WO2022033552A1

WO2022033552A1 - Cdk kinase inhibitor, preparation method therefor, pharmaceutical composition, and application

Info

Publication number: WO2022033552A1
Application number: PCT/CN2021/112286
Authority: WO
Inventors: 杨磊
Original assignee: 隆泰申医药科技(南京) 有限公司
Priority date: 2020-08-12
Filing date: 2021-08-12
Publication date: 2022-02-17
Also published as: CN114075205A

Abstract

Disclosed in the present invention are a CDK kinase inhibitor, a preparation method therefor, a pharmaceutical composition, and an application. The present invention provides a compound as represented by formula I-a, a cis-trans isomer thereof, a pharmaceutically acceptable salt thereof, a solvate of the pharmaceutically acceptable salt thereof, a tautomer thereof, or a polymorph thereof. The compound of the present invention has good CDK7 inhibitory activity, cell activity, and tumor growth inhibitory activity.

Description

CDK kinase inhibitor, its preparation method, pharmaceutical composition and application

This application claims the priority of Chinese patent application 2020108066604 with an application date of 2020/8/12. This application cites the full text of the above Chinese patent application.

technical field

The present invention relates to a CDK kinase inhibitor, its preparation method, pharmaceutical composition and application.

Background technique

Cyclin-dependent kinases (CDKs) are relatively small proteins with a molecular weight between 34 and 40 kilodaltons that contain essentially only the kinase domain. CDKs bind regulatory proteins called cyclins. In the absence of cyclin, CDKs have little kinase activity; only the cyclin-CDK complex is an active kinase. CDKs phosphorylate the serine/threonine residues of their substrates and therefore belong to the class of serine/threonine kinases (David O. Morgan, The Cell Cycle: Principles of Control. London: New Science Press ), 1st edition, (2007)).

Members of the cyclin-dependent kinase (CDK) family play key regulatory roles in cell proliferation. There are currently 20 known mammalian CDKs. Although CDK7-13 and CDK18 were associated with transcription, only CDK1, CDK2, CDK4 and CDK6 showed a clear association with the cell cycle. As a unique CDKs among mammalian CDKs, CDK7 enhances kinase activity and regulates cell cycle and transcription. In the cytosol, CDK7 exists as a heterotrimeric complex and is thought to function as a CDK1/2-activating kinase (CAK), which is required for fully catalyzed CDK activity and cell cycle progression for CDK1/2 Conserved residues are phosphorylated (Desai et al., Mol. Cell Biol. 15, 345-350 (1995)).

CDK7 in the CDK family has been shown to be closely related to the occurrence and development of various malignant tumors. CDK7 inhibitors may be used for the treatment of various malignant tumors.

SUMMARY OF THE INVENTION

The technical problem to be solved by the present invention is to provide a CDK kinase inhibitor different from the prior art, its preparation method, pharmaceutical composition and application in order to overcome the lack of existing CDK7 kinase inhibitors in the prior art. The compounds of the present invention have better inhibitory activity as CDK kinase inhibitors.

The present invention mainly solves the above technical problems through the following technical solutions.

The present invention provides a compound shown in formula Ia, its cis-trans isomer, its pharmaceutically acceptable salt, its solvate, its pharmaceutically acceptable salt solvate, its tautomerism a form or a polymorph thereof,

Wherein, R ¹ and R ² are independently C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl substituted by one or more halogens (when there are multiple halogens, the halogens are the same or different), C ₃ -C ₆ cycloalkyl or -NR ^1-1 R ^1-2 ;

R ^1-1 and R ^1-2 are independently H or 5-8-membered heterocycloalkyl, and the heteroatom in the 5-8-membered heterocycloalkyl is one or more of N, S and O , the number is 1, 2 or 3 (when the number of heteroatoms is 2 or 3, the types of heteroatoms may be the same or different);

X is N or CR ³ , R ³ is hydrogen or halogen;

Y is N or O;

R ⁴ is H or C ₁ -C ₆ alkyl;

n is 1 or 2;

n ^a is 1 or 2;

Z is N or CH;

L is NH or absent (ie, Z is directly attached to the carbon group);

R ⁵ is H or halogen;

R ⁶ and R ⁷ are independently H, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl substituted by one or more R ^6-1 (when R ^6-1 is more than one, the R ^6-1 same or different), 5-8-membered heterocycloalkyl or 5-8-membered heterocycloalkyl substituted by one or more R ^6-2 (when R ^6-2 is more than one, the R ^6-2 are the same or different), the heteroatoms in the 5-8-membered heterocycloalkyl, and, one or more R ^6-2 -substituted 5-8-membered heterocycloalkyl are independently N, S and one or more of O, the number of which is 1, 2 or 3 (when the number of heteroatoms is 2 or 3, the types of heteroatoms may be the same or different);

R ^6-1 is -NR ^6-1-1 R ^6-1-2 ;

R ^6-1-1 and R ^6-1-2 are independently H or C ₁ -C ₆ alkyl,

Alternatively, R ^6-1-1 and R ^6-1-2 together with the N to which they are attached form a 5-8 membered heterocycloalkyl or, one or more C ₁ -C ₆ alkyl substituted 5-8 membered heterocycle Alkyl; the heteroatoms in the 5-8-membered heterocycloalkyl group and the 5-8-membered heterocycloalkyl group substituted by one or more C ₁ -C ₆ alkyl groups are independently one of N, S and O One or more, the number is 1, 2 or 3 (when the number of heteroatoms is 2 or 3, the types of heteroatoms may be the same or different);

R ^6-2 is C ₁ -C ₆ alkyl;

When n is 2, X is N;

The carbon atoms marked with * are chiral carbon in R configuration, chiral carbon in S configuration or achiral carbon.

In one embodiment of the present invention, when R ¹ is a C ₁ -C ₆ alkyl group, the C ₁ -C ₆ alkyl group is a C ₁ -C ₄ alkyl group, preferably methyl, ethyl, n-propyl , isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, further preferably isopropyl or tert-butyl, eg isopropyl.

In one embodiment of the present invention, when R ¹ is C ₁ -C ₆ alkyl substituted by one or more halogens, the halogen is fluorine, chlorine, bromine or iodine; the C ₁ -C ₆ alkyl Preferably it is a C ₁ -C ₄ alkyl group, more preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl.

In one embodiment of the present invention, when R ¹ is a C ₃ -C ₆ cycloalkyl group, the C ₃ -C ₆ cycloalkyl group is a cyclopropyl group, a cyclobutyl group, a cyclopentyl group or a cyclohexyl group, such as Cyclopropanyl.

In an embodiment of the present invention, when R ² is a C ₁ -C ₆ alkyl group, the C ₁ -C ₆ alkyl group is a C ₁ -C ₄ alkyl group, preferably methyl, ethyl, n-propyl group, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, more preferably methyl.

In one embodiment of the present invention, when R ² is C ₁ -C ₆ alkyl substituted by one or more halogens, the halogen is fluorine, chlorine, bromine or iodine; the C ₁ -C ₆ alkyl Preferably it is a C ₁ -C ₄ alkyl group, more preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl.

In one embodiment of the present invention, when R ^1-1 and R ^1-2 are independently a 5-8 membered heterocycloalkyl, the 5-8 membered heterocycloalkyl is a 5-6 membered heterocycloalkane base, the heteroatom is O, and the number is 1; preferably pyranyl, for example

In one embodiment of the present invention, when R ² is -NR ^1-1 R ^1-2 , one of R ^1-1 and R ^1-2 is H, and the other is 5-8 membered heterocycloalkyl. Described-NR ^1-1 R ^1-2 is preferably

In one embodiment of the present invention, when ^R3 is halogen, the halogen is fluorine, chlorine, bromine or iodine.

In an embodiment of the present invention, when R ⁴ is a C ₁ -C ₆ alkyl group, the C ₁ -C ₆ alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl , isobutyl, sec- or tert-butyl, eg methyl.

^In one embodiment of the present invention, when R5 is halogen, said halogen is fluorine, chlorine, bromine or iodine, such as fluorine.

In one embodiment of the present invention, when R ⁶ and R ⁷ are independently C ₁ -C ₆ alkyl, or, one or more R ^6-1 substituted C ₁ -C ₆ alkyl, the C ₁ -C ₆ alkyl group and one or more R ^6-1 substituted C ₁ -C ₆ alkyl group in which C ₁ -C ₆ alkyl group is independently methyl, ethyl, n-propyl, isopropyl , n-butyl, isobutyl, sec-butyl or tert-butyl, eg methyl.

In one embodiment of the present invention, when R ^6-1-1 and R ^6-1-2 are independently C ₁ -C ₆ alkyl, the C ₁ -C ₆ alkyl is independently methyl, ethyl , n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, eg methyl.

In one embodiment of the present invention, when R ^6-1-1 and R ^6-1-2 together with the N to which they are attached form a 5-8 membered heterocycloalkyl, the 5-8 membered heterocycloalkyl is 5-6 membered heterocycloalkyl, the heteroatoms are N and/or O, and the number is 2; preferably morpholino, for example

In one embodiment of the present invention, when R ^6-1-1 and R ^6-1-2 together with the N to which they are attached form one or more C ₁ -C ₆ alkyl substituted 5-8 membered heterocycloalkanes, The 5-8-membered heterocycloalkyl group is a 5-6-membered heterocycloalkyl group, the heteroatom is N, and the number is 1 or 2; it is preferably a piperazinyl group.

In one embodiment of the present invention, when R ^6-1-1 and R ^6-1-2 together with the N to which they are attached form one or more C ₁ -C ₆ alkyl substituted 5-8 membered heterocycloalkyl , the C ₁ -C ₆ alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, such as methyl.

In one embodiment of the present invention, when R ^6-1-1 and R ^6-1-2 together with the N to which they are attached form one or more C ₁ -C ₆ alkyl substituted 5-8 membered heterocycloalkyl , the one or more C ₁ -C ₆ alkyl substituted 5-8 membered heterocycloalkyl is methyl substituted piperazinyl, for example

In one embodiment of the present invention, when R ⁶ and R ⁷ are independently one or more R ^6-1 substituted C ₁ -C ₆ alkyl, the one or more R ^6-1 substituted C ₁ -C ₆ alkyl is

In one embodiment of the present invention, when said R ⁶ and R ⁷ are independently 5-8-membered heterocycloalkyl or 5-8-membered heterocycloalkyl substituted by one or more R ^6-2 , the The 5-8-membered heterocycloalkyl in the 5-8-membered heterocycloalkyl group and the one or more R ^6-2 substituted 5-8-membered heterocycloalkyl groups are independently 5-6-membered heterocycloalkyl groups. Cycloalkyl, the heteroatom is N and the number is 2, such as pyrrolidinyl.

In an embodiment of the present invention, when R ^6-2 is a C ₁ -C ₆ alkyl group, the C ₁ -C ₆ alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl , isobutyl, sec- or tert-butyl, eg methyl.

In one embodiment of the present invention, when said R ⁶ and R ⁷ are independently one or more R ^6-2 substituted 5-8-membered heterocycloalkyl, said one or more R ^{6- 2} -substituted 5-8 membered heterocycloalkyl is methyl-substituted pyrrolidinyl, such as

In one embodiment of the present invention,

for

In one embodiment of the present invention, R ¹ is C ₁ -C ₆ alkyl.

In one embodiment of the invention, R ¹ is C ₁ -C ₆ alkyl (eg ethyl, isopropyl or tert-butyl) or C ₃ -C ₆ cycloalkyl (eg cyclopropyl).

In one embodiment of the present invention, R ² is C ₁ -C ₆ alkyl.

In one embodiment of the invention, R ² is C ₁ -C ₆ alkyl (eg methyl) or -NR ^1-1 R ^1-2 (eg

).

