CN112142763B

CN112142763B - Pyrazolopyrimidine compound, and preparation method and application thereof

Info

Publication number: CN112142763B
Application number: CN202010600148.4A
Authority: CN
Inventors: 王倩; 张霖; 夏广新; 楼江松; 葛辉; 霍国永; 舒思杰; 石辰; 张弛; 张智慧; 毛煜; 张冰宾; 余建鑫; 柯樱; 刘彦君
Original assignee: Shanghai Pharmaceuticals Holding Co Ltd
Current assignee: Shanghai Pharmaceuticals Holding Co Ltd
Priority date: 2019-06-28
Filing date: 2020-06-28
Publication date: 2024-01-26
Anticipated expiration: 2040-06-28
Also published as: CN112142763A

Abstract

The invention discloses pyrazolopyrimidine compound, a preparation method and application thereof. The invention provides pyrazolopyrimidine compound shown as a formula A, which has better inhibitory activity on WEE1 kinase.

Description

Pyrazolopyrimidine compound, and preparation method and application thereof

Technical Field

The invention relates to pyrazolopyrimidine compound, a preparation method and application thereof.

Background

Cell cycle is closely related to the DNA damage repair process. The cell cycle refers to the whole process undergone by cell division, and is divided into two phases, namely an interphase (interphase) and a mitotic phase (M). Cell cycle checkpoint (checkpoint) is a key point in regulating the cell cycle, and is mainly used to ensure that each event in the cycle can be completed in time and order, and that the cell state is regulated to adapt to the external environment.

The main checkpoints of the cells are: 1) G1/S checkpoint: in mammals, designated as point R (restriction), controls the entry of cells from quiescent G1 phase into DNA synthesis phase; 2) S-phase test point: whether the DNA replication is complete; 3) G2/M checkpoint: is a control point that regulates the entry of cells into the dividing phase; 4) Mid-late checkpoint: also known as spindle assembly checkpoint, can cause disruption of the cell cycle if the centromere is not properly attached to the spindle. If there is an abnormality in some of the processes of the cell division cycle, such as DNA damage, the checkpoint will sense and initiate repair in time. The P53 protein is an important protein for regulating the G1 test point, and when DNA is damaged, the protein prevents cells from entering the S phase, activates a DNA repair mechanism and is important for maintaining the integrity of the genome of the cells. However, since tumor cells often have P53 mutations that cause defects in the G1 checkpoint, regulation of the cell division cycle in P53 mutated cells is dependent on the G2/M checkpoint.

WEE1 kinase is a cell cycle regulating protein, can regulate the phosphorylation state of cyclin-dependent kinase 1 (CDK 1), thus regulating the activity of CDK1 and cyclin B (cyclin B) complex, thus realizing the regulation of cell cycle, and has important regulation function on DNA damage check points. WEE1 is a key gene for G2/M phase retardation, plays an important role in monitoring, and over-expression, inhibition or down-regulation of WEE1 kinase in some cancers can trigger mitosis disasters, so WEE1 kinase inhibitors have a key role in anticancer treatment and are now hot spots for developing anticancer agents.

International patent applications WO2019037678, WO2019028008, WO2018133829, WO2010098367, WO2010067886, WO2008115742, WO2008115738, WO2007126122, WO2007126128, WO2004007499 and the like disclose partial small molecule WEE1 kinase inhibitors, but no small molecule WEE1 kinase inhibitors are currently marketed, and there is a need in the art to develop novel WEE1 kinase inhibitors with good anticancer activity and high safety.

Disclosure of Invention

The invention aims to solve the technical problem that the existing compound with the inhibitory activity to WEE1 kinase has a single structure, so the invention provides a pyrazolopyrimidine compound, a preparation method and application thereof, and the compound has better inhibitory activity to WEE1 kinase.

The invention provides pyrazolopyrimidine compound shown as a formula A or pharmaceutically acceptable salt thereof;

x is a single bond or CH ₂ ；

Y is N or CH;

z is a single bond or CH ₂ ；

R ¹ Is H, -CN, -C (=O) R ^1-1 、-C(＝NR ^1-2 )R ^1-3 、-S(＝O) ₂ R ^1-4 、Or, optionally, by 1, 2 or 3R ^1-5 Substituted: c (C) ₁ ～C ₇ Alkyl, C ₂ ～C ₇ Alkenyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl, C ₆ ～C ₁₀ Aryl or C ₁ ～C ₇ Heteroaryl;

R ^1-1 is hydrogen, cyano, -NR ^1-1-1 R ^1-1-2 、-OR ^1-1-3 Or, optionally, by 1, 2 or 3R ^1-1-4 Substituted: c (C) ₁ ～C ₇ Alkyl, C ₂ ～C ₇ Alkenyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl, C ₆ ～C ₁₀ Aryl or C ₁ ～C ₇ Heteroaryl;

R ^1-1-1 and R is ^1-1-2 Independently hydrogen, C ₁ ～C ₇ Alkyl, C ₂ ～C ₇ Alkenyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl, C ₆ ～C ₁₀ Aryl or C ₁ ～C ₇ Heteroaryl;

alternatively, R ^1-1-1 、R ^1-1-2 And the nitrogen atom to which it is attached forms C ₃ ～C ₁₄ A heterocycloalkyl group; the C is ₃ ～C ₁₄ 1 or more than 2 methylene groups in the heterocycloalkyl group are optionally and independently replaced by oxygen atoms,Sulfur atom, sulfinyl, sulfonyl, carbonyl, vinylidene or-N (R) ^1-1-1-1 ) -the indicated group substitution; r is R ^1-1-1-1 Independently halogen, -OH, C ₁ ～C ₇ Alkyl or C ₃ ～C ₁₄ Cycloalkyl;

R ^1-1-3 is hydrogen, C ₁ ～C ₇ Alkyl, C ₂ ～C ₇ Alkenyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl, C ₆ ～C ₁₀ Aryl or C ₁ ～C ₇ Heteroaryl;

R ^1-1-4 independently halogen, -OH, -CN, amino, mercapto, C ₁ ～C ₇ Alkyl, C ₁ ～C ₇ Alkoxy, C ₁ ～C ₇ Alkylthio, C ₂ ～C ₇ Alkenyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl, C ₆ ～C ₁₀ Aryl, C ₁ ～C ₇ Heteroaryl, or, substituted with 1 or 2R ^1-1-4-1 Substituted amino groups; r is R ^1-1-4-1 Independently C ₁ ～C ₇ Alkyl or C ₃ ～C ₁₄ Cycloalkyl;

R ^1-2 is hydrogen, -CN, -OR ^1-2-1 Or C ₁ ～C ₇ An alkyl group;

R ^1-2-1 is hydrogen, C ₁ ～C ₇ Alkyl, C ₂ ～C ₇ Alkenyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl, C ₆ ～C ₁₀ Aryl or C ₁ ～C ₇ Heteroaryl;

R ^1-3 is-NR ^1-3-1 R ^1-3-2 Or, optionally, by 1, 2 or 3R ^1-3-3 Substituted: c (C) ₁ ～C ₇ Alkyl, C ₂ ～C ₇ Alkenyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl, C ₆ ～C ₁₀ Aryl or C ₁ ～C ₇ Heteroaryl;

R ^1-3-1 and R is ^1-3-2 Independently hydrogen, C ₁ ～C ₇ Alkyl, C ₂ ～C ₇ Alkenyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl, C ₆ ～C ₁₀ Aryl or C ₁ ～C ₇ Heteroaryl;

alternatively, R ^1-3-1 、R ^1-3-2 And the nitrogen atom to which it is attached forms C ₃ ～C ₁₄ A heterocycloalkyl group; the C is ₃ ～C ₁₄ 1 or more methylene groups in the heterocycloalkyl group are optionally, independently substituted by oxygen, sulfur, sulfinyl, sulfonyl, carbonyl, vinylidene or-N (R) ^1-3-1-1 ) -the indicated group substitution; r is R ^1-3-1-1 Independently halogen, -OH, C ₁ ～C ₇ Alkyl or C ₃ ～C ₁₄ Cycloalkyl;

R ^1-3-3 independently halogen, -OH, -CN, amino, mercapto, C ₁ ～C ₇ Alkyl, C ₁ ～C ₇ Alkoxy, C ₁ ～C ₇ Alkylthio, C ₂ ～C ₇ Alkenyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl, C ₆ ～C ₁₀ Aryl, C ₁ ～C ₇ Heteroaryl, or, substituted with 1 or 2R ^1-3-3-1 Substituted amino groups; r is R ^1-3-3-1 Independently C ₁ ～C ₇ Alkyl or C ₃ ～C ₁₄ Cycloalkyl;

R ^1-4 is-OH, -NR ^1-4-1 R ^1-4-2 Or, optionally, by 1, 2 or 3R ^1-4-3 Substituted: c (C) ₁ ～C ₇ Alkyl, C ₂ ～C ₇ Alkenyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl, C ₆ ～C ₁₀ Aryl, or, C ₁ ～C ₇ Heteroaryl;

R ^1-4-1 and R is ^1-4-2 Independently hydrogen, C ₁ ～C ₇ Alkyl, C ₂ ～C ₇ Alkenyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl, C ₆ ～C ₁₀ Aryl or C ₁ ～C ₇ Heteroaryl;

alternatively, R ^1-4-1 、R ^1-4-2 And the nitrogen atom to which it is attached forms C ₃ ～C ₁₄ A heterocycloalkyl group; the C is ₃ ～C ₁₄ 1 or more methylene groups in the heterocycloalkyl group are optionally, independently substituted by oxygen, sulfur, sulfinyl, sulfonyl, carbonyl, vinylidene or-N (R) ^1-4-1-1 ) -the indicated group substitution; r is R ^1-4-1-1 Independently halogen, -OH, C ₁ ～C ₇ Alkyl or C ₃ ～C ₁₄ Cycloalkyl;

R ^1-4-3 independently halogen, -OH, -CN, amino, mercapto, C ₁ ～C ₇ Alkyl, C ₁ ～C ₇ Alkoxy, C ₁ ～C ₇ Alkylthio, C ₂ ～C ₇ Alkenyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl, C ₆ ～C ₁₀ Aryl, C ₁ ～C ₇ Heteroaryl, or, substituted with 1 or 2R ^1-4-3-1 Substituted amino groups; r is R ^1-4-3-1 Independently C ₁ ～C ₇ Alkyl or C ₃ ～C ₁₄ Cycloalkyl;

R ^1-5 independently halogen, -OH, -SH, -CN, -NR ^1-5-1 R ^1-5-2 、-(C＝O)R ^1-5-3 Or, optionally, by 1, 2 or 3R ^1-5-4 Substituted: c (C) ₁ ～C ₇ Alkyl, C ₂ ～C ₇ Alkenyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl, C ₁ ～C ₇ Alkoxy, C ₁ ～C ₇ Alkylthio, C ₃ ～C ₁₄ Heterocycloalkyl, C ₆ ～C ₁₀ Aryl or C ₁ ～C ₁₀ Heteroaryl;

R ^1-5-1 and R is ^1-5-2 Independently hydrogen, C ₁ ～C ₇ Alkyl, C ₂ ～C ₇ Alkenyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl, C ₆ ～C ₁₀ Aryl or C ₁ ～C ₇ Heteroaryl;

alternatively, R ^1-5-1 、R ^1-5-2 And the nitrogen atom to which it is attached forms C ₃ ～C ₁₄ A heterocycloalkyl group; the C is ₃ ～C ₁₄ 1 or more methylene groups in the heterocycloalkyl group are optionally, independently substituted by oxygen, sulfur, sulfinyl, sulfonyl, carbonyl, vinylidene or-N (R) ^1-5-1-1 ) -the indicated group substitution; r is R ^1-5-1-1 Independently halogen, -OH, C ₁ ～C ₇ Alkyl or C ₃ ～C ₁₄ Cycloalkyl;

R ^1-5-3 independently hydrogen, -OR ^1-5-3-1 、-NR ^1-5-3-2 R ^1-5-3-3 、C ₁ ～C ₇ Alkyl, C ₂ ～C ₇ Alkenyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl, C ₆ ～C ₁₀ Aryl or C ₁ ～C ₇ Heteroaryl;

R ^1-5-3-1 、R ^1-5-3-2 and R is ^1-5-3-3 Independently hydrogen, C ₁ ～C ₇ Alkyl, C ₂ ～C ₇ Alkenyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl, C ₆ ～C ₁₀ Aryl or C ₁ ～C ₇ Heteroaryl;

R ^1-5-4 independently halogen, -OH, -CN, amino, mercapto, C ₁ ～C ₇ Alkyl, C ₁ ～C ₇ Alkoxy, C ₁ ～C ₇ Alkylthio, C ₂ ～C ₇ Alkenyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl, C ₆ ～C ₁₀ Aryl, C ₁ ～C ₇ Heteroaryl, or, substituted with 1 or 2R ^1-5-4-1 Substituted amino groups; r is R ^1-5-4-1 Independently C ₁ ～C ₇ Alkyl or C ₃ ～C ₁₄ Cycloalkyl;

R ^1-6 and R is ^1-7 Independently hydrogen or C ₁ ～C ₇ An alkyl group;

R ² optionally by 1, 2 or 3R ^2-1 Substituted: c (C) ₁ ～C ₇ Alkyl, C ₃ ～C ₁₄ Cycloalkyl or C ₃ ～C ₁₄ A heterocycloalkyl group;

R ^2-1 independently halogen, hydroxy, amino, C ₁ ～C ₇ Alkyl, C ₁ ～C ₇ Alkoxy, C ₃ ～C ₁₄ Cycloalkyl or C ₃ ～C ₁₄ A heterocycloalkyl group;

in any of the above cases, the above C ₃ ～C ₁₄ Heterocycloalkyl, C ₁ ～C ₇ The heteroatoms in the heteroaryl group are independently selected from one or more of boron, silicon, oxygen, sulfur, selenium, nitrogen, and phosphorus; the number of heteroatoms is independently 1, 2, 3 or 4.

In one embodiment, certain substituents in the pyrazolopyrimidine compound of formula a or a pharmaceutically acceptable salt thereof may further have the following definitions, and the definitions of substituents not referred to below are as described in any of the above embodiments (hereinafter referred to simply as "in one embodiment"):

the pyrazolopyrimidine compound shown in the formula A is not

In one embodiment:

the said processIs->

In one aspect of the present invention,for example-> Also for example->Also for example->

In a certain scheme, the pyrazolopyrimidine compound shown in the formula A is a compound shown in the formula I

The said processIs->

In one embodiment:

when R is ¹ Optionally by 1, 2 or 3R ^1-5 Substituted C ₁ ～C ₇ In the case of alkyl, said C ₁ ～C ₇ Alkyl radicals such as C ₁ ～C ₃ Alkyl, such as methyl, ethyl, n-propyl or isopropyl.

In one embodiment:

when R is ^1-5 Independently a halogen, such as fluorine, chlorine, bromine or iodine, and such as fluorine.

In one embodiment:

when R is ¹ Is 1, 2 or 3R ^1-5 Substituted C ₁ ～C ₇ Alkyl, R ^1-5 Independently halogen, said "is substituted with 1, 2 or 3R ^1-5 Substituted C ₁ ～C ₇ The alkyl "may be 2, 2-difluoroethyl or 3, 3-trifluoropropyl.

In one embodiment:

when R is ¹ Is 1R ^1-5 Substituted C ₁ ～C ₇ Alkyl, R ^1-5 When independently CN, said "is substituted with 1R ^1-5 Substituted C ₁ ～C ₇ The alkyl "may be 2-cyanoethyl.

In one embodiment:

when R is ^1-5 Independently C ₃ ～C ₁₄ In the case of cycloalkyl, said C ₃ ～C ₁₄ Cycloalkyl radicals such as C ₃ ～C ₁₄ Monocyclic cycloalkyl, C ₃ ～C ₁₄ Spiro cycloalkyl, C ₃ ～C ₁₄ Condensed ring cycloalkyl or C ₃ ～C ₁₄ Bridged cycloalkyl groups.

The C is ₃ ～C ₁₄ Monocyclic cycloalkyl groups such as C ₃ ～C ₆ Monocyclic cycloalkyl, such as again cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, and also such as cyclobutyl, cyclopentyl or cyclohexyl.

The C is ₃ ～C ₁₄ Bridged cycloalkyl radicals such as C ₅ ～C ₈ Bridged cycloalkyl radicals, again e.gOr->

In one embodiment:

when R is ^1-5 Independently C ₃ ～C ₁₄ In the case of cycloalkyl, said C ₃ ～C ₁₄ Cycloalkyl radicals such as C ₃ ～C ₁₄ Saturated cycloalkyl groups.

In one embodiment:

When R is ¹ Is 1R ^1-5 Substituted C ₁ ～C ₇ Alkyl, R ^1-5 Independently C ₃ ～C ₁₄ In the case of cycloalkyl, the term "is substituted with 1R ^1-5 Substituted C ₁ ～C ₇ The alkyl "may be cyclopropylmethyl or cyclohexylmethyl.

In one embodiment:

when R is ^1-5-3-1 Is C ₁ ～C ₇ In the case of alkyl, said C ₁ ～C ₇ Alkyl radicals such as C ₁ ～C ₃ Alkyl, such as methyl, ethyl, n-propyl or isopropyl, and also ethyl.

In one embodiment:

when R is ¹ Optionally by 1, 2 or 3R ^1-5 Substituted C ₃ ～C ₁₄ In the case of cycloalkyl, said C ₃ ～C ₁₄ Cycloalkyl radicals such as C ₃ ～C ₁₄ Monocyclic cycloalkyl, C ₃ ～C ₁₄ Spiro cycloalkyl, C ₃ ～C ₁₄ Condensed ring cycloalkyl or C ₃ ～C ₁₄ Bridged cycloalkyl groups.

In one embodiment:

when R is ¹ Optionally by 1, 2 or 3R ^1-5 Substituted C ₃ ～C ₁₄ In the case of cycloalkyl, said C ₃ ～C ₁₄ Cycloalkyl radicals such as C ₃ ～C ₁₄ Saturated cycloalkyl groups.

