CN116730979A

CN116730979A - Pol theta inhibitor

Info

Publication number: CN116730979A
Application number: CN202310230336.6A
Authority: CN
Inventors: 张学军; 李金平; 李群; 贾一民; 陈浩民; 陈登辉; 宋小波; 张辛; 杨俊�; 李莉娥
Original assignee: Humanwell Healthcare Group Co ltd; Wuhan Humanwell Innovative Drug Research and Development Center Ltd Co
Current assignee: Humanwell Healthcare Group Co ltd; Wuhan Humanwell Innovative Drug Research and Development Center Ltd Co
Priority date: 2022-03-11
Filing date: 2023-03-10
Publication date: 2023-09-12
Also published as: TW202341987A; WO2023169573A1

Abstract

The invention provides a Pol theta inhibitor. Specifically provides a heterocyclic compound shown in a formula I, a stereoisomer or pharmaceutically acceptable salt thereof, wherein m and n are integers from 0 to 4 respectively; ring K is a 5-6 membered heteroaromatic ring. The heterocyclic compound has good inhibition effect on Pol theta polymerase and can prevent or treat diseases or symptoms mediated by Pol theta.

Description

Pol theta inhibitor

Technical Field

The invention belongs to the field of medicines, and particularly relates to a Pol theta inhibitor.

Background

Synthetic mortality is an emerging direction of research in the field of antitumor, where treatment for DNA repair pathways is the focus of this patent. Part of tumor cells can cause deletion of a certain DNA repair pathway due to gene mutation, so that the tumor cells are excessively dependent on the existing DNA repair pathway, and the tumor cells can be specifically killed by targeted inhibition of the existing DNA repair pathway, but have no killing effect on normal somatic cells. In recent years, methods for treating mutant tumors of the breast ovarian cancer susceptibility gene (breast and ovarian cancer susceptibility gene, BRCA) by inhibitors of poly (ADP-ribose) polymerase have utilized a synthetic lethal mechanism targeting DNA repair defects.

DNA double strand breaks (DNA double strand breaks, DSBs) are one of the most severe DNA lesions. DNA double strand break repair (DNA double strand breaks repair, DSBR) in cells can be broadly divided into three pathways, one non-homologous end joining (NHEJ), one by homologous recombination (homologous recombination, HR) and the last by end joining (alt-EJ) mediated by DNA Polymerase θ (Polymerase θ, pol θ, POLQ) when NHEJ or HR is damaged, also known as microhomology-mediated end joining (MMEJ). Pol theta plays a central role in the process of micro-homology mediated end ligation. Pol theta has an N-terminal helicase domain and a C-terminal DNA polymerase domain. Studies have shown that the helicase domain of Pol theta promotes annealing of micro-homology, after which all outstanding bases are removed by nucleases and gaps are filled by Pol theta. Therefore, pol θ is receiving increasing attention as an important target for DNA repair defects. Pol theta is hardly expressed in normal tissues but is overexpressed in various tumor types (e.g., breast cancer, ovarian cancer, HNSCC, and lung cancer) and is associated with poor prognosis results. Studies have shown that when homologous recombination mediated repair is compromised (HRD), such as BRCA1 or BRCA2 mutations, pol theta is highly expressed and directs DSB repair toward Microhomology Mediated End Joining (MMEJ), opening the DNA repair process of MMEJ.

Since Pol theta is critical in homologous recombination repair defective (HRD) tumors, inhibition of Pol theta is a promising novel synthetic lethal therapeutic strategy.

Disclosure of Invention

It is an object of the present invention to provide a Pol theta inhibitor having a structure of formula I as shown, which is useful for inhibiting the activity of Pol theta polymerase, preventing or treating a disease or condition mediated by Pol theta.

The invention provides a heterocyclic compound shown in a formula I, a stereoisomer or pharmaceutically acceptable salt thereof:

wherein m is 0, 1, 2, 3 or 4;

n is 0, 1, 2, 3 or 4;

ring K is a 5-6 membered heteroaromatic ring;

R ₁ and R is ₂ Each independently is halogen, hydroxy, amino, cyano, unsubstituted or substituted with one or more R _a Substituted C ₁ -C ₆ Alkyl, unsubstituted or substituted by one or more R _a Substituted C ₁ -C ₆ Alkoxy, unsubstituted or substituted by one or more R _a Substituted 3-6 membered cycloalkyl;

R ₁ and R is ₂ Wherein each R is _a Each independently is deuterium, halogen, hydroxy, amino, cyano, C ₁ -C ₆ Alkyl or C ₁ -C ₆ A haloalkyl group;

R ₃ is hydrogen, unsubstituted or substituted by one or more R _a Substituted C ₁ -C ₆ An alkyl group;

R ₃ wherein each R is _a Each independently is deuterium,Halogen, hydroxy, amino, cyano, C ₁ -C ₆ Alkyl or C ₁ -C ₆ A haloalkyl group;

R ₄ 、R ₅ 、R ₆ And R is ₇ Each independently is hydrogen or R _b ，R ₈ Is R _b ；

Or, R ₄ 、R ₅ 、R ₆ 、R ₇ And R is ₈ Each independently is hydrogen or R _b And R is ₄ 、R ₅ 、R ₆ 、R ₇ And R is ₈ At least one of which is cyano;

or, R ₄ And R is ₅ Each independently is R _b ，

Or, R ₆ And R is ₇ Each independently is R _b ，

Or, R ₄ And R is ₅ Together with the C atom to which they are attached form a ring A, R ₆ 、R ₇ And R is ₈ Each independently is hydrogen or R _b ；

Or, R ₅ And R is ₆ Together with the C atom to which they are attached form a ring A, R ₄ 、R ₇ And R is ₈ Each independently is hydrogen or R _b ；

Or, R ₆ And R is ₇ Together with the C atom to which they are attached form a ring A, R ₄ 、R ₅ And R is ₈ Each independently is hydrogen or R _b ；

Or, R ₇ And R is ₈ Together with the C atom to which they are attached form a ring A, R ₄ 、R ₅ And R is ₆ Each independently is hydrogen or R _b ；

Each R is _b Each independently is halogen, hydroxy, amino, cyano, unsubstituted or R _a Substituted C ₁ -C ₆ Alkyl, unsubstituted or substituted by R _a Substituted C ₁ -C ₆ Alkoxy, unsubstituted or substituted by R _a Substituted 3-6 membered cycloalkyl, unsubstituted or substituted by R _a Substituted 4-6 membered heterocycloalkyl;

each R is _b Each of said R _a Each independently deuterium, halogenHydroxy, amino, cyano, C ₁ -C ₆ Alkyl or C ₁ -C ₆ A haloalkyl group;

each of said rings A being unsubstituted or substituted by one or more R _a Substituted 3-8 membered cycloalkyl, unsubstituted or substituted by R _a Substituted 4-8 membered heterocycloalkyl;

in each ring A, the R _a Each independently is deuterium, halogen, hydroxy, amino, cyano, C ₁ -C ₆ Alkyl or C ₁ -C ₆ A haloalkyl group.

In one embodiment, in the heterocyclic compound of formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, certain groups in the heterocyclic compound of formula I are defined as follows, and the remaining groups are defined as described in any other embodiment (hereinafter referred to as "in one embodiment"): the heterocyclic compound shown in the formula I, a stereoisomer or pharmaceutically acceptable salt thereof:

wherein m is 0, 1, 2, 3, 4;

n is 0, 1, 2, 3, 4;

ring K is a 5-6 membered heteroaromatic ring;

R ₁ 、R ₂ each independently selected from: halogen, hydroxy, amino, cyano, unsubstituted or substituted by one or more R _a Substituted C ₁ -C ₆ Alkyl, unsubstituted or substituted by one or more R _a Substituted C ₁ -C ₆ Alkoxy, unsubstituted or substituted by one or more R _a Substituted 3-6 membered cycloalkyl;

when m is 2, 3, 4, said R ₁ The same or different;

when n is 2, 3 or 4, R is ₂ The same or different;

R ₄ 、R ₅ 、R ₆ 、R ₇ each independently is hydrogen or R _b ，R ₈ Is R _b ；

Or R is ₄ 、R ₅ 、R ₆ 、R ₇ 、R ₈ Each independently is hydrogen or R _b And R is ₄ 、R ₅ 、R ₆ 、R ₇ 、R ₈ At least one of which is cyano;

or R is ₄ And R is ₅ R being identical or different _b Or R is ₄ And R is ₅ Together with the C atom to which they are attached form a ring A, R ₆ 、R ₇ 、R ₈ Each independently is hydrogen or R _b ；

Or R is ₅ And R is ₆ Together with the C atom to which they are attached form a ring A, R ₄ 、R ₇ 、R ₈ Each independently is hydrogen or R _b ；

Or R is ₆ And R is ₇ R being identical or different _b Or R is ₆ And R is ₇ Together with the C atom to which they are attached form a ring A, R ₄ 、R ₅ 、R ₈ Each independently is hydrogen or R _b ；

Or R is ₇ And R is ₈ Together with the C atom to which they are attached form a ring A, R ₄ 、R ₅ 、R ₆ Each independently is hydrogen or R _b ；

Wherein any one of the above R _b Each independently is halogen, hydroxy, amino, cyano, unsubstituted or R _a Substituted C ₁ -C ₆ Alkyl, unsubstituted or substituted by R _a Substituted C ₁ -C ₆ Alkoxy, unsubstituted or substituted by R _a Substituted 3-6 membered cycloalkyl, unsubstituted or substituted by R _a Substituted 4-6 membered heterocycloalkyl;

the ring A is 3-8 membered cycloalkyl which is unsubstituted or substituted by one or more Ra, 4-8 membered heterocycloalkyl which is unsubstituted or substituted by Ra;

any of the above Ra are each independently deuterium, halogen, hydroxy, amino, cyano, C ₁ -C ₆ Alkyl, C ₁ -C ₆ A haloalkyl group;

when Ra is plural, the Ra is the same or different.

In one embodiment, the heterocyclic compound shown in the formula I, the stereoisomer or the pharmaceutically acceptable salt thereof,

Wherein m is 0, 1, 2, 3, 4;

n is 0, 1, 2, 3, 4;

ring K is a 5-6 membered heteroaromatic ring;

when m is 2, 3, 4, said R ₁ The same or different;

when n is 2, 3 or 4, R is ₂ The same or different;

the ring A being unsubstituted or substituted by one or more R _a Substituted 3-8 membered cycloalkyl, unsubstituted or substituted by R _a Substituted 4-8 membered heterocycloalkyl;

any one of the above R _a Each independently is halogen, hydroxy, amino, cyano, C ₁ -C ₆ Alkyl, C ₁ -C ₆ A haloalkyl group;

when R is _a When there are a plurality of R _a The same or different.

In one embodiment, the ring K is a 5-6 membered N-containing heteroaryl ring; preferably, ring K is a pyridine ring, pyrimidine ring, pyrazine ring or pyridazine ring.

In one embodiment, the heterocyclic compound of formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, has structure II

Wherein m is 0, 1, 2, 3, 4;

n is 0, 1, 2, 3, 4;

when m is 2, 3, 4, said R ₁ The same or different;

when n is 2, 3 or 4, R is ₂ The same or different;

Or R is ₇ And R is ₈ To which they are attachedThe C atoms together form a ring A, R ₄ 、R ₅ 、R ₆ Each independently is hydrogen or R _b ；

Wherein any one of the above R _b Each independently is halogen, hydroxy, amino, cyano, unsubstituted or R _a Substituted C ₁ -C ₆ Alkyl, unsubstituted or substituted by R _a Substituted C ₁ -C ₆ An alkoxy group;

when R is _a When there are a plurality of R _a The same or different.

In one embodiment, any one of the plurality of R _a Substituted means 2, 3 or 4R _a Substitution; when R is _a When the number of the R is 2, 3 or 4, the R is _a The same or different.

In one embodiment, the heterocyclic compound of formula II, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

wherein m and n are integers of 0 to 4 respectively;

R ₁ 、R ₂ each independently selected from: halogen, hydroxy, amino, cyano, C ₁ -C ₆ Alkyl, 3-6 membered cycloalkyl;

the R is ₁ Or R is ₂ Optionally substituted with a substituent selected from the group consisting of: halogen, hydroxy, amino, C ₁ -C ₆ An alkyl group;

when R is ₁ When there are a plurality of R ₁ The same or different; when R is ₂ When there are a plurality of R ₂ The same or different;

R ₃ is hydrogen, C ₁ -C ₆ Alkyl, C ₁ -C ₆ Deuterated alkyl or C ₁ -C ₆ A haloalkyl group;

R ₄ 、R ₅ 、R ₆ 、R ₇ each independently is hydrogen or C ₁ -C ₆ Alkyl, R ₈ Is C ₁ -C ₆ An alkyl group;

or R is ₄ And R is ₅ Together with the C atom to which they are attached form a ring A, R ₆ 、R ₇ 、R ₈ Each independently is hydrogen or C ₁ -C ₆ An alkyl group;

or R is ₅ And R is ₆ Together with the C atom to which they are attached form a ring A, R ₄ 、R ₇ 、R ₈ Each independently is hydrogen or C ₁ -C ₆ An alkyl group;

or R is ₆ And R is ₇ Together with the C atom to which they are attached form a ring A, R ₄ 、R ₅ 、R ₈ Each independently is hydrogen or C ₁ -C ₆ An alkyl group;

or R is ₇ And R is ₈ Together with the C atom to which they are attached form a ring A, R ₄ 、R ₅ 、R ₆ Each independently is hydrogen or C ₁ -C ₆ An alkyl group;

the ring A is 3-8 membered cycloalkyl or 4-8 membered heterocycloalkyl;

The ring a is optionally substituted with a substituent selected from the group consisting of: halogen, hydroxy, amino, cyano, C ₁ -C ₆ Alkyl, C ₁ -C ₆ A haloalkyl group.

m and n are integers of 0 to 4 respectively;

the R is ₁ Or R is ₂ Optionally substituted with a substituent selected from the group consisting of: halogen, hydroxyRadicals, amino radicals, C ₁ -C ₆ An alkyl group;

R ₃ is hydrogen, C ₁ -C ₆ Alkyl or C ₁ -C ₆ A haloalkyl group;

the ring A is 3-8 membered cycloalkyl or 4-8 membered heterocycloalkyl;

In one embodiment, the heterocyclic compound of formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, has structure Ia or Ib

Wherein m, n, R ₁ 、R ₂ 、R ₃ 、R ₄ 、R ₅ 、R ₆ 、R ₇ And R is ₈ As defined in any one of the present inventions.

In one aspect of the present invention,having the structure->R ₁₁ Is C ₁ -C ₃ Alkyl, R ₁₂ Is C ₁ -C ₃ A haloalkyl group;

preferably, the method comprises the steps of,having the structure->

In one aspect of the present invention,having the structure->R ₂₁ 、R ₂₂ 、R ₂₃ Halogen which is the same or different; preferably, said R ₂₁ 、R ₂₂ 、R ₂₃ Each independently F or Cl;

preferably, the method comprises the steps of,having the structure->

In one embodiment, R ₄ And R is ₅ Together with the C atom to which they are attached form a ring A, R ₆ 、R ₇ 、R ₈ Each independently is hydrogen.

In one embodiment, R ₄ And R is ₅ Together with the C atom to which they are attached form a ring A, R ₆ 、R ₇ 、R ₈ Each independently is hydrogen, hydroxy, C ₁ -C ₆ Alkyl, C ₁ -C ₆ A haloalkyl group;

or R is ₅ And R is ₆ Together with the C atom to which they are attached form a ring A, R ₄ 、R ₇ 、R ₈ Each independently is hydrogen, hydroxy, C ₁ -C ₆ Alkyl, C ₁ -C ₆ A haloalkyl group;

or R is ₆ And R is ₇ Together with the C atom to which they are attached form a ring A, R ₄ 、R ₅ 、R ₈ Each independently is hydrogen, hydroxy, C ₁ -C ₆ Alkyl, C ₁ -C ₆ A haloalkyl group;

or R is ₇ And R is ₈ Together with the C atom to which they are attached form a ring A, R ₄ 、R ₅ 、R ₆ Each independently is hydrogen, hydroxy, C ₁ -C ₆ Alkyl, C ₁ -C ₆ A haloalkyl group.

In one embodiment, R ₄ 、R ₅ Together with the C atom to which they are attached form a ring A, R ₆ 、R ₇ 、R ₈ Each independently is hydrogen;

or R is ₅ And R is ₆ Together with the C atom to which they are attached form a ring A, R ₄ 、R ₇ 、R ₈ Is hydrogen;

or R is ₆ And R is ₇ Together with the C atom to which they are attached form a ring A, R ₄ 、R ₅ 、R ₈ Is hydrogen;

or R is ₇ And R is ₈ Together with the C atom to which they are attached form a ring A, R ₄ 、R ₅ 、R ₆ Is hydrogen;

the ring A is 3-6 membered cycloalkyl or 4-8 membered heterocycloalkyl;

preferably, the ring a is selected from: cyclopropane, cyclobutane, cyclopentane, cyclohexane;

preferably, the 4-8 membered heterocycloalkyl has 1, 2 or 3 heteroatoms selected from N, O or S; when the hetero atom is plural, the hetero atoms may be the same or different.

In one embodiment, R ₄ 、R ₅ 、R ₆ 、R ₇ Is hydrogen, R ₈ Is C ₁ -C ₃ An alkyl group.

In one aspect, the C ₁ -C ₆ Haloalkyl is C ₁ -C ₆ A fluoroalkyl group.

In one aspect, the C ₁ -C ₃ Haloalkyl is C ₁ -C ₃ A fluoroalkyl group.

In one embodiment, R ₄ And R is ₅ R being identical or different _b ，R ₆ 、R ₇ 、R ₈ Each independently is hydrogen, hydroxy, halogen, C ₁ -C ₆ Alkyl, C ₁ -C ₆ A haloalkyl group;

or R is ₆ And R is ₇ R being identical or different _b ，R ₄ 、R ₅ 、R ₈ Each independently is hydrogen, hydroxy, C ₁ -C ₆ Alkyl, C ₁ -C ₆ A haloalkyl group;

R _b is halogen, hydroxy, cyano, C ₁ -C ₆ An alkyl group;

preferably, R _b Fluorine, chlorine, hydroxyl, cyano, methyl, ethyl, propyl.

In one embodiment, the heterocyclic compound of formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is characterized by having structure Ic, id, or Ie

Wherein the ring K, m, n, R ₁ 、R ₂ 、R ₃ 、R ₄ 、R ₅ 、R ₆ 、R ₇ And R is ₈ As defined in any one of the present inventions.

In one embodiment, in Ic, R ₅ And R is ₆ Together with the C atom to which they are attached form ring A.

In one embodiment, in Id, R is ₅ And R is ₆ Together with the C atom to which they are attached form a ring a; or R is ₆ And R is ₇ Together with the C atom to which they are attached form a ring a; or R is ₇ And R is ₈ Together with the C atom to which they are attached form ring A.

In one embodiment, in Ie, R ₄ And R is ₅ Together with the C atom to which they are attached form a ring a; or R is ₅ And R is ₆ Together with the C atom to which they are attached form ring A.

In one embodiment, any of the above ring a is a 3-6 membered cycloalkyl or 4-8 membered heterocycloalkyl; preferably, the ring a is selected from: cyclopropane, cyclobutane, cyclopentane, cyclohexane.

In one embodiment, the 4-8 membered heterocycloalkyl has 1, 2 or 3 heteroatoms selected from N, O or S; when the hetero atom is plural, the hetero atoms may be the same or different.

In one embodiment, R ₃ Is methyl.

In one embodiment, R ₃ Is deuterated methyl (-CD) ₃ )。

In one embodiment, in Ic, id or Ie, R ₄ 、R ₅ 、R ₆ 、R ₇ And R is ₈ Each independently is hydroxy, methyl, fluoromethyl, cyano, cyclopropyl.

In one embodiment, ring K is a 5-or 6-membered N-containing heteroaryl ring; preferably, ring K is a pyridine ring, pyrimidine ring, pyrazine ring, pyridazine ring.

In one aspect of the present invention,having the structure->

In one embodiment, m is 2 or 3, such as 2.

In one embodiment, n is 1 or 3, e.g., 3.

In one embodiment, the ring K is a 5-6 membered heteroaromatic ring, wherein the heteroatom is N and the number of heteroatoms is 1 or 2, e.g., the ring K is pyridine.

In one embodiment, each R ₁ Each independently is halogen, unsubstituted or substituted with one or more R _a Substituted C ₁ -C ₆ Alkyl or 3-6 membered cycloalkyl, preferably each R ₁ Each independently is halogen, unsubstituted or substituted with one or more R _a Substituted C ₁ -C ₆ Alkyl, e.g. each R ₁ Each independently is halogen, trifluoromethyl, methyl or tridentate methyl, as well as, for example, each R ₁ Each independently is unsubstituted or substituted with one or more R _a Substituted C ₁ -C ₆ An alkyl group.

In one embodiment, when m is 2, each R ₁ Located ortho or para to the heteroatom in ring K.

In one embodiment, when m is 3, each R ₁ Adjacent and in the ortho, meta or para position to the heteroatom in ring K.

In one embodiment, each R ₁ Wherein each R is _a Each independently is halogen or deuterium, e.g., each R ₁ Wherein each R is _a Each independently is halogen.

In one embodiment, R ₂ Each independently is halogen.

In one embodiment, R ₃ Unsubstituted or substituted by one or more R _a Substituted C ₁ -C ₆ Alkyl radicals, e.g. R ₃ Is C ₁ -C ₆ Alkyl, e.g. R ₃ To be covered by one or more R _a Substituted C ₁ -C ₆ An alkyl group.

In one embodiment, R ₃ Wherein each R is _a Each independently deuterium.

In one embodiment, each R _b Each independently is halogen, hydroxy, cyano, 3-6 membered cycloalkyl or is unsubstituted or substituted with one or more R _a Substituted C ₁ -C ₆ Alkyl, preferably each R _b Each independently is halogen, hydroxy or C ₁ -C ₆ Alkyl, e.g. each R _b Each independently is hydroxy or C ₁ -C ₆ An alkyl group.

In one embodiment, each R _b Wherein each R is _a Each independently is halogen.

In one embodiment, each ring A is independently a 3-8 membered cycloalkyl.

In one embodiment, R ₄ 、R ₅ 、R ₆ And R is ₇ Each independently is hydrogen or R _b ，R ₈ Is C ₁ -C ₆ An alkyl group.

In one embodiment, R ₅ 、R ₆ 、R ₇ And R is ₈ Each independently is hydrogen or R _b And R is ₄ Is cyano.

In one embodiment, R ₄ 、R ₅ 、R ₇ And R is ₈ Each independently is hydrogen or R _b And R is ₆ Is cyano.

In one embodiment, R ₄ And R is ₅ Each independently is R _b ，R ₆ 、R ₇ And R is ₈ Each independently is R _b Or hydrogen, or a combination of hydrogen and,

for example, R ₄ And R is ₅ Each independently is R _b ，R ₆ And R is ₇ Each independently is hydrogen, C ₁ -C ₆ Alkyl or hydroxy, R ₈ Is a hydrogen gas which is used as a hydrogen gas,

preferably, R ₄ And R is ₅ Each independently is R _b ，R ₆ And R is ₇ Each independently is hydrogen or hydroxy, R ₈ Is hydrogen.

In one embodiment, R ₆ And R is ₇ Each independently is R _b ，R ₄ 、R ₅ And R is ₈ Each independently is R _b Or hydrogen, or a combination of hydrogen and,

for example, R ₆ And R is ₇ Each independently is R _b ，R ₄ And R is ₅ Each independently is hydrogen, C ₁ -C ₆ Alkyl or hydroxy, R ₈ Is hydrogen.

