Tricyclic compounds and their applications

Abstract

The present disclosure describes compounds having USP1 inhibitory activity or function and pharmaceutically acceptable salts thereof. The present disclosure also describes pharmaceutical compositions comprising the compounds of formula (I) or pharmaceutically acceptable salts thereof, and uses thereof as USP1 inhibitors in preventing or treating related diseases.

Description

Tricyclic compounds and their applications

[0001] This disclosure claims priority to a prior application filed with the State Intellectual Property Office of China on August 26, 2022, with patent application number 202211035206.9, entitled “Tricyclic compounds and their applications.” The entire text of the prior application is incorporated herein by reference. Technical Field

[0002] The present disclosure belongs to the field of medical technology, and specifically relates to a class of tricyclic compounds, or pharmaceutically acceptable salts thereof, pharmaceutical compositions containing them, and their use as ubiquitin-specific protease 1 (USP1) inhibitors in preventing or treating diseases associated with USP1. Background Art

[0003] Ubiquitination is a reversible process that involves a series of deubiquitinating enzymes (DUBs) that regulate various cellular processes by deubiquitinating substrates. DUBs are encoded by approximately 100 human genes and are divided into 6 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, a phenomenon that supports the hypothesis that targeting specific DUB enzymes can increase the ubiquitination and degradation of oncogenic substrates and regulate the activity of other key proteins involved in tumor growth, survival, differentiation, and tumor microenvironment maintenance (Hussain, S., et.al., "DUBs and cancer: The role of deubiquitinating enzymes as oncogenes, non-oncogenes and tumor suppressors." Cell Cycle 8, 1688-1697 (2009)).

[0004] Ubiquitin-specific protease 1 (USP1) is a cysteine ​​isopeptidase of the USP subfamily of DUBs. The full-length human USP1 consists of 785 amino acids, including a catalytic triad consisting of Cys90, His593 and Asp751 (Nijman, SMB, 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 can only obtain full enzymatic activity when it combines with the auxiliary factor UAF1 to form a complex required for deubiquitinase activity. The USP1 / UAF1 complex deubiquitinates monoubiquitinated PCNA (proliferating cell nuclear antigen) and monoubiquitinated FANCD2 (Fanconi anemia group complementary group USP1 is a key protein in the repair of DNA damage caused by DNA crosslinking agents such as cisplatin and mitomycin C (MMC). The USP1 / UAF1 complex also deubiquitinates FANCI (Fanconi anemia complementation group 1). 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 to inhibit DNA repair can induce apoptosis in multiple myeloma cells and enhance the sensitivity of lung cancer cells to cisplatin. These findings suggest that USP1 is a promising target for chemotherapy of certain cancers.

[0005] In summary, targeted inhibition of USP1 protein is a potential approach to prevent or treat cancer and other diseases. Therefore, the development of small molecule inhibitors of USP1 is necessary.

[0006] Summary of the Invention

[0007] In one aspect, the present disclosure relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof,

[0008] in,

[0009] X 1 、X 2 、X 3 、X 4 、X 5 、X 6Independently selected from CR 5 or N;

[0010] R 1 、R 2 independently selected from H, halogen, CN, OH, NH2, C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-10 membered heterocyclic group, the OH, NH2, C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-10 membered heterocyclic group is optionally replaced by R x replace;

[0011] or R 1 、R 2 and the atoms they are connected to form C3-C 10 Cycloalkyl or 4-7 membered heterocyclic group, the C3-C 10 Cycloalkyl or 4-7 membered heterocyclic group is optionally replaced by R x replace;

[0012] R 3 、R 4 、R 5 independently selected from H, halogen, CN, OH, NH2, -C(O)OR x 、-C(O)R x , C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C 10 Cycloalkyl, 4-7 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl, the OH, NH2, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C 10 Cycloalkyl, 4-7 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl is optionally replaced by R x replace;

[0013] Ring A is selected from C6-C 10 Aryl or 5-10 membered heteroaryl, the C6-C 10 Aryl or 5-10 membered heteroaryl is optionally substituted by one or more R a replace;

[0014] Ring B is selected from C6-C 10 Arylene, 5-10 membered heteroarylene, 4-10 membered heterocyclylene, C4-C 10 Cycloalkenylene or C3-C 10 Cycloalkylene, the C6-C 10 Arylene, 5-10 membered heteroarylene, 4-10 membered heterocyclylene, C4-C 10 Cycloalkenylene or C3-C 10Cycloalkylene is optionally substituted with one or more R b replace;

[0015] Ring C is selected from C3-C 10 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl, the C3-C 10 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl is optionally substituted by one or more R c replace;

[0016] Every R a 、R b 、R c independently selected from halogen, CN, OH, NH2, -C(O)OR x , C1-C6 alkyl, -O-C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-10 membered heterocyclic group, the NH2, C1-C6 alkyl, -O-C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-10 membered heterocyclic group is optionally replaced by R x replace;

[0017] or R b 、R c and the atoms they are connected to form C4-C 10 Cycloalkenyl or 4-10 membered heterocyclic group, the C4-C 10 Cycloalkenyl or 4-10 membered heterocyclic group is optionally replaced by R x replace;

[0018] R x Selected from halogen, CN, OH, NH2, C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-8 membered heterocyclic group, the OH, NH2, C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-8 membered heterocyclic group is optionally replaced by R d replace;

[0019] R d Selected from halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl or 4-8 membered heterocyclyl.

[0020] In some embodiments, X 5 、X 6 All are selected from N.

[0021] In some embodiments, X 1 、X 2 、X 4 、X 5 、X6 Independently selected from N, X 3 Selected from CR 5 .

[0022] In some embodiments, X 1 、X 2 、X 4 、X 5 、X 6 Independently selected from N, X 3 Selected from CH.

[0023] In some embodiments, R 1 、R 2 independently selected from H, halogen, C1-C6 alkyl or C3-C 10 Cycloalkyl, the C1-C6 alkyl or C3-C 10 The cycloalkyl group is optionally replaced by R x replace.

[0024] In some embodiments, R 1 and R 2 Both are H.

[0025] In some embodiments, R 3 、R 4 、R 5 independently selected from H, halogen, CN, OH, NH2, C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-7 membered heterocyclic group, the OH, NH2, C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-7 membered heterocyclic group is optionally replaced by R x replace.

[0026] In some embodiments, R 3 、R 4 、R 5 Independently selected from H, C1-C6 alkyl or C3-C 10 Cycloalkyl, the C1-C6 alkyl or C3-C 10 The cycloalkyl group is optionally replaced by R x replace.

[0027] In some embodiments, R 3 、R 4 、R 5 Independently selected from H, CH3, CH2CH3, CH(CH3)2 or cyclopropyl.

