarbamate

Compound 30 was prepared according to the synthesis steps of Example 11 by replacing the corresponding raw materials.

MS m/z (ESI): 407.2 [M+H] ⁺ .

¹ H NMR(400MHz,DMSO-d ₆ ):δ11.95(s,1H),9.06(s,1H),7.67(d,J=8.8Hz,2H),7.50-7.43(m,2H),6.94 (d,J=6.8Hz,1H),5.72(s,1H),5.01(s,1H),3.57(s,1H),3.08(s,3H),2.03(dt,J=11.0,7.5Hz, 1H), 1.99-1.84(m, 1H), 1.73(d, J=8.5Hz, 2H), 1.61(s, 1H), 1.24(s, 1H), 1.04(dd, J=6.7, 2.2Hz, 6H ).

Example 31

(1R,3S)-3-(3-((5-((methylsulfonyl)methyl)pyridin-2-yl)amino)-1H-pyrazol-5-yl)cyclopentylisopropylaminomethyl Ester

first step

2-Chloro-5-((methylthio)methyl)pyridine 31b

Dissolve 2-chloro-5-(chloromethyl)pyridine (1000mg, 6.2mmol) in 95% ethanol (10mL) at room temperature, add sodium methylthiolate (700mg, 10mmol) slowly after clarification, and continue stirring, After TLC monitoring to the end of the reaction, water (10 mL) was added to wash once, saturated brine (10 mL) was washed once, the organic phase was collected and dried over anhydrous sodium sulfate, the solvent was spin-off under reduced pressure, and compound 31b was obtained after column chromatography ( 800 mg, yield 74%).

MS m/z (ESI): 174.0 [M+H] ⁺ .

second step

2-Chloro-5-((methylsulfonyl)methyl)pyridine 31c

Dissolve 2-chloro-5-((methylthio)methyl)pyridine (800mg, 4.5mmol) in dichloromethane (10mL) at room temperature, add m-CPBA (1100mg, 6.75mmol) slowly after clarification , continue to stir, TLC monitors until the end of the reaction, filter the precipitate, add water (10mL) to wash once, and saturated brine (10mL) to wash once, collect the organic layer, dry over anhydrous sodium sulfate, and spin off the solvent under reduced pressure , Compound 31c (800 mg, yield 85%) was obtained after column chromatography.

MS m/z (ESI): 206.0 [M+H] ⁺ .

third step

(1R,3S)-3-(1-(tert-butyl)-5-((5-((methylsulfonyl)methyl)pyridin-2-yl)amino)-1H-pyrazol-3-yl) Cyclopentyl isopropyl carbamate 31d

Under nitrogen atmosphere, 2-chloro-5-((methylsulfonyl)methyl)pyridine (20.6mg, 0.10mmol), (1R,3S)-3-(5-amino-1-(tert-butyl )-1H-pyrazol-3-yl)cyclopentylisopropylcarbamate (30.8mg, 0.10mmol, prepared by the method disclosed in the patent application "WO 2020/157652 A2"), Pd ₂ (dba ) ₃ (9.1mg, 0.01mmol), Xantphos (11.4mg, 0.02mmol), CS ₂ CO ₃ (96.2mg, 0.30mmol) were dissolved in 1,4-dioxane (2mL), and microwaved at 80°C React for 2 hours. The reaction solution was cooled to room temperature, filtered, and the filtrate was collected, concentrated under reduced pressure and separated from the normal phase to obtain the title compound 31d (5 mg, yield 10%).

MS m/z (ESI): 478.3 [M+H] ⁺ .

the fourth step

(1R,3S)-3-(3-((5-((methylsulfonyl)methyl)pyridin-2-yl)amino)-1H-pyrazol-5-yl)cyclopentylisopropylaminomethyl Ester 31

(1R,3S)-3-(1-(tert-butyl)-5-((5-((methylsulfonyl)methyl)pyridin-2-yl)amino)-1H-pyrazol-3-yl ) cyclopentyl isopropyl carbamate (5 mg, 0.01 mmol) was dissolved in 1 mL of formic acid, and reacted at 80° C. for 12 hours. After cooling to room temperature, the reaction solution was purified by C-18 reverse phase chromatography to obtain the title compound 31 (1.2 mg, yield 28.3%).