In one embodiment, R ₄ And R is ₅ Together with the C atom to which they are attached form a ring A, R ₆ And R is ₇ Each independently is hydrogen or hydroxy, R ₈ Is hydrogen, e.g. R ₄ And R is ₅ Together with the C atom to which they are attached form a ring A, R ₆ And R is ₇ Each independently is hydrogen, R ₈ Is hydrogen.

In one embodiment, R ₅ And R is ₆ Together with the C atom to which they are attached form a ring A, R ₄ And R is ₇ Each independently is hydrogen or hydroxy, R ₈ Is hydrogen.

In one embodiment, R ₆ And R is ₇ Together with the C atom to which they are attached form a ring A, R ₄ And R is ₅ Each independently is hydrogen or hydroxy, R ₈ Is hydrogen.

In one embodiment, in ring K, the 5-6 membered heteroaromatic ring is one or more heteroatoms selected from N, O and S, and the number of heteroatoms is 1, 2 or 3, for example, in ring K, the 5-6 membered heteroaromatic ring is a 6 membered heteroaromatic ring, preferably the 5-6 membered heteroaromatic ring is N, more preferably the number of heteroatoms in the 5-6 membered heteroaromatic ring is 1.

In one embodiment, R ₁ And R is ₂ Wherein each halogen is independently fluorine, chlorine, bromine or iodine, such as chlorine or fluorine.

In one embodiment, R ₁ And R is ₂ In said formula (I), said R being unsubstituted or substituted by one or more R _a Substituted C ₁ -C ₆ C in alkyl ₁ -C ₆ Alkyl is each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl.

In one embodiment, R ₁ And R is ₂ In said formula (I), said R being unsubstituted or substituted by one or more R _a Substituted C ₁ -C ₆ C in alkoxy ₁ -C ₆ Alkoxy groups are each independently methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy or tert-butoxy.

In one embodiment, R ₁ And R is ₂ Wherein each 3-6 membered cycloalkyl is independently cyclopropanyl, cyclobutyl, cyclopentyl or cyclohexenyl, e.g. cyclopropanyl.

In one embodiment, R ₁ And R is ₂ Wherein each R is _a Wherein each halogen is independently fluorine, chlorine, bromine or iodine, such as fluorine.

In one embodiment, R ₁ And R is ₂ Wherein each R is _a In the above, the C ₁ -C ₆ Alkyl is each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl.

In one embodiment, R ₁ And R is ₂ Wherein each R is _a In the above, the C ₁ -C ₆ Haloalkyl is each independently C, substituted with 1, 2 or 3 halogens (e.g., fluorine, chlorine, bromine or iodine) ₁ -C ₆ Alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl).

In one embodiment, R ₃ In said formula (I), said R being unsubstituted or substituted by one or more R _a Substituted C ₁ -C ₆ C in alkyl ₁ -C ₆ Alkyl is each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl.

In one embodiment, R ₃ Wherein each R is _a Wherein each halogen is independently fluorine, chlorine, bromine or iodine, such as fluorine.

In one embodiment, R ₃ Wherein each R is _a In the above, the C ₁ -C ₆ Alkyl is each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl.

In one embodiment, R ₃ Wherein each R is _a In the above, the C ₁ -C ₆ Haloalkyl groups are each independently 1, 2 or 3 halogens (e.g., fluorine, chlorine, bromine) Or iodine) substituted by C ₁ -C ₆ Alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl).

In one embodiment, R ₄ 、R ₅ 、R ₆ 、R ₇ And R is ₈ In the above, the C ₁ -C ₆ Alkyl is each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl.

In one embodiment, each R _b Wherein each halogen is independently fluorine, chlorine, bromine or iodine, such as fluorine.

In one embodiment, each R _b In said, unsubstituted or R _a Substituted C ₁ -C ₆ C in alkyl ₁ -C ₆ Alkyl is each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl.

In one embodiment, each R _b In said, unsubstituted or R _a Substituted C ₁ -C ₆ C in alkoxy ₁ -C ₆ Alkoxy groups are each independently methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy or tert-butoxy.

In one embodiment, each R _b In said, unsubstituted or R _a The 3-6 membered cycloalkyl groups in the substituted 3-6 membered cycloalkyl groups are each independently cyclopropane, cyclobutane, cyclopentane or cyclohexane groups, for example cyclopropane groups.

In one embodiment, each R _b In said, unsubstituted or R _a In the 4-6 membered heterocycloalkyl group in the substituted 4-6 membered heterocycloalkyl group, the hetero atom is one or more selected from N, O and S, and the hetero atom number is 1, 2 or 3.

In one embodiment, each R _b Wherein each R is _a Wherein each halogen is independently fluorine, chlorine, bromine or iodine, such as fluorine.

In one embodiment, each R _b Wherein each R is _a In the above, the C ₁ -C ₆ Alkyl groups are each independently methyl, ethyl, n-propyl, isopropyl,N-butyl, isobutyl or tert-butyl.

In one embodiment, each R _b Wherein each R is _a In the above, the C ₁ -C ₆ Haloalkyl is each independently C, substituted with 1, 2 or 3 halogens (e.g., fluorine, chlorine, bromine or iodine) ₁ -C ₆ Alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl).

In one embodiment, in each ring A, the ring is unsubstituted or substituted with one or more R _a The 3-8 membered cycloalkyl group in the substituted 3-8 membered cycloalkyl group is a 3-6 membered cycloalkyl group, for example, the 3-6 membered cycloalkyl groups are each independently a cyclopropanyl group, a cyclobutyl group, a cyclopentanyl group or a cyclohexenyl group, preferably a cyclopropanyl group.

In one embodiment, in each ring A, the ring is unsubstituted or substituted with R _a The 4-8 membered heterocycloalkyl group in the substituted 4-8 membered heterocycloalkyl group is a 4-6 membered heterocycloalkyl group, preferably, the 4-6 membered heterocycloalkyl group has one or more hetero atoms selected from N, O and S and 1, 2 or 3 hetero atoms.

In one embodiment, each R in each ring A _a Wherein each halogen is independently fluorine, chlorine, bromine or iodine.

In one embodiment, each R in each ring A _a In the above, the C ₁ -C ₆ Alkyl is each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl.

In one embodiment, each R in each ring A _a In the above, the C ₁ -C ₆ Haloalkyl is each independently C, substituted with 1, 2 or 3 halogens (e.g., fluorine, chlorine, bromine or iodine) ₁ -C ₆ Alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl).

In one embodiment, when m is 2, 3, 4, the R ₁ The same or different.

In one embodiment, when n is 2, 3, 4, the R ₂ The same or different.

In one embodiment, when Ra is plural, the Ra is the same or different.

In one embodiment, the term "or" is merely "or" and represents a relationship between elements.

In one embodiment, when the alkyl is unsubstituted, the alkyl is unsubstituted.

In one embodiment, when the alkyl group is not described as isotopically substituted, the alkyl group is one that does not contain carbon and/or hydrogen isotopes.

In one embodiment, when the cycloalkyl or carbocyclyl is not described as an unsaturated or partially saturated carbocycle, the cycloalkyl or carbocyclyl is a saturated cycloalkyl or carbocyclyl.

In one embodiment, when the heterocycloalkyl is not described as an unsaturated or partially saturated heterocycloalkyl, the heterocycloalkyl is a saturated heterocycloalkyl.

In one embodiment, when the heteroaryl group is not described as a heteroaryl group fused to a benzene ring, the heteroaryl group is an unfused heteroaryl group.

In one embodiment, the stereoisomers are enantiomers or diastereomers.

m is 0, 1, 2, 3 or 4;

n is 0, 1, 2, 3 or 4;

the ring K is a 5-6 membered heteroaromatic ring, wherein in the 5-6 membered heteroaromatic ring, hetero atoms are N, and the number of the hetero atoms is 1 or 2;

each R is ₁ Each independently is halogen, unsubstituted or substituted with one or more R _a Substituted C ₁ -C ₆ Alkyl or 3-6 membered cycloalkyl;

each R is ₁ Wherein each R is _a Each independently is halogen or deuterium;

R ₂ each independently is halogen;

R ₃ unsubstituted or substituted by one or more R _a Substituted C ₁ -C ₆ An alkyl group;

R ₃ wherein each R is _a Each independently isDeuterium;

R ₄ 、R ₅ 、R ₆ and R is ₇ Each independently is hydrogen or R _b ，R ₈ Is C ₁ -C ₆ An alkyl group;

or, R ₅ 、R ₆ 、R ₇ And R is ₈ Each independently is hydrogen or R _b And R is ₄ Is cyano;

or, R ₄ 、R ₅ 、R ₇ And R is ₈ Each independently is hydrogen or R _b And R is ₆ Is cyano;

or, R ₄ And R is ₅ Each independently is R _b ，R ₆ And R is ₇ Each independently is hydrogen, C ₁ -C ₆ Alkyl or hydroxy, R ₈ Is hydrogen;

or, R ₆ And R is ₇ Each independently is R _b ，R ₄ And R is ₅ Each independently is hydrogen, C ₁ -C ₆ Alkyl or hydroxy, R ₈ Is hydrogen;

or, R ₄ And R is ₅ Together with the C atom to which they are attached form a ring A, R ₆ And R is ₇ Each independently is hydrogen or hydroxy, R ₈ Is hydrogen;

or, R ₅ And R is ₆ Together with the C atom to which they are attached form a ring A, R ₄ And R is ₇ Each independently is hydrogen or hydroxy, R ₈ Is hydrogen;

or, R ₆ And R is ₇ Together with the C atom to which they are attached form a ring A, R ₄ And R is ₅ Each independently is hydrogen or hydroxy, R ₈ Is hydrogen;

each R is _b Each independently is halogen, hydroxy, cyano, 3-6 membered cycloalkyl or is unsubstituted or substituted with one or more R _a Substituted C ₁ -C ₆ An alkyl group;

each R is _b Wherein each R is _a Each independently is halogen;

each ring a is independently a 3-8 membered cycloalkyl.

m is 2 or 3;

n is 3;

ring K is pyridine;

each R is ₁ Each independently is halogen, unsubstituted or substituted with one or more R _a Substituted C ₁ -C ₆ An alkyl group;

when m is 2, each R ₁ Ortho-or para-to the heteroatom in ring K, each R when m is 3 ₁ Adjacent and in the ortho, meta or para position to the heteroatom in ring K;

each R is ₁ Wherein each R is _a Each independently is halogen or deuterium;

R ₂ each independently is halogen;

R ₃ wherein each R is _a Each independently deuterium;

R ₄ and R is ₅ Each independently is R _b ，R ₆ And R is ₇ Each independently is hydrogen or hydroxy, R ₈ Is hydrogen;

or, R ₄ And R is ₅ Together with the C atom to which they are attached form a ring A, R ₆ And R is ₇ Each independently is hydrogen, R ₈ Is hydrogen;

each R is _b Each independently is halogen, hydroxy or C ₁ -C ₆ An alkyl group;

each ring a is independently a 3-8 membered cycloalkyl.

m is 2 or 3;

n is 3;

ring K is pyridine;

each R is ₁ Each independently is halogen, trifluoromethyl, methyl or tridentate methyl;

R ₂ each independently is halogen;

R ₃ wherein each R is _a Each independently deuterium;

each R is _b Each independently is hydroxy or C ₁ -C ₆ An alkyl group;

each ring a is independently a 3-8 membered cycloalkyl.

m is 2;

n is 3;

ring K is pyridine;

each R is ₁ Each independently is unsubstituted or substituted with one or more R _a Substituted C ₁ -C ₆ An alkyl group;

when m is 2, each R ₁ Ortho or para to the heteroatom in ring K;

each R is ₁ Wherein each R is _a Each independently is halogen;

R ₂ each independently is halogen;

R ₃ is C ₁ -C ₆ An alkyl group;

each R is _b Each independently is hydroxy or C ₁ -C ₆ An alkyl group;

each ring a is independently a 3-8 membered cycloalkyl.

m is 2 or 3;

n is 3;

ring K is pyridine;

each R is ₁ Wherein each R is _a Each independently is halogen or deuterium;

R ₂ each independently is halogen;

R ₃ to be covered by one or more R _a Substituted C ₁ -C ₆ An alkyl group;

R ₃ wherein each R is _a Each independently deuterium;

each R is _b Each independently is hydroxy or C ₁ -C ₆ An alkyl group;

each ring a is independently a 3-8 membered cycloalkyl.

In one aspect of the present invention,is that

In one aspect of the present invention,is->

In one embodiment, ring K is pyridine or pyrimidine.

In one embodiment, R ₁ Is methyl, trifluoromethyl (-CF) ₃ ) Difluoromethyl (-CHF) ₂ ) Cyclopropane group, fluorine or tridentate methyl (-CD) ₃ )。

In one embodiment, R ₂ Is fluorine or chlorine.

In one embodiment, R ₃ Is methyl or tridentate methyl (-CD) ₃ )。

In one embodiment, R ₄ And R is ₅ Each independently is hydrogen, fluoro, hydroxy, methyl, cyclopropyl, trifluoromethyl, cyano or difluoromethyl.

In one embodiment, R ₄ And R is ₅ Together with the C atom to which they are attached form a ring a, which is cyclopropane.

In one embodiment, R ₅ And R is ₆ Together with the C atom to which they are attached form a ring a, which is cyclopropane.

In one embodiment, R ₆ And R is ₇ Each independently is hydrogen, hydroxy, methyl or cyano.

In one embodiment, R ₈ Is hydrogen or methyl.

In a certain scheme, the heterocyclic compound shown in the formula I is any one of the following compounds:

the invention provides a pharmaceutical composition comprising a heterocyclic compound shown in the formula I, a stereoisomer or a pharmaceutically acceptable salt thereof and optionally

i) One or more active agents; and/or

ii) a pharmaceutically acceptable carrier.

The invention provides a pharmaceutical composition, which comprises a heterocyclic compound shown in the formula I, a stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and/or diluent.

The invention provides an application of a heterocyclic compound shown in the formula I, a stereoisomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition, which comprises the following steps:

1) Inhibiting Pol theta activity;

2) Preventing and treating Pol theta mediated diseases;

3) Preparing a medicament, pharmaceutical composition or formulation for inhibiting Pol theta activity;

4) Preparing a medicament, a pharmaceutical composition or a preparation for preventing and treating a Pol theta mediated disease;

preferably, the Pol θ mediated disease is cancer.

The invention provides a heterocyclic compound shown in the formula I, a stereoisomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition for preventing or treating cancers.

The present invention also provides a method of treating a disease comprising administering to a patient a therapeutically effective amount of at least one of a heterocyclic compound of formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above.

In one embodiment, the disease is a disease mediated by Pol θ, e.g., a disease characterized by overexpression of Rolq.

In one embodiment, the disease is a cell proliferative disorder; preferably, the cell proliferative disorder is cancer.

In one embodiment, the patient is a mammal, preferably a human.

In one embodiment, examples of cancers (and benign counterparts thereof) that may be treated (or inhibited) by the heterocyclic compounds of formula I include, but are not limited to: tumors of epithelial origin (adenomas and various types of cancers, including adenocarcinomas, squamous carcinomas, transitional cell carcinomas and other carcinomas) such as bladder and urinary tract cancers, breast cancers, gastrointestinal tract cancers (including esophageal cancers, stomach cancers (stomach), small intestine cancers, colon cancers, rectal cancers and anal cancers), liver cancers (hepatocellular cancers), gall bladder and biliary tract system cancers, exocrine pancreatic cancers, kidneys, lungs (e.g., adenocarcinoma, small cell lung cancer, non-small cell lung cancer, bronchioloalveolar cancers and mesothelioma), head and neck cancers (e.g., tongue cancers, oral cancers, laryngeal cancers, pharyngeal cancers, nasopharyngeal cancers, tonsil cancers, salivary gland cancers, nasal and sinus), reproductive system (e.g., ovary, fallopian tube, peritoneum, vagina, vulva, penis, cervix, myometrium, endometrial cancers, thyroid cancers (e.g., thyroid follicular cancers), adrenal cancers, prostate cancers, skin cancers (e.g., melanoma, basal cell cancers, squamous cell cancers, keratoacanthoma, dysplastic nevi); hematological malignancies (i.e., leukemias, lymphomas) and precancerous hematological and borderline malignant diseases, including hematological malignancies and related lymphoid diseases (e.g., acute lymphoblastic leukemia, chronic lymphoblastic leukemia, B-cell lymphomas, such as diffuse large B-cell lymphomas, follicular lymphomas, burkitt's lymphomas, mantle cell lymphomas, MALT lymphomas, T-cell lymphomas and leukemias, natural killer cell lymphomas, hodgkin's lymphomas, hairy cell leukemias, meaningless monoclonal gammaglobulinases, plasmacytomas, multiple myeloma, and post-transplant lymphoproliferative diseases), as well as hematological malignancies and myeloid-related diseases (e.g., acute myelogenous leukemia, chronic myelomonocytic leukemia, eosinophilic granulomatosis, myeloproliferative diseases, such as polycythemia vera), primary thrombocythemia and primary myelofibrosis, myeloproliferative syndromes, myelodysplastic syndromes and promyelocytic leukemias); tumors of mesenchymal origin, such as sarcomas of soft tissue, bone or cartilage, for example osteosarcoma, fibrosarcoma, chondrosarcoma, rhabdomyosarcoma, leiomyosarcoma, liposarcoma, angiosarcoma, kaposi's sarcoma, ewing's sarcoma, gastrointestinal epithelial sarcoma, malignant epithelial sarcoma histiocytoma and cutaneous fibrosarcoma protuberance; tumors of the central or peripheral nervous system (e.g., astrocytomas, gliomas and glioblastomas, meningiomas, ependymomas, pineal tumors and schwannomas); endocrine tumors (e.g., pituitary tumors, adrenal tumors, islet cell tumors, parathyroid tumors, carcinoid tumors, and medullary thyroid cancers); eye and accessory tumors (e.g., retinoblastoma); germ cell and trophoblastic tumors (e.g., teratomas, seminomas, asexual cell tumors, grape embryo and choriocarcinomas); pediatric and embryonic tumors (e.g., medulloblastoma, neuroblastoma, wilms tumor, and primitive neuroectodermal tumors); or congenital or other aspects of the syndrome that predisposes the patient to malignancy (e.g., pigment dry skin disease).

Preferably, the cancer is lymphoma, rhabdoid tumor, multiple myeloma, uterine cancer, gastric cancer, peripheral nervous system cancer, rhabdomyosarcoma, bone cancer, colorectal cancer, mesothelioma, breast cancer, ovarian cancer, lung cancer, fibroblast cancer, central nervous system cancer, urinary tract cancer, upper respiratory tract cancer, leukemia, kidney cancer, skin cancer, esophageal cancer, and pancreatic cancer.

Additional aspects and advantages of the application will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the application.

Definition of terms:

unless otherwise indicated, the radical and term definitions recited in the specification and claims of the present application, including as examples, exemplary definitions, preferred definitions, definitions recited in tables, definitions of specific compounds in the examples, and the like, may be arbitrarily combined and coupled with each other. Such combinations and combinations of radical definitions and structures of compounds should fall within the scope of the present description.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the claimed subject matter belongs. All patents, patent applications, and publications cited herein are hereby incorporated by reference in their entirety unless otherwise indicated. If there are multiple definitions of terms herein, the definitions of this chapter shall control.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the inventive subject matter. In the present application, the singular is used to include the plural unless specifically stated otherwise. It must be noted that, as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. It should also be noted that the use of "or" means "and/or" unless stated otherwise. Furthermore, the terms "include," as well as other forms, such as "comprising," "including," and "containing," are not limiting.

Conventional methods within the skill of the art, such as mass spectrometry, NMR, IR and UV/VIS spectroscopy, and pharmacological methods are employed unless otherwise indicated. Unless specifically defined otherwise, the terms used herein in the description of analytical chemistry, organic synthetic chemistry, and pharmaceutical chemistry are known in the art. Standard techniques may be used in chemical synthesis, chemical analysis, pharmaceutical preparation, formulation and delivery, and treatment of patients. For example, the reaction and purification can be carried out using the manufacturer's instructions for the kit, or in a manner well known in the art or in accordance with the teachings of the present application. The techniques and methods described above may generally be practiced according to conventional methods well known in the art, based on a number of general and more specific descriptions in the literature cited and discussed in this specification. In this specification, groups and substituents thereof can be selected by one skilled in the art to provide stable moieties and compounds.

When substituents are described by conventional formulas written from left to right, the substituents also include chemically equivalent substituents obtained when writing formulas from right to left. For example, CH ₂ O is equal to OCH ₂ . As used herein,representing the attachment site of the group.

The section headings used herein are for purposes of organizing articles only and should not be construed as limiting the subject matter. All documents or portions of documents cited in this disclosure, including but not limited to patents, patent applications, articles, books, operating manuals, and treatises, are hereby incorporated by reference in their entirety.

In addition to the foregoing, when used in the specification and claims of the present application, the following terms have the meanings indicated below, unless otherwise specified.

Where a range of values recited in the specification and claims is understood to be an "integer," it is understood that both ends of the range and each integer within the range are recited. For example, an "integer of 1 to 6" should be understood to describe each integer of 1, 2, 3, 4, 5, and 6.

In the present application, "Pol θ", "POLQ", "Pol θ polymerase" and "Pol θ protein polymerase" have the same definition.

In the present application, the term "halogen" means fluorine, chlorine, bromine, iodine, alone or as part of other substituents.

As used herein, the term "alkyl" alone or as part of another substituent means a straight or branched hydrocarbon chain group consisting of only carbon and hydrogen atoms, free of unsaturation, having, for example, 1 to 6 carbon atoms, and linked to the remainder of the molecule by a single bond. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, pentyl, isopentyl, neopentyl and hexyl. The alkyl group may be unsubstituted or substituted with one or more suitable substituents. The alkyl group can also be an isotopic isomer of a naturally abundant alkyl group that is enriched in isotopes of carbon and/or hydrogen (i.e., deuterium or tritium).

The term "C", alone or as part of another substituent ₁ -C ₆ Alkyl "is understood to mean a straight-chain or branched saturated monovalent hydrocarbon radical having 1,2, 3, 4, 5 or 6 carbon atoms. The alkyl is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-ethylpropyl, 1, 2-dimethylpropyl, neopentyl, 1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3-dimethylbutyl, 2-dimethylpropyl Methylbutyl, 1-dimethylbutyl, 2, 3-dimethylbutyl, 1, 2-dimethylbutyl, or the like, or isomers thereof. In particular, the radicals have 1,2 or 3 carbon atoms ("C ₁ -C ₃ Alkyl "), such as methyl, ethyl, n-propyl or isopropyl.

"haloalkyl", alone or as part of other substituents, is meant to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms, substituted with one or more halogens. For example, C ₁ -C ₆ Haloalkyl refers to a chain or branched saturated monovalent hydrocarbon radical having 1,2, 3, 4, 5, or 6 carbon atoms wherein 1 or more hydrogen atoms are replaced with halogen. Examples of haloalkyl include, but are not limited to, trifluoromethyl, trichloromethyl, pentafluoroethyl, pentachloroethyl, 2-trifluoroethyl, heptafluoropropyl, and heptachloropropyl.