[0028] In some embodiments, R 3 Selected from H, C1-C6 alkyl or C3-C 10 Cycloalkyl, and R 4 Selected from H or C1-C6 alkyl.

[0029] In some embodiments, R 3 Selected from H, C1-C6 alkyl or C3-C 10 Cycloalkyl, and R 4 Selected from H.

[0030] In some embodiments, R 3 is selected from H, C1-C6 alkyl, and R 4 Selected from H.

[0031] In some embodiments, R 5 is selected from H or C1-C6 alkyl, the C1-C6 alkyl is optionally replaced by R x replace.

[0032] In some embodiments, R 5 Selected from H.

[0033] In some embodiments, ring A is selected from phenyl or 5-6 membered heteroaryl, wherein the phenyl or 5-6 membered heteroaryl is optionally substituted with one or more R a replace.

[0034] In some embodiments, Ring A is selected from phenyl, pyridinyl, or pyrimidinyl, optionally substituted by one or more R a replace.

[0035] In some embodiments, Ring A is selected from pyrimidinyl, optionally substituted with one or more R a replace.

[0036] In some embodiments, Ring B is selected from C6-C 10 Arylene or 5-10 membered heteroarylene, the C6-C 10 Arylene or 5-10 membered heteroarylene is optionally substituted by one or more R b replace.

[0037] In some embodiments, Ring B is selected from C6-C 10 Arylene, the C6-C 10 The arylene group is optionally substituted by one or more R b replace.

[0038] In some embodiments, Ring B is selected from phenylene, optionally substituted with one or more R b replace.

[0039] In some embodiments, Ring C is selected from C6-C 10 Aryl or 5-10 membered heteroaryl, the C6-C 10 Aryl or 5-10 membered heteroaryl is optionally substituted by one or more Rc replace.

[0040] In some embodiments, Ring C is selected from 5-10 membered heteroaryl, wherein the 5-10 membered heteroaryl is optionally substituted with one or more R c replace.

[0041] In some embodiments, Ring C is selected from 5-6 membered heteroaryl, said 5-6 membered heteroaryl being optionally substituted with one or more R c replace.

[0042] In some embodiments, Ring C is selected from pyrazolyl or imidazolyl, which is optionally substituted with one or more R c replace.

[0043] In some embodiments, each R a 、R b 、R c independently selected from halogen, C1-C6 alkyl, -O-C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-10 membered heterocyclic group, the C1-C6 alkyl, -O-C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-10 membered heterocyclic group is optionally replaced by R x replace.

[0044] In some embodiments, each R a 、R b 、R c independently selected from halogen, C1-C6 alkyl, -O-C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-6 membered heterocyclic group, wherein the C1-C6 alkyl, -O-C1-C6 alkyl, C3-C6 cycloalkyl or 4-6 membered heterocyclic group is optionally replaced by R x replace.

[0045] In some embodiments, each R a Independently selected from C1-C6 alkyl, -O-C1-C6 alkyl or C3-C 10 Cycloalkyl, the C1-C6 alkyl, -O-C1-C6 alkyl or C3-C 10 The cycloalkyl group is optionally replaced by R x replace.

[0046] In some embodiments, each R c Independently selected from C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-6 membered heterocyclic group, the C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-6 membered heterocyclic group is optionally replaced by R x replace.

[0047] In some embodiments, each R a 、R b 、R c independently selected from halogen, CH3, -CH(CH3)2, -CH2CH3, -O-CH3, cyclopropyl, oxetanyl or azetidinyl, wherein the CH3, -CH(CH3)2, -CH2CH3, -O-CH3, cyclopropyl, oxetanyl or azetidinyl is optionally replaced by R x replace.

[0048] In some embodiments, each R a independently selected from -O-CH3 or cyclopropyl, said -O-CH3 or cyclopropyl being optionally replaced by R x replace.

[0049] In some embodiments, each R c independently selected from -CH3, -CH(CH3)2, -CH2CH3, cyclopropyl, oxetanyl or azetidinyl, wherein the -CH3, -CH(CH3)2, -CH2CH3, cyclopropyl, oxetanyl or azetidinyl is optionally replaced by R x replace.

[0050] In some embodiments, each R a 、R b 、R c Independently selected from F, CH3, -CH(CH3)2, -CH2CH3, -CHF2, -CF3, -O-CH3, -O-CHF2, cyclopropyl, oxetanyl or

[0051] In some embodiments, R b 、R c and the atoms to which they are attached together form a 4-10 membered heterocyclic group, the 4-10 membered heterocyclic group being optionally replaced by R x replace.

[0052] In some embodiments, R x Selected from halogen, C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-8 membered heterocyclic group, the C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-8 membered heterocyclic group is optionally replaced by R d replace.

[0053] In some embodiments, R x is selected from halogen or C1-C6 alkyl, wherein the C1-C6 alkyl is optionally replaced by R d replace.

[0054] In some embodiments, Rx Selected from F or CH3.

[0055] In some embodiments, the compound represented by formula (I) or a pharmaceutically acceptable salt thereof is selected from the compound represented by formula (II) or a pharmaceutically acceptable salt thereof,

[0056] Among them, ring A, ring B, ring C, R 1 、R 2 、R 3 、R 4 、X 1 、X 2 、X 3 and X 4 As defined above.

[0057] In the absence of conflict, it should be understood that the above embodiments can be arbitrarily combined to form a technical solution including the features of the combined embodiments. Such a combined technical solution is within the scope of the present invention.

[0058] 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,

[0059] In another aspect, the present disclosure provides a pharmaceutical composition comprising a compound of formula (I) or formula (II) of the present disclosure or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0060] In another aspect, the present disclosure provides a method for treating a disease mediated by USP1 in a mammal, comprising administering a therapeutically effective amount of a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof to a mammal, preferably a human, in need of such treatment.

[0061] In another aspect, the present disclosure provides use of a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the preparation of a medicament for preventing or treating a disease mediated by USP1.

[0062] In another aspect, the present disclosure provides use of a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in preventing or treating a disease mediated by USP1.

[0063] 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 a disease mediated by USP1.

[0064] In some embodiments, the USP1 -mediated disease is cancer.

[0065] Definitions and Explanations of Terms

[0066] Unless otherwise indicated, the terms used in this disclosure have the following meanings. The definitions of groups and terms described in this disclosure, including their definitions as examples, exemplary definitions, preferred definitions, definitions described in tables, and definitions of specific compounds in the examples, may be combined and coupled with each other in any manner. A particular term should not be considered as undefined or unclear unless specifically defined, but should be understood according to its ordinary meaning in the art. When a trade name appears in this document, it is intended to refer to the corresponding commercial product or its active ingredient.