In one embodiment:

when R is ¹ Is 1R ^1-5 Substituted C ₃ ～C ₁₄ Cycloalkyl, R ^1-5 Is independently-NR ^1-5-1 R ^1-5-2 When said is 1R ^1-5 Substituted C ₃ ～C ₁₄ Cycloalkyl "is, for example, 3-aminocyclobutyl.

In one embodiment:

When R is ¹ Optionally by 1, 2 or 3R ^1-5 Substituted C ₃ ～C ₁₄ In the case of heterocycloalkyl, said C ₃ ～C ₁₄ Heterocycloalkyl radicals such as C ₃ ～C ₁₄ Monocyclic heterocycloalkyl, C ₃ ～C ₁₄ Spirocyclic heterocycloalkyl, C ₃ ～C ₁₄ Condensed ring heterocycloalkyl or C ₃ ～C ₁₄ Bridged heterocycloalkyl.

The C is ₃ ～C ₁₄ Monocyclic heterocycloalkyl, for example, "C having 1 or 2 hetero atoms selected from one or both of N, O and S ₃ ～C ₉ Monocyclic heterocycloalkyl ", for example also" C having 1 or 2 heteroatoms selected from one or both of N, O and S ₃ ～C ₅ Monocyclic heterocycloalkyl ", also for example" C having 1 or 2 hetero atoms selected from one or both of N, O and S ₃ ～C ₅ Monocyclic heterocycloalkyl ", and which is attached to the benzene ring through a carbon atom, such as oxetanyl or piperidinyl.

Piperidyl radicals such asSaid oxetanyl group being for example +.>

In one embodiment:

when R is ¹ Optionally by 1, 2 or 3R ^1-5 Substituted C ₃ ～C ₁₄ In the case of heterocycloalkyl, except R ^1-5 In addition, the C ₃ ～C ₁₄ The heteroatoms in the heterocycloalkyl group may be unsubstituted.

In one embodiment:

when R is ¹ Optionally by 1, 2 or 3R ^1-5 Substituted C ₃ ～C ₁₄ In the case of heterocycloalkyl, said C ₃ ～C ₁₄ The methylene group in the heterocycloalkyl group may be unsubstituted or substituted.

In one embodiment:

When R is ^1-5 Optionally by 1, 2 or 3R ^1-5-4 Substituted C ₁ ～C ₇ In the case of alkyl, said C ₁ ～C ₇ Alkyl radicals such as C ₁ ～C ₃ Alkyl, such as methyl, ethyl, n-propyl or isopropyl.

In one embodiment:

when R is ¹ Is optionally substituted with 1R ^1-5 Substituted C ₃ ～C ₁₄ Heterocycloalkyl, R ^1-5 Is C ₁ ～C ₇ In the case of alkyl radicals, the radicals "optionally substituted by 1R ^1-5 Substituted C ₃ ～C ₁₄ Heterocyclylalkyl radicals "such as

In one embodiment:

when R is ^1-1 Optionally by 1, 2 or 3R ^1-1-4 Substituted C ₁ ～C ₇ In the case of alkyl, said C ₁ ～C ₇ Alkyl radicals such as C ₁ ～C ₃ Alkyl, such as methyl, ethyl, n-propyl or isopropyl, and also such as methyl or ethyl.

In one embodiment:

when R is ^1-1 Is optionally substituted with 1R ^1-1-4 Substituted C ₁ ～C ₇ Alkyl, R ^1-1-4 In the case of-OH, the said "Optionally by 1R ^1-1-4 Substituted C ₁ ～C ₇ Alkyl "such as hydroxymethyl.

In one embodiment:

when R is ^1-1 Optionally by 1, 2 or 3R ^1-1-4 Substituted C ₃ ～C ₁₄ In the case of cycloalkyl, said C ₃ ～C ₁₄ Cycloalkyl radicals such as C ₃ ～C ₁₄ Monocyclic cycloalkyl, C ₃ ～C ₁₄ Spiro cycloalkyl, C ₃ ～C ₁₄ Condensed ring cycloalkyl or C ₃ ～C ₁₄ Bridged cycloalkyl groups.

The C is ₃ ～C ₁₄ Bridged cycloalkyl radicals such as C ₅ ～C ₈ Bridged cycloalkyl radicals, again e.g Or->

In one embodiment:

when R is ^1-1 Optionally by 1, 2 or 3R ^1-1-4 Substituted C ₃ ～C ₁₄ In the case of cycloalkyl, said C ₃ ～C ₁₄ Cycloalkyl radicals such as C ₃ ～C ₁₄ Saturated cycloalkyl groups.

In one embodiment:

when R is ^1-6 Is C ₁ ～C ₇ In the case of alkyl, said C ₁ ～C ₇ Alkyl groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, and also such as methyl or ethyl.

In one embodiment:

when R is ^1-7 Is C ₁ ～C ₇ In the case of alkyl, said C ₁ ～C ₇ Alkyl groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, and also such as methyl or ethyl.

In one embodiment:

when R is ² Optionally by 1, 2 or 3R ^2-1 Substituted C ₁ ～C ₇ In the case of alkyl, said C ₁ ～C ₇ Alkyl radicals such as C ₁ ～C ₄ Alkyl, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, and also such as isopropyl.

In one embodiment:

when R is ² Is 1R ^2-1 Substituted C ₁ ～C ₇ Alkyl, R ^2-1 In the case of hydroxyl, said quilt has 1R ^2-1 Substituted C ₁ ～C ₇ Alkyl groups such as

In one embodiment:

when R is ² Optionally by 1, 2 or 3R ^2-1 Substituted C ₃ ～C ₁₄ In the case of heterocycloalkyl, said C ₃ ～C ₁₄ Heterocycloalkyl radicals such as C ₃ ～C ₁₄ Monocyclic heterocycloalkyl, C ₃ ～C ₁₄ Spirocyclic heterocycloalkyl, C ₃ ～C ₁₄ Condensed ring heterocycloalkyl or C ₃ ～C ₁₄ Bridged heterocycloalkyl.

The C is ₃ ～C ₁₄ Monocyclic heterocycloalkyl, for example, "C having 1 or 2 hetero atoms selected from one or both of N, O and S ₃ ～C ₉ Monocyclic heterocycloalkyl ", for example also" C having 1 or 2 heteroatoms selected from one or both of N, O and S ₃ ～C ₅ Monocyclic heterocycloalkyl ", also for example" C having 1 or 2 hetero atoms selected from one or both of N, O and S ₃ ～C ₅ Monocyclic heterocycloalkyl ", and which is bound to the benzene ring through a carbon atom, e.g. oxetanyl, more e.g. oxetanylBut-3-yl.

In one embodiment:

when R is ² Optionally by 1, 2 or 3R ^2-1 Substituted C ₃ ～C ₁₄ In the case of heterocycloalkyl, except R ^2-1 In addition, the C ₃ ～C ₁₄ The heteroatoms in the heterocycloalkyl group may be unsubstituted.

In one embodiment:

when R is ² Optionally by 1, 2 or 3R ^2-1 Substituted C ₃ ～C ₁₄ In the case of heterocycloalkyl, said C ₃ ～C ₁₄ The methylene group in the heterocycloalkyl group may be unsubstituted or substituted.

In one embodiment:

when R is ² Is 1R ^2-1 Substituted C ₃ ～C ₁₄ Heterocycloalkyl, R ^2-1 In the case of halogen or hydroxy, said quilt has 1R ^2-1 Substituted C ₃ ～C ₁₄ Heterocycloalkyl radicals, e.g.

In one embodiment:

the atoms in the pyrazolopyrimidine compound shown in the formula A or the pharmaceutically acceptable salt thereof can exist in the form of natural abundance.

In one embodiment:

R ¹ is H, -C (=O) R ^1-1 、-C(＝NR ^1-2 )R ^1-3 、-S(＝O) ₂ R ^1-4 Or, optionally, by 1, 2 or 3R ^1-5 Substituted: c (C) ₁ ～C ₇ Alkyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl or C ₁ ～C ₁₀ Heteroaryl groups.

In one embodiment:

R ¹ is H, -C (=O) R ^1-1 、Or, optionally, is covered by1, 2 or 3R ^1-5 Substituted: c (C) ₁ ～C ₇ Alkyl, C ₃ ～C ₁₄ Cycloalkyl, or, C ₃ ～C ₁₄ A heterocycloalkyl group.

In one embodiment:

R ¹ is H, -C (=O) R ^1-1 Or, optionally, by 1, 2 or 3R ^1-5 Substituted: c (C) ₁ ～C ₇ Alkyl, C ₃ ～C ₁₄ Cycloalkyl, or, C ₃ ～C ₁₄ A heterocycloalkyl group.

In one embodiment:

R ^1-1 is-NR ^1-1-1 R ^1-1-2 、-OR ^1-1-3 Cyano, or, optionally, by 1R ^1-1-4 Substituted: c (C) ₁ ～C ₇ Alkyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl or C ₃ ～C ₁₄ A heterocycloalkyl group; r is R ^1-1-1 And R is ^1-1-2 Is hydrogen or C ₁ ～C ₇ An alkyl group; r is R ^1-1-3 Is hydrogen, C ₁ ～C ₇ Alkyl or C ₃ ～C ₁₄ Cycloalkyl; r is R ^1-1-4 Halogen, OH or C ₁ ～C ₇ An alkyl group.

In one embodiment:

R ^1-1 is optionally substituted with 1R ^1-1-4 Substituted: c (C) ₁ ～C ₇ Alkyl or C ₃ ～C ₁₄ Cycloalkyl; r is R ^1-1-4 is-OH.

In one embodiment:

R ^1-2 hydrogen, -CN or-OH.

In one embodiment:

R ^1-3 is hydrogen, -NR ^1-3-1 R ^1-3-2 Or C ₃ ～C ₁₄ A heterocycloalkyl group; r is R ^1-3-1 And R is ^1-3-2 Independently hydrogen or C ₁ ～C ₇ An alkyl group.

In one embodiment:

R ^1-5 independently halogen, -CN, -NR ^1-5-1 R ^1-5-2 、-(C＝O)R ^1-5-3 、C ₁ ～C ₇ Alkyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl or C ₁ ～C ₁₀ Heteroaryl;

R ^1-5-1 and R is ^1-5-2 Independently hydrogen or C ₁ ～C ₇ An alkyl group;

R ^1-5-3 independently hydrogen, -OR ^1-5-3-1 Or C ₁ ～C ₇ An alkyl group; r is R ^1-5-3-1 Is C ₁ ～C ₇ An alkyl group.

In one embodiment:

R ^1-5 independently halogen, -CN, -NR ^1-5-1 R ^1-5-2 、-(C＝O)R ^1-5-3 、C ₁ ～C ₇ Alkyl or C ₃ ～C ₁₄ Cycloalkyl;

R ^1-5-1 and R is ^1-5-2 Is hydrogen;

R ^1-5-3 independently is-OR ^1-5-3-1 ；R ^1-5-3-1 Independently C ₁ ～C ₇ An alkyl group.

In one embodiment:

R ² is 1R ^2-1 Substituted: c (C) ₁ ～C ₇ Alkyl or C ₃ ～C ₁₄ A heterocycloalkyl group; r is R ^2-1 Independently a hydroxyl group or a halogen.

In one embodiment:

R ² is 1R ^2-1 Substituted: c (C) ₁ ～C ₇ Alkyl or C ₃ ～C ₁₄ A heterocycloalkyl group; r is R ^2-1 Is hydroxyl.

In one embodiment:

x is a single bond or CH ₂ The method comprises the steps of carrying out a first treatment on the surface of the Y is C; z is a single bond or CH ₂ 。

In one embodiment:

x is a single bond or CH ₂ The method comprises the steps of carrying out a first treatment on the surface of the Y is C; z is a single bond or CH ₂ ；

R ¹ Is that

R ^1-6 And R is ^1-7 Independently C ₁ ～C ₇ An alkyl group;

R ² optionally by 1, 2 or 3R ^2-1 Substituted: c (C) ₁ ～C ₇ Alkyl or C ₃ ～C ₁₄ A heterocycloalkyl group;

R ^2-1 is hydroxyl.

In one embodiment:

R ¹ is H, -C (=O) R ^1-1 、-C(＝NR ^1-2 )R ^1-3 、-S(＝O) ₂ R ^1-4 Or, optionally, by 1, 2 or 3R ^1-5 Substituted: c (C) ₁ ～C ₇ Alkyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl or C ₁ ～C ₁₀ Heteroaryl;

R ^1-1 is-NR ^1-1-1 R ^1-1-2 、-OR ^1-1-3 Cyano, or, optionally, by 1R ^1-1-4 Substituted: c (C) ₁ ～C ₇ Alkyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl or C ₃ ～C ₁₄ A heterocycloalkyl group; r is R ^1-1-1 And R is ^1-1-2 Is hydrogen or C ₁ ～C ₇ An alkyl group; r is R ^1-1-3 Is hydrogen, C ₁ ～C ₇ Alkyl or C ₃ ～C ₁₄ Cycloalkyl; r is R ^1-1-4 Halogen, OH or C ₁ ～C ₇ An alkyl group;

R ^1-2 hydrogen, -CN or-OH;

R ^1-3 is hydrogen, -NR ^1-3-1 R ^1-3-2 Or C ₃ ～C ₁₄ A heterocycloalkyl group; r is R ^1-3-1 And R is ^1-3-2 Independently hydrogen or C ₁ ～C ₇ An alkyl group;

R ^1-5-3 independently hydrogen, -OR ^1-5-3-1 Or C ₁ ～C ₇ An alkyl group; r is R ^1-5-3-1 Is C ₁ ～C ₇ An alkyl group;

In one embodiment:

R ¹ is H, -C (=O) R ^1-1 、Or, optionally, by 1, 2 or 3R ^1-5 Substituted: c (C) ₁ ～C ₇ Alkyl, C ₃ ～C ₁₄ Cycloalkyl, or, C ₃ ～C ₁₄ A heterocycloalkyl group;

R ^1-1 is optionally substituted with 1R ^1-1-4 Substituted: c (C) ₁ ～C ₇ Alkyl or C ₃ ～C ₁₄ Cycloalkyl; r is R ^1-1-4 is-OH;

R ^1-5-1 and R is ^1-5-2 Is hydrogen;

R ^1-5-3 independently is-OR ^1-5-3-1 ；R ^1-5-3-1 Independently C ₁ ～C ₇ An alkyl group;

R ^1-6 and R is ^1-7 Independently hydrogen or C ₁ ～C ₇ An alkyl group;

In one embodiment:

R ¹ is H, -C (=O) R ^1-1 Or, optionally, by 1, 2 or 3R ^1-5 Substituted: c (C) ₁ ～C ₇ Alkyl, C ₃ ～C ₁₄ Cycloalkyl, or, C ₃ ～C ₁₄ A heterocycloalkyl group;

R ^1-5-1 and R is ^1-5-2 Is hydrogen;

In one embodiment:

R ¹ is H, -C (=O) R ^1-1 、Or, optionally, by 1, 2 or 3R ^1-5 Substituted: c (C) ₁ ～C ₇ Alkyl, C ₃ ～C ₁₄ Cycloalkyl, or, C ₃ ～C ₁₄ A heterocycloalkyl group; />

R ^1-5-1 and R is ^1-5-2 Is hydrogen;

R ^1-6 and R is ^1-7 Independently hydrogen or C ₁ ～C ₇ An alkyl group;

In one embodiment:

R ^1-5-1 and R is ^1-5-2 Is hydrogen;

In a certain scheme, in the pyrazolopyrimidine compound shown in the formula A or the pharmaceutically acceptable salt thereof, the pyrazolopyrimidine compound shown in the formula A has any one of the following structures:

The invention also provides a preparation method of the compound A, which is any one of the following methods:

method 1: which comprises the following steps: step one, oxidizing a compound 1A in an organic solvent by using an oxidant to obtain a compound 1B; step two, reacting the compound 1B with a compound 1C 'under an organic solvent and alkaline conditions to obtain a compound 1D'; step three, removing the protecting group PG on the N of the compound 1D 'to obtain a compound 1E'; step four, carrying out reductive amination reaction on the compound 1E' and aldehyde or ketone in an organic solvent to obtain a compound A;

method 2: the compound 2A' and the compound 1A undergo substitution reaction to obtain a compound A;

method 3: compound 1E' and Compound R ^1-1 -COOH is subjected to condensation reaction to obtain a compound A;

the conditions and steps of the reaction described in method 1 may be those conventional in the art, with the following reaction conditions being particularly preferred in the present invention:

step one, the organic solvent is preferably one or more of methanol, dichloromethane, acetonitrile, toluene and DMF, more preferably dichloromethane or toluene; the oxidizing agent may be an oxidizing agent conventionally used in the art for oxidizing thioethers to sulfoxides, preferably m-chloroperoxybenzoic acid (m-CPBA).

Step two, the organic solvent is preferably dichloromethane or toluene; the basic conditions are preferably organic bases such as N, N-Diisopropylethylamine (DIPEA), triethylamine; the molar ratio of compound 1B, compound 1C', desipa is preferably 1:1:2, the reaction time is preferably 0 to 12 hours, and the reaction temperature is preferably 0 to 35 ℃.

Step three, the protecting group PG on N may be any suitable amino protecting group commonly used in the art, preferably Boc, with the aim of protecting compound 1C' from certain reactive groups (e.g. amino groups) on it when participating in the reaction; the conditions for removing the protecting group PG on N may be conventional conditions for removing various protecting groups in the art, such as hydrolysis conditions, aminolysis conditions, hydrogenation conditions, etc.; if Boc is used as the protecting group, the deprotection reaction may be carried out under standard conditions, e.g., methylene chloride/trifluoroacetic acid system, saturated hydrogen chloride dioxane solution; after the reaction for removing the protecting group is finished, the method can further comprise post-treatment operation; the methods and conditions of the post-treatment may be conventional in the art for such post-reaction treatments, preferably: washing, drying, filtering and evaporating the solvent from the reaction system, and then performing column chromatography; or evaporating the reaction system to remove the solvent, washing and filtering; or evaporating the reaction system to remove the solvent, and performing thin layer chromatography.

