TW202341987A - Pol[Theta] inhibitor - Google Patents

Abstract

Provided is a Pol[Theta] inhibitor. Specifically provided are a heterocyclic compound represented by formula I, and a stereoisomer or pharmaceutically acceptable salt thereof, wherein m and n respectively are integers from 0 to 4; and a ring K is a 5-6 membered heteroaryl ring. The heterocyclic compound has a good inhibitory effect on a Pol[Theta] polymerase, and can prevent or treat diseases or conditions mediated by Pol[Theta].

Description

A Polθ inhibitor

This application claims priority to Chinese patent application 2022102395700 with a filing date of 2022/3/11. This application claims the priority of Chinese patent application 2022105905480 with a filing date of 2022/5/26. This application claims the priority of Chinese patent application 2022106895452 with a filing date of 2022/6/16. This application claims the priority of Chinese patent application 2022109820918 with a filing date of 2022/8/16. This application claims the priority of Chinese patent application 2023102038322 with a filing date of 2023/3/03. This application cites the full text of the above-mentioned Chinese patent application.

The invention belongs to the field of medicine. Specifically, the invention relates to a Polθ inhibitor.

Synthetic lethality is an emerging research direction in the field of anti-tumor. Among them, treatment targeting DNA repair pathways is the focus of this patent. Some tumor cells will lose a certain DNA repair pathway due to genetic mutations, making them overly dependent on the remaining DNA repair pathways. At this time, targeted inhibition of this remaining DNA repair pathway can specifically inhibit the DNA repair pathway. Kills such tumor cells but has no killing effect on normal body cells. In recent years, poly(ADP-ribose) polymerase [PARP] inhibitors have been used to treat breast and ovarian cancer susceptibility gene (BRCA) mutated tumors. Mechanisms of synthetic lethality targeting DNA repair defects.

DNA double strand breaks (DSBs) are one of the most serious types of DNA damage. DNA double strand break repair (DSBR) in cells can be roughly divided into three pathways, one is non-homologous end-joining (NHEJ), and the other is homologous recombination. , HR), and the last pathway is end-joining (alt-EJ) mediated by DNA polymerase θ (Polymerase θ, Polθ, POLQ) when NHEJ or HR is damaged, also known as microhomology-mediated end-joining Connect(MMEJ). Polθ plays a central role in microhomology-mediated end joining. Polθ has an N-terminal helicase domain and a C-terminal DNA polymerase domain. Studies have shown that the helicase domain of Polθ can promote the adhesion of microhomologies. After adhesion, all protruding bases are removed by nucleases and the gaps are filled by Polθ. Therefore, Polθ has received increasing attention as an important target of DNA repair defects. Polθ is barely expressed in normal tissues but is overrepresented in multiple tumor types (eg, breast, ovarian, HNSCC, and lung) and is associated with poor prognostic outcomes. Studies have shown that when homologous recombination-mediated repair is compromised (HRD), such as BRCA1 or BRCA2 mutations, Polθ is highly expressed and guides DSB repair toward microhomology-mediated end joining (MMEJ), initiating the DNA repair process of MMEJ. .

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

The purpose of the present invention is to provide a Polθ inhibitor, which has the structure of Formula I as shown, and can be used to inhibit the activity of Polθ polymerase and prevent or treat diseases or conditions mediated by Polθ.

The present invention provides a heterocyclic compound represented by formula I, its stereoisomer or its pharmaceutically acceptable salt: 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 ₂ are each independently halogen, hydroxyl, Amino group, cyano group, unsubstituted or C ₁ -C ₆ alkyl group substituted by one or more R _a , unsubstituted or C ₁ -C ₆ alkoxy group substituted by one or more R _a , unsubstituted or 3-6 membered cycloalkyl substituted by one or more R _a ; in R ₁ and R ₂ , each R _a is independently deuterium, halogen, hydroxyl, amine, cyano, C ₁ -C ₆ alkyl Or C ₁ -C ₆ haloalkyl; R ₃ is hydrogen, unsubstituted or C ₁ -C ₆ alkyl substituted by one or more R _a ; in R ₃ , each R _a is independently deuterium, halogen, hydroxyl _{or _ _ _ _ _ _ _ _ _ _} , R ₄ , R ₅ , R ₆ , R ₇ and R ₈ are each independently hydrogen or R _b , and at least one of R ₄ , R ₅ , R ₆ , R ₇ and R ₈ is a cyano group; or, R ₄ and R ₅ are each independently R _b , or R ₆ and R ₇ are each independently R _b , or R ₄ and R ₅ together with the C atom to which they are connected form ring A, R ₆ , R ₇ and R ₈ is each independently hydrogen or R _b ; or, R ₅ and R ₆ together with the C atom to which they are connected form ring A, and R ₄ , R ₇ and R ₈ are each independently hydrogen or R _b ; or, R ₆ and R ₇ together with the C atom to which they are connected form Ring A, R ₄ , R ₅ and R ₈ are each independently hydrogen or R _b ; or, R ₇ and R ₈ together with the C atom to which they are connected form Ring A , R ₄ , R ₅ and R ₆ are each independently hydrogen or R _b ; each R _b is independently halogen, hydroxyl, amino, cyano, unsubstituted or C ₁ -C ₆ alkyl substituted by R _a , unsubstituted or substituted by R _a C ₁ -C ₆ alkoxy group, unsubstituted or substituted by R _a 3-6 membered cycloalkyl group, unsubstituted or substituted by R _a 4-6 membered heterocycloalkyl group ; In each R _b , each R _a is independently deuterium, halogen, hydroxyl, amine, cyano, C ₁ -C ₆ alkyl or C ₁ -C ₆ haloalkyl; each ring A is not 3-8 membered cycloalkyl substituted or substituted by one or more R _a , 4-8 membered heterocycloalkyl unsubstituted or substituted by R _a ; In each ring A, the R _a is independently deuterium , halogen, hydroxyl, amino, cyano, C ₁ -C ₆ alkyl or C ₁ -C ₆ haloalkyl.

In a certain aspect, in the heterocyclic compound represented by formula I, its stereoisomer or a pharmaceutically acceptable salt thereof, certain groups in the heterocyclic compound represented by formula I are defined as follows: The definitions of the remaining groups are as described in any other scheme (hereinafter referred to as "in a certain scheme"): the heterocyclic compound shown in formula I, its stereoisomer or its pharmaceutically acceptable salt: 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 and R ₂ are each independently selected from: halogen, Hydroxy group, amino group, cyano group, unsubstituted or C ₁ -C ₆ alkyl group substituted by one or more R _a , unsubstituted or C ₁ -C ₆ alkoxy group substituted by one or more R _a , unsubstituted 3-6 membered cycloalkyl substituted or substituted by one or more R _a ; When m is 2, 3, 4, the R ₁ is the same or different; When n is 2, 3, 4, the R ₂ are the same or different; R ₃ is hydrogen, unsubstituted or C ₁ -C ₆ alkyl substituted by one or more Ra; R ₄ , R ₅ , R ₆ and R ₇ are each independently hydrogen or _{R b ,} R ₈ is R _b ; or R ₄ , R ₅ , R ₆ , R ₇ , and R ₈ are each independently hydrogen or R _b , and at least one of R ₄ , R ₅ , R ₆ , R ₇ , and R ₈ is cyanide. group; or R ₄ and R ₅ are the same or different R _b , or R ₄ and R ₅ together with the C atom to which they are connected form a ring A, and R ₆ , R ₇ , and R ₈ are each independently hydrogen or R _b ; or R ₅ and R ₆ together with the C atoms to which they are connected form ring A, and R ₄ , R ₇ , and R ₈ are each independently hydrogen or R _b ; or R ₆ and R ₇ are the same or different R _b , Or R ₆ and R ₇ together with the C atom to which they are connected form ring A, and R ₄ , R ₅ and R ₈ are each independently hydrogen or R _b ; or R ₇ and R ₈ together with the C atom to which they are connected form ring A. In Ring A, R ₄ , R ₅ and R ₆ are each independently hydrogen or R _b ; wherein any of the above R _b is each independently halogen, hydroxyl, amino, cyano, unsubstituted or substituted by R _a C ₁ -C ₆ alkyl, unsubstituted or R _a substituted C ₁ -C ₆ alkoxy, unsubstituted or R _a substituted 3-6 membered cycloalkyl, unsubstituted or R _a substituted 4-6 membered heterocycloalkyl; The ring A is a 3-8 membered cycloalkyl that is unsubstituted or substituted by one or more R _a , or a 4-8 membered heterocycloalkyl that is unsubstituted or substituted with R _a ; Any of the above R _a is independently deuterium, halogen, hydroxyl, amine group, cyano group, C ₁ -C ₆ alkyl group, C ₁ -C ₆ haloalkyl group; When R _a is multiple, the R _a is the same or different.

In a certain aspect, the heterocyclic compound represented by formula I, its stereoisomer or its pharmaceutically acceptable salt, 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 and R ₂ are each independently selected from: halogen, Hydroxy group, amino group, cyano group, unsubstituted or C ₁ -C ₆ alkyl group substituted by one or more R _a , unsubstituted or C ₁ -C ₆ alkoxy group substituted by one or more R _a , unsubstituted 3-6 membered cycloalkyl substituted or substituted by one or more R _a ; When m is 2, 3, 4, the R ₁ is the same or different; When n is 2, 3, 4, the R ₂ are the same or different; R ₃ is hydrogen, unsubstituted or C ₁ -C ₆ alkyl substituted by one or more Ra; R ₄ , R ₅ , R ₆ and R ₇ are each independently hydrogen or _{R b ,} R ₈ is R _b ; or R ₄ , R ₅ , R ₆ , R ₇ , and R ₈ are each independently hydrogen or R _b , and at least one of R ₄ , R ₅ , R ₆ , R ₇ , and R ₈ is cyanide. group; or R ₄ and R ₅ are the same or different R _b , or R ₄ and R ₅ together with the C atom to which they are connected form a ring A, and R ₆ , R ₇ , and R ₈ are each independently hydrogen or R _b ; or R ₅ and R ₆ together with the C atoms to which they are connected form ring A, and R ₄ , R ₇ , and R ₈ are each independently hydrogen or R _b ; or R ₆ and R ₇ are the same or different R _b , Or R ₆ and R ₇ together with the C atom to which they are connected form ring A, and R ₄ , R ₅ and R ₈ are each independently hydrogen or R _b ; or R ₇ and R ₈ together with the C atom to which they are connected form ring A. In Ring A, R ₄ , R ₅ and R ₆ are each independently hydrogen or R _b ; wherein any of the above R _b is each independently halogen, hydroxyl, amino, cyano, unsubstituted or substituted by R _a C ₁ -C ₆ alkyl, unsubstituted or R _a substituted C ₁ -C ₆ alkoxy, unsubstituted or R _a substituted 3-6 membered cycloalkyl, unsubstituted or R _a substituted 4-6 membered heterocycloalkyl; The ring A is a 3-8 membered cycloalkyl that is unsubstituted or substituted by one or more R _a , or a 4-8 membered heterocycloalkyl that is unsubstituted or substituted with R _a ; Any of the above R _a is each independently halogen, hydroxyl, amino group, cyano group, C ₁ -C ₆ alkyl group, C ₁ -C ₆ haloalkyl group; When R _a is multiple, the R _a Same or different.

In a certain solution, the ring K is a 5-6 membered N-containing heteroaromatic ring; preferably, the ring K is a pyridine ring, a pyrimidine ring, a pyrazine ring or a pyridazine ring.

In a certain scheme, the heterocyclic compound represented by formula I, its stereoisomer or its pharmaceutically acceptable salt, has structure II Wherein, m is 0, 1, 2, 3, 4; n is 0, 1, 2, 3, 4; R ₁ and R ₂ are each independently selected from: halogen, hydroxyl, amino group, cyano group, unsubstituted or C ₁ -C ₆ alkyl substituted by one or more R _a , C ₁ -C ₆ alkoxy unsubstituted or substituted by one or more R _a , unsubstituted or substituted by one or more R _a 3-6 membered cycloalkyl; When m is 2, 3, 4, the R ₁ is the same or different; When n is 2, 3, 4, the R ₂ is the same or different; R ₃ is hydrogen or un C ₁ -C ₆ alkyl substituted or substituted by one or more _Ra ; R ₄ , R ₅ , R ₆ , R ₇ are each independently hydrogen or R _b , R ₈ is R _b ; or R ₄ , R ₅ , R ₆ , R ₇ , and R ₈ are each independently hydrogen or R _b , and at least one of R ₄ , R ₅ , R ₆ , R ₇ , and R ₈ is a cyano group; or R ₄ and R ₅ are the same or Different R _b , or R ₄ and R ₅ together with the C atoms to which they are connected form ring A, R ₆ , R ₇ , R ₈ are each independently hydrogen or R _b ; or R ₅ and R ₆ are connected to them The C atoms together form ring A, and R ₄ , R ₇ , and R ₈ are each independently hydrogen or R _b ; or R ₆ and R ₇ are the same or different R _b , or R ₆ and R ₇ are connected to them. C atoms together form Ring A, R ₄ , R ₅ , R ₈ are each independently hydrogen or R _b ; or R ₇ and R ₈ together with the C atoms to which they are connected form Ring A, R ₄ , R ₅ , R ₆ Each is independently hydrogen or R _b ; wherein any of the above R _b is each independently halogen, hydroxyl, amino, cyano, unsubstituted or C ₁ -C ₆ alkyl substituted by R _a , unsubstituted Or a C ₁ -C ₆ alkoxy group substituted by R _a ; The ring A is a 3-8-membered cycloalkyl group that is unsubstituted or substituted by one or more R _a , or a 4-membered cycloalkyl group that is unsubstituted or substituted by R _a 8-membered heterocycloalkyl; any of the above R _a is independently halogen, hydroxyl, amino, cyano, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl; when R _a is multiple , the R _a is the same or different.

In a certain scheme, any of the above multiple R _a substitutions refers to 2, 3 or 4 R _a substitutions; when R _a is 2, 3 or 4, the R _a is the same or different.

In a certain aspect, the heterocyclic compound represented by formula II, its stereoisomer or its pharmaceutically acceptable salt: Wherein, m and n are respectively integers from 0 to 4; R ₁ and R ₂ are each independently selected from: halogen, hydroxyl, amino, cyano, C ₁ -C ₆ alkyl, 3-6 membered cycloalkyl; The R ₁ or R ₂ is optionally substituted with a substituent selected from the following: halogen, hydroxyl, amino group, C ₁ -C ₆ alkyl; when R ₁ is multiple, the R ₁ is the same or different; When R ₂ is multiple, the R ₂ is the same or different; R ₃ is hydrogen, C ₁ -C ₆ alkyl, C ₁ -C ₆ deuterated alkyl or C ₁ -C ₆ haloalkyl; R ₄ , R ₅ , R ₆ , and R ₇ are each independently hydrogen or C ₁ -C ₆ alkyl, and R ₈ is C ₁ -C ₆ alkyl; or R ₄ and R ₅ together with the C atom to which they are connected form Ring A. , R ₆ , R ₇ , and R ₈ are each independently hydrogen or C ₁ -C ₆ alkyl; or R ₅ and R ₆ together with the C atoms to which they are connected form ring A, and R ₄ , R ₇ , and R ₈ are each independently are independently hydrogen or C ₁ -C ₆ alkyl; or R ₆ and R ₇ together with the C atoms to which they are connected form ring A, and R ₄ , R ₅ and R ₈ are each independently hydrogen or C ₁ -C ₆ Alkyl; or R ₇ and R ₈ together with the C atoms to which they are connected form ring A, and R ₄ , R ₅ , and R ₆ are each independently hydrogen or C ₁ -C ₆ alkyl; 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 following: halogen, hydroxyl, amino, cyano, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl.

In a certain aspect, the heterocyclic compound represented by formula II, its stereoisomer or its pharmaceutically acceptable salt: m and n are respectively integers from 0 to 4; R ₁ and R ₂ are each independently selected From: halogen, hydroxyl, amino, cyano, C ₁ -C ₆ alkyl, 3-6 membered cycloalkyl; the R ₁ or R ₂ is optionally substituted with a substituent selected from the following: halogen, hydroxyl , amino group, C ₁ -C ₆ alkyl group; when R ₁ is multiple, the R ₁ is the same or different; when R ₂ is multiple, the R ₂ is the same or different; R ₃ is hydrogen, C ₁ -C ₆ alkyl or C ₁ -C ₆ haloalkyl; R ₄ , R ₅ , R ₆ and R ₇ are each independently hydrogen or C ₁ -C ₆ alkyl, R ₈ is C ₁ -C ₆ alkyl ; Or R ₄ and R ₅ together with the C atom to which they are connected form ring A, and R ₆ , R ₇ and R ₈ are each independently hydrogen or C ₁ -C ₆ alkyl; or R ₅ and R ₆ are together with the C atom to which they are connected. The connected C atoms together form Ring A, and R ₄ , R ₇ , and R ₈ are each independently hydrogen or C ₁ -C ₆ alkyl; or R ₆ and R ₇ together with the C atoms to which they are connected form Ring A, and R _4. R ₅ and R ₈ are each independently hydrogen or C ₁ -C ₆ alkyl; or R ₇ and R ₈ together with the C atoms to which they are connected form ring A, and R ₄ , R ₅ and R ₆ are each independently is hydrogen or C ₁ -C ₆ alkyl; the ring A is a 3-8-membered cycloalkyl or a 4-8-membered heterocycloalkyl; the ring A is optionally substituted with a substituent selected from the following: halogen , hydroxyl, amino, cyano, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl.

In a certain scheme, the heterocyclic compound represented by formula I, its stereoisomer or its pharmaceutically acceptable salt, has structure Ia or Ib Among them, m, n, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ and R ₈ are as defined in any one of the present invention.

In a certain plan, Have structure , R ₁₁ is C ₁ -C ₃ alkyl, R ₁₂ is C ₁ -C ₃ haloalkyl; preferably, Have structure .

In a certain plan, Have structure , R ₂₁ , R ₂₂ , and R ₂₃ are the same or different halogens; preferably, the R ₂₁ , R ₂₂ , and R ₂₃ are independently F or Cl; preferably, Have structure .

In a certain embodiment, R ₄ and R ₅ together with the C atom to which they are connected form Ring A, and R ₆ , R ₇ , and R ₈ are each independently hydrogen.

In a certain scheme, R ₄ and R ₅ together with the C atoms to which they are connected form ring A, and R ₆ , R ₇ , and R ₈ are each independently hydrogen, hydroxyl, C ₁ -C ₆ alkyl, C ₁ - C ₆ haloalkyl; or R ₅ and R ₆ together with the C atom to which they are connected form ring A, R ₄ , R ₇ , R ₈ are each independently hydrogen, hydroxyl, C ₁ -C ₆ alkyl, C ₁ - C ₆ haloalkyl; or R ₆ and R ₇ together with the C atom to which they are connected form ring A, R ₄ , R ₅ , R ₈ are each independently hydrogen, hydroxyl, C ₁ -C ₆ alkyl, C ₁ - C ₆ haloalkyl; or R ₇ and R ₈ together with the C atom to which they are connected form ring A, R ₄ , R ₅ , R ₆ are each independently hydrogen, hydroxyl, C ₁ -C ₆ alkyl, C ₁ - C ₆ haloalkyl.

In a certain scheme, R ₄ and R ₅ together with the C atom to which they are connected form ring A, and R ₆ , R ₇ and R ₈ are each independently hydrogen; or R ₅ and R ₆ and the C atom to which they are connected form ring A. Together they form Ring A, R ₄ , R ₇ , R ₈ are hydrogen; or R ₆ and R ₇ together with the C atom to which they are connected form Ring A, R ₄ , R ₅ , R ₈ are hydrogen; or R ₇ and R ₈ together with the C atoms to which they are connected form ring A, and R ₄ , R ₅ , and R ₆ are hydrogen; the ring A is a 3-6-membered cycloalkyl group or a 4-8-membered heterocycloalkyl group; Preferably, The ring A is selected from: cyclopropane, cyclobutane, cyclopentane, cyclohexane; Preferably, the 4-8 membered heterocycloalkyl group has 1, 2 or 3 selected from N, O or S of heteroatoms; when there are multiple heteroatoms, the heteroatoms are the same or different.

In a certain embodiment, R ₄ , R ₅ , R ₆ , and R ₇ are hydrogen, and R ₈ is C ₁ -C ₃ alkyl.

In a certain embodiment, the C ₁ -C ₆ haloalkyl group is a C ₁ -C ₆ fluoroalkyl group.

In a certain embodiment, the C ₁ -C ₃ haloalkyl group is C ₁ -C ₃ fluoroalkyl group.

In a certain scheme, R ₄ and R ₅ are the same or different R _b , R ₆ , R ₇ , and R ₈ are each independently hydrogen, hydroxyl, halogen, C ₁ -C ₆ alkyl, C ₁ -C ₆ Haloalkyl; or R ₆ and R ₇ are the same or different R _b , R ₄ , R ₅ and R ₈ are each independently hydrogen, hydroxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl; R _b is halogen, hydroxyl, cyano, C ₁ -C ₆ alkyl; preferably, R _b is fluorine, chlorine, hydroxyl, cyano, methyl, ethyl, propyl.

In a certain aspect, the heterocyclic compound represented by formula I, its stereoisomer or its pharmaceutically acceptable salt, wherein, has the structure Ic, Id or Ie , , ; Among them, ring K, m, n, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ and R ₈ are as defined in any one of the present invention.

In one embodiment, in Ic, _R5 and _R6 together with the C atom to which they are attached form Ring A.

In one embodiment, in Id, R ₅ and R ₆ together with the C atom to which they are attached form Ring A; or R ₆ and R ₇ together with the C atom to which they are attached form Ring A; or R ₇ and R ₈ together with the C atoms to which they are attached form Ring A.

In one embodiment, in Ie, R ₄ and R ₅ together with the C atom to which they are attached form Ring A; or R ₅ and R ₆ together with the C atom to which they are attached form Ring A.

In a certain solution, any of the above ring A is a 3-6-membered cycloalkyl or a 4-8-membered heterocycloalkyl; preferably, the ring A is selected from: cyclopropane, cyclobutane, cycloalkyl Pentane, cyclohexane.

In a certain aspect, the 4-8 membered heterocycloalkyl group has 1, 2 or 3 heteroatoms selected from N, O or S; when there are multiple heteroatoms, the heteroatoms are the same or different.

In one embodiment, _R3 is methyl.

In one embodiment, R ₃ is deuterated methyl (-CD ₃ ).

In a certain embodiment, in Ic, Id or Ie, R ₄ , R ₅ , R ₆ , R ₇ and R ₈ are each independently hydroxyl, methyl, fluoromethyl, cyano, cyclopropyl.

In a certain scheme, Ring K is a 5-membered or 6-membered N-containing heteroaromatic ring; preferably, Ring K is a pyridine ring, pyrimidine ring, pyrazine ring, or pyridazine ring.

In a certain plan, Have structure , , or .

In a certain aspect, m is 2 or 3, such as 2.

In a certain solution, n is 1 or 3, such as 3.

In a certain solution, the ring K is a 5- to 6-membered heteroaromatic ring. In the 5- to 6-membered heteroaromatic ring, the heteroatom is N and the number of heteroatoms is 1 or 2. For example, the ring K is pyridine.

In a certain scheme, each R ₁ is independently halogen, unsubstituted or C ₁ -C ₆ alkyl or 3-6 membered cycloalkyl substituted by one or more R _a . Preferably, each R ₁ is independently is independently halogen, unsubstituted or C ₁ -C ₆ alkyl substituted by one or more R _a , for example, each R ₁ is independently halogen, trifluoromethyl, methyl or trideuterated methyl, As another example, each R ₁ is independently a C ₁ -C ₆ alkyl group that is unsubstituted or substituted by one or more R _a .

In one embodiment, when m is 2, each _R1 is located in the ortho or para position to the heteroatom in ring K.

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

In a certain aspect, in each R ₁ , each R _a is independently a halogen or deuterium. For example, in each R ₁ , each R _a is independently a halogen.

In one embodiment, each _R2 is independently halogen.

In a certain embodiment, R ₃ is a C ₁ -C ₆ alkyl group that is unsubstituted or substituted by one or more R _a . For example, R ₃ is a C 1 -C 6 alkyl group, and for example, R ₃ is a C ₁ -C ₆ alkyl group substituted by one or more R a Multiple R _a substituted C ₁ -C ₆ alkyl groups.