[0067] In this article Indicates the attachment site.

[0068] In this article, multiple arrows in the synthetic route Indicates a multi-step reaction.

[0069] The diagrammatic representations of racemates or enantiomerically pure compounds herein are from Maehr, J. Chem. Ed. 1985, 62: 114-120. Unless otherwise indicated, wedge and dotted wedge keys are used. To indicate the absolute configuration of a stereocenter, use black real and imaginary bonds. Indicates the relative configuration of a stereocenter (such as the cis-trans configuration of an alicyclic compound).

[0070] The term "tautomer" refers to functional group isomers resulting from the rapid shift of an atom in a molecule between two positions. The compounds of the present disclosure may exhibit tautomerism. Tautomeric compounds may exist as two or more interconvertible species. Tautomers generally exist in equilibrium, and attempts to isolate a single tautomer usually result in a mixture whose physical and chemical properties are consistent with a 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 predominates, while in phenols, the enol form predominates. The present disclosure encompasses all tautomeric forms of the compounds.

[0071] The term "stereoisomer" refers to isomers resulting from different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers and diastereomers.

[0072] The compounds of the present invention may have asymmetric atoms such as carbon atoms, sulfur atoms, nitrogen atoms, phosphorus atoms or asymmetric double bonds, so that the compounds of the present invention 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 mixtures or other mixtures thereof, such as mixtures enriched in enantiomers or diastereomers, all of which are within the definition of the compounds of the present invention and mixtures thereof. Additional asymmetric carbon atoms, asymmetric sulfur atoms, asymmetric nitrogen atoms or asymmetric phosphorus atoms may be present in substituents such as alkyl groups, and all of which are within the definition of the compounds of the present invention and mixtures thereof. Compounds of the present disclosure containing an asymmetric atom can be isolated in optically pure or racemic forms. Optically pure forms can be resolved from racemic mixtures or synthesized by using chiral starting materials or reagents.

[0073] The term "substituted" refers to the replacement of any one or more hydrogen atoms on a particular atom by a substituent, as long as the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is an oxo (i.e., =O), it means that two hydrogen atoms are replaced. Oxo does not occur on aromatic groups.

[0074] The term "optional" or "optionally" refers to that the event or situation described subsequently may or may not occur, and the description includes that the event or situation occurs and that the event or situation does not occur. For example, an ethyl group is "optionally" substituted with halogen, meaning that the ethyl group may be unsubstituted (CH2CH3), monosubstituted (CH2CH2F, CH2CH2Cl, etc.), polysubstituted (CHFCH2F, CH2CHF2, CHFCH2Cl, CH2CHCl2, etc.), or fully substituted (CF2CF3, CF2CCl3, CCl2CCl3, etc.). It will be appreciated by those skilled in the art that for any group comprising one or more substituents, any sterically impossible and / or incomposable replacement or substitution pattern will not be introduced.

[0075] When any variable (such as R a 、R b ) appears more than once in the composition or structure of a compound, its definition is independent in each case. For example, if a group is represented by two R b is replaced, then each R b There are independent options.

[0076] When the linking group mentioned in this article does not specify its connection direction, its connection direction is arbitrary. L in 1 When selected from "C1-C3 alkylene-O", L 1 You can connect rings Q and R from left to right. 1 Constitute "ring Q-C1-C3 alkylene-OR 1 ", you can also connect rings Q and R from right to left 1 Constitute the "ring QO-C1-C3 alkylene-R 1 ”.

[0077] When a substituent's bond crosses two atoms in a ring, the substituent may be bonded to any atom in the ring. Represents R 5 Substitution can occur at any position on the benzene ring.

[0078] In this article, C m -C n It means having an integer number of carbon atoms in the range mn. For example, "C1-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.

[0079] The term "alkyl" refers to a group of the formula C n H 2n+1 The term "C1-C 10The term "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 radical 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-dimethylbutyl , 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, etc.; the term "C1-C6 alkyl" may be understood to mean an alkyl group having 1, 2, 3, 4, 5 or 6 carbon atoms, specific examples of which 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 "C1-C3 alkyl" may be understood to mean a straight-chain or branched saturated alkyl group having 1, 2 or 3 carbon atoms. The "C1-C 10 The term "alkyl" may include "C1-C6 alkyl" or "C1-C3 alkyl" and the like, and the "C1-C6 alkyl" may further include "C1-C3 alkyl".

[0080] The term "alkenyl" refers to a linear or branched unsaturated aliphatic hydrocarbon group consisting of carbon atoms and hydrogen atoms and having at least one double bond. 10 "Alkenyl" is understood to mean a linear 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, "C2-C 10 "Alkenyl" is preferably "C2-C6 alkenyl", further preferably "C2-C4 alkenyl", and further preferably C2 or C3 alkenyl. It will be understood that when the alkenyl contains more than one double bond, the double bonds may be separated from each other or conjugated. Specific examples of the alkenyl 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, etc.

[0081] The term "alkynyl" refers to a straight or branched unsaturated aliphatic hydrocarbon group consisting of carbon atoms and hydrogen atoms and having at least one triple bond. 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. "C2-C 10 Examples of "alkynyl" include, but are not limited to, ethynyl (-C≡CH), propynyl (-C≡CCH 3、 -CH2C≡CH), but-1-ynyl, but-2-ynyl or but-3-ynyl. "C2-C 10 The term "alkynyl" may include "C2-C3 alkynyl". Examples of "C2-C3 alkynyl" include ethynyl (-C≡CH), prop-1-ynyl (-C≡CCH3), and prop-2-ynyl (-CH2C≡CH).

[0082] The term "cycloalkyl" refers to a fully saturated carbon ring that exists in the form of a monocyclic, fused, bridged, or spirocyclic ring. Unless otherwise indicated, the carbon ring is typically a 3- to 10-membered ring. The term "C3-C 10 "Cycloalkyl" is understood to mean a saturated monocyclic, bicyclic, spirocyclic or bridged ring having 3, 4, 5, 6, 7, 8, 9 or 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, and the like. The term "C3-C 10 "Cycloalkyl" may include "C3-C6 cycloalkyl". The term "C3-C6 cycloalkyl" may be understood to mean a saturated monocyclic or bicyclic hydrocarbon ring having 3, 4, 5 or 6 carbon atoms. Specific examples include but are not limited to cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

[0083] The term "cycloalkylene" refers to a radical having two hydrogen atoms derived from the same carbon atom or two different carbon atoms of a parent cycloalkane, wherein cycloalkyl is as defined above.