Step four, the solvent is methanol or dichloromethane preferentially; the reaction conditions for the reductive amination may be those commonly used in the art for reductive amination, preferably sodium borohydride acetate or sodium cyanoborohydride; the reaction time is preferably 0 to 12 hours; the reaction temperature is preferably 0℃to 35 ℃.

The conditions and steps of the reaction described in method 2 may be those conventional in the art, as described for the substitution reaction in method 1.

The conditions and steps of the reaction described in method 3 may be those conventional in the art.

The invention also provides a compound shown as a formula 1C 'or 2A':

wherein R is ¹ The definitions of X, Y, Z and PG are described above.

In one embodiment, the compound represented by formula 1C 'or 2A' may be any one of the following compounds:

the invention also provides a preparation method of the compound shown in the formula 1C', which comprises the following steps: step one, in a solvent, under the action of alkali, carrying out substitution reaction on a compound 1C1 'and halogenated nitrobenzene to obtain a compound 1C2'; step two, in a solvent, reducing a catalyst proto-body of the compound 1C2 'to obtain a compound 1C';

the conditions and steps of the reaction described in equation 1 may be conventional in the art, with the following reaction conditions being particularly preferred in the present invention:

in the first step, the solvent is preferably dimethylsulfoxide.

In step one, the base is preferably an inorganic base such as potassium carbonate, sodium carbonate, cesium carbonate, more preferably potassium carbonate.

In the first step, the halonitrobenzene is preferably fluoronitrobenzene or chloronitrobenzene, more preferably fluoronitrobenzene.

In the second step, the solvent is preferably methanol or ethanol.

In the second step, the reduction catalyst may be reduced iron powder, reduced zinc powder, selenious dichloride, platinum dioxide, palladium carbon, preferably palladium carbon; if palladium on carbon is used as the reduction catalyst, the hydrogen source employed may be a hydrogen source commonly used in the art, such as hydrogen, hydrazine hydrate, ammonium formate, preferably ammonium formate.

In the second step, the reaction temperature is preferably the solvent reflux temperature, and the reaction time is preferably 2-12 hours.

The invention also provides a preparation method of the compound shown in the formula 2A', which is any one of the following methods:

in method 1: step one, carrying out reductive amination reaction on a compound 1C1' and aldehyde or ketone to obtain a compound 2A1; step two, removing the protecting group PG on N in the compound 2A1 'to obtain a compound 2A2'; step three, carrying out substitution reaction on the compound 2A2 'and fluoronitrobenzene to obtain a compound 2A3'; step four, reducing the compound 2A3 'by a catalyst to obtain a compound 2A';

in method 2: step one, removing a protecting group PG on N in a compound 1C2 'to obtain a compound 2A4'; step two, carrying out substitution reaction on the compound 2A4 and a halogen substituted compound under alkaline conditions to obtain a compound 2A5', wherein the alkaline conditions are preferably potassium carbonate or silver oxide; the halogen substituted compound is preferably a bromide or an iodide; step three, reducing the compound 2A5 'by a catalyst to obtain a compound 2A';

The conditions and steps of the reaction described in equation 2 may be conventional in the art. The conditions and steps of the reaction described in equation 3 may be conventional in the art.

The invention also provides application of the substance X in preparing a kinase (such as WEE1 kinase) inhibitor;

the substance X is pyrazolopyrimidine compound shown in the formula A or pharmaceutically acceptable salt thereof.

The invention also provides application of the substance X in preparing medicines;

The invention also provides application of the substance X in preparing medicines; the medicine is used for treating and/or preventing diseases related to WEE1 kinase;

Such diseases associated with WEE1 kinase, e.g., cancer. Such as brain, head and neck, esophagus, thyroid, small cell, non-small cell, breast, lung, stomach, gall bladder-bile duct, liver, pancreas, colon, rectum, ovary, villous epithelium, uterus, cervix, renal pelvis-ureter, bladder, prostate, penis, testis, embryo, nephroblastoma, skin, malignant melanoma, neuroblastoma, osteosarcoma, ewing's tumor, soft tissue tumor, acute leukemia, chronic lymphoblastic leukemia, chronic myelogenous leukemia, or hodgkin's lymphoma, as well as breast, lung, pancreas, colon, ovary, acute leukemia, chronic lymphoblastic leukemia, chronic myelogenous leukemia, hodgkin's lymphoma, and colon or ovarian cancer.

The invention also provides application of the substance X in preparing medicines; the medicament is used for treating and/or preventing cancers;

Such as brain, head and neck, esophagus, thyroid, small cell, non-small cell, breast, lung, stomach, gall bladder-bile duct, liver, pancreas, colon, rectum, ovary, villous epithelium, uterus, cervix, renal pelvis-ureter, bladder, prostate, penis, testis, embryo, nephroblastoma, skin, malignant melanoma, neuroblastoma, osteosarcoma, ewing's tumor, soft tissue tumor, acute leukemia, chronic lymphoblastic leukemia, chronic myelogenous leukemia, or hodgkin's lymphoma, as well as breast, lung, pancreas, colon, ovary, acute leukemia, chronic lymphoblastic leukemia, chronic myelogenous leukemia, hodgkin's lymphoma, and colon or ovarian cancer.

The invention also provides a pharmaceutical composition comprising substance X and pharmaceutical adjuvant(s);

The present invention provides a combination comprising a substance X and an anticancer drug,

The anticancer drug may be any conventional anticancer drug in the art (but is not substance X described above), such as one or more of an anticancer alkylating agent, an anticancer metabolic antagonist, an anticancer antibiotic, a plant-derived anticancer agent, an anticancer platinum coordination compound, an anticancer camptothecin derivative, an anticancer tyrosine kinase inhibitor, a monoclonal antibody, an interferon, a biological response modifier, mitoxantrone, L-asparaginase, procarbazine, dacarbazine, hydroxyurea, prastatin, retinoic acid, alfasudite, albendazole Bei Ting, anastrozole, exemestane, bicalutamide, leuprorelin, flutamide, fulvestrant, pegaptanib sodium, desiccharum 2, aclacin, thyrotropina, arsenic trioxide, bortezomib, capecitabine and goserelin, and also such as an anticancer metabolic antagonist.

The anticancer alkylating agent may be one or more of the anticancer alkylating agents conventional in the art, such as nitrogen mustard N-oxide, cyclophosphamide, ifosfamide, milflan, busulfan, dibromomannitol, carboquinone, thiotepa, ramustine, nimustine, temozolomide and carmustine.

The anticancer metabolic antagonist may be one or more of the anticancer metabolic antagonists conventional in the art, such as methotrexate, 6-mercaptopurine nucleoside, mercaptopurine, 5-fluorouracil, tegafur, doxifluridine, carmofur, cytarabine sodium phosphate, enocitabine, S-1, gemcitabine, fludarabine, and pemetrexed disodium, and further such as 5-fluorouracil.

The anticancer antibiotic may be one or more of the anticancer antibiotics conventional in the art, such as actinomycin D, doxorubicin, daunorubicin, neocarcinomycin, bleomycin, pelomycin, mitomycin C, aclarubicin, pirarubicin, epirubicin, clobetadine Ding Sizhi, idarubicin, sirolimus and valrubicin.

The plant-derived anticancer agent may be a conventional plant-derived anticancer agent in the art, such as one or more of vincristine, vinblastine, vindesine, etoposide, solibuzocine, docetaxel, paclitaxel, and vinorelbine.

The anticancer platinum coordination compound may be one or more of the conventional anticancer platinum coordination compounds in the art, such as cisplatin, carboplatin, nedaplatin, and oxaliplatin.

The anticancer camptothecin derivative may be one or more of the anticancer camptothecin derivatives conventional in the art, such as irinotecan, topotecan and camptothecin.

The anti-cancer tyrosine kinase inhibitor may be an anti-cancer tyrosine kinase inhibitor conventional in the art, such as one or more of gefitinib, imatinib and erlotinib.

The monoclonal antibody may be a monoclonal antibody conventional in the art, such as one or more of cetuximab, bevacizumab, rituximab, alemtuzumab, and trastuzumab.

The interferon may be one or more of interferon conventional in the art, such as interferon alpha, interferon alpha-2 a, interferon alpha-2 b, interferon beta, interferon gamma-1 a and interferon gamma-n 1.

The biological response modifier can be a conventional biological response modifier in the art, such as one or more of coriolus versicolor polysaccharide, lentinan, siropyran, sartorine, and Wu Benmei.

The components of the combination may be used simultaneously or separately (e.g., sequentially); when the components of the combination are used simultaneously, the components of the combination may be mixed homogeneously (i.e., a mixture of the components).

The components of the combination may be formulated as a single pharmaceutical composition for simultaneous use, or the components may be formulated separately as separate pharmaceutical compositions (e.g., in kit form) for simultaneous use or separate use (e.g., for sequential use).

The invention also provides application of the combination in preparing a medicament for preventing and/or treating cancer.

In the use according to the invention, substance X described above, and the anticancer drug described above, can be administered simultaneously or separately (e.g. sequentially).

The invention also provides an application of the substance X in preparing medicines, wherein the medicines and the anticancer medicines are combined for preventing and/or treating cancers.

The invention also provides an application of the anti-cancer drug in preparing the drug, wherein the drug and the substance X are combined for preventing and/or treating cancers.

The invention also provides a pharmaceutical composition comprising the combination and (one or more) pharmaceutical excipients.

The pharmaceutical composition may consist of the combination and the pharmaceutical excipients.

The invention also provides a combined medicine box, which comprises a medicine composition A and a medicine composition B;

the medicine composition A comprises the substance X and (one or more) medicinal auxiliary materials;

the medicine composition B comprises the anticancer medicine and (one or more) medicinal auxiliary materials.

The combined kit can be composed of the medicine composition A and the medicine composition B.

The medicine composition A can be composed of a substance X and pharmaceutic adjuvant;

the medicine composition B can be composed of the anticancer medicine and pharmaceutic adjuvant.

The individual pharmaceutical compositions in the combination kit may be used simultaneously or separately (e.g., sequentially).

In a certain scheme, the pyrazolopyrimidine compound shown in the formula A or the pharmaceutically acceptable salt thereof can be pyrazolopyrimidine compound shown in the formula I or the pharmaceutically acceptable salt thereof;

R ¹ Is H, -CN, -C (=O) R ^1-1 、-C(＝NR ^1-2 )R ^1-3 、-S(＝O) ₂ R ^1-4 Or, optionally, by 1, 2 or 3R ^1-5 Substituted: c (C) ₁ ～C ₇ Alkyl, C ₂ ～C ₇ Alkenyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl, C ₆ ～C ₁₀ Aryl or C ₁ ～C ₇ Heteroaryl;

alternatively, R ^1-1-1 、R ^1-1-2 And the nitrogen atom to which it is attached forms C ₃ ～C ₁₄ A heterocycloalkyl group; the C is ₃ ～C ₁₄ 1 or more methylene groups in the heterocycloalkyl group are optionally, independently substituted by oxygen, sulfur, sulfinyl, sulfonyl, carbonyl, vinylidene or-N (R) ^1-1-1-1 ) -the indicated group substitution; r is R ^1-1-1-1 Independently halogen, -OH, C ₁ ～C ₇ Alkyl or C ₃ ～C ₁₄ Cycloalkyl;

R ^1-2 is hydrogen, -CN, -OR ^1-2-1 Or C ₁ ～C ₇ An alkyl group;

R ^1-3-1 and R is ^1-3-2 Independently hydrogen, C ₁ ～C ₇ Alkyl, C ₂ ～C ₇ Alkenyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl group、C ₆ ～C ₁₀ Aryl or C ₁ ～C ₇ Heteroaryl;

R ^1-5-4 independently halogen, -OH, -CN, amino, mercapto, C ₁ ～C ₇ Alkyl, C ₁ ～C ₇ Alkoxy, C ₁ ～C ₇ Alkylthio, C ₂ ～C ₇ Alkenyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl, C ₆ ～C ₁₀ Aryl, C ₁ ～C ₇ Heteroaryl, or, substituted with 1 or 2R ^1-5-4-1 Substituted amino groups; r is R ^1-5-4-1 Independently C ₁ ～C ₇ Alkyl or C ₃ ～C ₁₄ Cycloalkyl radicals；

in any of the above cases, the above C ₃ ～C ₁₄ Heterocycloalkyl, C ₁ ～C ₇ The heteroatoms in the heteroaryl group are independently selected from one or more of boron, silicon, oxygen, sulfur, selenium, nitrogen, and phosphorus; the number of heteroatoms is independently 1, 2, 3 or 4;

the pyrazolopyrimidine compound shown in the formula I is not

R ^1-1-4 independently halogen, -OH, -CN, amino, mercapto, C ₁ ～C ₇ Alkyl, C ₁ ～C ₇ Alkoxy, C ₁ ～C ₇ Alkylthio, C ₂ ～C ₇ Alkenyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl group,C ₆ ～C ₁₀ Aryl, C ₁ ～C ₇ Heteroaryl, or, substituted with 1 or 2R ^1-1-4-1 Substituted amino groups; r is R ^1-1-4-1 Independently C ₁ ～C ₇ Alkyl or C ₃ ～C ₁₄ Cycloalkyl;

R ^1-2 is hydrogen, -CN, -OR ^1-2-1 Or C ₁ ～C ₇ An alkyl group;

R ^1-4-3 independently a halogen,-OH, -CN, amino, mercapto, C ₁ ～C ₇ Alkyl, C ₁ ～C ₇ Alkoxy, C ₁ ～C ₇ Alkylthio, C ₂ ～C ₇ Alkenyl, C ₂ ～C ₇ Alkynyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl, C ₆ ～C ₁₀ Aryl, C ₁ ～C ₇ Heteroaryl, or, substituted with 1 or 2R ^1-4-3-1 Substituted amino groups; r is R ^1-4-3-1 Independently C ₁ ～C ₇ Alkyl or C ₃ ～C ₁₄ Cycloalkyl;

In one embodiment, certain substituents in the pyrazolopyrimidine compound of formula I or a pharmaceutically acceptable salt thereof may further have the following definitions, and the definitions of substituents not referred to below are as described in any of the above embodiments (hereinafter referred to simply as "in one embodiment"):

the said processIs->

In one embodiment:

when R is ¹ Optionally by 1, 2 or 3R ^1-5 Substituted C ₃ ～C ₁₄ In the case of heterocycloalkyl, said C ₃ ～C ₁₄ Heterocycloalkyl group For example C ₃ ～C ₁₄ Monocyclic heterocycloalkyl, C ₃ ～C ₁₄ Spirocyclic heterocycloalkyl, C ₃ ～C ₁₄ Condensed ring heterocycloalkyl or C ₃ ～C ₁₄ Bridged heterocycloalkyl.

Piperidyl radicals such asSaid oxetanyl group being for example +.>

In one embodiment:

when R is ^1-1 Is optionally substituted with 1R ^1-1-4 Substituted C ₁ ～C ₇ Alkyl, R ^1-1-4 In the case of-OH, the said "optionally substituted with 1R ^1-1-4 Substituted C ₁ ～C ₇ Alkyl "such as hydroxymethyl.

In one embodiment:

The C is ₃ ～C ₁₄ Monocyclic heterocycloalkyl, for example, "C having 1 or 2 hetero atoms selected from one or both of N, O and S ₃ ～C ₉ Monocyclic heterocycloalkyl ", for example also" C having 1 or 2 heteroatoms selected from one or both of N, O and S ₃ ～C ₅ Monocyclic heterocycloalkyl ", also for example" C having 1 or 2 hetero atoms selected from one or both of N, O and S ₃ ～C ₅ Monocyclic heterocycloalkyl ", and which is attached to the benzene ring through a carbon atom, such as, for example, oxetan, more for example oxetan-3-yl.

In one embodiment:

the atoms in the pyrazolopyrimidine compound shown in the formula I or the pharmaceutically acceptable salt thereof can exist in the form of natural abundance.

In one embodiment:

R ^1-2 hydrogen, -CN or-OH.

In one embodiment:

R ^1-5 independently halogen,)CN、-NR ^1-5-1 R ^1-5-2 、-(C＝O)R ^1-5-3 、C ₁ ～C ₇ Alkyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl or C ₁ ～C ₁₀ Heteroaryl;

In one embodiment:

R ^1-5-1 and R is ^1-5-2 Is hydrogen;

In one embodiment:

R ¹ is H, -C (=O) R ^1-1 、-C(＝NR ^1-2 )R ^1-3 、-S(＝O) ₂ R ^1-4 Or, optionally, by 1, 2 or 3R ^1-5 Substituted: c (C) ₁ ～C ₇ Alkyl, C ₃ ～C ₁₄ Cycloalkyl, C ₃ ～C ₁₄ Heterocycloalkyl or C ₁ ～C ₁₀ Heteroaryl group；

R ^1-2 hydrogen, -CN or-OH;

In one embodiment:

R ^1-5-1 and R is ^1-5-2 Is hydrogen;

In one embodiment:

R ^1-5-1 and R is ^1-5-2 Is hydrogen;

In a certain scheme, the pyrazolopyrimidine compound shown in the formula I or pharmaceutically acceptable salt thereof has any one of the following structures:

the invention also provides a preparation method of the compound I, which is any one of the following methods:

method 1: which comprises the following steps: step one, oxidizing a compound 1A in an organic solvent by using an oxidant to obtain a compound 1B; step two, reacting the compound 1B with the compound 1C under the conditions of an organic solvent and alkalinity to obtain a compound 1D; step three, removing the protecting group PG on the N of the compound 1D to obtain a compound 1E; step four, carrying out reductive amination reaction on the compound 1E and aldehyde or ketone in an organic solvent to obtain a compound I;

method 2: the compound 2A and the compound 1A undergo substitution reaction to obtain a compound I;

method 3: compound 1E and Compound R ^1-1 -COOH is subjected to condensation reaction to obtain a compound I;

Step two, the organic solvent is preferably dichloromethane or toluene; the basic conditions are preferably organic bases such as N, N-Diisopropylethylamine (DIPEA), triethylamine; the molar ratio of the compound 1B to the compound 1C, DEIPA is preferably 1:1:2, the reaction time is preferably 0 to 12 hours, and the reaction temperature is preferably 0 to 35 ℃.