In one embodiment of the present invention, ^R3 is hydrogen.

In one embodiment of the present invention, ^na is 1.

In one embodiment of the present invention, Y is O.

In one embodiment of the invention, Z is N and L is absent; or, Z is CH and L is NH.

In one embodiment of the present invention, R ⁶ and R ⁷ are independently H or, one or more R ^6-1 substituted C ₁ -C ₆ alkyl; R ^6-1 is -NR ^6- ^1-1 R ^6-1-2 ; R ^6-1-1 and R ^6-1-2 are independently C ₁ -C ₆ alkyl, or, R ^6-1-1 and R ^6-1-2 together with the N to which they are attached Forms a 5-8 membered heterocycloalkyl or a 5-8 membered heterocycloalkyl substituted with one or more _C1 - _C6 alkyl groups.

In one embodiment of the present invention, the compound represented by the formula I-a is the compound represented by the formula I-a1 or I-a2,

In one embodiment of the present invention, in the compound shown in the formula I-a1:

R ¹ and R ² are independently C ₁ -C ₆ alkyl;

Y is O;

n is 2;

n ^a is 1;

Z is N;

L means does not exist;

R ⁶ and R ⁷ are independently one or more R ^6-1 substituted C ₁ -C ₆ alkyl;

R ^6-1 is -NR ^6-1-1 R ^6-1-2 ;

R ^6-1-1 and R ^6-1-2 are independently C ₁ -C ₆ alkyl.

In an embodiment of the present invention, in the compound shown in the formula I-a2:

R ¹ is C ₁ -C ₆ alkyl;

R ² is C ₁ -C ₆ alkyl or -NR ^1-1 R ^1-2 ;

X is N or CH;

Y is O;

Z is N;

L means does not exist;

R ⁵ is H;

R ⁶ and R ⁷ are independently H, C ₁ -C ₆ alkyl, or, one or more R ^6-1 substituted C ₁ -C ₆ alkyl;

R ^6-1 is -NR ^6-1-1 R ^6-1-2 ;

R ^6-1-1 and R ^6-1-2 are independently C ₁ -C ₆ alkyl.

In one embodiment of the present invention, the compound represented by formula Ia, its cis-trans isomer, its pharmaceutically acceptable salt, its solvate, its pharmaceutically acceptable salt solvate , its tautomer, or its polymorphic form is a compound shown in formula I, its cis-trans isomer or a pharmaceutically acceptable salt thereof,

in,

R ¹ and R ² are independently C ₁ -C ₆ alkyl, or C ₁ -C ₆ alkyl substituted with one or more halogens (when there are multiple halogens, the halogens are the same or different);

X is N or CR ³ , R ³ is hydrogen or halogen;

n is 1 or 2;

The carbon atom marked with * is chiral carbon in R configuration, chiral carbon in S configuration or achiral carbon;

When n is 2, X is N.

In one embodiment of the present invention, the compound shown in formula I is the compound shown in formula I-1,

Wherein, R ¹ and R ² are independently C ₁ -C ₆ alkyl groups, or C ₁ -C ₆ alkyl groups substituted with one or more halogens; the carbon atoms marked with * are R-configuration chiral carbon, S-configuration chiral or achiral carbon.

In one embodiment of the present invention, in the compound represented by formula I-1, the carbon atom marked with * is a chiral carbon of R configuration or a chiral carbon of S configuration.

In one embodiment of the present invention, the compound shown in formula I is the compound shown in formula I-2,

Wherein, R ¹ and R ² are independently C ₁ -C ₆ alkyl, or C ₁ -C ₆ alkyl substituted by one or more halogens; X is N or CR ³ , and R ³ is hydrogen or halogen.

In an embodiment of the present invention, the compound represented by the formula I-a is

wherein R ⁸ is halogen (eg chlorine or bromine) or hydroxy; R ¹ , R ² , X, Y, R ⁴ , Z, L, ⁿ , R ⁵ , na , R ⁶ , R ⁷ and those marked with * The carbon atoms are as described above.

In the condensation reaction, the solvent may be a conventional solvent for this type of reaction in the field, and in the present invention, it is particularly preferably a nitrile solvent (eg, acetonitrile).

In the condensation reaction, the molar concentration of the compound represented by the formula II-a in the solvent can be the conventional molar concentration of this type of reaction in the field, and the present invention is particularly preferably 0.01-0.5 mol/L , more preferably 0.05 to 0.15 mol/L (for example, 0.09 mol/L).

In the condensation reaction, the molar ratio of the compound represented by the formula III-a to the compound represented by the formula II-a can be the conventional molar ratio of this type of reaction in the field, and the present invention is particularly Preferably it is 1:1-3:1, More preferably, it is 1.1:1-1.5:1 (for example, 1.3:1).

In the condensation reaction, the base can be a conventional base for this type of reaction in the field, and the present invention is particularly preferably an organic base, more preferably N,N-diisopropylethylamine.

In the condensation reaction, the molar ratio of the base to the compound shown in formula II-a can be the conventional molar ratio of this type of reaction in the field, and the present invention is particularly preferably 10:1 to 30: 1, more preferably 15:1 to 25:1 (for example, 21:1).

In the condensation reaction, the condensing agent may be a conventional condensing agent for this type of reaction in the field, and in the present invention, it is particularly preferably 1-n-propylphosphoric anhydride.

In the condensation reaction, the molar ratio of the condensing agent to the compound represented by formula II-a can be the conventional molar ratio of this type of reaction in the field, and is particularly preferably 1:1 to 3 in the present invention. : 1, more preferably 1.2:1 to 2:1 (for example, 1.5:1).

In the condensation reaction, the reaction temperature of the condensation reaction can be the conventional reaction temperature of this type of reaction in the art, and is particularly preferably room temperature in the present invention.

In the condensation reaction, the reaction time of the condensation reaction can be the conventional reaction time of this type of reaction in the field, and in the present invention, it is particularly preferably 8-20 hours, and more preferably 10-14 hours (for example, 12 hours).

In the condensation reaction, after the condensation reaction is completed, a post-processing step may be further included. The post-processing steps can be concentrating the reaction solution, extracting by phase separation, concentrating and purifying the organic phase. The reagents used in the phase separation extraction can be dichloromethane and 10% sodium carbonate aqueous solution. The purification method can be silica gel column purification. The eluent used in the silica gel column purification can be dichloromethane/methanol/triethylamine (volume ratio can be 20:1:0.5).

In an embodiment of the present invention, the method for preparing the compound represented by I-a is the method for preparing the compound represented by formula I. The preparation method of the compound shown in the formula I comprises the following steps: in a solvent, the compound shown in the formula II and the compound shown in the formula III are subjected to the following condensation under the action of a base and a condensing agent. reaction to obtain the compound shown in the formula I,

wherein, R ¹ , R ² , X, n and the carbon atoms marked with * are as described above.

In the condensation reaction, the solvent may be a conventional solvent for this type of reaction in the art, and in the present invention, it is particularly preferably a nitrile solvent (eg, acetonitrile).

In the condensation reaction, the molar concentration of the compound represented by the formula II in the solvent can be the conventional molar concentration of this type of reaction in the field, and is particularly preferably 0.01-0.5 mol/L in the present invention, and further It is preferably 0.05 to 0.15 mol/L (for example, 0.09 mol/L).

In the described condensation reaction, the mol ratio of the compound shown in the formula III and the compound shown in the formula II can be the conventional mol ratio of this type of reaction in this area, and the present invention is particularly preferably 1: 1 to 3:1, more preferably 1.1:1 to 1.5:1 (for example, 1.3:1).

In the condensation reaction, the molar ratio of the base to the compound shown in formula II can be the conventional molar ratio of this type of reaction in the field, and the present invention is particularly preferably 10:1 to 30:1, More preferably, it is 15:1 to 25:1 (for example, 21:1).

In the condensation reaction, the molar ratio of the condensing agent to the compound represented by formula II can be the conventional molar ratio of this type of reaction in the field, and the present invention is particularly preferably 1:1 to 3:1 , and more preferably 1.2:1 to 2:1 (for example, 1.5:1).

The present invention also provides a pharmaceutical composition, which includes the compound shown in formula Ia, its cis-trans isomer, its pharmaceutically acceptable salt, its solvate, its pharmaceutically acceptable Solvates of salts, tautomers thereof, or polymorphs thereof, and pharmaceutically acceptable excipients.

In one embodiment of the present invention, the pharmaceutical composition includes the compound shown in formula I, its cis-trans isomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient .

The present invention also provides a compound represented by formula Ia, its cis-trans isomer, its pharmaceutically acceptable salt, its solvate, its pharmaceutically acceptable salt solvate, Use of its tautomer, its polymorphic form, or said pharmaceutical composition in the preparation of CDK kinase inhibitor.

In one embodiment of the present invention, the CDK kinase inhibitor is a CDK7 kinase inhibitor.

In one embodiment of the present invention, the compound shown in formula I, its cis-trans isomer or a pharmaceutically acceptable salt thereof, or the application of the pharmaceutical composition in the preparation of a CDK kinase inhibitor .

The present invention also provides a compound shown in Ia, its cis-trans isomer, its pharmaceutically acceptable salt, its solvate, the solvate of its pharmaceutically acceptable salt, its mutual Use of the variant, or its polymorphic form, or the described pharmaceutical composition in the preparation of a medicament for preventing and/or treating CDK-related diseases.

In one embodiment of the present invention, the compound shown in formula I, its cis-trans isomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition is used in the preparation of prevention and/or treatment and CDK The use of medicines for related diseases.

In one embodiment of the present invention, the CDK-related disease is a CDK7-related disease, preferably a tumor, such as lung adenocarcinoma, lung cancer or breast cancer.

The present invention also provides a compound shown in formula II-a,

R ¹ , R ² , X, Y, R ⁴ , Z, L, ⁿ , na and carbon atoms marked with * are as previously described.

In one embodiment of the present invention, the compound represented by the formula II-a is the compound represented by the formula II,

In one embodiment of the present invention, the compound represented by formula II-a is

The present invention also provides a compound of formula IV-a1 or IV-a2,

R ¹ , R ² , X, Y, R ⁴ , Z, ⁿ , na and carbon atoms marked with * are as previously described.

In one embodiment of the present invention, the compound of formula IV-a1 is the compound of formula IV,

In one embodiment of the present invention, the compound represented by formula IV-a1 or IV-a2 is

In the present invention, room temperature refers to 10 to 30°C.

On the basis of conforming to common knowledge in the art, the above preferred conditions can be combined arbitrarily to obtain preferred examples of the present invention.

A pharmaceutically acceptable salt can be an acid addition salt with a pharmaceutically acceptable acid. Examples of pharmaceutically acceptable salt acids include inorganic acids such as nitric acid, boric acid, hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid; and organic acids such as oxalic acid, maleic acid, succinic acid, and citric acid . Non-limiting examples of salts of the compounds of the present invention include, but are not limited to, hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, 2-hydroxyethanesulfonate, phosphate, hydrogen phosphate , acetate, adipate, alginate, aspartate, benzoate, bisulfate, butyrate, camphorate, camphorsulfonate, digluconate, glycerophosphate Salt, Hemisulfate, Heptanoate, Caproate, Formate, Succinate, Fumarate, Maleate, Ascorbate, Isethionate, Salicylic Acid salt, mesylate, mesitylene sulfonate, naphthalene sulfonate, nicotinate, 2-naphthalene sulfonate, oxalate, pamoate, pectate, persulfate, 3 - Phenylpropionate, picrate, pivalate, propionate, trichloroacetate, trifluoroacetate, phosphate, glutamate, bicarbonate, p-toluenesulfonate, Undecanoate, lactate, citrate, tartrate, gluconate, mesylate, ethanedisulfonate, benzenesulfonate, and p-toluenesulfonate.