The term "C", alone or as part of another substituent ₁ -C ₆ Alkoxy "is understood to mean a straight-chain or branched saturated monovalent hydrocarbon radical having 1,2, 3, 4, 5 or 6 carbon atoms and an oxygen atom composition, or C ₁ -C ₆ alkyl-O-C ₁ -C ₆ Alkyl groups are defined herein, and oxygen atoms may be attached to C ₁ -C ₆ Alkyl groups on either a straight chain or a straight chain carbon atom. Including but not limited to: methoxy (CH) ₃ -O-), ethoxy (C) ₂ H ₅ -O-), propoxy (C) ₃ H ₇ -O-), butoxy (C) ₄ H ₉ -O-)。

The term "cycloalkyl" or "carbocyclyl" refers to a cyclic alkyl group. The term "m-n membered cycloalkyl" or "C _m -C _n Cycloalkyl "is understood to mean a saturated, unsaturated or partially saturated carbocyclic ring having m to n atoms. For example, "3-8 membered cycloalkyl" or "C ₃ -C ₈ Cycloalkyl "refers to a cyclic alkyl group containing 3 to 8 carbon atoms. "3-6 membered cycloalkyl" contains 3-6 carbon atoms. Including monocyclic, bicyclic, tricyclic, spiro, or bridged rings. Examples of unsubstituted cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl. Ring(s)The alkyl group may be substituted with one or more substituents.

The term "heterocycle" or "heterocycloalkyl" or "heterocyclyl" when used alone or as part of another substituent means a cycloalkyl group in which one or more (in some embodiments 1 to 3) carbon atoms are replaced with heteroatoms such as, but not limited to N, O, S and P. The term "m-n membered heterocycloalkyl" is understood to mean a saturated, unsaturated or partially saturated ring having m to n atoms. For example, the term "4-10 membered heterocycloalkyl" is understood to mean a saturated, unsaturated or partially saturated ring having 4 to 10 atoms. In some embodiments, the heterocycloalkyl group can be a heterocycloalkyl group fused with an aryl or heteroaryl group. When a prefix such as 3-8 membered is used to represent a heterocycloalkyl group, the number of carbons is also meant to include heteroatoms. Including monocyclic, bicyclic, tricyclic, spiro, or bridged rings. Non-limiting examples of heterocycles include, but are not limited to, dihydropyridazine, dihydropyrazines, including substituted forms thereof, such as heterocyclyl groups including, but not limited to, 6-oxo-1, 6-dihydropyridazin-3-yl, 4-methyl-5-oxo-4, 5-dihydropyrazin-2-yl, and the like.

The term "heteroaryl" or "heteroaryl ring" when used alone or as part of another substituent refers to a monocyclic or polycyclic aromatic ring system in which 1 to 3 atoms in the ring system are heteroatoms, i.e., elements other than carbon, including but not limited to N, O, S or P. Such as furyl, imidazolyl, indolinyl, pyrrolidinyl, pyrimidinyl, tetrazolyl, thienyl, pyridyl, pyrrolyl, N-methylpyrrolyl, quinolinyl and isoquinolinyl. Heteroaryl groups may optionally be fused to a benzene ring and may also be monocyclic, bicyclic, tricyclic, spiro, or bridged.

"5-6 membered heteroaryl ring radical" when used alone or as part of another substituent is understood to mean an aromatic ring radical having 5 to 6 ring atoms and comprising 1 or more heteroatoms independently selected from N, O and S. Preferably 1 to 3-aromatic ring groups independently selected from heteroatoms of N, O and S. In particular, the heteroaryl ring group is selected from thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl and the like.

In the present application, the term "salt" or "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts. The term "pharmaceutically acceptable" is intended to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

By "pharmaceutically acceptable acid addition salt" is meant a salt with an inorganic or organic acid that retains the biological effectiveness of the free base without other side effects. By "pharmaceutically acceptable base addition salt" is meant a salt formed with an inorganic or organic base that is capable of maintaining the bioavailability of the free acid without other side effects. In addition to pharmaceutically acceptable salts, other salts are contemplated by the present application. They may serve as intermediates in the purification of the compounds or in the preparation of other pharmaceutically acceptable salts or may be used in the identification, characterization or purification of the compounds of the application.

The term "amine salt" refers to the product of neutralizing an alkyl primary, secondary or tertiary amine with an acid. The acid includes an inorganic acid or an organic acid as described in the present application.

The term "stereoisomer" refers to an isomer produced by the spatial arrangement of atoms in a molecule, and includes cis-trans isomers, enantiomers, non-corresponding isomers and conformational isomers.

Depending on the choice of starting materials and methods, the compounds according to the invention may be present in the form of one of the possible isomers or mixtures thereof, for example as pure optical isomers or as isomer mixtures, for example as racemic and diastereomeric mixtures, depending on the number of asymmetric carbon atoms. When describing optically active compounds, the prefix D and L or R and S are used to denote the absolute configuration of the molecule in terms of chiral center (or chiral centers) in the molecule. The prefixes D and L or (+) and (-) are symbols for designating the rotation of plane polarized light by a compound, where (-) or L represents that the compound is left-handed. The compound prefixed with (+) or D is dextrorotatory.

When the bonds to chiral carbons in the formulae of the present invention are depicted in straight lines, it is understood that both the (R) and (S) configurations of the chiral carbons and the enantiomerically pure compounds and mixtures thereof resulting therefrom are included within the general formula. The absolute configuration of a solid center is represented by wedge-shaped keys and dashed keys.

In the present application, "pharmaceutical composition" refers to a formulation of a compound of the present application with a medium commonly accepted in the art for delivery of biologically active compounds to a mammal (e.g., a human). The medium includes a pharmaceutically acceptable carrier. The purpose of the pharmaceutical composition is to promote the administration of organisms, facilitate the absorption of active ingredients and further exert biological activity.

In the present application, "pharmaceutically acceptable carrier" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye/colorant, flavoring agent, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonizing agent, solvent, or emulsifying agent that is approved by the relevant government regulatory agency as acceptable for human or livestock use.

The compounds of the present application may contain non-natural proportions of atomic isotopes on one or more of the atoms comprising the compounds. For example, compounds may be labeled with radioisotopes, such as deuterium ² H) Tritium% ³ H) Iodine-125% ¹²⁵ I) Or C-14% ¹⁴ C) A. The application relates to a method for producing a fibre-reinforced plastic composite All isotopic variations of the compounds of the present application, whether radioactive or not, are intended to be encompassed within the scope of the present application.

The term "adjuvant" refers to a pharmaceutically acceptable inert ingredient. Examples of the category of the term "excipient" include, without limitation, binders, disintegrants, lubricants, glidants, stabilizers, fillers, diluents, and the like. Excipients can enhance the handling characteristics of the pharmaceutical formulation, i.e., by increasing flowability and/or tackiness, making the formulation more suitable for direct compression.

The term "treatment" and other similar synonyms as used herein include the following meanings:

(i) Preventing the occurrence of a disease or disorder in a mammal, particularly when such mammal is susceptible to the disease or disorder, but has not been diagnosed as having the disease or disorder;

(ii) Inhibiting the disease or disorder, i.e., inhibiting its progression;

(iii) Alleviating a disease or condition, i.e., causing the state of the disease or condition to subside; or alternatively

(iv) Alleviating symptoms caused by the disease or condition.

The reaction temperature of each step may be appropriately selected depending on the solvent, starting material, reagent, etc., and the reaction time may be appropriately selected depending on the reaction temperature, solvent, starting material, reagent, etc. After the reaction of each step is finished, the target compound can be separated and purified from the reaction system according to a common method, such as filtration, extraction, recrystallization, washing, silica gel column chromatography and the like. Under the condition of not influencing the next reaction, the target compound can also directly enter the next reaction without separation and purification.

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 inventor has studied extensively and intensively, and unexpectedly developed a Pol theta inhibitor which has a structure shown in the formula I, has good inhibition effect on Pol theta polymerase, can prevent or treat diseases or symptoms mediated by Pol theta, shows excellent pharmacokinetic properties, and has higher safety and pharmaceutical properties.

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.

EXAMPLE 1 preparation of Compound I-1A

The synthetic route is as follows:

the first step: preparation of 1- (tert-butyl) 2-ethyl (R) -5-oxopyrrolidine-1, 2-dicarboxylic acid ester (B1-2)

Ethyl (R) -5-oxopyrrolidine-2-carboxylate (10.0 g,63.6 mmol) and di-tert-butyl dicarbonate (16.66 g,76 mmol) were dissolved in tetrahydrofuran (100 mL), N-dimethylpyridine-4-amine (0.777 g,6.36 mmol) was added at 0deg.C, and the reaction stirred at 25deg.C for 10 hours. After completion of the reaction, the reaction mixture was concentrated, then dissolved in water (20 mL), extracted with ethyl acetate (20 mL x 3), the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give crude 1- (tert-butyl) 2-ethyl (R) -5-oxopyrrolidine-1, 2-dicarboxylic acid ester (13.9 g, 85% yield).

LC-MS,M/Z(ESI):258.2[M+H] ⁺

And a second step of: preparation of 1- (tert-butyl) 2-ethyl (R, E) -4- ((dimethylamino) methylene) -5-oxopyrrolidine-1, 2-dicarboxylic acid ester (B1-4)

1- (tert-butyl) 2-ethyl (R) -5-oxopyrrolidine-1, 2-dicarboxylic acid ester (13 g,50.5 mmol) was dissolved in ethylene glycol dimethyl ether (70 mL), and 1-tert-butoxy-N, N, N ', N' -tetramethylmethane diamine (14.09 g,81 mmol) was added thereto, and the reaction solution was refluxed at 100℃for 10 hours. After the completion of the reaction, the reaction solution was cooled to room temperature, and then concentrated in vacuo to give crude 1- (tert-butyl) 2-ethyl (R, E) -4- ((dimethylamino) methylene) -5-oxopyrrolidine-1, 2-dicarboxylic acid ester (13.0 g, yield 82%), which was directly used in the next step.

And a third step of: preparation of 1- (tert-butyl) 2-ethyl (R) -4-methylene-5-oxopyrrolidine-1, 2-dicarboxylic acid ester (B1-5)

1- (tert-butyl) 2-ethyl (R, E) -4- ((dimethylamino) methylene) -5-oxopyrrolidine-1, 2-dicarboxylic acid ester (13.0 g,50.5 mmol) was dissolved in tetrahydrofuran (80 mL), 1N hydrochloric acid (25 mL) was added, and the reaction mixture was stirred at 25℃for 2 hours. After the completion of the reaction, the reaction mixture was allowed to separate naturally, the aqueous phase was extracted with tetrahydrofuran (20 mL) 2 times, the organic phases were combined, and then potassium carbonate (10.13 g,73.3 mmol) and a 37% formaldehyde solution (40 mL) were added to the reaction mixture, and the reaction mixture was stirred at 25℃for 45 minutes. After the reaction was completed, the reaction solution was concentrated, then dissolved in water (20 mL), extracted with ethyl acetate (40 mL x 3), the organic layers were combined, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a crude product. The crude product was purified by column chromatography (silica gel, petroleum ether: ethyl acetate=10:1 to 2:1) to give 1- (tert-butyl) 2-ethyl (R) -4-methylene-5-oxopyrrolidine-1, 2-dicarboxylic acid ester (6.0 g, yield 44.1%).

LC-MS,M/Z(ESI):170.0[M-100] ⁺

¹ H NMR(400MHz,CDCl ₃ )δ6.23(t,J＝2.8Hz,1H),5.51(t,J＝2.4Hz,1H),4.60(dt,J＝8.8,4.4Hz,1H),4.27–4.16(m,2H),3.07(ddt,J＝17.3,10.1,3.0Hz,1H),2.71(ddd,J＝17.5,5.4,2.4Hz,1H),1.51(s,9H),1.31–1.23(m,3H).

Fourth step: preparation of 5- (tert-butyl) 6-ethyl (R) -4-oxo-5-azaspiro [2.4] heptane-5, 6-dicarboxylic acid ester (B1-6)

1- (tert-butyl) 2-ethyl (R) -4-methylene-5-oxopyrrolidine-1, 2-dicarboxylic acid ester (6.0 g,134 mmol) and palladium acetate (0.5 g,2.228 mmol) were dissolved in diethyl ether (10 mL), then replaced three times with nitrogen and vacuum ester was applied to the rubber stopper to prevent leakage. The reaction mixture was cooled to-10℃and fresh diazomethane diethyl ether solution (5.62 g,134 mmol) was slowly added and the reaction mixture was stirred at-10℃for 0.5 hours. Then, the temperature of the reaction solution was raised to 25℃and the reaction solution was stirred at 25℃for 10 hours. After the completion of the reaction, the reaction mixture was filtered through celite, then washed with ethyl acetate, and concentrated to give 5- (t-butyl) 6-ethyl (R) -4-oxo-5-azaspiro [2.4] heptane-5, 6-dicarboxylic acid ester (6.0 g, yield 95%).

LC-MS,M/Z(ESI):184.3[M-100] ⁺

Fifth step: preparation of (R) -4-oxo-5-azaspiro [2.4] heptane-6-carboxylic acid ethyl ester (B1-7)

5- (tert-butyl) 6-ethyl (R) -4-oxo-5-azaspiro [2.4] heptane-5, 6-dicarboxylic acid ester (6.0 g,21.18 mmol) was dissolved in dichloromethane (60.0 mL), hydrochloric acid/ethyl acetate (20 mL, 4M) was added at 0℃and the reaction solution was then warmed to 25℃and stirred at 25℃for 2 hours. After the completion of the reaction, the reaction mixture was directly dried by spin-drying, and concentrated to give crude (R) -4-oxo-5-azaspiro [2.4] heptane-6-carboxylic acid ethyl ester (3.8 g, yield 98%).

LC-MS,M/Z(ESI):184.2[M+H] ⁺

Sixth step: preparation of (R) -4-oxo-5-azaspiro [2.4] heptane-6-carboxylic acid (B1-8)

Ethyl (R) -4-oxo-5-azaspiro [2.4] heptane-6-carboxylate (4.7 g,25.7 mmol) was dissolved in a solution of tetrahydrofuran (27 mL), water (18 mL) and methanol (3 mL), and lithium hydroxide monohydrate (3.23 g,77 mmol) was added thereto, and the reaction was stirred at 25℃for 16 hours. After the reaction was completed, the reaction mixture was concentrated, then extracted with ethyl acetate (20 mL), the impurities were extracted into an organic phase, then the aqueous phase was adjusted to pH 3 with 1N hydrochloric acid, extracted with dichloromethane (30 mL x 5), the organic phases were combined, dried over anhydrous sodium sulfate, and filtered and concentrated to give crude (R) -4-oxo-5-azaspiro [2.4] heptane-6-carboxylic acid (2.4 g, 60.3%).

LC-MS,M/Z(ESI):156.0[M+H] ⁺

¹ H NMR(400MHz,CDCl ₃ )δ10.19(s,1H),7.65(s,1H),4.13(q,J＝7.1Hz,1H),2.08(d,J＝19.5Hz,2H),1.26(t,J＝7.1Hz,2H),1.21–1.15(m,1H),0.91–0.79(m,1H).

Seventh step: preparation of (R) -N- (5-chloro-2, 4-difluorophenyl) -4-oxo-5-azaspiro [2.4] heptane-6-carboxamide (B1-10)

(R) -4-oxo-5-azaspiro [2.4]Heptane-6-carboxylic acid (2.4 g,15.47 mmol) and 5-chloro-2, 4-difluoroaniline (2.78 g,17.02 mmol) were dissolved in N, N-dimethylformamide (25 mL), and 1-propylphosphoric anhydride (T) was added ₃ An ethyl acetate solution of P) (24.61 g,38.7mmol,50% ethyl acetate) and pyridine (6.26 mL,77 mmol), and the reaction was heated to 50deg.C and stirred under this condition for 10 hours. After completion of the reaction, the reaction mixture was dissolved in water (10 mL) and ethyl acetate (20 mL), the organic phase was separated, and the aqueous phase was extracted with ethyl acetate (20 ml×3) The organic phases were combined, washed with brine (20 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by column chromatography (silica gel, petroleum ether: ethyl acetate=10:1 to 5:1) to give (R) -N- (5-chloro-2, 4-difluorophenyl) -4-oxo-5-azaspiro [2.4]Heptane-6-carboxamide (3.6 g, 77% yield).

LC-MS,M/Z(ESI):301.6[M+H] ⁺

Eighth step: preparation of (R) -N- (5-chloro-2, 4-difluorophenyl) -5- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -4-oxo-5-azaspiro [2.4] heptane-6-carboxamide (B1-12)

(R) -N- (5-chloro-2, 4-difluorophenyl) -4-oxo-5-azaspiro [2.4]Heptane-6-carboxamide (1.6 g,5.32 mmol) and 2-chloro-6-methyl-4- (trifluoromethyl) pyridine (1.249 g,6,39 mmol) were dissolved in 1, 4-dioxane (15 mL), followed by the addition of potassium carbonate (2.206 g,15.96 mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (XantPhos, 0.616g,1.064 mmol) and tris (dibenzylideneacetone) dipalladium (Pd ₂ dba ₃ 0.487g,6.39 mmol) and then three times with nitrogen, at 100℃for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature, water (5 mL) and ethyl acetate (10 mL) were added to the reaction solution to dissolve, an organic phase was separated, an aqueous phase was extracted with ethyl acetate (10 ml×3), the organic phases were combined, and the organic phase was washed with brine (20 mL) and dried over anhydrous sodium sulfate, and then concentrated by filtration to obtain a crude product. The crude product is purified by column chromatography (silica gel, ethyl acetate: petroleum ether=8:1 to 3:1) to give (R) -N- (5-chloro-2, 4-difluorophenyl) -5- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -4-oxo-5-azaspiro [2.4 ]Heptane-6-carboxamide (517 mg, 21.13% yield).

LC-MS,M/Z(ESI):460.2[M+H] ⁺

¹ H NMR(400MHz,CDCl ₃ )δ8.94(s,1H),8.59(s,1H),8.45(t,1H),7.14(s,1H),6.95(dd,1H),5.31(s,1H),2.65–2.59(m,1H),2.56(d,3H),2.51(dd,1H),1.43(ddd,J＝10.8,7.0,3.9Hz,1H),1.37–1.30(m,1H),1.20–1.13(m,1H),0.99–0.93(m,1H).

Ninth step: (R) -N- (5-chloro-2, 4-difluorophenyl) -N-methyl-5- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -4-oxo-5-azaspiro [2.4] heptane-6-carboxamide (I-1A)

(R) -N- (5-chloro-2, 4-difluorophenyl) -5- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -4-oxo-5-azaspiro [2.4] heptane-6-carboxamide (517 mg,1.124 mmol) and cesium carbonate (733 mg,2.249 mmol) were dissolved in N, N-dimethylformamide (5 mL), methyl iodide (192 mg,1.349 mmol) was added, and the reaction was warmed to 50℃and stirred at 50℃for 1 hour. After the completion of the reaction, the reaction solution was cooled to room temperature, water (3 mL) and ethyl acetate (5 mL) were added to the reaction solution to dissolve, the organic phase was separated, the aqueous phase was extracted with ethyl acetate (5 ml×3), the organic phases were combined, and the organic phase was washed with brine (5 mL) and dried over anhydrous sodium sulfate, and then concentrated by filtration to give a crude product. The crude product was isolated and purified by reverse phase high performance liquid chromatography to give (R) -N- (5-chloro-2, 4-difluorophenyl) -N-methyl-5- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -4-oxo-5-azaspiro [2.4] heptane-6-carboxamide (357 mg, 67% yield).

LC-MS,M/Z(ESI):473.8[M+H] ⁺

¹ H NMR(400MHz,CDCl ₃ )δ8.64–8.51(m,1H),7.95(t,1H),7.17–7.10(m,1H),7.07(d,1H),5.11(dd,1H),3.27(s,3H),2.65(s,2H),2.53(d,1H),2.22(dd,1H),2.03(dd,1H),1.39–1.34(m,1H),1.31–1.27(m,1H),0.99–0.94(m,1H),0.81(dd,1H).

Preparation of Compound I-1B reference I-1A preparation method

EXAMPLE 2 preparation of Compound I-4

The synthetic route is shown below:

the first step: synthesis of (S) -1- (tert-butoxycarbonyl) -4, 4-difluoropyrrolidine-2-carboxylic acid (B4-2)

To a solution of 1- (tert-butyl) 2-methyl (S) -4, 4-difluoropyrrolidine-1, 2-dicarboxylic acid ester (2.0 g,7.54 mmol) in methanol (20 mL) was added water (2 mL) at room temperature. LiOH H is added into the reaction liquid under stirring ₂ O (92.2 g,365.4 mmol), the reaction was stirred at 25℃for 12h. After the reaction was completed, the reaction solution was concentrated to dryness, 10mL of water was added, and the ph=3 was adjusted with 1M aqueous hydrochloric acid with stirring. The reaction mixture was extracted with ethyl acetate (20 ml x 3), the organic phases were combined, dried over sodium sulfate and concentrated to give crude (S) -1- (tert-butoxycarbonyl) -4, 4-difluoropyrrolidine-2-carboxylic acid (1.4 g, 74% yield).

LC-MS,M/Z(ESI):252.0[M+H] ⁺ 。

And a second step of: synthesis of (S) -2- ((5-chloro-2, 4-difluorophenyl) carbamoyl) -4, 4-difluoropyrrolidine-1-carboxylic acid tert-butyl ester (B4-3)

Into a 100mL round bottom flask were charged (S) -1- (T-butoxycarbonyl) -4, 4-difluoropyrrolidine-2-carboxylic acid (1.4 g,5.6 mmol), 5-chloro-2, 4-difluoroaniline (1.09 g,6.7 mmol) and N, N-dimethylformamide (15 mL), the reaction mixture was cooled to 0-5℃and N, N-diisopropylethylamine (2.16 g,16.7 mmol) was added to the reaction mixture, followed by slow addition of 1-propylphosphoric acid cyclic anhydride (T) ₃ P,4.25g,6.68mmol,50% N, N-dimethylformamide solution), and the reaction was stirred at room temperature for 2 hours. The reaction was quenched by addition of saturated sodium bicarbonate solution (40 mL), then 50mL of ethyl acetate, the organic phase was collected by liquid separation, washed with saturated sodium bicarbonate solution (40 mL x 2), then with saturated NaCl (40 mL) solution, dried over sodium sulfate, concentrated to give crude product, which was purified by silica gel column (ethyl acetate/petroleum ether=1/5) to give (S) -2- ((5-chloro-2, 4-difluorophenyl) carbamoyl) -4, 4-difluoropyrrolidine-1-carboxylic acid tert-butyl ester (1.7 g, yield 77%).

LC-MS,M/Z(ESI):397.1[M+H] ⁺ 。

And a third step of: synthesis of (S) -2- ((5-chloro-2, 4-difluorophenyl) (methyl) carbamoyl) -4, 4-difluoropyrrolidine-1-carboxylic acid tert-butyl ester (B4-4)

(S) -2- ((5-chloro-2, 4-difluorophenyl) carbamoyl) -4, 4-difluoropyrrolidine-1-carboxylic acid tert-butyl ester (1.7 g,4.28 mmol) was dissolved in N, N-dimethylformamide (15 mL), cesium carbonate (2.1 g,6.43 mmol) was added, the reaction solution was stirred at room temperature for 0.5h, meI (0.9 g,6.43 mmol) was added, and the reaction solution was stirred at room temperature for 2h. To the reaction solution were added water (50 mL) and ethyl acetate (50 mL), and the organic phase was collected by separation, washed with water (50 mL x 2), dried over sodium sulfate, and concentrated to give crude (S) -2- ((5-chloro-2, 4-difluorophenyl) (methyl) carbamoyl) -4, 4-difluoropyrrolidine-1-carboxylic acid tert-butyl ester (1.6 g, yield 88%) which was used directly in the next step.