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

In a certain scheme, each R _b is independently halogen, hydroxyl, cyano, 3-6 membered cycloalkyl or C ₁ -C ₆ alkyl that is unsubstituted or substituted by one or more R _a , preferably , each R _b is independently halogen, hydroxyl or C ₁ -C ₆ alkyl. For example, each R _b is independently hydroxyl or C ₁ -C ₆ alkyl.

In one aspect, each R _a in each R _b is independently a halogen.

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

In one embodiment, R ₄ , R ₅ , R ₆ and R ₇ are each independently hydrogen or R _b and R ₈ is C ₁ -C ₆ alkyl.

In one embodiment, R ₅ , R ₆ , R ₇ and R ₈ are each independently hydrogen or R _b and R ₄ is cyano.

In one embodiment, R ₄ , R ₅ , R ₇ and R ₈ are each independently hydrogen or R _b and R ₆ is cyano.

In one embodiment, R ₄ and R ₅ are each independently R _b and R ₆ , R ₇ and R ₈ are each independently R _b or hydrogen, for example, R ₄ and R ₅ are each independently R _b , R ₆ and R ₇ are each independently hydrogen, C ₁ -C ₆ alkyl or hydroxyl, R ₈ is hydrogen, preferably, R ₄ and R ₅ are each independently R _b , R ₆ and R ₇ are each independently hydrogen Or hydroxyl, R ₈ is hydrogen.

In one embodiment, R ₆ and R ₇ are each independently R _b and R ₄ , R ₅ and R ₈ are each independently R _b or hydrogen, for example, R ₆ and R ₇ are each independently R _b , R ₄ and R ₅ are each independently hydrogen, C ₁ -C ₆ alkyl or hydroxyl, and R ₈ is hydrogen.

In a certain embodiment, R ₄ and R ₅ together with the C atom to which they are attached form Ring A, R ₆ and R ₇ are each independently hydrogen or hydroxyl, and R ₈ is hydrogen, for example, R ₄ and R ₅ are The attached C atoms together form Ring A, R ₆ and R ₇ are each independently hydrogen, and R ₈ is hydrogen.

In one embodiment, R ₅ and R ₆ together with the C atom to which they are attached form Ring A, R ₄ and R ₇ are each independently hydrogen or hydroxyl, and R ₈ is hydrogen.

In one embodiment, R ₆ and R ₇ together with the C atom to which they are attached form Ring A, R ₄ and R ₅ are each independently hydrogen or hydroxyl, and R ₈ is hydrogen.

In a certain scheme, in ring K, in the 5-6 membered heteroaromatic ring, the heteroatoms are selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3, for example, ring K , the 5-6 membered heteroaromatic ring is a 6-membered heteroaromatic ring. Preferably, in the 5-6 membered heteroaromatic ring, the heteroatom is selected from N. More preferably, the 5-6 membered heteroaromatic ring is The number of heteroatoms in the aromatic ring is 1.

In a certain embodiment, in R ₁ and R ₂ , the halogen is each independently fluorine, chlorine, bromine or iodine, such as chlorine or fluorine.

In a certain scheme, among R ₁ and R ₂ , the C 1 -C ₆ alkyl groups in the unsubstituted or substituted C _{1 -C 6} alkyl groups by one or more R _a are each independently methyl, ethyl, etc. base, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, such as methyl.

In a certain scheme, among R ₁ and R ₂ , the C 1 -C ₆ alkoxy groups in the unsubstituted or substituted C _{1 -C 6} alkoxy groups by one or more R _a are each independently methoxy. base, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy or tert-butoxy.

In a certain embodiment, in R ₁ and R ₂ , each of the 3-6 membered cycloalkyl groups is independently a cyclopropyl group, a cyclobutanyl group, a cyclopentyl group or a cyclohexyl group, such as a cyclopropyl group.

In a certain embodiment, in each of R ₁ and _{R 2} , the halogen is independently fluorine, chlorine, bromine or iodine, such as fluorine.

In a certain scheme, among R ₁ and R ₂ , in each R _a , the C ₁ -C ₆ alkyl groups are each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, iso- Butyl or tertiary butyl.

In a certain embodiment, in R ₁ and R ₂ , in each R _a , the C ₁ -C ₆ haloalkyl group is each independently 1, 2 or 3 halogens (such as fluorine, chlorine, bromine or iodine) Substituted C ₁ -C ₆ alkyl (eg methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl).

In a certain scheme, in R ₃ , the C ₁ -C ₆ alkyl groups in the unsubstituted or substituted C ₁ -C ₆ alkyl groups by one or more R _a are each independently methyl, ethyl, n- Propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl.

In a certain embodiment, in _R3 , in each _Ra , the halogen is independently fluorine, chlorine, bromine or iodine, such as fluorine.

In a certain scheme, in R ₃ , in each R _a , the C ₁ -C ₆ alkyl groups are each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or 3rd butyl.

In a certain scheme, in R ₃ , in each R _a , the C ₁ -C ₆ haloalkyl groups are each independently substituted with 1, 2 or 3 halogens (such as fluorine, chlorine, bromine or iodine) C ₁ -C ₆ alkyl (eg methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl).

In a certain scheme, among R ₄ , R ₅ , R ₆ , R ₇ and R ₈ , the C ₁ -C ₆ alkyl groups are each independently methyl, ethyl, n-propyl, isopropyl, n-propyl, etc. Butyl, isobutyl or tert-butyl, for example methyl.

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

In a certain scheme, in each R _b , the C ₁ -C ₆ alkyl groups in the unsubstituted or substituted C ₁ -C ₆ alkyl groups are each independently methyl _, ethyl, n-propyl, Isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl.

In a certain scheme, in each R _b , the C ₁ -C ₆ alkoxy group in the unsubstituted or substituted C ₁ -C ₆ alkoxy group by R _a is each independently a methoxy group, an ethoxy group, n-propoxy, isopropoxy, n-butoxy, isobutoxy or tert-butoxy.

In a certain scheme, in each R _b , the 3-6-membered cycloalkyl groups in the unsubstituted or substituted 3-6-membered cycloalkyl groups by R _a are each independently a cyclopropyl group, a cyclobutyl group, a cycloalkyl group, Pentyl or cyclohexyl, such as cyclopropyl.

In a certain scheme, in each R _b , in the 4-6 membered heterocycloalkyl group that is unsubstituted or substituted by R _a , the heteroatom in the 4-6 membered heterocycloalkyl group is selected from N, O and S. One or more types, the number of heteroatoms is 1, 2 or 3.

In a certain embodiment, in each R _b and each R _a , the halogen is each independently fluorine, chlorine, bromine or iodine, such as fluorine.

In a certain scheme, in each R _b , in each R _a , the C ₁ -C ₆ alkyl groups are each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl Or tertiary butyl.

In a certain scheme, in each R _b and each R _a , the C ₁ -C ₆ haloalkyl groups are each independently substituted by 1, 2 or 3 halogens (such as fluorine, chlorine, bromine or iodine). C ₁ -C ₆ alkyl (eg methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl).

In a certain scheme, in each ring A, the 3-8-membered cycloalkyl group in the 3-8-membered cycloalkyl group that is unsubstituted or substituted by one or more R _a is a 3-6-membered cycloalkyl group, for example Each of the 3-6 membered cycloalkyl groups is independently a cyclopropyl group, a cyclobutyl group, a cyclopentyl group or a cyclohexyl group, preferably a cyclopropyl group.

In a certain scheme, in each ring A, the 4-8-membered heterocycloalkyl group in the unsubstituted or R _a- substituted 4-8-membered heterocycloalkyl group is a 4-6-membered heterocycloalkyl group, preferably Preferably, in the 4-6 membered heterocycloalkyl group, the heteroatoms are selected from one or more types of N, O and S, and the number of heteroatoms is 1, 2 or 3.

In a certain embodiment, in each ring A and each R _a , the halogen is independently fluorine, chlorine, bromine or iodine.

In a certain scheme, in each ring A, in each R _a , the C ₁ -C ₆ alkyl groups are each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl Or tertiary butyl.

In a certain scheme, in each ring A and each R _a , the C ₁ -C ₆ haloalkyl groups are each independently substituted by 1, 2 or 3 halogens (such as fluorine, chlorine, bromine or iodine). C ₁ -C ₆ alkyl (eg methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl).

In a certain solution, when m is 2, 3, or 4, the R ₁ 's are the same or different.

In a certain solution, when n is 2, 3, or 4, the R ₂ is the same or different.

In a certain solution, when there are multiple R _a's , the R _a's may be the same or different.

In a certain solution, the "or" is only "or", expressing an alternative relationship between elements.

In a certain embodiment, when the alkyl group does not indicate substitution, the alkyl group is an unsubstituted alkyl group.

In a certain aspect, when the alkyl group is not stated to be isotopically substituted, the alkyl group is an alkyl group that does not contain carbon and/or hydrogen isotopes.

In a certain aspect, when the cycloalkyl group or carbocyclyl group is not specified as an unsaturated or partially saturated carbocyclic ring, the cycloalkyl group or carbocyclyl group is a saturated cycloalkyl group or carbocyclyl group.

In a certain aspect, when the heterocycloalkyl group is not specified as an unsaturated or partially saturated heterocycloalkyl group, the heterocycloalkyl group is a saturated heterocycloalkyl group.

In a certain aspect, when the heteroaromatic ring group is not specified as a heteroaromatic ring group condensed with a benzene ring, the heteroaromatic ring group is an unfused heteroaromatic ring group.

In a certain aspect, the stereoisomer is a paraporal isomer or a non-paraporal isomer.

In a certain aspect, in the heterocyclic compound represented by Formula I, its stereoisomer or its pharmaceutically acceptable salt, m is 0, 1, 2, 3 or 4; n is 0, 1, 2 , 3 or 4; Ring K is a 5- to 6-membered heteroaromatic ring. In the 5- to 6-membered heteroaromatic ring, the heteroatom is N, and the number of heteroatoms is 1 or 2; each R ₁ is independently halogen, C ₁ -C ₆ alkyl or 3-6 membered cycloalkyl that is unsubstituted or substituted by one or more R _a ; In each R ₁ , each R _a is independently halogen or deuterium; R ₂ is each independently Halogen; R ₃ is a C ₁ -C ₆ alkyl group that is unsubstituted or substituted by one or more R _a ; in R ₃ , each R _a is independently deuterium; R ₄ , R ₅ , R ₆ and R ₇ are each are independently hydrogen or R _b , R ₈ is C ₁ -C ₆ alkyl; or, R ₅ , R ₆ , R ₇ and R ₈ are each independently hydrogen or R _b , and R ₄ is cyano; or, R ₄ , R ₅ , R ₇ and R ₈ are each independently hydrogen or R _b , and R ₆ is cyano; or, R ₄ and R ₅ are each independently R _b , R ₆ and R ₇ are each independently R Hydrogen, C ₁ -C ₆ alkyl or hydroxyl, R ₈ is hydrogen; or, R ₆ and R ₇ are each independently R _b , R ₄ and R ₅ are each independently hydrogen, C ₁ -C ₆ alkyl or Hydroxy, R ₈ is hydrogen; or, R ₄ and R ₅ together with the C atom to which they are connected form ring A, R ₆ and R ₇ are each independently hydrogen or hydroxyl, R ₈ is hydrogen; or, R ₅ and R ₆ together with the C atom to which they are connected form Ring A, R ₄ and R ₇ are each independently hydrogen or hydroxyl, and R ₈ is hydrogen; or, R ₆ and R ₇ together with the C atom to which they are connected form Ring A, R ₄ and R ₅ are each independently hydrogen or hydroxyl, R ₈ is hydrogen; each R _b is independently halogen, hydroxyl, cyano, 3-6 membered cycloalkyl or unsubstituted or by one or more R _a Substituted C ₁ -C ₆ alkyl; in each R _b , each R _a is independently a halogen; each ring A is independently a 3-8 membered cycloalkyl group.

In a certain aspect, in the heterocyclic compound represented by formula I, its stereoisomer or its pharmaceutically acceptable salt, m is 2 or 3; n is 3; ring K is pyridine; each R ₁ is respectively Independently halogen, unsubstituted or C ₁ -C ₆ alkyl substituted by one or more R _a ; When m is 2, each R ₁ is located in the ortho or para position of the heteroatom in ring K, when m is 3, each R ₁ is adjacent and located in the ortho, meta or para position of the heteroatom in ring K; In each R ₁ , each R _a is independently halogen or deuterium; R ₂ is each independently halogen; R ₃ is a C ₁ -C ₆ alkyl group that is unsubstituted or substituted by one or more R _a ; in R ₃ , each R _a is independently deuterium; R ₄ and R ₅ are each independently R _b , R ₆ and R ₇ is each independently hydrogen or hydroxyl, and R ₈ is hydrogen; or, R ₄ and R ₅ together with the C atom to which they are connected form ring A, R ₆ and R ₇ are each independently hydrogen, and R ₈ is hydrogen; Each R _b is independently halogen, hydroxyl or C ₁ -C ₆ alkyl; each ring A is independently a 3-8 membered cycloalkyl.

In a certain aspect, in the heterocyclic compound represented by formula I, its stereoisomer or its pharmaceutically acceptable salt, m is 2 or 3; n is 3; ring K is pyridine; each R ₁ is respectively Independently halogen, trifluoromethyl, methyl or trideuterated methyl; when m is 2, each R ₁ is located at the ortho or para position of the heteroatom in ring K, when m is 3, each R ₁ Adjacent and located in the ortho, meta or para position of the heteroatom in ring K; R ₂ is each independently halogen; R ₃ is a C ₁ -C ₆ alkyl group that is unsubstituted or substituted by one or more R _a ; In R ₃ , each _Ra is independently deuterium; R ₄ and R ₅ are each independently R _b , R ₆ and R ₇ are each independently hydrogen or hydroxyl, and R ₈ is hydrogen; or, R ₄ and R ₅ Together with the C atoms to which they are connected, they form ring A, R ₆ and R ₇ are each independently hydrogen, and R ₈ is hydrogen; each R _b is independently hydroxyl or C ₁ -C ₆ alkyl; each ring A is independently Ground is a 3-8 membered cycloalkyl group.

In a certain aspect, in the heterocyclic compound represented by formula I, its stereoisomer or its pharmaceutically acceptable salt, m is 2; n is 3; ring K is pyridine; each R ₁ is independently is a C ₁ -C ₆ alkyl group that is unsubstituted or substituted by one or more R _a ; when m is 2, each R ₁ is located in the ortho or para position of the heteroatom in ring K; in each R ₁ , each R _a is each independently halogen; R ₂ is each independently halogen; R ₃ is C ₁ -C ₆ alkyl; R ₄ and R ₅ are each independently R _b , and R ₆ and R ₇ are each independently hydrogen or hydroxyl , R ₈ is hydrogen; or, R ₄ and R ₅ form ring A together with the C atom to which they are connected, R ₆ and R ₇ are each independently hydrogen, and R ₈ is hydrogen; each R _b is independently hydroxyl or C ₁ -C ₆ alkyl; each ring A is independently a 3-8 membered cycloalkyl group.

In a certain aspect, in the heterocyclic compound represented by formula I, its stereoisomer or its pharmaceutically acceptable salt, m is 2 or 3; n is 3; ring K is pyridine; each R ₁ is respectively Independently halogen, trifluoromethyl, methyl or trideuterated methyl; when m is 2, each R ₁ is located at the ortho or para position of the heteroatom in ring K, when m is 3, each R ₁ Adjacent and located in the ortho, meta or para position of the heteroatom in ring K; In each R ₁ , each R _a is independently halogen or deuterium; R ₂ is each independently halogen; R ₃ is substituted by one or more R _a substituted C ₁ -C ₆ alkyl; in R ₃ , each R _a is independently deuterium; R ₄ and R ₅ are each independently R _b , and R ₆ and R ₇ are each independently hydrogen or hydroxyl , R ₈ is hydrogen; each R _b is independently a hydroxyl group or a C ₁ -C ₆ alkyl group; each ring A is independently a 3-8 membered cycloalkyl group.

In a certain plan, for , , , , , , or .

In a certain plan, for , , , , , , , , , , , , , , or .

In a certain plan, for or .

In a certain plan, for , , or .

In one embodiment, Ring K is pyridine or pyrimidine.

In one embodiment, _R1 is methyl, trifluoromethyl ( _-CF3 ), difluoromethyl ( _-CHF2 ), cyclopropyl, fluoro, or trideuteratedmethyl ( _-CD3 ).

In one embodiment, _R2 is fluorine or chlorine.

In one embodiment, R ₃ is methyl or trideuterated methyl (-CD ₃ ).

In one embodiment, R ₄ and R ₅ are each independently hydrogen, fluorine, hydroxyl, methyl, cyclopropyl, trifluoromethyl, cyano, or difluoromethyl.

In one embodiment, R ₄ and R ₅ together with the C atom to which they are attached form Ring A, which is cyclopropanyl.

In one embodiment, R ₅ and R ₆ together with the C atom to which they are attached form Ring A, which is cyclopropanyl.

In one embodiment, R ₆ and R ₇ are each independently hydrogen, hydroxyl, methyl or cyano.

In one embodiment, R ₈ is hydrogen or methyl.

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

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

The present invention provides a pharmaceutical composition, including the heterocyclic compound represented by the above formula I, its stereoisomer or its pharmaceutically acceptable salt and optionally i) one or more active drugs; and/or ii) Pharmaceutically acceptable carrier.

The present invention provides a pharmaceutical composition, including a heterocyclic compound represented by the above formula I, its stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and/or diluent.

The present invention provides a heterocyclic compound represented by the above formula I, its stereoisomer or a pharmaceutically acceptable salt thereof, or the use of the above pharmaceutical composition, including: 1) Inhibit Polθ activity; 2) Prevent and treat Polθ-mediated diseases; 3) Preparation of drugs, pharmaceutical compositions or preparations for inhibiting Polθ activity; 4) Preparation of drugs, pharmaceutical compositions or preparations for preventing and treating Polθ-mediated diseases; Preferably, the Polθ-mediated disease is cancer.

The present invention provides a heterocyclic compound represented by the above formula I, its stereoisomer or a pharmaceutically acceptable salt thereof, or the above pharmaceutical composition for preventing or treating cancer.

The present invention also provides a method for treating diseases, which includes administering to a patient a therapeutically effective amount of at least one of the heterocyclic compound represented by formula I, its stereoisomer or its pharmaceutically acceptable salt, or the above pharmaceutical composition.

In one embodiment, the disease is a Polθ-mediated disease, such as a disease characterized by overexpression of Rolq.

In a certain aspect, the disease is a cell proliferative disorder; preferably, the cell proliferative disorder is cancer.

In a certain aspect, the patient is a mammal, preferably a human.

In a certain scenario, examples of cancers (and their benign counterparts) that can be treated (or inhibited) by the heterocyclic compound represented by Formula I include, but are not limited to: tumors of epithelial origin (adenomas and various types of cancers, including adenocarcinoma, squamous carcinoma, transitional cell carcinoma and other cancers) such as bladder and urinary tract cancers, breast cancer, gastrointestinal cancers (including esophageal cancer, gastric (stomach) cancer, small bowel cancer, colon cancer, rectal cancer, and anal cancer) , liver cancer (hepatocellular carcinoma), gallbladder and biliary system cancer, exocrine pancreatic cancer, kidney, lung (such as adenocarcinoma, small cell lung cancer, non-small cell lung cancer, bronchioloalveolar carcinoma, and mesothelioma), head and neck cancer (such as Tongue cancer, oral cavity cancer, laryngeal cancer, pharyngeal cancer, nasopharyngeal cancer, tonsil cancer, salivary gland cancer, nasal cavity and sinus), reproductive system (such as ovaries, fallopian tubes, peritoneum, vagina, vulva, penis, cervix, myometrium, Endometrial) cancer, thyroid cancer (e.g., follicular thyroid carcinoma), adrenal cancer, prostate cancer, skin cancer (e.g., melanoma, basal cell carcinoma, squamous cell carcinoma, keratoacanthoma, dysplastic nevus); blood system Malignant neoplasms (i.e., leukemias, lymphomas) and premalignant hematologic and borderline malignant neoplastic diseases, including hematologic malignancies and related lymphoid disorders (e.g., acute lymphoblastic leukemia, chronic lymphocytic leukemia, B-cell lymphoma, e.g. Diffuse large B-cell lymphoma, follicular lymphoma, Burkitt lymphoma, mantle cell lymphoma, MALT lymphoma, T-cell lymphoma and leukemia, natural killer cell lymphoma, Hodgkin lymphoma, hairy cell leukemia, monoclonal gammopathy of undetermined significance, plasmacytoma, multiple myeloma, and post-transplant lymphoproliferative disorders), as well as hematological malignancies and myeloid-related diseases (e.g., acute myeloid leukemia, chronic myeloid leukemia, chronic myelomonocytic leukemia, eosinophilia, myeloproliferative disorders such as polycythemia vera), essential thrombocythemia and primary myelofibrosis, myeloproliferative syndromes, myelodysplasia syndrome and promyelocytic leukemia); tumors of mesenchymal origin, such as sarcomas of soft tissue, bone, or cartilage, such as osteosarcoma, fibrosarcoma, chondrosarcoma, rhabdomyosarcoma, leiomyosarcoma, liposarcoma, angiosarcoma, kaposi Sarcoma, Ewing's sarcoma, gastrointestinal epithelial sarcoma, epithelial sarcoma, malignant epithelial sarcoma histiocytoma, and dermatofibrosarcoma protuberans; central or peripheral nervous system tumors (such as astrocytomas, gliomas and glioblastomas, meningiomas , ependymomas, pineal tumors, and schwannomas); endocrine tumors (such as pituitary tumors, adrenal tumors, islet cell tumors, parathyroid tumors, carcinoid tumors, and medullary thyroid carcinoma); eye and adnexal tumors Neoplasms (such as retinoblastoma); germ cell and trophoblastic tumors (such as teratomas, seminoma, dysgerminoma, mole, and choriocarcinoma); and pediatric and embryonal tumors (such as medulloblastoma , neuroblastoma, Wilms tumor, and primitive neuroectodermal tumors); or congenital or otherwise predisposing the patient to malignancy (e.g., xeroderma pigmentosum).

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 invention 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 invention.

Definition of terms:

Unless otherwise stated, the definitions of groups and terms recorded in the specification and patent scope of this application include their definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, and definitions of specific compounds in the examples. etc., can be arbitrarily combined and combined with each other. Such combinations and combined group definitions and compound structures should fall within the scope described in the specification of this application.

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

It should be understood that the foregoing brief description and the following detailed description are exemplary and are for explanation only, and do not limit the subject matter of the present invention in any way. In this application, the use of the singular includes the plural unless specifically stated otherwise. It must be noted that, unless the context clearly dictates otherwise, the singular form used in this specification and claims includes the plural form of the referent. It should also be noted that the use of "or", "or" means "and/or" unless stated otherwise. Furthermore, the use of the term "includes" and other forms such as "includes," "contains," and "contains" is not limiting.

Unless otherwise stated, conventional methods within the skill in the art, such as mass spectrometry, NMR, IR and UV/VIS spectroscopy and pharmacological methods, were used. Unless specific definitions are given, the terminology employed herein in connection with the description of analytical chemistry, synthetic organic chemistry, and pharmaceutical and medicinal chemistry is known in the art. Standard techniques may be used in chemical synthesis, chemical analysis, drug preparation, formulation and delivery, and treatment of patients. For example, the manufacturer's instructions for using the kit can be used, or the reaction and purification can be carried out in accordance with methods known in the art or the instructions of the present invention. The above techniques and methods can generally be implemented in accordance with conventional methods well known in the art, as described in the various general and more specific documents cited and discussed in this specification. In this specification, groups and their substituents may be selected by those skilled in the art to provide stable moieties and compounds.

When a substituent is described by a conventional chemical formula written from left to right, the substituent also includes substituents that are chemically equivalent when the structural formula is written from right to left. For example, CH ₂ O is equivalent to OCH ₂ . As used in this article, or Indicates the attachment site of the group.

The section headings used in this article are for the purpose of organizing the article only and should not be construed as limitations on the subject matter described. All documents or portions of documents cited in this application, including but not limited to patents, patent applications, articles, books, operating manuals, and theses, are hereby incorporated by reference in their entirety.

In addition to the foregoing, when used in the description and patent scope of this application, the following terms have the meanings shown below unless otherwise specified.

When the numerical range described in the specification and patent scope of this application is understood as an "integer", it should be understood that the two endpoints of the range and each integer within the range are recorded. For example, "integers from 1 to 6" should be understood as describing each integer of 1, 2, 3, 4, 5, and 6.

In this application, "Polθ", "POLQ", "Polθ polymerase" and "Polθ protein polymerase" have the same definitions.