The choice of pharmaceutically acceptable adjuvants varies with the route of administration and the characteristics of action, and is usually a filler, diluent, binder, wetting agent, disintegrating agent, lubricant, emulsifying agent or suspending agent.

"Solvate" refers to a substance formed by crystallizing a compound with a solvent (including, but not limited to, water, methanol, ethanol, etc.). Solvates are divided into stoichiometric and non-stoichiometric solvates.

"Pharmaceutically acceptable salt solvate" refers to a compound with a pharmaceutically acceptable (relatively non-toxic, safe, suitable for patient use) acid or base, solvent (including but not limited to: water, methanol, ethanol, etc.) ) combined to form a substance, wherein the pharmaceutically acceptable salt has the same meaning as the term "pharmaceutically acceptable salt" above and the solvent is either stoichiometric or non-stoichiometric. Solvates of pharmaceutically acceptable salts include, but are not limited to, hydrochloride monohydrate.

"Tautomer" means that different functional group isomers are in dynamic equilibrium at room temperature and can be converted into each other quickly. A chemical equilibrium of tautomers can be achieved if tautomers are possible (eg, in solution). For example, proton tautomers (also referred to as proton tautomers) include interconversions by migration of protons, such as keto-enol isomerization and imine-enamine isomerization. A specific example of keto-enol tautomerization is the interconversion between two tautomers, pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

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

The term "alkyl" refers to a straight or branched chain alkyl group having the specified number of carbon atoms (eg, _C1 - _C6 ). Alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl Wait.

The term "cycloalkyl" refers to a saturated monocyclic cyclic group consisting of only carbon atoms having the specified number of carbon atoms (eg, C3 _- _C6 ). Cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

The term "heterocycloalkyl" refers to a specified number of ring atoms (eg, 5 to 8 members), a specified number of heteroatoms (eg, 1, 2, or 3), a specified heteroatom species (N, O, and S) one or more of), which are monocyclic, bridged, or spirocyclic, and each ring is saturated. Heterocycloalkyl groups include, but are not limited to, azetidinyl, tetrahydropyrrolyl, tetrahydrofuranyl, morpholinyl, piperidinyl, and the like.

in the structural fragment

It means that the structural fragment is connected to other fragments in the molecule through this site. E.g,

means cyclohexyl.

The term "plurality" refers to 2, 3 or 3.

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

The positive improvement effect of the present invention is that the compound of the present invention has better CDK7 inhibitory activity, cell activity and tumor growth inhibitory activity.

detailed description

The present invention is further described below by way of examples, but the present invention is not limited to the scope of the described examples. The experimental methods that do not specify specific conditions in the following examples are selected according to conventional methods and conditions, or according to the product description.

In the following preparations and examples, abbreviations are explained:

LDA: lithium diisopropylamide; rt: room temperature; DMSO: dimethyl sulfoxide; HOAc: acetic acid; EtOH: ethanol; DCM: dichloromethane; MTBE: methyl tert-butyl ether; ACN: acetonitrile; DIPEA: N,N-diisopropylethylamine; mCPBA: m-chloroperoxybenzoic acid; T3P: ₁ -n-propylphosphoric anhydride; Chloroform: chloroform; MgMeBr: methylmagnesium bromide; Boc: tert-butyloxy carbonyl.

Room temperature means 10-30 degreeC.

Preparation Example 1

Isovaleronitrile (compound 1, 25 g, 330 mmol) was slowly added dropwise to a solution of LDA in tetrahydrofuran (1.3 M, 300 ml) at -78°C. After the dropwise addition, the solution was continued to stir for 20 minutes, and then slowly added a solution of ethyl formate (compound 2, 30ml, 377mmol) in tetrahydrofuran (100ml) at -78°C, and the total dropwise addition time was controlled at 40 minutes. The reaction solution was at -78°C Stirring was continued for 45 minutes, then slowly warmed to room temperature and stirring was continued for 18 hours. The reaction was quenched with water (50 ml), adjusted to pH about 3 with aqueous hydrochloric acid (4N), then extracted with ethyl acetate (100 ml), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was washed with n-heptane Alkane was slurried, the suspension was filtered, and the filter cake was dried to give the product 2-formyl-3-methylbutyronitrile (compound 3, 23 g, 84%). ¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.72(s, 1H), 3.26(m, 1H), 2.55(m, 1H), 1.20(d, J=6.9Hz, 3H), 1.04(d, J=6.9Hz, 3H).

2-Formyl-3-methylbutyronitrile (compound 3, 9.97g, 90mmol), hydrazine hydrate (5.68ml, 117mmol) and acetic acid (9.02ml, 158mmol) were dissolved in ethanol (250ml), the whole solution was heated at 85°C and react for 20 hours. It was then cooled to room temperature and concentrated. The concentrate was diluted with saturated sodium carbonate solution to pH around 8 and extracted with dichloromethane (3 x 100 ml). The organic phase was washed with saturated brine, then dried over anhydrous magnesium sulfate, filtered and concentrated to dryness to obtain compound 4 as a product. ¹ H NMR (400MHz, CDCl ₃ ) δ 7.12 (d, J=0.7Hz, 1H), 2.69 (pd, J=6.9, 0.7Hz, 1H), 1.21 (s, 3H), 1.19 (s, 3H) .

Compound A1 (5.8 g, 1.1 eq) was slowly added dropwise to a dichloromethane solution of compound 4 (5 g, 1 eq) at 0°C. After the dropwise addition was completed, the reaction solution was slowly warmed to room temperature and stirring was continued for 10 hours. The reaction became a suspension, then concentrated, the residue was slurried with MTBE (30 ml), then filtered and dried to give the product compound A2 (4 g, 40%) LCMS: [M+H] ⁺ 257.0.

Compound A2 was dissolved in acetonitrile ACN (100ml), then potassium carbonate (5.4g, 2.5eq) was added, the reaction system was warmed to 70°C and stirring was continued for 4 hours. After cooling to room temperature, the reaction solution was concentrated, and then the pH was adjusted to about 3 with aqueous hydrochloric acid (1N). The suspension was filtered, the solid was washed with water, and then dried to give the product compound A3 (3 g, 92%); LCMS: [M+H] ⁺ 211.0.

To a suspension of compound A3 (1g, 1eq) in ethanol (20ml), sodium hydroxide aqueous solution (2M, 5ml, 2eq) was added at 0°C, stirred for 15 minutes, and then MeI (0.85g, 1.3eq) was added. The entire suspension was stirred at room temperature for a further 4 hours, then concentrated to remove most of the solvent, the residue was diluted with water and aqueous hydrochloric acid HCl (2N, 10ml) was added at 0°C. The suspension was filtered, and the solid was dried to obtain the product compound A4 (1 g, 94%). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ7.92 (s, 1H), 3.02 (p, J=6.9 Hz, 1H), 2.55(s, 3H), 1.27(s, 3H), 1.26(s, 3H). LCMS: [M+H] ⁺ 225.0.

A suspension of compound A4 (1 g, 4.46 mmol) in POCl ₃ (6 ml, 15 eq) was added to a sealed tube, DIPEA (1.5 ml, 2 eq) was added at 0°C, and the reaction solution was heated to 100°C. After stirring for 4 hours, the reaction was cooled to room temperature and then concentrated, the residue was diluted with ice water, the suspension was then filtered, the solid was washed with water and then dried to give the product compound A5. LCMS: [M+H] ⁺ 242.97.

To a mixture of A5 (3.05g, 12.5mmol, 1eq) and 1-Boc-4-aminopiperidine (2.5g, 12.5mmol, 1eq) in acetonitrile (80ml) was added DIPEA (4.4ml, 2eq) at 0°C , the reaction was warmed to room temperature and continued to stir for 2 hours, concentrated to remove acetonitrile, the residue was diluted with water, and the suspension was filtered and dried to obtain product D1. LCMS: [M+H] ⁺ 407.2.

The product of the previous step was dissolved in dichloromethane (100 ml), then mCPBA (5.4 g, 33 mmol, 2.6 eq) was added, and the whole suspension was stirred at room temperature for 12 hours. Then filtered, the filtrate was washed with aqueous sodium hydroxide solution (2N), the organic phase was further washed with water, then washed with saturated brine, finally dried with anhydrous sodium sulfate, filtered and concentrated to obtain product D3 (5 g, 91%).

¹ H NMR (400MHz, chloroform-d) δ7.95(s, 1H), 6.72(d, J=8.4Hz, 1H), 4.52-4.34(m, 1H), 4.23-4.02(m, 2H), 3.36 (s, 3H), 3.24 (hept, J=7.1Hz, 1H), 2.99 (t, J=12.5Hz, 2H), 2.21–2.07 (m, 2H), 1.58 (qd, J=12.2, 11.7, 4.6 Hz, 2H), 1.48 (s, 9H), 1.35 (d, J=6.9Hz, 6H). LCMS: [M+H] ⁺ 439.1.

To a solution of D3 (4g, 9.13mmol) in tetrahydrofuran (100ml) was slowly added MgMeBr (1M, 27ml, 3eq) at room temperature, then continued to stir for 2 hours, quenched with water (3eq) after the reaction was completed. The mixture was concentrated, the residue was extracted with dichloromethane/water phase, the organic phase was dried with sodium carbonate, then concentrated, and separated and purified by silica gel column, the eluent was ethyl acetate/n-heptane (2:1) to obtain the product D4 (3.4 g, 99%).

¹ H NMR (400MHz, chloroform-d) δ 7.80(s, 1H), 6.31(d, J=8.3Hz, 1H), 4.38-4.22(m, 1H), 4.22-4.03(m, 2H), 3.22 (hept, J=6.9Hz, 1H), 2.98(t, J=12.5Hz, 2H), 2.53(s, 3H), 2.09(dd, J=12.7, 3.7Hz, 2H), 1.55(td, J= 12.0, 4.3Hz, 2H), 1.48 (s, 9H), 1.32 (d, J=6.9Hz, 6H). LCMS: [M+H] ⁺ 375.2.

To a solution of D4 (3.4 g, 9 mmol) in dichloromethane (50 ml) was added a solution of hydrochloric acid in isopropanol (5 M, 6 ml), and stirring was continued at room temperature for 2 hours. Concentrated to dryness, the residue was extracted with dichloromethane/10% aqueous sodium carbonate solution, and the organic phase was concentrated to obtain product D5 (2.4 g, 99%).

¹ H NMR (400MHz, chloroform-d)δ7.80(s,1H),6.35(d,J=8.4Hz,1H),4.32-4.16(m,1H),3.29-3.11(m,3H),2.90 – 2.78 (m, 2H), 2.53 (s, 3H), 2.17–2.11 (m, 2H), 1.60 (tt, J=11.2, 5.7Hz, 2H), 1.32 (d, J=6.9Hz, 6H). LCMS: [M+H] ⁺ 275.2.

In a toluene solution (10ml) of tricarbonyl chloride (0.16g, 0.4eq), pyridine (0.07ml, 1eq) was added at 0°C, stirring was continued for 10 minutes, and then compound 5 (0.25g, 1.34mmol, 1eq) was added The solution of dichloromethane (3 ml) was slowly warmed to room temperature, and stirring was continued for 12 hours. The obtained suspension was filtered, and the filtrate containing compound 6 was retained and the next reaction was carried out.

The above solution containing compound 6 was added to a solution of D5 (0.3g, 0.8eq) and triethylamine (0.3ml) in dichloromethane (20ml) at room temperature, stirred for 1 hour, the reaction solution was concentrated, and the residue was washed with silica gel Column separation and purification [eluent dichloromethane/ethyl acetate (2:1)] gave compound D6 (0.45 g, 86%).