LC-MS,M/Z(ESI):411.1[M+H] ⁺ 。

Fourth step: synthesis of (S) -5- ((5-chloro-2, 4-difluorophenyl) (methyl) carbamoyl) -3, 3-difluoro-2-oxopyrrolidine-1-carboxylic acid tert-butyl ester (B4-5)

Sodium periodate (4.03 g,18.8 mmol) and water (45 mL) were added to a three-necked flask, the reaction system was purged with nitrogen 3 times, and RuCl was added to the reaction system ₃ (0.78 g,3.77 mmol) and stirred at room temperature for 0.5h. To the reaction was slowly added dropwise a solution of (S) -2- ((5-chloro-2, 4-difluorophenyl) (methyl) carbamoyl) -4, 4-difluoropyrrolidine-1-carboxylic acid tert-butyl ester (1.6 g,3.77 mmol) in ethyl acetate (15 mL), and after the addition was completed, the reaction solution was stirred for 16h. Isopropanol (5 mL) was added to the reaction and stirring was continued for 3h at room temperature, ethyl acetate (50 mL) was added, the aqueous phase was extracted with ethyl acetate (10 mL x 2), the organic phases were combined and concentrated to give crude product, which was purified by silica gel column (ethyl acetate/petroleum ether=1/5) to give (S) -5- ((5-chloro-2, 4-difluorophenyl) (methyl) carbamoyl) -3, 3-difluoro-2-oxopyrrolidine-1-carboxylic acid tert-butyl ester (0.15 g, 9.4% yield).

LC-MS,M/Z(ESI):425.1[M+H] ⁺ 。

Fifth step: synthesis of (S) -N- (5-chloro-2, 4-difluorophenyl) -4, 4-difluoro-N-methyl-5-oxopyrrolidine-2-carboxamide (B4-6)

(S) -5- ((5-chloro-2, 4-difluorophenyl) (methyl) carbamoyl) -3, 3-difluoro-2-oxopyrrolidine-1-carboxylic acid tert-butyl ester (0.15 g,0.35 mmol) was dissolved in dichloromethane (3 mL), znBr was added to the reaction solution ₂ (0.32 g,1.41 mmol) and the reaction was stirred at room temperature for 4h. After the reaction is completed, the reaction is carried outSaturated sodium bicarbonate solution (5 mL) was added to the system, the organic phase was collected by separation, washed with saturated sodium bicarbonate solution (5 mL) and water (5 mL), dried over sodium sulfate, filtered, and concentrated to give crude (S) -N- (5-chloro-2, 4-difluorophenyl) -4, 4-difluoro-N-methyl-5-oxopyrrolidine-2-carboxamide (0.11 g, 96% yield) which was used directly in the next step.

LC-MS,M/Z(ESI):325.0[M+H] ⁺ 。

Sixth step: synthesis of (S) -N- (5-chloro-2, 4-difluorophenyl) -4, 4-difluoro-N-methyl-1- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -5-oxopyrrolidine-2-carboxamide (I-4)

(S) -N- (5-chloro-2, 4-difluorophenyl) -4, 4-difluoro-N-methyl-5-oxopyrrolidine-2-carboxamide (0.11 g,0.34 mmol), potassium carbonate (0.093 g,0.68 mmol), tris (dibenzylideneacetone) dipalladium (0.02 g,0.034 mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (0.02 g,0.034 mmol), 2-chloro-6-methyl-4- (trifluoromethyl) pyridine (0.1 g,0.51 mmol) were dissolved in a solution of 1, 4-dioxane (2 mL), the reaction system was replaced with argon three times, and then reacted at 100℃for 12h. The reaction solution was filtered, and the filtrate was concentrated to dryness to give a crude product, which was purified by a silica gel plate to give (S) -N- (5-chloro-2, 4-difluorophenyl) -4, 4-difluoro-N-methyl-1- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -5-oxopyrrolidine-2-carboxamide (I-4) (26 mg, yield 16%).

¹ H NMR(400MHz,DMSO-d6)δ8.41–8.34(m,1H),8.26–7.99(m,1H),7.96–7.74(m,1H),7.71–7.54(m,1H),5.93–4.94(m,1H),3.19–3.10(m,3H),3.01–2.52(m,5H)。

LC-MS,M/Z(ESI):484.1[M+H] ⁺ 。

EXAMPLE 3 preparation of Compound I-5

The synthetic route is shown below:

the first step: synthesis of 1- (tert-butyl) 2-methyl (S) -4- (((trifluoromethyl) sulfonyl) oxy) -2, 5-dihydro-1H-pyrrole-1, 2-dicarboxylic acid ester (B5-2)

1- (tert-butyl) 2-methyl (S) -4-oxopyrrolidine-1, 2-dicarboxylic acid ester (5.0 g,20.54 mmol) was dissolved in tetrahydrofuran (50 mL) under nitrogen and the reaction solution was cooled to-78 ℃. Lithium bis (trimethylsilylamide) (24.6 mL,24.6mmol,1mol/L in tetrahydrofuran) was slowly added to the reaction while stirring, and the reaction solution was stirred at-78℃for 1h. A solution of N-phenylbis (trifluoromethanesulfonyl) imide (9.6 g,24.6 mmol) in tetrahydrofuran (10 mL) was slowly added and the reaction stirred for a further 2h at-78 ℃. To the reaction mixture was added 50mL of saturated sodium bicarbonate to quench the reaction, the mixture was warmed to room temperature, 100mL of ethyl acetate was added, the mixture was separated, the aqueous phase was extracted with ethyl acetate (20 mL of 3), the organic phases were combined, washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, and concentrated to give a crude product, which was purified by silica gel column (ethyl acetate/petroleum ether=1/10, V/V) to give 1- (tert-butyl) 2-methyl (S) -4- (((trifluoromethyl) sulfonyl) oxy) -2, 5-dihydro-1H-pyrrole-1, 2-dicarboxylic acid ester (4.3 g, yield 56%).

And a second step of: synthesis of 1- (tert-butyl) 2-methyl (S) -4-methyl-2, 5-dihydro-1H-pyrrole-1, 2-dicarboxylic acid ester (B5-3)

1- (tert-butyl) 2-methyl (S) -4- (((trifluoromethyl) sulfonyl) oxy) -2, 5-dihydro-1H-pyrrole-1, 2-dicarboxylic acid ester (4.3 g,11.5 mmol), methylboronic acid (2.1 g,34.4 mmol), sodium carbonate (3.64 g,34.4 mmol), and palladium tetrakis triphenylphosphine (1.32 g,1.15 mmol) were dissolved in 1, 4-dioxane (50 mL), argon was replaced three times, and the reaction was carried out overnight at 100 ℃. The reaction solution was filtered and concentrated to give a crude product, which was purified by a silica gel column (ethyl acetate/petroleum ether=1/5, V/V) to give 1- (tert-butyl) 2-methyl (S) -4-methyl-2, 5-dihydro-1H-pyrrole-1, 2-dicarboxylic acid ester (0.4 g, yield 14.5%).

LC-MS,M/Z(ESI):242.1[M+H] ⁺ 。

And a third step of: synthesis of (S) -1- (tert-butoxycarbonyl) -4-methyl-2, 5-dihydro-1H-pyrrole-2-carboxylic acid (B5-4)

1- (tert-butyl) 2-methyl (S) -4-methyl-2, 5-dihydro-1H-pyrrole-1 at room temperature,to a solution of 2-dicarboxylic acid ester (0.4 g,1.66 mmol) in methanol (4 mL) was added water (0.4 mL). LiOH H is added to the reaction under stirring ₂ O (139 mg,3.32 mmol), the reaction was stirred at 25 ℃ for 12h, after the reaction was complete, the reaction was concentrated to dryness, 4mL of water was added, and the ph=3 was adjusted with 1M aqueous hydrochloric acid under stirring. The reaction mixture was extracted with ethyl acetate (5 ml x 3), the organic phases were combined, dried over sodium sulfate and concentrated to give crude (S) -1- (tert-butoxycarbonyl) -4-methyl-2, 5-dihydro-1H-pyrrole-2-carboxylic acid (0.32 g, 85% yield).

LC-MS,M/Z(ESI):228.1[M+H] ⁺ 。

Fourth step: synthesis of (S) -2- ((5-chloro-2, 4-difluorophenyl) carbamoyl) -4-methyl-2, 5-dihydro-1H-pyrrole-1-carboxylic acid tert-butyl ester (B5-5)

To a 50mL round bottom flask was added (S) -1- (tert-butoxycarbonyl) -4-methyl-2, 5-dihydro-1H-pyrrole-2-carboxylic acid (0.32 g,1.4 mmol), 5-chloro-2, 4-difluoroaniline (276 mg,1.7 mmol) and acetonitrile (3 mL). N-methylimidazole (277 mg,4.22 mmol) was added to the reaction, followed by N, N, N ', N' -tetramethyl chloroformidine hexafluorophosphate (477 mg,1.7 mmol), and the reaction was stirred at room temperature for 2h. To the reaction was added saturated sodium bicarbonate solution (10 mL), then 50mL ethyl acetate was added, the organic phase was collected by liquid separation, washed with saturated sodium bicarbonate solution (10 mL x 2), then with saturated NaCl (10 mL) solution, dried over sodium sulfate, concentrated to give crude product, which was purified by silica gel column (ethyl acetate/petroleum ether=1/5) to give (S) -2- ((5-chloro-2, 4-difluorophenyl) carbamoyl) -4-methyl-2, 5-dihydro-1H-pyrrole-1-carboxylic acid tert-butyl ester (0.42 g, 82% yield).

LC-MS,M/Z(ESI):373.1[M+H] ⁺

Fifth step: synthesis of (2S) -2- ((5-chloro-2, 4-difluorophenyl) carbamoyl) -3, 4-dihydroxy-4-methylpyrrolidine-1-carboxylic acid tert-butyl ester (B5-6)

Potassium carbonate (467.1 mg,3.38 mmol), potassium ferricyanide (1.18 mg,3.38 mmol), methanesulfonamide (107 mg,1.13 mmol), potassium osmium dihydrate (17.5 mg,0.056 mmol) and quinine 1,4- (2, 3-naphthyridine) diether (86 mg,0.11 mmol) were added to water (3 mL) and tert-butanol (1.5 mL) under nitrogen. The reaction solution was cooled to 0-5℃and stirred at 0-5℃for 0.5H, followed by slow addition of a solution of (S) -2- ((5-chloro-2, 4-difluorophenyl) carbamoyl) -4-methyl-2, 5-dihydro-1H-pyrrole-1-carboxylic acid tert-butyl ester (0.42 g,1.13 mmol) in tert-butanol (1.5 mL) and stirring at 0-5℃for a further 12H. Ethyl acetate (10 mL) was added to the reaction, the organic phase was collected by liquid separation, washed with saturated NaCl (10 mL) solution, dried over anhydrous sodium sulfate, concentrated to give crude product, which was purified by silica gel column (ethyl acetate/petroleum ether=1/1) to give tert-butyl (2S) -2- ((5-chloro-2, 4-difluorophenyl) carbamoyl) -3, 4-dihydroxy-4-methylpyrrolidine-1-carboxylate (0.24 g, 52% yield).

LC-MS,M/Z(ESI):407.1[M+H] ⁺

Sixth step: synthesis of tert-butyl (3 aR,6S,6 aS) -6- ((5-chloro-2, 4-difluorophenyl) carbamoyl) -2, 3 a-trimethyldihydro-3 aH- [1,3] dioxol [4,5-c ] pyrrole-5 (4H) -carboxylate (B5-7)

(2S) -2- ((5-chloro-2, 4-difluorophenyl) carbamoyl) -3, 4-dihydroxy-4-methylpyrrolidine-1-carboxylic acid tert-butyl ester (0.24 g,0.59 mmol), pyridine 4-methylbenzenesulfonate (37 mg,0.15 mmol), 2-dimethoxypropane (307 mg,2.95 mmol) were added to toluene (3 mL) under nitrogen. The reaction solution was stirred at 100℃for 2h. The reaction solution was then concentrated to give a crude product, which was purified by silica gel column (ethyl acetate/petroleum ether=1/3) to give tert-butyl (3 ar,6s,6 as) -6- ((5-chloro-2, 4-difluorophenyl) carbamoyl) -2, 3 a-trimethyldihydro-3 aH- [1,3] dioxolan [4,5-c ] pyrrole-5 (4H) -carboxylate (0.17 g, yield 64%).

LC-MS,M/Z(ESI):447.1[M+H] ⁺

Seventh step: synthesis of tert-butyl (3 aR,6S,6 aS) -6- ((5-chloro-2, 4-difluorophenyl) (methyl) carbamoyl) -2, 3 a-trimethyldihydro-3 aH- [1,3] dioxol [4,5-c ] pyrrole-5 (4H) -carboxylate (B5-8)

(3 aR,6S,6 aS) -6- ((5-chloro-2, 4-difluorophenyl) carbamoyl) -2, 3 a-trimethyldihydro-3 aH- [1,3] dioxolo [4,5-c ] pyrrole-5 (4H) -carboxylic acid tert-butyl ester (0.17 g,0.38 mmol) was dissolved in N, N-dimethylformamide (2 mL), cesium carbonate (186 mg,0.57 mmol) was added, the reaction solution was stirred at room temperature for 0.5H, meI (81 mg,0.57 mmol) was added, and the reaction solution was continued to be stirred at room temperature for 2H. To the reaction solution were added water (5 mL) and ethyl acetate (5 mL), and the organic phase was collected by separation, washed with water (5 mL x 3), dried over sodium sulfate, and concentrated to give crude (3 ar,6s,6 as) -6- ((5-chloro-2, 4-difluorophenyl) (methyl) carbamoyl) -2, 3 a-trimethyldihydro-3 aH- [1,3] dioxolan [4,5-c ] pyrrole-5 (4H) -carboxylic acid tert-butyl ester (160 mg, 91% yield) which was used directly in the next step.

LC-MS,M/Z(ESI):461.1[M+H] ⁺

Eighth step: synthesis of tert-butyl (3 aS,6S,6 aS) -6- ((5-chloro-2, 4-difluorophenyl (methyl) carbamoyl) -2, 3 a-trimethyl-4-oxodihydro-3 aH- [1,3] dioxolane [4,5-c ] pyrrole-5 (4H) -carboxylate (B5-9)

Into a three-necked flask, sodium periodate (223 mg,1.04 mmol) and water (2 mL) were charged, the reaction system was purged with nitrogen 3 times, and RuO was added to the reaction system ₂ ·xH ₂ O (5.2 mg,0.035 mmol) and stirred at room temperature for 0.5h. (3 aR,6S,6 aS) -6- ((5-chloro-2, 4-difluorophenyl) (methyl) carbamoyl) -2, 3 a-trimethyldihydro-3 aH- [1,3 were slowly added dropwise to the reaction]Dioxolane [4,5-c ]]A solution of pyrrole-5 (4H) -carboxylic acid tert-butyl ester (0.16 g,0.35 mmol) in ethyl acetate (2 mL) was added and the reaction stirred for 16H. Isopropanol (1 mL) was added to the reaction solution and stirring was continued at room temperature for 3h, ethyl acetate (5 mL) was added, the aqueous phase was extracted with ethyl acetate (5 mL x 2), the organic phases were combined and concentrated to give crude product, which was purified on a silica gel column (ethyl acetate/petroleum ether=1/3) to give (3 as,6s,6 as) -6- ((5-chloro-2, 4-difluorophenyl (methyl) carbamoyl) -2, 3 a-trimethyl-4-oxo dihydro-3 aH- [1,3]Dioxolane [4,5-c ]]Pyrrole-5 (4H) -carboxylic acid tert-butyl ester (0.11 g, 67% yield).

LC-MS,M/Z(ESI):475.1[M+H] ⁺

Ninth step: synthesis of (3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -N,2, 6 a-tetramethyl-6-oxotetrahydro-3 aH- [1,3] dioxolane [4,5-c ] pyrrole-4-carboxamide (B5-10)

(3 aS,6S,6 aS) -6- ((5-chloro-2, 4-difluorophenyl (methyl) carbamoyl) -2, 3 a-trimethyl-4-oxodihydro-3 aH- [1, 3)]Dioxolane [4,5-c ]]Pyrrole-5 (4H) -carboxylic acid tert-butyl ester (0.11 g,0.23 mmol) was dissolved in dichloromethane (2 mL), znBr was added to the reaction solution ₂ (209 mg,0.93 mmol) and the reaction was stirred at room temperature for 4h. LC-MS showed complete reaction, adding saturated sodium bicarbonate solution (5 mL) to the reaction system, separating the solution, collecting the organic phase, washing the organic phase with saturated sodium bicarbonate solution (5 mL) and water (5 mL), drying over sodium sulfate, filtering, concentrating to obtain crude (3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -N,2, 6 a-tetramethyl-6-oxotetrahydro-3 aH- [1,3]]Dioxolane [4,5-c ]]Pyrrole-4-carboxamide (90 mg, 100% yield) was used directly in the next step.

LC-MS,M/Z(ESI):375.1[M+H] ⁺

Tenth step: synthesis of (3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -N,2, 6 a-tetramethyl-5- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -6-oxotetrahydro-3 aH- [1,3] dioxolan [4,5-c ] pyrrole-4-carboxamide (B5-11)

(3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -N,2, 6 a-tetramethyl-6-oxotetrahydro-3 aH- [1,3] dioxolane [4,5-c ] pyrrole-4-carboxamide (90 mg,0.24 mmol), potassium carbonate (68 mg,0.48 mmol), tris (dibenzylideneacetone) dipalladium (22 mg,0.024 mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (14 mg,0.024 mmol), 2-chloro-6-methyl-4- (trifluoromethyl) pyridine (70 mg,0.36 mmol) were dissolved in a solution of 1, 4-dioxane (2 mL), the reaction system was replaced with argon three times, and then reacted at 100℃for 12h. The reaction solution was filtered, and the filtrate was concentrated to dryness to give a crude product, which was purified by silica gel column (ethyl acetate/petroleum ether=1/3) to give (3 as,4s,6 as) -N- (5-chloro-2, 4-difluorophenyl) -N,2, 6 a-tetramethyl-5- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -6-oxotetrahydro-3 aH- [1,3] dioxolan [4,5-c ] pyrrole-4-carboxamide (100 mg, yield 78%).

LC-MS,M/Z(ESI):534.1[M+H] ⁺

Eleventh step: (2S, 3S, 4S) -N- (5-chloro-2, 4-difluorophenyl) -3, 4-dihydroxy-N, 4-dimethyl-1- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -5-oxopyrrolidine-2-carboxamide (I-5)

Under the protection of nitrogen, the (3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluoroPhenyl) -N,2, 6 a-tetramethyl-5- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -6-oxotetrahydro-3 aH- [1,3]Dioxolane [4,5-c ]]Pyrrole-4-carboxamide (0.1 g,0.187 mmol) was dissolved in dichloromethane (2 mL) and BCl was then added ₃ Is a solution in methylene chloride (0.38 mL,0.38mmol,1 mol/L). The reaction mixture was stirred at room temperature for 2h, saturated sodium bicarbonate solution (2 mL) was added to quench the reaction, 5mL of methylene chloride was added, the organic phase was collected by separating the liquid, washed with saturated sodium bicarbonate solution (5 mL) and water (5 mL), dried over anhydrous sodium sulfate, and concentrated to give a crude product, which was purified by silica gel plate (ethyl acetate/petroleum ether) to give (2S, 3S, 4S) -N- (5-chloro-2, 4-difluorophenyl) -3, 4-dihydroxy-N, 4-dimethyl-1- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -5-oxopyrrolidine-2-carboxamide (I-5) (36 mg, yield 39%).

¹ H NMR(400MHz,Methanol-d4)δ8.57–8.39(m,1H),8.25–8.01(m,1H),7.60–7.17(m,2H),6.00–5.08(m,1H),4.53–3.92(m,1H),3.81–3.13(m,3H),2.85–2.43(m,3H),1.74–0.93(m,3H).

LC-MS,M/Z(ESI):494.1[M+H] ⁺

EXAMPLE 4 preparation of Compound I-10

The synthetic route is shown below:

the first step: (2S) -1- (tert-Butoxycarbonyl) -4-hydroxy-4-methylpyrrolidine-2-carboxylic acid (B10-2)

(2S)-1-(tert-butoxycarbonyl)-4-hydroxy-4-methylpyrrolidine-2-carboxylic acid

Tert-butyl (5-chloro-2, 4-difluorophenyl) aminomethyl ester (B10-1, 10.0g,43.6 mmol) was dissolved in tetrahydrofuran (110 mL) under nitrogen, then methyl magnesium bromide (3M, 43.62 mL) was slowly added at-20℃and the reaction stirred at 25℃for 1 hour. After the reaction was completed, 200mL of saturated hydrochloric acid (1M) solution was slowly added dropwise thereto at 0 ℃ to quench, the pH was adjusted to 3 to 4, then extracted with dichloromethane (200 mL x 3), the organic phase was washed with saturated sodium chloride solution (200 mL), dried over anhydrous sodium sulfate, and concentrated by filtration to give (2S) -1- (tert-butoxycarbonyl) -4-hydroxy-4-methylpyrrolidine-2-carboxylic acid (B10-2, 11.0g, crude).

And a second step of: 2-benzyl 1- (tert-butyl) (2S, 4S) -4-hydroxy-4-methylpyrrolidine-1, 2-dicarboxyl ester (B10-3)

2-benzyl 1-(tert-butyl)(2S,4S)-4-hydroxy-4-methylpyrrolidine-1,2-dicarboxylate

(2S) -1- (t-Butoxycarbonyl) -4-hydroxy-4-methylpyrrolidine-2-carboxylic acid (B10-2, 10.0g,43.6 mmol) was dissolved in N, N-dimethylformamide (110 mL) under nitrogen, and cesium carbonate (3 eq) and benzyl bromide (4 eq) were slowly added at 0deg.C and the reaction stirred at 25deg.C for 12 hours. After the reaction was completed, water (300 mL) was added to quench, followed by extraction with ethyl acetate (100 ml×3), and the organic phase was washed with saturated sodium chloride solution (800 mL) and dried over anhydrous sodium sulfate, and then concentrated by filtration to give a crude product, which was separated and purified by reverse phase high performance liquid chromatography to give 2-benzyl 1- (tert-butyl) (2 s,4 s) -4-hydroxy-4-methylpyrrolidine-1, 2-dicarboxyl ester (B10-3, 4.00g, yield 26.6%).

LCMS,M/Z(ESI):236.1[M+H] ⁺

And a third step of: (2S, 4S) -1- (tert-Butoxycarbonyl) -4-hydroxy-4-methylpyrrolidine-2-carboxylic acid (B10-4)

(2S,4S)-1-(tert-butoxycarbonyl)-4-hydroxy-4-methylpyrrolidine-2-carboxylic acid

2-benzyl 1- (tert-butyl) (2S, 4S) -4-hydroxy-4-methylpyrrolidine-1, 2-dicarboxyl ester (B10-3, 1.00g,288 mmol) and palladium hydroxide on carbon (10% Wt,0.60 g) were dissolved in ethanol (30.0 mL), replaced with nitrogen three times, and replaced with hydrogen three times. The reaction solution was stirred under a pressure of hydrogen (50 Psi) at 25℃for 12 hours. After completion of the reaction, the reaction solution was concentrated by filtration to give crude (2S, 4S) -1- (t-butoxycarbonyl) -4-hydroxy-4-methylpyrrolidine-2-carboxylic acid (B10-4, 1.00 g) which was used directly in the next step.