In this application, the term "halogen" means fluorine, chlorine, bromine, iodine, alone or as part of another substituent.

As used herein, the term "alkyl" when used alone or as part of other substituents means consisting only of carbon atoms and hydrogen atoms, containing no unsaturated bonds, having, for example, 1 to 6 carbon atoms and being connected to the molecule by a single bond. The remainder is connected to a linear or branched hydrocarbon chain group. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl, isopentyl, neopentyl and hexyl. Alkyl groups may be unsubstituted or substituted with one or more suitable substituents. Alkyl groups may also be isotopomers of naturally abundant alkyl groups that are rich in isotopes of carbon and/or hydrogen (i.e., deuterium or tritium).

The term "C ₁ -C ₆ alkyl" when used alone or as part of other substituents 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 group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, second butyl, third butyl, isopentyl, 2-methylbutyl base, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methyl Pentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl base, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, etc. or their isomers. In particular, said groups have 1, 2 or 3 carbon atoms ("C ₁ -C ₃ alkyl"), for example methyl, ethyl, n-propyl or isopropyl.

"Haloalkyl" when used alone or as part of another substituent is intended to include 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, in which one or more hydrogen atoms are substituted by halogen. Examples of haloalkyl groups include, but are not limited to, trifluoromethyl, trichloromethyl, pentafluoroethyl, pentachloroethyl, 2,2,2-trifluoroethyl, heptafluoropropyl, and heptachloropropyl.

The term "C ₁ -C ₆ alkoxy" when used alone or as part of other substituents is understood to mean a straight or branched saturated monovalent hydrocarbon radical having 1, 2, 3, 4, 5 or 6 carbon atoms. and oxygen atoms, or expressed as C ₁ -C ₆ alkyl -OC ₁ -C ₆ alkyl is defined as described in this specification, the oxygen atom can be attached to the straight or straight chain of the C ₁ -C ₆ alkyl group on any 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 "mn-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 single ring, two rings, three rings, spiro rings or bridged rings. Examples of unsubstituted cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Cycloalkyl groups may be substituted by one or more substituents.

The term "heterocycle" or "heterocycloalkyl" or "heterocyclyl" when used alone or as part of other substituents means one or more (in some embodiments 1 to 3) carbon atoms are replaced by Cycloalkyl substituted 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, a heterocycloalkyl group can be a heterocycloalkyl group fused to an aryl or heteroaryl group. When an initial number such as a 3-8 member is used to represent a heterocycloalkyl group, the number of carbons is also meant to include heteroatoms. Including single ring, two rings, three rings, spiro rings or bridged rings. Non-limiting examples of heterocycles include, but are not limited to, dihydropyridazine, dihydropyrazine, including substituted forms thereof, such as heterocyclyl including, but not limited to, 6-oxo-1,6-dihydropyridazine- 3-yl, 4-methyl-5-oxo-4,5-dihydropyrazin-2-yl, etc.

The term "heteroaryl" or "heteroaryl" when used alone or as part of other substituents refers to a monocyclic or polycyclic aromatic ring system, and in certain embodiments, 1 to 3 atoms in the ring system are Heteroatoms are elements other than carbon, including but not limited to N, O, S or P. Examples include furyl, imidazolyl, indolyl, pyrrolidinyl, pyrimidinyl, tetrazolyl, thienyl, pyridyl, pyrrolyl, N-methylpyrrolyl, quinolyl and isoquinolyl. The heteroaromatic ring group may be optionally condensed with a benzene ring, and may also include a monocyclic ring, a bicyclic ring, a tricyclic ring, a spiro ring or a bridged ring.

When used alone or as part of other substituents, "5-6 membered heteroaryl" is understood to have 5-6 ring atoms - and contain 1 or more heteroatoms independently selected from N, O and S of aromatic ring groups. Preferably 1 to 3 - aromatic ring groups of heteroatoms independently selected from N, O and S. In particular, the heteroaromatic ring group is selected from the group consisting of thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, Thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, etc.

In this application, the term "salt" or "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts. The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and/or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with human and animal tissue without undue toxicity, irritation, allergic reactions, or other problems or complications, commensurate with a reasonable benefit/risk ratio.

"Pharmaceutically acceptable acid addition salts" refer to salts formed with inorganic or organic acids that retain the biological effectiveness of the free base without other side effects. "Pharmaceutically acceptable base addition salts" refer to salts formed with inorganic or organic bases that can maintain the biological effectiveness of the free acid without other side effects. In addition to pharmaceutically acceptable salts, other salts are contemplated by the present invention. 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 invention.

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

The term "stereoisomers" refers to isomers resulting from different spatial arrangements of atoms in the molecule, including cis-trans isomers, parapolar isomers, non-parapolar isomers and conformational isomers. .

Depending on the choice of starting materials and methods, the compounds of the invention may exist in the form of one of the possible isomers or a mixture thereof, for example as pure optical isomers, or as a mixture of isomers, for example as racemic and non-racemic isomers. For mixtures of isomers, this depends on the number of asymmetric carbon atoms. When describing optically active compounds, the initial codes D and L or R and S are used to indicate the absolute configuration of the molecule with respect to the chiral center (or centers) in the molecule. The first codes D and L or (+) and (–) are symbols used to specify the rotation of plane polarized light caused by the compound, where (–) or L indicates that the compound is levorotatory. Compounds starting with (+) or D are dextrorotatory.

When the bond with the chiral carbon in the formula of the present invention is described as a straight line, it should be understood that the two configurations of the chiral carbon (R) and (S) and the resulting chiral pure compound and Both mixtures are included within the scope of this general formula. Use wedge-shaped bonds and dashed-line bonds to represent the absolute configuration of a stereocenter.

As used herein, "pharmaceutical composition" refers to a formulation of a compound of the present invention with a vehicle generally accepted in the art for delivering a biologically active compound to a mammal, such as a human. The medium includes a pharmaceutically acceptable carrier. The purpose of pharmaceutical compositions is to promote the administration of drugs to organisms and facilitate the absorption of active ingredients to exert biological activity.

In this application, "pharmaceutically acceptable carrier" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener approved by the relevant government regulatory authorities as acceptable for human or livestock use , diluents, preservatives, dyes/colorants, flavoring agents, surfactants, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, solvents or emulsifiers.

The compounds of the present invention may contain unnatural proportions of atomic isotopes on one or more of the atoms that make up the compound. For example, compounds can be labeled with radioactive isotopes such as deuterium ( ² H), tritium ( ³ H), iodine-125 ( ¹²⁵ I) or C-14 ( ¹⁴ C). All variations in the isotopic composition of the compounds of the invention, whether radioactive or not, are included within the scope of the invention.

The term "excipient" means a pharmaceutically acceptable inert ingredient. Examples of types of the term "excipient" include, but are not limited to, binders, disintegrants, lubricants, glidants, stabilizers, fillers, diluents, and the like. Excipients can enhance the handling properties of pharmaceutical formulations, i.e. make the formulation more suitable for direct compression by increasing flowability and/or viscosity.

As used herein, the term "treatment" and other similar synonyms include the following meanings: (i) prevent the occurrence of a disease or condition in mammals, particularly where such mammals are susceptible to the disease or condition but have not yet been diagnosed as having the disease or condition; (ii) inhibit a disease or condition, that is, arrest its progression; or (iii) alleviate a disease or condition, that is, cause the condition of the disease or condition to subside; or (iv) Reduce the symptoms of the disease or condition.

In the reaction of each step, the reaction temperature can be appropriately selected based on the solvent, starting materials, reagents, etc., and the reaction time can also be appropriately selected based on the reaction temperature, solvent, starting materials, reagents, etc. After the reaction of each step is completed, the target compound can be separated and purified from the reaction system according to common methods, such as filtration, extraction, recrystallization, washing, silica gel column chromatography and other methods. Without affecting the next step of the reaction, the target compound can also directly enter the next step of the reaction without separation and purification.

On the basis of not violating common sense in the field, the above preferred conditions can be combined arbitrarily to obtain preferred examples of the present invention.

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

The positive progressive effect of the present invention is that the inventor unexpectedly developed a Polθ inhibitor after extensive and in-depth research. The Polθ inhibitor has the structure shown in the Chinese formula I of the present invention and has a good inhibitory effect on Polθ polymerase. It can prevent or treat diseases or conditions mediated by Polθ, exhibits excellent pharmacokinetic properties, and has high safety and pharmaceutical properties.

The present invention is further described below by means of examples, but the present invention is not limited to the scope of the described examples. Experimental methods that do not indicate specific conditions in the following examples should be selected according to conventional methods and conditions, or according to product specifications.

Example 1 Preparation of Compound I-1A

The synthesis route is as follows:

Step 1: Preparation of 1-(tert-butyl)2-ethyl( R )-5-oxopyrrolidine-1,2-dicarboxylate (B1-2)

Dissolve ( R )-5-oxopyrrolidine-2-carboxylic acid ethyl ester (10.0 g, 63.6 mmol) and di-tert-butyl dicarbonate (16.66 g, 76 mmol) in tetrahydrofuran (100 mL), 0°C Then add N,N -dimethylpyridin-4-amine (0.777 g, 6.36 mmol), and the reaction solution is stirred at 25°C for 10 hours. After the reaction is completed, concentrate the reaction solution, add water (20 mL) to dissolve, and extract with ethyl acetate (20 mL*3). Combine the organic layers, wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, and concentrate to obtain the crude product. 1-(tert-butyl)2-ethyl( R )-5-oxopyrrolidine-1,2-dicarboxylate (13.9 g, yield 85%).

LC-MS, M/Z (ESI): 258.2 [M+H] ⁺ .

Step 2: 1-(tert-butyl)2-ethyl( R,E )-4-((dimethylamino)methylene)-5-oxopyrrolidine-1,2-dicarboxylic acid Preparation of ester (B1-4)

Dissolve 1-(tert-butyl)2-ethyl( R )-5-oxopyrrolidine-1,2-dicarboxylate (13 g, 50.5 mmol) in ethylene glycol dimethyl ether (70 mL ), then add 1-tert-butoxy- N,N,N',N' -tetramethylmethanediamine (14.09 g, 81 mmol), and the reaction solution is refluxed at 100°C for 10 hours. After the reaction is completed, the reaction solution is cooled to room temperature and then concentrated in vacuo to obtain crude product 1-(tert-butyl)2-ethyl( R,E )-4-((dimethylamino)methylene)- 5-Oxopyrrolidine-1,2-dicarboxylate (13.0 g, yield 82%), the crude product was used directly in the next step.

Step 3: Preparation of 1-(tert-butyl)2-ethyl( R )-4-methylene-5-oxopyrrolidine-1,2-dicarboxylate (B1-5)

1-(tert-butyl)2-ethyl( R,E )-4-((dimethylamino)methylene)-5-oxopyrrolidine-1,2-dicarboxylate (13.0 g, 50.5 mmol) was dissolved in tetrahydrofuran (80 mL), then 1N hydrochloric acid (25 mL) was added, and the reaction solution was stirred at 25°C for 2 hours. After the reaction is completed, wait for the reaction solution to separate naturally, extract the aqueous phase twice with tetrahydrofuran (20 mL), combine the organic phases, and then add potassium carbonate (10.13 g, 73.3 mmol) and 37% formaldehyde solution ( 40 mL), the reaction solution was stirred at 25°C for 45 min. After the reaction is completed, concentrate the reaction solution, add water (20 mL) to dissolve, and extract with ethyl acetate (40 mL*3). Combine the organic layers, wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, and concentrate to obtain the crude product. . The crude product was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 10:1 to 2:1) to obtain 1-(tert-butyl)2-ethyl(R)-4-methylene-5. -Oxopyrrolidine-1,2-dicarboxylate (6.0 g, yield 44.1%).

LC-MS, M/Z (ESI): 170.0 [M-100] ⁺ .

¹ H NMR (400 MHz, CDCl ₃ ) δ 6.23 (t, J = 2.8 Hz, 1H), 5.51 (t, J = 2.4 Hz, 1H), 4.60 (dt, J = 8.8, 4.4 Hz, 1H), 4.27 – 4.16 (m, 2H), 3.07 (ddt, J = 17.3, 10.1, 3.0 Hz, 1H), 2.71 (ddd, J = 17.5, 5.4, 2.4 Hz, 1H), 1.51 (s, 9H), 1.31 – 1.23 (m, 3H).

Step 4: 5-(tert-butyl)6-ethyl( R )-4-oxo-5-azaspiro[2.4]heptane-5,6-dicarboxylate (B1-6) Preparation

1-(tert-Butyl)2-ethyl( R )-4-methylene-5-oxopyrrolidine-1,2-dicarboxylate (6.0 g, 134 mmol) and palladium acetate (0.5 g, 2.228 mmol) was dissolved in diethyl ether (10 mL), then replaced with nitrogen three times, and vacuum ester was applied to the rubber stopper to prevent air leakage. Lower the reaction solution to -10°C, slowly add newly prepared diazomethane ether solution (5.62 g, 134 mmol), and stir the reaction solution at -10°C for 0.5 hours. Then the temperature of the reaction solution was raised to 25°C, and the reaction solution was continued to stir at 25°C for 10 hours. After the reaction is completed, the reaction solution is filtered through celite, then rinsed with ethyl acetate, and concentrated to obtain 5-(tert-butyl)6-ethyl( R )-4-oxo-5-azaspiro[2.4 ]Heptane-5,6-dicarboxylate (6.0 g, yield 95%).

LC-MS, M/Z (ESI): 184.3 [M-100] ⁺ .

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

Dissolve 5-(tert-butyl)6-ethyl( R )-4-oxo-5-azaspiro[2.4]heptane-5,6-dicarboxylate (6.0 g, 21.18 mmol) in To dichloromethane (60.0 mL), add hydrochloric acid/ethyl acetate (20 mL, 4M) at 0°C, then heat the reaction solution to 25°C, and stir the reaction solution at 25°C for 2 hours. After the reaction was completed, the reaction solution was directly spun to dryness and concentrated to obtain 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] ⁺ .

Step 6: Preparation of: ( R )-4-oxo-5-azaspiro[2.4]heptane-6-carboxylic acid (B1-8)

Dissolve ( R )-4-oxo-5-azaspiro[2.4]heptane-6-carboxylic acid ethyl ester (4.7 g, 25.7 mmol) in tetrahydrofuran (27 mL), water (18 mL) and methanol (3 mL) solution, and then add lithium hydroxide monohydrate (3.23 g, 77 mmol), and the reaction solution was stirred at 25°C for 16 hours. After the reaction is completed, the reaction solution is concentrated, and then extracted with ethyl acetate (20 mL). The impurities are extracted into the organic phase. Then the pH value of the aqueous phase is adjusted to about 3 with 1N hydrochloric acid, and then dichloromethane (30 mL) is used. mL*5), combine the organic phases, dry over anhydrous sodium sulfate, filter and concentrate to obtain 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 (400 MHz, CDCl ₃ ) δ 10.19 (s, 1H), 7.65 (s, 1H), 4.13 (q, J = 7.1 Hz, 1H), 2.08 (d, J = 19.5 Hz, 2H), 1.26 (t, J = 7.1 Hz, 2H), 1.21 – 1.15 (m, 1H), 0.91 – 0.79 (m, 1H).

Step 7: ( R )- N -(5-chloro-2,4-difluorophenyl)-4-oxo-5-azaspiro[2.4]heptane-6-formamide (B1-10 ) preparation

Combine ( 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) Dissolve in N,N-dimethylformamide (25 mL), then add 1-propylphosphonic anhydride (T ₃ P) in ethyl acetate (24.61 g, 38.7 mmol, 50% ethyl acetate) and Pyridine (6.26 mL, 77 mmol), the reaction solution was heated to 50°C and stirred under this condition for 10 hours. After the reaction is completed, add water (10 mL) and ethyl acetate (20 mL) to the reaction solution to dissolve, separate the organic phase, extract the aqueous phase with ethyl acetate (20 mL*3), combine the organic phases, and use brine ( 20 mL), washed and dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 10:1 to 5:1) to obtain ( R ) -N- (5-chloro-2,4-difluorophenyl)-4- Oxo-5-azaspiro[2.4]heptane-6-methamide (3.6 g, yield 77%).

LC-MS, M/Z (ESI): 301.6 [M+H] ⁺ .

Step 8: ( R ) -N- (5-chloro-2,4-difluorophenyl)-5-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-4- Preparation of oxo-5-azaspiro[2.4]heptane-6-methamide (B1-12)

( R )- N -(5-chloro-2,4-difluorophenyl)-4-oxo-5-azaspiro[2.4]heptane-6-formamide (1.6 g, 5.32 mmol) Dissolve 2-chloro-6-methyl-4-(trifluoromethyl)pyridine (1.249 g, 6,39 mmol) in 1,4-dioxane (15 mL), then add potassium carbonate (2.206 g, 15.96 mmol), 4,5-bisdiphenylphosphine-9,9-dimethylxanthene (XantPhos, 0.616 g, 1.064 mmol), and tris(dibenzylideneacetone)dipalladium (Pd ₂ dba ₃ , 0.487 g, 6.39 mmol), then replaced with nitrogen three times, and reacted at 100°C for 16 hours. After the reaction is completed, cool the reaction solution to room temperature, add water (5 mL) and ethyl acetate (10 mL) to the reaction solution, separate the organic phase, and extract the aqueous phase with ethyl acetate (10 mL*3). The organic phases were combined, washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain crude product. The crude product was purified by column chromatography (silica gel, ethyl acetate:petroleum ether = 8:1 to 3:1) to obtain ( R ) -N- (5-chloro-2,4-difluorophenyl)-5- (6-Methyl-4-(trifluoromethyl)pyridin-2-yl)-4-oxo-5-azaspiro[2.4]heptane-6-methamide (517 mg, yield 21.13% ).

LC-MS, M/Z (ESI): 460.2 [M+H] ⁺ .

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.94 (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, 3.9 Hz, 1H), 1.37 – 1.30 (m, 1H) , 1.20 – 1.13 (m, 1H), 0.99 – 0.93 (m, 1H).

Step 9: ( R ) -N- (5-chloro-2,4-difluorophenyl) -N -methyl-5-(6-methyl-4-(trifluoromethyl)pyridine-2- base)-4-oxo-5-azaspiro[2.4]heptane-6-methamide ( 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-formamide (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 then added, the reaction was heated to 50°C, and the reaction was stirred at 50°C for 1 hour. After the reaction is completed, cool the reaction solution to room temperature, add water (3 mL) and ethyl acetate (5 mL) to the reaction solution, separate the organic phase, and extract the aqueous phase with ethyl acetate (5 mL*3). The organic phases were combined, washed with brine (5 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain crude product. The crude product was separated and purified by reversed-phase high performance liquid chromatography to obtain ( 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-methamide (357 mg, 67% yield).

LC-MS, M/Z (ESI): 473.8 [M+H] ⁺ .

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.64 – 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).

The preparation of compound I-1B can be obtained by referring to the preparation method of I-1A. Compound number Structural formula name LC-MS, M/Z (ESI) I-1B (S)-N-(5-chloro-2,4-difluorophenyl)-N-methyl-5-(6-methyl-4-trifluoromethyl)pyridin-2-yl)-4- Oxo-5-azaspiro[2.4]heptane-6-formamide 473.8

Example 2 Preparation of Compound I-4

The synthesis 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-dicarboxylate (2.0 g, 7.54 mmol) in methanol (20 mL) at room temperature ) solution was added water (2 mL). Lithium hydroxide monohydrate (LiOH ^. H ₂ O, 92.2 g, 365.4 mmol) was added to the above reaction solution under stirring, and the reaction solution was stirred at 25°C for 12 h. After the reaction is complete, the reaction solution is concentrated to dryness, 10 mL of water is added, and the pH is adjusted to 3 with 1M hydrochloric acid aqueous solution while stirring. The reaction mixture was extracted with ethyl acetate (20 mL*3), the organic phases were combined, dried over sodium sulfate, and concentrated to obtain crude product ( S )-1-(tert-butoxycarbonyl)-4,4-difluoropyrrolidine-2 -Formic acid (1.4 g, 74% yield).

LC-MS, M/Z (ESI): 252.0 [M+H] ⁺ .

Second step: ( S )-2-((5-chloro-2,4-difluorophenyl)aminoformyl)-4,4-difluoropyrrolidine-1-carboxylic acid tert-butyl ester (B4 -Synthesis of 3)

Add ( S )-1-(tert-butoxycarbonyl)-4,4-difluoropyrrolidine-2-carboxylic acid (1.4 g, 5.6 mmol), 5-chloro-2,4 to a 100 mL round-bottomed flask. -Difluoroaniline (1.09 g, 6.7 mmol) and N,N-dimethylformamide (15 mL), cool the reaction solution to 0-5°C, add N,N-diisopropylethylamine to the reaction (2.16 g, 16.7 mmol), then slowly add 1-propylphosphoric acid cyclic anhydride (T ₃ P, 4.25 g, 6.68 mmol, 50% N,N-dimethylformamide solution), and the reaction rises to room temperature Stir for 2h. Add saturated sodium bicarbonate solution (40 mL) to the reaction to quench the reaction, then add 50 mL ethyl acetate, separate and collect the organic phase, wash the organic phase with saturated sodium bicarbonate solution (40 mL*2), and then Wash with saturated NaCl (40 mL) solution, dry over sodium sulfate, and concentrate to obtain a crude product. The crude product is purified with a silica gel column (ethyl acetate/petroleum ether = 1/5) to obtain ( S )-2-((5-chloro-2 , 4-difluorophenyl)aminoformyl)-4,4-difluoropyrrolidine-1-carboxylic acid tert-butyl ester (1.7 g, yield 77%).

LC-MS, M/Z (ESI): 397.1 [M+H] ⁺ .

The third step: ( S )-2-((5-chloro-2,4-difluorophenyl)(methyl)aminoformyl)-4,4-difluoropyrrolidine-1-carboxylic acid third step Synthesis of butyl ester (B4-4)

( S )-2-((5-chloro-2,4-difluorophenyl)aminoformyl)-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 0.5 h, methyl iodide (MeI, 0.9 g, 6.43 mmol), the reaction solution continued to stir at room temperature for 2 h. Add water (50 mL) and ethyl acetate (50 mL) to the reaction solution, separate and collect the organic phase, wash the organic phase with water (50 mL*2), dry over sodium sulfate, and concentrate to obtain crude product ( S )-2-( (5-Chloro-2,4-difluorophenyl)(methyl)aminoformyl)-4,4-difluoropyrrolidine-1-carboxylic acid tert-butyl ester (1.6 g, yield 88%) , used directly in the next step.

LC-MS, M/Z (ESI): 411.1 [M+H] ⁺ .

Step 4: ( S )-5-((5-chloro-2,4-difluorophenyl)(methyl)aminoformyl)-3,3-difluoro-2-oxopyrrolidine- Synthesis of 1-tert-butylcarboxylate (B4-5)

Add sodium periodate (4.03 g, 18.8 mmol) and water (45 mL) to the three-necked flask, replace the reaction system with nitrogen three times, and add ruthenium trichloride (RuCl ₃ , 0.78 g, 3.77 mmol) to the reaction system. and stirred at room temperature for 0.5 h. Slowly add ( S )-2-((5-chloro-2,4-difluorophenyl)(methyl)aminoformyl)-4,4-difluoropyrrolidine-1- After the addition of a solution of tert-butyl formate (1.6 g, 3.77 mmol) in ethyl acetate (15 mL), the reaction solution continued to stir for 16 h. Add isopropyl alcohol (5 mL) to the reaction solution and continue stirring at room temperature for 3 h. Add ethyl acetate (50 mL), extract the aqueous phase with ethyl acetate (10 mL*2), combine the organic phases, and concentrate. The crude product was obtained, and the crude product was purified with a silica gel column (ethyl acetate/petroleum ether=1/5) to obtain ( S )-5-((5-chloro-2,4-difluorophenyl)(methyl)aminoformamide (0.15 g, yield 9.4%).

LC-MS, M/Z (ESI): 425.1 [M+H] ⁺ .

The fifth step: ( S ) -N- (5-chloro-2,4-difluorophenyl)-4,4-difluoro- N -methyl-5-oxopyrrolidine-2-methamide ( Synthesis of B4-6)

( S )-5-((5-chloro-2,4-difluorophenyl)(methyl)aminoformyl)-3,3-difluoro-2-oxopyrrolidine-1-carboxylic acid The third butyl ester (0.15 g, 0.35 mmol) was dissolved in dichloromethane (3 mL), zinc bromide (ZnBr ₂ , 0.32 g, 1.41 mmol) was added to the reaction solution, and the reaction solution was stirred at room temperature for 4 h. After the reaction is complete, add saturated sodium bicarbonate solution (5 mL) to the reaction system, collect the organic phase by liquid separation, and wash the organic phase with saturated sodium bicarbonate solution (5 mL) and water (5 mL). Dry, filter and concentrate to obtain crude product ( S ) -N- (5-chloro-2,4-difluorophenyl)-4,4-difluoro- N -methyl-5-oxopyrrolidine-2-methyl Amide (0.11 g, yield 96%) was used directly in the next step.