¹ H NMR (400MHz, chloroform-d)δ7.80(s,1H),6.32(d,J=8.2Hz,1H),5.25(s,1H),4.36-3.97(m,3H),3.60-3.39 (m, 4H), 3.22 (hept, J=7.0Hz, 1H), 3.04 (s, 2H), 2.53 (s, 3H), 2.19–2.02 (m, 4H), 1.46 (s, 11H), 1.32 ( d, J=6.9 Hz, 6H). LCMS: [M+H] ⁺ 488.3.

Preparation Example 2

Referring to the method for preparing D6 in Preparation Example 1, compound D8 was synthesized.

¹ H NMR (400MHz, chloroform-d) δ 7.81 (s, 1H), 6.36 (d, J=8.2 Hz, 1H), 5.11 (tt, J=6.7, 4.2 Hz, 1H), 4.24 (ddd, J =10.1,6.7,1.1Hz,2H),4.19–4.11(m,2H),3.92(dd,J=10.2,4.2Hz,2H),3.23(h,J=6.9Hz,1H),3.17–2.96( m, 2H), 2.53(s, 3H), 2.14(dd, J=13.1, 3.7Hz, 2H), 1.57(qd, J=11.7, 4.4Hz, 2H), 1.45(s, 9H), 1.32(d , J=6.9Hz, 6H). LCMS: [M+H] ⁺ 474.3.

Preparation Example 3

Referring to the method for preparing D6 in Preparation Example 1, compound D9 was synthesized.

¹ H NMR (400MHz, chloroform-d) δ 7.80 (s, 1H), 6.31 (d, J=8.2 Hz, 1H), 5.25 (s, 1H), 4.36–4.02 (m, 3H), 3.58–3.41 (m, 4H), 3.22 (hept, J=6.9Hz, 1H), 3.02 (d, J=13.6Hz, 2H), 2.53 (s, 3H), 2.09 (d, J=17.0Hz, 4H), 1.47 (s, 11H), 1.32 (d, J=6.9Hz, 6H).

Preparation Example 4

F1 (2.5g, 20mmol) and F2 (3.9g, 1.5eq) were in acetic acid (10ml), the reaction solution was stirred at 90 ^o C for 20 hours, then cooled to room temperature, and the resulting suspension was filtered, filtered with tert-methylbutyl Ethyl ether rinse and drying gave product F3 (3.4 g, 89%).

¹ H NMR (400MHz, DMSO-d ₆ ) δ 7.77(s, 1H), 5.52(s, 1H), 3.07(h, J=6.9Hz, 1H), 2.31(s, 3H), 1.22(d, J=6.9Hz, 6H). LCMS: [M+H] ⁺ 192.1.

Raw material F3 (3 g, 15.7 mmol) was dissolved in phosphine trichloride (15 ml, 10 eq), DIPEA (5.5 ml, 2 eq) was added at 0 °C, then stirred at 100 °C for 4 hours, cooled to room temperature, concentrated, and the residue was used Elution with ice water, the suspension was filtered, and dried to obtain the product F4. LCMS: [M+H] ⁺ 210.0.

To a solution of compound F4 (3.1, 14.8 mmol, 1 eq) and ammonia compound (3.3 g, 1.1 eq) in acetonitrile (80 ml), DIPEA (4.4 ml, 2 eq) was added at 0°C. The reaction was then stirred at room temperature for 2 hours, concentrated, diluted with water, and the suspension filtered and dried to give product F5 (5 g, 90%).

¹ H NMR (400MHz, chloroform-d) δ7.82(s, 1H), 6.12(d, J=8.2Hz, 1H), 5.78(s, 1H), 4.10(d, J=12.7Hz, 2H), 3.64(dddd,J=14.4,10.4,8.1,4.0Hz,1H),3.31(hept,J=6.9Hz,1H),3.00(t,J=12.3Hz,2H),2.52(s,3H),2.18 -2.00(m, 2H), 1.58(dtd, J=13.1, 11.0, 4.3Hz, 2H), 1.48(s, 9H), 1.34(d, J=6.9Hz, 6H). LCMS: [M+H] ⁺ 374.2.

To a solution of compound F5 (4 g, 10.7 mmol) in dichloromethane (50 ml) was added isopropanol hydrochloride (5 M, 10 ml), and stirring was continued at room temperature for 2 hours. Concentrated to dryness, the residue was extracted with dichloromethane/sodium carbonate (10% aqueous solution), and the organic phase was concentrated to obtain the product F6 (2.8 g, 98%).

¹ H NMR (400MHz, chloroform-d) δ 7.82 (s, 1H), 6.13 (d, J=8.2Hz, 1H), 3.59 (dddd, J=14.5, 10.4, 8.2, 4.1Hz, 1H), 3.30 (hept, J=6.9Hz, 1H), 3.19(dt, J=12.8, 3.9Hz, 2H), 2.78(ddd, J=13.0, 11.1, 2.6Hz, 2H), 2.51(s, 3H), 2.12( dt, J=13.3, 3.6 Hz, 2H), 1.56 (dtd, J=12.8, 10.8, 3.9 Hz, 2H), 1.34 (d, J=6.9 Hz, 6H). LCMS: [M+H] ⁺ 274.2.

Referring to the method of Preparation Example 1, compound F9 was synthesized.

¹ H NMR (400MHz, chloroform-d) δ 7.82 (s, 1H), 6.14 (d, J=8.2 Hz, 1H), 5.79 (s, 1H), 5.14–5.08 (m, 1H), 4.28–4.22 (m, 2H), 4.17–4.11 (m, 2H), 3.92 (dd, J=10.3, 4.2Hz, 2H), 3.75–3.61 (m, 1H), 3.31 (hept, J=7.0Hz, 1H), 3.09(s, 2H), 2.53(s, 3H), 2.22–2.08(m, 2H), 1.60(ddd, J=14.7, 10.7, 4.0Hz, 2H), 1.45(s, 9H), 1.34(d, J=6.9Hz, 6H). LCMS: [M+H] ⁺ 473.3.

Example 1

To the solution of D6 (0.13g, 0.27mmol) in dichloromethane (3ml) was added isopropanol hydrochloride (5M, 0.5ml), the reaction was stirred at room temperature for 2 hours, and concentrated to obtain crude product D6-1 which was directly used in the next reaction. D6-1: LCMS: [M+H]+388.3.

The above crude product was dissolved in acetonitrile (3ml)/DIPEA (0.5ml), then compound 7 (50mg, 1.3eq), T ₃ P (0.15g, 1.5eq) were added, DIPEA (0.5ml) was added, and the reaction solution was continued at room temperature Stirred for 12 hours, concentrated, the residue was phase-separatedly extracted with dichloromethane/sodium carbonate (10% aqueous solution), the organic phase was concentrated, separated on a silica gel column, and the eluent was dichloromethane/methanol/triethylamine (20:1:0.5) The product D7 was obtained (80 mg, 60%).

¹ H NMR (400MHz, DMSO-d ₆ )δ8.20(s,1H), 6.79-6.63(m,2H), 5.23-5.14(m,1H), 4.42-4.33(m,1H), 3.95-3.81 (m, 4H), 3.72–3.56 (m, 2H), 3.54–3.35 (m, 1H), 3.28 (p, J=6.7Hz, 1H), 3.09–2.84 (m, 4H), 2.72 (dd, J =4.9,1.6Hz,6H),2.54(s,3H),2.25–2.01(m,2H),1.83(d,J=12.3Hz,2H),1.67(dddd,J=18.7,11.7,7.4,4.2 Hz, 2H), 1.26 (d, J=6.8 Hz, 6H). LCMS: [M+H] ⁺ 499.3.

Example 2

Referring to the method for preparing D7 in Example 1, compound D10 was synthesized.

D9-1:LCMS:[M+H] ⁺ 388.2.

D10: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.18 (s, 1H), 6.79–6.62 (m, 2H), 5.19 (ddt, J=22.2, 5.2, 2.8 Hz, 1H), 4.38–4.34 (m, 1H), 3.88–3.76 (m, 4H), 3.71–3.35 (m, 4H), 3.24 (h, J=6.8Hz, 1H), 3.09–2.82 (m, 3H), 2.72 (dd, J =5.0,1.7Hz,6H),2.52(s,3H),2.24–2.00(m,2H),1.83(d,J=12.3Hz,2H),1.68(tq,J=12.3,7.5,5.7Hz, 2H), 1.26 (d, J=6.9Hz, 6H). LCMS: [M+H] ⁺ 499.3.

Example 3

Referring to the method for preparing D7 in Example 1, compound D11 was synthesized.

D8-1:LCMS:[M+H] ⁺ 374.2.

D11: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.16 (s, 1H), 6.66 (dt, J=15.4, 7.0 Hz, 1H), 6.43 (d, J=15.4 Hz, 1H), 5.11 ( tt, J=6.8, 4.1Hz, 1H), 4.56 (dd, J=9.9, 6.9Hz, 1H), 4.39–4.33 (m, 1H), 4.26 (dd, J=11.4, 6.8Hz, 1H), 4.20 –4.16(m,1H),4.05(d,J=7.1Hz,3H),3.89–3.81(m,3H),3.23(p,J=6.9Hz,1H),3.10–2.83(m,2H), 2.71(d,J＝4.9Hz,6H),2.50(s,3H),1.85(t,J＝8.0Hz,2H),1.70(d,J＝12.6Hz,2H),1.26(d,J＝6.9 Hz, 6H). LCMS: [M+H] ⁺ 485.3.

Example 4

Referring to the method for preparing D7 in Example 1, using F9 as a raw material, compound F10 was synthesized.

F9-1:LCMS:[M+H] ⁺ 373.2.

F10: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.31 (s, 1H), 6.75–6.59 (m, 2H), 6.53–6.34 (m, 1H), 5.12 (tt, J=6.8, 4.1 Hz ,1H),4.68–4.49(m,1H),4.32–4.08(m,4H),3.91–3.77(m,6H),3.46–3.39(m,1H),3.03(qd,J=7.5,5.6Hz ,1H),2.71(d,J＝4.8Hz,6H),2.62(s,3H),1.91–1.82(m,2H),1.77(s,2H),1.27(d,J＝6.9Hz,6H) . LCMS: [M+H] ⁺ 484.3.

Example 5

Pyridine (0.18ml, 1eq) was added to a solution of tricarbonyl chloride (0.26g, 0.4eq) in toluene (10ml) at 0°C, and after stirring for 10 minutes, diethylamino-3-hydroxypiperidine (0.44g, 1eq) was slowly added Chloromethane (3ml) solution was gradually returned to room temperature and stirred for 2 hours. After filtration, the filtrate was retained for the next step.

The above solution was slowly added to a solution of the corresponding free amine (0.5g, 0.8eq) and triethylamine (0.76ml) in dichloromethane (20ml) at room temperature. The reaction was carried out at room temperature for 1 hour, concentrated, and separated on a silica gel column. The eluent was dichloromethane-ethyl acetate (2:1) to obtain the solid product 17c-3 (0.9 g, 95%).

To a solution of the above 17c-3 (0.9 g) in ethyl acetate (5 ml) was added hydrochloric acid-ethyl acetate solution (2M, 4 ml), followed by stirring at room temperature for 3 hours. The suspension was filtered, and the filter cake was dried to obtain a white product 17c-4. LCMS: [M+H] ⁺ 402.2; C ₂₀ H ₃₁ N ₇ O ₂

¹ H NMR (400MHz, DMSO-d ₆ )δ8.48(s,1H),7.98(s,1H),4.61(tt,J=8.5,4.0Hz,1H),4.27(s,1H),4.03( dt, J=13.8, 3.6Hz, 2H), 3.20 (q, J=7.1Hz, 1H), 3.13–2.80 (m, 6H), 2.39 (s, 3H), 1.87–1.58 (m, 7H), 1.41 (dtd,J=12.6,9.1,3.8Hz,2H),1.27(d,J=7.0Hz,6H).