[0084] The term "cycloalkenyl" refers to a non-aromatic carbocyclic ring that is not fully saturated and exists in the form of a monocyclic, fused, bridged, or spirocyclic ring. Unless otherwise indicated, the carbocyclic ring is typically a 5- to 8-membered ring. Specific examples of the cycloalkenyl group include, but are not limited to, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, cycloheptenyl, or cycloheptadienyl.

[0085] The term "cycloalkenylene" refers to a radical having two hydrogen atoms derived from the same carbon atom or two different carbon atoms of a parent cycloalkene, wherein cycloalkenyl is as defined above.

[0086] The term "heterocyclyl" refers to a fully saturated or partially saturated (heteroaromatic group that is not aromatic as a whole) monocyclic, fused, spirocyclic or bridged ring group, which contains 1 to 5 heteroatoms or heteroatomic groups (i.e., heteroatom-containing groups) in its ring atoms, wherein the "heteroatoms or heteroatomic groups" include, but are 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-. The term "3-10 membered heterocyclyl" refers to a heterocyclyl group having 3, 4, 5, 6, 7, 8, 9 or 10 ring atoms, and containing 1 to 5 heteroatoms or heteroatomic groups independently selected from the above-mentioned heteroatoms in its ring atoms. “3-10 membered heterocyclic group” includes “4-7 membered heterocyclic group”, wherein specific examples of 4 membered heterocyclic group include but are not limited to azetidinyl or oxetanyl; specific examples of 5 membered heterocyclic group include but are not limited to tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl, 4,5-dihydrooxazolyl or 2,5-dihydro-1H-pyrrolyl; specific examples of 6 membered heterocyclic group include but are not limited to tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, trithianyl, tetrahydropyridinyl or 4H-[1,3,4]thiadiazinyl; specific examples of 7 membered heterocyclic group 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]pyrrol-2(1H)-yl; specific examples of 5,6-membered bicyclic groups include but are not limited to hexahydropyrrolo[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 ring group of the above-mentioned 4-7-membered heterocyclic groups, specific examples of which include but are not limited to dihydroisoquinolinyl and the like. The term "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", etc. The term "4-7 membered heterocyclyl" may further include "4-6 membered heterocyclyl", "5-6 membered heterocyclyl", "4-7 membered heterocycloalkyl", "4-6 membered heterocycloalkyl", "5-6 membered heterocycloalkyl", etc. Although some bicyclic heterocyclyl groups in the present disclosure partially contain a benzene ring or a heteroaromatic ring, the heterocyclyl group as a whole is still non-aromatic.

[0087] The term "heterocyclylene" refers to a radical having two hydrogen atoms derived from the same carbon atom or two different carbon atoms of a parent heterocyclic ring, wherein heterocyclyl is as defined above.

[0088] The term "aryl" refers to an all-carbon monocyclic or fused polycyclic aromatic ring group having a conjugated π electron system. The aryl group may have 6-20 carbon atoms, 6-14 carbon atoms, or 6-12 carbon atoms. The term "C6-C 20 "Aryl" is understood to be an aromatic radical having 6 to 20 carbon atoms. In particular, a ring having 6 carbon atoms ("C6 aryl"), such as phenyl; or a ring having 9 carbon atoms ("C9 aryl"), such as indanyl or indenyl; or a ring having 10 carbon atoms ("C 10 or a ring having 13 carbon atoms ("C 13 aryl) such as fluorenyl; or a ring having 14 carbon atoms ("C 14 The term "C6-C 10 "Aryl" is understood to be an aromatic radical having 6 to 10 carbon atoms. In particular, a ring having 6 carbon atoms ("C6 aryl"), such as phenyl; or a ring having 9 carbon atoms ("C9 aryl"), such as indanyl or indenyl; or a ring having 10 carbon atoms ("C 10 The term "C6-C 20 "Aryl" may contain "C6-C 10 Aryl".

[0089] The term "arylene" refers to a radical having two hydrogen atoms derived from the same carbon atom or two different carbon atoms of a parent aromatic ring, wherein aryl is as defined above.

[0090] The term "heteroaryl" refers to a monocyclic or fused polycyclic ring system with aromatic character, which contains at least one ring atom selected from N, O, 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 containing 1 to 5, preferably 1 to 3 heteroatoms independently selected from N, O and S. In particular, the heteroaryl group is selected from thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl or thiadiazolyl, and the like, and benzo derivatives thereof, such as benzofuranyl, benzothienyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzotriazolyl, indazolyl, indolyl or isoindolyl, and the like; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl or triazinyl, and the like, and benzo derivatives thereof, such as quinolyl, quinazolinyl or isoquinolyl, and the like; or acinyl, indolizinyl, purinyl, and the like, and benzo derivatives thereof; or cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl or phenoxazinyl, and the like. 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.

[0091] The term "heteroarylene" refers to a radical having two hydrogen atoms derived from the same carbon atom or two different carbon atoms of a parent heteroaryl ring, wherein heteroaryl is as defined above.

[0092] The term "halo" or "halogen" refers to fluorine, chlorine, bromine or iodine.

[0093] The term "therapeutically effective amount" means:

[0094] An amount of a compound of the present disclosure that (i) treats a particular disease, condition, or disorder, (ii) alleviates, ameliorates, or eliminates one or more symptoms of a particular disease, condition, or disorder, or (iii) delays the onset of one or more symptoms of a particular disease, condition, or disorder as described herein.

[0095] The amount of a compound of the disclosure that constitutes a "therapeutically effective amount" varies 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 their own knowledge and this disclosure.

[0096] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0097] The term "pharmaceutically acceptable salt" refers to a salt of a pharmaceutically acceptable acid or base, including a salt formed between a compound and an inorganic acid or organic acid, and a salt formed between a compound and an inorganic base or an organic base.

[0098] The term "pharmaceutical composition" refers to a mixture of one or more compounds of the present disclosure or their salts and a pharmaceutically acceptable excipient. The purpose of a pharmaceutical composition is to facilitate administration of the compounds of the present disclosure to an organism.

[0099] The term "pharmaceutically acceptable excipient" refers to an excipient that is non-irritating to organisms and does not impair the biological activity and properties of the active compound. Suitable excipients are well known to those skilled in the art and include, for example, carbohydrates, waxes, water-soluble and / or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, and the like.

[0100] Examples of mammals include, but are not limited to, any member of the class Mammalia: humans, non-human primates (e.g., chimpanzees and other apes and monkeys); livestock, such as cattle, horses, sheep, goats, pigs; domestic animals, such as rabbits, dogs, and cats; laboratory animals, including rodents, such as rats, mice, and guinea pigs. Examples of non-human mammals include, but are not limited to, birds and fish. In one embodiment of the methods and compositions provided herein, the mammal is a human.

[0101] The word "comprise" or "comprises" and its English variations such as comprises or comprising are to be understood as having an open and non-exclusive meaning, ie, "including but not limited to".