Fourth step: tert-butyl (2S, 4S) -2- ((5-chloro-2, 4-difluorophenyl) aminocarbonyl) -4-hydroxy-4-methylpyrrolidine-1-carboxylic acid ester (B10-5)

tert-butyl(2S,4S)-2-((5-chloro-2,4-difluorophenyl)carbamoyl)-4-hydroxy-4-methylpyrrolidine-1-carboxylate

(2S, 4S) -1- (tert-Butoxycarbonyl) -4-hydroxy-4-methylpyrrolidine-2-carboxylic acid (B10-4, 400mg,1.63 mmol) and 5-chloro-2, 4-difluoroaniline (240 mg,1.47 mmol) were dissolved in acetonitrile (5.00 mL), N-methylimidazole (401 mg,4.89 mmol) and N, N, N, N-tetramethyl chloroformidine hexafluorophosphoric acid (282 mg,2.61 mmol) were added successively, and the reaction was stirred at 80℃for 12 hours. After the reaction was completed, the reaction solution was directly concentrated under reduced pressure to give a crude product, which was separated by column chromatography (petroleum ether: ethyl acetate=20:1-5:1 as mobile phase) to give tert-butyl (2 s,4 s) -2- ((5-chloro-2, 4-difluorophenyl) aminocarbonyl) -4-hydroxy-4-methylpyrrolidine-1-carboxylate (B10-5, 200mg, yield 31.4%).

LCMS,M/Z(ESI):291.0[M+H] ⁺

Fifth step: tert-butyl (2S, 4S) -2- ((5-chloro-2, 4-difluorophenyl) (methyl) aminocarbonyl) -4-hydroxy-4-methylpyrrolidine-1-carboxylic acid ester (B10-6)

tert-butyl(2S,4S)-2-((5-chloro-2,4-difluorophenyl)(methyl)carbamoyl)-4-hydroxy-4-methylpyrrolidine-1-carboxylate

(2S, 4S) -2- ((5-chloro-2, 4-difluorophenyl) aminocarbonyl) -4-hydroxy-4-methylpyrrolidine-1-carboxylate (B10-5, 100mg, 255. Mu. Mol) and cesium carbonate (166 mg, 511. Mu. Mol) were dissolved in N, N-dimethylformamide (1.00 mL), and after stirring the reaction solution at 25℃for 30 minutes, methyl iodide (145 mg,1.02 mmol) was slowly added dropwise. The reaction solution was stirred at 25℃for 2 hours. After completion of the reaction, the reaction was quenched with water (10 mL), then extracted with ethyl acetate (10.0 mL x 3) and the organic phase was washed with saturated sodium chloride solution (20.0 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give tert-butyl (2 s,4 s) -2- ((5-chloro-2, 4-difluorophenyl) (methyl) aminocarbonyl) -4-hydroxy-4-methylpyrrolidine-1-carboxylate (B10-6, 150mg, crude).

LCMS,M/Z(ESI):427.3[M+Na] ⁺

Sixth step: tert-butyl (3S, 5S) -5- ((5-chloro-2, 4-difluorophenyl) (methyl) aminocarbonyl) -3-hydroxy-3-methyl-2-oxopyrrolidine-1-carboxylic acid ester (B10-7)

tert-butyl(3S,5S)-5-((5-chloro-2,4-difluorophenyl)(methyl)carbamoyl)-3-hydroxy-3-methyl-2-oxopyrrolidine-1-carboxylate

Ruthenium trichloride hydrate (48.4 mg, 185. Mu. Mol) and sodium periodate (237 mg,1.11 mmol) were dissolved in water (0.5 mL) under nitrogen atmosphere, the reaction solution was stirred at 25℃for 30 minutes by nitrogen substitution three times, then tert-butyl (2S, 4S) -2- ((5-chloro-2, 4-difluorophenyl) (methyl) aminocarbonyl) -4-hydroxy-4-methylpyrrolidine-1-carboxylate (B10-6, 150mg,370. Mu. Mol) dissolved in ethyl acetate (0.5 mL) was slowly added to the reaction solution, and finally the reaction solution was stirred at 25℃for 12 hours. After completion of the reaction, the reaction was quenched by addition of saturated sodium sulfite (10.0 mL) and then extracted with ethyl acetate (10 mL x 3), and the organic phase was washed with saturated sodium chloride solution (20 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give tert-butyl (3 s,5 s) -5- ((5-chloro-2, 4-difluorophenyl) (methyl) aminocarbonyl) -3-hydroxy-3-methyl-2-oxopyrrolidine-1-carboxylate (B10-7, 50.0mg, crude).

LC-MS,M/Z(ESI):419.1[M+H] ⁺

Seventh step: (2S, 4S) -N- (5-chloro-2, 4-difluorophenyl) -4-hydroxy-N, 4-dimethyl-5-oxopyrrolidine-2-carboxamide (B10-8)

(2S,4S)-N-(5-chloro-2,4-difluorophenyl)-4-hydroxy-N,4-dimethyl-5-oxopyrrolidine-2-carboxamide

Tert-butyl (3 s,5 s) -5- ((5-chloro-2, 4-difluorophenyl) (methyl) aminocarbonyl) -3-hydroxy-3-methyl-2-oxopyrrolidine-1-carboxylic acid ester (B10-7, 50.0mg,119 μmol) was dissolved in dichloromethane (1.00 mL), then zinc bromide (134 mg,596 μmol) was added and the reaction stirred at 25 ℃ for 4 hours. After completion of the reaction, the reaction was quenched with saturated sodium bicarbonate (10 mL), then extracted with dichloromethane (5 mL x 3), and the organic phase was washed with saturated sodium chloride solution (5 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give (2 s,4 s) -N- (5-chloro-2, 4-difluorophenyl) -4-hydroxy-N, 4-dimethyl-5-oxopyrrolidine-2-carboxamide (B10-8, 22.0mg, crude).

LC-MS,M/Z(ESI):319.2[M+H] ⁺

Eighth step: (2S, 4S) -N- (5-chloro-2, 4-difluorophenyl) -4-hydroxy-N, 4-dimethyl-1- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -5-oxopyrrolidine-2-carboxamide (I-10)

(2S,4S)-N-(5-chloro-2,4-difluorophenyl)-4-hydroxy-N,4-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-carboxamide

(2S, 4S) -N- (5-chloro-2, 4-difluorophenyl) -4-hydroxy-N, 4-dimethyl-5-oxopyrrolidine-2-carboxamide (B10-8, 20mg, 62.7. Mu. Mol) and 2-chloro-6-methyl-4- (trifluoromethyl) pyridine (24.5 mg,125. Mu. Mol) were dissolved in 1, 4-dioxane (1.00 mL) and cesium carbonate (40.8 mg, 125.51. Mu. Mmol), tris (dibenzylideneacetone) dipalladium (8.62 mg, 9.41. Mu. Mol) and 4, 5-bis (diphenyl) were then added, respectively, at room temperature under nitrogen protection After three nitrogen substitutions of phosphorus-based) -9, 9-dimethylxanthene (10.8 mg, 18.8. Mu. Mol), the reaction mixture was stirred at 100℃for 12 hours. After the reaction is finished, directly filtering and concentrating to obtain brown oily matter, separating and purifying crude product by reverse-phase high-performance liquid chromatography (separation method is that Waters Xbridge 150 x 25mm x 5um, mobile phase: [ water (NH) ₄ HCO ₃ ) -acetonitrile]The method comprises the steps of carrying out a first treatment on the surface of the Gradient 48% -78% B over 9 min), purification gives (2S, 4S) -N- (5-chloro-2, 4-difluorophenyl) -4-hydroxy-N, 4-dimethyl-1- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -5-oxopyrrolidine-2-carboxamide (I-10, 1.60mg, yield 2.64%).

¹ H NMR(400MHz,CH ₃ OH-d4)δ＝8.47-8.56(m,1H),7.84-8.06(m,1),7.50-7.56(m,1H),7.27-7.34(m,1H),4.60-4.96(m,1H),3.26-.3.70(m,1H),2.54-2.78(m,3H),2.04-2.30(m,2H),1.33-1.51(m,3H).

EXAMPLE 5 preparation of Compound I-11

The synthetic route is as follows:

the first step: 1- (tert-butyl) 2-methyl (S) -4-cyclopropyl-2, 5-dihydro-1H-pyrrole-1, 2-dicarboxyl ester (B11-2)

1-(tert-butyl)2-methyl(S)-4-cyclopropyl-2,5-dihydro-1H-pyrrole-1,2-dicarboxylate

(S) -1-tert-butyl 2-methyl 4- (((trifluoromethyl) sulfonyl) oxo) -1H-pyrrole-1, 2 (2H, 5H) -dicarboxyl ester (B11-1, 15.0g,39.9 mmol), cyclopropylboronic acid (10.3 g,119 mmol) and sodium carbonate (12.7 g,119 mmol) were dissolved in 1, 4-dioxane (150 mL), replaced three times with nitrogen, and tetrakis triphenylphosphine palladium (4.62 g,4.00 mmol) was added under nitrogen protection and the reaction stirred at 100deg.C for 12 hours. After the reaction is completed, the crude product is obtained by direct filtration and concentration. The crude product was isolated and purified by column chromatography (petroleum ether: ethyl acetate=10:1 to 3:1 as mobile phase) to give 1- (tert-butyl) 2-methyl (S) -4-cyclopropyl-2, 5-dihydro-1H-pyrrole-1, 2-dicarboxyl ester (B11-2, 3.70g, 37.6% yield).

¹ H NMR(400MHz,CHCl ₃ -d)δ5.22(d,J＝5.38Hz,1H),4.86-5.09(m,1H),3.96-4.01(m,1H),3.61-3.78(m,3H),1.39-1.48(m,10H),0.69-0.82(m,2H),0.42-0.63(m,2H).

And a second step of: (S) -1- (tert-Butoxycarbonyl) -4-cyclopropyl-2, 5-dihydro-1H-pyrrole-2-carboxylic acid (B11-3)

(S)-1-(tert-butoxycarbonyl)-4-cyclopropyl-2,5-dihydro-1H-pyrrole-2-carboxylic acid

1- (tert-butyl) 2-methyl (S) -4-cyclopropyl-2, 5-dihydro-1H-pyrrole-1, 2-dicarboxyl ester (B11-2, 3.70g,13.8 mmol) was dissolved in absolute ethanol (18.5 mL), tetrahydrofuran (37.0 mL) and water (18.5 mL), lithium hydroxide monohydrate (5.81 g,138 mmol) was added, and the reaction mixture was stirred at 25℃for 12 hours. After the reaction was completed, diluted hydrochloric acid (1M) was added to adjust the pH to 3 to 4, followed by extraction with ethyl acetate (50 mL x 3), and the organic phase was washed with saturated sodium chloride solution (50 mL) and dried over anhydrous sodium sulfate, and then filtered and concentrated to give crude (S) -1- (tert-butoxycarbonyl) -4-cyclopropyl-2, 5-dihydro-1H-pyrrole-2-carboxylic acid (B11-3, 3.20g, yield 91.3%) which was directly used in the next step.

And a third step of: tert-butyl (S) -2- ((5-chloro-2, 4-difluorophenyl) aminocarbonyl) -4-cyclopropyl-2, 5-dihydro-1H-pyrrole-1-carboxylic acid ester (B11-4)

tert-butyl(S)-2-((5-chloro-2,4-difluorophenyl)carbamoyl)-4-cyclopropyl-2,5-dihydro-1H-pyrrole-1-carboxylate

(S) -1- (tert-Butoxycarbonyl) -4-cyclopropyl-2, 5-dihydro-1H-pyrrole-2-carboxylic acid (4.70 g,20.6 mmol) and 5-chloro-2, 4-difluoroaniline (1.72 g,10.5 mmol) were dissolved in acetonitrile (30.0 mL), N, N, N-tetramethyl chloroformyl amidine hexafluorophosphoric acid (3.54 g,12.6 mmol) and N-methylimidazole (2.59 g,31.5 mmol) were added respectively, and the reaction was stirred at 25℃for 12 hours. After the reaction is completed, the crude product is obtained by direct filtration and concentration. The crude product was purified by column chromatography (petroleum ether: ethyl acetate=10:1 to 5:1) to give tert-butyl (S) -2- ((5-chloro-2, 4-difluorophenyl) aminocarbonyl) -4-cyclopropyl-2, 5-dihydro-1H-pyrrole-1-carboxylate (3.80 g, yield: 90.5%).

¹ H NMR(400MHz,CHCl ₃ -d)δ＝8.44(br s,1H),6.95(br s,1H),5.46-5.53(m,1H),4.93-5.18(m,2H),1.38-1.51(m,10H),0.80-0.81(m,2H),0.76-0.78(m,2H).

Fourth step: tert-butyl (2S) -2- ((5-chloro-2, 4-difluorophenyl) aminocarbonyl) -4-cyclopropyl-3, 4-dihydroxypyrrolidine-1-carboxylic acid ester (B11-5)

tert-butyl(2S)-2-((5-chloro-2,4-difluorophenyl)carbamoyl)-4-cyclopropyl-3,4-dihydroxypyrrolidine-1-carboxylate

Methanesulfonamide (1.02 g,7.02 mmol), potassium carbonate (2.91 g,21.06 mmol), potassium ferricyanide (6.93 g,21.06 mmol), hydroquinine 1,4- (2, 3-naphthyridine) diether (546.88 mg, 702.05. Mu. Mol), and potassium osmium dihydrate (129.34 mg, 351.03. Mu. Mol) were dissolved in t-butanol (5.00 mL) and water (5.00 mL), the reaction solution was stirred at 0℃for 30 minutes after nitrogen substitution three times, and then t-butyl (S) -2- ((5-chloro-2, 4-difluorophenyl) aminocarbonyl) -4-cyclopropyl-2, 5-dihydro-1H-pyrrole-1-carboxylic acid ester (B11-4, 2.8g,7.02 mmol) dissolved in t-butanol (5.00 mL) was slowly added to the reaction solution at 0℃and the reaction solution was stirred at 0℃for 12 hours. After completion of the reaction, the reaction mixture was filtered and extracted with ethyl acetate (20 ml×3), and the organic phase was washed with saturated sodium chloride solution (40 mL) and dried over anhydrous sodium sulfate, and concentrated by filtration to give a crude product. The crude product was isolated and purified by column chromatography (petroleum ether: ethyl acetate=10:1-2:1) to give tert-butyl (2S) -2- ((5-chloro-2, 4-difluorophenyl) aminocarbonyl) -4-cyclopropyl-3, 4-dihydroxypyrrolidine-1-carboxylate (B11-5, 600mg, 19.7% yield).

LC-MS,M/Z(ESI):307.2(M+H) ⁺

Fifth step: tert-butyl (3 ar,6s,6 as) -6- ((5-chloro-2, 4-difluorophenyl) aminocarbonyl) -3 a-cyclopropyl-2, 2-dimethyltetrahydro-5H- [1,3] dioxolo [4,5-c ] pyrrole-5-carboxylic acid ester (B11-6)

tert-butyl(3aR,6S,6aS)-6-((5-chloro-2,4-difluorophenyl)carbamoyl)-3a-cyclopropyl-2,2-dimethyltetrahydro-5H-[1,3]dioxolo[4,5-c]pyrrole-5-carboxylate

Tert-butyl (2S) -2- ((5-chloro-2, 4-difluorophenyl) aminocarbonyl) -4-cyclopropyl-3, 4-dihydroxypyrrolidine-1-carboxylic acid ester (B11-5, 435mg, 973. Mu. Mol) and 2, 2-dimethoxypropane (506 mg,4.87 mmol) were dissolved in toluene (4.00 mL), and pyridine p-toluenesulfonate (61.1 mg, 243. Mu. Mol) was added, and the reaction was stirred at 100℃for 2 hours. After the reaction is completed, the crude product is obtained by direct concentration. The crude product was purified by column chromatography (petroleum ether: ethyl acetate=10:1-5:1 as mobile phase) to give tert-butyl (3 ar,6s,6 as) -6- ((5-chloro-2, 4-difluorophenyl) aminocarbonyl) -3 a-cyclopropyl-2, 2-dimethyltetrahydro-5H- [1,3] dioxolo [4,5-c ] pyrrole-5-carboxylate (B11-6, 230mg, 46.8% yield).

¹ H NMR(400MHz,CHCl ₃ -d)δ＝7.58-7.68(m,1H),7.07-7.27(m,1H),4.12–4.22(m,1H),3.65-3.82(m,1H),3.42-3.67(m,1H),3.36-3.40(m,1H),3.24(s,3H)1.39–1.50(m,15H),1.34(s,3H).

Sixth step: tert-butyl (3 ar,6s,6 as) -6- ((5-chloro-2, 4-difluorophenyl) (methyl) aminocarbonyl) -3 a-cyclopropyl-2, 2-dimethyltetrahydro-5H- [1,3] dioxolo [4,5-c ] pyrrole-5-carboxylic acid ester (B11-7)

tert-butyl(3aR,6S,6aS)-6-((5-chloro-2,4-difluorophenyl)(methyl)carbamoyl)-3a-cyclopropyl-2,2-dimethyltetrahydro-5H-[1,3]dioxolo[4,5-c]pyrrole-5-carboxylate

Tert-butyl (3 aR,6S,6 aS) -6- ((5-chloro-2, 4-difluorophenyl) aminocarbonyl) -3 a-cyclopropyl-2, 2-dimethyltetrahydro-5H- [1,3] dioxolo [4,5-c ] pyrrole-5-carboxylate (200 mg, 422. Mu. Mol) and cesium carbonate (275 mg, 845. Mu. Mol) were dissolved in N, N-dimethylformamide (1.00 mL), and after stirring the reaction solution for 30 minutes, methyl iodide (120 mg, 845. Mu. Mol) was slowly added dropwise, and the reaction solution was stirred at 25℃for 2 hours. After completion of the reaction, water (10 mL) was added to dilute the reaction, followed by extraction with ethyl acetate (10 mL x 3), and the organic phase was washed with saturated sodium chloride solution (10 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give tert-butyl (3 ar,6s,6 as) -6- ((5-chloro-2, 4-difluorophenyl) (methyl) aminocarbonyl) -3 a-cyclopropyl-2, 2-dimethyltetrahydro-5H- [1,3] dioxolo [4,5-c ] pyrrole-5-carboxylate (B11-7, 180mg, crude) which was used directly in the next step.

LC-MS,M/Z(ESI):387.1[M+H-100] ⁺

Seventh step: tert-butyl (3 aS,6S,6 aS) -6- ((5-chloro-2, 4-difluorophenyl) (methyl) aminocarbonyl) -3 a-cyclopropyl-2, 2-dimethyl-4-oxolanyl tetrahydro-5H- [1,3] dioxolo [4,5-c ] pyrrole-5-carboxylic acid ester (B11-8)

tert-butyl(3aS,6S,6aS)-6-((5-chloro-2,4-difluorophenyl)(methyl)carbamoyl)-3a-cyclopropyl-2,2-dimethyl-4-oxotetrahydro-5H-[1,3]dioxolo[4,5-c]pyrrole-5-carboxylate

Ruthenium dioxide hydrate (13.6 mg, 90.3. Mu. Mol) and sodium periodate (289.91 mg,1.36 mmol) were dissolved in water (1.50 mL) under nitrogen atmosphere, nitrogen was replaced three times, the reaction solution was stirred at 25℃for 30 minutes, and then tert-butyl (3 aR,6S,6 aS) -6- ((5-chloro-2, 4-difluorophenyl) (methyl) aminocarbonyl) -3 a-cyclopropyl-2, 2-dimethyltetrahydro-5H- [1,3] dioxolo [4,5-c ] pyrrole-5-carboxylate (B11-7, 220mg, 451.81. Mu. Mol) dissolved in ethyl acetate (1.00 mL) was slowly added to the reaction solution, which was stirred at 25℃for 12 hours. After completion of the reaction, the reaction was quenched by addition of saturated sodium sulfite (20.0 mL), then extracted with ethyl acetate (10 mL x 3), and the organic phase was washed with saturated sodium chloride solution (20 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate=5:1 to 1:1 as mobile phase) to give tert-butyl (3 as,6s,6 as) -6- ((5-chloro-2, 4-difluorophenyl) (methyl) aminocarbonyl) -3 a-cyclopropyl-2, 2-dimethyl-4-oxotetrahydro-5H- [1,3] dioxolo [4,5-c ] pyrrole-5-carboxylate (B11-8, 105mg, 46.4% yield).

Eighth step: (3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -6 a-cyclopropyl-N, 2-trimethyl-6-oxolanyl tetrahydro-4H- [1,3] dioxolo [4,5-c ] pyrrole-4-carboxamide (B11-9)

(3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-6a-cyclopropyl-N,2,2-trimethyl-6-oxotetrahydro-4H-[1,3]dioxolo[4,5-c]pyrrole-4-carboxamide

Tert-butyl (3 aS,6S,6 aS) -6- ((5-chloro-2, 4-difluorophenyl) (methyl) aminocarbonyl) -3 a-cyclopropyl-2, 2-dimethyl-4-oxotetrahydro-5H- [1,3] dioxolo [4,5-c ] pyrrole-5-carboxylate (B11-8, 50.0mg, 99.8. Mu. Mol) was dissolved in dichloromethane (1.00 mL), and then zinc bromide (89.9 mg, 399. Mu. Mol) was added and the reaction mixture was stirred at 25℃for 12 hours. After completion of the reaction, the reaction was quenched with saturated sodium bicarbonate (10 mL) and then extracted with dichloromethane (10 mL x 3) and the organic phase was washed with saturated sodium chloride solution (10 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give the crude, (3 as,4s,6 as) -N- (5-chloro-2, 4-difluorophenyl) -6 a-cyclopropyl-N, 2-trimethyl-6-oxotetrahydro-4H- [1,3] dioxolo [4,5-c ] pyrrole-4-carboxamide (B11-9, 35.0mg, 87.5% yield).

Ninth step: (3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -6 a-cyclopropyl-N, 2-trimethyl-5- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -6-oxotetrahydro-4H- [1,3] dioxolo [4,5-c ] pyrrole-4-carboxamide (B11-10)

(3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-6a-cyclopropyl-N,2,2-trimethyl-5-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-6-oxotetrahydro-4H-[1,3]dioxolo[4,5-c]pyrrole-4-carboxamide

(3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -6 a-cyclopropyl-N, 2-trimethyl-6-oxolanyl tetrahydro-4H- [1,3] dioxolo [4,5-c ] pyrrole-4-carboxamide (B11-9, 50.0mg, 124. Mu. Mol) and 2-chloro-6-methyl-4- (trifluoromethyl) pyridine (36.6 mg,18. Mu. Mol) were dissolved in 1, 4-dioxane (2.00 mL), then potassium carbonate (51.7 mg, 374. Mu. Mol), tris (dibenzylideneacetone) dipalladium (10.7 mg, 18.7. Mu. Mol) and 4, 5-bis (diphenylphosphorus) -9, 9-dimethylxanthene (21.6 mg, 37.4. Mu. Mol) were added, respectively, and after three nitrogen substitutions, the reaction solution was stirred at 100℃for 3 hours. After the completion of the reaction, the crude product was concentrated by direct filtration to give a brown oily substance, which was isolated and purified by column chromatography (petroleum ether: ethyl acetate=3:1 as mobile phase) to give (3 as,4s,6 as) -N- (5-chloro-2, 4-difluorophenyl) -6 a-cyclopropyl-N, 2-trimethyl-5- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -6-oxotetrahydro-4H- [1,3] dioxolo [4,5-c ] pyrrole-4-carboxamide (B11-10, 60.0mg, yield 85.9%).

LC-MS,M/Z(ESI):560.0[M+H] ⁺

Seventeenth step: (2S, 3S, 4S) -N- (5-chloro-2, 4-difluorophenyl) -4-cyclopropyl-3, 4-dihydroxy-N-methyl-1- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -5-oxoy-lidene pyrrolidine-2-carboxamide (I-11).