Refer to synthetic compounds 17-71;

LCMS: [M+H] ⁺ 513.3; C ₂₆ H ₄₀ N ₈ O ₃

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.47 (d, J=8.5 Hz, 1H), 7.98 (s, 1H), 6.60 (d, J=2.7 Hz, 2H), 4.81 (dd, J= 7.7,3.9Hz,1H),4.27(s,1H),4.05(d,J=13.3Hz,2H),3.69(d,J=10.0Hz,2H),3.22(q,J=7.0Hz,1H) ,3.12–2.85(m,5H),2.39(s,3H),2.16(s,7H),1.84(d,J=12.4Hz,4H),1.73–1.44(m,4H),1.27(d,J =7.

Example 6 Synthesis of intermediate CP2-11

Synthesized with reference to compound A2. CP-2:LCMS:[M ⁺ H]+255.0561.

Synthesized with reference to synthetic compound A3. CP-3:LCMS:[M ⁺ H]+209.0506.

Synthesized with reference to synthetic compound A4. CP-4:LCMS:[M ⁺ H]+223.0443.

Synthesized with reference to compound A5. CP-5:LCMS:[M ⁺ H]+241.0057.

Synthesized with reference to synthetic compound D1. CP-6:LCMS:[M ⁺ H]+405.1374.

Synthesized with reference to compound D3. CP-7:LCMS:[M ⁺ H]+437.1976.

Synthesized with reference to compound D4. CP-8:LCMS:[M ⁺ H]+373.1788.

Synthesized with reference to compound D5. CP-9:LCMS:[M ⁺ H]+273.1736.

Synthesized with reference to compound D8. CP-10: C23H33N7O4; LCMS: [M+H] ⁺ 472.2164.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.54 (d, J=8.6 Hz, 1H), 7.87 (s, 1H), 5.02 (tt, J=6.7, 4.0 Hz, 1H), 4.27 (ttd, J=11.2, 8.4, 7.9, 4.2Hz, 1H), 4.14(s, 2H), 4.08–3.94(m, 3H), 3.58(s, 1H), 3.12–2.78(m, 2H), 2.39(s, 3H),1.85(tdd,J=11.5,7.2,4.3Hz,3H),1.66(s,2H),1.38(s,9H),0.92–0.81(m,2H),0.81–0.69(m,2H) .

Synthesized with reference to compound D8-1. CP-11:LCMS:[M ⁺ H]+372.2324.

Example 7

Synthesized with reference to compound D11. 17-86: C24H34N8O3; LCMS: [M+H] ⁺ 483.3055.

¹ H NMR (400MHz, DMSO-d ₆ )δ8.54(d,J=8.5Hz,1H),7.87(s,1H),6.67-6.53(m,1H),6.20-6.01(m,1H), 5.09 (tt, J=6.9, 4.0Hz, 1H), 4.60–4.45 (m, 1H), 4.34–4.17 (m, 2H), 4.17–3.94 (m, 3H), 3.83 (dd, J=11.4, 4.0 Hz, 1H), 3.09–2.94 (m, 4H), 2.39 (s, 3H), 2.14 (d, J=1.6Hz, 6H), 1.85 (ddd, J=14.6, 8.9, 3.7Hz, 3H), 1.66 (s, 2H), 0.92–0.82 (m, 2H), 0.79–0.69 (m, 2H).

Example 8 Synthesis of intermediates tBP-1 to 11

Synthesized with reference to compound 4. tBP-1:LCMS:[M+H] ⁺ 140.1108

Synthesized with reference to compound A2. tBP-2:LCMS:[M+H] ⁺ 271.1536.

Synthesized with reference to synthetic compound A3. tBP-3:LCMS:[M+H] ⁺ 225.0321.

Synthesized with reference to synthetic compound A4. tBP-4:LCMS:[M+H] ⁺ 239.0962.

Synthesized with reference to compound A5. tBP-5:LCMS:[M+H] ⁺ 257.061.

Synthesized with reference to synthetic compound D1. tBP-6:LCMS:[M+H] ⁺ 421.218.

Synthesized with reference to compound D3. tBP-7: C20H32N6O4S; LCMS: [M+H] ⁺ 453.2274.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.44 (d, J=8.4 Hz, 1H), 8.23 (s, 1H), 4.29 (dtt, J=11.7, 7.7, 4.3 Hz, 1H), 4.13– 3.87(m, 2H), 3.37(d, J=5.9Hz, 5H), 1.84(dd, J=12.9, 3.8Hz, 2H), 1.67(qd, J=12.6, 4.4Hz, 2H), 1.42(d ,J=3.0Hz,18H).

Synthesized with reference to compound D4. tBP-8:LCMS:[M+H] ⁺ 389.2596.

Synthesized with reference to compound D5. tBP-9:LCMS:[M+H] ⁺ 289.2111.

Synthesize compound D8 with reference to

tBP-10:LCMS:[M+H] ⁺ 488.2685

Synthesized with reference to compound D8-1. tBP-11: C19H29N7O2; LCMS: [M+H] ⁺ 388.2033.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.91(s, 1H), 8.66(d, J=15.9Hz, 1H), 8.46(d, J=8.6Hz, 1H), 7.86(s, 1H) ,5.12–4.94(m,1H),4.28–4.11(m,3H),3.96–3.86(m,3H),2.90(dt,J=49.9,12.9Hz,2H),2.32(s,3H),1.83 –1.71(m, 2H), 1.58(d, J=12.2Hz, 2H), 1.29(s, 9H).

Example 9

Synthesized with reference to compound D11. 17-87: C25H38N8O3; LCMS: [M+H] ⁺ 499.2964.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.59 (d, J=8.7 Hz, 1H), 7.99 (s, 1H), 6.71-6.58 (m, 1H), 6.16 (dt, J=15.4, 1.7 Hz, 1H), 5.14 (tt, J=6.9, 4.0Hz, 1H), 4.56 (d, J=8.3Hz, 1H), 4.39–4.24 (m, 2H), 4.22–4.03 (m, 3H), 3.89 (dd, J=11.3, 4.0Hz, 1H), 3.14–3.01(m, 4H), 2.46(s, 3H), 2.20(s, 6H), 1.89(d, J=12.4Hz, 2H), 1.72( s,2H),1.44(s,9H).

Example 10

Synthesized with reference to compound D7.

¹ H NMR (400MHz, chloroform-d) δ 7.80 (s, 1H), 6.35 (d, J=8.3 Hz, 1H), 5.96 (dt, J=35.5, 9.0 Hz, 1H), 5.32 (q, J = 5.1, 3.7Hz, 1H), 4.38–4.01 (m, 3H), 3.93–3.59 (m, 4H), 3.24–3.04 (m, 4H), 2.53 (s, 3H), 2.29 (s, 3H), 2.23–2.02 (m, 6H), 1.93–1.78 (m, 5H), 1.67–1.51 (m, 3H), 1.32 (d, J=6.9Hz, 6H).

LCMS: [M+H] ⁺ 543.3;

Example 11

Synthesized with reference to compound D7.

¹ H NMR (400MHz, chloroform-d)δ7.80(s,1H),6.46-6.28(m,1H),5.95(dd,J=35.5,9.2Hz,1H),5.38-5.26(m,1H) ,4.39–3.98(m,3H),3.93–3.60(m,4H),3.27–3.03(m,5H),2.53(s,3H),2.29(s,3H),2.23–2.00(m,6H) ,1.96–1.78(m,6H),1.70–1.53(m,3H),1.32(d,J=6.9Hz,6H).

LCMS: [M+H] ⁺ 543.3;

Example 12

Refer to the synthesis of compound D7.

¹ H NMR (400 MHz, chloroform-d) δ 7.81 (s, 1H), 6.33 (d, J=8.2 Hz, 1H), 6.03 (dd, J=35.6, 9.3 Hz, 1H), 5.21 (tt, J =6.7,4.2Hz,1H),4.71(d,J=5.4Hz,1H),4.48-4.26(m,3H),4.25-4.04(m,3H),3.29-2.96(m,5H),2.53( s, 3H), 2.30 (s, 3H), 2.15 (dd, J=12.8, 3.6Hz, 2H), 2.06–1.80 (m, 5H), 1.70–1.53 (m, 3H), 1.32 (d, J= 7.0Hz, 6H).

LCMS: [M+H] ⁺ 529.3.

Example 13 Synthesis of Intermediates 9-11

A solution of bromide 5 (5 g, 85%) in acetonitrile (4 ml) was added potassium carbonate (5.8 g, 1.5 eq) followed by morpholine (2.7 ml, 1.1 eq). Stir at 30 °C for 2 h. Cool to At room temperature, filtered and concentrated, purified by silica gel column, and the eluent (4:1 n-heptane-ethyl acetate) gave liquid product 8 (4.4 g, 83%).

¹ H NMR (400 MHz, chloroform-d) δ 6.94 (dt, J=15.7, 6.1 Hz, 1H), 6.01 (dt, J=15.8, 1.6 Hz, 1H), 3.74 (s, 3H), 3.74-3.69 (m, 4H), 3.13 (dd, J=6.1, 1.7Hz, 2H), 2.46 (dd, J=5.6, 3.7Hz, 4H).

Sodium hydroxide (2.4g, 2.5eq) was added to a solution of the above methyl ester product (4.4g, 23.7mmol) in tetrahydrofuran (10ml)/water (5ml), and the mixture was stirred at room temperature for 4h. After the reaction solution was concentrated, residual The liquid was extracted with methyl tert-butyl, leaving the aqueous phase and adjusting the pH to around 2. It was then concentrated to dryness and the residue was extracted with methanol, filtered and concentrated to dryness to give 9 as a white product.

A solution of bromide 5 (5 g, 85%) in acetonitrile (4 ml) was added potassium carbonate (5.8 g, 1.5 eq) followed by 1-methylpiperazine (3.4 ml, 1.1 eq). Stir at 30°C 2h. Cooled to room temperature, filtered and concentrated, purified by silica gel column, and the eluent (4:1 n-heptane-ethyl acetate) gave solid product 10 (3 g, 58%).

¹ H NMR (400 MHz, chloroform-d) δ 6.96 (dt, J=15.7, 6.2 Hz, 1H), 6.00 (dt, J=15.7, 1.6 Hz, 1H), 3.74 (s, 3H), 3.14 (dd , J=6.2, 1.7Hz, 2H), 2.54(d, J=70.6Hz, 8H), 2.29(s, 3H).

Sodium hydroxide (1.6g, 2.5eq) was added to a solution of the above methyl ester product (3g, 16.2mmol) in tetrahydrofuran (10ml)/water (5ml), and the mixture was stirred at room temperature for 4h. After the reaction solution was concentrated, the residual liquid Extract with methyl tert-butyl, keep the aqueous phase and adjust the pH to around 2. It was then concentrated to dryness and the residue was extracted with methanol, filtered and concentrated to dryness to give 11 as a white product.

Example 14

A solution of free amine compound D8-1 (100mg, 0.269mmol) in acetonitrile (4ml) was added diisopropylethylamine (0.5ml), then 2-fluoroacrylic acid (37mg, 1.5eq), propylphosphoric anhydride ( 0.26g, 1.5eq). The reaction solution was stirred at room temperature for 20 hours. After concentration, the residual liquid was extracted with dichloromethane-10% sodium carbonate solution to separate the phases. The organic phase was taken, concentrated and purified by silica gel column. Methane-ethyl acetate (2: ₁ ) to give white product _17-70 . LCMS: [ _M +H] ⁺ 446.2; _C21H28FN7O3

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.51 (d, J=8.6 Hz, 1H), 7.99 (s, 1H), 5.49 (dd, J=48.4, 3.6 Hz, 1H), 5.31 (dd, J=16.6, 3.6Hz, 1H), 5.11 (ddd, J=10.9, 6.8, 4.1Hz, 1H), 4.67 (s, 1H), 4.40–4.20 (m, 3H), 4.19–3.85 (m, 3H) ,3.16–2.81(m,3H),2.40(s,3H),1.85(d,J=11.3Hz,2H),1.68(s,2H),1.27(d,J=6.9Hz,6H).

