TW202325280A - An aminopyrazole derivative, preparation method and use thereof - Google Patents

Abstract

The disclosure provides an aminopyrazole derivative, preparation method and use thereof. In particular, the present disclosure provides a compound shown in formula I or a pharmaceutically available salt thereof. The compounds shown in formula I can be used as cyclin dependent kinase inhibitors to prevent and/or treat diseases related to protein dependent kinase or cyclin. The groups in general formula I are defined in the specification.

Description

A kind of aminopyrazole derivative and its preparation method and application

The disclosure relates to an aminopyrazole derivative and its preparation method and use, which belong to the field of medicine.

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 important role in division and proliferation. Cyclin-dependent kinase catalytic units are activated by regulatory subunits called cyclins. At least 16 mammalian cell cycle proteins have been identified (Annu. Rev. Pharmacol. Toxicol. (1999) 39:295-312). Cyclin B/CDK1, cyclin A/CDK2, cyclin E/CDK2, cyclin D/CDK4, cyclin D/CDK6 and 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 CDK4/6 monotherapy or combined with endocrine therapy in hormone receptor positive advanced breast cancer, such as palbociclib, ribociclib (ribociclib) and abemaciclib have been approved in combination with aromatase inhibitors 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 palbociclib and abemaciclib have been approved in combination with fulvestrant for the treatment of hormone receptor (HR)-positive, postmenopausal women after disease progression on endocrine therapy. Human epidermal growth factor receptor 2 (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 separately identified and discovered a series of inhibitors that selectively inhibit CDK 2 for the treatment of diseases such as cancer, such as Selixili (Seliciclib), Dinaciclib, etc., but in order to achieve better cancer treatment effects and better meet market demand, it is still necessary to develop a new generation of selective CDK2 inhibitors with high efficiency and low toxicity.

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

Wherein, the R ^and R are ^each independently selected from hydrogen, alkyl, haloalkyl, hydroxyalkyl, cycloalkyl or heterocycloalkyl, and the alkyl, haloalkyl, hydroxyalkyl, cycloalkyl Or heterocycloalkyl is optionally substituted by one or more substituents independently selected from R ^1A ;

The R ^1A is selected from halogen, hydroxy, alkyl, alkoxy, haloalkyl, haloalkoxy, heterocycloalkyl or NR'(R"), the alkyl, alkoxy, haloalkyl, halo Alkoxy 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 each 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;

或側氧基，該烯基、炔 基、烷基、烷氧基、鹵烷基、鹵烷氧基、羥烷基、環烷基、雜環烷基、環烷基氧基、雜環烷基氧基、芳基、雜芳基視需要被一個或多個各自獨立地選自R^4A的取代基取代； The R ^{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 pendant oxy, 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 ;

或

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

or

;

，該烷基、烷氧基、鹵烷基、鹵烷氧 基、羥烷基、環烷基、雜環烷基、環烷基氧基、雜環烷基氧基、芳基、雜芳基視需要被一個或者多個R⁸取代； The R ^4A is selected from halogen, cyano, nitro, hydroxy, 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 Replaced by one or more R ⁸ as required;

The X is independently selected from O or S;

The R ^is selected from halogen, hydroxy, 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 an alkyl group;

The 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.

，該G¹、G²、G³、G⁴和G⁵各自 獨立地選自CR⁴或N，其中，G¹、G²、G³、G⁴和G⁵中至少有一個為CR⁴，該R⁴如式I所示化合物中定義。 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 represented by formula I.

，該G¹、G²、G³和G⁴各自獨立 地選自CR⁴或N或NR⁴，該R⁴如式I所示化合物中定義。 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.

，該G²、G³、G⁴和G⁵各自獨立地選 自CR⁴，該R⁴如式I所示化合物中定義。 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.

，該G¹、G³、G⁴和G⁵各自獨立地選 自CR⁴，該R⁴如式I所示化合物中定義。 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 shown in formula I.

，該G¹、G²、G⁴和G⁵各自獨立地選 自CR⁴，該R⁴如式I所示化合物中定義。 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.

，該G³、G⁴和G⁵各自獨立地選自 CR⁴，該R⁴如式I所示化合物中定義。 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.

，該G²、G⁴和G⁵各自獨立地選自CR⁴， 該R⁴如式I所示化合物中定義。 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.

，該G²、G³和G⁴各自獨立地選自CR⁴， 該R⁴如式I所示化合物中定義。 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.

，該G¹、G³和G⁴各自獨立地選自 CR⁴，該R⁴如式I所示化合物中定義。 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.

，該G²、G³和G⁵各自獨立地選自 CR⁴，該R⁴如式I所示化合物中定義。 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.

，該G¹、G³和G⁵各自獨立地選自 CR⁴，該R⁴如式I所示化合物中定義。 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.

，該G³和G4各自獨立地選自CR⁴， 該R⁴如式I所示化合物中定義。 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 G4 are each independently selected from CR ⁴ , and the R ⁴ is as defined in the compound shown in formula I.

，該G⁴為CR⁴，該R⁴如式I所示化 合物中定義。 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.

，該G³為CR⁴，該R⁴如式I所示化 合物中定義。 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 .

，G¹、G²、G³、G⁴和G⁵選自CR⁴，該

選自

、

或

。 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

,

or

.

為

。 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

.

，G²、G³、G⁴和G⁵各自獨立地選自CR⁴。 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 .

，診

選自

、

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

, diagnosis

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

.

，G¹、G³、G⁴和G⁵各自獨立地選自CR⁴。 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 .

或

。 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

or

.

，G²、G⁴和G⁵各自獨立地選自CR⁴，該R⁴如式I所示化合物中定義。 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 shown in formula I.