Step three, the protecting group PG on N may be any suitable amino protecting group commonly used in the art, preferably Boc, with the aim of protecting compound 1C from certain reactive groups (e.g. amino groups) on it when it is involved in the reaction; the conditions for removing the protecting group PG on N may be conventional conditions for removing various protecting groups in the art, such as hydrolysis conditions, aminolysis conditions, hydrogenation conditions, etc.; if Boc is used as the protecting group, the deprotection reaction may be carried out under standard conditions, e.g., methylene chloride/trifluoroacetic acid system, saturated hydrogen chloride dioxane solution; after the reaction for removing the protecting group is finished, the method can further comprise post-treatment operation; the methods and conditions of the post-treatment may be conventional in the art for such post-reaction treatments, preferably: washing, drying, filtering and evaporating the solvent from the reaction system, and then performing column chromatography; or evaporating the reaction system to remove the solvent, washing and filtering; or evaporating the reaction system to remove the solvent, and performing thin layer chromatography.

The invention also provides a compound shown as a formula 1C or 2A:

wherein R is ¹ And PG are defined as above.

In one embodiment, the compound shown in formula 1C or 2A may be any one of the following compounds:

the invention also provides a preparation method of the compound shown in the formula 1C, which comprises the following steps: step one, in a solvent, under the action of alkali, carrying out substitution reaction on a compound 1C1 and halogenated nitrobenzene to obtain a compound 1C2; step two, in a solvent, reducing the compound 1C2 to obtain a compound 1C through reduction catalyst primordium;

In the first step, the solvent is preferably dimethylsulfoxide.

In the second step, the solvent is preferably methanol or ethanol.

The invention also provides a preparation method of the compound shown in the formula 2A, which is any one of the following methods:

in method 1: step one, carrying out reductive amination reaction on a compound 1C1 and aldehyde or ketone to obtain a compound 2A1; step two, removing the protecting group PG on N in the compound 2A1 to obtain a compound 2A2; step three, carrying out substitution reaction on the compound 2A2 and fluoronitrobenzene to obtain a compound 2A3; step four, reducing the compound 2A3 by a catalyst to obtain a compound 2A;

In method 2: step one, removing a protecting group PG on N in a compound 1C2 to obtain a compound 2A4; step two, carrying out substitution reaction on the compound 2A4 and a halogen substituted compound under alkaline conditions to obtain a compound 2A5, wherein the alkaline conditions are preferably potassium carbonate or silver oxide; the halogen substituted compound is preferably a bromide or an iodide; step three, reducing the compound 2A5 by a catalyst to obtain a compound 2A;

the substance X is pyrazolopyrimidine compound shown in the formula I or pharmaceutically acceptable salt thereof.

Unless otherwise indicated, the following terms appearing in the present specification and claims have the following meanings:

the term "pharmaceutically acceptable" refers to salts, solvents, excipients, and the like, which are generally non-toxic, safe, and suitable for patient use. The "patient" is preferably a mammal, more preferably a human.

The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention prepared with relatively non-toxic, pharmaceutically acceptable acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts may be obtained by contacting neutral forms of such compounds with a sufficient amount of a pharmaceutically acceptable base in pure solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include, but are not limited to: lithium salt, sodium salt, potassium salt, calcium salt, aluminum salt, magnesium salt, zinc salt, bismuth salt, ammonium salt, diethanolamine salt. When the compounds of the present invention contain relatively basic functional groups, the acid addition salts may be obtained by contacting the neutral form of such compounds with a sufficient amount of a pharmaceutically acceptable acid in pure solution or in a suitable inert solvent. The pharmaceutically acceptable acids include inorganic acids including, but not limited to: hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, phosphoric acid, phosphorous acid, sulfuric acid, and the like. The pharmaceutically acceptable acid includes organic acids including, but not limited to: acetic acid, propionic acid, oxalic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, salicylic acid, tartaric acid, methanesulfonic acid, isonicotinic acid, acidic citric acid, oleic acid, tannic acid, pantothenic acid, hydrogen tartrate, ascorbic acid, gentisic acid, fumaric acid, gluconic acid, sugar acid, formic acid, ethanesulfonic acid, pamoic acid (i.e., 4' -methylene-bis (3-hydroxy-2-naphthoic acid)), amino acids (e.g., glutamic acid, arginine), and the like. When the compounds of the present invention contain relatively acidic and relatively basic functional groups, they can be converted into base addition salts or acid addition salts. See, for example, berge et al, "Pharmaceutical Salts", journal of Pharmaceutical Science 66:1-19 (1977), or Handbook of Pharmaceutical Salts: properties, selection, and Use (P.Heinrich Stahl and Camille G.Wermuth, ed., wiley-VCH, 2002).

The term "solvate" refers to a substance formed by combining a compound of the invention with a stoichiometric or non-stoichiometric solvent. The solvent molecules in the solvate may be present in an ordered or unordered arrangement. Such solvents include, but are not limited to: water, methanol, ethanol, and the like.

The terms "pharmaceutically acceptable salts" and "solvates" in "solvates of pharmaceutically acceptable salts" refer, as described above, to compounds of the invention which are prepared from compound 1, with a relatively non-toxic, pharmaceutically acceptable acid or base, and 2, in combination with a stoichiometric or non-stoichiometric solvent. The "solvate of a pharmaceutically acceptable salt" includes, but is not limited to, the hydrochloride monohydrate of the compound of the invention.

The terms "compound," "pharmaceutically acceptable salt," "solvate," and "solvate of a pharmaceutically acceptable salt" may exist in crystalline or amorphous form. The term "crystalline form" refers to a form in which ions or molecules are strictly and periodically arranged in three dimensions in a defined manner and have a periodic recurrence pattern at certain intervals; due to the above-mentioned periodic arrangement, there may be various crystal forms, i.e., polymorphism. The term "amorphous" refers to a state in which ions or molecules are randomly distributed, i.e., the ions and molecules do not have a periodic arrangement rule.

The terms "compound," "pharmaceutically acceptable salt," "solvate," and "solvate of a pharmaceutically acceptable salt," if stereoisomers are present, may exist as single stereoisomers or as mixtures thereof (e.g., racemates). The term "stereoisomer" refers to a cis, trans or optical isomer. These stereoisomers may be isolated, purified and enriched by asymmetric synthesis methods or chiral separation methods (including but not limited to thin layer chromatography, rotary chromatography, column chromatography, gas chromatography, high pressure liquid chromatography, etc.), and may be obtained by chiral resolution by bonding (chemical bonding, etc.) or salifying (physical bonding, etc.) other chiral compounds. The term "single stereoisomer" means that one stereoisomer of the compound of the present invention is present in an amount of not less than 95% by mass relative to all stereoisomers of the compound.

The terms "compound," "pharmaceutically acceptable salt," "solvate," and "solvate of pharmaceutically acceptable salt," if present, may exist as a single tautomer or as a mixture thereof, preferably in the form of the more stable tautomer.

The atoms in the terms "compound," "pharmaceutically acceptable salt," "solvate," and "solvate of a pharmaceutically acceptable salt" may exist in either their natural or non-natural abundance. Taking a hydrogen atom as an example, the natural abundance form thereof refers to the case in which about 99.985% is protium and about 0.015% is deuterium; its non-naturally abundant form is for example deuterium in which about 95%. That is, one or more atoms in the terms "compound," "pharmaceutically acceptable salt," "solvate," and "solvate of a pharmaceutically acceptable salt" may be an atom that exists in a non-naturally abundant form.

When any variable (e.g. R ^1-1-1 ) In the definition of a compound, the definition of each position of the variable is independent of the definition of the other positions, and the meanings of the variable are independent and do not influence each other. Thus, if a group is substituted with 1, 2 or 3R ^1-1-1 The radical is substituted, that is to say, it may be substituted by up to 3R ^1-1-1 Substituted, at position R ^1-1-1 Definition of (d) and the remaining position R ^1-1-1 Are defined independently of each other. In addition, combinations of substituents and/or variables are allowed only if the combination yields a stable compound.

The term "optionally substituted" means that it may or may not be substituted.

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

The term "alkyl" refers to a saturated straight or branched monovalent hydrocarbon radical having one to twelve carbon atoms (e.g., C ₁ -C ₆ Alkyl radicals, e.g. C ₁ -C ₄ Alkyl). Examples of alkyl groups include, but are not limited to, methyl, ethyl, 1-propyl, 2-propyl, 1-butyl, 2-methyl-1-butyl, 2-methyl-2-propyl, 1-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl4-methyl-2-pentyl, 3-methyl-3-pentyl, 2, 3-dimethyl-2-butyl, 3-dimethyl-2-butyl, 1-heptyl and 1-octyl.

The term "alkenyl" refers to a compound having at least one unsaturated site, i.e., carbon-carbon sp ² Straight-chain or branched monovalent hydrocarbon radicals of two to twelve carbon atoms of the double bond (e.g. C ₂ -C ₆ Alkenyl radicals, also e.g. C ₂ -C ₄ Alkenyl) and includes groups having "cis" and "trans" orientations or "E" and "Z" orientations. Examples include, but are not limited to, vinyl, allyl.

The term "alkynyl" refers to a straight or branched monovalent hydrocarbon radical of two to twelve carbon atoms (e.g., C ₂ -C ₆ Alkynyl radicals, also e.g. C ₂ -C ₄ Alkynyl). Examples include, but are not limited to, ethynyl and propynyl.

The term "cycloalkyl" refers to a saturated or partially unsaturated (containing 1 or 2 double bonds) non-aromatic cyclic hydrocarbon radical having three to twenty carbon atoms (e.g., C ₃ -C ₆ Cycloalkyl), including monocyclic cycloalkyl and polycyclic cycloalkyl. Cycloalkyl groups contain 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 6 carbon atoms.

Examples of monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, 5-hexenyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl, and 1-cyclohex-3-enyl.

Polycyclic cycloalkyl groups are polycyclic (e.g., bicyclic and tricyclic) cycloalkyl structures including spiro, fused, and bridged cycloalkyl groups. Where cycloalkyl "of a spiro ring refers to a polycyclic group sharing one carbon atom (called a spiro atom) between 5-to 20-membered monocyclic rings, which may contain one or more double bonds, but no ring has a fully conjugated pi-electron system. Preferably 6 to 14 membered, more preferably 7 to 10 membered. Dividing spirocycloalkyl groups according to the number of common spiro atoms between rings Is a monocycloalkyl, a bisspirocycloalkyl or a multicycloalkyl group, preferably a monocycloalkyl or a bisspirocycloalkyl group. More preferably 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered monocyclocycloalkyl. Examples of spirocycloalkyl groups include, but are not limited to:"fused ring cycloalkyl" refers to a 5 to 20 membered, all carbon polycyclic group wherein each ring in the system shares an adjacent pair of carbon atoms with the other rings in the system, which may contain one or more double bonds, but no ring has a fully conjugated pi electron system. Preferably 6 to 14 membered, more preferably 7 to 10 membered. The number of constituent rings may be classified as a bicyclic, tricyclic, tetracyclic or polycyclic fused ring alkyl group, preferably a bicyclic or tricyclic, more preferably a 5-membered/5-membered or 5-membered/6-membered bicycloalkyl group. Examples of fused ring alkyl groups include, but are not limited to: "bridged ring cycloalkyl" refers to a 5 to 20 membered, all-carbon polycyclic group wherein any two rings share two carbon atoms that are not directly attached, which may contain one or more double bonds, but none of the rings has a fully conjugated pi-electron system. Preferably 6 to 14 membered, more preferably 7 to 10 membered. Cycloalkyl groups which may be classified as bicyclic, tricyclic, tetracyclic or polycyclic bridged according to the number of constituent rings are preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. Examples of bridged cycloalkyl groups include, but are not limited to: / >

The term "heterocycloalkyl" refers to a saturated carbocyclic group having 3 to 20 ring atoms, wherein at least one ring atom is a heteroatom independently selected from boron, silicon, oxygen, sulfur, selenium, nitrogen and phosphorusThe remaining ring atoms are C. The group may be a carbon group or a heteroatom group (i.e., it may be C-linked or N-linked, as long as it is possible). Examples of heterocyclyl groups include, but are not limited to, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, 4-thiomorpholinyl, thiaxalkyl, and piperazinyl. Fused ring moieties, spiro ring moieties, and bridged ring moieties are also included within the scope of this definition. For example, the groups derived from tetrahydropyrrole may be either tetrahydropyrrole-1-yl (N-linked) or tetrahydropyrrole-3-yl (C-linked). For example a monocyclic ring of 3-7 membered rings (1-6 carbon atoms and 1-3 heteroatoms selected from N, O, P, B, si, S and Se, where N, B, P or Se is optionally substituted by one or more oxygen atoms to give an image NO, BOH, PO, PO ₂ A group of SeO; n may optionally be quaternized; the S atom may be optionally substituted by one or more oxygen or nitrogen atoms to give a compound like SO, SO ₂ 、S(＝O)(＝NR ^a ),S(＝NR ^b ) Or S (=NR) ^c ) ₂ At the same time, R ^a 、R ^b And R is ^c Independently cyano, C ₁ ～C ₇ Alkyl, C ₃ ～C ₁₄ Cycloalkyl radicals, the hetero atoms of which are one or more of boron, silicon, oxygen, sulfur, selenium, nitrogen and phosphorus, and C with the hetero atoms of 1-4 ₃ ～C ₁₄ Heterocyclyl, C having 1-4 hetero atoms, which are one or more of boron, silicon, oxygen, sulfur, selenium, nitrogen and phosphorus ₁ ～C ₇ Heteroaryl ", C ₆ ～C ₁₀ Aryl or C ₁ ～C ₇ An alkoxy group; at the same time, -CH ₂ The-group may optionally be-C (=o) -, -C (=s) -or-C (=nr ^d ) -instead, R ^d Independently cyano, C ₁ ～C ₇ Alkyl, C ₃ ～C ₁₄ Cycloalkyl radicals, the hetero atoms of which are one or more of boron, silicon, oxygen, sulfur, selenium, nitrogen and phosphorus, and C with the hetero atoms of 1-4 ₃ ～C ₁₄ Heterocyclyl, C having 1-4 hetero atoms, which are one or more of boron, silicon, oxygen, sulfur, selenium, nitrogen and phosphorus ₁ ～C ₇ Heteroaryl ", C ₆ ～C ₁₀ Aryl or C ₁ ～C ₇ An alkoxy group; when the ring is a three-membered ring, only one of which is a heteroatom), or a bicyclic ring of 7-10 atoms (4-9 carbon atoms and 1-3 heteroatoms selected from N, O, P, B, si, S, where N, S, B or P is optionally substituted by one or more oxygen atoms to give a compound of formula NO, BOH, SO, SO ₂ ，PO，PO ₂ Groups of SeO, simultaneously, -CH ₂ The group may optionally be replaced by-C (=o) -. Depending on the structure, the heterocyclic group may be a monovalent group or a divalent group, i.e., a heterocyclylene group.

The term "aryl" refers to any stable mono-or bi-cyclic carbocycle of up to 10 atoms in each ring, wherein at least one ring is an aromatic ring. Examples of the above aryl unit include phenyl, naphthyl, tetrahydronaphthyl, 2, 3-indanyl, biphenyl, phenanthryl, anthracyl, or acenaphthyl (acenaphthyl). It will be appreciated that where the aryl substituent is a bicyclic substituent and one of the rings is a non-aromatic ring, the connection is through an aromatic ring.

The term "heteroaryl" refers to stable mono-or bi-rings of up to 7 atoms in each ring, wherein at least one ring is aromatic and contains 1-4 heteroatoms selected from boron, silicon, oxygen, sulfur, selenium, nitrogen and phosphorus. Heteroaryl groups within the scope of this definition include, but are not limited to: acridinyl, carbazolyl, cinnolinyl, quinoxalinyl, pyrazolyl, indolyl, benzotriazole, furanyl, thienyl, benzothienyl, benzofuranyl, quinolinyl, isoquinolinyl, oxazolyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridinyl, pyrimidinyl, pyrrolyl, tetrahydroquinolinyl. "heteroaryl" shall also be understood to include any N-oxide derivative of a nitrogen-containing heteroaryl. In the case where the heteroaryl substituent is a bicyclic substituent and one ring is a non-aromatic ring or contains no heteroatoms, it is understood that the linking occurs through the aromatic ring separately. The heteroaromatic ring systems may be cyclic in fused form. Wherein N, S, B, P or Se is optionally substituted with one or more oxygen atoms to give a compound like NO, SO ₂ 、BOH、PO、PO ₂ A group of SeO, an N atom being capable of being quaternaryAnd (5) ammonifying. Heteroaryl groups may be attached to the main structure at any heteroatom or carbon atom that results in the formation of a stable compound. Depending on the structure, heteroaryl groups may be monovalent or divalent, i.e., heteroarylene.

The term "alkoxy" refers to an alkyl group attached through an oxygen bridge; the definition of the alkyl is the same as that above.

The term "alkylthio" refers to an alkyl group attached through a sulfur bridge; the definition of the alkyl is the same as that above.

The term "component" refers to the individual components of the combination of the invention, namely compound I, a pharmaceutically acceptable salt thereof, a solvate of a pharmaceutically acceptable salt thereof, a metabolite thereof or a prodrug thereof, or an anticancer drug.

The term "pharmaceutical excipients" refers to excipients and additives used in the manufacture of medicaments and formulation of prescriptions, and is all matter contained in the pharmaceutical formulation except for the active ingredient. See the four pharmacopoeias of the people's republic of China (2015 Edition), or Handbook of Pharmaceutical Excipients (RaymondC Rowe,2009Sixth Edition).