Example 15

Refer to the synthesis of compounds 17-70:

LCMS: [M+H] ⁺ 527.3; C ₂₆ H ₃₈ N ₈ O ₄

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.48 (d, J=8.5Hz, 1H), 7.98 (s, 1H), 6.62-6.55 (m, 1H), 6.12 (dt, J=15.4, 1.6 Hz, 1H), 5.09 (tt, J=6.9, 4.0Hz, 1H), 4.52 (t, J=8.4Hz, 1H), 4.34–4.01 (m, 5H), 3.87–3.79 (m, 1H), 3.57 (q, J=4.1, 3.6Hz, 6H), 3.10–3.05(m, 3H), 2.40(s, 3H), 2.35(t, J=4.7Hz, 4H), 1.85(d, J=11.7Hz, 2H), 1.67(d, J=12.3Hz, 2H), 1.27(d, J=6.9Hz, 6H).

Example 16

Synthesized compounds 17-71 with reference to

_LCMS : [ _M +H] ⁺ _540.3 ; _C27H41N9O3

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.48 (d, J=8.6 Hz, 1H), 7.98 (s, 1H), 6.57 (dt, J=15.4, 6.2 Hz, 1H), 6.17-6.06 ( m, 1H), 5.08 (tq, J=6.6, 3.3, 2.5Hz, 1H), 4.48 (dt, J=24.4, 8.5Hz, 1H), 4.34–4.00 (m, 6H), 3.83 (d, J= 13.0Hz, 1H), 3.62(s, 8H), 3.07(ddd, J=6.9, 4.5, 3.0Hz, 3H), 2.39(s, 3H), 2.21–2.17(m, 4H), 1.85(d, J =12.6Hz,2H),1.67(d,J=12.1Hz,2H),1.27(d,J=6.9Hz,6H).

Example 17

To a solution of free amine D8-1 (100 mg, 0.269 mmol) in dichloromethane (3 ml)/diisopropylethylamine (0.15 ml) was added acryloyl chloride (25 mg, 1 eq). The reaction was stirred at room temperature for 30 min. After concentration Direct silica gel column separation and purification, eluent dichloromethane-ethyl acetate-triethylamine (1:1:0.05) to obtain white solid 17-73.

LCMS: [M+H]+428.2; C21H29N7O3

1H NMR (400MHz, DMSO-d6) δ8.48 (d, J=8.6Hz, 1H), 7.98 (s, 1H), 6.31 (dd, J=17.0, 10.3Hz, 1H), 6.11 (dd, J= 17.0, 2.2Hz, 1H), 5.69 (dd, J=10.3, 2.2Hz, 1H), 5.10 (tt, J=6.9, 4.0Hz, 1H), 4.54 (t, J=8.4Hz, 1H), 4.33– 4.13 (m, 3H), 4.08–4.01 (m, 2H), 3.95–3.77 (m, 1H), 3.15–2.83 (m, 3H), 2.40 (s, 3H), 1.92–1.79 (m, 2H), 1.67(d,J＝12.3Hz,2H),1.27(d,J＝6.9Hz,6H).

Example 18

Reference to synthetic compounds 17-73

LCMS: [M+H]+456.2; C23H33N7O3

1H NMR(400MHz, DMSO-d6)δ8.47(d,J=8.6Hz,1H),7.98(s,1H),6.89-6.68(m,1H),6.09(dd,J=16.7,2.4Hz, 1H), 5.67 (dd, J=10.5, 2.4Hz, 1H), 4.82 (tt, J=7.5, 3.8Hz, 1H), 4.39–4.19 (m, 1H), 4.05 (d, J=13.2Hz, 2H) ), 3.71(ddd, J=13.5, 7.6, 3.9Hz, 2H), 3.49(s, 2H), 3.07(p, J=6.9Hz, 1H), 3.01–2.86(m, 2H), 2.39(s, 3H), 1.84 (d, J=12.4Hz, 4H), 1.73–1.49 (m, 4H), 1.27 (d, J=6.9Hz, 6H).

Example 19

Referring to the synthesis of compound 17c-3, compound 17c-7 was obtained (0.16g, 75%). LCMS: [M+H]+488.4.

Referring to the synthesis of compound 17c-3, compound 17c-8 was obtained. LCMS: [M+H]+388.2.

Refer to synthetic compounds 17-71;

LCMS: [M+H]+499.3; C25H38N8O3

1H NMR(400MHz, DMSO-d6)δ8.48(t,J=8.2Hz,1H),7.98(s,1H),6.69-6.50(m,1H),6.19-5.98(m,1H),4.25( dt, J=31.5, 10.0Hz, 3H), 4.08–3.84 (m, 3H), 3.16–2.88 (m, 5H), 2.40 (d, J=2.9Hz, 3H), 2.15 (d, J=4.7Hz) ,6H),1.81(t,J＝15.4Hz,2H),1.63(s,4H),1.27(d,J＝6.9Hz,6H),1.07–0.90(m,2H).

Example 20

To a solution of 1-BOC-3-amino-3-methylacridine (85mg, 1eq.) in dichloromethane (3ml)/diisopropylethylamine (0.4ml) was added carbonyldiimidazole (78mg, 1.05eq. ), the reaction was stirred at room temperature for 2 hours, then the piperidinamine derivative (0.13 g, 1 eq.) was added and stirred at room temperature for 2 hours. After the reaction, it was directly purified and separated by silica gel column, and the eluent was dichloromethane-ethyl acetate (1:1) to obtain white solid 17c-5 (0.2 g).

Trifluoroacetic acid (1 ml) was added to a solution of 17c-5 (0.2 g) in dichloromethane DCM (5 ml), and the reaction was stirred at room temperature for 3 hours. After concentration, the residue was extracted with dichloromethane-10% sodium carbonate solution to separate the phases, the organic phase was taken, the organic phase was dried with anhydrous sodium carbonate, filtered and concentrated to obtain the product 17c-6. LCMS:[M+H]+387.2

Refer to synthetic compounds 17-71;

LCMS: [M+H]+498.3; C25H39N9O2

1H NMR(400MHz, DMSO-d6)δ8.43(d,J=8.5Hz,1H),7.98(d,J=5.0Hz,1H),7.89(t,J=6.3Hz,1H),6.56(dt ,J=15.5,6.0Hz,1H),6.15(dt,J=15.4,1.6Hz,1H),4.19(d,J=8.8Hz,1H),4.15(d,J=8.1Hz,1H),3.98 (dd, J=19.3, 9.7Hz, 1H), 3.80 (t, J=9.8Hz, 3H), 3.26 (dd, J=13.3, 6.7Hz, 1H), 3.13–2.98 (m, 3H), 2.97– 2.83(m, 4H), 2.39(s, 3H), 2.09(s, 6H), 1.88–1.57(m, 5H), 1.27(d, J=6.9Hz, 6H).

Example 21

Referring to the synthesis of compound 17c-5, compound 17c-11 (0.2g). LCMS: [M+H]+473.2

Compound 17c-6 was synthesized with reference to obtain compound 17c-12 (0.12g). LCMS: [M+H]+373.2

Refer to synthetic compounds 17-71;

LCMS: [M+H]+484.3; C24H37N9O2

1H NMR(400MHz, DMSO-d6)δ8.49(dd,J=8.6,4.7Hz,1H),8.06(t,J=6.0Hz,1H),7.99(d,J=2.4Hz,1H),6.57 (ddt, J=15.4, 9.1, 6.2Hz, 1H), 6.15–6.02 (m, 1H), 4.25 (q, J=9.1, 7.3Hz, 2H), 3.99 (qd, J=6.4, 4.2Hz, 2H) ), 3.84(t, J＝14.4Hz, 2H), 3.30-3.20(m, 1H), 3.14-2.72(m, 6H), 2.39(s, 3H), 2.13(d, J＝9.1Hz, 6H) ,1.85–1.54(m,4H),1.27(d,J＝6.9Hz,6H).

Example 22

Referring to the synthesis of compound 17c-5, compound 17c-13 (0.2g). LCMS: [M+H]+488.3

Referring to the synthesis of compound 17c-6, compound 17c-14 (0.12g). LCMS: [M+H]+388.2

Reference to synthetic compounds 17-73

LCMS:[M+H]+442.2

C22H31N7O3

1H NMR(400MHz, DMSO-d6)δ8.52(d,J=8.6Hz,1H),8.39(d,J=7.9Hz,1H),7.99(s,1H),6.32-6.01(m,2H) ,5.60(dd,J=9.7,2.6Hz,1H),4.63(p,J=7.4Hz,1H),4.40-4.17(m,1H),4.08-3.93(m,3H),3.21-2.76(m ,3H),2.40(d,J＝1.1Hz,5H),1.98(dt,J＝11.9,8.2Hz,2H),1.84(d,J＝12.0Hz,2H),1.76–1.57(m,2H) ,1.27(d,J=6.9Hz,6H).

Example 23

Diisopropylethylamine (0.6ml, 3eq) was added to a solution of methylsulfone D3 (0.5g, 1.14mmol) and 4-aminotetrahydropyran (0.12g, 1.1eq) in isopropanol (3ml), and the mixture was Stir in a sealed tube at 90°C for 18 hours. The crude product was obtained after concentration, which was used directly in the next step. Ga1,LCMS:[M+H]+460.319

Reference synthesis compound D5: Ga2, LCMS: [M+H]+360.2298

Ga3: Refer to synthetic compound D8

Ga3,LCMS:[M+H]+559.2604

Ga4, refer to synthetic compound D8-1

Ga4, C22H34N8O3; LCMS: [M+H]+459.3708

1H NMR(400MHz, DMSO-d6)δ8.13(d,J=70.6Hz,1H),7.69(s,1H),6.96(d,J=45.4Hz,1H),5.05(p,J=6.3Hz) ,1H),4.25–3.98(m,3H),3.88(ddt,J=15.8,11.9,5.3Hz,3H),3.69(dd,J=9.4,6.8Hz,1H),3.51(t,J=7.8 Hz, 2H), 3.40 (m, J=2.0Hz, 3H), 3.07–2.67 (m, 4H), 1.84 (d, J=13.6Hz, 4H), 1.74–1.57 (m, 2H), 1.49 (tq , J=11.9, 5.5, 4.5Hz, 2H), 1.22(d, J=7.1Hz, 6H).

17-83: Reference to synthetic compounds 17-73

C25H36N8O4; LCMS: [M+H]+513.2823

1H NMR (400MHz, DMSO-d6) δ8.14 (d, J=68.8Hz, 1H), 7.69 (s, 1H), 7.06–6.82 (m, 1H), 6.32 (dd, J=17.0, 10.3Hz, 1H), 6.11(dd, J＝17.0, 2.2Hz, 1H), 5.69(dd, J＝10.3, 2.2Hz, 1H), 5.09(tt, J＝7.0, 4.0Hz, 1H), 4.54(t, J =8.4Hz,1H),4.32–3.97(m,5H),3.87(td,J=10.5,9.0,4.4Hz,4H),3.37(d,J=4.5Hz,2H),2.90(h,J= 6.7Hz, 3H), 1.97–1.74 (m, 4H), 1.65 (s, 2H), 1.57–1.42 (m, 2H), 1.22 (d, J=7.0Hz, 6H).