，G³、G⁴和G⁵各自獨立地選自CR⁴。 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

.

，G²、G⁴和G⁵各自獨立地選自CR⁴，該R⁴如式I所示化合物中定義。 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 shown in formula I.

，G²、G³、G⁴各自獨立地選自CR⁴，該R⁴如式I所示化合物中定義。 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.

，G¹、G³、G⁴各自獨立地選自CR⁴。 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 .

或

。 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

or

.

，G²、G³、G⁵各自獨立地選自CR⁴，該R⁴如式I所示化合物中定義。 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.

，G¹、G³和G⁵各自獨立地選自CR⁴，該R⁴如式I所示化合物中定義。 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 shown in formula I.

，G³、G⁴各自獨立地選自CR⁴，該R⁴如式I所示化合物中定義。 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 the R ⁴ is as defined in the compound shown in formula I.

，G⁴選自CR⁴，該R⁴如式I所示化合物中定義。 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 the R ⁴ is as defined in the compound shown in formula I.

，G³選自CR⁴，該R⁴如式I所示化合物中定義。 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 the R ⁴ is as defined in the compound shown in formula I.

，其中環A的定義如本文任一方案所述。 In an alternative embodiment, two adjacent R ⁴ together with ring A ^form a

, wherein ring A is as defined 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 the ^R is selected from hydrogen, alkyl, haloalkyl, hydroxyalkyl , 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 the 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, the 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 R ¹ is hydrogen.

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

In an optional embodiment, the present disclosure provides a compound represented by formula I, II, III, IV or a pharmaceutically acceptable salt thereof, wherein, the R ² is selected from C _1-6 alkyl, C _1-6 halogen Alkyl, 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 substituted by one or more R ^1A ; the 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, the R ² is selected from methyl, ethyl, isopropyl, third Butyl, cyclopropyl, cyclobutyl, cyclopentyl, the cyclopropyl, cyclobutyl, cyclopentyl are substituted by one or more R ^1A , the R ^1A is selected from methyl, ethyl, isopropyl .

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

，其中X為O，該R⁶和R⁷各自獨立地選自氫、C_1-6烷基。 In an optional embodiment, the present disclosure provides a compound represented by formula I, II or a pharmaceutically acceptable salt thereof, wherein, the R ³ 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為O，該R⁶和R⁷各自獨立地選自氫、甲基、乙基。 In an optional embodiment, the present disclosure provides a compound represented by formula I, II or a pharmaceutically acceptable salt thereof, wherein, the R ³ is selected from

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

、

、

或

。 In an optional embodiment, the present disclosure provides compounds represented by formulas I and II or pharmaceutically acceptable salts thereof, wherein the ^R3 is preferably

,

,

or

.

，該烯基、炔基、烷基、烷氧基、鹵烷基、鹵烷氧基、羥烷基、環烷基、雜環烷基、環烷基氧基、雜環烷基氧基、芳基、雜芳基視需要被一個或多個R^4A取代，其中，R^4A如式I所示的化合物中定義。 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 base, 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, Aryl and heteroaryl are optionally substituted by one or more R ^4A , wherein R ^4A is as defined in the compound shown in formula I.

或側氧基，該烯基、炔基、烷基、烷氧基、鹵烷基、鹵烷氧基、羥烷基、環烷基、雜環烷基、環烷基氧基、雜環烷基氧基、芳基、雜芳基視需要被一個或多個R^4A取代，其中，R^4A如式I所示的化合物中定義。 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 base, SR', SOR', SO ₂ R', -NHSO ₂ R', SO ₂ NR'(R"), NR'(R"), COR', -NHCOR', COOR', CONR'(R" ), -(P=O)R'(R"),

Or pendant oxy, 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 a compound represented by formulas I, II, III, IV or a pharmaceutically acceptable salt thereof, wherein R ⁴ is a pendant oxy group.

，該烷基、烷氧基、鹵烷基、鹵烷氧基、羥烷基、環烷基、雜環烷基、雜芳基視需要被一個或多個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 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 ;

。 The 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 a compound represented by formula I, II, III, IV or a pharmaceutically acceptable salt thereof, wherein each R ⁴ is independently selected from hydrogen, cyano or halogen.

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, COR', -NHSO ₂ R ', SO ₂ R'

。 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 R ⁴ is independently selected from hydrogen, alkyl, hydroxyalkyl, The alkyl is optionally substituted by one or more R ^4A 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 R ⁴ is independently selected from hydrogen, alkoxy, heterocycloalkane Baseoxy, 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 R ¹ is hydrogen.

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

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

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 formulas I, II, III, IV or its pharmaceutically acceptable salt or its isotope substitution and at least one pharmaceutically acceptable carrier body, diluent or excipient. In some embodiments, the unit dose of the pharmaceutical composition is 0.001 mg-1000 mg.

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

The present disclosure also provides a method for preventing and/or treating a patient suffering from a cell cycle protein-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-associated disease or cyclin-associated 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 Carcinoma, lung cancer, kidney cancer, skin cancer, glioblastoma, neuroblastoma, sarcoma.

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

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 formulas I, II, III, IV described in the present disclosure or its druggable The 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 Carcinoma, skin cancer, glioblastoma, neuroblastoma, sarcoma.

The present disclosure provides a therapeutically effective amount of the compounds represented by the formulas I, II, III, and IV described in the present disclosure or their pharmaceutically acceptable salts or their isotope substitutions or the aforementioned pharmaceutical compositions for the prevention 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 represented by the formula I, II, III, IV or its pharmaceutically acceptable salt or its isotope substitution or the aforementioned pharmaceutical composition in preparation for use in the present disclosure. 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 the compound represented by formula I, II, III, IV of the present disclosure or its pharmaceutically acceptable salt or its isotope substitution or the aforementioned pharmaceutical composition in the preparation of 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, adult Glioma, neuroblastoma, sarcoma.

