WO2024022519A1 - Heterocycle fused pyrimidine compound and use thereof - Google Patents

Abstract

Provided are a compound of formula (I) or a pharmaceutically acceptable salt thereof as a USP1 inhibitor and a pharmaceutical composition comprising the compound or the pharmaceutically acceptable salt thereof. The compound and the pharmaceutical composition can be used for preventing or treating diseases related to USP1.

Description

Heterocyclic pyrimidine compounds and their applications

This disclosure claims the priority of the earlier application submitted to the State Intellectual Property Office of China on July 28, 2022, with the patent application number 202310900509.6 and the invention title "Heterocyclopyrimidine Compounds and Their Applications". The entire contents of the above-mentioned prior applications are incorporated into this disclosure by reference.

Technical field

The present disclosure belongs to the field of medical technology and relates to a class of heterocyclic pyrimidine compounds, or pharmaceutically acceptable salts thereof, pharmaceutical compositions containing them, and as ubiquitin-specific protease 1 (USP1) inhibitors in the prevention or treatment of Use in USP1-related diseases.

Background technique

Ubiquitination is a reversible process that involves a series of deubiquitinating enzymes (DUBs) that regulate a variety of cellular processes by deubiquitinating substrates. DUBs are encoded by approximately 100 human genes and are divided into six families, the largest of which is the ubiquitin-specific proteases (USPs) with more than 50 members. DUBs and their substrate proteins are often dysregulated in cancer, which supports the idea that targeting specific DUB enzymes can participate in tumor growth, survival, differentiation, and maintenance of the tumor microenvironment by improving ubiquitination and degradation of oncogenic substrates and regulating them. hypothesis of the activity of other key proteins (Hussain, S., et.al., "DUBs and cancer: The role of deubiquitinatingenzymes as oncogenes, non-oncogenes and tumor suppressors." Cell Cycle 8, 1688-1697 (2009)) .

Ubiquitin-specific protease 1 (USP1) is a cysteine isopeptidase of the USP subfamily of DUBs. Full-length human USP1 consists of 785 amino acids, including a catalytic triad consisting of Cys90, His593 and Asp751 (Nijman, S.M.B., et al. "The deubiquitinating enzyme USPl regulates the fanconi anemia pathway.Mal.Cell 17,331-339 (2005)). USP1 plays a role in DNA damage repair. USP1 itself is relatively inactive, and full enzymatic activity can only be obtained by combining with the cofactor UAF1 to form a complex required for deubiquitinating enzyme activity. USP1/UAFl complex Deubiquitinated monoubiquitinated PCNA (proliferating cell nuclear antigen) and monoubiquitinated FANCD2 (Fanconi anemia group complementary group D2), these two proteins are involved in translation synthesis (TLS) and Fanconi anemia ( FA) pathway. These two pathways are required to repair DNA damage caused by DNA cross-linking agents such as cisplatin and mitomycin C (MMC). The USPl/UAFl complex also deubiquitinates FANCI( Fanconi anemia complementation group I). The importance of these findings was further confirmed by experiments showing that mice lacking USP1 are highly sensitive to DNA damage. Interestingly, USP1 expression is significantly increased in many cancers. Blocking USP1 inhibits DNA repair , can induce apoptosis in multiple myeloma cells and can also enhance the sensitivity of lung cancer cells to cisplatin. These indicate that USP1 is a promising target for chemotherapy in certain cancers.

Taken together, targeted inhibition of the USP1 protein is a potential way to prevent or treat cancer and other diseases. Therefore, the development of small molecule inhibitors of USP1 is necessary.

Contents of the invention

In one aspect, the present disclosure relates to a compound of formula (I), or a pharmaceutically acceptable salt thereof,

in, Represents a single or double bond;

X is selected from N, CR ³ , NR ³ or C(O);

Y is selected from NR ⁴ or C(O);

Z is selected from CR ⁵ or NR ⁵ ;

R ³ , R ⁴ , R ⁵ are independently selected from H, halogen, CN, OH, NH ₂ , -C(O)OR ^x , -C(O)R ^x , -OC ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₁₀ cycloalkyl or 4-7 membered heterocyclyl, the NH ₂ , -OC ₁ -C ₆ Alkyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₁₀ cycloalkyl or 4-7 membered heterocyclyl is optionally substituted by R ^x ;

Ring A is selected from C _{6 -C 10} aryl or _5-10 membered heteroaryl, which is optionally substituted by one or more R ^a ;

Ring B is selected from C ₆ -C ₁₀ aryl, 5-10 membered heteroaryl, 4-10 membered heterocyclyl, C ₄ -C ₁₀ cycloalkenyl or C ₃ -C ₁₀ cycloalkyl, the C ₆ -C ₁₀ aryl, 5-10 membered heteroaryl, 4-10 membered heterocyclyl, C ₄ -C ₁₀ cycloalkenyl or C ₃ -C ₁₀ cycloalkyl optionally substituted by one or more R ^b ;

Ring C is selected from C ₆ -C ₁₀ aryl _{or 5-10} membered heteroaryl, which is optionally substituted by one or more R ^c ;

Each R ^a , R ^b , R ^c is independently selected from halogen, CN, OH, NH ₂ , -C(O)OR ^x , C ₁ -C ₆ alkyl, -OC ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl or 4-10 membered heterocyclyl, the NH ₂ , C ₁ -C ₆ alkyl, -OC ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl or 4-10 membered Heterocyclyl is optionally substituted with ^Rx ,

Or R ^b , R ^c and the atoms to which they are connected together form a C ₄ -C ₁₀ cycloalkenyl group or a 4-10 membered heterocyclyl group, and the C ₄ -C ₁₀ cycloalkenyl group or 4-10 membered heterocyclyl group can be any The chosen land is replaced by R ^x ;

R ¹ and R ² are independently selected from H, halogen, CN, OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl or 4-10 membered heterocyclyl, the OH, NH _2. C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl or 4-10 membered heterocyclyl is optionally substituted by R ^x ,

Or R ¹ , R ² and the atoms to which they are connected together form a C ₃ -C ₁₀ cycloalkyl group or a 4-7 membered heterocyclyl group, and the C ₃ -C ₁₀ cycloalkyl group or 4-7 membered heterocyclyl group can be any The chosen land is replaced by R ^x ;

^R _{_ _ _ _ _ _ _ _ _} C ₁₀ cycloalkyl or 4-7 membered heterocyclyl substituted.

In some embodiments, R ³ , R ⁴ , R ⁵ are independently selected from H, -C(O)OR ^x , -C(O)R ^x , -OC ₁ -C ₆ alkyl, C ₁ -C ₆ Alkyl, C ₃ -C ₁₀ cycloalkyl or 4-7 membered heterocyclyl, the -OC ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl or 4- The 7-membered heterocyclyl is optionally substituted with ^Rx .

In some embodiments, R ³ , R ⁴ , and R ⁵ are independently selected from H, C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl, or 4-7 membered heterocyclyl, and the C ₁ - C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl or 4-7 membered heterocyclyl is optionally substituted by R ^x .

In some embodiments, R ³ , R ⁴ , R ⁵ are independently selected from H, CH ₃ , CH ₂ CH ₃ , cyclopropyl, oxetanyl or azetidinyl, said CH ₃ , CH _2CH3 , _cyclopropyl , oxetanyl or azetidinyl is optionally substituted by ^Rx .

In some embodiments, R ³ , R ⁴ , R ⁵ are independently selected from H, CH ₃ , CH ₂ CH ₃ , CH ₂ CH ₂ N(CH ₃ ) ₂ , cyclopropyl, oxetanyl, or

In some embodiments, R ⁴ is selected from H, C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl, or 4-7 membered heterocyclyl, said C ₁ -C ₆ alkyl, C ₃ - C ₁₀ cycloalkyl or 4-7 membered heterocyclyl is optionally substituted by ^Rx .

In some embodiments, R ⁴ is selected from H, CH ₃ , CH ₂ CH ₃ , cyclopropyl, oxetanyl, azetidinyl, tetrahydrofuryl, or tetrahydropyrrolyl, said CH ₃ , CH _2CH3 , _cyclopropyl , oxetanyl, azetidinyl, tetrahydrofuryl or tetrahydropyrrolyl is optionally substituted by ^Rx .

In some embodiments, ^R4 _is selected from H _{, CH3 , CH2CH3} , _CHF2 , _CH2CH2F , _{CH2CHF2 , CH2CF3 , CH2CH2N} ( _CH3 ) ₂ , cyclopropyl, oxetanyl,

In some embodiments, ^R5 is selected from H or _Ci - _C6 alkyl.

In some embodiments, ^R5 is selected from H or _CH3 .

In some embodiments, Ring A is selected from phenyl or 5-6 membered heteroaryl, which is optionally substituted with one or more ^Ra .