LC-MS, M/Z (ESI): 325.0 [M+H] ⁺ .

Step 6: ( S ) -N- (5-chloro-2,4-difluorophenyl)-4,4-difluoro- N -methyl-1-(6-methyl-4-(trifluoro Synthesis of methyl)pyridin-2-yl)-5-oxopyrrolidine-2-methamide (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-bisdiphenylphosphine-9,9-dimethyloxide Heteroanthracene (0.02 g, 0.034 mmol), 2-chloro-6-methyl-4-(trifluoromethyl)pyridine (0.1 g, 0.51 mmol) were dissolved in 1,4-dioxane (2 mL ) solution, the reaction system was replaced with argon three times, and then reacted at 100°C for 12 h. The reaction solution was filtered, and the filtrate was concentrated to dryness to obtain a crude product. The crude product was purified on a silica gel plate to obtain (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 (400 MHz, DMSO-d6) δ 8.41–8.34 (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 synthesis route is shown below:

Step 1: 1-(tert-butyl)2-methyl( S )-4-(((trifluoromethyl)sulfonyl)oxy)-2,5-dihydro- 1H -pyrrole- Synthesis of 1,2-dicarboxylate (B5-2)

Dissolve 1-(tert-butyl)2-methyl( S )-4-oxopyrrolidine-1,2-dicarboxylate (5.0 g, 20.54 mmol) in tetrahydrofuran (50 mL) under nitrogen protection. , the reaction solution was cooled to -78 ℃. Lithium bistrimethylsilylamide (24.6 mL, 24.6 mmol, 1 mol/L tetrahydrofuran solution) was slowly added to the reaction under stirring, and the reaction solution was stirred at -78 °C for 1 h. A solution of N-phenylbis(trifluoromethanesulfonyl)imine (9.6 g, 24.6 mmol) in tetrahydrofuran (10 mL) was slowly added, and the reaction solution continued to stir at -78 °C for 2 h. Add 50 mL of saturated sodium bicarbonate to the reaction solution to quench the reaction. The reaction solution rises to room temperature. Add 100 mL of ethyl acetate to separate the layers. Extract the aqueous phase with ethyl acetate (20 mL*3) and combine the organic phases. , wash the organic phase with saturated sodium chloride solution (50 mL), dry over anhydrous sodium sulfate, and concentrate to obtain a crude product. The crude product is purified with a silica gel column (ethyl acetate/petroleum ether = 1/10, V/V) to obtain 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%).

Step 2: 1-(tert-butyl)2-methyl( S )-4-methyl-2,5-dihydro- 1H -pyrrole-1,2-dicarboxylate (B5-3) Synthesis

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

LC-MS, M/Z (ESI): 242.1 [M+H] ⁺ .

Step 3: 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-1,2-dicarboxylate (0.4 g, 1.66 mmol) at room temperature ) was added to a solution of methanol solution (4 mL). Water (0.4 mL) was added. Lithium hydroxide monohydrate (LiOH ^. H ₂ O, 139 mg, 3.32 mmol) was added to the above reaction under stirring. The reaction solution was stirred at 25°C for 12 h. After the reaction was complete, the reaction solution was concentrated to dryness, and 4 mL water, and adjust pH=3 with 1M hydrochloric acid aqueous solution under stirring. The reaction mixture was extracted with ethyl acetate (5 mL*3), the organic phases were combined, dried over sodium sulfate, and concentrated to obtain crude product (S)-1-(tert-butoxycarbonyl)-4-methyl-2,5-di Hydro- 1H -pyrrole-2-carboxylic acid (0.32 g, 85% yield).

LC-MS, M/Z (ESI): 228.1 [M+H] ⁺ .

Step 4: ( S )-2-((5-chloro-2,4-difluorophenyl)aminoformyl)-4-methyl-2,5-dihydro-1 H -pyrrole-1 -Synthesis of tert-butyl formate (B5-5)

Add ( S )-1-(tert-butoxycarbonyl)-4-methyl-2,5-dihydro- 1H -pyrrole-2-carboxylic acid (0.32 g, 1.4 mmol) to a 50 mL round-bottomed flask. , 5-chloro-2,4-difluoroaniline (276 mg, 1.7 mmol) and acetonitrile (3 mL). Add N-methylimidazole (347 mg, 4.22 mmol) to the reaction, then add N,N,N',N' -tetramethylchloroformamidine hexafluorophosphate (477 mg, 1.7 mmol), and react in the chamber Stir warm for 2 hours. Add saturated sodium bicarbonate solution (10 mL) to the reaction to quench the reaction, then add 50 mL ethyl acetate, separate and collect the organic phase, wash the organic phase with saturated sodium bicarbonate solution (10 mL*2), and then Wash with saturated sodium chloride (NaCl, 10 mL) solution, dry over sodium sulfate, and concentrate to obtain a crude product. The crude product is purified on a silica gel column (ethyl acetate/petroleum ether = 1/5) to obtain ( S )-2-((5- Chloro-2,4-difluorophenyl)aminoformyl)-4-methyl-2,5-dihydro- 1H -pyrrole-1-carboxylic acid tert-butyl ester (0.42g, yield 82% ).

LC-MS, M/Z (ESI): 373.1 [M+H] ⁺ .

Step 5: (2 S )-2-((5-chloro-2,4-difluorophenyl)aminoformyl)-3,4-dihydroxy-4-methylpyrrolidine-1-carboxylic acid Synthesis of tert-butyl ester (B5-6)

Under a nitrogen atmosphere, combine potassium carbonate (467.1 mg, 3.38 mmol), potassium ferricyanide (1.18 mg, 3.38 mmol), methanesulfonamide (107 mg, 1.13 mmol), and potassium osmate dihydrate (17.5 mg, 0.056 mmol). ) and hydroquinine 1,4-(2,3-naphthyridine) diether (86 mg, 0.11 mmol) were added to water (3 mL) and tert-butanol (1.5 mL). The reaction solution was lowered to 0-5°C and stirred at 0-5°C for 0.5 h, and then (S)-2-((5-chloro-2,4-difluorophenyl)aminoformyl) was slowly added -A solution of tert-butyl 4-methyl-2,5-dihydro-1H-pyrrole-1-carboxylate (0.42 g, 1.13 mmol) in tert-butanol (1.5 mL), the reaction was continued at 0-5 °C Stir for 12 hours. Ethyl acetate (10 mL) was added to the reaction, and the organic phase was collected by liquid separation. The organic phase was washed with saturated sodium chloride (NaCl, 10 mL) solution, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product, which was purified by a silica gel column ( Ethyl acetate/petroleum ether=1/1) to obtain (2 S )-2-((5-chloro-2,4-difluorophenyl)aminoformyl)-3,4-dihydroxy-4- Methylpyrrolidine-1-carboxylic acid tert-butyl ester (0.24 g, yield 52%).

LC-MS, M/Z (ESI): 407.1 [M+H] ⁺ .

Step 6: (3a R, 6 S, 6a S )-6-((5-chloro-2,4-difluorophenyl)aminoformyl)-2,2,3a-trimethyldihydrogen Synthesis of -3aH-[1,3]dioxolane[4,5-c]pyrrole-5(4H)-carboxylic acid tert-butyl ester (B5-7)

Under a nitrogen atmosphere, (2 S )-2-((5-chloro-2,4-difluorophenyl)aminoformyl)-3,4-dihydroxy-4-methylpyrrolidine-1- Tert-butyl formate (0.24 g, 0.59 mmol), pyridine 4-methylbenzenesulfonate (37 mg, 0.15 mmol), and 2,2-dimethoxypropane (307 mg, 2.95 mmol) were added to toluene ( 3 mL). The reaction solution was heated to 100 °C and stirred for 2 h. The reaction solution was then concentrated to obtain a crude product, which was purified by a silica gel column (ethyl acetate/petroleum ether = 1/3) to obtain (3a R , 6 S , 6a S )-6-((5-chloro-2,4-difluoro) Phenyl)aminoformyl)-2,2,3a-trimethyldihydro-3aH-[1,3]dioxolane[4,5-c]pyrrole-5(4H)-carboxylic acid tertiary Butyl ester (0.17g, yield 64%).

LC-MS, M/Z (ESI): 447.1 [M+H] ⁺ .

Step 7: (3a R ,6 S ,6a S )-6-((5-chloro-2,4-difluorophenyl)(methyl)aminoformyl)-2,2,3a-tris Synthesis of methyldihydro-3aH-[1,3]dioxola[4,5-c]pyrrole-5(4H)-carboxylic acid tert-butyl ester (B5-8)

(3a R ,6 S ,6a S )-6-((5-chloro-2,4-difluorophenyl)aminoformyl)-2,2,3a-trimethyldihydro-3aH- [1,3]Dioxola[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 ), add cesium carbonate (186 mg, 0.57 mmol), and the reaction solution is stirred at room temperature for 0.5 h. Methyl iodide (MeI, 81 mg, 0.57 mmol) is added, and the reaction solution continues to stir at room temperature for 2 h. Water (5 mL) and ethyl acetate (5 mL) were added to the reaction solution, and the organic phase was collected by liquid separation. The organic phase was washed with water (5 mL*3), dried over sodium sulfate, and concentrated to obtain crude product (3a R , 6 S , 6a S )-6-((5-chloro-2,4-difluorophenyl)(methyl)aminoformyl)-2,2,3a-trimethyldihydro-3aH-[1,3 Dioxolane[4,5-c]pyrrole-5(4H)-carboxylic acid tert-butyl ester (160 mg, yield 91%) was used directly in the next step.

LC-MS, M/Z (ESI): 461.1 [M+H] ⁺ .

Step 8: (3a S ,6 S ,6a S )-6-((5-chloro-2,4-difluorophenyl(methyl)aminoformyl)-2,2,3a-trimethyl Synthesis of tert-butyl-4-oxodihydro-3aH-[1,3]dioxolane[4,5- c ]pyrrole-5(4H)-carboxylate (B5-9)

Add sodium periodate (223 mg, 1.04 mmol) and water (2 mL) to the three-necked flask, replace the reaction system with nitrogen three times, and add hydrated ruthenium dioxide (RuO ₂ ^. xH ₂ O, 5.2 mg, 0.035 mmol) and stirred at room temperature for 0.5 h. Slowly add (3a R ,6 S ,6a S )-6-((5-chloro-2,4-difluorophenyl)(methyl)aminoformyl)-2,2 into the reaction, 3a-Trimethyldihydro-3aH-[1,3]dioxola[4,5-c]pyrrole-5(4H)-carboxylic acid tert-butyl ester (0.16 g, 0.35 mmol) in ethyl acetate ( 2 mL) solution, after the addition, the reaction solution continued to stir for 16 h. Add isopropyl alcohol (1 mL) to the reaction solution and continue stirring at room temperature for 3 h. Add ethyl acetate (5 mL), extract the aqueous phase with ethyl acetate (5 mL*2), combine the organic phases, and concentrate. The crude product was obtained, and the crude product was purified by silica gel column (ethyl acetate/petroleum ether=1/3) to obtain (3a S , 6 S , 6a S )-6-((5-chloro-2,4-difluorophenyl(methyl) )Aminoformyl)-2,2,3a-trimethyl-4-oxodihydro-3aH-[1,3]dioxolane[4,5- c ]pyrrole-5(4H)- Tert-butyl formate (0.11 g, yield 67%).

LC-MS, M/Z (ESI): 475.1 [M+H] ⁺ .

Step 9: (3a S ,4 S ,6a S )-N-(5-chloro-2,4-difluorophenyl)-N,2,2,6a-tetramethyl-6-oxotetrahydro Synthesis of -3aH-[1,3]dioxola[4,5-c]pyrrole-4-methamide (B5-10)

(3a S ,6 S ,6a S )-6-((5-chloro-2,4-difluorophenyl(methyl)aminoformyl)-2,2,3a-trimethyl-4 -Oxodihydro-3aH-[1,3]dioxola[4,5- c ]pyrrole-5(4H)-carboxylic acid tert-butyl ester (0.11 g, 0.23 mmol) was dissolved in dichloromethane (2 mL) solution, add zinc bromide (ZnBr ₂ , 209 mg, 0.93 mmol) to the reaction solution, and stir the reaction solution at room temperature for 4 h. LC-MS shows that the reaction is complete, and saturated hydrogen carbonate is added to the reaction system. Sodium solution (5 mL), separate and collect the organic phase, wash the organic phase with saturated sodium bicarbonate solution (5 mL) and water (5 mL), dry over sodium sulfate, filter, and concentrate to obtain crude products (3a S , 4 S ,6a S )- N -(5-chloro-2,4-difluorophenyl)- N ,2,2,6a-tetramethyl-6-oxotetrahydro-3aH-[1,3]dioxy Pentyl[4,5- c ]pyrrole-4-methamide (90 mg, yield 100%) was used directly in the next step.

LC-MS, M/Z (ESI): 375.1 [M+H] ⁺ .

Step 10: (3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-N,2,2,6a-tetramethyl-5-(6-methyl-4 -(Trifluoromethyl)pyridin-2-yl)-6-oxotetrahydro-3aH-[1,3]dioxolane[4,5-c]pyrrole-4-methamide (B5-11 )Synthesis

(3a S ,4 S ,6a S )-N-(5-chloro-2,4-difluorophenyl) -N ,2,2,6a-tetramethyl-6-oxotetrahydro-3a H -[1,3]dioxola[4,5- c ]pyrrole-4-methamide (90 mg, 0.24 mmol), potassium carbonate (68 mg, 0.48 mmol), tris(dibenzylideneacetone) Dipalladium (22 mg, 0.024 mmol), 4,5-bisdiphenylphosphine-9,9-dimethylxanthene (14 mg, 0.024 mmol), 2-chloro-6-methyl-4-( Trifluoromethyl)pyridine (70 mg, 0.36 mmol) was dissolved in 1,4-dioxane (2 mL) solution, the reaction system was replaced with argon three times, and then reacted at 100 °C for 12 h. The reaction solution was filtered, and the filtrate was concentrated to dryness to obtain a crude product, which was purified by a silica gel column (ethyl acetate/petroleum ether = 1/3) to obtain (3a S , 4 S , 6a S )-N-(5-chloro-2,4- Difluorophenyl) -N ,2,2,6a-tetramethyl-5-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-6-oxotetrahydro-3a H -[1,3]dioxola[4,5- c ]pyrrole-4-methamide (100 mg, yield 78%).

LC-MS, M/Z (ESI): 534.1 [M+H] ⁺ .

Step 11: ( 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-methamide (I-5 )

Under nitrogen protection, (3a S , 4 S , 6a S ) -N- (5-chloro-2,4-difluorophenyl) -N ,2,2,6a-tetramethyl-5-(6- Methyl-4-(trifluoromethyl)pyridin-2-yl)-6-oxotetrahydro-3a H -[1,3]dioxola[4,5-c]pyrrole-4-methane The amine (0.1 g, 0.187 mmol) was dissolved in a solution of dichloromethane (2 mL), and a solution of boron trichloride (BCl ₃ ) in dichloromethane (0.38 mL, 0.38 mmol, 1 mol/L) was added. The reaction solution was stirred at room temperature for 2 h, saturated sodium bicarbonate solution (2 mL) was added to quench the reaction, 5 mL of methylene chloride was added, and the organic phase was collected by separation. The organic phase was separated with saturated sodium bicarbonate solution (5 mL). ) and water (5 mL), dried over anhydrous sodium sulfate, and concentrated to obtain a crude product, which was purified by silica gel plate (ethyl acetate/petroleum ether) to obtain (2 S , 3 S , 4 S ) -N- (5-chloro- 2,4-Difluorophenyl)-3,4-dihydroxy- N ,4-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5- Oxopyrrolidine-2-methamide (I-5 ) (36 mg, yield 39%).

¹ H NMR (400 MHz, Methanol-d4) δ 8.57–8.39 (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 synthesis route is shown below:

Step 1: (2S)-1-(tert-butoxycarbonyl)-4-hydroxy-4-methylpyrrolidine-2-carboxylic acid (B10-2) (2S)-1-(tert-butoxycarbonyl)-4 -hydroxy-4-methylpyrrolidine-2-carboxylic acid

Under nitrogen protection, dissolve tert-butyl (5-chloro-2,4-difluorophenyl)aminomethyl ester (B10-1, 10.0 g, 43.6 mmol) in tetrahydrofuran (110 mL), and then Methyl magnesium bromide (3 M, 43.62 mL) was slowly added at -20°C, and the reaction solution was stirred at 25°C for 1 hour. After the reaction is completed, slowly drop 200 mL of saturated hydrochloric acid (1M) solution at 0°C to quench, adjust the pH value to 3~4, then extract with dichloromethane (200 mL*3), and use saturated chlorine to quench the organic phase. Wash with sodium chloride solution (200 mL), dry with anhydrous sodium sulfate, filter and concentrate to obtain (2S)-1-(tert-butoxycarbonyl)-4-hydroxy-4-methylpyrrolidine-2-carboxylic acid (B10-2 , 11.0 g, crude product).

Step 2: 2-Benzyl 1-(tert-butyl) (2S,4S)-4-hydroxy-4-methylpyrrolidine-1,2-dicarboxylate (B10-3) 2-benzyl 1-(tert-butyl) (2S,4S)-4-hydroxy-4-methylpyrrolidine-1,2-dicarboxylate

Under nitrogen protection, (2S)-1-(tert-butoxycarbonyl)-4-hydroxy-4-methylpyrrolidine-2-carboxylic acid (B10-2, 10.0 g, 43.6 mmol) was dissolved in N,N -dimethylformamide (110 mL), then slowly add cesium carbonate (3eq) and benzyl bromide (4eq) at 0°C, and stir the reaction solution at 25°C for 12 hours. After the reaction is completed, add water (300 mL) for quenching, then extract with ethyl acetate (100 mL*3), wash the organic phase with saturated sodium chloride solution (800 mL), dry over anhydrous sodium sulfate, filter and concentrate to obtain The crude product was separated and purified by reversed-phase high performance liquid chromatography to obtain 2-benzyl 1-(tert-butyl) (2S,4S)-4-hydroxy-4-methylpyrrolidine-1,2-di Formate (B10-3, 4.00 g, yield 26.6%).

LCMS, M/Z (ESI): 236.1[M+H] ⁺ .

Step 3: (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-dicarboxylate (B10-3, 1.00 g, 288 mmol) and palladium hydroxide/carbon (10%Wt, 0.60 g) were dissolved in ethanol (30.0 mL), after three replacements with nitrogen and three replacements with hydrogen. The reaction solution was stirred under hydrogen gas (50 Psi) pressure at 25°C for 12 hours. After the reaction is completed, the reaction solution is filtered and concentrated to obtain crude (2S, 4S)-1-(tert-butoxycarbonyl)-4-hydroxy-4-methylpyrrolidine-2-carboxylic acid (B10-4, 1.00 g). used directly in the next step.

Step 4: tertiary 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

Combine (2S,4S)-1-(tert-butoxycarbonyl)-4-hydroxy-4-methylpyrrolidine-2-carboxylic acid (B10-4, 400 mg, 1.63 mmol) and 5-chloro-2, 4-Difluoroaniline (240 mg, 1.47 mmol) was dissolved in acetonitrile (5.00 mL), and N-methylimidazole (401 mg, 4.89 mmol) and N,N,N,N-tetramethylchloroformamidine were added successively. Hexafluorophosphoric acid (732 mg, 2.61 mmol), the reaction solution was stirred at 80°C for 12 hours. After the reaction is completed, the reaction solution is directly concentrated under reduced pressure to obtain a crude product, which is separated by column chromatography (the mobile phase is petroleum ether: ethyl acetate = 20:1 - 5:1) to obtain tert-butyl (2S, 4S)-2 -((5-chloro-2,4-difluorophenyl)aminocarbonyl)-4-hydroxy-4-methylpyrrolidine-1-carboxylate (B10-5, 200 mg, yield 31.4%) .

LCMS, M/Z (ESI): 291.0[M+H] ⁺ .

Step 5: tertiary butyl(2S,4S)-2-((5-chloro-2,4-difluorophenyl)(methyl)aminocarbonyl)-4-hydroxy-4-methylpyrrolidine -1-Formyl 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, 100 mg, 255 μmol) and cesium carbonate (166 mg, 511 μmol) were dissolved in N,N-dimethylformamide (1.00 mL). After the reaction solution was stirred at 25°C for 30 minutes, methyl iodide ( 145 mg, 1.02 mmol). The reaction solution was stirred at 25°C for 2 hours. After the reaction was completed, the reaction was quenched with water (10 mL), then extracted with ethyl acetate (10.0 mL*3), the organic phase was washed with saturated sodium chloride solution (20.0 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to obtain the third Tributyl(2S,4S)-2-((5-chloro-2,4-difluorophenyl)(methyl)aminocarbonyl)-4-hydroxy-4-methylpyrrolidine-1-carboxylic acid Ester (B10-6, 150 mg, crude).

LCMS, M/Z (ESI): 427.3[M+Na] ⁺ .

Step 6: 3-Butyl(3S,5S)-5-((5-chloro-2,4-difluorophenyl)(methyl)aminocarbonyl)-3-hydroxy-3-methyl-2 -Oxylidenepyrrolidine-1-carboxylate (B10-7) tert-butyl(3S,5S)-5-((5-chloro-2,4-difluorophenyl)(methyl)carbamoyl)-3-hydroxy- 3-methyl-2-oxopyrrolidine-1-carboxylate

Under nitrogen protection, hydrated ruthenium trichloride (48.4 mg, 185 μmol) and sodium periodate (237 mg, 1.11 mmol) were dissolved in water (0.5 mL), replaced with nitrogen three times, and the reaction solution was stirred at 25°C. 30 min, then tert-butyl(2S,4S)-2-((5-chloro-2,4-difluorophenyl)(methyl)aminocarbonyl) dissolved in ethyl acetate (0.5 mL) -4-Hydroxy-4-methylpyrrolidine-1-carboxylate (B10-6, 150 mg, 370. μmol) was slowly added to the reaction solution, and finally, the reaction solution was stirred at 25°C for 12 hours. After the reaction is completed, add saturated sodium sulfite (10.0 mL) to quench the reaction, and then extract with ethyl acetate (10 mL*3). The organic phase is washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrate to obtain tert-butyl(3S,5S)-5-((5-chloro-2,4-difluorophenyl)(methyl)aminocarbonyl)-3-hydroxy-3-methyl-2-oxo Yethylenepyrrolidine-1-carboxylate (B10-7, 50.0 mg, crude).

LC-MS, M/Z (ESI): 419.1[M+H] ⁺ .

Step 7: (2S,4S)-N-(5-chloro-2,4-difluorophenyl)-4-hydroxy-N,4-dimethyl-5-oxyylidenepyrrolidine-2-methyl Amide (B10-8) (2S,4S)-N-(5-chloro-2,4-difluorophenyl)-4-hydroxy-N,4-dimethyl-5-oxopyrrolidine-2-carboxamide

tert-Butyl(3S,5S)-5-((5-chloro-2,4-difluorophenyl)(methyl)aminocarbonyl)-3-hydroxy-3-methyl-2-oxo Pyrrolidine-1-carboxylate (B10-7, 50.0 mg, 119 μmol) was dissolved in dichloromethane (1.00 mL), then zinc bromide (134 mg, 596 μmol) was added, and the reaction solution was heated at 25°C Stir for 4 hours. After the reaction is completed, quench the reaction with saturated sodium bicarbonate (10 mL), then extract with dichloromethane (5 mL*3), wash the organic phase with saturated sodium chloride solution (5 mL), dry with anhydrous sodium sulfate, and filter Concentrate to obtain (2S,4S)-N-(5-chloro-2,4-difluorophenyl)-4-hydroxy-N,4-dimethyl-5-oxyylidenepyrrolidine-2-methamide (B10-8, 22.0 mg, crude).

LC-MS, M/Z (ESI): 319.2[M+H] ⁺ .