The term "treatment" refers to therapeutic therapy. When specific conditions are involved, treatment refers to: (1) alleviating a disease or one or more biological manifestations of a disorder, (2) interfering with (a) one or more points in a biological cascade that results in or causes a disorder or (b) one or more biological manifestations of a disorder, (3) ameliorating one or more symptoms, effects, or side effects associated with a disorder, or one or more symptoms, effects, or side effects associated with a disorder or treatment thereof, or (4) slowing the progression of a disorder or one or more biological manifestations of a disorder.

The term "preventing" refers to a reduced risk of acquiring or developing a disease or disorder.

The term "therapeutically effective amount" refers to an amount of a compound that, when administered to a patient, is sufficient to effectively treat a disease or disorder described herein. The "therapeutically effective amount" will vary depending on the compound, the condition and severity thereof, and the age of the patient to be treated, but can be adjusted as desired by one of ordinary skill in the art.

The term "patient" refers to any animal, preferably a mammal, most preferably a human, that is about to or has received administration of the compound or composition according to embodiments of the present invention. The term "mammal" includes any mammal. Examples of mammals include, but are not limited to, cattle, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc., with humans being preferred.

The term "active ingredient" refers to the active ingredient in the pharmaceutical composition or combination kit of the invention, i.e. compound I, a pharmaceutically acceptable salt thereof, a solvate of a pharmaceutically acceptable salt thereof, a metabolite thereof or a prodrug thereof, an anticancer drug, or a combination of the above.

With wedge-shaped keys and broken-line keys Representing the absolute configuration of a stereogenic center.

The above preferred conditions can be arbitrarily combined on the basis of not deviating from the common knowledge in the art, and thus, each preferred embodiment of the present invention can be obtained.

The reagents and materials used in the present invention are commercially available.

The invention has the positive progress effects that: the compound of the invention has better inhibitory activity on WEE1 kinase and better bioavailability.

Detailed Description

The invention is further illustrated by means of the following examples, which are not intended to limit the scope of the invention. The experimental methods, in which specific conditions are not noted in the following examples, were selected according to conventional methods and conditions, or according to the commercial specifications.

The structures of all the compounds of the invention can be changed by nuclear magnetic resonance ¹ H NMR) and/or mass spectrometry detection (MS) identification. ¹ H NMR chemical shifts (delta) were recorded in PPM (10 ^-6 ). NMR was performed by Bruker AVANCE-400 spectrometer.

LC-MS was determined by Agilent 1200HPLC/6120 mass spectrometer.

The thin layer silica gel plate is a good minister silica source HSGF254 or Qingdao GF254 silica gel plate. Column chromatography generally uses 200-300 mesh silica gel of yellow sea as carrier.

Example 1 and example 39

The first step:

the compound (I-1-a) (3 aR,6 aS) -2, 3a,4,6 a-hexahydro-1H-pyrrolo [3,4-c ]Pyrrole-5-carboxylic acid tert-butyl ester (2000 mg,9.4211 mmol) was dissolved in DMSO (30 mL), then potassium carbonate (3 eq,28.263 mmol) and 1-fluoro-4-nitro-benzene (I-1-b) (1 eq,9.4211 mmol) were added and the reaction was heated to 120℃and stirred for 2 hours. The reaction was poured into 5ml of water, forming a yellow solid. The solid was filtered, washed with water (2X 5 ml), the solid was collected and dried in vacuo to give the compound (I-1-c) tert-butyl (3 aR,6 aS) -2- (4-nitrophenyl) -1, 3a,4,6 a-hexahydropyrrolo [3, 4-c)]Pyrrole-5-carboxylic acid tert-butyl ester (3 g,8.998 mmol), yield 95.51%, yellow solid. LC-MS: M/z (M+H-tBu) ⁺ ＝278.2。

And a second step of:

the compound (I-1-c) (3 aR,6 aS) -2- (4-nitrophenyl) -1, 3a,4,6 a-hexahydropyrrolo [3, 4-c)]Pyrrole-5-carboxylic acid tert-butyl ester (0.4 g,1 mmol) was dissolved in a solution of ethanol (70 mL) and THF (10 mL), then Pd/C (0.1 eq,0.1 mmol) and amine formate (10 eq,10 mmol) were added. The reaction was heated to reflux and stirred for 1 hour. The reaction was filtered and concentrated to give the compound (I-1-d) (3 aR,6 aS) -2- (4-aminophenyl) -1, 3a,4,6 a-hexahydropyrrolo [3, 4-c)]Pyrrole-5-carboxylic acid tert-butyl ester (0.36 g,1.2 mmol), yield 100% as a black solid. LC-MS: M/z (M+H) ⁺ ＝304.2。

And a third step of:

the compound (I-1-e) 2-allyl-1- [6- (1-hydroxy-1-methyl-ethyl) -2-pyridinyl ]-6-methylsulfanyl-pyrazolo [3,4-d ]]Pyrimidine-3-one (317 mg,0.9989 mmol) was dissolved in toluene (15 mL), and then mCPBA (1.2 eq,1.199mmol, 77%) was added and the reaction stirred at room temperature for 2 hours. Then, the compound (I-1-d) (3 aR,6 aS) -2- (4-aminophenyl) -1 is added into the reaction liquid in sequence,3,3a,4,6 a-hexahydropyrrolo [3,4-c ]]A solution of pyrrole-5-carboxylic acid tert-butyl ester (1.2 eq,0.18 mmol) in toluene (2 mL) and DIPEA (2 eq,1.998 mmol) was reacted further at room temperature with stirring for 12 hours. The reaction was quenched with water and extracted with ethyl acetate (2X 20 ml), the organic phase was washed with brine (1X 20 ml) and dried over Na ₂ SO ₄ Drying, filtering and concentrating to obtain crude product. The crude product was washed once with methanol 2ML, filtered and the compound (I-39) (3 ar,6 as) -2- [4- [ [ 2-allyl-1- [6- (1-hydroxy-1-methyl-ethyl) -2- ] was obtained]Tert-butyl pyridyl]-3-oxo-pyrazolo [3,4-d]Pyrimidin-6-yl]Amino group]Phenyl group]-1, 3a,4,6 a-hexahydropyrrolo [3,4-c]Pyrrole-5-carboxylate (320 mg,0.5223 mmol), yield 52.29%, yellow solid. ¹ H NMR(400MHz,MeOD)δ8.77(s,1H),7.97(t,J＝7.9Hz,1H),7.80(s,1H),7.64(d,J＝7.7Hz,1H),7.47(d,J＝7.8Hz,1H),6.60(d,J＝9.0Hz,2H),5.80–5.66(m,1H),5.05(dd,J＝10.2,1.1Hz,1H),4.93(dd,J＝17.1,1.3Hz,1H),4.82(d,J＝6.1Hz,2H),3.67(s,2H),3.56–3.47(m,2H),3.38(d,J＝8.6Hz,1H),3.30(s,1H),3.23(dd,J＝9.8,3.5Hz,2H),3.07(s,2H),1.59(s,6H),1.52–1.43(m,9H).LC-MS:m/z:(M+H) ⁺ ＝613.3。

Fourth step:

the compound (I-39) (3 aR,6 aS) -2- [4- [ [ 2-allyl-1- [6- (1-hydroxy-1-methyl-ethyl) -2-pyridinyl]-3-oxo-tert-butylpyrazolo [3,4-d ] ]Pyrimidin-6-yl]Amino group]Phenyl group]-1, 3a,4,6 a-hexahydropyrrolo [3,4-c]Pyrrole-5-carboxylate (320 mg,0.52 mmol) was dissolved in dichloromethane (5 mL) and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 12 hours. The reaction was concentrated and saturated NaHCO was added ₃ The solution was dissolved and then extracted with ethyl acetate (2×40 ml). The organic layer was taken up with Na ₂ SO ₄ Drying, filtering and concentrating to obtain compound (I-1) 6- [4- [ (3 aR,6 As) -2, 3a,4,6 a-hexahydro-1H-pyrrolo [3, 4-c)]Pyrrol-5-yl]Anilino group]-2-allyl-1- [6- (1-hydroxy-1-methyl-ethyl) -2-pyridinyl]Pyrazolo [3,4-d]Pyrimidin-3-one (110 mg,0.2146 mmol) in 41.09% yield as yellow solid. LC-MS: M/z (M+H) ⁺ ＝513.3。

Example 2

The compound (I-1) 6- [4- [ (3 aR,6 aS) -2, 3a,4,6 a-hexahydro-1H-pyrrolo [3,4-c]Pyrrol-5-yl]Anilino group]-2-allyl-1- [6- (1-hydroxy-1-methyl-ethyl) -2-pyridinyl]Pyrazolo [3,4-d]Pyrimidine-3-one (110 mg,0.2146 mmol) was dissolved in methanol solution (6 mL), then formaldehyde (10 eq,2.146mmol, 37%) and glacial acetic acid (0.2 mL) were added and the reaction stirred at room temperature for 5 min. Then, sodium borohydride acetate (2 eq,0.4292 mmol) was added thereto, and the reaction solution was stirred at room temperature for further reaction for 12 hours. NaHCO for reaction ₃ The aqueous solution was quenched and extracted with ethyl acetate (2X 20 ML), the organic layer was washed with brine (1X 20 ML) and with Na ₂ SO ₄ Drying, filtering and concentrating to obtain a crude product. The crude product was further purified by Pre-HPLC using CH ₃ CN/water (0.1% HCOOH) was eluted from 20% to 50% to give the compound (I-2) 6- [4- [ (3 aR,6 aS) -2-methyl-1, 3a,4,6 a-hexahydro-hexahydropyrrolo [3,4-c ]]Pyrrol-5-yl]Anilino group]-2-allyl-1- [6- (1-hydroxy-1-methyl-ethyl) -2-pyridinyl]Pyrazolo [3,4-d]Pyrimidin-3-one (40 mg,0.07596 mmol) in 35.40% yield as yellow solid. ¹ H NMR(400MHz,MeOD)δ8.80(s,1H),8.42(s,1H),7.97(t,J＝7.9Hz,1H),7.78(d,J＝8.0Hz,1H),7.65(d,J＝7.7Hz,1H),7.54(d,J＝8.7Hz,2H),6.82(d,J＝9.0Hz,2H),5.79–5.67(m,1H),5.05(dd,J＝10.2,1.2Hz,1H),4.93(dd,J＝17.1,1.3Hz,1H),4.81(s,2H),3.64(s,2H),3.55(d,J＝10.0Hz,2H),3.29(s,4H),3.15(dd,J＝9.9,5.7Hz,2H),2.92(s,3H),1.58(d,J＝8.3Hz,6H).LC-MS:m/z:(M+H) ⁺ ＝527.3。

Example 15

The first step:

1g of tert-butyl (3 aR,6 aS) -5- (4-nitrophenyl) hexahydropyrrole [3,4-c]Pyrrole-2 (1H) -carboxylate (I-1-c) was dissolved in 40ml of methylene chloride, 10ml of trifluoroacetic acid was added thereto, and the reaction mixture was swirled after 1 hour of reaction at room temperatureDried to give 0.7g of yellow solid (I-15-a) in 91% yield. LC-MS: M/z (M+H) ⁺ ＝234.1。

And a second step of:

(3 aR,6 aS) -2- (4-nitrophenyl) octahydro pyrrole [3,4-c]Pyrrole (I-15-a) (200 mg,0.26 mmol) was dissolved in 15ml of acetonitrile, and potassium carbonate (170 mg,1.72 mmol) and bromoacetonitrile (I-15-b) (124 mg,1 mmol) were added. After 18h reaction at room temperature, the mixture was concentrated and extracted with 30ml of dichloromethane and 10ml of water. By thin layer chromatography (DCM/CH ₃ OH=10/1) to give 40mg of yellow solid (I-15-c) in 56% yield. LC-MS: M/z (M+H) ⁺ ＝273.1。

And a third step of:

2- ((3 AR,6 AS) -5- (4-nitrophenyl) -3a,6 a-dimethylhexahydropyrrolo [3, 4-c)]Pyrrole-2 (1H) -yl) acetonitrile (I-15-c) (50 mg,0.18 mmol) was dissolved in 5ml of tetrahydrofuran and reacted at room temperature under an atmosphere of hydrogen for half an hour. The reaction solution was filtered and concentrated to give 42mg of pale-tea-colored solid (I-15-d) in 94% yield. LC-MS: M/z (M+H) ⁺ ＝243.2。

Fourth step:

2-allyl-1- (6- (2-hydroxypropan-2-yl) pyridin-2-yl) -6- (methylsulfanyl) -1, 2-dihydro-3H-pyrazolo [3,4-d]Pyrimidin-3-one (I-1-e) (850 mg,0.14 mmol) was dissolved in 10ml of toluene, 3-chloroperoxybenzoic acid (38 mg,0.17 mol) was added thereto, and the mixture was stirred at room temperature for 20 minutes and then dried by spin. Adding 2- ((3 AR,6 AS) -5- (4-aniline) -3a,6 a-dimethyl hexahydropyrrolo [3, 4-c)]Pyrrole-2 (1H) -yl) acetonitrile (I-15-d) (742 mg) was dissolved in 5ml dimethyl sulfoxide, triethylamine (28 mg,0.28 mmol) was added, heated to 60℃and stirred for about 18 hours. The reaction solution was cooled, extracted with water and ethyl acetate, concentrated and purified by thin layer chromatography (DCM/CH ₃ OH/NH ₃ .CH ₃ OH=100/10/1.5) to yield 24mg of yellow solid (I-15) in 32% yield. ¹ H NMR(400MHz,Methanol-d ₄ )δ8.78(s,1H),8.06–7.88(m,1H),7.78(d,J＝8.1Hz,1H),7.64(dd,J＝7.7,0.9Hz,1H),7.48(d,J＝8.4Hz,2H),6.82–6.63(m,2H),5.73(ddt,J＝17.1,10.2,6.1Hz,1H),5.05(dq,J＝10.2,1.2Hz,1H),4.97–4.89(m,2H),4.82(dt,J＝6.2,1.3Hz,2H),3.40(dd,J＝9.5,7.5Hz,2H),3.19(dd,J＝9.5,3.0Hz,2H),3.09–2.98(m,2H),2.94(dd,J＝9.0,7.1Hz,2H),2.65(dd,J＝9.3,3.4Hz,2H),1.59(s,6H).LC-MS:m/z:(M+H) ⁺ ＝552.3。

Example 32

2-allyl-6- (((4- ((3 ar,6 as) -5- (azetidin-3-yl) hexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) phenyl) amino) -1- (6- (2-hydroxypropan-2-yl) -1, 2-dihydro-3H-pyrazolo [3,4-d ] pyrimidin-3-one (I-33) (41 mg,0.072 mmol) was dissolved in methanol (10 mL), then formaldehyde solution (0.1 mL) and sodium triacetyl borohydride (46 mg,0.216 mmol) were added, the reaction mixture was stirred at room temperature for 16 hours, a mixture of dichloromethane and methanol (10/1, 20 mL) was dissolved, saturated sodium bicarbonate was washed twice, the organic layer was dried over anhydrous sodium sulfate, and the crude product obtained was concentrated by thin layer chromatography, and purified (7M ammonia methanol: dichloromethane=0-10%) to give the target compound (I-32) 24mg, 57% [ 1 mhz ], 8.9hz (7.96 hz), 7.9hz (7.19 hz), 7.7 hz (6 hz), 7.96 hz (7.7H), 7.216 hz), 7.7H (7 mg, 7 h=7.96 hz), 7.19 hz (7H), 7.7H), 7 h=7.96 hz (7.7H), 7H (7.96 hz (7H), 7.7H), 7H 1H), 7.7H 1, 7H (7.7H) and 1H) was obtained. 4.76 (d, j=6.1 hz, 2H), 3.76 (s, 2H), 3.48-3.32 (M, 2H), 3.22 (dd, j=15.5, 6.3hz, 5H), 3.00 (s, 2H), 2.88-2.75 (M, 2H), 2.52 (s, 3H), 2.36 (d, j=5.2 hz, 2H), 1.61 (s, 7H), LC-MS: M/z (m+h) +=582.3.