((2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-4-cyclopropyl-3,4-dihydroxy-N-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-carboxamide

(3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -6 a-cyclopropyl-N, 2-trimethyl-5- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) scheme6-Oxyltetrahydro-4H- [1,3]Dioxapenta [4,5-c]Pyrrole-4-carboxamide (B11-10, 20mg, 35.72. Mu. Mol) was dissolved in dichloromethane (1.00 mL), then a solution of boron trichloride in dichloromethane (1M, 71.4. Mu.L) was added at-20℃and the reaction solution was stirred at 25℃for 2 hours. After the reaction was completed, ph=9 was adjusted with saturated sodium bicarbonate solution, then extracted with dichloromethane (10 mL x 3), and the organic phase was washed with saturated sodium chloride solution (10.0 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product is separated and purified by reverse phase high performance liquid chromatography, and the separation method comprises the following steps: waters Xbridge C18 is 150 x 50mm x 10um; mobile phase [ water (NH) ₄ HCO ₃ ) -acetonitrile]The method comprises the steps of carrying out a first treatment on the surface of the Gradient 50% -80% B over 10min, purification gives (2S, 3S, 4S) -N- (5-chloro-2, 4-difluorophenyl) -4 cyclopropyl-3, 4-dihydroxy-N-methyl-1- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -5-oxopyrrolidine-2-carboxamide (I-11, 12.0mg, 20.9% yield).

¹ H NMR(400MHz,CH ₃ OH-d ₄ )δ8.32-8.52(m,1H),7.95-8.13(m,1H),7.45–7.54(m,1H),7.22–7.34(m,1H),5.08–5.92(m,1H),4.55-4.60(m,1H),3.96-4.02(m,1H),3.24–3.68(m,1H),2.52-2.70(m,1H),0.87-1.22(m,1H),0.28-0.60(m,4H).

EXAMPLE 6 preparation of Compound I-20

The synthetic route is as follows:

the first step: 2-chloro-4-cyclopropyl-6-methylpyridine (B20-2)

2-chloro-4-cyclopropyl-6-methylpyridine

2-chloro-4-iodo-6-methylpyridine (501 mg,1.98 mmol) and cyclopropylboronic acid (220.72 mg,2.57 mmol) were dissolved in 1, 4-dioxane (5.00 mL), then potassium phosphate (1.26 g,5.93 mmol), palladium acetate (66.56 mg, 296.49. Mu. Mol) and tricyclohexylphosphine (166.29 mg, 592.98. Mu. Mol) were added respectively under nitrogen atmosphere, and after three nitrogen substitutions, the reaction solution was stirred at 100℃for 3 hours. After the reaction was completed, the crude product was concentrated by direct filtration to give a brown oily substance, which was isolated and purified by column chromatography (petroleum ether: ethyl acetate=15:1 as mobile phase) to give 2-chloro-4-cyclopropyl-6-methylpyridine (150 mg, yield 45.3%).

¹ H NMR(400MHz,CHCl ₃ -d)δ＝6.79(s,1H),6.73(s,1H),2.47(s,3H),1.79-1.83(m,1H),1.04-1.13(m,2H),0.73-0.83(m,2H).

And a second step of: (3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -5- (4-cyclopropyl-6-methylpyridin-2-yl) -2, 6 a-trimethyl-N- (methyl) -6-oxotetrahydro-4H- [1,3] dioxolo [4,5-c ] pyrrole-4-carboxamide (B20-3)

(3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-5-(4-cyclopropyl-6-methylpyridin-2-yl)-2,2,6a-trimethyl-N-(methyl)-6-oxotetrahydro-4H-[1,3]dioxolo[4,5-c]pyrrole-4-carboxamide

(3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -2, 6 a-trimethyl-N- (methyl) -6-oxotetrahydro-4H- [1,3] dioxolo [4,5-c ] pyrrole-4-carboxamide (B5-10, 150mg, 400.25. Mu. Mol) and 2-chloro-4-cyclopropyl-6-methylpyridine (134.19 mg, 800.50. Mu. Mol) were dissolved in 1, 4-dioxane (2.00 mL), then cesium carbonate (260.82 mg, 800.50. Mu. Mol), tris (dibenzylideneacetone) dipalladium (54.98 mg, 60.04. Mu. Mol) and 4, 5-bis (diphenylphosphorus) -9, 9-dimethylxanthene (69.48 mg, 120.07. Mu. Mol) were added, respectively, and after three nitrogen substitutions, the reaction solution was stirred at 100℃for 3 hours. After the reaction, the crude product of brown oily matter is obtained by direct filtration and concentration, and the crude product is separated and purified by column chromatography (petroleum ether: ethyl acetate=3:1 as mobile phase), and (3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -5- (4-cyclopropyl-6-methylpyridin-2-yl) -2, 6 a-trimethyl-N- (methyl) -6-oxotetrahydro-4H- [1,3] dioxolo [4,5-c ] pyrrole-4-carboxamide (B20-3, 20.0mg, crude product) is obtained after purification.

LC-MS,M/Z(ESI):506.1[M+H] ⁺

And a third step of: (2S, 3S, 4S) -N- (5-chloro-2, 4-difluorophenyl) -1- (4-cyclopropyl-6-methylpyridin-2-yl) -3, 4-dihydroxy-4-methyl-N- (methyl) -5-oxopyrrolidine-2-carboxamide (I-20)

(2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-1-(4-cyclopropyl-6-methylpyridin-2-yl)-3,4-dihydroxy-4-methyl-N-(methyl-d3)-5-oxopyrrolidine-2-carboxamide

3aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -5- (4-cyclopropyl-6-methylpyridin-2-yl) -2, 6 a-trimethyl-N- (methyl) -6-oxotetrahydro-4H- [1,3]Dioxapenta [4,5-c]Pyrrole-4-carboxamide (B20-3, 10mg, 19.77. Mu. Mol) was dissolved in dichloromethane (1.00 mL), then a solution of boron trichloride in dichloromethane (1M, 39.5L) was added at-20℃and the reaction was stirred at 25℃for 1 hour. After the reaction was completed, ph=9 was adjusted with saturated sodium bicarbonate solution, then extracted with dichloromethane (10 mL x 2), and the organic phase was washed with saturated sodium chloride solution (10.0 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product is separated and purified by reverse phase high performance liquid chromatography, and the separation method comprises the following steps: waters Xbridge 150 x 25mm x 5um; mobile phase [ water (NH) ₄ HCO ₃ ) -acetonitrile]The method comprises the steps of carrying out a first treatment on the surface of the Gradient 36% -66% Bover 9min, after purification, (2S, 3S, 4S) -N- (5-chloro-2, 4-difluorophenyl) -1- (4-cyclopropyl-6-methylpyridin-2-yl) -3, 4-dihydroxy-4-methyl-N- (methyl-d 3) -5-oxopyrrolidine-2-carboxamide (I-20, 0.75mg, 3.64% yield).

¹ H NMR(400MHz,CH ₃ OH-d ₄ )δ＝8.09(t,1H),7.74-7.85(m,1H),7.30-7.39(m,1H),7.18(t,1H),3.97-4.03(m,1H),3.15(s,2H),2.26-2.43(m,3H),1.93(d,1H),1.76-1.82(m,1H),1.31-1.37(m,1H),0.94-1.00(m,2H),0.67-0.73(m,1H).

EXAMPLE 7 preparation of Compound I-21

The synthetic route is as follows:

the first step: methyl 4- (trifluoromethyl) picolinic acid ester

Methyl 4-(trifluoromethyl)picolinate

2-bromo-4- (trifluoromethyl) pyridine (30.0 g,132 mmol) was dissolved in methanol (300 mL), triethylamine (26.8 g,265 mmol) and [1, 1-bis (diphenylphosphorus) ferrocene ] palladium dichloride (1.94 g,2.65 mmol) were added under nitrogen atmosphere, the reaction solution was replaced three times with carbon monoxide gas, and then stirred under carbon monoxide (50.0 Psi) gas pressure at 80℃for 12 hours. After completion of the reaction, the mixture was directly filtered and concentrated under reduced pressure to give a brown liquid, which was then diluted with water (500 mL), extracted with ethyl acetate (300 ml×3), and the organic phase was washed with saturated sodium chloride solution (500 mL) and dried over anhydrous sodium sulfate, and then concentrated by filtration to give methyl 4- (trifluoromethyl) picolinate (27.2 g, crude product).

LCMS,M/Z(ESI):206.1[M+H] ⁺

And a second step of: (4- (trifluoromethyl) pyridin-2-yl) methanol

(4-(trifluoromethyl)pyridin-2-yl)methanol

Methyl 4- (trifluoromethyl) methylpyridine ester (10.0 g,48.7 mmol) was dissolved in ethanol (100 mL) and then sodium borohydride (3.69 g,97.5 mmol) was added in portions under nitrogen at 0℃and the reaction stirred at 25℃for 12 hours. After completion of the reaction, quench by slow addition of saturated aqueous ammonium chloride (50 mL) at 0 ℃ and extraction with ethyl acetate (100 mL x 3), the organic phase was washed with saturated aqueous sodium chloride (200 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give (4- (trifluoromethyl) pyridin-2-yl) methanol (4.00 g, crude) which was used directly in the next step.

¹ H NMR(400MHz,CHCl ₃ -d)δ8.63(d,1H),7.54(s,1H),7.34(d,1H),4.78(s,2H).

And a third step of: 4- (trifluoromethyl) picoline aldehyde

4-(trifluoromethyl)picolinaldehyde

(4- (trifluoromethyl) pyridin-2-yl) methanol (4.00 g,22.6 mmol) was dissolved in tetrahydrofuran (40.0 mL), and manganese dioxide (19.6 g,225 mmol) was then added in portions and the reaction stirred at 70℃for 1 hour. After the completion of the reaction, the reaction solution was directly filtered and concentrated under reduced pressure to give 4- (trifluoromethyl) picolinaldehyde (1.40 g, crude product), which was directly used in the next step.

¹ H NMR(400MHz,CHCl ₃ -d)δ10.14(s,1H),9.00(d,1H),8.18(s,1H),7.64-7.88(m,1H),7.76(dd,0.94Hz,1H).

Fourth step: 2- (difluoromethyl) -4- (trifluoromethyl) pyridine

2-(difluoromethyl)-4-(trifluoromethyl)pyridine

4- (trifluoromethyl) picolinaldehyde (1.50 g,18.5 mmol) was dissolved in dichloromethane (10.0 mL) and then bis (2-methoxyethyl) aminothiotrifluoride (3.75 g,37.1 mmol) was added at 0deg.C. The reaction solution was stirred at 25℃for 2 hours. After completion of the reaction, the reaction was quenched by addition of saturated sodium bicarbonate solution at 0deg.C until no more bubbles were generated, then extracted with dichloromethane (10 mL), and the organic phases were combined to give 2- (difluoromethyl) -4- (trifluoromethyl) pyridine (1.00 g, crude product) which was used directly in the next step.

Fifth step: 2- (difluoromethyl) -4- (trifluoromethyl) pyridine 1-oxide

2-(difluoromethyl)-4-(trifluoromethyl)pyridine 1-oxide

2- (difluoromethyl) -4- (trifluoromethyl) pyridine (2.00 g,10.1 mmol) was dissolved in dichloromethane (20.0 mL), then trifluoroacetic acid (5.78 g,50.7 mmol) and hydrogen peroxide (71.0 mmol,6.82mL,30% purity) were added at 0deg.C, and the reaction was stirred at 25deg.C for 12 hours. After completion of the reaction, the reaction was quenched by addition of saturated sodium sulfite (10 mL), then extracted with dichloromethane (10 mL x 3), and the organic phase was washed with saturated sodium chloride solution (20 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate=5:1-3:1) to give 2- (difluoromethyl) -4- (trifluoromethyl) pyridine 1-oxide (600 mg, yield 27.8%).

¹ H NMR(400MHz,CHCl ₃ -d)δ8.33(d,1H),7.86(d,1H),7.62(dd,2.06Hz,1H),7.22(t,1H).

Sixth step: 2-chloro-6- (difluoromethyl) -4- (trifluoromethyl) pyridine (B21-1)

2-chloro-6-(difluoromethyl)-4-(trifluoromethyl)pyridine

2- (difluoromethyl) -4- (trifluoromethyl) pyridine 1-oxide (160 mg, 505. Mu. Mol) was dissolved in phosphorus oxychloride (3.60 g,23.4 mmol), and the reaction solution was stirred at 100℃for 3 hours. After completion of the reaction, the reaction mixture was slowly quenched by addition to water (10.0 mL), then washed with saturated sodium bicarbonate, extracted with ethyl acetate (10.0 mL x 2), and the organic phase was washed with saturated sodium chloride solution (10 mL) and dried over anhydrous sodium sulfate, filtered and concentrated at 30 ℃ to give 2-chloro-6- (difluoromethyl) -4- (trifluoromethyl) pyridine (B21-1) (150 mg, crude product) which was used directly in the next step.

Seventh step: tert-butyl (3 ar,6s,6 as) -6- ((5-chloro-2, 4-difluorophenyl) (methyl-d 3) aminocarbonyl) -2, 3 a-trimethyltetrahydro-5H- [1,3] dioxolo [4,5-c ] pyrrole-5-carboxylic acid ester (B21-2)

tert-butyl(3aR,6S,6aS)-6-((5-chloro-2,4-difluorophenyl)(methyl-d3)carbamoyl)-2,2,3a-trimethyltetrahydro-5H-[1,3]dioxolo[4,5-c]pyrrole-5-carboxylate

Tert-butyl (3 aR,6S,6 aS) -6- ((5-chloro-2, 4-difluorophenyl) aminocarbonyl) -2, 3 a-trimethyltetrahydro-5H- [1,3] dioxolo [4,5-c ] pyrrole-5-carboxylic acid ester (B5-7) (3.20 g,7.16 mmol) and cesium carbonate (4.67 g,14.32 mmol) were dissolved in N, N-dimethylformamide (30.0 mL), and after stirring the reaction solution at room temperature for 30 minutes, deuterated iodomethane (3.56 g,25.06 mmol) was slowly added dropwise and the reaction solution was stirred at 25℃for 2 hours. After completion of the reaction, water (200 mL) was added to dilute the reaction, followed by extraction with ethyl acetate (30 mL x 3) and the organic phase was washed with saturated sodium chloride solution (60 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give tert-butyl (3 ar,6s,6 as) -6- ((5-chloro-2, 4-difluorophenyl) (methyl-d 3) aminocarbonyl) -2, 3 a-trimethyltetrahydro-5H- [1,3] dioxolo [4,5-c ] pyrrole-5-carboxylate (B21-2) (2.00 g, crude) which was used directly in the next step.

LC-MS,M/Z(ESI):364.1[M+H] ⁺

Eighth step: tert-butyl (3 aS,6S,6 aS) -6- ((5-chloro-2, 4-difluorophenyl) (methyl-d 3) aminocarbonyl) -2, 3 a-trimethyl-4-oxotetrahydro-5H- [1,3] dioxolo [4,5-c ] pyrrole-5-carboxylic acid ester (B21-3)

tert-butyl(3aS,6S,6aS)-6-((5-chloro-2,4-difluorophenyl)(methyl-d3)carbamoyl)-2,2,3a-trimethyl-4-oxotetrahydro-5H-[1,3]dioxolo[4,5-c]pyrrole-5-carboxylate

Ruthenium trichloride hydrate (29 mg,1.29 mmol) and sodium periodate (2.77 g,12.9 mmol) were dissolved in water (30.0 mL) under nitrogen atmosphere, the reaction solution was stirred at 25 ℃ for 30 minutes by nitrogen substitution three times, then tert-butyl (3 ar,6s,6 as) -6- ((5-chloro-2, 4-difluorophenyl) (methyl-d 3) aminocarbonyl) -2, 3 a-trimethyltetrahydro-5H- [1,3] dioxolo [4,5-c ] pyrrole-5-carboxylate (B21-2) (2.00 g,4.31 mmol) dissolved in ethyl acetate (30.0 mL) was slowly added to the reaction solution, and finally the reaction solution was stirred at 25 ℃ for 12 hours. After completion of the reaction, the reaction was quenched by addition of saturated sodium sulfite (30.0 mL), then extracted with ethyl acetate (30 mL x 3), and the organic phase was washed with saturated sodium chloride solution (20 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate=5:1 to 1:1 as mobile phase) to give tert-butyl (3 as,6s,6 as) -6- ((5-chloro-2, 4-difluorophenyl) (methyl-d 3) aminocarbonyl) -2, 3 a-trimethyl-4-oxotetrahydro-5H- [1,3] dioxolo [4,5-c ] pyrrole-5-carboxylate (B21-3) (1.00 g, 48.5% yield).

LC-MS,M/Z(ESI):378.1[M+H] ⁺

Ninth step: (3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -2, 6 a-trimethyl-N- (methyl-d 3) -6-oxotetrahydro-4H- [1,3] dioxolo [4,5-c ] pyrrole-4-carboxamide (B21-4)

(3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-2,2,6a-trimethyl-N-(methyl-d3)-6-oxotetrahydro-4H-[1,3]dioxolo[4,5-c]pyrrole-4-carboxamide

Tert-butyl (3 aS,6S,6 aS) -6- ((5-chloro-2, 4-difluorophenyl) (methyl-d 3) aminocarbonyl) -2, 3 a-trimethyl-4-oxotetrahydro-5H- [1,3] dioxolo [4,5-c ] pyrrole-5-carboxylate (B21-3) (1.00 g,2.09 mmol) was dissolved in dichloromethane (10.0 mL), and then zinc bromide (2.36 g,10.4 mmol) was added and the reaction stirred at 25℃for 12 hours. After completion of the reaction, the reaction was quenched with saturated sodium bicarbonate (100 mL) and then extracted with dichloromethane (50 mL x 3), the organic phase was washed with saturated sodium chloride solution (50 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give (3 as,4s,6 as) -N- (5-chloro-2, 4-difluorophenyl) -2, 6 a-trimethyl-N- (methyl-d 3) -6-oxotetrahydro-4H- [1,3] dioxolo [4,5-c ] pyrrole-4-carboxamide (B21-4) (500 mg, crude product, 63.2%)

¹ H NMR(400MHz,CHCl ₃ -d)δ7.61–7.67(m,1H),7.08–7.27(m,1H),6.09(d,1H),4.01(d,1H),3.43(d,1H),1.35-1.38(m,9H).

Tenth step: (3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -5- (6- (difluoromethyl) -4- (trifluoromethyl) pyridin-2-yl) -2, 6 a-trimethyl-N- (methyl-d 3) -6-oxotetrahydro-4H- [1,3] dioxolo [4,5-c ] pyrrole-4-carboxamide (B21-5)

(3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-5-(6-(difluoromethyl)-4-(trifluoromethyl)pyridin-2-yl)-2,2,6a-trimethyl-N-(methyl-d3)-6-oxotetrahydro-4H-[1,3]dioxolo[4,5-c]pyrrole-4-carboxamide

(3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -2, 6 a-trimethyl-N- (methyl-d 3) -6-oxybutylene tetrahydro-4H- [1,3] dioxolo [4,5-c ] pyrrole-4-carboxamide (B21-4) (180 mg,544 umol) and 2-chloro-6-methyl-4- (trifluoromethyl) pyridine (B21-1) (122.58 mg, 529.40. Mu. Mol) were dissolved in 1, 4-dioxane (2.00 mL), and cesium carbonate (172.49 mg, 529.40. Mu. Mol), tris (dibenzylideneacetone) dipalladium (36.36 mg, 39.71. Mu. Mol) and 4, 5-bis (diphenylphosphorus) -9, 9-dimethylxanthene (45.95 mg, 79.41. Mu.) were added, respectively, and after three nitrogen substitutions, the reaction solution was stirred at 100℃for 3 hours. After completion of the reaction, crude product of brown oil was obtained by direct filtration and concentration, and the crude product was isolated and purified by prep-TLC (petroleum ether: ethyl acetate=3:1) to give (3 as,4s,6 as) -N- (5-chloro-2, 4-difluorophenyl) -5- (6- (difluoromethyl) -4- (trifluoromethyl) pyridin-2-yl) -2, 6 a-trimethyl-N- (methyl-d 3) -6-oxotetrahydro-4H- [1,3] dioxolo [4,5-c ] pyrrole-4-carboxamide (B21-5) (60.0 mg, yield 37.5%).

LC-MS,M/Z(ESI):573.1[M+H] ⁺

Eleventh step: (2S, 3S, 4S) -N- (5-chloro-2, 4-difluorophenyl) -1- (6- (difluoromethyl) -4- (trifluoromethyl) pyridin-2-yl) -3, 4-dihydroxy-4-methyl-N- (methyl-d 3) -5-oxopyrrolidine-2-carboxamide (I-21)

(2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-1-(6-(difluoromethyl)-4-(trifluoromethyl)pyridin-2-yl)-3,4-dihydroxy-4-methyl-N-(methyl-d3)-5-oxopyrrolidine-2-carboxamide

(3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -5- (6- (difluoromethyl) -4- (trifluoromethyl) pyridin-2-yl) -2, 6 a-trimethyl-N- (methyl-d 3) -6-oxotetrahydro-4H- [1,3]Dioxapenta [4,5-c]Pyrrole-4-carboxamide (B21-5) (60.0 mg, 104. Mu. Mol) was dissolved in dichloromethane (1.00 mL), then a solution of boron trichloride in dichloromethane (1M, 314. Mu.L) was added at-20℃and the reaction was stirred at 25℃for 2 hours. After the reaction was completed, ph=9 was adjusted with saturated sodium bicarbonate solution, then extracted with dichloromethane (10 mL x 3), and the organic phase was washed with saturated sodium chloride solution (20.0 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product is separated and purified by reverse phase high performance liquid chromatography, and the separation method comprises the following steps: waters Xbridge 150 x 25mm x 5um; mobile phase [ water (NH) ₄ HCO ₃ ) -acetonitrile]The method comprises the steps of carrying out a first treatment on the surface of the 28% -58%, for 8min, to give (2S, 3S, 4S) -N- (5-chloro-2, 4-difluorophenyl) -1- (6- (difluoromethyl) -4- (trifluoromethyl) pyridin-2-yl) -3, 4-dihydroxy-4-methyl-N- (methyl-d 3) -5-oxopyrrolidine-2-carboxamide (I-21) (8.65 mg, 16.5% yield).

¹ H NMR(400MHz,CH ₃ OH-d ₄ )δ8.60-8.95(m,1H),8.10-8.23(m,1H),7.63(s,1H),6.60-7.27(m,1H),6.38-6.85(m,1H),5.25–5.75(m,1H),4.49(s,1H)，4.13–4.48(m,1H),3.05–3.26(m,1H),1.42–1.64(m,1H).

EXAMPLE 8 preparation of Compound I-22

The synthetic route is shown below:

the first step: synthesis of (3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -2, 6 a-trimethyl-N- (methyl-d 3) -5- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -6-carbonyltetrahydro-4H- [1,3] dioxazolo [4,5-c ] pyrrole-4-carboxamide (B22-1)

(3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-2,2,6a-trimethyl-N-(methyl-d3)-5-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-6-oxotetrahydro-4H-[1,3]dioxolo[4,5-c]pyrrole-4-carboxamide

(3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -2, 6 a-trimethyl-N- (methyl-d 3) -6-carbonyltetrahydro-4H- [1,3] dioxazolo [4,5-c ] pyrrole-4-carboxamide (B21-4) (5.6 g,14.8 mmol), potassium carbonate (6.2 g,44.4 mmol), tris (dibenzylideneacetone) dipalladium (1.36 g,1.48 mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (900 mg,1.48 mmol), 2-chloro-6-methyl-4- (trifluoromethyl) pyridine (3.5 g,17.76 mmol) were dissolved in a solution of 1, 4-dioxane (80 mL), the reaction system was replaced with argon three times, and then reacted at 100℃for 12H. The reaction solution was filtered, the filtrate was washed with water, dried, and concentrated to give a crude product, which was purified by silica gel column (ethyl acetate/petroleum ether=1/5) to give (3 as,4s,6 as) -N- (5-chloro-2, 4-difluorophenyl) -2, 6 a-trimethyl-N- (methyl-d 3) -5- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -6-carbonyltetrahydro-4H- [1,3] dioxazolo [4,5-c ] pyrrole-4-carboxamide (B22-1) (5.1 g, yield 63.7%).