Step 8: (2S,4S)-N-(5-chloro-2,4-difluorophenyl)-4-hydroxy-N,4-dimethyl-1-(6-methyl-4-( Trifluoromethyl)pyridin-2-yl)-5-oxyylidenepyrrolidine-2-methamide (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-oxyylidenepyrrolidine-2-methamide ( B10-8, 20 mg, 62.7 μmol) and 2-chloro-6-methyl-4-(trifluoromethyl)pyridine (24.5 mg, 125 μmol) were dissolved in 1,4-dioxane (1.00 mL) , then add cesium carbonate (40.8 mg, 125.51 μmmol), tris(dibenzylideneacetone)dipalladium (8.62 mg, 9.41 μmol) and 4,5-bis(diphenylphosphorus) respectively at room temperature and under nitrogen protection. -9,9-dimethylxanthene (10.8 mg, 18.8 μmol), after nitrogen replacement three times, the reaction solution was stirred at 100°C for 12 hours. After the reaction is completed, filter and concentrate directly to obtain a brown oil. The crude product is separated and purified by reversed-phase high performance liquid chromatography (separation method: Waters Xbridge 150*25mm*5um; mobile phase: [water (NH ₄ HCO ₃ ) -acetonitrile]; gradient: 48%-78% B over 9 min), after purification, (2S,4S)-N-(5-chloro-2,4-difluorophenyl)-4-hydroxy-N,4 was obtained -Dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxyylidenepyrrolidine-2-methamide (I-10, 1.60 mg, product rate 2.64%).

¹ H NMR (400 MHz, 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 synthesis route is as follows:

The first step: 1-(tert-butyl) 2-methyl(S)-4-cyclopropyl-2,5-dihydro-1H-pyrrole-1,2-dicarboxylate (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)-dicarboxylate ( B11-1, 15.0 g, 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) and replaced with nitrogen. Three times, tetrakis(triphenylphosphorus)palladium (4.62 g, 4.00 mmol) was added under nitrogen protection, and the reaction solution was stirred at 100°C for 12 hours. After the reaction is completed, it is directly filtered and concentrated to obtain the crude product. The crude product was separated and purified by column chromatography (the mobile phase was petroleum ether: ethyl acetate = 10:1 to 3:1). After purification, 1-(tert-butyl) 2-methyl (S)-4-cyclo was obtained. Propyl-2,5-dihydro-1H-pyrrole-1,2-dicarboxylate (B11-2, 3.70 g, yield 37.6%).

¹ H NMR (400 MHz, CHCl ₃ - d ) δ 5.22 (d, J =5.38 Hz, 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).

Step 2: (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-dicarboxylate (B11-2, 3.70 g, 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 solution was stirred at 25°C for 12 hours. After the reaction is completed, add dilute hydrochloric acid (1M) to adjust the pH to 3~4, then extract with ethyl acetate (50 mL*3), wash the organic phase with saturated sodium chloride solution (50 mL) and dry with anhydrous sodium sulfate. , filtered and concentrated to obtain crude product (S)-1-(tert-butoxycarbonyl)-4-cyclopropyl-2,5-dihydro-1H-pyrrole-2-carboxylic acid (B11-3, 3.20 g, yield 91.3 %), the crude product was directly used in the next step.

Step 3: tertiary butyl (S)-2-((5-chloro-2,4-difluorophenyl)aminocarbonyl)-4-cyclopropyl-2,5-dihydro-1H-pyrrole -1-Formyl ester (B11-4) tert-butyl(S)-2-((5-chloro-2,4-difluorophenyl)carbamoyl)-4-cyclopropyl-2,5-dihydro-1H-pyrrole-1 -carboxylate

Combine (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) was dissolved in acetonitrile (30.0 mL), and N,N,N,N-tetramethylchloroformamidine hexafluorophosphate (3.54 g, 12.6 mmol) and N-methyl were added respectively. Base imidazole (2.59 g, 31.5 mmol), the reaction solution was stirred at 25°C for 12 hours. After the reaction is completed, it is directly filtered and concentrated to obtain the crude product. The crude product was separated and purified by column chromatography (the mobile phase was petroleum ether: ethyl acetate = 10:1 to 5:1) to obtain tert-butyl (S)-2-((5-chloro-2,4-difluorobenzene) (3.80 g, yield: 90.5%).

¹ H NMR (400 MHz, 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).

Step 4: tertiary butyl(2S)-2-((5-chloro-2,4-difluorophenyl)aminocarbonyl)-4-cyclopropyl-3,4-dihydroxypyrrolidine-1 -Formate (B11-5) tert-butyl(2S)-2-((5-chloro-2,4-difluorophenyl)carbamoyl)-4-cyclopropyl-3,4-dihydroxypyrrolidine-1-carboxylate

Combine methane sulfonamide (1.02 g, 7.02 mmol), potassium carbonate (2.91 g, 21.06 mmol), potassium ferricyanide (6.93 g, 21.06 mmol), and hydrogenated quinine 1,4-(2,3-diaza Naphthalene diether (546.88 mg, 702.05 μmol) and potassium osmate dihydrate (129.34 mg, 351.03 μmol) were dissolved in tert-butanol (5.00 mL) and water (5.00 mL). The reaction liquid was replaced with nitrogen three times. Stir for 30 minutes at 0°C, then dissolve tert-butyl(S)-2-((5-chloro-2,4-difluorophenyl)aminocarbonyl)- in tert-butanol (5.00 mL). 4-Cyclopropyl-2,5-dihydro-1H-pyrrole-1-carboxylate (B11-4, 2.8 g, 7.02 mmol) was slowly added to the reaction solution at 0°C, and the reaction solution was stirred at 0°C. 12 hours. After the reaction is completed, filter and extract with ethyl acetate (20 mL*3), wash the organic phase with saturated sodium chloride solution (40 mL) and dry over anhydrous sodium sulfate, then filter and concentrate to obtain the crude product. The crude product was separated and purified by column chromatography (petroleum ether: ethyl acetate = 10:1 - 2:1) to obtain tert-butyl (2S)-2-((5-chloro-2,4-difluorophenyl)amine (carbonyl)-4-cyclopropyl-3,4-dihydroxypyrrolidine-1-carboxylate (B11-5, 600 mg, yield 19.7%).

LC-MS, M/Z (ESI): 307.2(M+H) ⁺ .

Step 5: 3-Butyl(3aR,6S,6aS)-6-((5-chloro-2,4-difluorophenyl)aminocarbonyl)-3a-cyclopropyl-2,2-dimethyl 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 The ester (B11-5, 435 mg, 973 μmol) and 2,2-dimethoxypropane (506 mg, 4.87 mmol) were dissolved in toluene (4.00 mL), and then pyridine-p-toluenesulfonate (61.1 mg, 243 μmol), the reaction solution was stirred at 100°C for 2 hours. After the reaction is completed, it is directly concentrated to obtain the crude product. The crude product was separated and purified by column chromatography (the mobile phase was petroleum ether: ethyl acetate = 10:1 - 5:1) to obtain tert-butyl (3aR,6S,6aS)-6-((5-chloro-2,4 -Difluorophenyl)aminocarbonyl)-3a-cyclopropyl-2,2-dimethyltetrahydro-5H-[1,3]dioxalan[4,5-c]pyrrole-5- Formate (B11-6, 230 mg, yield 46.8%).

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

Step 6: 3-butyl(3aR,6S,6aS)-6-((5-chloro-2,4-difluorophenyl)(methyl)aminocarbonyl)-3a-cyclopropyl-2, 2-Dimethyltetrahydro-5H-[1,3]dioxola[4,5-c]pyrrole-5-carboxylate (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(3aR,6S,6aS)-6-((5-chloro-2,4-difluorophenyl)aminocarbonyl)-3a-cyclopropyl-2,2-dimethyltetrahydro -5H-[1,3]dioxola[4,5-c]pyrrole-5-carboxylate (200 mg, 422 μmol) and cesium carbonate (275 mg, 845 μmol) were dissolved in N,N- After stirring the reaction solution in dimethylformamide (1.00 mL) for 30 minutes, methyl iodide (120 mg, 845 μmol) was slowly added dropwise, and the reaction solution was stirred at 25°C for 2 hours. After the reaction was completed, water (10 mL) was added to dilute the reaction, and then extracted with ethyl acetate (10 mL*3). The organic phase was washed with saturated sodium chloride solution (10 mL) and dried over anhydrous sodium sulfate, then filtered and concentrated to obtain the third Tributyl(3aR,6S,6aS)-6-((5-chloro-2,4-difluorophenyl)(methyl)aminocarbonyl)-3a-cyclopropyl-2,2-dimethyl Tetrahydro-5H-[1,3]dioxalan[4,5-c]pyrrole-5-carboxylate (B11-7, 180 mg, crude product), the crude product was used directly in the next step.

LC-MS, M/Z (ESI): 387.1[M+H-100] ⁺ .

Step 7: 3-butyl(3aS,6S,6aS)-6-((5-chloro-2,4-difluorophenyl)(methyl)aminocarbonyl)-3a-cyclopropyl-2, 2-Dimethyl-4-oxyylidenetetrahydro-5H-[1,3]dioxola[4,5-c]pyrrole-5-carboxylate (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

Under nitrogen protection, hydrated ruthenium dioxide (13.6 mg, 90.3 μmol) and sodium periodate (289.91 mg, 1.36 mmol) were dissolved in water (1.50 mL), replaced with nitrogen three times, and the reaction solution was stirred at 25°C for 30 minutes, then tert-butyl(3aR,6S,6aS)-6-((5-chloro-2,4-difluorophenyl)(methyl)aminocarbonyl) dissolved in ethyl acetate (1.00 mL) )-3a-cyclopropyl-2,2-dimethyltetrahydro-5H-[1,3]dioxola[4,5-c]pyrrole-5-carboxylate (B11-7, 220 mg, 451.81 μmol) was slowly added to the reaction solution, and the reaction solution was stirred at 25°C for 12 hours. After the reaction is completed, add saturated sodium sulfite (20.0 mL) to quench the reaction, and then extract with ethyl acetate (10 mL*3). The organic phase is washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrate to obtain crude product. The crude product was separated and purified by column chromatography (mobile phase was petroleum ether: ethyl acetate = 5:1 to 1:1) to obtain tert-butyl (3aS, 6S, 6aS)-6-((5-chloro-2,4 -Difluorophenyl)(methyl)aminocarbonyl)-3a-cyclopropyl-2,2-dimethyl-4-oxyylidenetetrahydro-5H-[1,3]dioxola[ 4,5-c]pyrrole-5-carboxylate (B11-8, 105 mg, yield 46.4%).

Step 8: (3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-6a-cyclopropyl-N,2,2-trimethyl-6-oxyylidene Tetrahydro-4H-[1,3]dioxola[4,5-c]pyrrole-4-methamide (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(3aS,6S,6aS)-6-((5-chloro-2,4-difluorophenyl)(methyl)aminocarbonyl)-3a-cyclopropyl-2,2-di Methyl-4-oxyylidenetetrahydro-5H-[1,3]dioxola[4,5-c]pyrrole-5-carboxylate (B11-8, 50.0 mg, 99.8 μmol) was dissolved in Dichloromethane (1.00 mL) was then added with zinc bromide (89.9 mg, 399 μmol), and the reaction solution was stirred at 25°C for 12 hours. After the reaction is completed, quench the reaction with saturated sodium bicarbonate (10 mL), then extract with dichloromethane (10 mL*3), wash the organic phase with saturated sodium chloride solution (10 mL), dry with anhydrous sodium sulfate, and filter Concentration gave the crude product, (3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-6a-cyclopropyl-N,2,2-trimethyl-6-oxyylidene Tetrahydro-4H-[1,3]dioxola[4,5-c]pyrrole-4-methamide (B11-9, 35.0 mg, yield 87.5%).

Step 9: (3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-6a-cyclopropyl-N,2,2-trimethyl-5-(6- Methyl-4-(trifluoromethyl)pyridin-2-yl)-6-oxyylidenetetrahydro-4H-[1,3]dioxola[4,5-c]pyrrole-4-methyl Amide (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

(3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-6a-cyclopropyl-N,2,2-trimethyl-6-oxyylidenetetrahydro- 4H-[1,3]dioxalan[4,5-c]pyrrole-4-carboxamide (B11-9, 50.0 mg, 124 μmol) and 2-chloro-6-methyl-4-( Trifluoromethyl)pyridine (36.6 mg, 18 μmol) was dissolved in 1,4-dioxane (2.00 mL), and then potassium carbonate (51.7 mg, 374 μmol) and tris(dioxane) were added under nitrogen protection. Benzyl acetone) dipalladium (10.7 mg, 18.7 μmol) and 4,5-bis(diphenylphosphorus)-9,9-dimethylxanthene (21.6 mg, 37.4 μmol), after three nitrogen replacements, the reaction The solution was stirred at 100°C for 3 hours. After the reaction is completed, the crude product is directly filtered and concentrated to obtain a brown oily substance. The crude product is separated and purified by column chromatography (mobile phase is petroleum ether: ethyl acetate = 3:1). After purification, (3aS, 4S, 6aS) is obtained. -N-(5-chloro-2,4-difluorophenyl)-6a-cyclopropyl-N,2,2-trimethyl-5-(6-methyl-4-(trifluoromethyl) Pyridin-2-yl)-6-oxyylidenetetrahydro-4H-[1,3]dioxola[4,5-c]pyrrole-4-methamide (B11-10, 60.0 mg, product rate 85.9%).

LC-MS, M/Z (ESI): 560.0[M+H] ⁺ .

Step 17: (2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-4cyclopropyl-3,4-dihydroxy-N-methyl-1-( 6-Methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxyylidenepyrrolidine-2-methamide (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

(3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-6a-cyclopropyl-N,2,2-trimethyl-5-(6-methyl- 4-(Trifluoromethyl)pyridin-2-yl)-6-oxyylidenetetrahydro-4H-[1,3]dioxola[4,5-c]pyrrole-4-methamide ( B11-10, 20 mg, 35.72 μmol) was dissolved in dichloromethane (1.00 mL), then a solution of boron trichloride in dichloromethane (1 M, 71.4 μL) was added at -20°C, and the reaction solution was heated at 25°C. Stir for 2 hours. After the reaction is completed, adjust the pH to 9 with saturated sodium bicarbonate solution, then extract with dichloromethane (10 mL*3), wash the organic phase with saturated sodium chloride solution (10.0 mL), dry with anhydrous sodium sulfate, filter and concentrate. Get crude product. The crude product was separated and purified by reversed-phase high performance liquid chromatography. The separation method was: Waters Xbridge C18 150*50mm*10um; mobile phase: [water (NH ₄ HCO ₃ )-acetonitrile]; gradient: 50%-80% B over 10 min, after purification, (2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-4cyclopropyl-3,4-dihydroxy-N-methyl-1- (6-Methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxyylidenepyrrolidine-2-methamide (I-11, 12.0 mg, yield 20.9%).

¹ H NMR (400 MHz, CH ₃ OH- d ₄ ) δ 8.32 - 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, 1 H), 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 synthesis route is as follows:

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

2-chloro-4-cyclopropyl-6-methylpyridine

Dissolve 2-chloro-4-iodo-6-methylpyridine (501 mg, 1.98 mmol) and cyclopropylboronic acid (220.72 mg, 2.57 mmol) in 1,4-dioxane (5.00 mL) and Potassium phosphate (1.26 g, 5.93 mmol), palladium acetate (66.56 mg, 296.49 μmol) and tricyclohexylphosphine (166.29 mg, 592.98 μmol) were added under nitrogen protection. After nitrogen replacement three times, the reaction solution was stirred at 100°C. 3 hours. After the reaction is completed, the crude product is directly filtered and concentrated to obtain a brown oily substance. The crude product is separated and purified by column chromatography (the mobile phase is petroleum ether: ethyl acetate = 15:1). After purification, 2-chloro-4-cyclohexane is obtained. Propyl-6-methylpyridine (150 mg, yield 45.3%).

¹ H NMR (400 MHz, 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).

Step 2: (3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-5-(4-cyclopropyl-6-methylpyridin-2-yl)-2 ,2,6a-trimethyl-N-(methyl)-6-oxyylidenetetrahydro-4H-[1,3]dioxola[4,5-c]pyrrole-4-methamide (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

(3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-2,2,6a-trimethyl-N-(methyl)-6-oxyylidenetetrahydrogen -4H-[1,3]dioxalan[4,5-c]pyrrole-4-carboxamide (B5-10, 150 mg, 400.25 μmol) and 2-chloro-4-cyclopropyl-6 -Picoline (134.19 mg, 800.50 μmol) was dissolved in 1,4-dioxane (2.00 mL), and then cesium carbonate (260.82 mg, 800.50 μmol) and tris(dibenzylidene) were added under nitrogen protection. Acetone) dipalladium (54.98 mg, 60.04 μmol) and 4,5-bis(diphenylphosphorus)-9,9-dimethylxanthene (69.48 mg, 120.07 μmol). After nitrogen replacement three times, the reaction solution was Stir at 100°C for 3 hours. After the reaction is completed, the crude product is directly filtered and concentrated to obtain a brown oily substance. The crude product is separated and purified by column chromatography (mobile phase is petroleum ether: ethyl acetate = 3:1). After purification, (3aS, 4S, 6aS) is obtained. -N-(5-chloro-2,4-difluorophenyl)-5-(4-cyclopropyl-6-methylpyridin-2-yl)-2,2,6a-trimethyl-N- (Methyl)-6-oxyylidenetetrahydro-4H-[1,3]dioxola[4,5-c]pyrrole-4-methamide (B20-3, 20.0 mg, crude).

LC-MS, M/Z (ESI): 506.1[M+H] ⁺ .

Step 3: (2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-1-(4-cyclopropyl-6-methylpyridin-2-yl)-3 ,4-dihydroxy-4-methyl-N-(methyl)-5-oxyylidenepyrrolidine-2-methamide ( 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,6aS)-N-(5-chloro-2,4-difluorophenyl)-5-(4-cyclopropyl-6-methylpyridin-2-yl)-2,2, 6a-Trimethyl-N-(methyl)-6-oxyylidenetetrahydro-4H-[1,3]dioxalan[4,5-c]pyrrole-4-methamide (B20- 3, 10 mg, 19.77 μmol) was dissolved in dichloromethane (1.00 mL), then a dichloromethane solution of boron trichloride (1 M, 39.5 L) was added at -20°C, and the reaction solution was stirred at 25°C. 1 hour. After the reaction is completed, adjust the pH to 9 with saturated sodium bicarbonate solution, then extract with dichloromethane (10 mL*2), wash the organic phase with saturated sodium chloride solution (10.0 mL), dry with anhydrous sodium sulfate, filter and concentrate. Get crude product. The crude product was separated and purified by reversed-phase high performance liquid chromatography. The separation method was: Waters Xbridge 150*25mm*5um; mobile phase: [water (NH ₄ HCO ₃ )-acetonitrile]; gradient: 36%-66% B over 9 min, 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-d3)-5-oxyylidenepyrrolidine-2-methamide (I-20, 0.75 mg, yield 3.64%).

¹ H NMR (400 MHz, 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 synthesis route is as follows:

Step 1: Methyl 4-(trifluoromethyl)picolinate

2-Bromo-4-(trifluoromethyl)pyridine (30.0 g, 132 mmol) was dissolved in methanol (300 mL), and triethylamine (26.8 g, 265 mmol) and [1,1 -Bis(diphenylphosphorus)ferrocene]palladium dichloride (1.94 g, 2.65 mmol), the reaction solution was replaced three times with carbon monoxide gas, and then stirred for 12 hours under carbon monoxide (50.0 Psi) pressure and 80°C. After the reaction is completed, filter directly and concentrate under reduced pressure to obtain a brown liquid, then add water (500 mL) to dilute, extract with ethyl acetate (300 mL*3), and wash the organic phase with saturated sodium chloride solution (500 mL) and anhydrous sulfuric acid. After drying over sodium, it was filtered and concentrated to obtain methyl 4-(trifluoromethyl)picolinate (27.2 g, crude product).

LCMS, M/Z (ESI): 206.1 [M+H] ⁺ .

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

Dissolve methyl 4-(trifluoromethyl)methylpyridine ester (10.0 g, 48.7 mmol) in ethanol (100 mL), and then add sodium borohydride (3.69 g, 97.5 mmol), the reaction solution was stirred at 25°C for 12 hours. After the reaction is completed, slowly add saturated aqueous ammonium chloride solution (50 mL) at 0°C for quenching, then extract with ethyl acetate (100 mL*3), and wash the organic phase with saturated sodium chloride solution (200 mL) After drying with anhydrous sodium sulfate, it was filtered and concentrated to obtain (4-(trifluoromethyl)pyridin-2-yl)methanol (4.00 g, crude product), which was directly used in the next step.

¹ H NMR (400 MHz, CHCl ₃ -d) δ 8.63 (d, 1 H), 7.54 (s, 1 H), 7.34 (d, 1H), 4.78 (s, 2 H).

Step 3: 4-(trifluoromethyl)picolinaldehyde

Dissolve (4-(trifluoromethyl)pyridin-2-yl)methanol (4.00 g, 22.6 mmol) in tetrahydrofuran (40.0 mL), then add manganese dioxide (19.6 g, 225 mmol) in batches, and react The solution was stirred at 70°C for 1 hour. After the reaction was completed, the reaction solution was directly filtered and concentrated under reduced pressure to obtain 4-(trifluoromethyl)picolinal (1.40 g, crude product), which was directly used in the next step.

¹ H NMR (400 MHz, CHCl ₃ -d) δ 10.14 (s, 1H), 9.00 (d, 1H), 8.18 (s, 1H), 7.64 - 7.88 (m, 1H), 7.76 (dd, 0.94 Hz, 1H).

Step 4: 2-(difluoromethyl)-4-(trifluoromethyl)pyridine2-(difluoromethyl)-4-(trifluoromethyl)pyridine

Dissolve 4-(trifluoromethyl)picolinaldehyde (1.50 g, 18.5 mmol) in dichloromethane (10.0 mL), then add bis(2-methoxyethyl)amine at 0°C Sulfur trifluoride (3.75 g, 37.1 mmol). The reaction solution was stirred at 25°C for 2 hours. After the reaction is completed, add saturated sodium bicarbonate solution at 0°C to quench the reaction until bubbles no longer occur, then extract with dichloromethane (10 mL), combine the organic phases to obtain 2-(difluoromethyl)- 4-(Trifluoromethyl)pyridine (1.00 g, crude) was used directly in the next step.

Step 5: 2-(difluoromethyl)-4-(trifluoromethyl)pyridine 1-oxide2-(difluoromethyl)-4-(trifluoromethyl)pyridine 1-oxide

Dissolve 2-(difluoromethyl)-4-(trifluoromethyl)pyridine (2.00 g, 10.1 mmol) in dichloromethane (20.0 mL), and then add trifluoroacetic acid (5.78 g, 50.7 mmol) and hydrogen peroxide (71.0 mmol, 6.82 mL, 30% purity), and the reaction solution was stirred at 25°C for 12 hours. After the reaction is completed, add saturated sodium sulfite (10 mL) to quench the reaction, then extract with dichloromethane (10 mL*3), wash the organic phase with saturated sodium chloride solution (20 mL), dry with anhydrous sodium sulfate, filter and concentrate. Get crude product. The crude product was separated and purified by column chromatography (mobile phase was petroleum ether: ethyl acetate = 5:1 - 3:1) to obtain 2-(difluoromethyl)-4-(trifluoromethyl)pyridine 1-oxide ( 600 mg, yield 27.8%).

¹ H NMR (400 MHz, CHCl ₃ - d ) δ 8.33 (d, 1H), 7.86 (d, 1H), 7.62 (dd, 2.06 Hz, 1H), 7.22 (t, 1H).

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

Dissolve 2-(difluoromethyl)-4-(trifluoromethyl)pyridine 1-oxide (160 mg, 505 umol) in phosphorus oxychloride (3.60 g, 23.4 mmol). The reaction solution is heated at 100°C. Stir for 3 hours. After the reaction is completed, the reaction solution is slowly added to water (10.0 mL) to quench, then washed with saturated sodium bicarbonate, extracted with ethyl acetate (10.0 mL*2), and the organic phase is quenched with saturated sodium chloride solution (10 mL ), washed and dried over anhydrous sodium sulfate, filtered and concentrated at 30°C to obtain 2-chloro-6-(difluoromethyl)-4-(trifluoromethyl)pyridine (B21-1) (150 mg, crude product ), used directly in the next step.