In some embodiments, Ring A is selected from phenyl, pyridyl, or pyrimidinyl, which is optionally substituted with one or more ^Ra .

In some embodiments, Ring B is selected from C ₆ -C ₁₀ aryl or _5-10 membered heteroaryl optionally substituted by _one or more Replaced by R ^b .

In some embodiments, Ring B is selected _from C ₆ -C ₁₀ aryl optionally substituted with one or more _R ^b .

In some embodiments, Ring B is selected from phenyl, optionally substituted with one or more ^Rb .

In some embodiments, Ring C is selected from 5-10 membered heteroaryl groups optionally substituted with one or more ^Rc .

In some embodiments, Ring C is selected from 5-6 membered heteroaryl groups optionally substituted with one or more ^Rc .

In some embodiments, Ring C is selected from pyrazolyl or imidazolyl, which is optionally substituted with one or more ^Rc .

In some embodiments, each ^Ra , ^Rb , ^Rc is independently selected from halogen, C ₁ -C ₆ alkyl, -OC ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl, or 4- 10-membered heterocyclyl group, the C ₁ -C ₆ alkyl group, -OC ₁ -C ₆ alkyl group, C ₃ -C ₁₀ cycloalkyl group or 4-10 membered heterocyclyl group is optionally substituted by R ^x .

In some embodiments, each ^Ra , ^Rb , ^Rc is independently selected from halogen, C ₁ -C ₆ alkyl, -OC ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl, or 4- 6-membered heterocyclyl, the C ₁ -C ₆ alkyl, -OC ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl or 4-6 membered heterocyclyl is optionally substituted by R ^x .

In some embodiments, each ^Ra , ^Rb , ^Rc is independently selected from halogen, _CH3 , -CH( _CH3 ) ₂ , _{-CH2CH3 ,} -O- _CH3 , cyclopropyl, oxygen Heterocyclidyl or azetidinyl, the CH ₃ , -CH(CH ₃ ) ₂ , -CH ₂ CH ₃ , -O-CH ₃ , cyclopropyl, oxetanyl or azetidinyl The group is optionally substituted by ^Rx .

In some embodiments ^, each Ra, ^Rb , ^Rc is independently selected from F, _CH3 , -CH( _CH3 ) ₂ , _{-CH2CH3 , -CHF2} , _-CF3 , -O- CH ₃ , -O-CHF ₂ , cyclopropyl, oxetanyl or

In some embodiments, R ^a is selected from C ₁ -C ₆ alkyl, -OC ₁ -C ₆ alkyl, or C ₃ -C ₁₀ cycloalkyl, said C ₁ -C ₆ alkyl, OC ₁ -C ₆ alkyl or C ₃ -C ₁₀ cycloalkyl is optionally substituted by R ^x .

In some embodiments, Ra ^is selected from isopropyl, -O- _CH3 , -O- _CHF2 , or cyclopropyl.

In some embodiments, R ^b is selected from halogen or -OC ₁ -C ₆ alkyl.

In some embodiments, ^Rb is selected from F or -O- _CH3 .

In some embodiments, ^Rc is selected from C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl, or 4-10 membered heterocyclyl, said C ₁ -C ₆ alkyl, C ₃ -C ₁₀ Cycloalkyl or 4-10 membered heterocyclyl is optionally substituted by ^Rx .

In some embodiments, R ^c is selected from C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, or 4-6 membered heterocyclyl, said C ₁ -C ₆ alkyl, C ₃ -C ₆ Cycloalkyl or 4-6 membered heterocyclyl is optionally substituted by ^Rx .

In some embodiments, ^Rc is selected from _CH3 , -CH( _CH3 ) ₂ , _-CH2CH3 , _-CF3 , _cyclopropyl , oxetanyl, or

In some embodiments, R ^b , R ^c and the atoms to which they are attached together form a 4-10 membered heterocyclyl group that is optionally substituted with R ^x .

In some embodiments, R ¹ and R ² are independently selected from H, halogen, C ₁ -C ₆ alkyl or C ₃ -C ₁₀ cycloalkyl, said C ₁ -C ₆ alkyl or C ₃ -C ₁₀ Cycloalkyl is optionally substituted with ^Rx .

In some embodiments, R ¹ and R ² are both H.

In some embodiments, ^Rx is selected _from halogen, _{NH2 ,} or _C1 - _C6 alkyl optionally replaced by _{C1 - C6alkyl} , _C3- C ₁₀ cycloalkyl or 4-7 membered heterocyclyl substituted.

In some embodiments, ^Rx is selected from halogen, _NH2 , or _Ci - _C6 alkyl, with the _NH2 optionally substituted by Ci _{- C6} alkyl.

In some embodiments, ^Rx is selected from F, N( _CH3 ) ₂ , or _Ci - _C6 alkyl.

In some embodiments, the compound represented by formula (I) or a pharmaceutically acceptable salt thereof is selected from the group consisting of compounds represented by formula (II) or a pharmaceutically acceptable salt thereof,

Ring A, Ring B, Ring C, R ¹ , R ² , R ⁴ and R ⁵ are as defined above.

In some embodiments, the compound represented by formula (I) or a pharmaceutically acceptable salt thereof is selected from the group consisting of compounds represented by formula (III) or a pharmaceutically acceptable salt thereof,

in, Represents a single bond or a double bond; Ring A, Ring B, Ring C, R ¹ , R ² , X and Z are as defined above.

In some embodiments, the compound represented by formula (III) or a pharmaceutically acceptable salt thereof is selected from the group consisting of compounds represented by formula (III-a) or a pharmaceutically acceptable salt thereof,

Ring A, Ring B, Ring C, R ¹ , R ² and R ⁵ are as defined above.

In some embodiments, the compound represented by formula (III) or a pharmaceutically acceptable salt thereof is selected from the group consisting of compounds represented by formula (III-b) or a pharmaceutically acceptable salt thereof,

Ring A, Ring B, Ring C, R ¹ , R ² , R ³ and R ⁵ are as defined above.

In some embodiments, the compound represented by formula (III) or a pharmaceutically acceptable salt thereof is selected from the group consisting of compounds represented by formula (III-c) or a pharmaceutically acceptable salt thereof,

Ring A, Ring B, Ring C, R ¹ , R ² , R ³ and R ⁵ are as defined above.

In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof is selected from the following compounds or a pharmaceutically acceptable salt thereof,

In another aspect, the present disclosure provides a pharmaceutical composition comprising a compound of formula (I) of the present disclosure or a pharmaceutically acceptable salt thereof and pharmaceutically acceptable excipients.

On the other hand, the present disclosure provides a method for treating a disease mediated by USP1 in a mammal, comprising administering to a mammal in need of the treatment, preferably a human, a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, or pharmaceutical compositions thereof.

On the other hand, the present disclosure provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in the preparation of a medicament for preventing or treating USP1-mediated diseases.

In another aspect, the present disclosure provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in preventing or treating USP1-mediated diseases.

In another aspect, the present disclosure provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof for preventing or treating USP1-mediated diseases.

In some embodiments, the USP1-mediated disease is neoplasia.

In some embodiments, the tumor is, for example, a solid tumor, adenocarcinoma, or hematologic cancer.

Definitions and explanations of terms

Unless otherwise stated, the terms used in this disclosure have the following meanings. The definitions of groups and terms recorded in this disclosure include their definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, and examples. The definitions of specific compounds, etc., can be arbitrarily combined and combined with each other. A particular term should not be considered uncertain or unclear in the absence of a specific definition, but should be understood in accordance with its ordinary meaning in the art. Where a trade name appears herein, it is intended to refer to its corresponding trade name or its active ingredient.

in this article Indicates the connection site.

The schematic representation of racemic or enantiopure compounds herein is taken from Maehr, J. Chem. Ed. 1985, 62: 114-120. Unless otherwise stated, wedge and virtual wedge bonds are used Represents the absolute configuration of a stereocenter, using black real and imaginary bonds. Represents the relative configuration of a stereocenter (such as the cis-trans configuration of an alicyclic compound).

The term "tautomer" refers to a functional group isomer resulting from the rapid movement of an atom in a molecule between two positions. Compounds of the present disclosure may exhibit tautomerism. Tautomeric compounds can exist in two or more interconvertible species. Tautomers generally exist in equilibrium, and attempts to isolate a single tautomer usually yield a mixture whose physical and chemical properties are consistent with the mixture of compounds. The position of equilibrium depends on the chemical properties within the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the keto form is dominant; in phenols, the enol form is dominant. This disclosure encompasses all tautomeric forms of the compounds.

The term "stereoisomer" refers to isomers resulting from different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers and diastereomers.