LC-MS,M/Z(ESI):537.1[M+H] ⁺

And a second step of: (2S, 3S, 4S) -N- (5-chloro-2, 4-difluorophenyl) -3, 4-dihydroxy-4-methyl-N- (methyl-d 3) -1- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -5-carbonylpyrrolidine-2-carboxamide (I-22)

(2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-3,4-dihydroxy-4-methyl-N-(methyl-d3)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-carboxamide

Under the protection of nitrogen, the (3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -2, 6 a-trimethyl-N- (methyl-d 3) -5- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -6-oxo-tetrahydro-4H- [1,3 ]Dioxazolo [4,5-c ]]Pyrrole-4-carboxamide (B22-1) (5.1 g,9.5 mmol) was dissolved in dichloromethane (50 mL) and BCl was added at 0deg.C ₃ Is a solution in methylene chloride (19 mL,19mmol,1 mol/L). The reaction mixture was stirred for 2h at 0deg.C, quenched with saturated sodium bicarbonate solution and adjusted to pH=8-9 after completion, extracted with a mixed solvent of (dichloromethane/methanol=20/1, 30mL×2) and the combined organic phases dried over anhydrous sodium sulfate, concentrated to give crude product, which was purified by HPLC to give (2S, 3S, 4S) -N- (5-chloro-2, 4-difluorophenyl) -3, 4-dihydroxy-4-methyl-N- (methyl-d 3) -1- (6-methyl-4- (trifluoromethyl) pyridin-2-yl) -5-carbonylpyrrolidine-2-carboxamide (I-22) (2.7 g, 57.4% yield).

¹ H NMR(400MHz,CH ₃ OH-d4)δ8.57–8.39(m,1H),8.19–8.06(m,1H),7.53–7.25(m,2H),5.87–5.20(m,1H),4.50–3.94(m,1H),2.72-2.50(m,3H),1.45–1.26(m,3H).

EXAMPLE 9 preparation of Compound I-23

The synthetic route is as follows:

the first step: 6-chloro-3-fluoro-4-iodo-2-methylpyridine

6-chloro-3-fluoro-4-iodo-2-methylpyridine

6-chloro-3-fluoro-2-methyl-pyridine (3.00 g,20.6 mmol) was dissolved in tetrahydrofuran (30.0 mL), lithium diisopropylamide (2.5M, 10.7 mL) was added under nitrogen atmosphere at-78℃and the reaction solution was stirred at-78℃for 30 minutes, then elemental iodine (10.5 g,41.2 mmol) dissolved in tetrahydrofuran (20.0 mL) was slowly added dropwise to the reaction solution, and finally the reaction solution was stirred at-78℃for 1.5 hours. After the completion of the reaction, the reaction mixture was slowly added dropwise to a saturated ammonium chloride solution at 0 ℃, then extracted with ethyl acetate (20 mL. Times.3), and the organic phase was washed with a saturated sodium chloride solution (50 mL) and dried over anhydrous sodium sulfate, and then concentrated by filtration to give a crude product as a yellow solid, which was isolated and purified by column chromatography (mobile phase: petroleum ether: ethyl acetate=100:1-5:1) to give 6-chloro-3-fluoro-4-iodo-2-methylpyridine (3.00 g, yield 53.6%).

¹ H NMR(400MHz,DMSO-d ₆ )δ7.85(d,1H)2.41(d,3H)

And a second step of: 6-chloro-3-fluoro-2-methyl-4- (trifluoromethyl) pyridine

6-chloro-3-fluoro-2-methyl-4-(trifluoromethyl)pyridine

6-chloro-3-fluoro-4-iodo-2-methylpyridine (4.00 g,14.7 mmol) was dissolved in N, N-dimethylformamide (80.0 mL), and then cuprous iodide (8.42 g,44.2 mmol) and methyl 2, 2-difluoro-2-fluorosulfonyl-acetate (11.3 g,58.9 mmol) were added at 25℃and the reaction stirred at 100℃for 3 hours. After completion of the reaction, diluted with water (100 mL) and extracted with ethyl acetate (200 mL x 3), the organic phase was washed with saturated sodium chloride solution (60.0 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give 6-chloro-3-fluoro-2-methyl-4- (trifluoromethyl) pyridine (2.50 g, crude) which was used directly in the next step. LC-MS, M/Z (ESI): 214.1[ M+H ]] ⁺

And a third step of: (3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -5- [ 5-fluoro-6-methyl-4- (trifluoromethyl) pyridin-2-yl]-N-( ² H ₃ ) Methyl-2, 6 a-trimethyl-6-oxy-hexahydro- [1,3 ]]Dioxapenta [4,5-c]Pyrrole-4-carboxamide (B23-2)

(3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-5-[5-fluoro-6-methyl-4-(trifluoromethyl)pyridin-2-yl]-N-( ² H ₃ )methyl-2,2,6a-trimethyl-6-oxo-hexahydro-[1,3]dioxolo[4,5-c]pyrrole-4-carboxamide

(3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -2, 6 a-trimethyl-N- (methyl-d 3) -6-oxotetrahydro-4H- [1,3]Dioxapenta [4,5-c]Pyrrole-4-carboxamide (B21-4) (300 mg, 794. Mu. Mmol) and 6-chloro-3-fluoro-2-methyl-4- (trifluoromethyl) pyridine (B23-1) (508 mg,2.38 mmol) were dissolved in 1, 4-dioxane (3.00 mL), cesium carbonate (517mg, 1.59 mmol), tris (dibenzylideneacetone) dipalladium (109 mg, 119. Mu. Mol) and 4, 5-bis (diphenylphosphorus) -9, 9-dimethylxanthene (109 mg, 119. Mu. Mol) were then added under nitrogen protection, respectively, and after three substitutions of nitrogen, the reaction solution was stirred at 90℃for 12 hours. After the reaction is finished, directly filtering and concentrating to obtain a brown oily crude product, separating and purifying the crude product by prep-TLC (petroleum ether: ethyl acetate=3:1 as mobile phase), and separating and purifying the crude product by reverse phase high performance liquid chromatography, wherein the analytical method is as follows: watersXbridge 150 x 25mm x 5um; mobile phase [ water (NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the Gradient 60% -90% B over 9min, obtaining (3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -5- (6- (difluoromethyl) -4- (trifluoromethyl) pyridin-2-yl) -2, 6 a-trimethyl-N- (methyl-d 3) -6-oxotetrahydro-4H- [1,3]Dioxapenta [4,5-c]Pyrrole-4-carboxamide (B23-2) (80.0 mg, 18.6% yield). LC-MS, M/Z (ESI): 555.0[ M+H ]] ⁺

Fourth step: (2S, 3S, 4S) -N- (5-chloro-2, 4-difluorophenyl) -1- [ 5-fluoro-6-methyl-4- (trifluoromethyl) pyridin-2-yl]-3, 4-dihydroxy-N- ² H ₃ ) Methyl-4-methyl-5-oxopyrrolidine-2-carboxamide (I-23)

(2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-1-[5-fluoro-6-methyl-4-(trifluoromethyl)pyridin-2-yl]-3,4-dihydroxy-N-( ² H ₃ )methyl-4-methyl-5-oxopyrrolidine-2-carboxamide

(3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -5- (6- (difluoro)Methyl) -4- (trifluoromethyl) pyridin-2-yl) -2, 6 a-trimethyl-N- (methyl-d 3) -6-oxotetrahydro-4H- [1,3]Dioxapenta [4,5-c]Pyrrole-4-carboxamide (B23-2) (60.0 mg, 108. Mu. Mol) was dissolved in dichloromethane (1.00 mL), then a solution of boron trichloride in dichloromethane (1M, 324. Mu.L) was added at-20℃and the reaction was stirred at 25℃for 2 hours. After the reaction was completed, the pH was adjusted to 9 with saturated sodium bicarbonate solution, then extracted with dichloromethane (10 mL x 3), and the organic phase was washed with saturated sodium chloride solution (20.0 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product is separated and purified by reverse phase high performance liquid chromatography, and the separation method comprises the following steps: waters Xbridge 150 x 25mm x 5um; mobile phase: [ Water (NH) ₄ HCO ₃ ) -acetonitrile]The method comprises the steps of carrying out a first treatment on the surface of the B percent is 28 to 58 percent, and the purification is carried out for 8min to obtain (2S, 3S, 4S) -N- (5-chloro-2, 4-difluorophenyl) -1- [ 5-fluoro-6-methyl-4- (trifluoromethyl) pyridin-2-yl]-3, 4-dihydroxy-N- ² H ₃ ) Methyl-4-methyl-5-oxopyrrolidine-2-carboxamide (I-23) (43.0 mg, 77.2% yield).

¹ H NMR(400MHz,CH ₃ OH-d ₄ )δ8.45-8.69(m,1H)8.04-8.18(m,1H)7.43-7.66&7.30-7.34(m,1H)5.10-5.97(m,1H)3.97-4.55(m,1H)2.51-2.71(m,3H)1.30-1.49(m,3H)。

EXAMPLE 10 preparation of Compound I-24

The synthetic route is as follows:

the first step: 2-bromo-6-/v ² H ₃ ) Methyl-4- (trifluoromethyl) pyridine

2-bromo-6-( ² H ₃ )methyl-4-(trifluoromethyl)pyridine

2, 6-dibromo-4- (trifluoromethyl) pyridine (2.00 g,6.56 mmol) was dissolved in tetrahydrofuran (60.0 mL) and N-methylpyrrolidone (4.00 mL) and acetyl was added under nitrogen atmosphereIron acetonate (231.67 mg, 655.97. Mu. Mol) and magnesium deuterated methane iodide (1M, 11.81 mL) were stirred at 25℃for 1 hour. After the reaction is completed, the mixture is quenched by saturated ammonium chloride (30.0 mL), extracted by ethyl acetate (300 mL. Times.3), washed by saturated sodium chloride solution (500 mL) and dried by anhydrous sodium sulfate, filtered and concentrated to obtain 2-bromo-6- ² H ₃ ) Methyl-4- (trifluoromethyl) pyridine (B24-1) (27.2 g, crude).

LCMS,M/Z(ESI):244.9[M+H] ⁺

And a second step of: (3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -N- (-, A-N-alpha-hydroxy-ethyl-phenyl) ² H ₃ ) Methyl-2, 6 a-trimethyl-5- [6- ] alpha ] ² H ₃ ) Methyl-4- (trifluoromethyl) pyridin-2-yl]-6-oxo-hexahydro- [1,3 ]]Dioxapenta [4,5-c ]Pyrrole-4-carboxamide (B24-2)

(3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-N-( ² H ₃ )methyl-2,2,6a-trimethyl-5-[6-( ² H ₃ )methyl-4-(trifluoromethyl)pyridin-2-yl]-6-oxo-hexahydro-[1,3]dioxolo[4,5-c]pyrrole-4-carboxamide

(3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -2, 6 a-trimethyl-N- (methyl-d 3) -6-oxotetrahydro-4H- [1,3]Dioxapenta [4,5-c]Pyrrole-4-carboxamide (B21-4) (300 mg, 794. Mu. Mol) and 2-chloro-6-methyl-4- (trifluoromethyl) pyridine (578 mg,2.38 mmol) were dissolved in 1, 4-dioxane (3.00 mL), then cesium carbonate (300 mg, 794. Mu. Mol), tris (dibenzylideneacetone) dipalladium (109 mg, 119. Mu. Mol) and 4, 5-bis (diphenylphosphorus) -9, 9-dimethylxanthene (137 mg, 238. Mu. Mol) were added respectively under nitrogen atmosphere, and after three nitrogen substitutions, the reaction solution was stirred at 100℃for 12 hours. After the reaction is finished, directly filtering and concentrating to obtain a brown oily crude product, separating and purifying the crude product by column chromatography (the mobile phase is petroleum ether: ethyl acetate=3:1), and obtaining (3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -N- & lt- & gt after purification ² H ₃ ) Methyl-2, 6 a-trimethyl-5- [6- ] alpha ] ² H ₃ ) Methyl-4- (trifluoromethyl) pyridin-2-yl]-6-oxygenSubunit-hexahydro- [1,3 ]]Dioxapenta [4,5-c]Pyrrole-4-carboxamide (B24-2) (60.0 mg, 14.0% yield).

LC-MS,M/Z(ESI):540.3[M+H] ⁺

And a third step of: (2S, 3S, 4S) -N- (5-chloro-2, 4-difluorophenyl) -3, 4-dihydroxy-N- ² H ₃ ) Methyl-4-methyl-1- [6- ], process for preparing same and use thereof ² H ₃ ) Methyl-4- (trifluoromethyl) pyridin-2-yl ]-5-Oxypyrrolidine-2-carboxamide (I-24)

(2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-3,4-dihydroxy-N-( ² H ₃ )methyl-4-methyl-1-[6-( ² H ₃ )methyl-4-(trifluoromethyl)pyridin-2-yl]-5-oxopyrrolidine-2-carboxamide

(3 aS,4S,6 aS) -N- (5-chloro-2, 4-difluorophenyl) -N- ² H ₃ ) Methyl-2, 6 a-trimethyl-5- [6- ] alpha ] ² H ₃ ) Methyl-4- (trifluoromethyl) pyridin-2-yl]-6-oxo-hexahydro- [1,3 ]]Dioxapenta [4,5-c]Pyrrole-4-carboxamide (B24-2) (60.0 mg, 111. Mu. Mol) was dissolved in dichloromethane (1.00 mL), then a dichloromethane solution of boron trichloride (1M, 333. Mu.L) was added at-20℃and the reaction solution was stirred at 25℃for 2 hours. After the reaction was completed, the pH was adjusted to 9 with saturated sodium bicarbonate solution, then extracted with dichloromethane (10 mL x 3), and the organic phase was washed with saturated sodium chloride solution (20.0 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product is separated and purified by reverse phase high performance liquid chromatography, and the separation method comprises the following steps: waters Xbridge 150 x 25mm x 5um; mobile phase [ water (NH) ₄ HCO ₃ ) -acetonitrile]The method comprises the steps of carrying out a first treatment on the surface of the B percent is 28 to 58 percent, and the purification is carried out for 8min to obtain (2S, 3S, 4S) -N- (5-chloro-2, 4-difluorophenyl) -3, 4-dihydroxy-N- ² H ₃ ) Methyl-4-methyl-1- [6- ], process for preparing same and use thereof ² H ₃ ) Methyl-4- (trifluoromethyl) pyridin-2-yl]-5-Oxopyrrolidine-2-carboxamide (I-24) (54.0 mg, 97.2% yield).

¹ H NMR(400MHz,CH ₃ OH-d ₄ )δppm 8.49-8.54(m,1H)8.11-8.26(m,1H)7.46-7.58(m,1H)7.30-7.49(m,1H)5.25-5.91(m,1H)4.00-4.55(m,1H)1.31-1.50(m,3H).

The following compounds can be prepared by reference to the preparation methods of examples 1-10:

effect test example 1

In the test examples of the present invention, the preparation method of the control compound I is referred to in patent WO2021/028643A1, the preparation method of the control compound II is referred to in patent WO2021028670A1, and the structures are shown below.

Control Compound I

Control Compound II

Test example 1 Compounds inhibition test of Pol theta polymerase Activity

The recombinant Pol θ polymerase domain (aa 1819-2590) used in this test was expressed and purified by the Invitrogen Bac-to-Bac baculovirus expression system. The assay used to measure Pol theta polymerase activity was a PicoGreen dsDNA quantitative assay.

The reaction system consisted of compounds dissolved in DMSO, purified recombinant Pol theta (aa 1819-2590) protein, annealing mixtures and dNTPs. Wherein all compounds were prepared in 10mM stock solution in DMSO, and in assay buffer (25mM Tris HCl pH 7.5, 12.5mM NaCl,0.5mM MgCl) ₂ 5% glycerol, 0.01% Triton X-100,0.01% BGG and 1mM DTT); the recombinant protein solution is obtained by dissolving recombinant Pol theta (aa 1819-2590) protein with the final concentration of 30nM in an experiment buffer solution; the annealing mixture is obtained by dissolving a primer (5'-GCG GCT GTC ATA AG-3') with the final concentration of 150nM and a template (5 ' -GCT ACA TTG ACA ATG GCA TCA AAT CTC AGA TTG CGT CTT ATG ACA GCC) with the final concentration of 150nM in an experiment buffer solution, heating to 42 ℃ for 3min and annealing; the dNTP solution was obtained by dissolving dNTP having a final concentration of 120. Mu.M in the assay buffer.

In the compound screening experiments, 2. Mu.L/well of recombinant protein solution was added to 384 well plates (Perkin Elmer-Proxiplate), followed by 0.06. Mu.L/well of different concentrations of compound and negative control (DMSO) in sequence, and finally 2. Mu.L/well of annealing mix and 2. Mu.L/well of dNTPs. The blank contains only annealing mixture and dntps. After the 384-well plate was sealed with an aluminum foil sealing film, it was incubated at 37℃for 30min, then the reaction was stopped by adding 4. Mu.L of a reaction stop solution (25 mM Tris-HCl,10mM cationic EDTA and 1:80 PicoGreen) to the test well, and incubated at room temperature for 90min under light-shielding conditions, and then incubated in an ELISA (Perkinelmer2104. λex=485 nm λem=520 nm). Inhibition was calculated by calculating the IC of each compound using GraphPad Prism8 software, inhibition = (1- (compound group-blank)/(negative control group-blank)) ×100% ₅₀ Values. The results are shown in Table 1.

TABLE 1 inhibition of Pol theta polymerase Activity by Compounds

Test compounds	IC ₅₀ (nM)
		Control Compound I	30
Control Compound II	86
		I-1A	16
I-1B	14
		I-4	35
I-5	5
		I-21	73
I-22	22
		I-23	21
I-24	45

The experimental results show that: the compound has good inhibition effect on Pol theta polymerase.

Test example 2 Compounds inhibit tumor cell proliferation assay

DLD-1BRCA2 (-/-) cells were seeded at a density of 500 cells/100. Mu.L/well onto 96 well cell plates and the cell plates were placed in an incubator for 18 hours (37 ℃,5% CO) ₂ ). The next day of drug treatment was started, 100. Mu.L/well of gradient diluted test compound solution (initial concentration of each drug was 100. Mu.M, dilution was DMSO, dilution ratio was 1:3, nine gradient points for each drug) or DMSO (negative control) was added to the culture plate medium, a blank group was additionally provided without inoculating cell lines to which only the medium was added, and after the addition of the drugs, the culture plate was placed in an incubator for further incubation for 5 days (37 ℃ C., 5% CO) ₂ ). On the fourth and seventh days, replacement of fresh medium and compounds was performed, respectively. Detection was performed on the tenth day, 100. Mu.L of CellTiter-Glo (Promega G9243) reagent was added to each well, and after shaking for 5 minutes, the mixture was allowed to stand at room temperature for 5 minutes, using an enzyme-labeled instrument (Perkinelmer)2104 Chemiluminescent signal values were determined for each well. Cell proliferation inhibition = (1- (compound group-blank)/(DMSO group-blank)) ×100%, IC of each compound was calculated using GraphPad Prism8 software ₅₀ Values. The results are shown in Table 2.

TABLE 2 inhibition of tumor cell proliferation by Compounds

Test compounds	IC ₅₀ (μM)
		Control Compound II	15
I-5	3.31
		I-21	5.54
I-22	4.96

Experimental results indicate that the compounds of the invention show an inhibitory effect on DLD-1 BRCA2 (-/-) cell proliferation.

Test example 3: pharmacokinetic test in mice

Male ICR mice, 20-25g, were used, fasted overnight. 3 mice were taken, and the dosage of oral gavage administration was 10mg/kg, and the administration volume was 10mL/kg. In addition, 3 mice were taken, and the intravenous administration dose was 3mg/kg and the administration volume was 5mL/kg. Blood was collected 5, 15, 30 minutes and 1, 2, 4, 6, 8, 24 hours before and after dosing. Blood samples 6800g, centrifuged at 6000g at 2-8deg.C for 3 min, and plasma was collected by separation and stored at-80deg.C. Plasma at each time point is taken, 3-5 times of acetonitrile solution containing an internal standard is added for mixing, vortex mixing is carried out for 1 minute, 13000 r/min, centrifugation is carried out at 4 ℃ for 10 minutes, 3 times of water is added for mixing the supernatant, and a proper amount of mixed solution is taken for LC-MS/MS analysis. Pharmacokinetic parameters were calculated using Phoenix WinNonlin8.2.0 by blood concentration data at different time points to provide parameters such as CL, vz, AUC0-T, AUC 0-infinity, MRT 0-infinity, cmax, tmax, and T1/2, as well as mean and standard deviation thereof. The results are shown in tables 3 and 4.

TABLE 3 results of the test compounds in a mouse pharmacokinetic assay

TABLE 4 results of the test compounds in a mouse pharmacokinetic assay

Experimental results show that the compound of the invention has high oral exposure, good pharmacokinetic property and good patentability.

While embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and not to be construed as limiting the invention, and that variations, modifications, alternatives and variations may be made to the above embodiments by one of ordinary skill in the art within the scope of the invention.