Example 33

2-allyl-6- ((4- (((3 aR,6 aS) -hexahydropyrrolo [3, 4-c))]Pyrrol-2 (1H) -yl) phenyl) -amino) -1- (6- (2-hydroxypropan-2-yl) pyridinyl-2-yl) -1, 2-dihydro-3H-pyrazolo [3,4-d]Pyrimidine-3-one (I-1) (80 mg,0.156 mmol) was dissolved in methanol (10 ml), then tert-butyl 3-oxaazetidine-1-carboxylate (267 mg,1.56 mmol) and sodium triacetylborohydride (66 mg,0.312 mmol) were added and stirred at room temperature for 18 hours. The reaction solution was concentrated, and concentrated with dichloromethane and methyl acetateThe alcohol mixture (10/1, 20 mL) was dissolved, the saturated sodium bicarbonate was washed twice, the organic layer was dried over anhydrous sodium sulfate, filtered, and the crude product obtained by concentration was isolated and purified by thin layer chromatography (7M methanolic ammonia: dichloromethane=0-10%) to give the Boc-protected target compound, which was dissolved in 20mL of dichloromethane, added with 0.5mL of 4N hydrogen chloride dioxane solution and stirred at room temperature for 16h. Concentrating under reduced pressure, dissolving the crude product with water, and extracting with dichloromethane. The aqueous layer was adjusted to ph=10 with 2N NaOH solution and filtered to give 47mg of the target compound (I-33) as a yellow solid in 53% yield. ¹ H NMR(400MHz,MeOD)δ8.85–8.72(m,1H),7.97(t,J＝7.9Hz,1H),7.78(d,J＝7.8Hz,1H),7.64(dd,J＝7.7,0.7Hz,1H),7.48(d,J＝8.3Hz,2H),6.70(d,J＝9.0Hz,2H),5.72(ddt,J＝16.4,10.3,6.1Hz,1H),5.05(dd,J＝10.2,1.1Hz,1H),4.94(d,J＝1.3Hz,1H),4.82(d,J＝6.1Hz,1H),3.80–3.44(m,2H),3.44–3.36(m,0H),3.31–3.15(m,2H),2.97(s,1H),2.89–2.71(m,1H),2.37(dd,J＝9.1,4.1Hz,1H),1.59(s,3H).LC-MS:m/z:(M+H) ⁺ ＝568.3。

Example 35

The first step:

6- [4- [ (3 aR,6 aS) -2, 3a,4,6 a-hexahydro-1H-pyrrolo [3,4-c]Pyrrol-5-yl]Anilino group ]-2-allyl-1- [6- (1-hydroxy-1-methyl-ethyl) -2-pyridinyl]Pyrazolo [3,4-d]Pyrimidine-3-one (I-1) (80 mg,0.1561 mmol) was dissolved in methanol solution (6 mL), and tert-butyl N- (3-oxocyclobutyl) carbamate (I-35-a) (10 eq,1.561 mmol) and sodium borohydride acetate (2 eq,0.3121 mmol) were added and the reaction stirred at room temperature for 12 hours. The reaction was quenched with saturated aqueous sodium bicarbonate, extracted with ethyl acetate (2X 20 ML), the organic layer was washed with brine (1X 20 ML), and dried over Na ₂ SO ₄ Drying, filtering and concentrating to obtain N- [3- [ (3 aS,6 aR) -tert-butyl-2- [4- [ [ 2-allyl-1- [6- (1-hydroxy-1-methyl-ethyl) -2-pyridyl)]-3-oxo-pyrazolo [3,4-d]Pyrimidin-6-yl]Amino group]Phenyl group]-1, 3a,4,6 a-hexahydropyrrolo [3,4-c]Pyrrol-5-yl]Cyclobutyl group]50mg of carbamate (I-35-b), yellowThe yield of the coloured solid was 46.99%. LC-MS: M/z (M+H) ⁺ ＝682.5。

And a second step of:

n- [3- [ (3 aS,6 aR) -2- [4- [ [ 2-allyl-1- [6- (1-hydroxy-1-methyl-ethyl) -2-pyridinyl]T-butyl solution of (2)]-3-oxo-pyrazolo [3,4-d]Pyrimidin-6-yl]Amino group]Phenyl group]-1, 3a,4,6 a-hexahydropyrrolo [3,4-c]The pyrrol-5-yl group was dissolved in a solution of cyclohexylcarbamate (I-35-b) (50 mg,0.07334 mmol) in dichloromethane (2 mL) and TFA (1 mL) was added. The reaction was stirred at room temperature for 12 hours. The product was quenched with saturated aqueous sodium bicarbonate and concentrated to give the crude product. The crude product was purified by preparative HPLC eluting with acetonitrile/water (0.1% formic acid) from 20% to 50% to give 6- [4- [ (3 as,6 ar) -5- (3-aminocyclobutyl) -1,3 ]3A,4, 6a hexahydropyrrolo [3,4-c]Pyrrol-2-yl]Anilino group]-2-allyl-1- [6- (1-hydroxy-1-methyl-ethyl) -2-pyridinyl]Pyrazolo [3,4-d]Pyrimidine-3-one (I-35) 15mg, yellow solid, yield 35.16%. ¹ HNMR(400MHz,MeOD)δ8.79(s,1H),7.97(t,J＝7.9Hz,1H),7.79(d,J＝7.0Hz,1H),7.65(d,J＝7.7Hz,1H),7.50(d,J＝9.2Hz,2H),6.74(d,J＝9.0Hz,2H),5.77–5.67(m,1H),5.05(d,J＝9.0Hz,1H),4.95(d,J＝1.3Hz,1H),4.83(d,J＝6.1Hz,2H),3.30(s,0H),3.03(s,2H),2.90(dd,J＝25.5,8.3Hz,3H),2.56(s,2H),2.45(dd,J＝10.5,6.8Hz,3H),2.28(s,2H),2.13–2.00(m,2H),1.59(s,6H).LC-MS:m/z:(M+H) ⁺ ＝582.3。

Example 41

The compound (I-1) 6- [4- [ (3 AR,6 AS) -2, 3a,4,6 a-hexahydro-1H-pyrrolo [3,4-c]Pyrrol-5-yl]Anilino group]-2-allyl-1- [6- (1-hydroxy-1-methyl-ethyl) -2-pyridinyl]Pyrazolo [3,4-d]Pyrimidine-3-one (100 mg,0.1596 mmol) was dissolved in dichloromethane (5 mL), DIPEA (2 equiv.,0.3192 mmol) was added and the reaction stirred at room temperature for 5 min. 2-Hydroxyacetic acid (1.3 equiv.; 0.2075 mmol) and HATU (1 equiv.; 0.1596 mmol) were added in this order to the reaction mixture, and stirring was continued at room temperature for 12 hours. NaHCO for reaction ₃ Water-solubleThe solution was quenched and extracted with dichloromethane (2X 20 ML), the organic layer was washed with brine (1X 20 ML) and with Na ₂ SO ₄ Drying, filtering and concentrating to obtain a crude product. The crude product was further purified by normal phase silica gel column eluting with methanol/dichloromethane from 0% to 8% to give the compound (I-41) 6- [4- [ (3 AR,6 AS) -5- (2-hydroxyacetyl) -1, 3a,4,6 a-hexahydropyrrolo [3,4-c ]]Pyrrol-2-yl]Anilino group]-2-allyl-1- [6- (1-hydroxy-1-methyl-ethyl) -2-pyridinyl ]Pyrazolo [3,4-d]Pyrimidin-3-one (50 mg,0.08763 mmol) in 54.9% yield as yellow solid. ¹ H NMR(400MHz,MeOD)δ8.78(s,0H),7.97(t,J＝7.9Hz,1H),7.80(s,1H),7.64(dd,J＝7.7,0.8Hz,1H),7.47(s,2H),6.61(d,J＝9.0Hz,2H),5.78–5.67(m,1H),5.09–5.01(m,1H),4.94(d,J＝1.3Hz,1H),4.82(d,J＝6.1Hz,2H),4.19(s,2H),3.85–3.74(m,2H),3.53(ddd,J＝14.2,10.7,4.0Hz,3H),3.42(dd,J＝10.8,4.9Hz,1H),3.30–3.23(m,2H),3.16(dd,J＝25.6,13.0Hz,3H),1.59(s,6H).LC-MS:m/z:(M+H) ⁺ ＝571.3。

Example 47

The first step:

tert-pyrrolidinyl hexahydro [3,4-c ]]Pyrrole-2 (1H) -carboxylate (2.36 mmol) (I-47-a) was dissolved in methylene chloride (20 mL), and formaldehyde (11.8 mmol) and sodium triacetoxyborohydride (7.07 mmol) were added to the reaction solution, which was stirred at room temperature for 16 hours. Adding aqueous potassium carbonate solution to the reaction solution to adjust pH=9, extracting with dichloromethane, washing an organic phase with saturated saline solution, drying with anhydrous sodium sulfate, filtering, and evaporating filtrate to obtain a crude product of the target compound 5-methyl hexahydropyrrolo [3,4-c ]]Pyrrole-2 (1H) -carboxylic acid tert-butyl ester (I-47-b) (480 mg, 90.0%) was a colorless oil. LC-MS: M/z (M+H) ⁺ ＝227.2。

And a second step of:

5-Methylhexahydropyrrolo [3,4-c]Pyrrole-2 (1H) -carboxylic acid tert-butyl ester (I-47-b) (2.12 mmol) was dissolved in dichloromethane (10 mL), trifluoroacetic acid (10 mL) was added, and the reaction solution was stirred at room temperature for 0.5H. Evaporating the reaction solution to obtain crude productTarget compound 2-methyl octahydropyrrolo [3,4-c]Pyrrole 2, 2-trifluoroacetate salt (I-47-c) (250 mg, 93.4%) as a yellow solid. LC-MS: M/z (M+H) ⁺ ＝127.2。

And a third step of:

2-Methyloctahydropyrrolo [3,4-c ]]Pyrrole 2, 2-trifluoroacetate (1.98 mmol) (I-47-c) was dissolved in acetonitrile (20 mL), and potassium carbonate (5.94 mmol) and 1-fluoro-4-nitrobenzene (I-1-b) (2.97 mmol) were added to the reaction solution, and the reaction solution was heated to reflux and stirred for 16 hours. Filtering the reaction solution, evaporating filtrate to obtain a crude product, and purifying the crude product by column chromatography (dichloromethane/methanol=100/0-95/5) to obtain a target compound 2-methyl-5- (4-nitrophenyl) octahydropyrrolo [3, 4-c) ]Pyrrole (I-47-d) (250 mgg, 51.0%) as a yellow solid. LC-MS: M/z (M+H) ⁺ ＝248.3。

Fourth step:

120mg (0.48 mmol) of 2-methyl-5- (4-nitrophenyl) -1, 3a,4,6 a-hexahydropyrrolo [3,4-c]Pyrrole was dissolved in 5ml of methanol, then 10mg (10%) of Pd/C was added thereto and stirred at room temperature under a hydrogen atmosphere for 0.5 hours. The reaction liquid is filtered and concentrated to obtain 4- (5-methyl hexahydropyrrolo [3, 4-c)]Pyrrole-2 (1H) -yl) aniline (I-47-e) 100mg, 94% yield, pale tea solid. LC-MS: M/z (M+H) ⁺ ＝218.3。

Fifth step:

206mg (0.55 mmol) of 2-allyl-1- [6- (3-hydroxyoxetan-3-yl) -2-pyridinyl are reacted]-6-methylthiopyrazolo [3,4-d ]]Pyrimidine-3-ketolysis (I-47-f) was carried out in 10ml of toluene, 103mg (0.60 mol) of 3-chloroperoxybenzoic acid was added, stirring at room temperature for about 1 hour, and then 100mg (0.46 mmol) of 4- (5-methylhexahydropyrrolo [3, 4-c) were added]Pyrrole-2 (1H) -yl) aniline (formula I-47-e) and 120mg (0.92 mmol) DIPEA, heating to 30deg.C and stirring for about 16 hours, concentrating the reaction solution, and performing column chromatography (DCM/CH) ₃ Oh=10/1) followed by thin layer chromatography (DCM/CH ₃ OH/NH ₃ .CH ₃ After three purification steps of oh=100/10/1.5, 2-allyl-1- (6- (3-hydroxyoxetan-3-yl) pyridin-2-yl) -6- ((4- (5-methylhexahydropyrrolo [3, 4-c) is obtained ]Pyrrol-2 (1H) -yl) phenyl) amino) -1, 2-dihydro-3H-pyrazolo [3,4-d]Pyrimidine-3-one (I-47) 800mg, 32% yield, yellow solid. ¹ H NMR(400MHz,CDCl3)δ8.82(s,0H),8.03(t,J＝7.9Hz,1H),7.88(d,J＝8.0Hz,1H),7.39(d,J＝8.2Hz,2H),6.67(dd,J＝9.0,2.1Hz,1H),5.73(ddt,J＝16.3,10.2,6.1Hz,1H),5.14–5.06(m,2H),4.98(dd,J＝17.1,1.2Hz,1H),4.80(d,J＝7.2Hz,1H),4.67(d,J＝6.0Hz,1H),3.41(s,1H),3.25(dd,J＝9.4,2.7Hz,1H),3.03(d,J＝2.5Hz,1H),2.86–2.76(m,1H),2.51(s,1H),2.38(s,2H).LC-MS:m/z:(M+H) ⁺ ＝541.3。

Example 57

The first step:

to a solution of tert-butyl (3 ar,6 as) -5-oxahexahydrocyclopenta [ c ] pyrrole-2 (1 h) -carboxylate (I-57-a) in methanol (5 ml) was added a solution of dimethylamine in tetrahydrofuran (2M, 1.07mmol,0.54 ml), followed by addition of sodium borohydride acetate (566 mg,1.07 mmol) and stirring at room temperature for 16 hours. The reaction solution was dried in vacuo, saturated potassium carbonate solution (10 ml) and dichloromethane (3×10 ml) were added and extracted. The organic layer was dried over sodium sulfate solid and filtered. The organic layer solution was concentrated to give yellow solid I-57-b (158 mg, 70%).

And a second step of:

to a methanol (5 ml) solution of (3 ar,6 as) -n, n-dimethyldihydropyran [ c ] pyrrol-5-amine (I-57-b) was added trifluoroacetic acid (5 ml), and the mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated to give a pale brown crude product I-57-c (94 mg, 99%).

And a third step of:

to a solution of (3 ar,6 as) -n, n-dimethyldihydropyran [ c ] pyrrol-5-amine (I-57-c) (94 mg,0.625 mmol) in acetonitrile (5 ml) was added potassium carbonate (862mg, 0.625 mmol), and after stirring at room temperature for 10 minutes, p-fluoronitrobenzene (88 mg,0.625 mmol) was added. The reaction solution was stirred under reflux for 16 hours. After the reaction was cooled, it was concentrated, and extracted with water (20 ml) and dichloromethane (3×10 ml). The organic layer was dried over sodium sulfate solid and filtered. The organic layer solution was concentrated to give yellow solid I-57-d (66 mg, 42%).

Fourth step:

to a solution of (3 ar,6 as) -n, n-dimethyl-2- (4-nitrophenyl) octahydrocyclopenta [ c ] pyrrol-5-amine (I-57-d) (66 mg,0.27 mmol) in methanol (5 ml) was added palladium on carbon (20 mg, 10%) and the reaction mixture was stirred at room temperature under hydrogen for 16 hours. The reaction solution was filtered and concentrated to give light brown solid I-57-e (30 mg, 56%).

Fifth step:

to a solution of 2-allyl-1- (6- (2-hydroxypropyl-2-yl) pyridin-2-yl) -6- (methylsulfanyl) -1, 2-dihydro-3 h-pyrazolo [3,4-d ] pyrimidin-3-one (I-1-h) in toluene (3 ml) was added m-chloroperoxybenzoic acid (34 mg,0.166 mmol) and the mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated to a solid, which was then dissolved in dimethyl sulfoxide, followed by addition of Compound I-57-e (30 mg,0.138 mmol) and trifluoroacetic acid (0.02 ml), followed by stirring at 60℃for 16 hours. After the reaction was cooled to room temperature, it was extracted with water (5 ml) and dichloromethane (3×10 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated. The crude product was isolated by thin layer chromatography (dichloromethane: ethyl acetate: methanolic ammonia (7M) =10:4:1) to give I-57 (8 mg, 10%) as a pale yellow solid.

Referring to the above examples, the compounds shown in table 1 were prepared, and their structural characterization is as follows:

table 1 list of compounds

Effect example 1

1. In vitro inhibitory action of Compounds on WEE1 kinase

The testing method comprises the following steps:

test compounds were screened on WEE1 kinase using ELISA at ATP concentration of Km. Screening of 3 compounds on WEE1 kinase was performed to evaluate the kinase inhibitory activity of the test compounds. In the detection process, the initial concentration of the tested compound is 100nM, 6 gradient dilution concentrations are selected for each compound, the gradient dilution multiple is 4 times, and detection is carried out by 2 compound wells per concentration, and MK1775 is used as a standard control.

WEE1, available from Carna biosciences, inc., cat: 05-177; dimethyl sulfoxide, available from Sigma-Aldrich under the designation: d8418; ATP, purchased from Sigma-Aldrich, cat#: a7699; DTT solution, purchased from Sigma-Aldrich, cat number: 43816; protein Tyrosine Kinase (PTK) substrate (poly-Glu-Tyr), available from Sigma-Aldrich under the designation: p4476; P-Tyr (PY 99), available from Santa Cruz, cat: sc-7020; anti-mouse IgG HRP-linked Anti-body, available from Santa Cruz, cat: 7076S; TMB liquid Substrate System from Sigma-Aldrich, cat: t0440; costar Stripwell Microplate No Lid 1X 8Flat Bottom,Certified High Binding, from Sigma-Aldrich, cat: 42592; 96-well compound plate, purchased from Thermo Scientific, cat: 267245.

The testing steps are as follows:

1. coating a substrate: 1) An appropriate volume of substrate stock Protein Tyrosine Kinase (PTK) substrate (poly-Glu-Tyr) was taken, diluted 10-fold with PBS, and the concentration was diluted from 250mg/mL to 25mg/mL. Add to high adsorption 96 well plates, 125 μl per well. The mixture was placed in an incubator at 37℃overnight for coating. 2) After 24 hours, the 96-well plate is taken out, the liquid in the 96-well plate is poured out, the 96-well plate is washed 3 times by using a washing buffer, and the incubator at 37 ℃ is inverted and dried for 2 hours.

2. Preparation and transfer of the compound: 1) Compound dilution: taking 10mM of test compound stock solution, diluting the compound in multiple steps by using DMSO in a 96-well compound plate to obtain a compound with an initial concentration of 100X, and then taking the compound with the concentration as the first concentration, and carrying out 4-time gradient dilution by using DMSO for 6 concentrations; then respectively taking 2 mu L of gradient diluent and adding the gradient diluent into 48 mu L of 1 Xreaction buffer solution to prepare 4 Xcompounds for later use; 2) Transfer of 4 x compound: transferring 10 μl of 4×compound from the 96-well compound plate prepared in the above step into a dried high adsorption 96-well plate; 10 μl of the following liquids were added to the compound-free control wells and ATP-control wells: mu.L of DMSO was added to 48. Mu.L of 1 Xreaction buffer.

3. Enzyme reaction stage: 1) WEE1 kinase and ATP were prepared as a 2X enzyme solution and a 4X ATP solution in a 1X reaction buffer, respectively. Wherein in this screen, the final concentration of WEE1 kinase is: 0.15 ng/. Mu.L, final ATP concentration: 12. Mu.M; 2) 20. Mu.L of 2 enzyme solution was added to the high adsorption 96 well plate; 3) To the high adsorption 96-well plate, 10. Mu.L of 4 XATP solution was added, and 10. Mu.L of 1 Xreaction buffer was added to the ATP-control blank; 4) After centrifugation at 2000rpm for 20s in a HERAEUS Multifuge X R centrifuge, the plates were left to react at room temperature for 60min.