[0102] The present disclosure also includes isotopically labeled compounds of the present disclosure that are identical to those described herein, but where one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can 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 N. 15 N. 15 O.17 O. 18 O. 31 P. 32 P. 35 S. 18 F. 123 I. 125 I and 36 Cl et al.

[0103] Certain isotopically labeled compounds of the present disclosure (e.g., 3 H and 14 C-labeled) can be used in compound and / or substrate tissue distribution assays. 3 H) and carbon-14 (i.e. 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. Isotopically labeled compounds of the disclosure can generally be prepared by following procedures analogous to those disclosed in the Schemes and / or Examples below, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

[0104] The pharmaceutical compositions of the present disclosure can be prepared by combining the compounds of the present disclosure with suitable pharmaceutically acceptable excipients, and 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.

[0105] Typical routes of administration of the disclosed compounds, 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.

[0106] The pharmaceutical composition of the present disclosure can be manufactured by methods well known in the art, such as conventional mixing methods, dissolution methods, granulation methods, emulsification methods, freeze-drying methods, and the like.

[0107] In some embodiments, the pharmaceutical composition is in oral form. For oral administration, the pharmaceutical composition can be formulated by mixing the active compound with pharmaceutically acceptable excipients well known in the art. These excipients enable the compounds of the present disclosure to be formulated into tablets, pills, lozenges, dragees, capsules, liquids, gels, slurries, suspensions, and the like for oral administration to a patient.

[0108] Solid oral compositions can be prepared by conventional mixing, filling, or tableting methods. For example, they can be prepared by mixing the active compound with a solid excipient, optionally grinding the resulting mixture, adding other suitable excipients as needed, and then granulating the mixture to obtain a tablet or dragee core. Suitable excipients include, but are not limited to, binders, diluents, disintegrants, lubricants, glidants, or flavoring agents.

[0109] The pharmaceutical composition may also be suitable for parenteral administration, such as sterile solutions, suspensions or lyophilized products in appropriate unit dosage forms.

[0110] In all methods of administration of the compounds of formula (I) described herein, the dosage administered per day is 0.01 mg / kg to 200 mg / kg body weight, in the form of single or divided doses. DETAILED DESCRIPTION

[0111] The present invention is described in detail below with reference to the examples, but the following examples should not be construed as limiting the scope of the present invention.

[0112] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). The units of NMR shifts are 10 -6 (ppm). The solvents for NMR determination are deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, etc., and the internal standard is tetramethylsilane (TMS); "IC 50 ” refers to the half-maximal inhibitory concentration, which is the concentration at which half of the maximum inhibitory effect is achieved.

[0113] Unless otherwise specified, the ratios expressed for mixed solvents are volume ratios.

[0114] Unless otherwise stated, % refers to wt%.

[0115] Compounds are manually or Software naming, commercially available compounds use supplier catalog names.

[0116] The eluent mentioned below can be a mixed eluent formed by two or more solvents, and the ratio is the volume ratio of each solvent, such as petroleum ether:ethyl acetate = 5:1-1:1, which means that during the elution process, the volume ratio of petroleum ether to ethyl acetate in the mixed eluent is 5:1-1:1.

[0117] Abbreviations:

[0118] THF: tetrahydrofuran; DIPEA: N,N-diisopropylethylamine; Ti(OiPr)4: tetraisopropyl titanate; PE: petroleum ether; EA or EtOAc: ethyl acetate; NaBH4: sodium borohydride; Xphos Pd G2: chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II); dioxane: dioxane; DCM: dichloromethane.

[0119] Example 1: 8-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-10-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-5,10-dihydropyrimidin[4,5-d][1,2,4]triazolo[4,3-a]pyrimidine (Compound 1)

[0120] Step 1: Methyl 2-chloro-4-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)amino)pyrimidine-5-carboxylate (1c)

[0121] Methyl 2,4-dichloropyrimidine-5-carboxylate (4.0 g, 19.3 mmol) was dissolved in tetrahydrofuran (75 mL) and cooled to 0°C. A solution of compound 1b (4.9 g, 19.3 mmol) and N,N-diisopropylethylamine (7.5 g, 58.0 mmol, 10.1 mL) in tetrahydrofuran (15 mL) was slowly added dropwise to the reaction mixture with stirring. The reaction mixture was allowed to react at 0°C for 3 h, then quenched with saturated brine (100 mL) and extracted three times with 50 mL of ethyl acetate. The combined organic phases were washed with saturated brine (50 mL), and the solvent was removed by distillation under reduced pressure. The resulting residue was purified on a silica gel column (petroleum ether:ethyl acetate = 5:1-1:1) to obtain compound 1c (5.6 g, yield 68%). m / z (ESI): 426.1 [M+H] + .

[0122] Step 2: 2-Chloro-4-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)amino)pyrimidin-5-yl)methanol (1d)

[0123] Dissolve 1c (2.7 g, 6.3 mmol) in tetrahydrofuran (10 mL) and slowly add lithium aluminum hydride (1 M, 9.5 mL) in an ice-salt bath at -10°C. The resulting reaction mixture is allowed to react at -10°C for 2 h. Add 1 mL of water, then 1 mL of 15% aqueous sodium hydroxide solution, and then 3 mL of water. Stir at room temperature for 0.5 h, filter, rinse the filter cake with ethyl acetate, and concentrate the combined filtrates to yield 1d (2.6 g crude product). m / z (ESI): 398.1 [M+H] + .

[0124] Step 3: 2-Chloro-4-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)amino)pyrimidine-5-carbaldehyde (1e)

[0125] Dissolve 1d (2.5 g, 6.3 mmol) in dichloromethane (80 mL) and add manganese dioxide (11.2 g, 125.7 mmol). The resulting mixture is reacted at 10°C for 2 h. The reaction mixture is filtered through celite, and the filter cake is rinsed with ethyl acetate. The filtrate is concentrated to yield 1e (2.1 g, crude product). m / z (ESI): 396.1 [M+H] + .

[0126] Step 4: N-((2-chloro-4-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)amino)pyrimidin-5-yl)methylene)-2-methylpropane-2-sulfenamide (1 g)

[0127] 1e (2.0 g, 5.1 mmol), 2-methyl-2-propanesulfenamide 1f (918.7 mg, 7.6 mmol), and tetraisopropyl titanate (4.3 g, 15.2 mmol, 4.5 mL) were dissolved in tetrahydrofuran (20 mL). The resulting reaction solution was stirred at 50°C for 16 h. The reaction solution was diluted with 20 mL of methanol, followed by 5 mL of saturated aqueous sodium bicarbonate. The resulting mixture was stirred at room temperature for 0.5 h, then filtered, and the filter cake was rinsed with ethyl acetate. The combined filtrates were then distilled under reduced pressure to remove the solvent. The resulting residue was purified on a silica gel column (PE:EA = 3:1 to 0:1) to obtain compound 1g (2.5 g, 99% yield). m / z (ESI): 499.1 [M+H] + .