Step 7: tertiary butyl(3aR,6S,6aS)-6-((5-chloro-2,4-difluorophenyl)(methyl-d3)aminocarbonyl)-2,2,3a- Trimethyltetrahydro-5H-[1,3]dioxola[4,5-c]pyrrole-5-carboxylate (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(3aR,6S,6aS)-6-((5-chloro-2,4-difluorophenyl)aminocarbonyl)-2,2,3a-trimethyltetrahydro-5H-[ 1,3]dioxalan[4,5-c]pyrrole-5-carboxylate (B5-7) (3.20 g, 7.16 mmol) and cesium carbonate (4.67 g, 14.32 mmol) were dissolved in N,N -In dimethylformamide (30.0 mL), the reaction solution was stirred at room temperature for 30 minutes, then deuterated methyl iodide (3.56 g, 25.06 mmol) was slowly added dropwise, and the reaction solution was stirred at 25°C for 2 hours. After the reaction was completed, water (200 mL) was added to dilute the reaction, and then extracted with ethyl acetate (30 mL*3). The organic phase was washed with saturated sodium chloride solution (60 mL) and dried over anhydrous sodium sulfate, then filtered and concentrated to obtain the third Tributyl(3aR,6S,6aS)-6-((5-chloro-2,4-difluorophenyl)(methyl-d3)aminocarbonyl)-2,2,3a-trimethyltetrahydro -5H-[1,3]dioxalan[4,5-c]pyrrole-5-carboxylate (B21-2) (2.00 g, crude product) was used directly in the next step.

LC-MS, M/Z (ESI): 364.1[M+H] ⁺ .

Step 8: tertiary butyl(3aS,6S,6aS)-6-((5-chloro-2,4-difluorophenyl)(methyl-d3)aminocarbonyl)-2,2,3a- Trimethyl-4-oxyylidenetetrahydro-5H-[1,3]dioxola[4,5-c]pyrrole-5-carboxylate (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

Under nitrogen protection, dissolve ruthenium trichloride hydrate (291 mg, 1.29 mmol) and sodium periodate (2.77 g, 12.9 mmol) in water (30.0 mL), replace with nitrogen three times, and stir the reaction solution at 25°C. 30 minutes, then tert-butyl(3aR,6S,6aS)-6-((5-chloro-2,4-difluorophenyl)(methyl-d3) dissolved in ethyl acetate (30.0 mL) )Aminocarbonyl)-2,2,3a-trimethyltetrahydro-5H-[1,3]dioxalan[4,5-c]pyrrole-5-carboxylate (B21-2) ( 2.00 g, 4.31 mmol) was slowly added to the reaction solution, and finally, the reaction solution was stirred at 25°C for 12 hours. After the reaction is completed, add saturated sodium sulfite (30.0 mL) to quench the reaction, and then extract with ethyl acetate (30 mL*3). The organic phase is washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrate to obtain crude product. The crude product was separated and purified by column chromatography (mobile phase was petroleum ether: ethyl acetate = 5:1 to 1:1) to obtain tert-butyl (3aS, 6S, 6aS)-6-((5-chloro-2,4 -Difluorophenyl)(methyl-d3)aminocarbonyl)-2,2,3a-trimethyl-4-oxyylidenetetrahydro-5H-[1,3]dioxola[4, 5-c]pyrrole-5-carboxylate (B21-3) (1.00 g, yield 48.5%).

LC-MS, M/Z (ESI): 378.1[M+H] ⁺ .

Step 9: (3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-2,2,6a-trimethyl-N-(methyl-d3)-6- Oxylidene tetrahydro-4H-[1,3]dioxola[4,5-c]pyrrole-4-methamide (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(3aS,6S,6aS)-6-((5-chloro-2,4-difluorophenyl)(methyl-d3)aminocarbonyl)-2,2,3a-trimethyl -4-Oxylidenetetrahydro-5H-[1,3]dioxola[4,5-c]pyrrole-5-carboxylate (B21-3) (1.00 g, 2.09 mmol) was dissolved in di into methyl chloride (10.0 mL), then zinc bromide (2.36 g, 10.4 mmol) was added, and the reaction solution was stirred at 25°C for 12 hours. After the reaction is completed, quench the reaction with saturated sodium bicarbonate (100 mL), then extract with dichloromethane (50 mL*3), wash the organic phase with saturated sodium chloride solution (50 mL), dry with anhydrous sodium sulfate, and filter Concentrate to obtain (3aS, 4S, 6aS)-N-(5-chloro-2,4-difluorophenyl)-2,2,6a-trimethyl-N-(methyl-d3)-6-oxoacetate Tetrahydro-4H-[1,3]dioxalan[4,5-c]pyrrole-4-methamide (B21-4) (500 mg, crude product, yield 63.2%).

¹ H NMR (400 MHz, CHCl ₃ - d ) δ 7.61 – 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).

Step 10: (3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-5-(6-(difluoromethyl)-4-(trifluoromethyl)pyridine -2-yl)-2,2,6a-trimethyl-N-(methyl-d3)-6-oxyylidenetetrahydro-4H-[1,3]dioxola[4,5- c]pyrrole-4-methamide (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

(3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-2,2,6a-trimethyl-N-(methyl-d3)-6-oxyylidene Tetrahydro-4H-[1,3]dioxola[4,5-c]pyrrole-4-methamide (B21-4) (180 mg, 544 umol) and 2-chloro-6-methyl -4-(Trifluoromethyl)pyridine (B21-1) (122.58 mg, 529.40 μmol) was dissolved in 1,4-dioxane (2.00 mL), and then cesium carbonate (172.49 mg) was added under nitrogen protection. , 529.40 μmol), tris(dibenzylideneacetone)dipalladium (36.36 mg, 39.71 μmol) and 4,5-bis(diphenylphosphonium)-9,9-dimethylxanthene (45.95 mg, 79.41 μmol), after nitrogen replacement three times, the reaction solution was stirred at 100°C for 3 hours. After the reaction is completed, the crude product is directly filtered and concentrated to obtain a brown oily substance. The crude product is separated and purified by prep-TLC (the mobile phase is petroleum ether: ethyl acetate = 3:1). After purification, (3aS, 4S, 6aS) is obtained. -N-(5-chloro-2,4-difluorophenyl)-5-(6-(difluoromethyl)-4-(trifluoromethyl)pyridin-2-yl)-2,2,6a -Trimethyl-N-(methyl-d3)-6-oxyylidenetetrahydro-4H-[1,3]dioxalan[4,5-c]pyrrole-4-methamide (B21 -5) (60.0 mg, yield 37.5%).

LC-MS, M/Z (ESI): 573.1[M+H] ⁺ .

Step 11: (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-oxyylidenepyrrolidine-2-methamide (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

(3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-5-(6-(difluoromethyl)-4-(trifluoromethyl)pyridine-2- methyl)-2,2,6a-trimethyl-N-(methyl-d3)-6-oxyylidenetetrahydro-4H-[1,3]dioxola[4,5-c]pyrrole -4-Formamide (B21-5) (60.0 mg, 104 μmol) was dissolved in dichloromethane (1.00 mL), and a solution of boron trichloride in dichloromethane (1 M, 314 μL), the reaction solution was stirred at 25°C for 2 hours. After the reaction is completed, adjust the pH to 9 with saturated sodium bicarbonate solution, then extract with dichloromethane (10 mL*3), wash the organic phase with saturated sodium chloride solution (20.0 mL), dry with anhydrous sodium sulfate, filter and concentrate. Get crude product. The crude product was separated and purified by reversed-phase high performance liquid chromatography. The separation method was: Waters Xbridge 150*25mm*5um; mobile phase: [water (NH ₄ HCO ₃ )-acetonitrile]; B%: 28%-58%, 8min , after purification, (2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-1-(6-(difluoromethyl)-4-(trifluoromethyl)pyridine is obtained -2-yl)-3,4-dihydroxy-4-methyl-N-(methyl-d3)-5-oxyylidenepyrrolidine-2-methamide (I-21) (8.65 mg, product rate 16.5%).

¹ H NMR (400 MHz, CH ₃ OH- d ₄ ) δ 8.60- 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 synthesis route is as follows:

Step 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-carbonyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-methyl Synthesis of amide (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

(3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-2,2,6a-trimethyl-N-(methyl-d3)-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-bisdiphenylphosphine-9,9-dimethylxanthene (900 mg, 1.48 mmol), 2-chloro-6 -Methyl-4-(trifluoromethyl)pyridine (3.5 g, 17.76 mmol) was dissolved in 1,4-dioxane (80 mL) solution. The reaction system was replaced with argon three times, and then reacted at 100 °C for 12 h. The reaction solution was filtered, and the filtrate was washed with water, dried, and concentrated to dryness to obtain a crude product, which was purified by a silica gel column (ethyl acetate/petroleum ether = 1/5) to obtain (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-Carbonyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-methamide (B22-1) (5.1 g, yield 63.7%).

LC-MS, M/Z (ESI): 537.1 [M+H] ⁺ .

Step 2: (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-carbonylpyrrolidine-2-methamide (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 nitrogen protection, (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-carbonyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4- Formamide (B22-1) (5.1 g, 9.5 mmol) was dissolved in dichloromethane (50 mL), and then boron trichloride (BCl ₃ ) in dichloromethane (19 mL, 19 mmol, 1 mol/L). The reaction solution was continued to stir at 0°C for 2 h. After the reaction was completed, the reaction was quenched with saturated sodium bicarbonate solution and adjusted to pH=8-9. Use (dichloromethane/methanol=20/1, 30 mL *2) After extraction with the mixed solvent, the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by HPLC to obtain (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-carbonylpyrrolidine-2-methyl Amide (I-22) (2.7 g, yield 57.4%).

¹ H NMR (400 MHz, CH ₃ OH-d4) δ 8.57–8.39 (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 synthesis route is as follows:

Step one: 6-chloro-3-fluoro-4-iodo-2-methylpyridine

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

Dissolve 6-chloro-3-fluoro-2-methyl-pyridine (3.00 g, 20.6 mmol) in tetrahydrofuran (30.0 mL), and add lithium diisopropylamide (2.5 M under nitrogen atmosphere at -78 °C , 10.7 mL), the reaction solution was stirred at -78°C for 30 minutes, then iodine element (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°C conditions for 1.5 hours. After the reaction is completed, slowly drop the reaction solution into the saturated ammonium chloride solution at 0°C, then extract with ethyl acetate (20 mL*3), and wash the organic phase with saturated sodium chloride solution (50 mL). After drying over anhydrous sodium sulfate, filter and concentrate to obtain a yellow solid crude product. The crude product is separated and purified by column chromatography (mobile phase is petroleum ether: ethyl acetate = 100:1 - 5:1) to obtain 6-chloro-3-fluoro-4. -Iodo-2-methylpyridine (3.00 g, yield 53.6%).

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.85 (d, 1 H) 2.41 (d, 3 H)

Step 2: 6-chloro-3-fluoro-2-methyl-4-(trifluoromethyl)pyridine6-chloro-3-fluoro-2-methyl-4-(trifluoromethyl)pyridine

Dissolve 6-chloro-3-fluoro-4-iodo-2-methylpyridine (4.00 g, 14.7 mmol) in N,N-dimethylformamide (80.0 mL), then add iodine at 25°C Cuprous chloride (8.42 g, 44.2 mmol) and methyl 2,2-difluoro-2-fluorosulfonyl-acetate (11.3 g, 58.9 mmol) were stirred at 100°C for 3 hours. After the reaction is completed, add water (100 mL) to dilute, then extract with ethyl acetate (200 mL*3), wash the organic phase with saturated sodium chloride solution (60.0 mL) and dry over anhydrous sodium sulfate, filter and concentrate to obtain 6 -Chloro-3-fluoro-2-methyl-4-(trifluoromethyl)pyridine (2.50 g, crude product), the crude product was used directly in the next step. LC-MS, M/Z (ESI): 214.1[M+H] ⁺ .

Step 3: (3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-5-[5-fluoro-6-methyl-4-(trifluoromethyl)pyridine -2-yl]-N-(²H₃)methyl-2,2,6a-trimethyl-6-oxyylidene-hexahydro-[1,3]dioxola[4,5-c] Pyrrole-4-methamide (B23-2) (3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-5-[5-fluoro-6-methyl-4-(trifluoromethyl) pyridin-2-yl]-N-(²H₃)methyl-2,2,6a-trimethyl-6-oxo-hexahydro-[1,3]dioxolo[4,5-c]pyrrole-4-carboxamide

(3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-2,2,6a-trimethyl-N-(methyl-d3)-6-oxyylidene Tetrahydro-4H-[1,3]dioxola[4,5-c]pyrrole-4-methamide (B21-4) (300 mg, 794 μmmol) and 6-chloro-3-fluoro- 2-Methyl-4-(trifluoromethyl)pyridine (B23-1) (508 mg, 2.38 mmol) was dissolved in 1,4-dioxane (3.00 mL), and then carbonic acid was added under nitrogen protection. Cesium (517 mg, 1.59 mmol), tris(dibenzylideneacetone)dipalladium (109 mg, 119 μmol), and 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene ( 109 mg, 119 μmol), after nitrogen replacement three times, the reaction solution was stirred at 90°C for 12 hours. After the reaction is completed, the crude product is directly filtered and concentrated to obtain a brown oily substance. The crude product is separated and purified by prep-TLC (the mobile phase is petroleum ether: ethyl acetate = 3:1). After purification, the crude product is obtained, and the crude product is separated and purified by reverse-phase high-efficiency Liquid chromatography was used for separation and purification. The analysis method was: WatersXbridge 150*25mm*5um; mobile phase: [water(NH ₄ HCO ₃ )-ACN]; gradient: 60%-90% B over 9 min. Obtain (3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-5-(6-(difluoromethyl)-4-(trifluoromethyl)pyridine-2- methyl)-2,2,6a-trimethyl-N-(methyl-d3)-6-oxyylidenetetrahydro-4H-[1,3]dioxalan[4,5-c]pyrrole -4-Formamide (B23-2) (80.0 mg, yield 18.6%). LC-MS, M/Z (ESI): 555.0[M+H] ⁺ .

Step 4: (2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-1-[5-fluoro-6-methyl-4-(trifluoromethyl)pyridine -2-yl]-3,4-dihydroxy-N-(²H₃)methyl-4-methyl-5-oxyylidenepyrrolidine-2-methamide (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-(²H₃) Methyl-4-methyl-5-oxopyrrolidine-2-carboxamide I-23

(3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-5-(6-(difluoromethyl)-4-(trifluoromethyl)pyridine-2- methyl)-2,2,6a-trimethyl-N-(methyl-d3)-6-oxyylidenetetrahydro-4H-[1,3]dioxalan[4,5-c]pyrrole -4-Formamide (B23-2) (60.0 mg, 108 μmol) was dissolved in dichloromethane (1.00 mL), and a solution of boron trichloride in dichloromethane (1 M, 324 μL), the reaction solution was stirred at 25°C for 2 hours. After the reaction is completed, adjust the pH to 9 with saturated sodium bicarbonate solution, then extract with dichloromethane (10 mL*3), wash the organic phase with saturated sodium chloride solution (20.0 mL), dry with anhydrous sodium sulfate, filter and concentrate. Get crude product. The crude product was separated and purified _by reversed-phase high performance liquid chromatography. The separation _method was: Waters , after purification, (2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-1-[5-fluoro-6-methyl-4-(trifluoromethyl)pyridine is obtained -2-yl]-3,4-dihydroxy-N-(²H₃)methyl-4-methyl-5-oxyylidenepyrrolidine-2-methamide (I-23) (43.0 mg, yield 77.2%).

¹ H NMR (400 MHz, CH ₃ OH- d ₄ ) δ 8.45 - 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 synthesis route is as follows:

Step 1: 2-Bromo-6-(²H₃)methyl-4-(trifluoromethyl)pyridine

2-bromo-6-(²H₃)methyl-4-(trifluoromethyl)pyridine B24-1

Dissolve 2,6-dibromo-4-(trifluoromethyl)pyridine (2.00 g, 6.56 mmol) in tetrahydrofuran (60.0 mL) and N-methylpyrrolidone (4.00 mL), and add acetyl under nitrogen atmosphere Iron acetonate (231.67 mg, 655.97 μmol) and deuterated methane magnesium iodide (1 M, 11.81 mL) were stirred at 25°C for 1 hour. After the reaction is completed, quench with saturated ammonium chloride (30.0 mL), extract with ethyl acetate (300 mL*3), wash with saturated sodium chloride solution (500 mL) and dry over anhydrous sodium sulfate, filter and concentrate to obtain 2 -Bromo-6-(²H₃)methyl-4-(trifluoromethyl)pyridine (B24-1) (27.2 g, crude).

LCMS, M/Z (ESI): 244.9 [M+H] ⁺ .

Step 2: (3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-N-(²H₃)methyl-2,2,6a-trimethyl-5-[ 6-(²H₃)methyl-4-(trifluoromethyl)pyridin-2-yl]-6-oxyylidene-hexahydro-[1,3]dioxola[4,5-c]pyrrole -4-Formamide (B24-2) (3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-N-(²H₃)methyl-2,2,6a-trimethyl-5- [6-(²H₃)methyl-4-(trifluoromethyl)pyridin-2-yl]-6-oxo-hexahydro-[1,3]dioxolo[4,5-c]pyrrole-4-carboxamide B24-2

(3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-2,2,6a-trimethyl-N-(methyl-d3)-6-oxyylidene Tetrahydro-4H-[1,3]dioxola[4,5-c]pyrrole-4-methamide (B21-4) (300 mg, 794 μmol) and 2-chloro-6-methyl -4-(Trifluoromethyl)pyridine (578 mg, 2.38 mmol) was dissolved in 1,4-dioxane (3.00 mL), and then cesium carbonate (300 mg, 794 μmol) and cesium carbonate (300 mg, 794 μmol) were added under nitrogen protection. Tris(dibenzylideneacetone)dipalladium (109 mg, 119 μmol) and 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (137 mg, 238 μmol), nitrogen replacement After three times, the reaction solution was stirred at 100°C for 12 hours. After the reaction is completed, the crude product is directly filtered and concentrated to obtain a brown oily substance. The crude product is separated and purified by column chromatography (mobile phase is petroleum ether: ethyl acetate = 3:1). After purification, (3aS, 4S, 6aS) is obtained. -N-(5-chloro-2,4-difluorophenyl)-N-(²H₃)methyl-2,2,6a-trimethyl-5-[6-(²H₃)methyl-4-( Trifluoromethyl)pyridin-2-yl]-6-oxyylidene-hexahydro-[1,3]dioxola[4,5-c]pyrrole-4-methamide (B24-2) (60.0 mg, yield 14.0%).

LC-MS, M/Z (ESI): 540.3[M+H] ⁺ .

Step 3: (2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-3,4-dihydroxy-N-(²H₃)methyl-4-methyl-1 -[6-(²H₃)methyl-4-(trifluoromethyl)pyridin-2-yl]-5-oxyylidenepyrrolidine-2-methamide (I-24) (2S,3S,4S) -N-(5-chloro-2,4-difluorophenyl)-3,4-dihydroxy-N-(²H₃)methyl-4-methyl-1-[6-(²H₃)methyl-4-(trifluoromethyl)pyridin-2 -yl]-5-oxopyrrolidine-2-carboxamide I-24

(3aS,4S,6aS)-N-(5-chloro-2,4-difluorophenyl)-N-(²H₃)methyl-2,2,6a-trimethyl-5-[6-( ²H₃)methyl-4-(trifluoromethyl)pyridin-2-yl]-6-oxyylidene-hexahydro-[1,3]dioxalan[4,5-c]pyrrole-4- Formamide (B24-2) (60.0 mg, 111 μmol) was dissolved in dichloromethane (1.00 mL), and then a solution of boron trichloride in dichloromethane (1 M, 333 μL) was added at -20°C. The reaction solution was stirred at 25°C for 2 hours. After the reaction is completed, adjust the pH to 9 with saturated sodium bicarbonate solution, then extract with dichloromethane (10 mL*3), wash the organic phase with saturated sodium chloride solution (20.0 mL), dry with anhydrous sodium sulfate, filter and concentrate. Get crude product. The crude product was separated and purified by reversed-phase high performance liquid chromatography. The separation method was: Waters Xbridge 150*25mm*5um; mobile phase: [water (NH ₄ HCO ₃ )-acetonitrile]; B%: 28%-58%, 8min , after purification, (2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-3,4-dihydroxy-N-(²H₃)methyl-4-methyl-1 -[6-(²H₃)methyl-4-(trifluoromethyl)pyridin-2-yl]-5-oxyylidenepyrrolidine-2-methamide (I-24) (54.0 mg, yield 97.2 %).

¹ H NMR (400 MHz, CH ₃ OH- d ₄ ) δ ppm 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 preparation of the following compounds can be obtained by referring to the preparation methods of Examples 1-10: Structural formula name LC-MS, M/Z (ESI) (2S)-N-(5-chloro-2,4-difluorophenyl)-N-methyl-3-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-4 -Oxo-3-azabicyclo[3.1.0]hexane-2-methamide 460.0 (S)-N-(5-chloro-2,4-difluorophenyl)-N,2-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl )-5-oxopyrrolidine-2-methamide 462.1 (S)-N-(5-chloro-2,4-difluorophenyl)-N,4,4-trimethyl-1-(6-methyl-4-(trifluoromethyl)pyridine-2 -yl)-5-oxopyrrolidine-2-methamide 476.1 (6S,7R)-N-(5-chloro-2,4-difluorophenyl)-7-hydroxy-N-methyl-5-(6-methyl-4-(trifluoromethyl)pyridine- 2-yl)-4-oxo-5-azaspiro[2.4]heptane-6-methamide 490.1 (2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-3,4-dihydroxy-N,3-dimethyl-1-(6-methyl-4- (Trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-methamide 494.1 (2S,3R)-N-(5-chloro-2,4-difluorophenyl)-3-cyano-N-methyl-1-(6-methyl-4-(trifluoromethyl)pyridine -2-yl)-5-oxopyrrolidine-2-methamide 473.1 (2S,3S,4S)-1-(4,6-bis(trifluoromethyl)pyridin-2-yl)-N-(5-chloro-2,4-difluorophenyl)-3,4- Dihydroxy-N,4-dimethyl-5-oxopyrrolidine-2-methamide 548.0 (2S,4S)-N-(5-chloro-2,4-difluorophenyl)-3,4-dihydroxy-N-methyl-1-(6-methyl-4-(trifluoromethyl) )pyridin-2-yl)-5-oxo-4-(trifluoromethyl)pyrrolidine-2-carboxamide 548.05 (2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-3,4-dihydroxy-N,3,4-trimethyl-1-(6-methyl- 4-(Trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-methamide 508.10 (2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-4-cyano-3,4-dihydroxy-N-methyl-1-(6-methyl- 4-(Trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-methamide 505.06 (2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-4-(difluoromethyl)-3,4-dihydroxy-N-methyl-1-(6 -Methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-methamide 530.06 (2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-1-(6-(difluoromethyl)-4-(trifluoromethyl)pyridin-2-yl )-3,4-dihydroxy-N,4-dimethyl-5-oxopyrrolidine-2-methamide 530.0 (2S,3S,4S)-N-(3-fluoro-1H-pyrrole[2,3-b]pyridin-6-yl)-3,4-dihydroxy-N,4-dimethyl-1-( 6-Methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-methamide 482.1 (2S,3S,4S)-1-(4,6-bis(trifluoromethyl)pyrimidin-2-yl)-N-(5-chloro-2,4-difluorophenyl)-3,4- Dihydroxy-N,4-dimethyl-5-oxopyrrolidine-2-methamide 549.1 ( R )- N -(5-chloro-2,4-difluorophenyl)-N-deuteromethyl-5-(6-methyl-4-(trifluoromethyl)pyridin-2-yl) -4-oxo-5-azaspiro[2.4]heptane-6-formamide 476.8 (S)-N-(5-chloro-2,4-difluorophenyl)-N-deuteromethyl-5-(6-methyl-4-trifluoromethyl)pyridin-2-yl)- 4-oxo-5-azaspiro[2.4]heptane-6-formamide 476.8 (2S)-N-(5-chloro-2,4-difluorophenyl)-N-deuteromethyl-3-(6-methyl-4-(trifluoromethyl)pyridin-2-yl) -4-Oxo-3-azabicyclo[3.1.0]hexane-2-methamide 463.0 (S)-N-(5-chloro-2,4-difluorophenyl)-N-deuteromethyl-2-methyl-1-(6-methyl-4-(trifluoromethyl)pyridine -2-yl)-5-oxopyrrolidine-2-methamide 465.1 (S)-N-(5-chloro-2,4-difluorophenyl)-4,4-difluoro-N-deuteratedmethyl-1-(6-methyl-4-(trifluoromethyl) )pyridin-2-yl)-5-oxopyrrolidine-2-methamide 487.1 (S)-N-(5-chloro-2,4-difluorophenyl)-N-deuteromethyl-4,4-dimethyl-1-(6-methyl-4-(trifluoromethyl (yl)pyridin-2-yl)-5-oxopyrrolidine-2-methamide 479.1 (6S,7R)-N-(5-chloro-2,4-difluorophenyl)-7-hydroxy-N-deuteratedmethyl-5-(6-methyl-4-(trifluoromethyl) Pyridin-2-yl)-4-oxo-5-azaspiro[2.4]heptane-6-methamide 493.1 (2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl) –N-deuteratedmethyl-3,4-dihydroxy-3-methyl-1-(6-methyl yl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-carboxamide 497.1 (2S,3R)-N-(5-chloro-2,4-difluorophenyl)-3-cyano-N-deuteratedmethyl-1-(6-methyl-4-(trifluoromethyl) )pyridin-2-yl)-5-oxopyrrolidine-2-methamide 476.1 (2S,4S)-N-(5-chloro-2,4-difluorophenyl)-N-deuteratedmethyl-4-hydroxy-4-methyl-1-(6-methyl-4-( Trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-methamide 481.09 (2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-4-cyclopropyl-3,4-dihydroxy-N-deuteratedmethyl-1-(6- Methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-carboxamide 523.10 (2S,3S,4S)-1-(4,6-bis(trifluoromethyl)pyridin-2-yl)-N-(5-chloro-2,4-difluorophenyl)-N-deuterated Methyl-3,4-dihydroxy-4-methyl-5-oxopyrrolidine-2-methamide 551.0 (2S,4S)-N-(5-chloro-2,4-difluorophenyl)-3,4-dihydroxy-N-deuteratedmethyl-1-(6-methyl-4-(trifluoro Methyl)pyridin-2-yl)-5-oxo-4-(trifluoromethyl)pyrrolidine-2-methamide 551.05 (2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-N-deuteratedmethyl-3,4-dihydroxy-3,4-trimethyl-1-( 6-Methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-methamide 511.10 (2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-4-cyano-3,4-dihydroxy-N-deuteratedmethyl-1-(6-methyl yl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-methamide 508.06 (2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-4-(difluoromethyl)-3,4-dihydroxy-N-deuteratedmethyl-1- (6-Methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-methamide 533.06 (2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-1-(6-(difluoromethyl)-4-(trifluoromethyl)pyridin-2-yl )-N-deuteromethyl-3,4-dihydroxy-4-methyl-5-oxopyrrolidine-2-methamide 533.0 (2S,3S,4S)-N-(3-fluoro-1H-pyrrole[2,3-b]pyridin-6-yl)-N-deuteromethyl-3,4-dihydroxy-4-methyl -1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-methamide 485.1 (2S,3S,4S)-1-(4,6-bis(trifluoromethyl)pyrimidin-2-yl)-N-(5-chloro-2,4-difluorophenyl)-N-deuterated Methyl-3,4-dihydroxy-4-methyl-5-oxopyrrolidine-2-methamide 552.1 N-(5-chloro-2,4-difluorophenyl)-1-(4-cyclopropyl-6-methylpyridin-2-yl)-N-deuteratedmethyl-3,4-dihydroxy -4-Methyl-5-oxopyrrolidine-2-methamide 469.13 (2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-1-[5-fluoro-6-methyl-4-(trifluoromethyl)pyridin-2-yl ]-3,4-dihydroxy-N-methyl-4-methyl-5-oxyylidenepyrrolidine-2-methamide 511.07 (2S,3S,4S)-N-(5-chloro-2,4-difluorophenyl)-3,4-dihydroxy-N-methyl-4-methyl-1-[6-(²H₃) Methyl-4-(trifluoromethyl)pyridin-2-yl]-5-oxyylidenepyrrolidine-2-methamide 496.10