17-82: Refer to the synthesis of compound D11

C28H43N9O4; LCMS: [M+H]+570.3378

1H NMR(400MHz, DMSO-d6)δ8.20(d,J=68.5Hz,1H),7.75(s,1H),7.02(d,J=44.9Hz,1H),6.65(dt,J=15.3, 6.2Hz, 1H), 6.16 (dd, J=15.4, 1.8Hz, 1H), 5.14 (tt, J=6.9, 4.0Hz, 1H), 4.58 (t, J=8.5Hz, 1H), 4.35–4.03 ( m, 5H), 3.93 (tdt, J=15.4, 11.9, 5.3Hz, 4H), 3.43 (td, J=11.6, 2.1Hz, 4H), 3.14–3.03 (m, 3H), 2.20 (s, 6H) ,1.90(d,J＝12.9Hz,4H),1.71(s,2H),1.62–1.49(m,2H),1.29(d,J＝7.0Hz,6H).

Effect Example 1 CDK7 inhibitory activity

For the test method of CDK7 inhibitory activity, refer to WO2015058140.

Commercially available CDK7 (eurofins, Cat.No, 14-476M, Lot.No.WAE0003), CTD3peptide (GL Biochem, Cat.No.346885, Lot.P160205-SY346885), ATP (Sigma, Cat.No. A7699-1G, CAS No. 987-65-5), DMSO (Sigma, Cat. No. D2650, Lot. No. 474382), EDTA (Sigma, Cat. No. E5134, CAS No. 60-00-4) Other reagents were used to determine the inhibitory rate of compounds on CDK7 at 200nM and 10nM concentrations. The results are shown in Table 1.

Table 1

The control compound X is the compound of Example 1 of US20190144456

The control compound Y is the compound of Example 3 of US20190144456

Effect Example 2 CDK1, 2, 4, 6, 9 inhibitory activity

The test method of CDK1, 2, 4, 6, 9 inhibitory activity refers to US2019144456A1.

Commercially available CDK1 (Millipore, Cat. No 14-450M, Lot. No 25729U), CDK4 (Carna, Cat. No 04-105, Lot. No 14CBS-0306P), CDK6 (Carna, Cat. No 04- 107, Lot.No 15CBS-0744C), CDK9 (Millipore, Cat.No 14-685M, Lot.NoWAB0200), CDK2 (eurofins, Cat.No 14-448M, Lot.No D7NN039U-G), Peptide FAM-P18 ( GL Biochem, Cat.No.114204, Lot.No.P080319-XY114202), Peptide FAM-P8 (GL Biochem, Cat.No.112396, Lot.No.P080327-XY112396), CTD3 peptide (GL Biochem, Cat.No. .SY346885,Lot.No.P160205-SY346885),ATP(Sigma,Cat.No.A7699-1G,CAS No.987-65-5),DMSO(Sigma,Cat.No.D2650,Lot.No.474382) , EDTA (Sigma, Cat. No. E5134, CAS No. 60-00-4), 96-well plate (Corning, Cat. No. 3365, Lot. No. 22008026), 384-well plate (Corning, Cat. No.3573, Lot.No.12608008) and other reagents to determine the inhibitory rate of compounds on CDK1, 2, 4, 6, and 9 at 1000 nM and 100 nM concentrations. The results are shown in Table 2.

Table 2

The control compound X used in the test is the compound in Example 1 of US20190144456.

Effect Example 3 The influence of the test substance on the in vitro proliferation activity of each cell was detected by CCK8 method

Take logarithmic growth phase cells for experiments. The cells were digested, counted, made into a cell suspension of 1×10 ⁵ cells/ml, seeded in a 96-well plate (100 μL/well), and placed in a 37°C, 5% CO ₂ incubator for 24 hours; each well The test substance containing the corresponding concentration was added, and a negative control group and a blank group were set up at the same time, with 3 duplicate wells in each group; after the plate was incubated in an incubator for 72 hours, the cell morphology of each group was observed under a microscope, and 10 μL of CCK8 solution was added to each well. Incubate for 4 hours in a cell incubator, measure the absorbance at 450 nm, and calculate the proliferation inhibition rate. The results are shown in Table 3.

table 3

Effects of Example 4 Compounds on the growth of xenografted tumors in nude mice

The tumors at the vigorous growth stage were taken, and the tumor cells were inoculated subcutaneously in the right armpit of BALB/c nude mice under sterile conditions, and the inoculation amount of the cells was 5×10 ⁶ . The diameter of the transplanted tumor in nude mice was measured with a vernier caliper. When the tumor grew to about 100 mm ³ , the tumor-bearing nude mice with good growth status and good uniformity of tumor size were selected for group administration. After the administration, the nude mice were sacrificed by de-neck, and the tumor mass was surgically removed and weighed. The evaluation index of anti-tumor activity was tumor inhibition rate (%). The calculation formula was: tumor inhibition rate (%)=(tumor weight in model group - administration Tumor weight in drug group/tumor weight in model group*100%)

Table 4 Effects of compounds on the growth of human lung adenocarcinoma cell NCI-H1299 nude mice xenograft tumor (Mean±SD, n=8)

组别group	剂量dose	起始动物数number of starting animals	终末动物数number of end animals	瘤重(g)Tumor weight (g)	抑瘤率(％)Tumor inhibition rate (%)
模型组model group	10ml/kg生理盐水10ml/kg normal saline	88	88	1.220±0.0691.220±0.069	－-
对照化合物XControl Compound X	20mg/kg20mg/kg	88	88	0.634±0.062 ^ 0.634±0.062 ^	53.453.4
化合物F10Compound F10	20mg/kg20mg/kg	88	88	0.501±0.034 ^ 0.501±0.034 ^	68.468.4
化合物D11Compound D11	20mg/kg20mg/kg	88	88	0.420±0.037 ^ 0.420±0.037 ^	70.170.1

Compared with model group, ^* p<0.05, ^** p<0.01.

According to the above data, the three compounds have a certain inhibitory effect on the growth of human lung adenocarcinoma cell NCI-H1299 nude mouse xenograft tumor. The order of antitumor activity against NCI-H1299 xenograft tumor-bearing nude mice from strong to weak is: compound D11>compound F10>control compound X.

Table 5 Effects of compounds on the growth of human lung cancer cell A549 nude mice xenograft tumor (Mean±SD, n=8)

组别group	剂量dose	起始动物数number of starting animals	终末动物数number of end animals	瘤重(g)Tumor weight (g)	抑瘤率(％)Tumor inhibition rate (%)
模型组model group	10ml/kg生理盐水10ml/kg normal saline	88	88	1.197±0.0581.197±0.058	－-
对照化合物XControl Compound X	20mg/kg20mg/kg	88	88	0.652±0.071 ^ 0.652±0.071 ^	55.755.7
化合物F10Compound F10	20mg/kg20mg/kg	88	88	0.537±0.047 ^ 0.537±0.047 ^	64.964.9
化合物D11Compound D11	20mg/kg20mg/kg	88	88	0.488±0.069 ^ 0.488±0.069 ^	68.068.0

Compared with model group, ^* p<0.05, ^** p<0.01.

According to the above data, the three compounds have a certain inhibitory effect on the growth of human lung cancer cell A549 nude mouse xenograft tumor. The order of antitumor activity against human lung cancer cell A549 xenograft tumor-bearing nude mice is: compound D11>compound F10>control compound X.

Table 6 Effects of compounds on the growth of human breast cancer cells MDA-MB-468 nude mice xenograft tumor (Mean±SD, n=8)

组别group	剂量dose	起始动物数number of starting animals	终末动物数number of end animals	瘤重(g)Tumor weight (g)	抑瘤率(％)Tumor inhibition rate (%)
模型组model group	10ml/kg生理盐水10ml/kg normal saline	88	88	1.606±0.0781.606±0.078	－-
对照化合物XControl Compound X	20mg/kg20mg/kg	88	88	0.702±0.054 ^* 0.702±0.054 ^*	52.652.6
化合物F10Compound F10	20mg/kg20mg/kg	88	88	0.611±0.066 ^ 0.611±0.066 ^	66.366.3
化合物D11Compound D11	20mg/kg20mg/kg	88	88	0.509±0.059 ^ 0.509±0.059 ^	62.962.9

Compared with model group, ^* p<0.05, ^** p<0.01.

According to the above data, the three compounds have a certain inhibitory effect on the growth of human breast cancer cell MDA-MB-468 nude mouse xenograft tumor. The order of antitumor activity against human breast cancer cells MDA-MB-468 xenograft tumor-bearing nude mice is: compound F10>compound D11>control compound X.

Although the specific embodiments of the present invention have been described above, those skilled in the art should understand that these are only examples, and various changes may be made to these embodiments without departing from the principle and essence of the present invention. Revise. Accordingly, the scope of protection of the present invention is defined by the appended claims.

Claims

A compound as shown in formula Ia, its cis-trans isomer, its pharmaceutically acceptable salt, its solvate, its pharmaceutically acceptable salt solvate, its tautomer or its polymorph,

wherein R 1 and R 2 are independently C 1 -C 6 alkyl, one or more halogen-substituted C 1 -C 6 alkyl, C 3 -C 6 cycloalkyl or -NR 1-1 R 1- 2 ;

R 1-1 and R 1-2 are independently H or 5-8-membered heterocycloalkyl, and the heteroatom in the 5-8-membered heterocycloalkyl is one or more of N, S and O , the number is 1, 2 or 3;

X is N or CR 3 , R 3 is hydrogen or halogen;

Y is N or O;

R 4 is H or C 1 -C 6 alkyl;

n is 1 or 2;

n a is 1 or 2;

Z is N or CH;

L is NH or does not exist;

R 5 is H or halogen;

R 6 and R 7 are independently H, C 1 -C 6 alkyl, one or more R 6-1 substituted C 1 -C 6 alkyl, 5-8 membered heterocycloalkyl, or one or more R 6-2 -substituted 5-8-membered heterocycloalkyl, said 5-8-membered heterocycloalkyl, and, one or more R heteroatoms in the 6-2 -substituted 5-8-membered heterocycloalkyl is independently one or more of N, S and O, and the number is 1, 2 or 3;

R 6-1 is -NR 6-1-1 R 6-1-2 ;

R 6-1-1 and R 6-1-2 are independently H or C 1 -C 6 alkyl,

Alternatively, R 6-1-1 and R 6-1-2 together with the N to which they are attached form a 5-8 membered heterocycloalkyl or, one or more C 1 -C 6 alkyl substituted 5-8 membered heterocycle Alkyl; the heteroatoms in the 5-8-membered heterocycloalkyl group and the 5-8-membered heterocycloalkyl group substituted by one or more C 1 -C 6 alkyl groups are independently one of N, S and O One or more kinds, the number is 1, 2 or 3;

R 6-2 is C 1 -C 6 alkyl;

When n is 2, X is N;

The carbon atoms marked with * are chiral carbon in R configuration, chiral carbon in S configuration or achiral carbon.
The compound represented by formula Ia as claimed in claim 1, its cis-trans isomer, its pharmaceutically acceptable salt, its solvate, its pharmaceutically acceptable salt solvate, its interconversion Isomer or its polymorph, characterized in that when R 1 is C 1 -C 6 alkyl, the C 1 -C 6 alkyl is C 1 -C 4 alkyl, preferably ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, further preferably isopropyl or tert-butyl, for example isopropyl;

And/or, when R 1 is C 1 -C 6 alkyl substituted by one or more halogens, the halogen is fluorine, chlorine, bromine or iodine; the C 1 -C 6 alkyl is preferably C 1 -C 4 alkyl, further preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl;

And/or, when R 1 is a C 3 -C 6 cycloalkyl group, the C 3 -C 6 cycloalkyl group is a cyclopropanyl group, a cyclobutyl group, a cyclopentyl group or a cyclohexyl group, such as a cyclopropanyl group;