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

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 amine 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 tertiary butyl, acetyl, trifluoroacetyl, triphenyl Methyl, benzyl, 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 tertiary butyl, acetyl, trifluoroacetyl, triphenyl Methyl, benzyl, formyl.

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

The disclosed compound has a good inhibitory effect on cyclin-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 achieve <10 nM inhibitory activity against CDK2/Cyclin E. Other embodiments provide compounds that achieve <5 nM inhibitory activity against CDK2/Cyclin E. Other embodiments provide compounds that achieve <1 nM inhibitory activity against CDK2/Cyclin E.

On the other hand, the disclosed compounds also have good inhibitory effect on CDK2/Cyclin A. Some embodiments provide compounds that achieve <500 nM inhibitory activity against CDK2/Cyclin A. another The compounds provided by some embodiments achieve <200nM inhibitory activity on 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 achieve <50 nM inhibitory activity against CDK2/Cyclin A. 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 disclosed herein 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, compounds of the present disclosure 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 the present 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-lactimine, pyrazolyl isomerisation.

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 amine group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or alkali, and then by conventional methods known in the art Methods The diastereoisomers were resolved and the pure enantiomers were recovered. Furthermore, the separation of enantiomers and diastereomers is usually accomplished by the use of chromatography using a chiral stationary phase, optionally combined with chemical derivatization methods (e.g. amines to amines formate).

The disclosure also includes 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 at least 1000 times greater than its natural abundance (which is 0.015%) (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.

”表示未指定構型，即如果化學結構中存在手性異構體，鍵“

”可以為“

”或“

”，或者同時 包含“

”和“

”兩種構型。雖然為簡便起見將全部上述結構式畫成某些異構體形式，但是本發明可以包括所有的異構體，如互變異構體、旋轉異構體、幾何異構體、非對映異構體、外消旋體和對映異構體。 In the chemical structure of the compound described in the present invention, the bond "

"indicates that no configuration is specified, 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 structural formulas are drawn as certain isomer forms for simplicity, the present invention may include all isomers, such as tautomers, rotamers, geometric isomers isomers, diastereomers, racemates and enantiomers.

”並未指定構型，即可以為Z構型或E構型，或者同時包含兩種構型。 In the chemical structures of the compounds described in this disclosure, the bond "

" does not specify the configuration, it can be the Z configuration or the 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 living body, facilitate the absorption of the active ingredient and thus 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 this 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 allow 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, second-butyl, n-pentyl, 1,1-dimethylpropane base, 1,2-dimethylpropyl, 2,2-dimethylpropyl and its 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, hydroxyl, pendant oxygen , 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 by halogen, hydroxy, nitro, cyano or amine.

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 may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from alkoxy, halo, haloalkyl, haloalkoxy, hydroxy , hydroxyalkyl, 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 , hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl.

The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, and the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 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, hydroxyl, pendant Oxygen, cyano, amino, C _1-6 alkyl, C _1-6 alkoxy, 3 to 6 membered cycloalkyl or 3 to 6 membered heterocycloalkyl, the alkyl, alkoxy, cycloalkane The radical or heterocycloalkyl group is optionally substituted with halogen, hydroxy, nitro, cyano or amine.

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 contain 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably contain 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:

和

等。

and

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, hydroxyl, pendant oxygen, 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 depending on Requires substitution by halogen, hydroxy, nitro, cyano or amine.

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, hydroxyl, pendant oxygen, cyano, amine, C _1-6 alkyl, C _1-6 alkoxy, 3 to 7 membered cycloalkyl or 3 to 7 membered heterocycloalkyl, the alkyl, alkoxy, cycloalkyl or heterocycloalkyl optionally Substituted by halogen, hydroxy, nitro, cyano or amine.

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 can 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 ring Alkyl, 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-membered heterocycloalkylthio, the alkoxy, cycloalkyl, heterocycloalkyl, cycloalkoxy, heterocyclyloxy, alkylthio, cycloalkylthio, heterocycloalkylthio are optionally Halogen, hydroxyl, 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 containing 1 to 6 carbon atoms, non-limiting examples include methylene (-CH ₂ -), ethylidene (such as -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, "(extend) alkoxy", "(extend) alkenylene", "(extend) alkenyloxy", "(extend) cycloalkyl", "(extend) heterocycloalkane The definition of "group" is like "(alkylene)".

The term "aryl" refers to a 6 to 14 membered all carbon monocyclic or fused polycyclic (i.e. rings sharing adjacent pairs of carbon atoms) group having a conjugated pi electron system, preferably 6 to 12 members, e.g. phenyl 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, hydroxyl, pendant oxygen, nitro, cyano, 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, C _{3 -8} cycloalkenyloxy, 5 to 6 membered aryl or heteroaryl, the 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. Heteroaryl is preferably 6 to 12 members, more preferably 5 or 6 members. For example. Non-limiting examples thereof include: imidazolyl, furyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, isoxazolyl, pyrrolyl, tetrazolyl, pyridyl, pyrimidinyl , Thiadiazole, pyrazinyl, triazolyl, indazolyl, benzimidazolyl,

,

,

wait.

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

Heteroaryl 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, hydroxyl, cyano, amino, C _{1- 6} alkyl or C _1-6 alkoxy.

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 "side oxygen" refers to a =O substituent.

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

"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.