The compounds of the present disclosure may have asymmetric atoms such as carbon atoms, sulfur atoms, nitrogen atoms, phosphorus atoms, or asymmetric double bonds, and therefore the compounds of the present disclosure may exist in specific geometric or stereoisomeric forms. Specific geometric or stereoisomeric forms may be cis and trans isomers, E and Z geometric isomers, (-)- and (+)-enantiomers, (R)- and (S) )-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic or other mixtures thereof, such as enantiomers or diastereomers Enriched mixtures, all of the above isomers and mixtures thereof are within the definition of the compounds of the present disclosure. There may be additional asymmetric carbon atoms, asymmetric sulfur atoms, asymmetric nitrogen atoms or asymmetric phosphorus atoms in substituents such as alkyl groups. These isomers and their mixtures involved in all substituents are also included in Within the definition of the compounds of the present disclosure. The compounds of the present disclosure containing asymmetric atoms can be isolated in an optically active pure form or in a racemic form. The optically active pure form can be resolved from a racemic mixture or synthesized by using chiral starting materials or chiral reagents. .

The term "substituted" means that any one or more hydrogen atoms on a specific atom are replaced by a substituent, as long as the valence state of the specific atom is normal and the substituted compound is stable. When the substituent is oxo (i.e. =O), it means that two hydrogen atoms are replaced, and oxo does not occur on aromatic groups.

The term "optionally" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes both the occurrence and absence of the stated event or circumstance. For example, the ethyl group is "optionally" substituted by halogen, which means that the ethyl group can be unsubstituted (CH ₂ CH ₃ ), monosubstituted (CH ₂ CH ₂ F, CH ₂ CH ₂ Cl, etc.), or polysubstituted. (CHFCH ₂ F, CH ₂ CHF ₂ , CHFCH ₂ Cl, CH ₂ CHCl _2, etc.) or completely substituted (CF ₂ CF ₃ , CF ₂ CCl ₃ , CCl ₂ CCl _3, etc.). It will be understood by those skilled in the art that any substitution or substitution pattern that is sterically impossible and/or cannot be synthesized will not be introduced for any group containing one or more substituents.

When any variable (eg, R ^a , R ^b ) occurs more than once in the composition or structure of a compound, its definition in each instance is independent. For example, if a group is replaced by 2 R ^b , there are separate options for each R ^b .

When the number of a linking group is 0, such as -(CH ₂ ) ₀ -, it means that the linking group is a bond.

When one of the variables is selected from a chemical bond or is absent, it means that the two groups it is connected to are directly connected. For example, when L in A-L-Z represents a bond, it means that the structure is actually A-Z.

When the linking group mentioned in this article does not specify the direction of connection, the direction of connection is arbitrary. For example, when the structural unit When L ¹ is selected from "C ₁ -C ₃ alkylene-O", then L ¹ can connect ring Q and R ¹ in the direction from left to right to form "ring QC ₁ -C ₃ alkylene Group -OR ¹ ”, you can also connect ring Q and R ¹ from right to left to form “ring QOC ₁ -C ₃ alkylene group -R ¹ ”.

When a substituent's bond is cross-linked to two atoms on a ring, the substituent can be bonded to any atom on the ring. For example, structural unit Indicates that R ⁵ can be substituted at any position on the benzene ring.

In this article, bonds depicted by solid and dashed lines Represents a single or double bond. For example, structural unit Include

_Cm - _Cn as used herein refers to having an integer number of carbon atoms in the range of mn. For example, "C ₁ -C ₁₀ " means that the group can have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, 9 carbon atoms or 10 carbon atoms.

The term "alkyl" refers to a hydrocarbon group of the general formula C _n H _2n+1 , which alkyl group may be straight or branched. The term "C ₁ -C ₁₀ alkyl" is understood to mean a straight-chain or branched saturated hydrocarbon radical having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. Specific examples of the alkyl group include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2- Methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-di Methylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, etc.; the term "C ₁ -C ₆ alkyl "can be understood to mean an alkyl group with 1 to 6 carbon atoms. Specific examples include but are not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec. Butyl, tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc. The term "C ₁ -C ₃ alkyl" is understood to mean a straight-chain or branched saturated alkyl group having 1 to 3 carbon atoms. The "C ₁ -C ₁₀ alkyl" may include "C ₁ -C ₆ alkyl" or "C ₁ -C ₃ alkyl" and other ranges, and the "C ₁ -C ₆ alkyl" may further include " C ₁ -C ₃ alkyl”.

The term "alkenyl" refers to a linear or branched unsaturated lipid composed of carbon atoms and hydrogen atoms and having at least one double bond. Aliphatic hydrocarbon group. The term "C ₂ -C ₁₀ alkenyl" is understood to mean a straight-chain or branched unsaturated hydrocarbon radical containing one or more double bonds and having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, "C ₂ -C ₁₀ alkenyl" is preferably "C ₂ -C ₆ alkenyl", more preferably "C ₂ -C ₄ alkenyl", and even more preferably C ₂ or C ₃ alkenyl. It will be appreciated that where the alkenyl group contains more than one double bond, the double bonds may be separated or conjugated to each other. Specific examples of the alkenyl group include, but are not limited to, vinyl, allyl, (E)-2-methylvinyl, (Z)-2-methylvinyl, (E)-but-2-enyl , (Z)-but-2-enyl, (E)-but-1-enyl, (Z)-but-1-enyl, isopropenyl, 2-methylprop-2-enyl, 1 -Methylprop-2-enyl, 2-methylprop-1-enyl, (E)-1-methylprop-1-enyl or (Z)-1-methylprop-1-enyl wait.

The term "alkynyl" refers to a linear or branched unsaturated aliphatic hydrocarbon group composed of carbon atoms and hydrogen atoms and having at least one triple bond. The term "C ₂ -C ₁₀ alkynyl" is understood to mean a linear or branched unsaturated hydrocarbon radical containing one or more triple bonds and having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. Examples of "C ₂ -C ₁₀ alkynyl" include, but are not limited to, ethynyl (-C≡CH), propynyl (-C≡CCH _3, -CH ₂ C≡CH), but-1-ynyl, butyl -2-alkynyl or but-3-ynyl. "C ₂ -C ₁₀ alkynyl" may include "C ₂ -C ₆ alkynyl" and "C ₂ -C ₃ alkynyl", and examples of "C ₂ -C ₃ alkynyl" include ethynyl (-C≡CH) , prop-1-ynyl (-C≡CCH ₃ ), prop-2-ynyl (-CH ₂ C≡CH).

The term "cycloalkyl" refers to a fully saturated carbocyclic ring that exists in the form of a single ring, a branched ring, a bridged ring or a spiro ring. Unless otherwise indicated, the carbocyclic ring is generally 3 to 10 membered. The term "C ₃ -C ₁₀ cycloalkyl" is understood to mean a saturated monocyclic, paracyclic, spirocyclic or bridged ring having 3 to 10 carbon atoms. Specific examples of the cycloalkyl group include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, norbornyl (bicyclo[2.2 .1]heptyl), bicyclo[2.2.2]octyl, adamantyl, spiro[4.5]decyl, etc. The term "C ₃ -C ₁₀ cycloalkyl" may include "C ₃ -C ₆ cycloalkyl", and the term "C ₃ -C ₆ cycloalkyl" is understood to mean a saturated monocyclic or bicyclic hydrocarbon ring having 3 to 6 carbon atoms, specific examples include but are not limited to cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, etc.

The term "cycloalkenyl" refers to a non-aromatic carbon ring that is not fully saturated and exists in the form of a single ring, a branched ring, a bridged ring or a spiro ring. Unless otherwise indicated, the carbocyclic ring is generally 3 to 10 membered. "C ₃ -C ₁₀ cycloalkenyl" may include "C ₄ -C ₁₀ cycloalkenyl" and "C ₅ -C ₈ cycloalkenyl". Specific examples of the cycloalkenyl group include, but are not limited to, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, cycloheptenyl or cycloheptadienyl, and the like.