Claims

1. A heterocyclic compound shown in a formula I, a stereoisomer or a pharmaceutically acceptable salt thereof,

wherein m is 0, 1, 2, 3 or 4;

n is 0, 1, 2, 3 or 4;

ring K is a 5-6 membered heteroaromatic ring;

R ₃ wherein each R is _a Each independently is deuterium, halogen, hydroxy, amino, cyano, C ₁ -C ₆ Alkyl or C ₁ -C ₆ A haloalkyl group;

or, R ₄ And R is ₅ Each independently is R _b ，

Or, R ₆ And R is ₇ Each independently is R _b ，

each R is _b Each of said R _a Each independently is deuterium, halogen, hydroxy, amino, cyano, C ₁ -C ₆ Alkyl or C ₁ -C ₆ A haloalkyl group;

2. The heterocyclic compound of formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as claimed in claim 1, wherein the heterocyclic compound of formula I satisfies one or more of the following conditions (1) to (36):

(1) In the ring K, in the 5-6 membered heteroaromatic ring, a heteroatom is selected from one or more of N, O and S, and the number of the heteroatom is 1, 2 or 3; for example, in ring K, the 5-6 membered heteroaromatic ring is a 6 membered heteroaromatic ring, preferably, the 5-6 membered heteroaromatic ring has a heteroatom selected from N, more preferably, the 5-6 membered heteroaromatic ring has 1 heteroatom;

(2)R ₁ and R is ₂ Wherein each halogen is independently fluorine, chlorine, bromine or iodine, such as chlorine or fluorine;

(3)R ₁ and R is ₂ In said formula (I), said R being unsubstituted or substituted by one or more R _a Substituted C ₁ -C ₆ C in alkyl ₁ -C ₆ Alkyl is each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl;

(4)R ₁ and R is ₂ In said formula (I), said R being unsubstituted or substituted by one or more R _a Substituted C ₁ -C ₆ C in alkoxy ₁ -C ₆ Alkoxy groups are each independently methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy or tert-butoxy;

(5)R ₁ and R is ₂ Wherein each of the 3-6 membered cycloalkyl groups is independently cyclopropane, cyclobutane, cyclopentane or cyclohexane, for example cyclopropane;

(6)R ₁ And R is ₂ Wherein each R is _a Wherein each halogen is independently fluorine, chlorine, bromine or iodine, such as fluorine;

(7)R ₁ and R is ₂ Wherein each R is _a In the above, the C ₁ -C ₆ Alkyl is each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl;

(8)R ₁ and R is ₂ Wherein each R is _a In the above, the C ₁ -C ₆ Haloalkyl is each independently C1, 2 or 3 halogen substituents ₁ -C ₆ An alkyl group;

(9)R ₃ in said formula (I), said R being unsubstituted or substituted by one or more R _a Substituted C ₁ -C ₆ C in alkyl ₁ -C ₆ Alkyl is each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl;

(10)R ₃ wherein each R is _a Wherein each halogen is independently fluorine, chlorine, bromine or iodine, such as fluorine;

(11)R ₃ wherein each R is _a In the above, the C ₁ -C ₆ Alkyl is each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl;

(12)R ₃ wherein each R is _a In the above, the C ₁ -C ₆ Haloalkyl is each independently C1, 2 or 3 halogen substituents ₁ -C ₆ An alkyl group;

(13)R ₄ 、R ₅ 、R ₆ 、R ₇ and R is ₈ In the above, the C ₁ -C ₆ Alkyl is each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl;

(14) The R is _b Wherein each halogen is independently fluorine, chlorine, bromine or iodine, such as fluorine;

(15) Each R is _b In said, unsubstituted or R _a Substituted C ₁ -C ₆ C in alkyl ₁ -C ₆ Alkyl is each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl;

(16) Each R is _b In said, unsubstituted or R _a Substituted C ₁ -C ₆ C in alkoxy ₁ -C ₆ Alkoxy groups are each independently methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy or tert-butoxy;

(17) Each R is _b In said, unsubstituted or R _a The 3-6 membered cycloalkyl groups in the substituted 3-6 membered cycloalkyl groups are each independently cyclopropane, cyclobutane, cyclopentane or cyclohexane groups, for example cyclopropane groups;

(18) Each R is _b In said, unsubstituted or R _a In the 4-6 membered heterocycloalkyl group in the substituted 4-6 membered heterocycloalkyl group, a heteroatom is selected from one or more of N, O and S, and the number of the heteroatom is 1, 2 or 3;

(19) Each R is _b Wherein each R is _a Wherein each halogen is independently fluorine, chlorine, bromine or iodine, such as fluorine;

(20) Each R is _b Wherein each R is _a In the above, the C ₁ -C ₆ Alkyl is each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl;

(21) Each R is _b Wherein each R is _a In the above, the C ₁ -C ₆ Haloalkyl is each independently C1, 2 or 3 halogen substituents ₁ -C ₆ An alkyl group;

(22) In each ring A, said R is unsubstituted or substituted by one or more _a The 3-8 membered cycloalkyl group in the substituted 3-8 membered cycloalkyl group is a 3-6 membered cycloalkyl group, for example, the 3-8 membered cycloalkyl groups are each independently a cyclopropanyl group, a cyclobutyl group, a cyclopentanyl group or a cyclohexenyl group, preferably a cyclopropanyl group;

(23) In each ring A, the radicals are unsubstituted or substituted by R _a The 4-8 membered heterocycloalkyl group in the substituted 4-8 membered heterocycloalkyl group is a 4-6 membered heterocycloalkyl group, preferably the 4-6 membered heterocylic groupIn the cycloalkyl, the heteroatom is selected from one or more of N, O and S, and the heteroatom number is 1, 2 or 3;

(24) In each ring A, each R _a Wherein each halogen is independently fluorine, chlorine, bromine or iodine;

(25) In each ring A, each R _a In the above, the C ₁ -C ₆ Alkyl is each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl;

(26) In each ring A, each R _a In the above, the C ₁ -C ₆ Haloalkyl is each independently C1, 2 or 3 halogen substituents ₁ -C ₆ An alkyl group;

(27) When m is 2, 3, 4, said R ₁ The same or different;

(28) When n is 2, 3 or 4, R is ₂ The same or different;

(29) When R is _a When there are a plurality of R _a The same or different;

(30) The "or" is merely "or", and a relationship is selected between the expression elements;

(31) When the alkyl group is unsubstituted, the alkyl group is unsubstituted;

(32) When the alkyl group is not illustrated as isotopically substituted, the alkyl group is one that does not contain carbon and/or hydrogen isotopes;

(33) When the cycloalkyl or carbocyclyl is not described as an unsaturated or partially saturated carbocycle, the cycloalkyl or carbocyclyl is a saturated cycloalkyl or carbocyclyl;

(34) When the heterocycloalkyl group is not described as an unsaturated or partially saturated heterocycloalkyl group, the heterocycloalkyl group is a saturated heterocycloalkyl group;

(35) When the heteroaryl ring radical is not illustrated as a heteroaryl ring radical fused to a benzene ring, the heteroaryl ring radical is an unfused heteroaryl ring radical;

(36) The stereoisomers are enantiomers or non-corresponding isomers.

3. The heterocyclic compound of formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as claimed in claim 1, wherein the heterocyclic compound of formula I is any one of the following schemes:

scheme one: in the heterocyclic compound shown in the formula I,

wherein m and n are integers of 0 to 4 respectively;

R ₃ is hydrogen, C ₁ -C ₆ Alkyl or C ₁ -C ₆ A haloalkyl group;

or R is ₇ And R is ₈ And to which they are attachedThe C atoms bound together form a ring A, R ₄ 、R ₅ 、R ₆ Each independently is hydrogen or C ₁ -C ₆ An alkyl group;

the ring A is 3-8 membered cycloalkyl or 4-8 membered heterocycloalkyl;

the ring a is optionally substituted with a substituent selected from the group consisting of: halogen, hydroxy, amino, cyano, C ₁ -C ₆ Alkyl, C ₁ -C ₆ A haloalkyl group;

scheme II: in the heterocyclic compound shown in the formula I,

wherein m is 0, 1, 2, 3, 4;

n is 0, 1, 2, 3, 4;

R ₁ 、R ₂ each independently selected from: halogen, hydroxy, amino, cyano, C unsubstituted or substituted by one or more Ra ₁ -C ₆ Alkyl, C unsubstituted or substituted by one or more Ra ₁ -C ₆ Alkoxy, 3-6 membered cycloalkyl, unsubstituted or substituted with one or more Ra;

When m is 2, 3, 4, said R ₁ The same or different;

when n is 2, 3 or 4, R is ₂ The same or different;

R ₃ c being hydrogen, unsubstituted or substituted by one or more Ra ₁ -C ₆ An alkyl group;

R ₄ 、R ₅ 、R ₆ 、R ₇ each independently is hydrogen or Rb, R ₈ Is Rb;

or R is ₄ 、R ₅ 、R ₆ 、R ₇ 、R ₈ Each independently is hydrogen or Rb, and R ₄ 、R ₅ 、R ₆ 、R ₇ 、R ₈ At least one of which is cyano;

or R is ₄ And R is ₅ Rb, or R, which are identical or different ₄ And R is ₅ With the C-atom to which they are attachedTogether with the sub-groups forming a ring A, R ₆ 、R ₇ 、R ₈ Each independently is hydrogen or Rb;

or R is ₅ And R is ₆ Together with the C atom to which they are attached form a ring A, R ₄ 、R ₇ 、R ₈ Each independently is hydrogen or Rb;

or R is ₆ And R is ₇ Rb, or R, which are identical or different ₆ And R is ₇ Together with the C atom to which they are attached form a ring A, R ₄ 、R ₅ 、R ₈ Each independently is hydrogen or Rb;

or R is ₇ And R is ₈ Together with the C atom to which they are attached form a ring A, R ₄ 、R ₅ 、R ₆ Each independently is hydrogen or Rb;

wherein each Rb is independently halogen, hydroxy, amino, cyano, C unsubstituted or substituted with Ra ₁ -C ₆ Alkyl, C unsubstituted or substituted by Ra ₁ -C ₆ An alkoxy group;

any of the above Ra are each independently halogen, hydroxy, amino, cyano, C ₁ -C ₆ Alkyl, C ₁ -C ₆ A haloalkyl group;

when Ra is plural, the Ra is the same or different;

scheme III: in the heterocyclic compound shown in the formula I,

wherein m is 0, 1, 2, 3, 4;

n is 0, 1, 2, 3, 4;

ring K is a 5-6 membered heteroaromatic ring;

when m is 2, 3, 4, said R ₁ The same or different;

when n is 2, 3 or 4, R is ₂ The same or different;

or R is ₄ And R is ₅ Rb, or R, which are identical or different ₄ And R is ₅ Together with the C atom to which they are attached form a ring A, R ₆ 、R ₇ 、R ₈ Each independently is hydrogen or Rb;

wherein each Rb is independently halogen, hydroxy, amino, cyano, C unsubstituted or substituted with Ra ₁ -C ₆ Alkyl, C unsubstituted or substituted by Ra ₁ -C ₆ Alkoxy, 3-6 membered cycloalkyl unsubstituted or substituted by Ra, 4-6 membered heterocycle unsubstituted or substituted by RaAn alkyl group;

when Ra is plural, the Ra is the same or different;

scheme IV: in the heterocyclic compound shown in the formula I,

wherein m is 0, 1, 2, 3, 4;

n is 0, 1, 2, 3, 4;

ring K is a 5-6 membered heteroaromatic ring;

when m is 2, 3, 4, said R ₁ The same or different;

When n is 2, 3 or 4, R is ₂ The same or different;

or R is ₄ And R is ₅ Is Rb which may be the same or different,or R is ₄ And R is ₅ Together with the C atom to which they are attached form a ring A, R ₆ 、R ₇ 、R ₈ Each independently is hydrogen or Rb;

wherein each Rb is independently halogen, hydroxy, amino, cyano, C unsubstituted or substituted with Ra ₁ -C ₆ Alkyl, C unsubstituted or substituted by Ra ₁ -C ₆ Alkoxy, 3-6 membered cycloalkyl unsubstituted or substituted by Ra, 4-6 membered heterocycloalkyl unsubstituted or substituted by Ra;

when Ra is plural, the Ra is the same or different.

4. The heterocyclic compound of formula I, a stereoisomer thereof or a pharmaceutically acceptable salt thereof as described in claim 3, wherein the heterocyclic compound of formula I is any one of the following schemes:

scheme one: the heterocyclic compound shown in the formula I has a structure II

Wherein m and n are integers of 0 to 4 respectively;

the ring A is 3-8 membered cycloalkyl or 4-8 membered heterocycloalkyl;

the ring A is optionally selectedSubstituted from the following substituents: halogen, hydroxy, amino, cyano, C ₁ -C ₆ Alkyl, C ₁ -C ₆ A haloalkyl group;

scheme II: the heterocyclic compound shown in the formula I has a structure II

Wherein m and n are integers of 0 to 4 respectively;

R ₃ is hydrogen, C ₁ -C ₆ Alkyl or C ₁ -C ₆ A haloalkyl group;

the ring A is 3-8 membered cycloalkyl or 4-8 membered heterocycloalkyl;

5. The heterocyclic compound of the formula I, a stereoisomer thereof or a pharmaceutically acceptable salt thereof according to claim 3 or 4, wherein the heterocyclic compound of the formula I has the structure Ia or Ib

Wherein m, n, R ₁ 、R ₂ 、R ₃ 、R ₄ 、R ₅ 、R ₆ 、R ₇ And R is ₈ As claimed in claim 3 or 4.

6. The heterocyclic compound of the formula I, a stereoisomer thereof or a pharmaceutically acceptable salt thereof as described in claim 4,having the structure->R ₁₁ Is C ₁ -C ₃ Alkyl, R ₁₂ Is C ₁ -C ₃ A haloalkyl group;

preferably, the method comprises the steps of,having the structure->

7. The heterocyclic compound of the formula I, a stereoisomer thereof or a pharmaceutically acceptable salt thereof as described in claim 4,having the structure->R ₂₁ 、R ₂₂ 、R ₂₃ Halogen which is the same or different; preferably, said R ₂₁ 、R ₂₂ 、R ₂₃ Each independently F or Cl;

more preferably, the process is carried out,having the structure->

8. The heterocyclic compound of the formula I, a stereoisomer thereof or a pharmaceutically acceptable salt thereof as described in claim 3 or 4, wherein R ₄ And R is ₅ Together with the C atom to which they are attached form a ring A, R ₆ 、R ₇ 、R ₈ Each independently is hydrogen, hydroxy, C ₁ -C ₆ Alkyl, C ₁ -C ₆ A haloalkyl group;

or R is ₆ And R is ₇ Together with the C atom to which they are attached form a ring A, R ₄ 、R ₅ 、R ₈ Each independently is hydrogenHydroxy, C ₁ -C ₆ Alkyl, C ₁ -C ₆ A haloalkyl group;

or R is ₇ And R is ₈ Together with the C atom to which they are attached form a ring A, R ₄ 、R ₅ 、R ₆ Each independently is hydrogen, hydroxy, C ₁ -C ₆ Alkyl, C ₁ -C ₆ A haloalkyl group;

preferably, R ₄ And R is ₅ Together with the C atom to which they are attached form a ring A, R ₆ 、R ₇ 、R ₈ Each independently is hydrogen;

the ring A is 3-6 membered cycloalkyl or 4-8 membered heterocycloalkyl;

9. The heterocyclic compound of the formula I, a stereoisomer thereof or a pharmaceutically acceptable salt thereof as described in claim 3 or 4, wherein R ₄ 、R ₅ 、R ₆ 、R ₇ Is hydrogen, R ₈ Is C ₁ -C ₃ An alkyl group.

10. The heterocyclic compound of the formula I, a stereoisomer thereof or a pharmaceutically acceptable salt thereof as described in claim 3 or 4, wherein R ₄ And R is ₅ R being identical or different _b ，R ₆ 、R ₇ 、R ₈ Each independently is hydrogen, hydroxy, halogen, C ₁ -C ₆ Alkyl, C ₁ -C ₆ A haloalkyl group;

R _b is halogen, hydroxy, cyano, C ₁ -C ₆ An alkyl group;

preferably, R _b Fluorine, chlorine, hydroxyl, cyano, methyl, ethyl, propyl.

11. The heterocyclic compound of formula I, a stereoisomer thereof or a pharmaceutically acceptable salt thereof according to claim 3, wherein the heterocyclic compound of formula I has the structure Ic, id or Ie

Wherein the ring K, m, n, R ₁ 、R ₂ 、R ₃ 、R ₄ 、R ₅ 、R ₆ 、R ₇ And R is ₈ A method as claimed in claim 3.

12. The heterocyclic compound of formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as described in claim 3 or 11, wherein ring K is a 5-or 6-membered N-containing heteroaryl ring; preferably, ring K is a pyridine ring, pyrimidine ring, pyrazine ring, pyridazine ring.

13. The heterocyclic compound of the formula I as described in any one of claims 3 to 4 and claim 8, a stereochemistry thereofAn isomer or a pharmaceutically acceptable salt thereof, characterized in that R ₃ Is deuterated methyl.

14. The heterocyclic compound of formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to claim 1 or 2, wherein the heterocyclic compound of formula I satisfies one or more of the following conditions (1) to (21):

(1) M is 2 or 3, for example 2;

(2) Said n is 1 or 3, e.g. 3,

(3) The ring K is a 5-6 membered heteroaromatic ring, wherein in the 5-6 membered heteroaromatic ring, a heteroatom is N, and the number of heteroatoms is 1 or 2, for example, the ring K is pyridine;

(4) Each R is ₁ Each independently is halogen, unsubstituted or substituted with one or more R _a Substituted C ₁ -C ₆ Alkyl or 3-6 membered cycloalkyl, preferably each R ₁ Each independently is halogen, unsubstituted or substituted with one or more R _a Substituted C ₁ -C ₆ Alkyl, e.g. each R ₁ Each independently is halogen, trifluoromethyl, methyl or tridentate methyl, as well as, for example, each R ₁ Each independently is unsubstituted or substituted with one or more R _a Substituted C ₁ -C ₆ An alkyl group;

(5) When m is 2, each R ₁ Ortho or para to the heteroatom in ring K;

(6) When m is 3, each R ₁ Adjacent and in the ortho, meta or para position to the heteroatom in ring K;

(7) Each R is ₁ Wherein each R is _a Each independently is halogen or deuterium, e.g., each R ₁ Wherein each R is _a Each independently is halogen;

(8)R ₂ each independently is halogen;

(9)R ₃ unsubstituted or substituted by one or more R _a Substituted C ₁ -C ₆ Alkyl radicals, e.g. R ₃ Is C ₁ -C ₆ Alkyl, e.g. R ₃ To be covered by one or more R _a Substituted C ₁ -C ₆ Alkyl group；

(10)R ₃ Wherein each R is _a Each independently deuterium;

(11) Each R is _b Each independently is halogen, hydroxy, cyano, 3-6 membered cycloalkyl or is unsubstituted or substituted with one or more R _a Substituted C ₁ -C ₆ Alkyl, preferably each R _b Each independently is halogen, hydroxy or C ₁ -C ₆ Alkyl, e.g. each R _b Each independently is hydroxy or C ₁ -C ₆ An alkyl group;

(12) Each R is _b Wherein each R is _a Each independently is halogen;

(13) Each ring A is independently 3-8 membered cycloalkyl;

(14)R ₄ 、R ₅ 、R ₆ and R is ₇ Each independently is hydrogen or R _b ，R ₈ Is C ₁ -C ₆ An alkyl group;

(15)R ₅ 、R ₆ 、R ₇ and R is ₈ Each independently is hydrogen or R _b And R is ₄ Is cyano;

(16)R ₄ 、R ₅ 、R ₇ and R is ₈ Each independently is hydrogen or R _b And R is ₆ Is cyano;

(17)R ₄ and R is ₅ Each independently is R _b ，R ₆ 、R ₇ And R is ₈ Each independently is R _b Or hydrogen, or a combination of hydrogen and,

Preferably, R ₄ And R is ₅ Each independently is R _b ，R ₆ And R is ₇ Each independently is hydrogen or hydroxy, R ₈ Is hydrogen;

(18)R ₆ and R is ₇ Each independently is R _b ，R ₄ 、R ₅ And R is ₈ Each independently is R _b Or hydrogen, or a combination of hydrogen and,

for example, R ₆ And R is ₇ Each independently is R _b ，R ₄ And R is ₅ Each independently is hydrogen, C ₁ -C ₆ Alkyl or hydroxy, R ₈ Is hydrogen;

(19)R ₄ and R is ₅ Together with the C atom to which they are attached form a ring A, R ₆ And R is ₇ Each independently is hydrogen or hydroxy, R ₈ Is hydrogen, e.g. R ₄ And R is ₅ Together with the C atom to which they are attached form a ring A, R ₆ And R is ₇ Each independently is hydrogen, R ₈ Is hydrogen;

(20)R ₅ and R is ₆ Together with the C atom to which they are attached form a ring A, R ₄ And R is ₇ Each independently is hydrogen or hydroxy, R ₈ Is hydrogen;

(21)R ₆ and R is ₇ Together with the C atom to which they are attached form a ring A, R ₄ And R is ₅ Each independently is hydrogen or hydroxy, R ₈ Is hydrogen.

15. The heterocyclic compound of formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as claimed in claim 1, wherein the heterocyclic compound of formula I is any one of the following schemes:

scheme one:

m is 0, 1, 2, 3 or 4;

n is 0, 1, 2, 3 or 4;

each R is ₁ Wherein each R is _a Each independently is halogen or deuterium;

R ₂ each independently is halogen;

R ₃ wherein each R is _a Each independently deuterium;

each R is _b Wherein each R is _a Each independently is halogen;

each ring A is independently 3-8 membered cycloalkyl;

scheme II:

m is 2 or 3;

n is 3;

ring K is pyridine;

each R is ₁ Wherein each R is _a Each independently is halogen or deuterium;

R ₂ each independently is halogen;

R ₃ wherein each R is _a Each independently deuterium;

each ring A is independently 3-8 membered cycloalkyl;

scheme III:

m is 2 or 3;

n is 3;

ring K is pyridine;

R ₂ Each independently is halogen;

R ₃ wherein each R is _a Each independently deuterium;

each R is _b Each independently is hydroxy or C ₁ -C ₆ An alkyl group;

each ring A is independently 3-8 membered cycloalkyl;

scheme IV:

m is 2;

n is 3;

ring K is pyridine;

when m is 2, each R ₁ Ortho or para to the heteroatom in ring K;

each R is ₁ Wherein each R is _a Each independently is halogen;

R ₂ each independently is halogen;

R ₃ is C ₁ -C ₆ An alkyl group;

each R is _b Each independently is hydroxy or C ₁ -C ₆ An alkyl group;

each ring A is independently 3-8 membered cycloalkyl;

scheme five:

m is 2 or 3;

n is 3;

ring K is pyridine;

each R is ₁ Wherein each R is _a Each independently is halogen or deuterium;

R ₂ each independently is halogen;

R ₃ wherein each R is _a Each independently deuterium;

each R is _b Each independently is hydroxy or C ₁ -C ₆ An alkyl group;

each ring a is independently a 3-8 membered cycloalkyl.

16. The heterocyclic compound of formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as claimed in claim 1, wherein the heterocyclic compound of formula I satisfies one or more of the following conditions (1) to (11):

(1) Ring K is pyridine or pyrimidine;

(2)R ₁ methyl, trifluoromethyl, difluoromethyl, cyclopropanyl, fluoro or tridentate methyl;

(3)R ₂ is fluorine or chlorine;

(4)R ₃ methyl or tridentate methyl;

(5)R ₄ and R is ₅ Each independently is hydrogen, fluoro, hydroxy, methyl, cyclopropane, trifluoromethylCyano or difluoromethyl;

alternatively, R ₄ And R is ₅ Together with the C atom to which they are attached, form a ring a, which is cyclopropenyl:

Alternatively, R ₅ And R is ₆ Together with the C atom to which they are attached, form a ring a, which is cyclopropane group;

(6)R ₆ and R is ₇ Each independently is hydrogen, hydroxy, methyl or cyano;

(7)R ₈ is hydrogen or methyl;

(8)is->

(9)Is->

(10)Is->

(11)Is->

17. The heterocyclic compound of formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as claimed in claim 1, wherein the heterocyclic compound of formula I is any one of the following compounds:

18. the heterocyclic compound of formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as claimed in claim 1, wherein the heterocyclic compound of formula I is any one of the following compounds:

19. a pharmaceutical composition comprising a heterocyclic compound of formula I according to any one of claims 1 to 18, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and optionally

i) One or more active agents; and/or

ii) a pharmaceutically acceptable carrier.

20. Use of a heterocyclic compound of formula I according to any one of claims 1 to 18, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 19, comprising:

1) Inhibiting Pol theta activity;

2) Preventing and treating Pol theta mediated diseases;

preferably, the Pol θ -mediated disease is a cancer, for example the cancer is lymphoma, rhabdoid tumor, multiple myeloma, uterine cancer, gastric cancer, peripheral nervous system cancer, rhabdomyosarcoma, bone cancer, colorectal cancer, mesothelioma, breast cancer, ovarian cancer, lung cancer, fibroblast cancer, central nervous system cancer, urinary tract cancer, upper respiratory tract cancer, leukemia, kidney cancer, skin cancer, esophageal cancer and pancreatic cancer.