Effect test example 1

In the test examples of the present invention, the preparation method of comparative compound I refers to patent WO2021/028643A1, and the preparation method of comparative compound II refers to patent WO2021028670A1. The structures are as follows. Control Compound I Control Compound II

Test Example 1: Compound inhibition test on Polθ polymerase activity

The recombinant Polθ polymerase domain (aa 1819-2590) used in this test was expressed and purified using Invitrogen's Bac-to-Bac Baculovirus Expression System. The detection method used to measure Polθ polymerase activity is the PicoGreen dsDNA quantitative assay.

The reaction system consists of compounds dissolved in dimethylsulfoxide (DMSO), purified recombinant Polθ (aa 1819-2590) protein, adhesive mixture and dNTPs. All compounds were first prepared into 10mM stock solutions with dimethylsulfoxide (DMSO), and were treated with experimental buffer (25mM trishydroxymethylaminomethane hydrochloride (Tris HCl) pH 7.5, 12.5mM chloride) before use. Sodium (NaCl), 0.5 mM magnesium chloride (MgCl ₂ ), 5% glycerol, 0.01% Triton X-100, 0.01% bovine gamma globulin (BGG), and 1 mM dithiothreitol (DTT)) in a gradient dilution; reconstituted The protein solution is recombinant Polθ (aa 1819-2590) protein with a final concentration of 30nM dissolved in experimental buffer; the binding mixture is a primer with a final concentration of 150nM (5' - GCG GCT GTC ATA AG - 3') and a final concentration of 150nM. The template (5'-GCT ACA TTG ACA ATG GCA TCA AAT CTC AGA TTG CGT CTT ATG ACA GCC) was dissolved in the experimental buffer, heated to 42°C for 3 minutes and then bonded; the dNTP solution was dNTP with a final concentration of 120 μM dissolved in the experimental buffer. Obtained from buffer solution.

In the compound screening experiment, first add 2 μL/well of the recombinant protein solution to the 384-well plate (Perkin Elmer-Proxiplate), then add 0.06 μL/well of different concentrations of compounds and negative control (DMSO) in sequence, and finally add 2 μL/well of the recombinant protein solution. Adhesion mix and 2 μL/well of dNTPs. The blank control group contained only the adhesive mixture and dNTPs. After sealing the 384-well plate with aluminum foil sealing film, place it at 37°C and incubate for 30 minutes. Then add 4 μL of reaction stop solution (25mM tris-hydroxymethylaminomethane hydrochloride (Tris-HCl), 10mM) to the test well. The reaction was terminated with ethylenediaminetetraacetic acid (EDTA) and 1:80 PicoGreen), and after incubation at room temperature in the dark for 90 minutes, the reaction was read on an enzyme label reader (PerkinElmer EnVision®2104: λex =485nm λem =520nm) Fluorescence value. The inhibition rate was calculated as follows: Inhibition rate = (1-(compound group-blank group)/(negative control group-blank group)) × 100%. Use GraphPad Prism8 software to calculate the IC ₅₀ value of each compound. The results are shown in Table 1.

Table 1. Results of compounds inhibiting Polθ polymerase activity test compound 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 twenty two I-23 twenty one I-24 45

Experimental results show that the compound of the present invention has good inhibitory effect on Polθ polymerase.

test case 2. Test of compounds inhibiting tumor cell proliferation

DLD-1BRCA2(-/-) cells were seeded in a 96-well cell plate at a density of 500 cells/100 μL/well, and the cell plate was placed in an incubator for 18 hours (37°C, 5% CO ₂ ). Start drug treatment on the next day, add 100 μL/well gradient diluted test compound solution to the culture plate culture medium (the starting concentration of each drug is 100 μM, the diluent is dimethylsulfoxide (DMSO), and the dilution ratio is 1 :3, each drug is diluted to nine gradient points) or dimethylsulfoxide (DMSO) (negative control), and a blank group is set up without inoculating cell lines and only adding culture medium. After adding the drug, continue to place the culture plate in culture Incubate for 5 days in a box (37°C, 5% CO ₂ ). On the fourth and seventh days respectively, fresh medium and compounds were replaced. For detection on the tenth day, add 100 μL CellTiter-Glo (Promega G9243) reagent to each well, shake for 5 minutes and then let stand at room temperature for 5 minutes. Use an enzyme label reader (PerkinElmer EnVision® 2104) to measure the chemistry of each well. Luminous signal value. Cell proliferation inhibition rate = (1-(compound group-blank group)/(DMSO group-blank group)) × 100%. Use GraphPad Prism8 software to calculate the IC ₅₀ value of each compound. The results are shown in Table 2.

Table 2. Inhibitory effects of compounds on tumor cell proliferation test compound IC ₅₀ (μM) Control Compound II 15 I-5 3.31 I-21 5.54 I-22 4.96

Experimental results show that the compounds of the present invention exhibit inhibitory effects on the proliferation of DLD-1 BRCA2 (-/-) cells.

test case 3 : Mouse pharmacokinetic test

Use male ICR mice, 20-25g, fasted overnight. Take 3 mice and administer a dose of 10 mg/kg orally and a dosage volume of 10 mL/kg. In addition, 3 mice were taken, and the intravenous dosage was 3 mg/kg, and the dosage volume was 5 mL/kg. Blood was collected before administration and at 5, 15, 30 minutes, and 1, 2, 4, 6, 8, and 24 hours after administration. The blood sample is 6800g, centrifuged at 6000g for 3 minutes at 2-8°C, the plasma is separated and collected, and stored at -80°C. Take the plasma at each time point, add 3-5 times the amount of acetonitrile solution containing the internal standard, mix, vortex and mix for 1 minute, 13000 rpm, centrifuge at 4°C for 10 minutes, take the supernatant, add 3 times the amount of water, mix, take an appropriate amount and mix The liquid was analyzed by LC-MS/MS. Using blood drug concentration data at different time points, Phoenix WinNonlin8.2.0 is used to calculate pharmacokinetic parameters, providing CL, Vz, AUC _0-t , AUC _{0- ∞} , MRT _{0- ∞} , Cmax, Tmax, and T _{1/ 2} and other parameters and their mean and standard deviation. The results are shown in Tables 3 and 4.

Table 3. Mouse pharmacokinetic test results of test compounds compound Mouse pharmacokinetic parameters (intravenous administration) CL (L/h/kg) Vz (L/kg) AUC _0-t (h*ng/mL) T _1/2 (hr) Control Compound II 0.33 0.85 8892 1.80 I-5 0.15 0.29 21576 1.53 I-21 0.08 0.28 41509 2.55 I-22 0.13 0.40 23790 2.14

Table 4. Mouse pharmacokinetic test results of test compounds compound Mouse pharmacokinetic parameters (oral administration) Cmax (ng/mL) Tmax(hr) AUC _0-t (h*ng/mL) T _1/2 (hr) Control Compound II 7623 1 33533 2.67 I-5 15967 0.58 60343 2.69 I-21 24800 1 308975 2.81 I-22 14433 0.50 56115 2.05

Experimental results show that the compound of the present invention has high oral exposure, good pharmacokinetic properties and good druggability.

Although specific embodiments of the present invention have been described above, those skilled in the art should understand that these are only examples, and various changes or modifications can be made to these embodiments without departing from the principles and essence of the present invention. . Therefore, the protection scope of the present invention is limited by the appended patent application scope.

without.

Claims (20)