MS m/z (ESI): 422.2 [M+H] ⁺ .

Example 32

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-{5-[(4-formamidophenyl)amino]-1H-pyrazol-3-yl}cyclopentyl ester

Compound 32 was prepared according to the synthesis steps of Example 8 by replacing the corresponding raw materials.

MS m/z (ESI): 372.2 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ): δ11.81(s, 1H), 8.66(s, 1H), 7.71(d, J=8.8Hz, 2H), 7.61(s, 1H), 7.30(d ,J=8.3Hz,2H),6.94(d,J=6.9Hz,2H),5.67(s,1H),5.08–4.95(m,1H),3.76-3.51(m,1H),3.16-2.95( m,1H),2.47-2.40(m,1H),2.09-1.97(m,1H),1.93-1.85(m,1H),1.80-1.64(m,2H),1.64-1.58(m,1H), 1.08-1.00(m,6H).

Example 33

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-[5-({2-fluoro-4-[(methylamino)dioxylidene-6-sulfanyl]phenyl}ammonia Base)-1H-pyrazol-3-yl]cyclopentyl ester

Compound 33 was prepared according to the synthetic steps of Example 8 by replacing the corresponding raw materials.

MS m/z (ESI): 440.2 [M+H] ⁺ .

¹ H NMR(400MHz,DMSO-d ₆ ):δ11.99(s,1H),8.69(s,1H),8.21(t,J=8.7Hz,1H),7.54-7.41(m,2H),7.26 –7.15(m,1H),6.94(d,J＝7.8Hz,1H),5.84(s,1H),5.03-4.98(m,1H),3.60-3.55(m,1H),3.14-3.00(m ,1H),2.49-2.43(m,1H),2.39(d,J＝5.1Hz,3H),2.10-1.84(m,2H),1.76-1.65(m,2H),1.64-1.54(m,1H ), 1.04 (d, J=6.8, 2.1 Hz, 6H).

Example 34

(Propan-2-ylamino)methane-(1R,3S)-3-{5-[(4-{[(2-hydroxyethyl)amino]dioxylidene-6-thio}phenyl) Amino]-1H-pyrazol-3-yl}cyclopentyl ester

Compound 34 was prepared according to the synthesis steps of Example 8 by replacing the corresponding raw materials.

MS m/z (ESI): 452.2 [M+H] ⁺ .

Example 35

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-{5-[(5-cyanopyrimidin-2-yl)amino]-1H-pyrazol-3-yl}cyclopentyl ester

Compound 35 was prepared according to the synthesis steps of Example 8 by replacing the corresponding raw materials.

MS m/z (ESI): 356.2 [M+H] ⁺ .

¹ H NMR(400MHz,DMSO-d ₆ ):δ12.21(s,1H),9.74(s,1H),8.94(s,2H),6.94(s,1H),5.79(s,1H),5.00 (s,1H),3.62-3.55(m,1H),3.13-3.04(m,1H),2.48-2.38(m,1H),2.09-1.94(m,1H),1.97-1.86(m,1H) , 1.77-1.69 (m, 2H), 1.64-1.59 (m, 1H), 1.04 (d, J=6.5Hz, 6H).

Example 36

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-{5-[(5-formamidopyrazin-2-yl)amino]-2H-pyrazol-3-yl}ring Amyl esters

Compound 36 was prepared according to the synthesis steps of Example 11 by replacing the corresponding raw materials and reagents.

MS m/z (ESI): 374.2 [M+H] ⁺ .

¹ H NMR(400MHz,DMSO-d ₆ ):δ12.12(s,1H),10.14(s,1H),8.67(s,1H),8.47(s,1H),7.77(s,1H),7.40 (s,1H),6.95(d,J＝7.7Hz,1H),6.29(s,1H),5.04-4.99(m,1H),3.65-3.52(m,1H),3.13-3.04(m,1H ),2.48–2.42(m,1H),2.09-1.99(m,1H),1.98-1.83(m,1H),1.78-1.71(m,2H),1.65-1.60(m,1H),1.04(d , J=6.6, 2.4Hz, 6H).

Example 37

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-(5-{[5-(formamidomethyl)pyridin-2-yl]amino}-2H-pyrazole-3- base) cyclopentyl ester

Compound 37 was prepared according to the synthetic steps of Example 11 by replacing the corresponding raw materials and reagents.