The term "heterocyclyl" refers to a fully saturated or partially saturated (not aromatic heteroaromatic as a whole) monocyclic, paracyclic, spirocyclic or bridged cyclic group containing 1 to 5 ring atoms. Heteroatom or heteroatom group (that is, an atomic group containing heteroatoms), the "heteroatom or heteroatom group" includes but is not limited to nitrogen atom (N), oxygen atom (O), sulfur atom (S), phosphorus atom (P) , boron atom (B), -S(=O) ₂ -, -S(=O)-, -P(=O) ₂ -, -P(=O)-, -NH-, -S(=O )(=NH)-, -C(=O)NH- or -NHC(=O)NH-, etc. The term "3-10 membered heterocyclyl" refers to a heterocyclyl with a number of ring atoms of 3, 4, 5, 6, 7, 8, 9 or 10, and its ring atoms contain 1 to 5 independently selected from the above The heteroatom or heteroatom group. "3-10-membered heterocyclyl" includes "4-7-membered heterocyclyl", wherein specific examples of 4-membered heterocyclyl include but are not limited to azetidinyl or oxetanyl; 5-membered heterocyclyl Heterocyclyl Specific examples include, but are not limited to, tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl, 4,5-dihydroxazolyl or 2,5-dihydroxazolyl. Hydrogen-1H-pyrrolyl; specific examples of 6-membered heterocyclic groups include, but are not limited to, tetrahydropyranyl, piperidyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, trithianiyl Alkyl, tetrahydropyridyl or 4H-[1,3,4]thiadiazinyl; specific examples of 7-membered heterocyclyl include but are not limited to diazepanyl. The heterocyclic group may also be a bicyclic group, wherein specific examples of 5,5-membered bicyclic groups include but are not limited to hexahydrocyclopenta[c]pyrrole-2(1H)-yl; 5,6-membered bicyclic groups. Specific examples include, but are not limited to, hexahydropyrro[1,2-a]pyrazin-2(1H)-yl, 5,6,7,8-tetrahydro-[1,2,4]triazolo[4 ,3-a]pyrazinyl or 5,6,7,8-tetrahydroimidazo[1,5-a]pyrazinyl. Optionally, the heterocyclic group may be a benzo-fused cyclic group of the above-mentioned 4-7 membered heterocyclic group, and specific examples include but are not limited to dihydroisoquinolyl and the like. "4-10-membered heterocyclyl" may include "5-10-membered heterocyclyl", "4-7-membered heterocyclyl", "5-6-membered heterocyclyl", "6-8-membered heterocyclyl" , "4-10 membered heterocycloalkyl", "5-10 membered heterocycloalkyl", "4-7 membered heterocycloalkyl", "5-6 membered heterocycloalkyl", "6-8 membered "Heterocycloalkyl" and other scopes, "4-7 membered heterocyclyl" may further include "4-6 membered heterocyclyl", "5-6 membered heterocyclyl", "4-7 membered heterocyclyl" , "4-6 membered heterocycloalkyl", "5-6 membered heterocycloalkyl" and other ranges. Although some bicyclic heterocyclyl groups in the present disclosure partially contain a benzene ring or a heteroaromatic ring, the heterocyclyl groups are still non-aromatic as a whole.

The term "aryl" refers to an all-carbon monocyclic or fused polycyclic aromatic ring group having a conjugated π electron system. Aryl groups can have 6-20 carbon atoms, 6-14 carbon atoms, or 6-12 carbon atoms. The term "C ₆ -C ₂₀ aryl" is understood to mean an aryl group having 6 to 20 carbon atoms. In particular, a ring with 6 carbon atoms ("C ₆ aryl"), for example phenyl; or a ring with 9 carbon atoms ("C ₉ aryl"), for example indanyl or indenyl; or a ring with 10 or a ring of 13 carbon atoms ("C ₁₀ aryl"), such as tetrahydronaphthyl, dihydronaphthyl or naphthyl; or a ring of 13 carbon atoms ("C ₁₃ aryl"), such as fluorenyl; or is a ring having 14 carbon atoms ("C ₁₄ aryl"), such as anthracenyl. The term "C ₆ -C ₁₀ aryl" is understood to mean an aryl group having 6 to 10 carbon atoms. In particular, a ring with 6 carbon atoms ("C ₆ aryl"), for example phenyl; or a ring with 9 carbon atoms ("C ₉ aryl"), for example indanyl or indenyl; or a ring with 10 A ring of 10 carbon atoms ("C ₁₀ aryl"), such as tetrahydronaphthyl, dihydronaphthyl or naphthyl. The term "C ₆ -C ₂₀ aryl" may include "C ₆ -C ₁₀ aryl"

The term "heteroaryl" refers to an aromatic monocyclic or fused polycyclic ring system containing at least one ring atom selected from N, O, and S, and the remaining ring atoms are C. The term "5-10 membered heteroaryl" is understood to include monocyclic or bicyclic aromatic ring systems having 5, 6, 7, 8, 9 or 10 ring atoms, in particular 5 or 6 or 9 or 10 ring atoms, and it contains 1-5, preferably 1-3 heteroatoms independently selected from N, O and S. In particular, the heteroaryl group is selected from the group consisting of thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl or thiazolyl Diazolyl, etc. and their benzo derivatives, such as benzofuryl, benzothienyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzotriazole base, indazolyl, indolyl or isoindolyl, etc.; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl or triazinyl, etc. and their benzo derivatives, such as quinolyl, quinazole Phylline or isoquinolyl, etc.; or azocinyl, indolizinyl, purinyl, etc. and their benzo derivatives; or cinnolinyl, phthalazinyl, quinazoline base, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl or phenoxazinyl, etc. The term "5-6 membered heteroaryl" refers to an aromatic ring system having 5 or 6 ring atoms and containing 1-3, preferably 1-2 heteroatoms independently selected from N, O and S.

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

The term "therapeutically effective amount" means

(i) treat a particular disease, condition, or disorder, (ii) reduce, ameliorate, or eliminate one or more symptoms of a particular disease, condition, or disorder, or (iii) delay the onset of a particular disease, condition, or disorder described herein The amount of a compound of the present disclosure that is responsible for the onset of one or more symptoms.

The amount of a compound of the present disclosure that constitutes a "therapeutically effective amount" will vary depending on the compound, the disease state and its severity, the mode of administration, and the age of the mammal to be treated, but can be routinely determined by one skilled in the art. based on its own knowledge and the contents of this disclosure.

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 multiple toxicity, irritation, allergic reactions, or other problems or complications, commensurate with a reasonable benefit/risk ratio.

The term "pharmaceutically acceptable salts" refers to salts of pharmaceutically acceptable acids or bases, including salts of compounds with inorganic or organic acids, and salts of compounds with inorganic or organic bases.

The term "pharmaceutical composition" refers to a mixture of one or more compounds of the present disclosure or salts thereof and pharmaceutically acceptable excipients. The purpose of pharmaceutical compositions is to facilitate administration of the compounds of the present disclosure to an organism.

The term "pharmaceutically acceptable excipients" refers to those excipients that have no obvious irritating effect on the organism and do not impair the biological activity and performance of the active compound. Suitable excipients are well known to those skilled in the art, such as carbohydrates, waxes, water-soluble and/or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, etc.

The words "comprise" or "comprise" and their English variations such as compris or comprising can be understood as having an open, non-exclusive meaning, that is, "including but not limited to".

The present disclosure also includes isotopically labeled compounds that are the same as those described herein, but in which one or more atoms are replaced by an atom having an atomic weight or mass number different from that typically found in nature. Examples of isotopes that may be incorporated into the compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as ² H, ³ H, ¹¹ C, ¹³ C, ¹⁴ C, ¹³ respectively N, ¹⁵ N, ¹⁵ O, ¹⁷ O, ¹⁸ O, ³¹ P, ³² P, ³⁵ S, ¹⁸ F, ¹²³ I, ¹²⁵ I and ³⁶ Cl, etc.

Certain isotopically labeled compounds of the present disclosure (eg, labeled with ³ H and ¹⁴ C) can be used in compound and/or substrate tissue distribution analyses. Tritiated (ie ³ H) and carbon-14 (ie ¹⁴ C) isotopes are particularly preferred due to their ease of preparation and detectability. Positron-emitting isotopes such as ¹⁵ O, ¹³ N, ¹¹ C, and ¹⁸ F can be used in positron emission tomography (PET) studies to determine substrate occupancy. This can generally be accomplished by the following procedures similar to those disclosed in the Schemes and/or Examples below, Isotopically labeled compounds of the present disclosure are prepared by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

The pharmaceutical compositions of the present disclosure can be prepared by combining the compounds of the present disclosure with suitable pharmaceutically acceptable excipients, for example, they can be formulated into solid, semi-solid, liquid or gaseous preparations, such as tablets, pills, capsules, powders , granules, ointments, emulsions, suspensions, suppositories, injections, inhalants, gels, microspheres and aerosols, etc.

Typical routes of administration of the compounds of the present disclosure, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, Intramuscular, subcutaneous, and intravenous administration.

The pharmaceutical composition of the present disclosure can be manufactured using methods well known in the art, such as conventional mixing methods, dissolution methods, granulation methods, emulsification methods, freeze-drying methods, etc.

In some embodiments, the pharmaceutical composition is in an oral form. For oral administration, the pharmaceutical compositions may be formulated by mixing the active compounds with pharmaceutically acceptable excipients well known in the art. These excipients enable the compounds of the present disclosure to be formulated into tablets, pills, dragees, dragees, capsules, liquids, gels, slurries, suspensions, etc., for oral administration to patients.