A heterocyclic compound represented by formula I, its stereoisomer or its pharmaceutically acceptable salt, 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 ₂ are each independently halogen, hydroxyl, Amino group, cyano group, unsubstituted or C ₁ -C ₆ alkyl group substituted by one or more R _a , unsubstituted or C ₁ -C ₆ alkoxy group substituted by one or more R _a , unsubstituted or 3-6 membered cycloalkyl substituted by one or more R _a ; in R ₁ and R ₂ , each R _a is independently deuterium, halogen, hydroxyl, amine, cyano, C ₁ -C ₆ alkyl Or C ₁ -C ₆ haloalkyl; R ₃ is hydrogen, unsubstituted or C ₁ -C ₆ alkyl substituted by one or more R _a ; in R ₃ , each R _a is independently deuterium, halogen, hydroxyl _{or _ _ _ _ _ _ _ _ _ _} , R ₄ , R ₅ , R ₆ , R ₇ and R ₈ are each independently hydrogen or R _b , and at least one of R ₄ , R ₅ , R ₆ , R ₇ and R ₈ is a cyano group; or, R ₄ and R ₅ are each independently R _b , or R ₆ and R ₇ are each independently R _b , or R ₄ and R ₅ together with the C atom to which they are connected form ring A, R ₆ , R ₇ and R ₈ is each independently hydrogen or R _b ; or, R ₅ and R ₆ together with the C atom to which they are connected form ring A, and R ₄ , R ₇ and R ₈ are each independently hydrogen or R _b ; or, R ₆ and R ₇ together with the C atom to which they are connected form Ring A, R ₄ , R ₅ and R ₈ are each independently hydrogen or R _b ; or, R ₇ and R ₈ together with the C atom to which they are connected form Ring A , R ₄ , R ₅ and R ₆ are each independently hydrogen or R _b ; each R _b is independently halogen, hydroxyl, amino, cyano, unsubstituted or C ₁ -C ₆ alkyl substituted by R _a , unsubstituted or substituted by R _a C ₁ -C ₆ alkoxy group, unsubstituted or substituted by R _a 3-6 membered cycloalkyl group, unsubstituted or substituted by R _a 4-6 membered heterocycloalkyl group ; In each R _b , each R _a is independently deuterium, halogen, hydroxyl, amine, cyano, C ₁ -C ₆ alkyl or C ₁ -C ₆ haloalkyl; each ring A is not 3-8 membered cycloalkyl substituted or substituted by one or more R _a , 4-8 membered heterocycloalkyl unsubstituted or substituted by R _a ; In each ring A, the R _a is independently deuterium , halogen, hydroxyl, amino, cyano, C ₁ -C ₆ alkyl or C ₁ -C ₆ haloalkyl. The heterocyclic compound represented by formula I, its stereoisomer or a pharmaceutically acceptable salt thereof as described in claim 1, wherein the heterocyclic compound represented by formula I satisfies the following conditions (1) to condition ( One or more conditions in 36): (1) In ring K, in the 5-6 membered heteroaromatic ring, the heteroatom is selected from one or more of 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, in the 5-6 membered heteroaromatic ring, the heteroatom is selected from N. More preferably, the The number of heteroatoms in the 5-6 membered heteroaromatic ring is 1; (2) In R ₁ and R ₂ , the halogen is each independently fluorine, chlorine, bromine or iodine, such as chlorine or fluorine; (3) R In ₁ and R ₂ , the C ₁ -C ₆ alkyl groups in the unsubstituted or substituted C ₁ -C ₆ alkyl groups with one or more R _a are each independently methyl, ethyl, n-propyl, iso Propyl, n-butyl, isobutyl or tert-butyl, such as methyl; (4) Among R ₁ and R ₂ , the C ₁ -C ₆ alkane that is unsubstituted or substituted by one or more R _a The C ₁ -C ₆ alkoxy groups in the oxygen group are each independently methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy or tert-butoxy; ( 5) In R ₁ and R ₂ , the 3-6 membered cycloalkyl groups are each independently cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, such as cyclopropyl; (6) R In ₁ and R ₂ , in each R _a , the halogen is independently fluorine, chlorine, bromine or iodine, such as fluorine; (7) In R ₁ and R ₂ , in each R _a , the C ₁ -C ₆ The alkyl groups are each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl; (8) In R ₁ and R ₂ , in each R _a , the The C ₁ -C ₆ haloalkyl groups are each independently a C 1 -C 6 alkyl group substituted by 1, 2 or 3 halogens; (9) In R ₃ , the said C ₁ -C ₆ haloalkyl group is unsubstituted or substituted by one or more R _a Each of the C ₁ -C ₆ alkyl groups in the substituted C ₁ -C ₆ alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, such as methyl (10) In R ₃ , in each R _a , the halogen is independently fluorine, chlorine, bromine or iodine, such as fluorine; (11) In R ₃ , in each R _a , the C ₁ -C ₆ The alkyl groups are each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl; (12) In R ₃ , in each R _a , the C ₁ -C ₆ haloalkyl is each independently a C ₁ -C ₆ alkyl substituted by 1, 2 or 3 halogens; (13) Among R ₄ , R ₅ , R ₆ , R ₇ and R ₈ , the C ₁ -C ₆ alkyl groups are each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, such as methyl; (14) In the R _b , The halogens are each independently fluorine, chlorine, bromine or iodine, such as _fluorine ; (15) In each R _b , the C ₁ -C ₆ alkyl in the unsubstituted or substituted C ₁ -C ₆ alkyl group The groups are each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, such as methyl; (16) In each R _b , the said unsubstituted or The C ₁ -C ₆ alkoxy groups in the C ₁ -C ₆ alkoxy group substituted by R _a are each independently methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutyl Oxy group or tert-butoxy group; (17) In each R _b , the 3-6 membered cycloalkyl group in the unsubstituted or substituted 3-6 membered cycloalkyl group by R _a is each independently a cyclopropyl group, Cyclobutyl, cyclopentyl or cyclohexyl, such as cyclopropyl; (18) In each R _b , 4-6 of the 4-6 membered heterocycloalkyl that is unsubstituted or substituted by R _a In the one-membered heterocycloalkyl group, the heteroatom is selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3; (19) In each R _b and each R _a , the halogens are each independent ground is fluorine, chlorine, bromine or iodine, such as fluorine; (20) In each R _b , in each R _a , the C ₁ -C ₆ alkyl group is each independently methyl, ethyl, n-propyl, iso Propyl, n-butyl, isobutyl or tertiary butyl; (21) In each R _b , in each R _a , the C ₁ -C ₆ haloalkyl groups are each independently 1, 2 or 3 Halogen-substituted C ₁ -C ₆ alkyl; (22) In each ring A, the 3-8-membered cycloalkyl group in the 3-8-membered cycloalkyl group that is unsubstituted or substituted by one or more R _a is 3 -6-membered cycloalkyl, for example, the 3-8-membered cycloalkyl is each independently cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, preferably cyclopropyl; (23) each In ring A, the 4-8-membered heterocycloalkyl group in the unsubstituted or R _a- substituted 4-8-membered heterocycloalkyl group is a 4-6-membered heterocycloalkyl group. Preferably, the 4-6-membered heterocycloalkyl group In the one-membered heterocycloalkyl group, the heteroatom is selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3; (24) In each ring A, in each R _a , the halogens are each independent ground is fluorine, chlorine, bromine or iodine; (25) In each ring A, in each R _a , the C ₁ -C ₆ alkyl groups are each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl; (26) In each ring A, in each R _a , the C ₁ -C ₆ haloalkyl groups are each independently substituted by 1, 2 or 3 halogens C ₁ -C ₆ alkyl; (27) When m is 2, 3, 4, the R ₁ is the same or different; (28) When n is 2, 3, 4, the R ₂ is the same or different; (29) When there are multiple R _a , the R _a is the same or different; (30) The "or" is only "or", expressing an alternative relationship between elements; (31) The alkyl group does not indicate substitution When, the alkyl group is an unsubstituted alkyl group; (32) When the alkyl group is not specified to be isotope-substituted, the alkyl group is an alkyl group that does not contain carbon and/or hydrogen isotopes; (33) The ring When the alkyl group or carbocyclic group is not specified as an unsaturated or partially saturated carbocyclic group, the cycloalkyl group or carbocyclic group is a saturated cycloalkyl group or carbocyclic group; (34) The heterocycloalkyl group is not specified. When it is an unsaturated or partially saturated heterocycloalkyl group, the heterocycloalkyl group is a saturated heterocycloalkyl group; (35) When the heteroaromatic ring group is not specified to be a heteroaromatic ring group fused with a benzene ring , the heteroaromatic ring group is an unfused heteroaromatic ring group; (36) the stereoisomer is a parapalpal isomer or a non-parapalpal isomer. The heterocyclic compound represented by formula I, its stereoisomer or a pharmaceutically acceptable salt thereof as described in claim 1, wherein the heterocyclic compound represented by formula I is any one of the following schemes: Scheme 1: In the heterocyclic compound represented by formula I, I, where m and n are respectively integers from 0 to 4; R ₁ and R ₂ are each independently selected from: halogen, hydroxyl, amino, cyano, C ₁ -C ₆ alkyl, 3-6 membered cycloalkyl group; the R ₁ or R ₂ is optionally substituted with a substituent selected from the following: halogen, hydroxyl, amino group, C ₁ -C ₆ alkyl group; when R ₁ is multiple, the R ₁ is the same or Different; when there are multiple R _2s , the R _2s are the same or different; R ₃ is hydrogen, C ₁ -C ₆ alkyl or C ₁ -C ₆ haloalkyl; R ₄ , R ₅ , R ₆ , R ₇ Each is independently hydrogen or C ₁ -C ₆ alkyl, R ₈ is C ₁ -C ₆ alkyl; or R ₄ and R ₅ together with the C atom to which they are connected form ring A, R ₆ , R ₇ , R ₈ is each independently hydrogen or C ₁ -C ₆ alkyl; or R ₅ and R ₆ together with the C atom to which they are connected form ring A, and R ₄ , R ₇ and R ₈ are each independently hydrogen or C ₁ - C ₆ alkyl; or R ₆ and R ₇ together with the C atoms to which they are connected form ring A, and R ₄ , R ₅ , and R ₈ are each independently hydrogen or C ₁ -C ₆ alkyl; or R ₇ and R ₈ together with the C atoms to which they are connected form Ring A, R ₄ , R ₅ and R ₆ are each independently hydrogen or C ₁ -C ₆ alkyl; the Ring A is a 3-8 membered cycloalkyl or 4- 8-membered heterocycloalkyl; The ring A is optionally substituted with a substituent selected from the following: halogen, hydroxyl, amine, cyano, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl; scheme 2: Among the heterocyclic compounds represented by formula I, I, where m is 0, 1, 2, 3, 4; n is 0, 1, 2, 3, 4; R ₁ and R ₂ are each independently selected from: halogen, hydroxyl, amino, cyano, un C ₁ -C ₆ alkyl substituted or substituted by one or more Ra, C ₁ -C ₆ alkoxy unsubstituted or substituted by one or more Ra, unsubstituted or substituted by one or more Ra 3 -6-membered cycloalkyl; when m is 2, 3, 4, the R ₁ is the same or different; when n is 2, 3, 4, the R ₂ is the same or different; R ₃ is hydrogen, unsubstituted Or C ₁ -C ₆ alkyl substituted by one or more _Ra ; R ₄ , R ₅ , R ₆ , R ₇ are each independently hydrogen or R _b , R ₈ is R _b ; or R ₄ , R ₅ , R ₆ , R ₇ , and R ₈ are each independently hydrogen or R _b , and at least one of R ₄ , R ₅ , R ₆ , R ₇ , and R ₈ is a cyano group; or R ₄ and R ₅ are the same or different R _b , or R ₄ and R ₅ together with the C atom to which they are connected form ring A, R ₆ , R ₇ , R ₈ are each independently hydrogen or R _b ; or R ₅ and R ₆ and the C atom to which they are connected form ring A. C atoms together form ring A, and R ₄ , R ₇ , and R ₈ are each independently hydrogen or R _b ; or R ₆ and R ₇ are the same or different R _b , or R ₆ and R ₇ are connected to the C they are connected to. The atoms together form Ring A, and R ₄ , R ₅ , and R ₈ are each independently hydrogen or R _b ; or R ₇ and R ₈ together with the C atom to which they are connected form Ring A, and R ₄ , R ₅ , and R ₆ are each independently hydrogen or R b; is independently hydrogen or R _b ; wherein any of the above R _b is independently halogen, hydroxyl, amino, cyano, unsubstituted or C ₁ -C ₆ alkyl substituted by R _a , unsubstituted or C ₁ -C ₆ alkoxy group substituted by R _a ; The ring A is a 3-8-membered cycloalkyl group that is unsubstituted or substituted by one or more R _a , or a 4-8 membered cycloalkyl group that is unsubstituted or substituted by R _a One-membered heterocycloalkyl; any of the above R _a is independently halogen, hydroxyl, amino, cyano, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl; when R _a is multiple , the R _a is the same or different; Option 3: In the heterocyclic compound represented by formula I, I, where 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 and R ₂ are each independently selected from: Halogen, hydroxyl, amino, cyano, unsubstituted or C ₁ -C ₆ alkyl substituted by one or more R _a , C ₁ -C ₆ alkoxy unsubstituted or substituted by one or more R _a , a 3-6-membered cycloalkyl group that is unsubstituted or substituted by one or more R _a ; When m is 2, 3, 4, the R ₁ is the same or different; When n is 2, 3, 4, the R 1 R ₂ is the same or different; R ₃ is hydrogen, unsubstituted or C ₁ -C ₆ alkyl substituted by one or more R _a ; R ₄ , R ₅ , R ₆ and R ₇ are each independently hydrogen or R _b , R ₈ is R _b ; or R ₄ , R ₅ , R ₆ , R ₇ and R ₈ are each independently hydrogen or R _b , and at least one of R ₄ , R ₅ , R ₆ , R ₇ and R ₈ is a cyano group; or R ₄ and R ₅ are the same or different R _b , or R ₄ and R ₅ together with the C atom to which they are connected form a ring A, and R ₆ , R ₇ , and R ₈ are each independently hydrogen or R _b ; or R ₅ and R ₆ together with the C atoms to which they are connected form ring A, and R ₄ , R ₇ , and R ₈ are each independently hydrogen or R _b ; or R ₆ and R ₇ are the same or different R _b , or R ₆ and R ₇ together with the C atom to which they are connected form ring A, R ₄ , R ₅ and R ₈ are each independently hydrogen or R _b ; or R ₇ and R ₈ and the C atom to which they are connected form ring A. Together, they form Ring A, and R ₄ , R ₅ , and R ₆ are each independently hydrogen or R _b ; wherein any of the above R _b is each independently halogen, hydroxyl, amino, cyano, unsubstituted, or R _aSubstituted C ₁ -C ₆ alkyl, unsubstituted or R _a substituted C ₁ -C ₆ alkoxy, unsubstituted or R _a substituted 3-6 membered cycloalkyl, unsubstituted or R _a Substituted 4-6 membered heterocycloalkyl; The ring A is a 3-8 membered cycloalkyl that is unsubstituted or substituted by one or more R _a , or a 4-8 membered heterocycle that is unsubstituted or substituted with R _a Alkyl group; Each of the above R _a is independently halogen, hydroxyl, amino group, cyano group, C ₁ -C ₆ alkyl group, C ₁ -C ₆ haloalkyl group; When R _a is multiple, the R _a is the same or different; Scheme 4: In the heterocyclic compound represented by formula I, I, where 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 and R ₂ are each independently selected from: Halogen, hydroxyl, amino, cyano, unsubstituted or C ₁ -C ₆ alkyl substituted by one or more R _a , C ₁ -C ₆ alkoxy unsubstituted or substituted by one or more R _a , a 3-6-membered cycloalkyl group that is unsubstituted or substituted by one or more R _a ; When m is 2, 3, 4, the R ₁ is the same or different; When n is 2, 3, 4, the R 1 R ₂ is the same or different; R ₃ is hydrogen, unsubstituted or C ₁ -C ₆ alkyl substituted by one or more R _a ; R ₄ , R ₅ , R ₆ and R ₇ are each independently hydrogen or R _b , R ₈ is R _b ; or R ₄ , R ₅ , R ₆ , R ₇ and R ₈ are each independently hydrogen or R _b , and at least one of R ₄ , R ₅ , R ₆ , R ₇ and R ₈ is a cyano group; or R ₄ and R ₅ are the same or different R _b , or R ₄ and R ₅ together with the C atom to which they are connected form a ring A, and R ₆ , R ₇ , and R ₈ are each independently hydrogen or R _b ; or R ₅ and R ₆ together with the C atoms to which they are connected form ring A, and R ₄ , R ₇ , and R ₈ are each independently hydrogen or R _b ; or R ₆ and R ₇ are the same or different R _b , or R ₆ and R ₇ together with the C atom to which they are connected form ring A, R ₄ , R ₅ and R ₈ are each independently hydrogen or R _b ; or R ₇ and R ₈ and the C atom to which they are connected form ring A. Together, they form Ring A, and R ₄ , R ₅ , and R ₆ are each independently hydrogen or R _b ; wherein any of the above R _b is each independently halogen, hydroxyl, amino, cyano, unsubstituted, or R _aSubstituted C ₁ -C ₆ alkyl, unsubstituted or R _a substituted C ₁ -C ₆ alkoxy, unsubstituted or R _a substituted 3-6 membered cycloalkyl, unsubstituted or R _a Substituted 4-6 membered heterocycloalkyl; The ring A is a 3-8 membered cycloalkyl that is unsubstituted or substituted by one or more R _a , or a 4-8 membered heterocycle that is unsubstituted or substituted with R _a Alkyl group; Each of the above R _a is independently deuterium, halogen, hydroxyl, amine group, cyano group, C ₁ -C ₆ alkyl group, C ₁ -C ₆ haloalkyl group; When R _a is multiple, The R _a are the same or different. The heterocyclic compound represented by formula I, its stereoisomer or a pharmaceutically acceptable salt thereof as described in claim 3, wherein the heterocyclic compound represented by formula I is any one of the following schemes: Scheme 1: The heterocyclic compound shown in formula I has structure II II, where m and n are respectively integers from 0 to 4; R ₁ and R ₂ are each independently selected from: halogen, hydroxyl, amino, cyano, C ₁ -C ₆ alkyl, 3-6 membered cycloalkyl group; the R ₁ or R ₂ is optionally substituted with a substituent selected from the following: halogen, hydroxyl, amino group, C ₁ -C ₆ alkyl group; when R ₁ is multiple, the R ₁ is the same or Different; when R ₂ is multiple, the R ₂ is the same or different; R ₃ is hydrogen, C ₁ -C ₆ alkyl, C ₁ -C ₆ deuterated alkyl or C ₁ -C ₆ haloalkyl; R _4. R ₅ , R ₆ and R ₇ are each independently hydrogen or C ₁ -C ₆ alkyl, and R ₈ is C ₁ -C ₆ alkyl; or R ₄ and R ₅ are formed together with the C atom to which they are connected. Ring A, R ₆ , R ₇ , R ₈ are each independently hydrogen or C ₁ -C ₆ alkyl; or R ₅ and R ₆ together with the C atom to which they are connected form Ring A, R ₄ , R ₇ , R ₈ is each independently hydrogen or C ₁ -C ₆ alkyl; or R ₆ and R ₇ together with the C atom to which they are connected form ring A, and R ₄ , R ₅ and R ₈ are each independently hydrogen or C ₁ - C ₆ alkyl; or R ₇ and R ₈ together with the C atoms to which they are connected form ring A, and R ₄ , R ₅ , and R ₆ are each independently hydrogen or C ₁ -C ₆ alkyl; 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 following: halogen, hydroxyl, amino, cyano, C ₁ -C ₆ alkyl , C ₁ -C ₆ haloalkyl; Option 2: The heterocyclic compound represented by formula I has structure II II, where m and n are respectively integers from 0 to 4; R ₁ and R ₂ are each independently selected from: halogen, hydroxyl, amino, cyano, C ₁ -C ₆ alkyl, 3-6 membered cycloalkyl group; the R ₁ or R ₂ is optionally substituted with a substituent selected from the following: halogen, hydroxyl, amino group, C ₁ -C ₆ alkyl group; when R ₁ is multiple, the R ₁ is the same or Different; when there are multiple R _2s , the R _2s are the same or different; R ₃ is hydrogen, C ₁ -C ₆ alkyl or C ₁ -C ₆ haloalkyl; R ₄ , R ₅ , R ₆ , R ₇ Each is independently hydrogen or C ₁ -C ₆ alkyl, R ₈ is C ₁ -C ₆ alkyl; or R ₄ and R ₅ together with the C atom to which they are connected form ring A, R ₆ , R ₇ , R ₈ is each independently hydrogen or C ₁ -C ₆ alkyl; or R ₅ and R ₆ together with the C atom to which they are connected form ring A, and R ₄ , R ₇ and R ₈ are each independently hydrogen or C ₁ - C ₆ alkyl; or R ₆ and R ₇ together with the C atoms to which they are connected form ring A, and R ₄ , R ₅ , and R ₈ are each independently hydrogen or C ₁ -C ₆ alkyl; or R ₇ and R ₈ together with the C atoms to which they are connected form Ring A, R ₄ , R ₅ and R ₆ are each independently hydrogen or C ₁ -C ₆ alkyl; the Ring A is a 3-8 membered cycloalkyl or 4- 8-membered heterocycloalkyl; The ring A is optionally substituted with a substituent selected from the following: halogen, hydroxyl, amino, cyano, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl. The heterocyclic compound represented by formula I, its stereoisomer or a pharmaceutically acceptable salt thereof as described in claim 3 or 4, wherein the heterocyclic compound represented by formula I has structure Ia or Ib Wherein, m, n, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ and R ₈ are as described in claim 3 or 4. The heterocyclic compound represented by formula I as described in claim 4, its stereoisomer or its pharmaceutically acceptable salt, wherein, Have structure , R ₁₁ is C ₁ -C ₃ alkyl, R ₁₂ is C ₁ -C ₃ haloalkyl; preferably, Have structure . The heterocyclic compound represented by formula I as described in claim 4, its stereoisomer or its pharmaceutically acceptable salt, wherein, Have structure , R ₂₁ , R ₂₂ , and R ₂₃ are the same or different halogens; preferably, the R ₂₁ , R ₂₂ , and R ₂₃ are independently F or Cl; more preferably, Have structure . The heterocyclic compound represented by formula I, its stereoisomer or a pharmaceutically acceptable salt thereof as described in claim 3 or 4, wherein R ₄ and R ₅ together with the C atom to which they are connected form ring A, R ₆ , R ₇ , and R ₈ are each independently hydrogen, hydroxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl; or R ₅ and R ₆ together with the C atom to which they are connected form Ring A, R ₄ , R ₇ , and R ₈ are each independently hydrogen, hydroxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl; or R ₆ and R ₇ together with the C atom to which they are connected form Ring A, R ₄ , R ₅ , and R ₈ are each independently hydrogen, hydroxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl; or R ₇ and R ₈ together with the C atom to which they are connected form Ring A, R ₄ , R ₅ , and R ₆ are each independently hydrogen, hydroxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl; preferably, R ₄ and R ₅ are formed together with the C atoms to which they are connected. Ring A, R ₆ , R ₇ , R ₈ are each independently hydrogen; or R ₅ and R ₆ together with the C atom to which they are connected form Ring A, R ₄ , R ₇ , R ₈ are hydrogen; or R ₆ and R ₇ and the C atoms to which they are connected together form ring A, and R ₄ , R ₅ and R ₈ are hydrogen; or R ₇ and R ₈ together with the C atoms to which they are connected form ring A, and R ₄ , R ₅ and R ₆ is hydrogen; The ring A is a 3-6 membered cycloalkyl group or a 4-8 membered heterocycloalkyl group; Preferably, the ring A is selected from: cyclopropane, cyclobutane, cyclopentane, cyclohexane Alkane; Preferably, the 4-8 membered heterocycloalkyl group has 1, 2 or 3 heteroatoms selected from N, O or S; when there are multiple heteroatoms, the heteroatoms are the same or different. The heterocyclic compound represented by formula I, its stereoisomer or its pharmaceutically acceptable salt as described in claim 3 or 4, wherein R ₄ , R ₅ , R ₆ and R ₇ are hydrogen, and R ₈ is C ₁ -C ₃ alkyl. The heterocyclic compound represented by formula I as described in claim 3 or 4, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein R ₄ and R ₅ are the same or different R _b , R ₆ , R _7. R ₈ are each independently hydrogen, hydroxyl, halogen, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl; or R ₆ and R ₇ are the same or different R _b , R ₄ , R ₅ , R ₈ is each independently hydrogen, hydroxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl; R _b is halogen, hydroxyl, cyano group, C ₁ -C ₆ alkyl; Preferably, R _b For fluorine, chlorine, hydroxyl, cyano, methyl, ethyl, propyl. The heterocyclic compound represented by formula I, its stereoisomer or a pharmaceutically acceptable salt thereof as described in claim 3, wherein the heterocyclic compound represented by formula I has the structure Ic, Id or Ie Ic. Id, Ie; wherein, rings K, m, n, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ and R ₈ are as described in claim 3. The heterocyclic compound represented by formula I, its stereoisomer or its pharmaceutically acceptable salt as described in claim 3 or 11, wherein ring K is a 5-membered or 6-membered N-containing heteroaromatic ring; preferably , Ring K is a pyridine ring, a pyrimidine ring, a pyrazine ring, and a pyridazine ring. The heterocyclic compound represented by formula I as described in any one of claims 3 to 4 and claim 8, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein R ₃ is a deuterated methyl. The heterocyclic compound represented by formula I, its stereoisomer or a pharmaceutically acceptable salt thereof as described in claim 1 or 2, wherein the heterocyclic compound represented by formula I satisfies the following conditions (1) to conditions One or more conditions in (21): (1) The m is 2 or 3, such as 2; (2) The n is 1 or 3, such as 3, (3) The ring K is 5 to 6 members Heteroaromatic ring. In the 5 to 6-membered heteroaromatic ring, the heteroatom is N and the number of heteroatoms is 1 or 2. For example, the ring K is pyridine; (4) Each R ₁ is independently halogen, C ₁ -C ₆ alkyl or 3-6 membered cycloalkyl that is unsubstituted or substituted by one or more R _a . Preferably, each R ₁ is independently halogen, unsubstituted or substituted by one or more R _a Substituted C ₁ -C ₆ alkyl, for example, each R ₁ is independently halogen, trifluoromethyl, methyl or trideuterated methyl, and for example, each R ₁ is independently unsubstituted or substituted by a or multiple R _a substituted C ₁ -C ₆ alkyl groups; (5) When m is 2, each R ₁ is located at the ortho or para position of the heteroatom in ring K; (6) When m is 3, each R 1 R ₁ is adjacent and located in the ortho, meta or para position of the heteroatom in ring K; (7) In each R ₁ , each R _a is independently halogen or deuterium, for example, in each R ₁ , each R _a Each is independently a halogen; (8) R ₂ is each independently a halogen; (9) R ₃ is a C ₁ -C ₆ alkyl group that is unsubstituted or substituted by one or more R _a , for example, R ₃ is C ₁ -C ₆ alkyl, and for example, R ₃ is a C ₁ -C ₆ alkyl group substituted by one or more R _a ; (10) In R ₃ , each R _a is independently deuterium; (11) Each R _b Each is independently halogen, hydroxyl, cyano, 3-6 membered cycloalkyl or C ₁ -C ₆ alkyl that is unsubstituted or substituted by one or more R _a . Preferably, each R _b is independently halogen. , hydroxyl or C ₁ -C ₆ alkyl, for example, each R _b is independently a hydroxyl group or a C ₁ -C ₆ alkyl group; (12) In each R _b , each R _a is independently a halogen; (13) Each ring A is independently a 3-8 membered cycloalkyl group; (14) R ₄ , R ₅ , R ₆ and R ₇ are each independently hydrogen or R _b , and R ₈ is a C ₁ -C ₆ alkyl group; ( 15) R ₅ , R ₆ , R ₇ and R ₈ are each independently hydrogen or R _b , and R ₄ is cyano; (16) R ₄ , R ₅ , R ₇ and R ₈ are each independently hydrogen or R _b , and R ₆ is cyano; (17) R ₄ and R ₅ are each independently R _b , R ₆ , R ₇ and R ₈ are each independently R _b or hydrogen, for example, R ₄ and R ₅ are each independently R b is R _b , R ₆ and R ₇ are each independently hydrogen, C ₁ -C ₆ alkyl or hydroxyl, R ₈ is hydrogen, preferably, R ₄ and R ₅ are each independently R _b , R ₆ and R ₇ is each independently hydrogen or hydroxyl, R ₈ is hydrogen; (18) R ₆ and R ₇ are each independently R _b , R ₄ , R ₅ and R ₈ are each independently R _b or hydrogen, for example, R ₆ and R ₇ are each independently R _b , R ₄ and R ₅ are each independently hydrogen, C ₁ -C ₆ alkyl or hydroxyl, R ₈ is hydrogen; (19) R ₄ and R ₅ and the C to which they are connected The atoms together form Ring A, R ₆ and R ₇ are each independently hydrogen or hydroxyl, R ₈ is hydrogen, for example, R ₄ and R ₅ together with the C atom to which they are attached form Ring A, R ₆ and R ₇ are each independently Ground is hydrogen, R ₈ is hydrogen; (20) R ₅ and R ₆ form ring A together with the C atom to which they are connected, R ₄ and R ₇ are each independently hydrogen or hydroxyl, and R ₈ is hydrogen; (21) R ₆ and R ₇ together with the C atom to which they are connected form ring A, R ₄ and R ₅ are each independently hydrogen or hydroxyl, and R ₈ is hydrogen. The heterocyclic compound represented by formula I, its stereoisomer or a pharmaceutically acceptable salt thereof as described in claim 1, wherein the heterocyclic compound represented by formula I is any one of the following schemes: Scheme 1: m is 0, 1, 2, 3 or 4; n is 0, 1, 2, 3 or 4; Ring K is a 5 to 6-membered heteroaromatic ring, and in the 5 to 6-membered heteroaromatic ring, the heteroatom is N, The number of heteroatoms is 1 or 2; each R ₁ is independently halogen, unsubstituted or C ₁ -C ₆ alkyl or 3-6 membered cycloalkyl substituted by one or more R _a ; in each R ₁ , each R _a is independently halogen or deuterium; R ₂ is each independently halogen; R ₃ is a C ₁ -C ₆ alkyl group that is unsubstituted or substituted by one or more R _a ; in R ₃ , each R _a Each is independently deuterium; R ₄ , R ₅ , R ₆ and R ₇ are each independently hydrogen or R _b , R ₈ is C ₁ -C ₆ alkyl; or, R ₅ , R ₆ , R ₇ and R ₈ Each is independently hydrogen or R _b , and R ₄ is cyano; or, R ₄ , R ₅ , R ₇ and R ₈ are each independently hydrogen or R _b , and R ₆ is cyano; or R ₄ and R ₅ is each independently R _b , R ₆ and R ₇ are each independently hydrogen, C ₁ -C ₆ alkyl or hydroxyl, R ₈ is hydrogen; or, R ₆ and R ₇ are each independently R _b , R ₄ and R ₅ are each independently hydrogen, C ₁ -C ₆ alkyl or hydroxyl, and R ₈ is hydrogen; or, R ₄ and R ₅ together with the C atom to which they are connected form ring A, and R ₆ and R ₇ are each independently are independently hydrogen or hydroxyl, R ₈ is hydrogen; or, R ₅ and R ₆ together with the C atom to which they are attached form Ring A, R ₄ and R ₇ are each independently hydrogen or hydroxyl, and R ₈ is hydrogen; or , R ₆ and R ₇ together with the C atoms to which they are connected form ring A, R ₄ and R ₅ are each independently hydrogen or hydroxyl, R ₈ is hydrogen; each R _b is independently halogen, hydroxyl, cyano, 3-6 membered cyclic alkyl or C ₁ -C ₆ alkyl that is unsubstituted or substituted by one or more R _a ; In each R _b , each R _a is independently halogen; each ring A is independently 3 -8-membered cycloalkyl; Scheme 2: m is 2 or 3; n is 3; Ring K is pyridine; each R ₁ is independently halogen, unsubstituted or C ₁ -C substituted by one or more R _{a 6} alkyl; when m is 2, each R ₁ is located in the ortho or meta position of the heteroatom in ring K; when m is 3, each R ₁ is adjacent and located in the ortho or meta position of the heteroatom in ring K Or para position; In each R ₁ , each R _a is independently halogen or deuterium; R ₂ is each independently halogen; R ₃ is a C ₁ -C ₆ alkyl group that is unsubstituted or substituted by one or more R _a ; In R ₃ , each R _a is independently deuterium; R ₄ and R ₅ are each independently R _b , R ₆ and R ₇ are each independently hydrogen or hydroxyl, and R ₈ is hydrogen; or, R ₄ and R ₅ together with the C atoms to which they are connected form ring A, R ₆ and R ₇ are each independently hydrogen, and R ₈ is hydrogen; each R _b is independently halogen, hydroxyl or C ₁ -C ₆ alkyl; each ring A is each independently a 3-8 membered cycloalkyl group; Scheme 3: m is 2 or 3; n is 3; Ring K is pyridine; each R ₁ is independently halogen, trifluoromethyl, methyl or trideuterium Substitute methyl; when m is 2, each R ₁ is located in the ortho or para position of the heteroatom in ring K; when m is 3, each R ₁ is adjacent and located in the ortho or meta position of the heteroatom in ring K or para position; R ₂ is each independently halogen; R ₃ is a C ₁ -C ₆ alkyl group that is unsubstituted or substituted by one or more R _a ; in R ₃ , each R _a is independently deuterium; R ₄ and R ₅ are each independently R _b , R ₆ and R ₇ are each independently hydrogen or hydroxyl, and R ₈ is hydrogen; or, R ₄ and R ₅ together with the C atom to which they are connected form ring A, R ₆ and R ₇ is each independently hydrogen, and R ₈ is hydrogen; each R _b is independently hydroxyl or C ₁ -C ₆ alkyl; each ring A is independently 3-8 membered cycloalkyl; Scheme 4: m is 2; n is 3; Ring K is pyridine; each R ₁ is independently a C ₁ -C ₆ alkyl group that is unsubstituted or substituted by one or more R _a ; when m is 2, each R ₁ is located in ring K Ortho or para position of the heteroatom in the medium; In each R ₁ , each R _a is independently a halogen; R ₂ is each independently a halogen; R ₃ is a C ₁ -C ₆ alkyl group; R ₄ and R ₅ are each independently Ground is R _b , R ₆ and R ₇ are each independently hydrogen or hydroxyl, and R ₈ is hydrogen; or, R ₄ and R ₅ form ring A together with the C atom to which they are connected, and R ₆ and R ₇ are each independently is hydrogen, R ₈ is hydrogen; each R _b is independently a hydroxyl group or a C ₁ -C ₆ alkyl group; each ring A is independently a 3-8 membered cycloalkyl group; Scheme 5: m is 2 or 3; n is 3; Ring K is pyridine; each R ₁ is independently halogen, trifluoromethyl, methyl or trideuterated methyl; when m is 2, each R ₁ is located in the ortho position of the heteroatom in ring K or Para position, when m is 3, each R ₁ is adjacent and located in the ortho position, meta position or para position of the heteroatom in ring K; in each R ₁ , each R _a is independently halogen or deuterium; R ₂ is each is independently halogen; R ₃ is a C ₁ -C ₆ alkyl group substituted by one or more _Ra ; in R ₃ , each R _a is independently deuterium; R ₄ and R ₅ are each independently R _b , R ₆ and R ₇ are each independently hydrogen or hydroxyl, and R ₈ is hydrogen; each R _b is independently hydroxyl or C ₁ -C ₆ alkyl; each ring A is independently a 3-8-membered cycloalkyl. The heterocyclic compound represented by Formula I, its stereoisomer or a pharmaceutically acceptable salt thereof as described in claim 1, wherein the heterocyclic compound represented by Formula I satisfies the following conditions (1) to (11) ) one or more conditions in: (1) Ring K is pyridine or pyrimidine; (2) R ₁ is methyl, trifluoromethyl, difluoromethyl, cyclopropyl, fluorine or trideuterated methyl; ( 3) R ₂ is fluorine or chlorine; (4) R ₃ is methyl or trideuterated methyl; (5) R ₄ and R ₅ are each independently hydrogen, fluorine, hydroxyl, methyl, cyclopropyl, trideuteryl Fluoromethyl, cyano or difluoromethyl; Alternatively, R ₄ and R ₅ together with the C atom to which they are connected form ring A, and the ring A is a cyclopropyl group: Alternatively, R ₅ and R ₆ and the C atom to which they are connected form a ring A. The connected C atoms together form ring A, which is cyclopropyl; (6) R ₆ and R ₇ are each independently hydrogen, hydroxyl, methyl or cyano; (7) R ₈ is hydrogen or methyl ; (8) for , , , , , , or ; (9) for , , , , , , , , , , , , , , or ; (10) for or ; (11) for , , or . The heterocyclic compound represented by formula I, its stereoisomer or a pharmaceutically acceptable salt thereof as described in claim 1, wherein the heterocyclic compound represented by formula I is any of the following compounds: . The heterocyclic compound represented by formula I, its stereoisomer or a pharmaceutically acceptable salt thereof as described in claim 1, wherein the heterocyclic compound represented by formula I is any of the following compounds: . A pharmaceutical composition, which includes a heterocyclic compound represented by formula I in any one of claims 1 to 18, its stereoisomer or a pharmaceutically acceptable salt thereof, and optionally i) one or more active drugs; and/or ii) Pharmaceutically acceptable carrier. A use of the heterocyclic compound represented by formula I in any one of claims 1 to 18, its stereoisomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described in claim 19, which use includes: 1) Inhibit Polθ activity; 2) Prevent and treat Polθ-mediated diseases; 3) Preparation of drugs, pharmaceutical compositions or preparations for inhibiting Polθ activity; 4) Preparation of drugs, pharmaceutical compositions or preparations for preventing and treating Polθ-mediated diseases; Preferably, the Polθ-mediated disease is 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.