MS m/z (ESI): 387.3 [M+H] ⁺ .

Example 38

(Propan-2-ylamino)methane acid-(1R,3S)-3-[5-({4-[Dioxyylidene(propionylamino)-6-thio]phenyl}amino)-2H- Pyrazol-3-yl]cyclopentyl ester

Compound 38 was prepared according to the synthetic steps of Example 11 by replacing corresponding raw materials and reagents.

MS m/z (ESI): 464.2 [M+H] ⁺ .

Example 39

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-{5-[(4-cyanopyrimidin-2-yl)amino]-2H-pyrazol-3-yl}cyclopentyl ester

Compound 39 was prepared according to the synthesis steps of Example 11 by replacing the corresponding raw materials and reagents.

MS m/z (ESI): 356.2 [M+H] ⁺ .

Example 40

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-(5-{[1-(methyldioxyylidene-6-thio)hexahydropyridin-4-yl]ammonia Base}-2H-pyrazol-3-yl)cyclopentyl ester

first step

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-(5-{[1-(methyldioxyylidene-6-thio)hexahydropyridin-4-yl]ammonia Base}-1-(2-methylprop-2-yl)pyrazol-3-yl)cyclopentyl ester 40b

Compound 1-N-methylsulfonyl-4-piperidone (178 mg, 1.0 mmol), 1a (310 mg, 1.0 mmol) was dissolved in dichloromethane (10 mL), added to dry 4A molecular sieves and stirred at room temperature for 1 hour. Add glacial acetic acid (1 drop), sodium cyanoborohydride (124 mg, 2.0 mmol) and react overnight at room temperature. Water (30 mL) was added to the reaction liquid, and extracted with dichloromethane (20 mL×2). The organic phases were combined and dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated in vacuo to obtain the crude compound. The crude product was purified by C18 to obtain compound 40b (234 mg, 0.5 mmol, yield: 50%). MS(ESI)m/z 470.3[M+H]

second step

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-(5-{[1-(methyldioxyylidene-6-thio)hexahydropyridin-4-yl]ammonia Base}-2H-pyrazol-3-yl)cyclopentyl ester 40

Compound 40 was prepared according to the synthesis steps of Example 8 by replacing the corresponding raw materials.

MS m/z (ESI): 414.3 [M+H] ⁺ .

Example 41

(Propan-2-ylamino)methane-(1R,3S)-3-[5-(1,2-diazacyclohexyl-3-ylamino)-2H-pyrazol-3-yl]ring Amyl esters

Compound 41 was prepared according to the synthetic steps of Example 11 by replacing the corresponding raw materials.

MS m/z (ESI): 331.2 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ): δ11.94(s, 1H), 9.51(s, 1H), 8.59(d, 1H), 7.54(d, J=9.1Hz, 1H), 7.44-7.35 (m,1H),6.95(d,J＝7.7Hz,1H),6.22(s,1H),5.05-4.98(m,1H),3.65-3.53(m,1H),3.11-3.02(m,1H ),2.49-2.43(m,1H),2.07-1.97(m,1H),1.98-1.83(m,1H),1.80-1.67(m,2H),1.65-1.60(m,1H),1.04(d , J=6.7, 2.0Hz, 6H).

Example 42

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-{5-[(5-cyano-4-methylpyridin-2-yl)amino]-2H-pyrazol-3-yl } Cyclopentyl ester

Compound 42 was prepared according to the synthesis steps of Example 11 by replacing the corresponding raw materials.

MS m/z (ESI): 369.2 [M+H] ⁺ .

biological evaluation

The following further describes and explains the present disclosure in combination with test examples, but these examples are not meant to limit the scope of the present disclosure.

Test Example 1. Test of the Inhibitory Activity of the Compounds of the Disclosure on Ovarian Cancer Cells (OVCAR3)

Table 1. Experimental materials and instruments

1. Experimental steps

Ovarian cancer cells OVCAR3 were cultured in RPMI 1640 with 10% FBS in a 37%, 5% _CO2 incubator. On the first day, cells were plated in 96-well plates at a concentration of 2500 cells/well and incubated overnight in an incubator. Compound treatment was performed on the second day, the highest concentration of compound treatment was 10 uM, 3-fold dilution, 9 concentrations, and the final concentration of DMSO was 0.1%. After the cells continued to be cultured in the incubator for 7 days, the cell viability was tested using the Celltiter Glo assay kit (Promega), and the test method was consistent with the operation method provided by the kit. Data were processed and _IC50 calculated using GraphPad Prism 8.