4. Reaction termination stage: 1) Pouring out the reaction liquid in the plate, adding 200 mu L of washing buffer into each hole, and washing for 5 times; primary antibody P-Tyr (PY 99) (dilution ratio 1:2000) was added at 100 μl per well for 30min at room temperature. 2) The primary antibody in the plate was removed, 200. Mu.L of washing buffer was added to each well, and the plate was washed 5 times; secondary Antibody-mouse IgG HRP-linked Antibody (dilution ratio 1:2000) was added at 100. Mu.L per well for 30min at room temperature. 3) The secondary antibody in the plate is poured out, washed 5 times by a washing buffer, TMB is added, and the color development is carried out for 10-30 min according to the color depth at each hole of 100 mu L. The reaction was quenched with 1N sulfuric acid prior to reading.

5. Detection and data processing: 1) Light absorption at wavelength 450nM is read on ThermoScientific MultiScan GO while background is read at 650 nM. 2) Log (inhibitor) vs. response-Variable slope (four) was performed on the data using Graphpad Prism 5.0 parameters) curve fitting, calculating the corresponding IC ₅₀ (half maximal inhibitory concentration)。

2. Test result data

The structure of the control sample used in the test is shown in Table 2.

TABLE 2 control sample structure

The test results are shown in Table 3.

TABLE 3 WEE1 enzyme inhibition Activity test results

Numbering of compounds	WEE1IC ₅₀ ,nM	Numbering of compounds	WEE1IC ₅₀ ,nM
				Control 1	2.57	I-1	2.63
I-2	1.83	I-3	1.84
				I-5	1.86	I-11	9.75
I-15	7.74	I-16	6.09
				I-19	3.12	I-25	3.38
I-32	3.68	I-33	6.29
				I-34	4.58	I-35	1.68
I-37	1.59	I-38	2.33
				I-40	7.99	I-41	3.71
I-42	5.27	I-46	3.06
				I-47	2.73	I-53	4.95
I-54	1.90	I-55	1.52
				I-56	4.38	I-57	1.02
I-58	2.57

Effect example 2

In vivo bioavailability experiments in mice

1. Experimental animal and test article

1. Experimental animal

All of the above were supplied by Shanghai Sipulbika laboratory animals Co.

2. Test sample preparation

2.1 preparation of mother liquor

The compound powder of the invention is added with 404.6 mu l of DMSO, is completely dissolved, is prepared into 50mg/ml mother solution, and is clear in state.

2.2 formulation of dosing solutions

The compounds of the invention: the mother solution was measured in an amount of 24. Mu.l in a physiological saline solution of 0.9%: PEG400 = 8:2 was diluted to 4mL in the ratio of 0.3mg/mL to prepare a solution which was clear and used as a solution for intravenous administration. In addition, 80 μl of mother solution is precisely measured, 0.5% CMC-Na is added to 8mL and ground into a uniform suspension state, and the concentration is as follows: 0.5mg/mL as a gastric administration solution.

2. Animal experiment

Vein group: ICR mice 24, 20±2g, intravenous injection of an intravenous injection dosing solution of the compound of the present invention, dosing volume: 10ml/kg, dose: 3mg/kg, and 0.08ml of blood was collected from the ocular fundus vein of the mice 2, 5, 15, 30, 60, 90, 120, 240, 360, 480, 600, 1440 minutes before and after the administration.

Gastric lavage group: 24 ICR mice, 20.+ -.2 g, were given by gavage the inventive compound of gavage dosing solution, dosing volume: 20ml/kg, dose: 10mg/kg, and 0.08ml of blood was collected from the ocular fundus vein of the mice 5, 15, 30, 60, 90, 120, 240, 360, 480, 600, 1440 minutes before and after the administration.

Blood samples were centrifuged at 8000rpm for 5min and plasma was taken from the centrifuge tube and stored at-20℃for further use.

3. Plasma sample processing

1. Standard curve preparation

Concentration range of standard curve working solution: 60 20,6,2,0.6,0.2,0.1,0.04,0.02. Mu.g/mL.

Taking 47.5 mu L of blank mouse plasma, adding 2.5 mu L of standard curve working solution, preparing a series of concentration samples of 3,1,0.3,0.1,0.0.3,0.01,0.005,0.002,0.001 mu g/mL, mixing uniformly by vortex, adding 300 mu L of acetonitrile (Propranolol, 25 ng/mL) containing an internal standard, precipitating protein, oscillating by vortex for 10min,6000g, centrifuging for 10min at 4 ℃, and taking supernatant for injection in a 96-well plate.

QC sample processing

Concentration range of QC working solution: low:0.06 μg/mL; middle:1.6 μg/mL; high:48 μg/mL.

Taking 47.5 mu L of blank mouse plasma, adding 2.5 mu L of QC working solution, preparing a series of concentration samples of 2.4,0.08,0.003 mu g/mL, mixing uniformly by vortex, adding 300 mu L of acetonitrile (Propranolol, 25 ng/mL) containing an internal standard to precipitate protein, oscillating by vortex for 10min,6000g, centrifuging at 4 ℃ for 10min, and taking supernatant for injection in a 96-well plate.

3. Plasma sample processing

Plasma samples 50. Mu.L were added 300. Mu.L of acetonitrile (Propranolol, 25 ng/mL) containing an internal standard to precipitate proteins, vortexed for 10min,6000g,4℃and centrifuged for 10min, the supernatant taken 1 hour before intravenous injection was diluted 10-fold with acetonitrile containing an internal standard, the remaining supernatant was undiluted, again 6000g,4℃and centrifuged for 10min, and the supernatant was taken in 96-well plates for injection.

4. Results of in vivo bioavailability experiments in mice

1. Test parameters

Dosage Dose; peak concentration C _max The method comprises the steps of carrying out a first treatment on the surface of the Peak time T _max ；AUC _last Area under the time-of-drug curve from 0 to time t; half-life T _1/2 The method comprises the steps of carrying out a first treatment on the surface of the Average residence time MRT; clearance rate Cl; apparent distribution volume V _z The method comprises the steps of carrying out a first treatment on the surface of the Steady state distribution volume V _ss The method comprises the steps of carrying out a first treatment on the surface of the Absolute bioavailability F.

2. Data on mouse pharmacokinetics

The drug parameters in mice following intravenous or intragastric administration of the compounds are shown in table 4 below.

In vivo drug substitution parameters of the compounds of Table 4 in mice

Conclusion: the compounds of the invention can significantly improve the pharmacokinetics of mice.

Claims

1. Pyrazolopyrimidine compound shown as formula A or pharmaceutically acceptable salt thereof;

x is a single bond or CH ₂ ；

Y is N or CH;

z is a single bond or CH ₂ ；

R ¹ Is H, -C (=O) R ^1-1 、-C(＝NR ^1-2 )R ^1-3 、Optionally by 1, 2 or 3R ^1-5 Substituted C ₁ ～C ₇ Alkyl, optionally substituted by 1C ₃ -C ₆ Cycloalkyl-substituted C ₁ ～C ₇ Alkyl, optionally substituted with 1, 2 or 3R ^1-5 Substituted C ₂ ～C ₇ Alkenyl, optionally substituted with 1, 2 or 3R ^1-5 Substituted C ₃ -C ₆ Cycloalkyl or optionally substituted with 1, 2 or 3R' s ^1-5 A substituted 3-7 membered ring monocyclic heterocycloalkyl;

R ^1-1 is hydrogen, cyano, -NR ^1-1-1 R ^1-1-2 、-OR ^1-1-3 Optionally by 1, 2 or 3R ^1-1-4 Substituted C ₁ ～C ₇ Alkyl, optionally substituted with 1, 2 or 3R ^1-1-4 Substituted C ₂ ～C ₇ Alkenyl, optionally substituted with 1, 2 or 3R ^1-1-4 Substituted C ₂ ～C ₇ Alkynyl or optionally substituted with 1, 2 or 3R ^1-1-4 Substituted C ₃ -C ₆ Cycloalkyl;

R ^1-1-1 and R is ^1-1-2 Independently hydrogen or C ₁ ～C ₇ An alkyl group;

R ^1-1-3 is hydrogen or C ₁ ～C ₇ An alkyl group;

each R is ^1-1-4 Independently halogen, -OH, amino, mercapto or C ₁ ～C ₇ An alkyl group; r is R ^1-2 Is hydrogen;

R ^1-3 is-NR ^1-3-1 R ^1-3-2 ；R ^1-3-1 And R is ^1-3-2 Independently hydrogen;

each R is ^1-5 Independently halogen, -OH, -SH, -CN, -NR ^1-5-1 R ^1-5-2 、-(C＝O)R ^1-5-3 、C ₁ ～C ₇ Alkyl or C ₁ ～C ₇ An alkoxy group;

R ^1-5-3 independently hydrogen, -OR ^1-5-3-1 Or C ₁ ～C ₇ An alkyl group;

R ^1-5-3-1 independently hydrogen or C ₁ ～C ₇ An alkyl group;

R ^1-6 and R is ^1-7 Independently hydrogen or C ₁ ～C ₇ An alkyl group;

R ² optionally by 1, 2 or 3R ^2-1 Substituted C ₁ ～C ₇ Alkyl, optionally substituted with 1, 2 or 3R ^2-1 Substituted C ₃ -C ₆ Cycloalkyl or optionally substituted with 1, 2 or 3R' s ^2-1 A substituted 3-7 membered ring monocyclic heterocycloalkyl;

each R is ^2-1 Independently halogen, hydroxy, amino, C ₁ ～C ₇ Alkyl or C ₁ ～C ₇ An alkoxy group; in any of the above cases, the heteroatoms in the 3-7 membered ring monocyclic heterocycloalkyl are independently selected from one or more of oxygen, sulfur, and nitrogen; the number of heteroatoms is independently 1, 2, 3 or 4;

The pyrazolopyrimidine compound shown in the formula A is not

2. The pyrazolopyrimidine compound of formula a or a pharmaceutically acceptable salt thereof according to claim 1, wherein the pyrazolopyrimidine compound of formula a or a pharmaceutically acceptable salt thereof is a pyrazolopyrimidine compound of formula I or a pharmaceutically acceptable salt thereof;

R ¹ is H, -C (=O) R ^1-1 、-C(＝NR ^1-2 )R ^1-3 Optionally by 1, 2 or 3R ^1-5 Substituted C ₁ ～C ₇ Alkyl, optionally substituted by 1C ₃ -C ₆ Cycloalkyl-substituted C ₁ ～C ₇ Alkyl, optionally substituted with 1, 2 or 3R ^1-5 Substituted C ₂ ～C ₇ Alkenyl, optionally substituted with 1, 2 or 3R ^1-5 Substituted C ₃ -C ₆ Cycloalkyl or optionally substituted with 1, 2 or 3R' s ^1-5 A substituted 3-7 membered ring monocyclic heterocycloalkyl;

R ^1-1-3 is hydrogen or C ₁ ～C ₇ An alkyl group;

each R is ^1-1-4 Independently halogen, -OH, amino, mercapto, or C ₁ ～C ₇ An alkyl group; r is R ^1-2 Is hydrogen;

R ^1-3 is-NR ^1-3-1 R ^1-3-2 ；

R ^1-3-1 And R is ^1-3-2 Independently hydrogen;

Each R is ^1-5 Independently halogen, -OH, -SH, -CN, -NR ^1-5-1 R ^1-5-2 、-(C＝O)R ^1-5-3 、C ₁ ～C ₇ Alkyl or C ₁ ～C ₇ An alkoxy group; r is R ^1-5-1 And R is ^1-5-2 Independently hydrogen or C ₁ ～C ₇ An alkyl group;

R ^1-5-3 independently hydrogen, -OR ^1-5-3-1 Or C ₁ ～C ₇ An alkyl group;

R ^1-5-3-1 independently hydrogen or C ₁ ～C ₇ An alkyl group; r is R ² Optionally by 1, 2 or 3R ^2-1 Substituted C ₁ ～C ₇ Alkyl, optionally substituted with 1, 2 or 3R ^2-1 Substituted C ₃ -C ₆ Cycloalkyl or optionally substituted with 1, 2 or 3R' s ^2-1 A substituted 3-7 membered ring monocyclic heterocycloalkyl;

each R is ^2-1 Independently halogen, hydroxy, amino, C ₁ ～C ₇ Alkyl or C ₁ ～C ₇ An alkoxy group; in any of the above cases, the heteroatoms in the 3-7 membered ring monocyclic heterocycloalkyl are independently selected from one or more of oxygen, sulfur, and nitrogen; the number of heteroatoms is independently 1, 2, 3 or 4.

3. The pyrazolopyrimidine compound of formula a or a pharmaceutically acceptable salt thereof as claimed in claim 1, whereinIs->

And/or the number of the groups of groups,is->

And/or when R ¹ Optionally by 1, 2 or 3R ^1-5 Substituted C ₁ ～C ₇ In the case of alkyl, said C ₁ ～C ₇ Alkyl is C ₁ ～C ₃ An alkyl group;

and/or when R ^1-5 When independently halogen, the halogen is fluorine, chlorine, bromine or iodine;

and/or when R ¹ Is 1R ^1-5 Substituted C ₁ ～C ₇ Alkyl, R ^1-5 When independently CN, said "is substituted with 1R ^1-5 Substituted C ₁ ～C ₇ Alkyl "is 2-cyanoethyl;

and/or when R ^1-5-3-1 Is C ₁ ～C ₇ In the case of alkyl, said C ₁ ～C ₇ Alkyl is C ₁ ～C ₃ An alkyl group;

and/or when R ¹ Optionally by 1, 2 or 3R ^1-5 When substituted 3-7 membered ring monocyclic heterocycloalkyl, R is removed ^1-5 In addition, the heteroatoms in the 3-7 membered ring monocyclic heterocycloalkyl are not substituted;

and/or when R ¹ Optionally by 1, 2 or 3R ^1-5 In the case of a substituted 3-7 membered monocyclic heterocycloalkyl, the methylene group in said 3-7 membered monocyclic heterocycloalkyl is not replaced;

and/or when R ^1-1 Optionally by 1, 2 or 3R ^1-1-4 Substituted C ₁ ～C ₇ In the case of alkyl, said C ₁ ～C ₇ Alkyl is C ₁ ～C ₃ An alkyl group;

and/or when R ^1-6 Is C ₁ ～C ₇ In the case of alkyl, said C ₁ ～C ₇ Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl;

and/or when R ^1-7 Is C ₁ ～C ₇ In the case of alkyl, said C ₁ ～C ₇ Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl;

and/or when R ² Optionally by 1, 2 or 3R ^2-1 Substituted C ₁ ～C ₇ In the case of alkyl, said C ₁ ～C ₇ Alkyl isC ₁ ～C ₄ An alkyl group;

and/or when R ² Optionally by 1, 2 or 3R ^2-1 When substituted 3-7 membered ring monocyclic heterocycloalkyl, R is removed ^2-1 In addition, the heteroatoms in the 3-7 membered ring monocyclic heterocycloalkyl are not substituted;

and/or when R ² Optionally by 1, 2 or 3R ^2-1 In the case of a substituted 3-7 membered monocyclic heterocycloalkyl, the methylene group in said 3-7 membered monocyclic heterocycloalkyl is not replaced;

and/or atoms in the pyrazolopyrimidine compound shown as the formula I or pharmaceutically acceptable salt thereof exist in the form of natural abundance.

4. The pyrazolopyrimidine compound of formula a or a pharmaceutically acceptable salt thereof as claimed in claim 3, whereinIs->Said->Is that

And/or the number of the groups of groups,is->

And/or when R ¹ Optionally by 1, 2 or 3R ^1-5 Substituted C ₁ ～C ₇ In the case of alkyl, said C ₁ ～C ₇ Alkyl groupMethyl, ethyl, n-propyl or isopropyl;

and/or when R ^1-5 When independently halogen, the halogen is fluorine;

when R is ¹ Is optionally substituted with 1C ₃ -C ₆ Cycloalkyl-substituted C ₁ ～C ₇ In the case of alkyl groups, said radicals are optionally substituted by 1C ₃ -C ₆ Cycloalkyl-substituted C ₁ ～C ₇ C in alkyl ₃ -C ₆ Cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl;

and/or when R ^1-5-3-1 Is C ₁ ～C ₇ In the case of alkyl, said C ₁ ～C ₇ Alkyl is methyl, ethyl, n-propyl or isopropyl;

and/or when R ¹ Optionally by 1, 2 or 3R ^1-5 Substituted C ₃ -C ₆ In the case of cycloalkyl, said C ₃ -C ₆ Cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl;

and/or when R ¹ Optionally by 1, 2 or 3R ^1-5 In the case of a substituted 3-7 membered monocyclic heterocycloalkyl, the 3-7 membered monocyclic heterocycloalkyl is

And/or when R ^1-1 Optionally by 1, 2 or 3R ^1-1-4 Substituted C ₁ ～C ₇ In the case of alkyl, said C ₁ ～C ₇ Alkyl is methyl, ethyl, n-propyl or isopropyl;

and/or when R ^1-1 Optionally by 1, 2 or 3R ^1-1-4 Substituted C ₃ -C ₆ In the case of cycloalkyl, said C ₃ -C ₆ Cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl;

and/or when R ^1-6 Is C ₁ ～C ₇ In the case of alkyl, said C ₁ ～C ₇ Alkyl is methyl or ethyl;

and/or when R ^1-7 Is C ₁ ～C ₇ In the case of alkyl, said C ₁ ～C ₇ Alkyl is methyl or ethyl;

and/or when R ² Optionally by 1, 2 or 3R ^2-1 Substituted C ₁ ～C ₇ In the case of alkyl, said C ₁ ～C ₇ Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl;

and/or when R ² Optionally by 1, 2 or 3R ^2-1 In the case of a substituted 3-7 membered monocyclic heterocycloalkyl, the 3-7 membered monocyclic heterocycloalkyl is oxetan-3-yl.