Solid oral compositions may be prepared by conventional mixing, filling or tableting methods. For example, it can be obtained by the following method: mixing the active compound with solid excipients, optionally grinding the resulting mixture, adding other suitable excipients if necessary, and then processing the mixture into granules to obtain tablets Or sugar-coated core. Suitable excipients include, but are not limited to: binders, diluents, disintegrants, lubricants, glidants or flavoring agents, etc.

The pharmaceutical compositions may also be suitable for parenteral administration as sterile solutions, suspensions or lyophilized products in suitable unit dosage forms.

In all methods of administration of the compounds of general formula (I) described herein, dosages of 0.01 mg/kg to 200 mg/kg body weight are administered daily, in single or divided doses.

Detailed ways

The present disclosure will be described in detail below with reference to examples, but the following examples should not be regarded as limiting the scope of the present disclosure.

Example 1 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-6-methyl-8-(4-(1-methyl-4-(trifluoromethyl)-1H -Imidazol-2-yl)benzyl)pyrido[4,3-d]pyrimidin-7(6H)-one (Compound 1)

Step 1: 2,4-dichloro-5-(dimethoxymethyl)pyrimidine (1-2)

Compound 2,4-dichloropyrimidine-5-carbaldehyde (1.75g, 10mmol, 1eq) and trimethyl orthoformate (2.12g, 20mmol, 2eq) were dissolved in methanol (20mL) and stirred at 65°C for 5 hours. . The solvent was evaporated from the reaction solution under reduced pressure, and water (50 mL) was added to the residue. The resulting mixture was extracted with ethyl acetate (50 mL × 3 times). The organic phases were mixed, washed with saturated brine (30 mL × 3 times), dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The obtained residue was purified using a silica gel column (the eluent phase was a mixed solvent of petroleum ether containing 30% ethyl acetate) to obtain the title compound 1-2 (1.82g, 8.2mmol, yield: 82%) as a white oil.

Step 2: Diethyl 2-(2-chloro-5-(dimethoxymethyl)pyrimidin-4-yl)malonate (1-3)

Compound 2B (1.57g, 9.84mmol, 1.2eq) was dissolved in tetrahydrofuran (20mL), sodium hydride (0.59g, 9.84mmol, 1.2eq) was added at 0°C, and stirred for 0.5 hours. Then add the reactant 2,4-dichloro-5-(dimethoxymethyl)pyrimidine (1.82g, 8.2mmol, 1eq), heat the reaction to 80°C for 2 hours, and then cool the reaction solution to room temperature. The solvent was distilled off under reduced pressure, and water (30 mL) was added to the residue. The resulting mixture was extracted with ethyl acetate (30 mL × 3 times). The organic phases were mixed, washed with saturated brine (30 mL × 3 times), dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The obtained residue was purified using a silica gel column (the eluent phase was a mixed solvent of petroleum ether containing 20% ethyl acetate) to obtain the title compound 1-3 (1.9g, 5.5mmol, yield: 67%) as a white oil.

Step 3: Ethyl 2-(2-chloro-5-(dimethoxymethyl)pyrimidin-4-yl)acetate (1-4)

Compound 1-3 (1.9g, 5.5mmol, 1.0eq) was dissolved in ethanol (30mL), sodium ethoxide (0.025g, 0.55mmol, 0.1eq) was added, heated to 85°C and stirred for 10 hours. The reaction solution was then cooled to room temperature, the solvent was evaporated under reduced pressure, and water (50 mL) was added to the residue. The resulting mixture was extracted with ethyl acetate (30 mL × 3 times). The organic phases were mixed, washed with saturated brine (30 mL × 3 times), dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The obtained residue was purified using a silica gel column (the eluent phase was a mixed solvent of petroleum ether containing 35% ethyl acetate) to obtain the title compound 1-4 (0.61g, 2.25mmol, yield: 41%) as a white oil.

Step 4: 2-(4'-cyclopropyl-5-(dimethoxymethyl)-6'-methoxy-[2,5'-bipyrimidin]-4-yl)acetate (1 -5)

Compound 1-4 (0.61g, 2.25mmol, 1.0eq) and compound 4B (0.57g, 2.92mmol, 1.3eq) were dissolved in 1,4-dioxane (5mL) and water (0.5mL), and added Chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl) ] Palladium (II) (0.2g, 0.25mmol, 0.1eq), potassium phosphate (0.95g, 4.5mmol, 2eq), then the reaction solution was heated to 100°C and stirred for 3 hours. The reaction solution was then cooled to room temperature, the solvent was evaporated under reduced pressure, and water (50 mL) was added to the residue. The resulting mixture was extracted with ethyl acetate (30 mL × 3 times). The organic phases were mixed, washed with saturated brine (30 mL × 3 times), dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The obtained residue was purified using a silica gel column (the eluent phase was a mixed solvent of petroleum ether containing 80% ethyl acetate) to obtain the title compound 1-5 as a white solid (0.63g, 1.62mmol, yield: 72%).

Step 5: 2-(4'-cyclopropyl-5-(dimethoxymethyl)-6'-methoxy-[2,5'-bipyrimidin]-4-yl)-3-(4 -(1-Methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)propionic acid ethyl ester (1-6)

Dissolve compound 1-5 (0.39g, 1mmol, 1.0eq) in N,N-dimethylformamide (10mL), add sodium hydride (0.05g, 1.2mmol, 1.2eq) at 0°C, and stir 0.5 hours. Then reactant 5B (0.38g, 1.2mmol, 1.2eq) was added, reacted for 2 hours, the solvent was distilled off under reduced pressure, and water (30mL) was added to the residue. The resulting mixture was extracted with ethyl acetate (30 mL × 3 times). The organic phases were mixed, washed with saturated brine (30 mL × 3 times), dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The obtained residue was purified using a silica gel column (the eluent phase was a mixed solvent of petroleum ether containing 60% ethyl acetate) to obtain the title compound 1-6 (0.32g, 0.51mmol, yield: 51%) as a light yellow oil.

Step 6: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-6-methyl-8-(4-(1-methyl-4-(trifluoromethyl)-1H -Imidazol-2-yl)benzyl)pyrido[4,3-d]pyrimidin-7(6H)-one (Compound 1)

Dissolve compound 1-6 (0.12g, 0.2mmol, 1.0eq) in toluene (2mL) and acetic acid (2mL), then add the reactant methylamine hydrochloride (0.02g, 0.6mmol, 3eq), and heat the reaction solution to The reaction was carried out at 110°C for 2 hours, the solvent was evaporated under reduced pressure, and water (30 mL) was added to the residue. The resulting mixture was extracted with ethyl acetate (30 mL × 3 times). The organic phases were mixed, washed with saturated brine (30 mL × 3 times), dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The obtained residue was purified by preparative chromatography (Waters Xbridge C18, mobile phase 10-90% acetonitrile aqueous solution gradient elution) to obtain the title compound 1 (21 mg, yield 19%) as a yellow solid. m/z(ESI):548.2[M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ) δ9.29 (s, 1H), 9.24 (s, 1H), 8.70 (s, 1H), 7.88 (s, 1H), 7.53 (d, J = 8.0Hz, 2H),7.42(d,J=8.1Hz,2H),4.20(s,2H),3.90(s,3H),3.78(s,3H),3.72(s,3H),1.86(s,1H), 1.07(s,2H),0.89(s,2H).

Example 2

Using similar synthesis steps and methods in Example 1, and using different raw materials a in the table below to replace methylamine hydrochloride in step 6 in Example 1, compounds 2 to 12 in the table below can be synthesized.

Example 3

Compounds 13 to 17 in the following table can be synthesized by using similar synthetic steps and methods in Example 1 and using different raw materials b in the following table to replace compound 5B in step 5 in Example 1.

Example 4

Compound 18 in the following table can be synthesized by using similar synthetic steps and methods in Example 1 and using different raw materials c in the following table instead of compound 4B in step 4 in Example 1.

Example 5 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-6-difluoromethyl-8-(4-(1-methyl-4-(trifluoromethyl)) -1H-imidazol-2-yl)benzyl)pyrido[4,3-d]pyrimidin-7(6H)-one (compound 19)

Compound 2 (20.0 mg, 0.037 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide (1 mL), potassium carbonate (15.5 mg, 0.11 mmol, 3 eq) was added, and then compound 19a (17.1 mg, 0.11mmol, 3eq), heat the reaction solution to 80°C for 2 hours. The obtained reaction solution was directly purified by preparative chromatography (Waters Xbridge C18, mobile phase 10-90% acetonitrile aqueous solution gradient elution) to obtain the solid title compound 19 (7 mg, yield 32%). m/z(ESI):584.2[M+H] ⁺ .