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 examples of the present disclosure that do not indicate specific conditions are usually 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 carried out with a Bruker AVANCE-400 nuclear magnetic instrument, and the determination solvent is deuterated dimethyl sulfide (DMSO-d ₆ ), deuterated chloroform (CDCl ₃ ), deuterated methanol (CD ₃ OD), and the internal standard is four Methylsilane (TMS).

MS was measured with 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 1200 LC high pressure liquid chromatography (Ultimate XB-C18 3.0*150mm chromatography column or Xtimate C182.1*30mm chromatography column).

Chiralpak IC-3 100×4.6mm ID, 3um, Chiralpak AD-3 150×4.6mm ID, 3um, Chiralpak AD-3 50×4.6mm ID, 3um, Chiralpak AS-3 150×4.6mm were used for chiral HPLC analysis and determination ID, 3um, Chiralpak AS-3 100×4.6mm ID, 3 μm , ChiralCel OD-3 150×4.6mm ID, 3um, Chiralcel OD-3 100×4.6mm ID, 3 μm , ChiralCel OJ-H 150× 4.6mmI.D., 5um, Chiralcel OJ-3 150×4.6mmI.D., 3um chromatographic 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 preparation column used 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 used 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 Chemicals and other 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 a Parr 3916EKX hydrogenator and Qinglan QL-500 hydrogen generator or HC2-SS hydrogenator.

The hydrogenation reaction is usually evacuated 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 temperature of the reaction 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 the reaction, the eluting agent system and the developing agent system of the column chromatography that the purified compound adopts and the thin-layer chromatography, the solvent The volume ratio is adjusted according to the polarity of the compound, and 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-(Third-butyl)-5-((4-cyanopyridin-2-yl)amino)-1H-pyrazol-3-yl)cyclopentyliso Propyl carbamate 1b

(1R,3S)-3-(5-Amino-1-(tertiary-butyl)-1H-pyrazol-3-yl)cyclopentyl isopropyl carbamate (40mg, 0.13mmol , prepared by the method disclosed in the patent application "WO 2020/157652 A2"), 2-chloroisonicotinonitrile (20mg, 0.14mmol), palladium acetate (5.8mg, 0.03mmol), 1,1'-binaphthyl - 2.2'-Diphenylphosphine (32.3mg, 0.05mmol), cesium carbonate (84.5mg, 0.26mmol) and 1,4-dioxane (1.5mL) 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-pyrazole-3- Formic acid (1.5 mL) was added to cyclopentyl 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): δ 11.99(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 ester

first step

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

Under nitrogen atmosphere, 2-(6-chloropyridin-3-yl)acetonitrile (20mg, 0.13mmol), (1R,3S)-3-(5-amino-1-(tertiary-butyl )-1H-pyrazol-3-yl)cyclopentyl isopropyl carbamate (40.4 mg, 0.13 mmol, prepared by the method disclosed in the patent application "WO 2020/157652 A2"), palladium acetate ( 5.9 mg, 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 Ester 2

(1R,3S)-3-(1-(tertiary-butyl)-5-((5-(cyanomethyl)pyridin-2-yl)amino)-1H-pyrazol-3-yl ) cyclopentyl isopropyl carbamate (40 mg, 0.09 mmol) was dissolved in 6 mL 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): δ 11.84(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) Cyclopentyl 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, and 2,4-dichloropyridine (300 mg, 2.02 mmol) was added to the reaction solution in batches , stirred at 100°C for 2 hours, cooled to room temperature, added 50ml of water to the reaction solution, extracted with ethyl acetate, combined the organic phases, washed the organic phases, dried, concentrated under reduced pressure, and the residue was reverse-phased with C-18 Purification by chromatography afforded the title compound 3b (120 mg, 32.8% yield).

MS m/z (ESI): 181.2 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO): δ 9.58(s, 1H), 8.58(d, J =5.5Hz, 1H), 8.38(s, 1H), 8.11(d, J = 2.0Hz, 1H), 7.95( dd, J =5.6,1.9Hz,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)cyclopentyl isopropyl carbamate 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-(Tertiary-butyl)-1H-pyrazol-3-yl)cyclopentylisopropylcarbamate (25.6mg, 0.08mmol, using the method disclosed in patent application "WO 2020/157652 A2" prepared), palladium acetate (3.7mg, 0.02mmol), 1,1'-binaphthyl-2,2'-bisdiphenylphosphine (10.3mg, 0.02mmol) dissolved in 1,4-dioxane (2mL )middle. Add cesium carbonate (67.7mg, 0.21mmol) and react at 80°C for 2 hours. The reaction solution was cooled to room temperature, filtered, the filtrate was collected, and concentrated under reduced pressure to obtain the title compound 3c (30mg, 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) Cyclopentyl isopropyl carbamate 3

(1R,3S)-3-(5-((4-(1H-1,2,4-triazol-1-yl)pyridin-2-yl)amino)-1-(third-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): δ 12.21-11.66 (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)methanoic acid-(1R,3S)-3-[5-({4-[(methylamino)dioxylidene-6-thio]phenyl}amino) -2H-pyrazol-3-yl]cyclopentyl ester

Compound 4 was prepared according to the synthetic 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-pyrazole-3 -yl)cyclopentyl ester

first step

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

First, 2-bromo-5-chloropyrazine 5a (500mg, 2.6mmol) was dissolved in N,N-dimethylformamide (20ml), followed by the addition of tert-butyl 2-cyanoacetate (733mg, 5.2 mmol), 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 (150mg, 29%) was obtained by normal phase column chromatography.