[0128] Step 5: N-((2-chloro-4-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)amino)pyrimidin-5-yl)methyl)-2-methylpropane-2-sulfenamide (1h)

[0129] Dissolve 1g (2.4g, 4.8mmol) in tetrahydrofuran (10mL), stir at 0°C, and slowly add sodium borohydride (273.0mg, 7.2mmol). The resulting reaction solution was stirred at 0°C for 2h. The reaction solution was quenched with 30mL of water and extracted with ethyl acetate (20mL*3). The resulting organic phases were combined and washed with saturated brine (20mL), and the solvent was removed by distillation under reduced pressure. The resulting residue was purified on a silica gel column (PE:EA = 2:1-0:1) to obtain compound 1h (1.3g, yield 54%). m / z (ESI): 501.1 [M+H] + .

[0130] Step 6: N-((4'-cyclopropyl-6'-methoxy-4-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)amino)-[2,5'-bipyrimidinyl]-5-yl)methyl)-2-methylpropane-2-sulfenamide (1j)

[0131] 1h (1.0 g, 2.0 mmol), (4-cyclopropyl-6-methoxypyrimidin-5-yl)boronic acid 1i (774.5 mg, 4.0 mmol), chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (314.1 mg, 399.2 μmol), and potassium phosphate (1.3 g, 6.0 mmol) were added to water (0.8 mL) and dioxane (8 mL). The mixture was stirred at 100°C under nitrogen for 2 h. The reaction mixture was concentrated and the resulting residue was purified on a silica gel column (PE:EA = 2:1 to 0:1) to afford compound 1j (600 mg, 49% yield). m / z (ESI): 615.2 [M+H] + .

[0132] Step 7: 5-(Aminomethyl)-4'-cyclopropyl-6'-methoxy-N-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-[2,5'-bipyrimidinyl]-4-amine hydrochloride (11)

[0133] 1j (600.0 mg, 976.1 μmol) was added to a 1.0 mol / L hydrochloric acid-ethyl acetate solution (10 mL) and stirred at room temperature for 1 h. The reaction solution was concentrated to give 1l (580 mg crude product). m / z (ESI): 511.2 [M+H] + .

[0134] Step 8: 7-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-3,4-dihydropyrimido[4,5-d]pyrimidine-2(1H)-thione (1n)

[0135] 1l (500.0 mg, 979.4 μmol) and N,N-diisopropylethylamine (1.3 g, 9.8 mmol, 1.7 mL) were dissolved in dichloromethane (8 mL). 1,1'-Thiocarbonyldiimidazole (872.7 mg, 4.9 mmol) was added with stirring, and the mixture was allowed to react at room temperature for 1 h. The reaction solution was quenched with water and extracted with ethyl acetate (20 mL x 3). The combined organic phases were washed with saturated brine (20 mL) and the solvent was removed by distillation under reduced pressure. The resulting residue was purified on a silica gel column (PE:EA = 1:1 to 0:1) to obtain compound 1n (300 mg, 55% yield). m / z (ESI): 553.2 [M+H] + .

[0136] Step 9: 7-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-2-hydrazono-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1,2,3,4-tetrahydropyrimido[4,5-d]pyrimidine (1o)

[0137] 1n (275.0 mg, 497.7 μmol) and hydrazine hydrate (0.1 mL) were added to tetrahydrofuran (3 mL) and reacted at 80°C for 4 h. The reaction solution was concentrated and the resulting crude product was purified by preparative HPLC (Waters AutoP, Xbridge-C18, 19 x 150 mm, gradient elution from 5% to 75% acetonitrile in water, 15 mL / min) and lyophilized to afford 1o (80 mg, 29% yield). m / z (ESI): 551.2 [M+H] + .

[0138] Step 10: 8-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-10-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-5,10-dihydropyrimido[4,5-d][1,2,4]triazolo[4,3-a]pyrimidine (Compound 1)

[0139] 1o (50.0 mg, 90.8 μmol) was dissolved in trimethyl orthoformate (2 mL) and reacted at 100°C for 2 h. The reaction solution was concentrated and purified by preparative HPLC (Waters AutoP, Xbridge-C18, 19*150 mm, 30-65% acetonitrile in water gradient elution, 15 mL / min) and lyophilized to obtain compound 1 (7 mg, 14% yield).

[0140] m / z(ESI):561.1[M+H] + .

[0141] 1 H NMR (400MHz, DMSO-d6) δ8.65(m,2H),8.46(s,1H),7.92(s,1H),7.71–7.60(d,J=8.0Hz,2H),7.51(d,J=8.0 Hz,2H),5.46(s,2H),5.41(s,2H),3.83(s,3H),3.74(s,3H),1.70(m,1H),1.04–0.96(m,2H),0.77(m,2H).

[0142] Example 2: 8-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-10-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-5,10-dihydropyrimido[4,5-d][1,2,4]triazolo[4,3-a]pyrimidine (Compound 2)

[0143] Compound 2 was prepared by replacing 1b in step 1 of Example 1 with 2a and adopting a synthetic route and method similar to that of compound 1.

[0144] m / z(ESI):561.1[M+H] + .

[0145] 1 H NMR (400MHz, DMSO-d6) δ8.70–8.62(m,2H),8.47(s,1H),7.56(d,J=8.2Hz,2H),7.50(d ,J=8.2Hz,2H),6.75(s,1H),5.45(s,2H),5.42(s,2H),3.83(s,3H),2.31(s,3H),1.74– 1.64(m,1H),1.04–0.95(m,2H),0.84–0.72(m,2H).

[0146] Example 3: 8-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-5-methyl-10-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-5,10-dihydropyrimido[4,5-d][1,2,4]triazolo[4,3-a]pyrimidine (Compound 3)

[0147] Step 1: N-(1-(2-chloro-4-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)amino)pyrimidin-5-yl)ethyl)-2-methylpropane-2-sulfenamide (3a)

[0148] Intermediate 1g (1.4g, 2.8mmol) was dissolved in tetrahydrofuran (30mL) and cooled to 0°C. Methylmagnesium bromide (1M, 8.4mL) was added to the reaction solution and allowed to react at 0°C for 2h. The reaction solution was quenched with water and extracted with ethyl acetate (30mL*3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated. The resulting residue was purified on a silica gel column (PE:EA = 2:1-0:1) to obtain compound 3a (0.8g, 55% yield). m / z (ESI): 515.1 [M+H] + .