Calculation formula Y=Bottom+(Top-Bottom)/(1+10^((LogIC50-X)*HillSlope)).

X: log value of compound concentration; Y: % inhibition.

The IC ₅₀ (nM) of the disclosed compound on OVCAR3, A≤400nM, 400nM<B≤600nM, 600nM<C≤1000nM.

Table 2. IC ₅₀ (nM) for OVCAR3 of Compounds of the Disclosure

Example number IC ₅₀ (nM) the Example 1 A 248.5 Example 2 C 741 Example 4 A 102 Example 5 A 36.5 Example 6 A 84.6 Example 7 C 593 Example 8 B 446 Example 9 A 330 Example 10 A 175 Example 11 B 495 Example 12 A 86 Example 17 A 32 Example 23 A 259 Example 24 A 140 Example 26 A 69 Example 29 B 431 Example 33 A 253 Example 34 A 290 Example 39 B 562 EX32 / 2964 EX402 / 9249 EX563 / 1359

Note: Compounds EX32, EX402 and EX563 were synthesized respectively according to Example 32, Example 402 and Example 563 of WO2022174031A.

Test Example 2. Detection of the activity of the disclosed compounds on cyclin-dependent kinases (CDK1, 2, 9).

2.1 Experimental materials and instruments (see Table 3)

Table 3. Experimental materials and instruments

2.2 Experimental steps

Compound dilutions were transferred to each well of the assay plate using an Echo 550 (784075, Greiner). Seal the assay plate and centrifuge the assay plate at 1000g for 1 min; prepare 2x enzyme in 1x kinase buffer ((40mM Tris-HCl, pH 7.4, 20mM MgCl2, 0.1mg/ml BSA, 50uM DTT) by dissolving 2.5 μl of 2x enzyme Add to 384-well assay plate, centrifuge the plate at 1000g for 30s, and let it stand at room temperature for 10 minutes. Prepare 2x substrate and ATP mix in 1x kinase buffer, add 2.5μl 2x substrate and ATP mix to start the reaction. Centrifuge the plate at 1000g Centrifuge for 30 seconds, seal the assay plate, and react at room temperature for 1 hour. Add 4 μl ADP-Glo reagent, incubate at room temperature for 40 minutes, then add 8 μl kinase detection reagent, and incubate at room temperature for 40 minutes.

The luminescent signal of each well was read on an Envision 2104 plate reader.

Percent inhibition was calculated as follows: Percent inhibition = 100 - (cmpd signal - Ave_PC signal) / (Ave_VC signal - Ave_PC signal) x 100. cmpd signal: compound signal; Ave_PC signal: positive control signal average; Ave_VC signal: vector group signal average.

_IC50s were calculated using GraphPad 8.0 by fitting the percent inhibition values and the logarithm of the compound concentration to a non-linear regression (dose response - variable slope). Y=Bottom+(Top-Bottom)/(1+10^(( _LogIC50 -X)*Slope)), where X: logarithm of inhibitor concentration; Y: % inhibition.

Table 4. IC ₅₀ values of compounds of the present disclosure to CDK1/B, CDK2/Cyclin E, CDK9/Cyclin T1

Note: Compounds EX32, EX402 and EX563 were synthesized according to Example 32, Example 402 and Example 563 of WO2022174031A, respectively.

Test example 3. the pharmacokinetic test of embodiment 8 of the present invention and embodiment 9

Taking Balb/c nude mouse as the test animal, the LC/MS/MS method was used to determine the drug concentration in the blood plasma at different times after the oral administration of Example 8 and Example 9. Study the pharmacokinetic behavior of the compound of the present invention in Balb/c nude mouse body, and evaluate its pharmacokinetic characteristics.

3.1 Experimental animals

Two healthy female Balb/c nude mice were purchased from Jizui Yaokang Company.

3.2 Drug preparation

Weigh an appropriate amount of sample, and use 5% DMSO/40% PEG400/55% saline as a solvent to prepare a 7.5 mg/mL solution.