MS m/z (ESI): 198.2 [M+H] ⁺ .

third step

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-(5-{[5-(cyanomethyl)pyrazin-2-yl]amino}-1-(2-methyl propyl-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 -pyrazol-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), and cesium carbonate (81.2mg, 0.25mmol) were dissolved in dioxane (2mL), and reacted under microwave conditions 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 in 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-pyrazole-3 -yl) cyclopentyl ester 5

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-(5-{[5-(cyanomethyl)pyrazin-2-yl]amino}-1-(2- Methylpropan-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): δ 12.00(s, 1H), 9.70(s, 1H), 8.55(s, 1H), 8.11(d, J =1.4Hz, 1H), 6.94(d, J =7.7 Hz,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(d J =6.7Hz, 6H).

Example 6

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

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

MS m/z (ESI): 356.4 [M+H] ⁺ .

¹ H NMR (400MHz,DMSO- d ₆ ): δ 12.21(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)methanoic acid-(1R,3S)-3-{5-[(6-cyano-1,2-diazacyclohexyl-3-yl)amino]-2H -pyrazol-3-yl}cyclopentyl ester

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

MS m/z (ESI): 356.4 [M+H] ⁺ .

Example 8

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

first step

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

Under nitrogen atmosphere, 4-bromobenzenesulfonamide (23.5mg, 0.10mmol), (1R,3S)-3-(5-amino-1-(tertiary-butyl)-1H-pyrazole- 3-yl) cyclopentyl isopropyl carbamate (30.8 mg, 0.10 mmol, prepared by the method disclosed in the patent application "WO 2020/157652 A2"), XantPhos Pd G3 (9.5 mg, 0.01 mmol) 1. 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 in normal phase to obtain the title compound 8b (40mg, yield 85%).

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

third step

(1R,3S)-3-(3-((4-sulfonylphenyl)amino)-1H-pyrazol-5-yl) cyclopentylisopropylcarbamate8

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 ): δ 11.87(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)methanoic acid-(1R,3S)-3-[5-({4-[(amino)dioxylidene-6-thio]-2fluorophenyl}amino )-2H-pyrazol-3-yl]cyclopentyl ester

first step

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-(5-{[4-(aminodioxylidene-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)methanoic acid-(1R,3S)-3-[5-({4-[(amino)dioxylidene-6-thio]-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 ): δ 11.96(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}methanoic acid-(1R,3S)-3-(5-{[4-(aminodioxylidene-6-thio)-2-fluoro Phenyl]amino}-2H-pyrazol-3-yl)cyclopentyl ester

first step

{[(2S)-Butan-2-yl]amino}methanoic acid-(1R,3S)-3-(5-{[4-(aminodioxylidene-6-thio)-2-fluoro Phenyl]amino}-1-(2-methylpropan-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}methanoic acid-(1R,3S)-3-(5-{[4-(aminodioxylidene-6-thio)-2-fluoro Phenyl]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 ): δ 11.95(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 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 direct normal phase column chromatography, 11b was obtained (180 mg, 72% yield).

second step

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

Under nitrogen atmosphere, 5-bromo-N-methylpyridine-2-sulfonamide (25.1mg, 0.10mmol), (1R,3S)-3-(5-amino-1-(tertiary-butyl base)-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), microwave at 80°C The reaction was carried out for 2 hours. The reaction solution was cooled to room temperature, filtered, and the filtrate was collected, concentrated under reduced pressure and separated in 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-Methylaminosulfonyl)pyridin-3-yl)amino)-1H-pyrazol-5-yl)cyclopentylisopropyl Base Urethane 11

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

MS m/z (ESI): 423.2 [M+H] ⁺ .

Example 12

{[(2S)-Butan-2-yl]amino}methanoic acid-(1R,3S)-3-[5-({6-[(Ethylamino)dioxylidene-6-thio] Pyridin-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): δ 12.00(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-pyrazole-5 -yl)cyclopentyl (S)-second 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). React overnight at 105°C, cool down to room temperature, concentrate the reaction solution under reduced pressure, add 30 mL of water, extract with ethyl acetate, combine the organic phases, wash the organic phase with saturated brine, dry, and concentrate under reduced pressure to obtain the residue 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, washed with saturated brine, dried, and concentrated under reduced pressure. Separation and purification by silica gel column chromatography gave the title compound 13c (514.7 mg, yield 70%).

MS m/z (ESI): 201.1 [M+H] ⁺ .

third step

2-(5-((1-(tertiary butyl)-3-((1S,3R)-3-((((S)-second butyl)aminoformyl)oxy)cyclopentyl Base)-1H-pyrazol-5-yl)amino)pyrazin-2-yl)ethyl acetate 13d

2-(5-Chloropyrazin-2-yl)ethyl acetate (500mg, 2.5mmol), (1R,3S)-3-(5-amino-1-(tert-butyl)-1H-pyridine Azol-3-yl) cyclopentyl ((S)-second butyl) carbamate (805mg, 2.5mmol, using 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), nitrogen Reflux under protection. After the reaction was completed, the reaction solution was lowered to room temperature, 20ml of water was added, extracted with ethyl acetate, the organic phase was combined, and the organic phase was washed with saturated brine, dried, and concentrated under reduced pressure. Separation and purification by column chromatography gave the title compound 13d (850.5 mg, yield 70%).