[0149] Compound 3 was prepared by using a synthetic route and method similar to steps 6-10 of compound 1, replacing 1h in step 6 with 3a.

[0150] m / z(ESI):575.1[M+H] + .

[0151] 1 H NMR (400MHz, DMSO-d6) δ8.76(s,1H),8.65(m,2H),7.92(s,1H),7.68(d,J=8.0Hz,2H),7.49(d,J=8.0Hz,2H),5.83( m,1H),5.42(s,2H),3.83(s,3H),3.74(s,3H),1.85(d,J=6.6Hz,3H),1.75–1.66(m,1H),0.99(m,2H),0.78(m,2H).

[0152] Test Example 1: USP1 enzyme in vitro activity detection experiment

[0153] Experimental instruments:

[0154] Experimental Materials:

[0155] The USP1 enzyme (Recombinant Human His6-USP1 / His6-UAF1 Complex Protein, CF) used in this experiment was purchased from R&D Systems, Cat. No. E-568-050. Store at -80°C after aliquoting.

[0156] The detection kit (Ub-CHOP2-Reporter Deubiquitination Assay Kit) was purchased from LifeSensors, catalog number PR1101. Store at -80°C after aliquoting. The kit contains a ubiquitinated reporter enzyme, which becomes active after deubiquitination by USP1 / UAF1. Upon catalytic conversion of the substrate, the substrate is excited by a 485 nm laser and emits a 531 nm light signal.

[0157] Other reagents and consumables required for the experiment are as follows:

[0158] Experimental methods:

[0159] Test compounds were dissolved in DMSO to 10 mM. Compounds and pure DMSO were dispensed into each well of a 384-well plate using a compound dilution and pipetting instrument (Echo). Starting at a maximum concentration of 3 μM, the compound was diluted three-fold over a total of eight concentration points. 50 nL of test compound or DMSO (as a control) was added to each well. The instrument used different ratios to generate a series of sample dilutions (3000 nM, 1000 nM, 333 nM, 111 nM, 37 nM, 12 nM, 4.1 nM, and 1.4 nM). The enzyme was diluted in freshly prepared reaction solution (20 mM Tris-HCl (pH 8.0), 2 mM CaCl2, 2 mM β-mercaptoethanol, 0.05% CHAPS (CHAPS diluted in ddH2O)). 5 μL of the diluted enzyme reaction solution was added to each well. The enzyme and compound were mixed by centrifugation and vortexing, then centrifuged again and placed on ice. Dilute the reporter system and substrate in the reaction solution, add 5 μL of the diluted solution to each well, and centrifuge to mix. Incubate at room temperature for 0.5 hours. Measure the fluorescence signal in each well using an Envision plate reader (PerkinElmer; excitation wavelength: 485 nm, emission wavelength: 535 nm). The IC value of the compound's inhibitory activity against the enzyme is quantified. 50 The values ​​were calculated using the four-parameter logistic model method. In the following formula, x represents the logarithm of the compound concentration; F(x) represents the effect value (the inhibition rate of enzyme activity under the conditions of this concentration): F(x) = (A + ((BA) / (1 + ((C / x)^D)))). A, B, C, and D are four parameters. IC was calculated using Xlfit. 50 The values ​​were further calculated as the compound concentration required for 50% inhibition of enzyme activity in the best-fit curve.

[0160] The test results are shown in Table 1.

[0161] Table 1 In vitro inhibitory activity of USP1 enzyme

[0162] Test Example 2: Experimental study on the inhibition of MDA-MB-436 cell proliferation by the disclosed compounds

[0163] Experimental instruments:

[0164] Experimental Materials:

[0165] The cells used in the experiment were MDA-MB-436, purchased from Kebai Biotechnology Co., Ltd., catalog number CBP60385. The cells were subcultured in culture medium (DMEM containing 10% FBS) and frozen in liquid nitrogen at low cell passages. The cells used in the experiment did not exceed 15 passages.

[0166] Detection kit ( The Luminescent Cell Viability Assay was purchased from Promega under the catalog number G7573. Store at -30°C after aliquoting. This assay is a homogenous method for determining the number of viable cells in culture by quantifying ATP. The luminescent signal generated by the assay is proportional to the amount of ATP present, which is directly proportional to the number of cells in the culture.

[0167] The information of other reagents and consumables required for the experiment is as follows

[0168] Experimental methods:

[0169] Cultured cells were digested with 0.25% Trypsin-EDTA solution, harvested, centrifuged, and resuspended in culture medium (DMEM supplemented with 10% FBS) at the adjusted concentration. The cells were seeded in 384-well plates (400 cells / 20 μL / well) and cultured overnight in a 37°C, 5% CO2 incubator. Compounds and pure DMSO were dispensed into each well of the 384-well plate using a compound dilution and pipetting system (Echo Acoustic Pipetting System). Starting at a maximum concentration of 10 μM, the compounds were diluted fourfold for a total of eight concentration points. 100 nL of test compound or DMSO (control) was added to each well to obtain a series of sample concentrations (10,000 nM, 2,500 nM, 625 nM, 156 nM, 39 nM, 10 nM, 2.4 nM, and 0.61 nM). 30 μL of culture medium was added to each well, centrifuged and mixed, and then centrifuged again. The cells were then cultured in a cell culture incubator for 7 days (one column of cells was treated with CTG (CellTiter-Glo) assay solution on the day of drug addition). After the 7th day, 25 μL of CTG assay solution was added to each well, centrifuged and mixed, and then incubated at room temperature in the dark for 10 minutes. The chemiluminescence signal in each well was measured using an Envision plate reader (PerkinElmer, emission wavelength 400-700 nm). The chemiluminescence value [RLU]cpd was obtained for the drug-treated group on day 7, and the chemiluminescence value [RLU]cell was obtained for the untreated group treated with DMSO on day 7. In parallel, the chemiluminescence value [RLU]background was obtained for the untreated group treated with DMSO on day 0. The inhibition rate (%) of the compound on proliferation was calculated as [1-([RLU]cpd–[RLU]background) / ([RLU]cell–[RLU]background)] × 100%. The inhibitory activity of the compound on proliferation was calculated as GI. 50 The values ​​were calculated using a four-parameter logistic model. In the following formula, x represents the logarithm of the compound concentration; F(x) represents the effect size (the rate of inhibition of proliferation at that concentration): F(x) = (A + ((BA) / (1 + ((C / x)^D)))). A, B, C, and D are the four parameters. Using Xlfit, GI 50 The values ​​were further calculated as the compound concentration required for 50% inhibition of proliferation in the best-fit curve.