¹ H NMR(400MHz, DMSO-d ₆ )δ9.96(s,1H),9.43(s,1H),8.75(s,1H),7.89(s,1H),7.96-7.94(m,1H), 7.60-7.56(m,2H),7.41-7.38(m,2H),4.51-4.46(m,2H),3.88(s,3H),3.72(s,3H),1.80-1.74(m,1H), 1.09-1.06(m,2H),0.89-0.82(m,2H).

Test Example 1: USP1 enzyme in vitro activity detection experiment

laboratory apparatus:

Experimental Materials:

The USP1 enzyme (Recombinant Human His6-USP1/His6-UAF1 Complex Protein, CF) used in the experiment was purchased from R&D Systems, catalog number E-568-050. After aliquot, store at -80℃.

The detection kit (Ub-CHOP2-Reporter Deubiquitination Assay Kit) was purchased from Lifesensors Company, product number is PR1101. After aliquot, store at -80℃. The kit contains a ubiquitinated reporter enzyme. When deubiquitinated by USP1/UAF1, it becomes active. After catalyzing the substrate, the substrate is excited by a 485nm laser to produce a 535nm emission light signal.

Information on other reagents and consumables required for the experiment is as follows:

experimental method:

Compounds to be tested were dissolved in DMSO to 10mM. Use the ECHO instrument to pour the compound and pure DMSO into each well of the 384-well plate. The highest concentration starts from 3 μM and is diluted 3 times for a total of 8 concentration points. Add 50nL of the compound to be tested or DMSO (as a control) to each well. The instrument Gradient dilutions of sample concentrations were obtained through different ratios (3000nM, 1000nM, 333nM, 111nM, 37nM, 12nM, 4.1nM and 1.4nM). Dilute the enzyme with freshly prepared reaction solution (20mM Tris-HCl (pH 8.0), 2mM CaCl ₂ , 2mM β-mercaptoethanol, 0.05% CHAPS (dilute CHAPS with ddH ₂ O)). Add 5 μL of diluted enzyme reaction solution to each well, centrifuge and shake to mix the enzyme and compound, centrifuge again and place on ice. Dilute the kit reporter system and substrate with the reaction solution, add 5 μL of diluted liquid to each well, and centrifuge to mix. Incubate at room temperature for 0.5 hours. The fluorescence signal in each well was measured using a plate reader (excitation light wavelength 485 nm, emission light wavelength 535 nm). The IC ₅₀ value of the compound's inhibitory activity on enzyme activity was calculated using the four-parameter Logistic Model method. In the following formula, x represents the logarithmic form of the compound concentration; F(x) represents the effect value (inhibition rate of enzyme activity under this concentration): F(x)=(A+((BA)/(1+((C /x)^D)))). A, B, C and D are four parameters. IC ₅₀ values were further calculated into best fit curves using Xlfit Concentration of compound required for 50% inhibition of enzyme activity.

The test results are shown in Table 1.

Table 1 USP1 enzyme inhibitory activity in vitro

Test Example 2: Inhibition experiment of the disclosed compounds on MDA-MB-436 cell proliferation

Detection based on CellTiter-Glo luminescent live cell detection system.

laboratory apparatus:

Experimental Materials:

The cells used in the experiment, MDA-MB-436, were purchased from Kebai Biotechnology Co., Ltd., product number CBP60385. The cells were subcultured in culture medium (DMEM containing 10% FBS) and frozen in liquid nitrogen when the cell passage number was low. The cells used in the experiment did not exceed 15 generations.

Detection kit ( Luminescent Cell Viability Assay) was purchased from Promega, Cat. No. for G7573. After aliquot, store at -30℃. The kit is a homogeneous detection method for detecting the number of viable cells in culture by quantitatively measuring ATP. The kit produces a luminescent signal proportional to the amount of ATP present, which is directly proportional to the number of cells in the culture.

Information on other reagents and consumables required for the experiment is as follows:

experimental method:

Digest the cultured cells with 0.25% Trypsin-EDTA Solution, collect and centrifuge, adjust the concentration and resuspend in culture medium (DMEM containing 10% FBS), seed the cells in a 384-well plate (400 cells/20 μL/well), 37°C , culture in a 5% _CO2 cell culture incubator overnight. Use the Echo sonic pipetting system to add the compound to be tested into the 384-well plate. The highest concentration starts from 10 μM and is diluted 4 times for a total of 8 concentration points. Add 100 nL of the compound to be tested or DMSO (as a control) to each well to obtain a gradient dilution. Sample concentrations (10000nM, 2500nM, 625nM, 156nM, 39nM, 10nM, 2.4nM and 0.61nM). Add 30 μL of culture medium to each well, mix by centrifugation, and then centrifuge again, then culture in a cell culture incubator for 7 days (one row of cells will be added with CTG (CellTiter-Glo) detection solution on the day of drug addition). After the 7th day, add 25 μL CTG detection solution to each well, mix by centrifugation, and then centrifuge and place at room temperature in the dark for 10 minutes. The chemiluminescence signal was measured in each well using a plate reader (emission wavelength 400-700nm). The chemical luminescence value [RLU] cpd was obtained for 7 days in the drug-added group, and the chemical luminescence value [RLU]cpd was obtained in the DMSO-only group without drug addition for 7 days. Luminescence value [RLU] cell, parallel CTG test on day 0 of the DMSO-free group obtained the chemiluminescence value [RLU] background on day 0. Inhibition rate of compound on proliferation Inhibition rate (%) = [1-([RLU]cpd–[RLU]background)/([RLU]cell–[RLU]background)]×100%, inhibitory activity of compound on proliferation GI _{The 50} value is calculated using the four-parameter Logistic Model method. In the following formula, x represents the logarithmic form of the compound concentration; F(x) represents the effect value (inhibition rate of proliferation under this concentration): F(x)=(A+((BA)/(1+((C/ x)^D)))). A, B, C and D are four parameters. The GI ₅₀ value was further calculated as the compound concentration required for 50% inhibition of proliferation in the best-fit curve using Xlfit.

The inhibitory activity of the compounds of the present disclosure on the proliferation of MDA-MB-436 was measured through the above test. The measured GI ₅₀ value is shown in Table 2.

Table 2 The inhibitory activity of the disclosed compounds on MDA-MB-436 cell proliferation

Test Example 3: CYP enzyme inhibition test of compounds of the present disclosure

150 donor pooled human liver microsomes (purchased from Corning, Cat. No. 452117) were used to evaluate representative substrate metabolic responses of the five major human CYP isoforms (CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP3A4/5). Liquid chromatography tandem mass spectrometry (LC/MS/MS) was used to determine the response of test compounds at different concentrations to phenacetin (CYP1A2), diclofenac sodium (CYP2C9), S-mephenytoin (CYP2C19), and bufuralol hydrochloride. Effects of salt (CYP2D6) and midazolam (CYP3A4/5) metabolic reactions.

30μM phenacetin, 10μM diclofenac sodium, 35μM S-mephenytoin, 5μM bufurolol hydrochloride, 3μM midazolam, 1mM reduced nicotinamide adenine dinucleotide phosphate (NADPH), The concentration of the compound to be tested is 0.1, 0.3, 1, 3, 10, 30 μmol/L or the reaction system of positive compound or blank control and mixed human liver microsomes (0.2mg/mL) is 200μL (100mmol/L phosphate buffer, pH 7.4, containing 0.3% DMSO, 0.6% acetonitrile, and 0.1% methanol by volume) and incubated at 37°C for 5 minutes. Then add 200 μL of acetonitrile solution containing 3% formic acid and 40 nM internal standard verapamil, and centrifuge at 4000 rpm for 50 minutes. Cool on ice for 20 minutes, and centrifuge at 4000 rpm for 20 minutes to precipitate protein. Take 200 μL of supernatant for LC-MS/MS analysis.

Peak areas were calculated from the chromatograms.

The residual activity ratio (%) is calculated using the following formula:

Peak area ratio = metabolite peak area/internal standard peak area

Residual activity ratio (%) = peak area ratio of the compound group to be tested/peak area ratio of the blank group

The CYP half inhibitory concentration (IC ₅₀ ) was calculated by Excel XLfit 5.3.1.3.

The measured CYP half-inhibitory concentration (IC ₅₀ ) values of the disclosed compounds are shown in Table 3.

Table 3. Half-time inhibitory concentration (IC ₅₀ ) of CYP of compounds of the present disclosure

Test Example 4: Caco-2 permeability test of compounds of the present disclosure

The apparent permeability coefficient (P _app ) of the drug was measured and analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS) in Caco-2 cell model.