MS m/z (ESI): 487.5 [M+H] ⁺ .

the fourth step

2-(5-((1-(tertiary butyl)-3-((1S,3R)-3-((((S)-second butyl)aminoformyl)oxy)cyclopentyl Base)-1H-pyrazol-5-yl)amino)pyrazin-2-yl)acetic acid 13e

The 2-(5-((1-(tertiary butyl)-3-((1S,3R)-3-((((S)-second butyl)aminoformyl)oxy) ring Amyl)-1H-pyrazol-5-yl)amino)pyrazin-2-yl)ethyl acetate 13d (500mg, 1.03mmol) was dissolved in tetrahydrofuran (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-(tertiary butyl)-5-((5-(2-(deuteromethylamino)-2-oxoethyl)pyrazin-2-yl)amine Base)-1H-pyrazol-3-yl)cyclopentyl ((S)-second 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, and the organic phases were combined. The organic phase was washed with 1M hydrochloric acid, 1M aqueous sodium hydroxide solution and saturated saline, dried, concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography. The title compound 13f was obtained (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-pyrazole-5 -yl)cyclopentyl-(S)-second butyl carbamate 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- Isoxazole 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, drop to At 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-pyrazole-3 -yl) cyclopentyl isopropylcarbamate 14d

14c (35.8mg, 0.21mmol), (1R,3S)-3-(5-amino-1-(tertiary-butyl)-1H-pyrazol-3-yl)cyclopentylisopropyl Carbamate (60mg, 0.19mmol) was 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): δ 11.94(s,1H),8.17(s,1H),7.98-7.92(m,1H),7.38(s,1H),6.93(d, J =7.6Hz ,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 =6.6,2.1Hz,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 ): δ 11.82(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-yl)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): δ 11.93(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-pyrazole-3 -yl)cyclopentyl ester

first step

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

First, 2,6-dibromopyrazine (1g, 4.2mmol) was dissolved in N,N-dimethylformamide (40ml), then tert-butyl 2-cyanoacetate (1.2g, 8.4mmol) was added ), cesium carbonate (3.4g, 10.5mmol), 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): 298.2 [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) was reacted at room temperature for 1 hour. After extraction, drying, and concentration, the crude product was obtained, which was separated by normal phase column chromatography to obtain (6-bromopyrazin-2-yl)acetonitrile 17c (250mg, 30%).

MS m/z (ESI): 198.2 [M+H] ⁺ .

third step

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-(5-{[6-(cyanomethyl)pyrazin-2-yl]amino}-1-(2-methyl Propyl-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 -pyrazol-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), and cesium carbonate (81.2mg, 0.25mmol) were dissolved in dioxane (2mL), and reacted under microwave conditions 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 in 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)methanoic acid-(1R,3S)-3-(5-{[6-(cyanomethyl)pyrazin-2-yl]amino}-2H-pyrazole-3 -yl)cyclopentyl ester 17

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-(5-{[6-(cyanomethyl)pyrazin-2-yl]amino}-1-(2-methyl Propan-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-yl)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): δ 11.93(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)methanoic acid-(1R,3S)-3-(5-{[6-(cyanomethyl)-1,2-diazepan-3-yl]amine Base}-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)methanoic acid-(1R,3S)-3-(5-{[5-(formamidomethyl)pyrazin-2-yl]amino}-2H-pyrazole -3-yl)cyclopentyl ester

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

MS m/z (ESI): 402.2 [M+H] ⁺ .

¹ H NMR (400MHz,DMSO): δ 11.94(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)methanoic 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)-second butyl) Urethane

first step

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

(1R,3S)-3-(5-Amino-1-(tertiary butyl)-1H-pyrazol-3-yl)cyclopentyl ((S)-second butyl)carbamic acid Dissolve the ester (84mg, 0.26mmol) in N,N-dimethylformamide (10mL), add sodium hydride (13mg, 0.31mmol) at 0°C, stir at 0°C for 0.5 hours, then add 6-chloropyridium Oxyzine-3-carbonitrile 22a (36 mg, 0.26 mmol), after the reaction, the reaction solution was washed with 1 mL of water, extracted with 20 mL of ethyl acetate, washed with 20 mL of saturated aqueous sodium chloride solution, and the organic phase was collected and dried over anhydrous sodium sulfate. Filter and collect the filtrate. 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)-second butyl) Urethane

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)cyclopentyl Isopropyl carbamate

first step

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

Deuteromethylamine hydrochloride (154mg, 2.2mmol) was dissolved in 15mL of dichloromethane, and triethylamine (557mg, 5.52mmol) and 4-dimethylaminopyridine (45mg, 0.37mmol) were added to the system, Add 4-bromo-3-fluorobenzenesulfonyl chloride (500mg, 1.84mmol) at 0°C 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-pyridine Azol-3-yl) cyclopentyl isopropyl carbamate 23c

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

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)cyclopentyl Isopropyl carbamate 23

Compound 23 was prepared according to the synthesis steps of Example 11 by replacing the corresponding raw materials. (19 mg, 38.2% yield). MS m/z (ESI): 443.2 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d6): δ 11.99(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)cyclopentylisopropylamine carbamate

Compound 24 (19 mg, yield 38.2%) was obtained by replacing the corresponding raw materials according to the synthetic steps of Example 23 . MS m/z (ESI): 425.2 [M+H] ⁺ .