3.3 Administration

Two healthy female Balb/c nude mice were intragastrically administered after overnight fasting, the dose was 75mg/kg, and the administration volume was 10mL/kg.

Blood was collected before administration and at 0.25, 0.5, 1, 2, 4, 8, and 24 hours after administration, placed in a heparinized anticoagulant test tube, centrifuged to separate plasma, and stored at low temperature. The content of the test compound in plasma was determined by LC/MS/MS method. Plasma samples were pretreated to precipitate proteins for analysis.

3.4 Results of pharmacokinetic parameters

Table 5. The pharmacokinetic parameters of the disclosed compounds are as follows:

Claims (23)

A compound represented by formula I or a pharmaceutically acceptable salt thereof,

Wherein, said ^R1 and ^R2 are each independently selected from hydrogen, alkyl, haloalkyl, hydroxyalkyl, cycloalkyl or heterocycloalkyl, said alkyl, haloalkyl, hydroxyalkyl, ring Alkyl or heterocycloalkyl is optionally substituted by one or more substituents independently selected from R ^1A ; The R ^1A is selected from halogen, hydroxyl, alkyl, alkoxy, haloalkyl, haloalkoxy, heterocycloalkyl or NR'(R"), and the alkyl, alkoxy, haloalkyl, Haloalkoxy or heterocycloalkyl is optionally substituted by one or more substituents independently selected from R ⁵ ; Or said R ¹ and R ² form a 3 to 7-membered heterocycloalkyl group with the nitrogen atom they are jointly connected to, and said heterocycloalkyl group is optionally substituted by one or more substituents independently selected from R ^1B ; The R ^1B is selected from halogen, cyano, hydroxyl, amino, alkyl or alkoxy; The R ³ is selected from H, COR' or

The ring A is selected from 5 to 6 membered aryl, 5 to 6 membered heteroaryl, cycloalkyl or heterocycloalkyl; The ^R4 are each independently selected from hydrogen, halogen, cyano, nitro, hydroxyl, alkenyl, alkynyl, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, Heterocycloalkyl, cycloalkyloxy, heterocycloalkyloxy, aryl, heteroaryl, SR', SOR', SO ₂ R', -NHSO ₂ R', SO ₂ NR'(R") , NR'(R"), COR', -NHCOR', COOR', CONR'(R"), -(P=O)R'(R"),

Or oxo, the alkenyl, alkynyl, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocycloalkyl, cycloalkyloxy, heterocycloalkane Oxy, aryl or heteroaryl is optionally substituted by one or more substituents independently selected from R ^4A ; Alternatively, two adjacent R ⁴ together form ring A optionally substituted by one or more independently selected from R ^4A

Described R ^4A is selected from halogen, cyano, nitro, hydroxyl, alkenyl, alkynyl, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocycloalkyl, Cycloalkyloxy, heterocycloalkyloxy, aryl, heteroaryl, SR', SOR', SO ₂ R', -NHSO ₂ R', SO ₂ NR'(R"), NR'(R ”), COR', -NHCOR', COOR', CONR'(R"), -(P=O)R'(R"), or

The alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocycloalkyl, cycloalkyloxy, heterocycloalkyloxy, aryl or heteroaryl are any is optionally substituted by one or more substituents independently selected from R ⁸ ; Said X is independently selected from O or S; The R ⁵ is selected from halogen, hydroxyl, alkyl, cyano, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl or NR'(R"); The R ^and R ^are each independently selected from hydrogen, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl or heterocycloalkyl; The ^R is selected from halogen, hydroxyl, cyano, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl or NR'(R"); The R' and R" are each independently selected from hydrogen or alkyl; Said n is selected from 1, 2, 3, 4 or 5. The compound shown in formula I according to claim 1 or a pharmaceutically acceptable salt thereof, which is a compound shown in formula II or a pharmaceutically acceptable salt thereof,

Wherein, said R ¹ , R ² , R ³ , R ⁴ and n are as defined in claim 1 respectively. The compound shown in formula I according to claim 1 or a pharmaceutically acceptable salt thereof, which is a compound shown in formula III or IV or a pharmaceutically acceptable salt thereof,