In this test example, Caco-2 cells were purchased from the American Type Culture Collection (ATCC), 4-hydroxyethylpiperazineethanesulfonic acid (HEPES) was purchased from Beijing Solebao Technology Co., Ltd., and Hank's Balanced Salt Solution ( HBSS) and non-essential amino acids (NEAA) were purchased from Thermo Fisher Scientific, penicillin, streptomycin and trypsin/EDTA were purchased from Solebac, fetal bovine serum (FBS) and DMEM culture medium were purchased from Corning Company , HTS-96-well Transwell plate and other sterile consumables were purchased from Corning Company, Millicell resistance measurement system was purchased from Millipore, K2 was purchased from Nexcelom Bioscience, the Infinite 200 PRO microplate reader was purchased from Tecan, and the MTS2/4 orbital shaker was purchased from IKA Labortechnik.

The first step: cell culture and seeding plate

Caco-2 was cultured in a cell culture flask, and the incubator was set to 37°C, 5% CO ₂ , and a relative humidity of 95%. Cells can be used to seed Transwell plates when they reach 70-90% confluence. Before cell seeding, add 50 μL of cell culture medium to each well of the Transwell plate, and add 25 mL of cell culture medium to the lower culture plate. Place the culture plate in a 37°C, 5% _CO2 incubator and incubate it for 1 hour before seeding cells. After cell digestion, transfer the cell suspension to a round-bottomed centrifuge tube and centrifuge at 120g for 5 minutes. Resuspend cells in cell culture medium to a final concentration of 6.86×10 ⁵ cells/mL. The cell suspension was added to the 96-well Transwell plate chamber at 50 μL/well, and the final seeding density was 2.4×10 ⁵ cells/cm ² . Start changing the medium 24 hours after inoculation, culture for 14-18 days, and change the medium every other day.

Step 2: Evaluation of cell monolayer membrane integrity

Caco-2 reaches confluence and completes differentiation after approximately 14 days of culture. At this time, it can be applied to penetration testing. The resistance of the single layer membrane was measured with a resistometer (Millipore, USA), and the resistance of each hole was recorded. After the measurement is completed, it is returned to the incubator. Calculation of resistance value: Measure resistance value (ohms) × membrane area (cm ² ) = TEER value (ohm·cm ² ). If TEER value is <230ohms·cm ² , the hole cannot be used for penetration test.

Step 3 Solution Preparation

Weigh 2.38g HEPES and 0.35g sodium bicarbonate respectively, add 900mL pure water to dissolve, then add 100mL 10×HBSS, stir evenly, adjust the pH to 7.4, and finally filter to obtain 1L transport buffer (HBSS, 10mM HEPES, pH 7.4 ).

Dilute the 1 mM DMSO solution stock solution of the compound to be tested with transport buffer to obtain a 5 μM test solution. The control compound digoxin was diluted to 2mM with DMSO and diluted to 10μM with the above transport buffer to obtain a control compound test solution. In addition, DMSO was also diluted with the above transport buffer to a receiving end solution containing 0.5% DMSO.

Step 4 Drug Penetration Test

Remove the Transwell plate from the incubator. Use transport buffer solution (10mM HEPES, pH 7.4) to rinse the cell monolayer twice and incubate at 37°C for 30 minutes.

Determine the transport rate of compounds from the apical to the basal end. Add 125 μL test solution to each well of the upper chamber (top). And immediately transfer 50 μL of the solution from the tip to 200 μL of acetonitrile containing the internal standard (0.1 μM tolbutamide) as the initial sample from the tip to the base. Add 235 μL of receiving end solution to each well in the lower chamber (basal end).

Determine the rate of compound transport from basal to apical end. Add 285 μL of the receiving end solution to each well of the upper chamber (top), and immediately transfer 50 μL of the solution from the top to 200 μL of acetonitrile containing the internal standard (0.1 μM tolbutamide) as the initial sample from the base to the top. Add 75 μL test solution to each well in the lower chamber (basal end).

After combining the upper and lower transport devices, incubate at 37°C for 2 hours.

After the incubation is completed, take 50 μL of each well from the upper and lower chambers of the Transwell plate and add it to a new sample tube. Add 200 μL of acetonitrile containing internal standard (0.1 μM tolbutamide) into the sample tube, vortex for 10 minutes, and centrifuge at 3220 g for 40 minutes. Aspirate 150 μL of the supernatant, dilute it with 150 μL of water, and perform LC-MS/MS analysis. All samples were prepared in triplicate.

The integrity of the cell monolayer after 2 hours of incubation was evaluated by the leakage of Lucifer Yellow, and the Lucifer Yellow stock solution was diluted using transport buffer solution (10mM HEPES, pH 7.4) to a final concentration of 100 μM. Add 100 μL of Lucifer Yellow solution to each well of the upper insert plate of the Transwell, and add 300 μL of transport buffer solution (10mM HEPES, pH 7.4) to each well of the lower receiving plate. After incubating at 37°C for 30 minutes, pipet 80 μL of the solution from the upper and lower layers of each well into a new 96-well plate. Fluorescence measurement was performed using a microplate reader with an excitation wavelength of 485 nm and an emission wavelength of 530 nm.

Step 5 Data Analysis

All calculations were performed using Microsoft Excel. Peak areas were determined using extracted ion chromatograms.

The apparent permeability coefficient (P _app , unit: cm/s×10 ^-6 ) is calculated using the following formula:

In the formula: V _A is the volume of the receiving end solution (Ap→Bl is 0.3mL, Bl→Ap is 0.1mL), Area is the Transwell-96-well plate membrane area (0.143cm ² ); time is the incubation time (unit: s ); [drug] _receiver is the drug concentration at the receiving end; [drug] _{initial, donor} is the initial drug concentration at the giving end. .

The efflux rate is calculated using the following formula:

In the formula: P _{app (BA)} is the apparent permeability coefficient from the base end to the top;

P _app(AB) is the apparent permeability coefficient from the tip to the base.

The recovery rate is calculated using the following formula:

In the formula: V _A is the solution volume at the receiving end (unit: mL); V _D is the solution volume at the giving end (unit: mL).

Leakage rate is calculated using the following formula:

In the formula: I _receiver refers to the fluorescence density of the receiving hole (0.3mL), I _donor refers to the fluorescence density of the dosing hole (0.1mL), expressed in LY (%). LY<1.5% indicates that the single-layer cell membrane is intact.

The Caco-2 permeability data obtained by testing the disclosed compounds are shown in Table 4.

Table 4. Caco-2 permeability data for compounds of the present disclosure

Test Example 5: In vitro metabolic stability detection of rat liver cells of the disclosed compound

Use LC/MS/MS to measure the concentration of the compound in the reaction system to calculate the intrinsic clearance rate of the compound to be tested and evaluate its in vitro metabolic stability in rat hepatocytes.

Add 198 μL of 0.5 × 10 ⁶ cells/mL rat hepatocyte mixture and 2.0 μL of 100 μM test compound or positive control (verapamil) to the incubation plate to start the reaction. Incubation was performed at 37°C and 900 rpm. Transfer 25 μL of the incubation system to the stop plate (150 μL of acetonitrile containing 100 nM alprazolam, 200 nM caffeine, and 100 nM tosylbutamide per well) at 0, 15, 30, 60, 90, and 120 minutes. Then vortex for 5 minutes. Centrifuge the stop plate at 3220g for 45 minutes. Transfer 100 μL of the supernatant of each compound to a 96-well injection plate, and then add 100 μL of pure water to dilute the sample.

The resulting samples were quantified from ion chromatograms. Calculate the residual rate based on the peak area of the test compound or positive control. The slope k was determined from a linear regression of the natural logarithm of the residual rate versus incubation time using Microsoft Excel.

Intrinsic clearance (in vitro CL _int , μL/min/ ¹⁰ cells) was calculated from the slope value according to the following equation:

in vitro CL _int =-kV/N

V=incubation volume (0.25mL);

N = number of cells per well (0.125×10 ⁶ cells)

The measured intrinsic clearance rate values of rat hepatocytes are shown in Table 5.

Table 5. Intrinsic clearance rate of rat hepatocytes of the disclosed compounds

Test Example 6: Compound solubility (PBS pH 7.4) test

The solubility of the test compound in PBS pH 7.4 was determined using LC/MS/MS.

Take 6μL of 10mM DMSO solution of the compound to be tested and mix it with 194μL DMSO to prepare a 300μM compound solution. Take 5 μL of this solution and mix it with 5 μL of PBS pH 7.4 solution in 490 μL of water and acetonitrile (1:1) containing the internal standard to prepare a 3 μM standard concentration solution of the sample to be tested. The dilution factor of the standard solution can be adjusted according to the LC/MS response signal strength.

Take 15 μL of 10 mM DMSO solution of the compound to be tested and add it to 485 μL of PBS pH 7.4 solution. Add a stir bar and shake the solubility sample bottle at 25°C and 1100 rpm for 2 hours. After shaking, remove the stirrer, transfer the sample to a filter plate and filter using a vacuum manifold. Take 5 μL of the filtrate and mix it with 5 μL of PBS pH 7.4 solution in 490 μL of water and acetonitrile (1:1) containing the internal standard to prepare the test solution. The dilution factor can be adjusted based on the solubility value and LC/MS response signal strength.