¹ H NMR (400MHz,DMSO- d ₆ ): δ 11.91(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)ring Amyl isopropyl carbamate

first step

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

At room temperature, dissolve deuterated methylamine hydrochloride (210mg, 3mmol) and DIEA (650mg, 5mmol) in anhydrous dichloromethane (10mL), and slowly add 5-bromopyridine-2-sulfonyl chloride after clarification (250mg, 1mmol), continue to stir, TLC monitors until the reaction is over, add water (10mL) to wash once, and saturated brine (10mL) to wash once, collect the organic phase layer anhydrous sodium sulfate and dry, 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 -pyrazol-3-yl)cyclopentylisopropylcarbamate 25c

Under nitrogen atmosphere, 5-bromo-N-(methyl-d3)pyridine-2-sulfonamide (25.3 mg, 0.10 mmol), (1R,3S)-3-(5-amino-1-( 3rd-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" ), Pd2(dba)3 (9.1mg, 0.01mmol), Xantphos (11.4mg, 0.02mmol), CS2CO3 (96.2mg, 0.30mmol) were dissolved in 1,4-dioxane (2mL), microwave at 80°C The reaction was carried out for 2 hours. The reaction solution was cooled to room temperature, filtered, and the filtrate was collected. After concentration under reduced pressure, normal phase separation gave 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)ring Amyl isopropyl 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)-Second butyl)urethane

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

MS m/z (ESI): 437.2 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO- d ₆ ): δ 12.01(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)cyclopentylisopropyl base urethane

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

MS m/z (ESI): 423.2 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO- d ₆ ): δ 12.03(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 synthesis 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 ): δ 12.08(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 ): δ 11.97(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 ): δ 11.95(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)cyclopentylisopropyl Urethane

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 , TLC monitoring to the end of the reaction, adding water (10mL) to wash once, saturated brine (10mL) to wash 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, 74% yield).

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 after the reaction is over, filter the precipitate, add water (10mL) to wash once, and saturated brine (10mL) to wash once, collect the organic phase 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-pyrazole-3- base) cyclopentyl isopropyl carbamate 31d

Under nitrogen atmosphere, 2-chloro-5-((methylsulfonyl)methyl)pyridine (20.6 mg, 0.10 mmol), (1R,3S)-3-(5-amino-1-(third -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), microwave 80 ℃ for 2 hours. The reaction solution was cooled to room temperature, filtered, and the filtrate was collected, concentrated under reduced pressure and separated in 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)cyclopentylisopropyl Urethane 31

(1R,3S)-3-(1-(tert-butyl)-5-((5-((methylsulfonyl)methyl)pyridin-2-yl)amino)-1H-pyrazole-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 synthetic 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 ): δ 11.81(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-thio]phenyl }amino)-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 ): δ 11.99(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 =6.8,2.1Hz,6H).

Example 34

(Propan-2-ylamino)methanoic acid-(1R,3S)-3-{5-[(4-{[(2-hydroxyethyl)amino]dioxylidene-6-thio}benzene Base)amino]-1H-pyrazol-3-yl}cyclopentyl ester

Compound 34 was prepared according to the synthetic 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}cyclopenta base 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 ): δ 12.21(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 }cyclopentyl ester

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 ): δ 12.12(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-yl)cyclopentyl ester

Compound 37 was prepared according to the synthesis 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)methanoic acid-(1R,3S)-3-[5-({4-[dioxylidene(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 the 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}cyclopenta base 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-(methyldioxylidene-6-thio)hexahydropyridin-4-yl]amino }-2H-pyrazol-3-yl)cyclopentyl ester

first step

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

Compound 1-N-methylsulfonyl-4-piperidone (178mg, 1.0mmol), 1a (310mg, 1.0mmol), was dissolved in dichloromethane (10mL), 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-(methyldioxylidene-6-thio)hexahydropyridin-4-yl]amino }-2H-pyrazol-3-yl)cyclopentyl ester 40

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

MS m/z (ESI): 414.3 [M+H] ⁺ .

Example 41

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

Compound 41 was prepared according to the synthesis 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 ): δ 11.94(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-pyrazole-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 Disclosed on Ovarian Cancer Cells (OVCAR3)

Table 1. Experimental materials and instruments

Experimental procedure

Ovarian cancer cells OVCAR3 were cultured in RPMI 1640 with 10% FBS in a 37% incubator with 5% CO ₂ . 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 10uM, 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.

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

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

400nM，400nM<B

600nM，600nM<C

1000nM。 The IC ₅₀ (nM) of the disclosed compound on OVCAR3, A

400nM, 400nM<B

600nM, 600nM<C

1000nM.

Table 2. The IC ₅₀ (nM) of the disclosed compounds against OVCAR3

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 1000 g for 1 min; prepare 2x enzyme in 1x kinase buffer (40 mM Tris-HCl, pH 7.4, 20 mM MgCl2, 0.1 mg/ml BSA, 50 uM DTT) and mix 2.5 μl of 2x enzyme A 384-well assay plate was added, and the plate was centrifuged at 1000g for 30s and left at room temperature for 10 minutes. Prepare 2x substrate and ATP mix in 1x kinase buffer and start the reaction by adding 2.5 μl of 2x substrate and ATP mix. The plate was centrifuged at 1000g for 30 seconds, the assay plate was sealed, and reacted at room temperature for 1 hour. Add 4 μl of ADP-Glo reagent and incubate at room temperature for 40 minutes, then add 8 μl of kinase detection reagent and incubate at room temperature for 40 minutes.

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: average signal of positive control; Ave_VC signal: average signal of vehicle group.