Wherein, said R ¹ , R ² , R ⁴ and n are as defined in claim 1 respectively. The compound represented by formula I or a pharmaceutically acceptable salt thereof according to any one of claims 1-3, wherein, Ring A is selected from phenyl, five-membered nitrogen-containing heteroaryl or six-membered nitrogen-containing heteroaryl ; preferably pyridyl, pyrimidinyl, piperidinyl, imidazolyl, pyrrolyl or pyrazolyl; most preferably pyridyl. The compound represented by formula I or a pharmaceutically acceptable salt thereof according to any one of claims 1-3, wherein Ring A is I-1,

The G ¹ , G ² , G ³ , G ⁴ and G ⁵ are each independently selected from CR ⁴ or N, wherein at least one of G ¹ , G ² , G ³ , G ⁴ and G ⁵ is CR ⁴ ; or Ring A is I-2,

The G ¹ , G ² , G ³ and G ⁴ are each independently selected from CR ⁴ or N or NR ⁴ , and the R ⁴ is as defined in claim 1. The compound represented by formula I or a pharmaceutically acceptable salt thereof according to claim 5, wherein ring A is selected from I-3 to 1-14,

Wherein, G ¹ , G ² , G ³ , G ⁴ and G ⁵ are each independently selected from CR ⁴ , and said R ⁴ is as defined in claim 1. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein said ^R is selected from hydrogen, alkyl, haloalkyl, hydroxyalkyl, cycloalkyl or heterocycloalkyl ; Preferably hydrogen, C _1-6 alkyl, C _1-6 haloalkyl, C _1-6 hydroxyalkyl, 3-7 membered cycloalkyl or 3-7 membered heterocycloalkyl, more preferably hydrogen, C _1-3 alkyl, C _1-3 haloalkyl; most preferably hydrogen. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, wherein said ^R is selected from alkyl, haloalkyl, hydroxyalkyl, cycloalkyl or heterocycloalkyl, and The alkyl, haloalkyl, hydroxyalkyl, cycloalkyl or heterocycloalkyl are optionally substituted by one or more substituents independently selected from R ^1A , and the R ^1A is selected from haloalkyl, alkane Oxygen, haloalkyl, haloalkoxy, heterocycloalkyl or NR'(R"), said alkyl, alkoxy, haloalkyl, haloalkoxy, heterocycloalkyl are optionally replaced by one or more R ⁵ replaces; Preferably, said R ² is selected from C _1-6 alkyl, C _1-6 haloalkyl, C _1-6 hydroxyalkyl, 3 to 7-membered cycloalkyl or 3 to 7-membered heterocycloalkyl , the C _1-6 alkyl, C _1-6 haloalkyl, C _1-6 hydroxyalkyl, 3 to 7 membered cycloalkyl or 3 to 7 membered heterocycloalkyl are optionally replaced by one or more Substituents independently selected from R ^1A are substituted, and said R ^1A is selected from halogen or C _1-6 alkyl; More preferably, said ^R is selected from methyl, ethyl, isopropyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, said cyclopropyl, cyclobutyl, cyclopentyl The group is substituted by one or more R ^1A , and the R ^1A is selected from methyl, ethyl, isopropyl; Particularly preferably, said R ² is selected from isopropyl or methylcyclopropyl. According to the compound represented by formula I or its pharmaceutically acceptable salt according to any one of claims 1 to 2, 4 to 8, wherein, said ^R is selected from

Wherein X is O, the ^R6 and ^R7 are each independently selected from hydrogen, _C1-6 alkyl, preferably hydrogen, methyl, ethyl. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, wherein each ^R is independently selected from hydrogen, cyano, halogen, hydroxyl, alkyl, alkoxy, haloalkyl, Haloalkoxy, Hydroxyalkyl, Cycloalkyl, Heterocycloalkyl, Cycloalkyloxy, Heterocycloalkyloxy, Heteroaryl, SO ₂ R', -NHSO ₂ R', COR', -NHCOR ', COOR', CONR'(R"), or

The alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocycloalkyl, heteroaryl are optionally substituted by one or more R ^4A ; Described R ^4A is selected from halogen, cyano, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocycloalkyl, cycloalkyloxy, heterocycloalkyloxy group, SO ₂ R', -NHSO ₂ R', COR', -NHCOR', COOR', CONR'(R"), or