And/or, when R 2 is a C 1 -C 6 alkyl group, the C 1 -C 6 alkyl group is a C 1 -C 4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl base, n-butyl, isobutyl, sec-butyl or tert-butyl, more preferably methyl;

And/or, when R 2 is C 1 -C 6 alkyl substituted by one or more halogens, the halogen is fluorine, chlorine, bromine or iodine; the C 1 -C 6 alkyl is preferably C 1 -C 4 alkyl, further preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl;

And/or, when R 1-1 and R 1-2 are independently a 5-8 membered heterocycloalkyl, the 5-8 membered heterocycloalkyl is a 5-6 membered heterocycloalkyl, a heteroatom is O, and the number is 1; it is preferably a pyranyl group, such as

And/or, when R 3 is halogen, the halogen is fluorine, chlorine, bromine or iodine;

And/or, when R 4 is a C 1 -C 6 alkyl group, the C 1 -C 6 alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl radical, sec-butyl or tert-butyl, such as methyl;

and/or, when R 5 is halogen, the halogen is fluorine, chlorine, bromine or iodine, such as fluorine;

And/or, when R 6 and R 7 are independently C 1 -C 6 alkyl, or, one or more R 6-1 substituted C 1 -C 6 alkyl, said C 1 -C 6 alkyl In the alkyl group and the one or more R 6-1 substituted C 1 -C 6 alkyl groups, the C 1 -C 6 alkyl groups are independently methyl, ethyl, n-propyl, isopropyl, n-butyl radical, isobutyl, sec-butyl or tert-butyl, for example methyl;

And/or, when R 6-1-1 and R 6-1-2 are independently C 1 -C 6 alkyl, the C 1 -C 6 alkyl is independently methyl, ethyl, n-propyl radical, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example methyl;

And/or, when R 6-1-1 and R 6-1-2 together with the N to which they are attached form a 5-8 membered heterocycloalkyl, the 5-8 membered heterocycloalkyl is 5-6 membered Heterocycloalkyl, the heteroatom is N and/or O, and the number is 2; preferably morpholinyl, for example

And/or, when R 6-1-1 and R 6-1-2 together with the N to which they are attached form one or more C 1 -C 6 alkyl substituted 5-8 membered heterocycloalkanes, the 5 -8-membered heterocycloalkyl is a 5-6 membered heterocycloalkyl, the heteroatom is N, and the number is 1 or 2; preferably piperazinyl;

And/or, when R 6-1-1 and R 6-1-2 together with the N to which they are attached form one or more C 1 -C 6 alkyl substituted 5-8 membered heterocycloalkyl, the Ci- C6 - alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example methyl;

And/or, when said R 6 and R 7 are independently 5-8-membered heterocycloalkyl or 5-8-membered heterocycloalkyl substituted by one or more R 6-2 , said 5- 8-membered heterocycloalkyl and the 5-8-membered heterocycloalkyl in the one or more R 6-2 substituted 5-8-membered heterocycloalkyl are independently 5-6-membered heterocycloalkyl, The heteroatom is N and the number is 2, such as pyrrolidinyl;

And/or, when R 6-2 is C 1 -C 6 alkyl, the C 1 -C 6 alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl group, sec-butyl or tert-butyl, eg methyl.
The compound represented by formula Ia as claimed in claim 2, its cis-trans isomer, its pharmaceutically acceptable salt, its solvate, its pharmaceutically acceptable salt solvate, its interconversion Isomers or polymorphs thereof, characterized in that when R 2 is -NR 1-1 R 1-2 , one of R 1-1 and R 1-2 is H, and the other is a 5-8 membered heterocycle Alkyl; the -NR 1-1 R 1-2 is preferably

And/or, when R 6-1-1 and R 6-1-2 together with the N to which they are attached form one or more C 1 -C 6 alkyl substituted 5-8 membered heterocycloalkyl, the One or more C 1 -C 6 alkyl substituted 5-8 membered heterocycloalkyl is methyl substituted piperazinyl, for example

And/or, when said R 6 and R 7 are independently one or more R 6-2 substituted 5-8-membered heterocycloalkyl, said one or more R 6-2 substituted 5-membered heterocycloalkyl -8-membered heterocycloalkyl is methyl-substituted pyrrolidinyl, such as
The compound represented by formula Ia as claimed in claim 1, its cis-trans isomer, its pharmaceutically acceptable salt, its solvate, its pharmaceutically acceptable salt solvate, its interconversion Isomers or polymorphs thereof, characterized in that R 1 is C 1 -C 6 alkyl or C 3 -C 6 cycloalkyl, such as C 1 -C 6 alkyl;

and/or, R 2 is C 1 -C 6 alkyl or -NR 1-1 R 1-2 , such as C 1 -C 6 alkyl;

and/or, R 3 is hydrogen;

and/or, n a is 1;

and/or, Y is O;

and/or, Z is N and L is absent; or, Z is CH and L is NH;

And/or, R 6 and R 7 are independently H or, one or more R 6-1 substituted C 1 -C 6 alkyl; R 6-1 is -NR 6-1-1 R 6-1- 2 ; R 6-1-1 and R 6-1-2 are independently C 1 -C 6 alkyl, or, R 6-1-1 and R 6-1-2 together with the N to which they are attached form 5-8 membered heterocycloalkyl or, one or more C 1 -C 6 alkyl substituted 5-8 membered heterocycloalkyl;

and / or,
for
The compound represented by formula Ia as claimed in claim 1, its cis-trans isomer, its pharmaceutically acceptable salt, its solvate, its pharmaceutically acceptable salt solvate, its interconversion Isomers or polymorphs thereof, characterized in that it is the following scheme 1 or 2:

plan 1:

The compound shown in the formula I-a is the compound shown in the formula I-a1 or I-a2,

preferred;

In the compound shown in the formula I-a1:

R 1 and R 2 are independently C 1 -C 6 alkyl;

Y is O;

n is 2;

n a is 1;

Z is N;

L means does not exist;

R 6 and R 7 are independently one or more R 6-1 substituted C 1 -C 6 alkyl;

R 6-1 is -NR 6-1-1 R 6-1-2 ;

and, R 6-1-1 and R 6-1-2 are independently C 1 -C 6 alkyl;

In the compound shown in the formula I-a2:

R 1 is C 1 -C 6 alkyl;

R 2 is C 1 -C 6 alkyl or -NR 1-1 R 1-2 ;

X is N or CH;

Y is O;

Z is N;

L means does not exist;

R 5 is H;

R 6 and R 7 are independently H, C 1 -C 6 alkyl, or, one or more R 6-1 substituted C 1 -C 6 alkyl;

R 6-1 is -NR 6-1-1 R 6-1-2 ;

and, R 6-1-1 and R 6-1-2 are independently C 1 -C 6 alkyl;

Scenario 2:

The compound shown in the formula I-a is the compound shown in the formula I, its cis-trans isomer or a pharmaceutically acceptable salt thereof,

in,

R 1 and R 2 are independently C 1 -C 6 alkyl, or C 1 -C 6 alkyl substituted with one or more halogens;

X is N or CR 3 , R 3 is hydrogen or halogen;

n is 1 or 2;

The carbon atom marked with * is chiral carbon in R configuration, chiral carbon in S configuration or achiral carbon;

When n is 2, X is N;

Preferably, the compound represented by the formula I is the compound represented by the formula I-1 or the compound represented by the formula I-2,

In the compound shown in the formula I-1, R 1 and R 2 are independently C 1 -C 6 alkyl, or C 1 -C 6 alkyl substituted by one or more halogens; and, with * The labeled carbon atom is R-configuration chiral carbon, S-configuration chiral carbon or achiral carbon;

In the compound shown in the formula I-2, R 1 and R 2 are independently C 1 -C 6 alkyl, or C 1 -C 6 alkyl substituted by one or more halogens; and, X is N or CR 3 , and R 3 is hydrogen or halogen.
The compound represented by formula Ia as claimed in claim 1, its cis-trans isomer, its pharmaceutically acceptable salt, its solvate, its pharmaceutically acceptable salt solvate, its interconversion Isomer or its polymorph, characterized in that the compound shown in formula Ia is
The preparation method of the compound represented by formula Ia according to any one of claims 1-6, characterized in that, it comprises the following steps: in a solvent, mixing the compound represented by formula II-a with the compound represented by formula III The compound shown in -a is subjected to the condensation reaction shown below under the action of a base and a condensing agent to obtain the compound shown in the formula Ia,

Wherein, R 8 is halogen or hydroxyl; R 1 , R 2 , X, Y, R 4 , Z, L, n , R 5 , na , R 6 , R 7 and carbon atoms marked with * are as claimed in claim 1 Any of -6.
The preparation method of the compound shown in formula I-a as claimed in claim 7, is characterized in that,

Described solvent is nitrile solvent, such as acetonitrile;

And/or, the molar concentration of the compound represented by the formula II-a in the solvent is 0.01-0.5 mol/L, preferably 0.05-0.15 mol/L, for example 0.09 mol/L;

And/or, the molar ratio of the compound represented by the formula III-a to the compound represented by the formula II-a is 1:1 to 3:1, preferably 1.1:1 to 1.5:1 , such as 1.3:1;

And/or, the base is an organic base, preferably N,N-diisopropylethylamine;

And/or, the molar ratio of the base to the compound represented by formula II-a is 10:1-30:1, preferably 15:1-25:1, for example 21:1;

And/or, described condensing agent is 1-n-propyl phosphoric anhydride;

And/or, the molar ratio of the condensing agent to the compound represented by formula II-a is 1:1-3:1, preferably 1.2:1-2:1, for example 1.5:1;

And/or, the reaction temperature of described condensation reaction is room temperature;

And/or, the reaction time of the condensation reaction is 8 to 20 hours, preferably 10 to 14 hours, for example, 12 hours.
A pharmaceutical composition, it comprises the compound shown in formula Ia as described in any one of claim 1-6, its cis-trans isomer, its pharmaceutically acceptable salt, its solvate, its Solvates of pharmaceutically acceptable salts, tautomers or polymorphs thereof, and pharmaceutically acceptable excipients.
A compound shown in formula Ia as described in any one of claims 1-6, its cis-trans isomer, its pharmaceutically acceptable salt, its solvate, its pharmaceutically acceptable salt The solvate, its tautomer or its polymorphic form, or the application of the pharmaceutical composition according to claim 9 in the preparation of CDK kinase inhibitor; the CDK kinase inhibitor is preferably CDK7 kinase inhibitor agent.
A compound shown in formula Ia as described in any one of claims 1-6, its cis-trans isomer, its pharmaceutically acceptable salt, its solvate, its pharmaceutically acceptable salt The application of the solvate, its tautomer or its polymorphic form, or the pharmaceutical composition as claimed in claim 9 in the preparation of a medicine for preventing and/or treating CDK-related diseases; The relevant disease is preferably a CDK7-related disease, more preferably a tumor such as lung adenocarcinoma, lung cancer or breast cancer.
A compound of formula II-a, IV-a1 or IV-a2,

The definitions of R 1 , R 2 , X, Y, R 4 , Z, L, n , na and the carbon atoms marked with * are as described in any one of claims 1-6.
The compound represented by the formula II-a, IV-a1 or IV-a2 according to claim 12, wherein the compound represented by the formula II-a is the compound represented by the formula II; Formula IV-a1 is a compound shown in Formula IV;

wherein, R 1 , R 2 , X, n and the carbon atoms marked with * are as described in any one of claims 1-6.
The compound represented by formula II-a, IV-a1 or IV-a2 according to claim 12, wherein the compound represented by formula II-a is

The compound shown in the formula IV-a1 or IV-a2 is