_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 the disclosed compounds against CDK1/B, CDK2/Cyclin E, CDK9/Cyclin T1

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 drug concentration in the blood plasma at different times after oral administration of Example 8 and Example 9 was determined by LC/MS/MS method. To study the pharmacokinetic behavior of the compound of the present invention in the body of Balb/c nude mouse, and to 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 the 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, the R ^and R are ^each independently selected from hydrogen, alkyl, haloalkyl, hydroxyalkyl, cycloalkyl or heterocycloalkyl, and the alkyl, haloalkyl, hydroxyalkyl, cycloalkyl Or heterocycloalkyl is optionally substituted by one or more substituents independently selected from R ^1A ; The R ^1A is selected from halogen, hydroxy, alkyl, alkoxy, haloalkyl, haloalkoxy, heterocycloalkyl or NR'(R"), the alkyl, alkoxy, haloalkyl, halo Alkoxy or heterocycloalkyl is optionally substituted by one or more substituents independently selected from R ⁵ ; Or the 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 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 R ^{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 pendant oxy, 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 with ring A are optionally substituted by one or more independently selected from R ^4A

or

; The R ^4A is selected from halogen, cyano, nitro, hydroxy, 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 optionally substituted by one or more substituents independently selected from R ⁸ ; The X is independently selected from O or S; The R ^is selected from halogen, hydroxy, 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; The n is selected from 1, 2, 3, 4 or 5. The compound or a pharmaceutically acceptable salt thereof as claimed in claim 1, which 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 as defined in Claim 1 respectively. The compound or pharmaceutically acceptable salt thereof as described in Claim 1, which is a compound represented by formula III or IV or a pharmaceutically acceptable salt thereof,

Wherein, the R ¹ , R ² , R ⁴ and n are as defined in Claim 1 respectively. A compound or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 3, wherein ring A is selected from phenyl, five-membered nitrogen-containing heteroaryl or six-membered nitrogen-containing heteroaryl; preferably pyridine Base, pyrimidinyl, piperidinyl, imidazolyl, pyrrolyl or pyrazolyl; most preferably pyridinyl. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 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 , G2, 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 or pharmaceutically acceptable salt thereof as claimed in item 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 R ⁴ is as defined in Claim 1. A compound or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 4, wherein ^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 as described in any one of claims 1 to 5, wherein, the ^R is selected from alkyl, haloalkyl, hydroxyalkyl, cycloalkyl or heterocycloalkyl, the Alkyl, haloalkyl, hydroxyalkyl, cycloalkyl or heterocycloalkyl is optionally substituted by one or more substituents independently selected from ^{R 1A selected} from haloalkyl, alkoxy, Haloalkyl, haloalkoxy, heterocycloalkyl or NR'(R"), the alkyl, alkoxy, haloalkyl, haloalkoxy, heterocycloalkyl optionally replaced by one or more R ⁵ replaced; Preferably, the 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 , the R ^1A is selected from halogen or C _1-6 alkyl; More preferably, the ^R is selected from methyl, ethyl, isopropyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, and the cyclopropyl, cyclobutyl, cyclopentyl are replaced by one Or a plurality of R ^1A substitution, the R ^1A is selected from methyl, ethyl, isopropyl; Particularly preferably, the R ² is selected from isopropyl or methylcyclopropyl. The compound or pharmaceutically acceptable salt thereof as described in any one of claims 1 to 2, 4 to 8, wherein, the ^R is selected from

; Wherein X is O, said R ⁶ and R ⁷ are each independently selected from hydrogen, C _1-6 alkyl, preferably hydrogen, methyl, ethyl. The compound or pharmaceutically acceptable salt thereof as claimed in any one of items 1 to 9, wherein each ^R is independently selected from hydrogen, cyano, halogen, hydroxyl, alkyl, alkoxy, haloalkane radical, 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 ; The 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

, The R' and R" are as defined in Claim 1 respectively. A compound or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 9, wherein each R ⁴ is independently selected from hydrogen, cyano or halogen; Alternatively, each 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 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; The R' and R" are as defined in Claim 1 respectively. The compound according to any one of claims 1 to 11 or a pharmaceutically acceptable salt thereof, wherein the R ¹ is hydrogen, and the R ² is selected from isopropyl or methylcyclopropyl. The compound or pharmaceutically acceptable salt thereof as described in any one of claims 1 to 12, wherein, the R' and R" are each independently selected from hydrogen or C _1-6 alkyl, preferably hydrogen or methyl , Ethyl. The compound or a pharmaceutically acceptable salt thereof as claimed in any one of claims 3 to 8, 10 to 13, wherein 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, each R ⁴ is 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; The n is selected from 1, 2, 3, 4 or 5. The compound or pharmaceutically acceptable salt thereof as described in any one of claims 3 to 14, wherein, the

selected from

,

or

, better

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

An isotope substitution of the compound as described in 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 described in any one of Claims 1 to 16, the isotope substitution as described in Claim 17, and a pharmaceutically acceptable excipient. A compound as described in any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof, an isotope substitution as described in claim 17, or a pharmaceutical composition as described in claim 18 is used in the preparation of /or use in medicines for treating diseases related to protein-dependent kinases, the protein-dependent kinases Preferably CDK2, the disease related to protein-dependent kinase is preferably cell proliferative disease, cancer or immune disease. A compound as described in any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof, an isotope substitution as described in claim 17, or a pharmaceutical composition as described in claim 18 is used in the preparation of /or use in medicines for treating cyclin-related diseases, the cyclin is preferably cyclin E, more preferably cyclin E1, and cyclin E2, and the cyclin-related disease is preferably cell proliferation Disease, cancer or immune disease. A compound as described in any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof, an isotope substitution as described in claim 17, or a pharmaceutical composition as described in claim 18 is used in the preparation of /or use in a medicament for the treatment of cancer 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, or sarcoma. A preparation method of the compound or its pharmaceutically acceptable salt as described in any one of claim items 3 to 9, 11 to 16, or the isotope substitution as described in claim item 17, which comprises the compound shown in formula V in acidic The step of removing the amino protecting group PG under 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 tertiary butyl, acetyl, trifluoroacetyl, and trityl , benzyl, formyl, the best is tertiary 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 tertiary butyl, acetyl, trifluoroacetyl, and trityl , benzyl, formyl, the best is tertiary butyl.