5. The pyrazolopyrimidine compound of formula A or a pharmaceutically acceptable salt thereof according to claim 4, Is->

And/or when R ¹ Is 1, 2 or 3R ^1-5 Substituted C ₁ ～C ₇ Alkyl, optionally substituted by 1C ₃ -C ₆ Cycloalkyl-substituted C ₁ ～C ₇ Alkyl, R ^1-5 Independently halogen, said "is substituted with 1, 2 or 3R ^1-5 Substituted C ₁ ～C ₇ Alkyl "is 2, 2-difluoroethyl or 3, 3-trifluoropropyl;

and/or when R ¹ Is 1R ^1-5 Substituted C ₃ -C ₆ Cycloalkyl, R ^1-5 Is independently-NR ^1-5-1 R ^1-5-2 When said is 1R ^1-5 Substituted C ₃ -C ₆ Cycloalkyl "is 3-aminocyclobutyl;

and/or when R ¹ Is optionally substituted with 1R ^1-5 Substituted 3-7 membered ring monocyclic heterocycloalkyl, R ^1-5 Is C ₁ ～C ₇ In the case of alkyl radicals, the radicals "optionally substituted by 1R ^1-5 A substituted 3-7 membered ring monocyclic heterocycloalkyl "is

And/or when R ^1-1 Is optionally substituted with 1R ^1-1-4 Substituted C ₁ ～C ₇ Alkyl, R ^1-1-4 In the case of-OH, the said "optionally substituted with 1R ¹ ^-1-4 Substituted C ₁ ～C ₇ Alkyl "is hydroxymethyl;

and/or when R ² Is 1R ^2-1 Substituted C ₁ ～C ₇ Alkyl, R ^2-1 In the case of hydroxyl, said quilt has 1R ^2-1 Substituted C ₁ ～C ₇ Alkyl is

And/or when R ² Is 1R ^2-1 Substituted 3-7 membered ring monocyclic heterocycloalkyl, R ^2-1 In the case of halogen or hydroxy, said quilt has 1R ^2-1 The substituted 3-7 membered ring monocyclic heterocycloalkyl is

6. Pyrazolopyrimidine compound of the formula a or a pharmaceutically acceptable salt thereof as claimed in claim 1 or 2, wherein R ¹ Is H, -C (=O) R ^1-1 、-C(＝NR ^1-2 )R ^1-3 Optionally by 1, 2 or 3R ^1-5 Substituted C ₁ ～C ₇ Alkyl, optionally substituted by 1C ₃ -C ₆ Cycloalkyl-substituted C ₁ ～C ₇ Alkyl, optionally substituted with 1, 2 or 3R ^1-5 Substituted C ₃ -C ₆ Cycloalkyl, or optionally substituted with 1, 2 or 3R ^1-5 Substituted 3-7 membered ringsA monocyclic heterocycloalkyl group;

and/or R ^1-1 is-NR ^1-1-1 R ^1-1-2 、-OR ^1-1-3 Cyano, optionally substituted with 1R ^1-1-4 Substituted C ₁ ～C ₇ Alkyl, optionally substituted with 1R ^1-1-4 Substituted C ₂ ～C ₇ Alkynyl or optionally substituted with 1R ^1-1-4 Substituted C ₃ -C ₆ Cycloalkyl; r is R ^1-1-1 And R is ^1-1-2 Is hydrogen or C ₁ ～C ₇ An alkyl group; r is R ^1-1-3 Is hydrogen or C ₁ ～C ₇ An alkyl group; r is R ^1-1-4 Halogen, OH or C ₁ ～C ₇ An alkyl group;

and/or each R ^1-5 Independently halogen, -CN, -NR ^1-5-1 R ^1-5-2 、-(C＝O)R ^1-5-3 Or C ₁ ～C ₇ An alkyl group;

R ^1-5-1 and R is ^1-5-2 Independently hydrogen or C ₁ ～C ₇ An alkyl group; r is R ^1-5-3 Independently hydrogen, -OR ^1-5-3-1 Or C ₁ ～C ₇ An alkyl group; r is R ¹ ^-5-3-1 Is C ₁ ～C ₇ An alkyl group;

and/or R ² Is 1R ^2-1 Substituted C ₁ ～C ₇ Alkyl or by 1R ^2-1 A substituted 3-7 membered ring monocyclic heterocycloalkyl; each R is ^2-1 Independently a hydroxyl group or a halogen.

7. Pyrazolopyrimidine compound of the formula A or a pharmaceutically acceptable salt thereof as claimed in claim 1, wherein R ¹ Is H, -C (=O) R ^1-1 、Optionally by 1, 2 or 3R ^1-5 Substituted C ₁ ～C ₇ Alkyl, optionally substituted by 1C ₃ -C ₆ Cycloalkyl-substituted C ₁ ～C ₇ Alkyl, optionally substituted with 1, 2 or 3R ^1-5 Substituted C ₃ -C ₆ Cycloalkyl, or, optionally, 1, 2 or 3 R ^1-5 A substituted 3-7 membered ring monocyclic heterocycloalkyl;

and/or R ^1-1 Is optionally substituted with 1R ^1-1-4 Substituted C ₁ ～C ₇ Alkyl or optionally substituted with 1R ^1-1-4 Substituted C ₃ -C ₆ Cycloalkyl; r is R ^1-1-4 is-OH;

and/or each R ^1-5 Independently halogen, -CN, -NR ^1-5-1 R ^1-5-2 、-(C＝O)R ^1-5-3 Or C ₁ ～C ₇ An alkyl group; r is R ^1-5-1 And R is ¹ ^-5-2 Is hydrogen; r is R ^1-5-3 Independently is-OR ^1-5-3-1 ；R ^1-5-3-1 Independently C ₁ ～C ₇ An alkyl group;

and/or R ² Is 1R ^2-1 Substituted C ₁ ～C ₇ Alkyl or is substituted by 1R ^2-1 A substituted 3-7 membered ring monocyclic heterocycloalkyl; each R is ^2-1 Is hydroxyl;

and/or X is a single bond or CH ₂ The method comprises the steps of carrying out a first treatment on the surface of the Y is C; z is a single bond or CH ₂ 。

8. The pyrazolopyrimidine compound of formula a or a pharmaceutically acceptable salt thereof as claimed in claim 7, wherein R ¹ Is H, -C (=O) R ^1-1 Optionally by 1, 2 or 3R ^1-5 Substituted C ₁ ～C ₇ Alkyl, optionally substituted by 1C ₃ -C ₆ Cycloalkyl-substituted C ₁ ～C ₇ Alkyl, optionally substituted with 1, 2 or 3R ^1-5 Substituted C ₃ -C ₆ Cycloalkyl, or, optionally, is substituted with 1, 2 or 3R ^1-5 A substituted 3-7 membered ring monocyclic heterocycloalkyl.

9. The pyrazolopyrimidine compound of formula a or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein the pyrazolopyrimidine compound of formula a is defined as follows:

scheme (1):

R ¹ is H, -C (=O) R ^1-1 、-C(＝NR ^1-2 )R ^1-3 Optionally by 1, 2 or 3R ^1-5 Substituted C ₁ ～C ₇ Alkyl, optionally substituted by 1C ₃ -C ₆ Cycloalkyl-substituted C ₁ ～C ₇ Alkyl, optionally substituted with 1, 2 or 3R ^1-5 Substituted C ₃ -C ₆ Cycloalkyl or optionally substituted with 1, 2 or 3R' s ^1-5 A substituted 3-7 membered ring monocyclic heterocycloalkyl;

R ^1-1 is-NR ^1-1-1 R ^1-1-2 、-OR ^1-1-3 Cyano, optionally substituted with 1R ^1-1-4 Substituted C ₁ ～C ₇ Alkyl, optionally substituted with 1R ^1-1-4 Substituted C ₂ ～C ₇ Alkynyl, optionally substituted with 1R ^1-1-4 Substituted C ₃ -C ₆ Cycloalkyl; r is R ^1-1-1 And R is ^1-1-2 Is hydrogen or C ₁ ～C ₇ An alkyl group; r is R ¹ ^-1-3 Is hydrogen or C ₁ ～C ₇ An alkyl group; each R is ^1-1-4 Halogen, OH or C ₁ ～C ₇ An alkyl group;

R ^1-2 is hydrogen;

R ^1-5 independently halogen, -CN, -NR ^1-5-1 R ^1-5-2 、-(C＝O)R ^1-5-3 Or C ₁ ～C ₇ An alkyl group; r is R ^1-5-1 And R is ^1-5-2 Independently hydrogen or C ₁ ～C ₇ An alkyl group;

R ² is 1R ^2-1 Substituted C ₁ ～C ₇ Alkyl or by 1R ^2-1 A substituted 3-7 membered ring monocyclic heterocycloalkyl; each R is ^2-1 Independently a hydroxyl group or a halogen.

10. The pyrazolopyrimidine compound of formula a or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein the pyrazolopyrimidine compound of formula a is defined as follows:

scheme (2):

the pyrazolopyrimidine compound shown in the formula A is pyrazolopyrimidine compound shown in the formula I;

R ¹ Is H, -C (=O) R ^1-1 、-C(＝NR ^1-2 )R ^1-3 Optionally by 1, 2 or 3R ^1-5 Substituted C ₁ ～C ₇ Alkyl, optionally substituted by 1C ₃ -C ₆ Cycloalkyl-substituted C ₁ ～C ₇ Alkyl, optionally substituted with 1, 2 or 3R ^1-5 Substituted C ₃ -C ₆ Cycloalkyl or optionally substituted with 1, 2 or 3R' s ^1-5 A substituted 3-7 membered ring monocyclic heterocycloalkyl; r is R ^1-1 is-NR ^1-1-1 R ^1-1-2 、-OR ^1-1-3 Cyano, optionally substituted with 1R ^1-1-4 Substituted C ₁ ～C ₇ Alkyl, optionally substituted with 1R ^1-1-4 Substituted C ₂ ～C ₇ Alkynyl or optionally substituted with 1R ^1-1-4 Substituted C ₃ -C ₆ Cycloalkyl; r is R ^1-1-1 And R is ^1-1-2 Is hydrogen or C ₁ ～C ₇ An alkyl group; r is R ^1-1-3 Is hydrogen or C ₁ ～C ₇ An alkyl group; each R is ^1-1-4 Halogen, OH or C ₁ ～C ₇ An alkyl group;

R ^1-2 is hydrogen;

each R is ^1-5 Independently halogen, -CN, -NR ^1-5-1 R ^1-5-2 、-(C＝O)R ^1-5-3 Or C ₁ ～C ₇ An alkyl group; r is R ^1-5-1 And R is ^1-5-2 Independently hydrogen or C ₁ ～C ₇ An alkyl group;

11. The pyrazolopyrimidine compound of formula a or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein the pyrazolopyrimidine compound of formula a is defined as follows:

scheme (3):

R ¹ is H, -C (=O) R ^1-1 、Optionally by 1, 2 or 3R ^1-5 Substituted C ₁ ～C ₇ Alkyl, optionally substituted by 1C ₃ -C ₆ Cycloalkyl-substituted C ₁ ～C ₇ Alkyl, optionally substituted with 1, 2 or 3R ^1-5 Substituted C ₃ -C ₆ Cycloalkyl or optionally substituted with 1, 2 or 3R' s ^1-5 A substituted 3-7 membered ring monocyclic heterocycloalkyl; r is R ^1-1 Is optionally substituted with 1R ^1-1-4 Substituted C ₁ ～C ₇ Alkyl or optionally substituted with 1R ^1-1-4 Substituted C ₃ -C ₆ Cycloalkyl; each R is ^1-1-4 is-OH;

each R is ^1-5 Independently halogen, -CN, -NR ^1-5-1 R ^1-5-2 、-(C＝O)R ^1-5-3 Or C ₁ ～C ₇ An alkyl group; r is R ^1-5-1 And R is ^1-5-2 Is hydrogen;

12. The pyrazolopyrimidine compound of formula a or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein the pyrazolopyrimidine compound of formula a is defined as follows:

scheme (4):

R ¹ is H, -C (=O) R ^1-1 Optionally by 1, 2 or 3R ^1-5 Substituted C ₁ ～C ₇ Alkyl, optionally substituted by 1C ₃ -C ₆ Cycloalkyl-substituted C ₁ ～C ₇ Alkyl, optionally substituted with 1, 2 or 3R ^1-5 Substituted C ₃ -C ₆ Cycloalkyl or optionally substituted with 1, 2 or 3R' s ^1-5 A substituted 3-7 membered ring monocyclic heterocycloalkyl;

R ^1-1 is optionally substituted with 1R ^1-1-4 Substituted C ₁ ～C ₇ Alkyl or optionally substituted with 1R ^1-1-4 Substituted C ₃ -C ₆ Cycloalkyl; r is R ^1-1-4 is-OH;

R ² is 1R ^2-1 Substituted: c (C) ₁ ～C ₇ Alkyl or a 3-7 membered ring monocyclic heterocycloalkyl; r is R ^2-1 Independently a hydroxyl group or a halogen.

13. The pyrazolopyrimidine compound of formula a or a pharmaceutically acceptable salt thereof according to claim 1, wherein the pyrazolopyrimidine compound of formula a is defined as follows:

scheme (5):

the pyrazolopyrimidine compound shown in the formula A is a pyrazolopyrimidine compound shown in the formula A-1;

R ¹ is H, -C (=O) R ^1-1 、Optionally by 1, 2 or 3R ^1-5 Substituted C ₁ ～C ₇ Alkyl, optionally substituted by 1C ₃ -C ₆ Cycloalkyl-substituted C ₁ ～C ₇ Alkyl, optionally substituted with 1, 2 or 3R ^1-5 Substituted C ₃ -C ₆ Cycloalkyl or optionally substituted with 1, 2 or 3R' s ^1-5 A substituted 3-7 membered ring monocyclic heterocycloalkyl;

R ^1-1 is optionally substituted with 1R ^1-1-4 Substituted C ₁ ～C ₇ Alkyl or optionally substituted with 1R ^1-1-4 Substituted C ₃ -C ₆ Cycloalkyl; each R is ¹ ^-1-4 is-OH;

R ^1-6 and R is ^1-7 Independently hydrogen or C ₁ ～C ₇ An alkyl group;

R ² is 1R ^2-1 Substituted C ₁ ～C ₇ Alkyl or is substituted by 1R ^2-1 A substituted 3-7 membered ring monocyclic heterocycloalkyl; each R is ^2-1 Is hydroxyl.

14. The pyrazolopyrimidine compound of formula a or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein the pyrazolopyrimidine compound of formula a is defined as follows:

scheme (6):

the pyrazolopyrimidine compound shown in the formula A is a pyrazolopyrimidine compound shown in the formula I-1;

R ¹ is H, -C (=O) R ^1-1 Optionally by 1, 2 or 3R ^1-5 Substituted C ₁ ～C ₇ Alkyl, optionally substituted by 1C ₃ -C ₆ Cycloalkyl-substituted C ₁ ～C ₇ Alkyl, optionally substituted with 1, 2 or 3R ^1-5 Substituted C ₃ -C ₆ Cycloalkyl, or, optionally, is substituted with 1, 2 or 3R ^1-5 A substituted 3-7 membered ring monocyclic heterocycloalkyl;

R ^1-1 is optionally substituted with 1R ^1-1-4 Substituted C ₁ ～C ₇ Alkyl or optionally substituted with 1R ^1-1-4 Substituted C ₃ -C ₆ Cycloalkyl; each R is ^1-1-4 is-OH;

R ² is 1 to be covered byR ^2-1 Substituted C ₁ ～C ₇ Alkyl or by 1R ^2-1 A substituted 3-7 membered ring monocyclic heterocycloalkyl; each R is ^2-1 Is hydroxyl.

15. The pyrazolopyrimidine compound of formula a or a pharmaceutically acceptable salt thereof according to claim 1, wherein the pyrazolopyrimidine compound of formula a is defined as follows:

scheme (7):

R ¹ Is that

R ^1-6 And R is ^1-7 Independently C ₁ ～C ₇ An alkyl group;

R ² optionally by 1, 2 or 3R ^2-1 Substituted C ₁ ～C ₇ Alkyl or optionally substituted with 1, 2 or 3R ^2-1 A substituted 3-7 membered ring monocyclic heterocycloalkyl; each R is ^2-1 Is hydroxyl.

16. The pyrazolopyrimidine compound of formula a or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein the pyrazolopyrimidine compound of formula a has any one of the following structures:

17. a process for the preparation of pyrazolopyrimidine compound of formula a as claimed in any one of claims 1 to 16, which is any one of the following:

wherein R is ¹ 、R ^1-1 、R ² X, Y and Z are as defined in any one of claims 1 to 16, PG being an amino protecting group.

18. The method for preparing pyrazolopyrimidine compound of formula a according to claim 17, wherein the pyrazolopyrimidine compound of formula a is a pyrazolopyrimidine compound of formula I, which is any one of the following methods:

19. A compound of formula 2A:

wherein,

R ¹ is 1, 2 or 3R ^1-5 Substituted C ₁ ～C ₇ An alkyl group; said R is ^1-5 is-CN.

20. A compound characterized in that it is the following compound:

21. use of substance X in the preparation of a kinase inhibitor;

the substance X is pyrazolopyrimidine compound shown in the formula A or pharmaceutically acceptable salt thereof as claimed in any one of claims 1-16.

22. The application of a substance X in the preparation of medicines; the medicament is used for treating and/or preventing diseases related to WEE1 kinase or treating and/or preventing cancers;

23. A pharmaceutical composition comprising substance X and a pharmaceutical adjuvant;

24. A combination comprising a substance X and an anticancer drug,

25. Use of a combination according to claim 24 for the manufacture of a medicament for the prevention and/or treatment of cancer.

26. A pharmaceutical composition comprising the combination of claim 24 and a pharmaceutically acceptable adjuvant.

27. A combination kit comprising pharmaceutical composition a and pharmaceutical composition B;

the medicine composition A comprises a substance X and pharmaceutic adjuvant;

the medicine composition B comprises an anticancer medicine and a pharmaceutic adjuvant;