The obtained samples were detected by LC/MS/MS. Calculate the sample solubility based on the peak areas of the test compound solution and the standard concentration solution. Calculated as follows:

[Sample] is the solubility of the sample to be tested;

Area ratio _sample is the ratio of the sample peak area to the internal standard peak area in the sample to be tested;

INJ VOL STD is the injection volume of standard concentration solution;

DF _sample is the dilution factor of the sample solution to be tested;

[STD] is the concentration of standard concentration solution;

INJ VOL _sample is the injection volume of the sample solution to be tested;

Area ratio STD is the ratio of the sample peak area to the internal standard peak area in the standard concentration solution.

The solubility of the compounds measured by this method is shown in Table 6.

Table 6. Solubility of compounds of the present disclosure in PBS pH 7.4

Claims (18)

1. a compound of formula (I) or a pharmaceutically acceptable salt thereof,
   

   in, Represents a single or double bond;

   X is selected from N, CR ³ , NR ³ or C(O);

   Y is selected from NR ⁴ or C(O);

   Z is selected from CR ⁵ or NR ⁵ ;

   R ³ , R ⁴ , R ⁵ are independently selected from H, halogen, CN, OH, NH ₂ , -C(O)OR ^x , -C(O)R ^x , -OC ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₁₀ cycloalkyl or 4-7 membered heterocyclyl, the NH ₂ , -OC ₁ -C ₆ Alkyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₁₀ cycloalkyl or 4-7 membered heterocyclyl is optionally substituted by R ^x ;

   Ring A is selected from C _{6 -C 10} aryl or _5-10 membered heteroaryl, which is optionally substituted by one or more R ^a ;

   Ring B is selected from C ₆ -C ₁₀ aryl, 5-10 membered heteroaryl, 4-10 membered heterocyclyl, C ₃ -C ₁₀ cycloalkenyl or C ₃ -C ₁₀ cycloalkyl, the C ₆ -C ₁₀ aryl, 5-10 membered heteroaryl, 4-10 membered heterocyclyl, C ₃ -C ₁₀ cycloalkenyl or C ₃ -C ₁₀ cycloalkyl optionally substituted by one or more R ^b ;

   Ring C is selected from C ₆ -C ₁₀ aryl _{or 5-10} membered heteroaryl, which is optionally substituted by one or more R ^c ;

   Each R ^a , R ^b , R ^c is independently selected from halogen, CN, OH, NH ₂ , -C(O)OR ^x , C ₁ -C ₆ alkyl, -OC ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl or 4-10 membered heterocyclyl, the NH ₂ , C ₁ -C ₆ alkyl, -OC ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl or 4-10 membered Heterocyclyl is optionally substituted with ^Rx ,

   Or R ^b , R ^c and the atoms to which they are connected together form a C ₃ -C ₁₀ cycloalkenyl group or a 4-10 membered heterocyclyl group, and the C ₃ -C ₁₀ cycloalkenyl group or 4-10 membered heterocyclyl group can be any The chosen land is replaced by R ^x ;

   R ¹ and R ² are independently selected from H, halogen, CN, OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl or 4-10 membered heterocyclyl, the OH, NH _2. C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl or 4-10 membered heterocyclyl is optionally substituted by R ^x ,

   Or R ¹ , R ² and the atoms to which they are connected together form a C ₃ -C ₁₀ cycloalkyl group or a 4-7 membered heterocyclyl group, and the C ₃ -C ₁₀ Cycloalkyl or 4-7 membered heterocyclyl is optionally substituted by ^Rx ;

   ^R _{_ _ _ _ _ _ _ _ _} C ₁₀ cycloalkyl or 4-7 membered heterocyclyl substituted.
2. The compound represented by formula (I) or a pharmaceutically acceptable salt according to claim 1, characterized in that: R ³ , R ⁴ , R ⁵ are independently selected from H, -C(O)OR ^x , -C( O) ^Rx , -OC ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl or 4-7 membered heterocyclyl, the -OC ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl or 4-7 membered heterocyclyl is optionally substituted by R ^x .
3. The compound represented by formula (I) or a pharmaceutically acceptable salt according to claim 2, characterized in that: R ³ , R ⁴ , and R ⁵ are independently selected from H, C ₁ -C ₆ alkyl, C ₃ - C ₁₀ cycloalkyl or 4-7 membered heterocyclyl, the C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl or 4-7 membered heterocyclyl is optionally substituted by R ^x .
4. The compound represented by formula (I) or a pharmaceutically acceptable salt according to any one of claims 1-3, characterized in that: Ring A is selected from phenyl or 5-6 membered heteroaryl, and the phenyl or 5-6 membered heteroaryl optionally substituted with one or more ^Ra .
5. The compound represented by formula (I) or a pharmaceutically acceptable salt according to any one of claims 1-4, characterized in that: Ring B is selected from C ₆ -C ₁₀ aryl or 5-10 membered heteroaryl , the C ₆ -C ₁₀ aryl group or 5-10 membered heteroaryl group is optionally substituted by one or more R ^b .
6. The compound represented by formula (I) or a pharmaceutically acceptable salt according to claim 5, characterized in that: Ring B is selected from C ₆ -C ₁₀ aryl groups, and the C ₆ -C ₁₀ aryl groups are optionally replaced by One or more R ^b are substituted.
7. The compound represented by formula (I) or a pharmaceutically acceptable salt according to any one of claims 1 to 6, characterized in that ring C is selected from a 5-10 membered heteroaryl group, and the 5-10 membered heteroaryl group Aryl is optionally substituted with one or more ^Rc .
8. The compound represented by formula (I) or a pharmaceutically acceptable salt according to any one of claims 1-7, characterized in that: each R ^a , R ^b , R ^c is independently selected from halogen, C ₁ - C ₆ alkyl, -OC ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl or 4-10 membered heterocyclyl, the C ₁ -C ₆ alkyl, -OC ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl or 4-10 membered heterocyclyl is optionally substituted by ^Rx .
9. The compound represented by formula (I) or a pharmaceutically acceptable salt according to any one of claims 1-8, characterized in that: R ¹ and R ² are independently selected from H, halogen, C ₁ -C ₆ alkane or C ₃ -C ₁₀ cycloalkyl, which C ₁ -C ₆ alkyl or C ₃ -C ₁₀ cycloalkyl is optionally substituted by R ^x .
10. The compound represented by formula (I) or a pharmaceutically acceptable salt according to any one of claims 1-9, characterized in that: R ^x is selected from halogen, NH ₂ or C ₁ -C ₆ alkyl, said NH ₂ or C ₁ -C ₆ alkyl is optionally substituted by C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl or 4-7 membered heterocyclyl.
11. The compound represented by formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: the compound represented by formula (I) or a pharmaceutically acceptable salt thereof is selected from the group consisting of compounds represented by formula (II) or its pharmaceutically acceptable salt,
    

    Ring A, Ring B, Ring C, R ¹ , R ² , R ⁴ and R ⁵ are as defined in claim 1.
12. The compound represented by formula (II) or a pharmaceutically acceptable salt according to claim 11, characterized in that: R ⁴ is selected from H, C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl or 4- 7-membered heterocyclyl group, the C ₁ -C ₆ alkyl group, C ₃ -C ₁₀ cycloalkyl group or 4-7 membered heterocyclyl group is optionally substituted by R ^x .
13. The compound represented by formula (II) or a pharmaceutically acceptable salt according to claim 11 or 12, characterized in that: R ⁵ is selected from H or C ₁ -C ₆ alkyl.
14. The compound represented by formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: the compound represented by formula (I) or a pharmaceutically acceptable salt thereof is selected from the group consisting of compounds represented by formula (III) or its pharmaceutically acceptable salt,
    

    in, Represents a single or double bond;

    Ring A, ring B, ring C, R ¹ , R ² , X, and Z are as defined in claim 1.
15. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: the compound of formula (I) or a pharmaceutically acceptable salt thereof is selected from the following compounds or a pharmaceutically acceptable salt thereof ,
    
    
    
    
16. A pharmaceutical composition comprising the compound of formula (I) according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof, and pharmaceutically acceptable excipients.
17. The use of the compound of formula (I) or pharmaceutically acceptable salt according to any one of claims 1 to 15, or the pharmaceutical composition according to claim 16 in the preparation of drugs for preventing or treating diseases mediated by USP1.
18. A method of treating a USP1-mediated disease in a mammal, comprising administering a therapeutically effective amount of a compound of formula (I) according to any one of claims 1-15 or a pharmaceutical agent thereof to a mammal in need of the treatment, preferably a human. an acceptable salt, or a pharmaceutical composition according to claim 16.