Said R' and R" are as defined in claim 1 respectively. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, wherein each of said R ⁴ is independently selected from hydrogen, cyano or halogen; Alternatively, each of said R ⁴ is independently selected from hydrogen, COR', -NHSO ₂ R', SO ₂ R',

Alternatively, each R ⁴ is independently selected from hydrogen, alkyl, hydroxyalkyl, the alkyl is optionally substituted by one or more R ^4A , and the R ^4A is selected from halogen, cyano, alkoxy, SO ₂ R',

Alternatively, each R ^is independently selected from hydrogen, alkoxy, heterocycloalkyloxy, preferably C _1-6 alkoxy or 3 to 7 membered heterocycloalkyloxy; Said R' and R" are as defined in claim 1 respectively. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 11, wherein said R ¹ is hydrogen, and said R ² is selected from isopropyl or methylcyclopropyl. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 12, said R' and R" are each independently selected from hydrogen or C _1-6 alkyl, preferably hydrogen or methyl, ethyl . According to any one of claims 3 to 8, 10 to 13, the compound shown in formula IV or a pharmaceutically acceptable salt thereof, the ring A is phenyl; The R ⁴ are each independently selected from hydrogen, halogen, -NHSO ₂ R' or SO ₂ NR'(R"); The R ¹ and R ² are each independently selected from hydrogen, alkyl, haloalkyl; The R' and R" are selected from hydrogen or C _1-6 alkyl; The n is selected from 1, 2, 3, 4 or 5; Preferably, said R ⁴ are each independently selected from hydrogen, halogen or SO ₂ NR'(R"); The R ¹ and R ² are each independently selected from hydrogen or C _1-6 alkyl; The R' and R" are selected from hydrogen or C _1-3 alkyl; Said n is selected from 1, 2, 3, 4 or 5. According to the compound shown in formula IV or its pharmaceutically acceptable salt according to any one of claims 3 to 14, said

selected from

preferred

A compound or a pharmaceutically acceptable salt thereof as shown below, which is selected from:

An isotope substitution of the compound according to any one of claims 1 to 16; preferably, the isotope substitution is deuterium atom substitution. A pharmaceutical composition comprising at least one therapeutically effective amount of the compound or pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 16, the isotope substitution described in claim 17 and a pharmaceutically acceptable excipient. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 16, the isotope substitution described in claim 17 or the pharmaceutical composition described in claim 18 is used in the preparation for prevention and/or treatment of Use in medicine for protein-dependent kinase-related diseases, the protein-dependent kinase is preferably CDK2, and the protein-dependent kinase-related diseases are preferably cell proliferation diseases, cancer or immune diseases. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 16, the isotope substitution described in claim 17 or the pharmaceutical composition described in claim 18 is used in the preparation for prevention and/or treatment of The purposes in the medicine of cell cyclin related disease, described cell cycle protein is preferably cell cycle protein E, more preferably cell cycle protein E1, cell cycle protein E2, described cell cycle protein related disease is preferably cell proliferative disease, cancer or immune disease. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 16, the isotope substitution described in claim 17 or the pharmaceutical composition described in claim 18 is used in the preparation for preventing and/or treating cancer The purposes of the medicine, described cancer is selected from breast cancer, ovarian cancer, prostate cancer, melanoma, brain tumor, esophageal cancer, gastric cancer, liver cancer, pancreatic cancer, colorectal cancer, lung cancer, kidney cancer, skin cancer, adult Glioblastoma, neuroblastoma, or sarcoma. A method for preparing the compound shown in formula III according to any one of claims 3 to 9, 11 to 16 or a pharmaceutically acceptable salt thereof or the isotope substitution described in claim 17, which comprises formula V The step of removing the amino protecting group PG of the compound under acidic conditions,

Among them, R ¹ , R ² , R ⁴ , and n are respectively as defined in claim 1, and PG is an amino protecting group, preferably tert-butyl, acetyl, trifluoroacetyl, trityl, benzyl, methyl Acyl, most preferably tert-butyl. A compound shown in formula V,

Among them, R ¹ , R ² , R ⁴ , and n are respectively as defined in claim 1, and PG is an amino protecting group, preferably tert-butyl, acetyl, trifluoroacetyl, trityl, benzyl, methyl Acyl, most preferably tert-butyl.