[0170] The inhibitory activity of the disclosed compounds on MDA-MB-436 cell proliferation was determined by the above test. The measured GI 50 See Table 2 for values.

[0171] Table 2 Inhibitory activity of the disclosed compounds on MDA-MB-436 cell proliferation

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof, in, X 1 、X 2 、X 3 、X 4 、X 5 、X 6 Independently selected from CR 5 or N; R 1 、R 2 independently selected from H, halogen, CN, OH, NH2, C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-10 membered heterocyclic group, the OH, NH2, C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-10 membered heterocyclic group is optionally replaced by R x replace; or R 1 、R 2 and the atoms they are connected to form C3-C 10 Cycloalkyl or 4-7 membered heterocyclic group, the C3-C 10 Cycloalkyl or 4-7 membered heterocyclic group is optionally replaced by R x replace; R 3 、R 4 、R 5 independently selected from H, halogen, CN, OH, NH2, -C(O)OR x 、-C(O)R x , C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C 10 Cycloalkyl, 4-7 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl, the OH, NH2, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C 10 Cycloalkyl, 4-7 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl is optionally replaced by R x replace; Ring A is selected from C6-C 10 Aryl or 5-10 membered heteroaryl, the C6-C 10 Aryl or 5-10 membered heteroaryl is optionally substituted by one or more R a replace; Ring B is selected from C6-C 10 Arylene, 5-10 membered heteroarylene, 4-10 membered heterocyclylene, C4-C 10 Cycloalkenylene or C3-C 10 Cycloalkylene, the C6-C 10 Arylene, 5-10 membered heteroarylene, 4-10 membered heterocyclylene, C4-C 10 Cycloalkenylene or C3-C 10 Cycloalkylene is optionally substituted with one or more R b replace; Ring C is selected from C3-C 10 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl, the C3-C 10 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl is optionally substituted by one or more R c replace; Every R a 、R b 、R c independently selected from halogen, CN, OH, NH2, -C(O)OR x , C1-C6 alkyl, -O-C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-10 membered heterocyclic group, the NH2, C1-C6 alkyl, -O-C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-10 membered heterocyclic group is optionally replaced by R x replace; or R b 、R c and the atoms they are connected to form C4-C 10 Cycloalkenyl or 4-10 membered heterocyclic group, the C4-C 10 Cycloalkenyl or 4-10 membered heterocyclic group is optionally replaced by R x replace; R x Selected from halogen, CN, OH, NH2, C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-8 membered heterocyclic group, the OH, NH2, C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-8 membered heterocyclic group is optionally replaced by R d replace; R d Selected from halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl or 4-8 membered heterocyclyl.

2. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R 1 、R 2 independently selected from H, halogen, C1-C6 alkyl or C3-C 10 Cycloalkyl, the C1-C6 alkyl or C3-C 10 The cycloalkyl group is optionally replaced by R x replace.

3. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1 or 2, characterized in that: R 3 、R 4 、R 5 independently selected from H, halogen, CN, OH, NH2, C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-7 membered heterocyclic group, the OH, NH2, C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-7 membered heterocyclic group is optionally replaced by R x replace.

4. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 3, characterized in that: R 3 、R 4 、R 5 Independently selected from H, C1-C6 alkyl or C3-C 10 Cycloalkyl, the C1-C6 alkyl or C3-C 10 The cycloalkyl group is optionally replaced by R x replace.

5. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, characterized in that: Ring A is selected from phenyl or 5-6 membered heteroaryl, wherein the phenyl or 5-6 membered heteroaryl is optionally substituted by one or more R a replace.

6. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, characterized in that: Ring B is selected from C6-C 10 Arylene or 5-10 membered heteroarylene, the C6-C 10 Arylene or 5-10 membered heteroarylene is optionally substituted by one or more R b replace.

7. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 6, characterized in that: Ring B is selected from C6-C 10 Arylene, the C6-C 10 The arylene group is optionally substituted by one or more R b replace.

8. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 7, characterized in that: Ring B is selected from phenylene, said phenylene being optionally substituted by one or more R b replace.

9. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 8, characterized in that: Ring C is selected from C6-C 10 Aryl or 5-10 membered heteroaryl, the C6-C 10 Aryl or 5-10 membered heteroaryl is optionally substituted by one or more R c replace.

10. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 9, characterized in that: Ring C is selected from 5-10 membered heteroaryl, said 5-10 membered heteroaryl being optionally substituted by one or more R c replace.

11. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 10, characterized in that: Every R a 、R b 、R c independently selected from halogen, C1-C6 alkyl, -O-C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-10 membered heterocyclic group, the C1-C6 alkyl, -O-C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-10 membered heterocyclic group is optionally replaced by R x replace.

12. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 11, characterized in that: Every R a 、R b 、R c independently selected from halogen, C1-C6 alkyl, -O-C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-6 membered heterocyclic group, wherein the C1-C6 alkyl, -O-C1-C6 alkyl, C3-C6 cycloalkyl or 4-6 membered heterocyclic group is optionally replaced by R x replace.

13. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 12, characterized in that: R b 、R c and the atoms to which they are attached together form a 4-10 membered heterocyclic group, the 4-10 membered heterocyclic group being optionally replaced by R x replace.

14. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 13, characterized in that: R x Selected from halogen, C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-8 membered heterocyclic group, the C1-C6 alkyl, C3-C 10 Cycloalkyl or 4-8 membered heterocyclic group is optionally replaced by R d replace; Or, R x is selected from halogen or C1-C6 alkyl, wherein the C1-C6 alkyl is optionally replaced by R d replace.

15. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 14, characterized in that X 1 、X 2 、X 4 、X 5 、X 6 Independently selected from N, X 3 Selected from CR 5 .

16. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 14, characterized in that: The compound represented by formula (I) or a pharmaceutically acceptable salt thereof is selected from the compound represented by formula (II) or a pharmaceutically acceptable salt thereof, Among them, ring A, ring B, ring C, R 1 、R 2 、R 3 、R 4 、X 1 、X 2 、X 3 and X 4 As defined in any one of claims 1 to 14.

17. 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, 18. A pharmaceutical composition comprising a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 17, and a pharmaceutically acceptable excipient.

19. Use of the compound of formula (I) or formula (II) or a pharmaceutically acceptable salt according to any one of claims 1 to 17, or the pharmaceutical composition according to claim 18, in the preparation of a medicament for preventing or treating a disease mediated by USP1.

20. A compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 17, or a pharmaceutical composition according to claim 18 for use in preventing or treating a disease mediated by USP1.

21. A method for preventing or treating a disease mediated by USP1, comprising administering a therapeutically effective dose of a compound of formula (I) or formula (II) according to any one of claims 1 to 17 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 18 to a mammal, preferably a human, in need of such treatment.

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