Thienopyrazole compounds, pharmaceutical compositions containing the same, and uses thereof

Abstract

The present invention belongs to the field of medicinal chemistry, relates to thiophenopyrazole compounds, pharmaceutical compositions containing the same and their uses. Specifically, the present invention provides a compound having the structure of formula I, which exhibits good anti-tumor activity.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical chemistry and relates to a thienopyrazole compound with UPS7 inhibitory activity, a pharmaceutical composition containing the same, and medical uses thereof. Background Art

[0002] The ubiquitin-proteasome system (UPS) is a basic physiological regulatory process in cells. Through a series of cascade reactions, proteins are ubiquitinated and then degraded by proteases. Abnormalities in the UPS are closely related to diseases such as tumors, neurodegenerative diseases, and viral infections. Currently, drugs are mainly developed for five types of targets in the UPS system: proteases, E1 activating enzymes, E2 conjugating enzymes, E3 ligases, and deubiquitinases (DUBs).

[0003] Deubiquitinating enzymes can specifically cut the isopeptide bond formed between the glycine residue at the carbon terminus of ubiquitin and the target protein, allowing ubiquitin to detach from the target protein, thereby protecting the target protein from degradation, relocation or activation, etc.

[0004] There are nearly 100 DUBs in the human body, among which ubiquitin-specific proteases (USPs) are the largest family of DUBs, including about 85 members, belonging to an isopeptidase in the cysteine ​​protease family (Wu and Kumar, Journal of Medicinal Chemistry, 2018, 61: 422-443). Currently, more than 40 USPs family members have been found to be associated with the occurrence and development of tumors.

[0005] USP7 is located in the cell nucleus and is a key deubiquitinase in the UPS. It can specifically cut the isopeptide bond formed between the carbon terminus of ubiquitin and the target protein, so that ubiquitin is separated from the target protein, thereby protecting the target protein from degradation, relocation or activation, etc. (Turnbull and Ioannidis, Nature, 2017, 550: 481-486).

[0006] USP7 is widely distributed in human tissues and plays an important role in neural development, cell cycle regulation, epigenetic regulation, DNA damage repair, and immune response. Studies have shown that USP7 is overexpressed in cancer cells such as hepatocellular carcinoma, multiple myeloma, colon cancer, lung cancer, prostate cancer, and bladder cancer, and this overexpression is directly related to tumor invasion and poor prognosis (Pozhidaeva and Bezsonova, DNA Repair, 2019, 76: 30-39).

[0007] USP7 has a wide variety of substrates, and most of them are proteins related to cell cycle regulation, immune response, apoptosis, and DNA damage repair, such as MDM2, p53, ERCC6, Foxp3, PTEN, and FOXO4, etc. (Chauhan and Tian, Cancer Cell, 2012, 22: 345 - 358). In some tumor cells, MDM2 is overexpressed. USP7 can protect MDM2 from ubiquitination. After MDM2 binds to the p53 protein, it promotes the ubiquitination and degradation of the p53 protein, thus promoting tumor growth.

[0008] USP7 can also play a role in promoting tumorigenesis by directly regulating the expression of tumor suppressor proteins (such as p53, PTEN, FOXO4, p114ARF, p16INK4) and tumor promoting proteins (such as N-MYC, REST), upregulating the expression of tumor-related factors (such as HIF-1), and regulating tumor-related signaling pathways (such as the SHH signaling pathway, Wnt / β-catenin signaling pathway, androgen receptor signaling pathway, DNA damage repair signaling pathway) (Zhou and Wang, Medicinal Chemistry, 2018, 14: 3 - 18).

[0009] In addition, USP7 also upregulates the activity of Treg cells by regulating upstream signaling molecules of Treg cells (such as the transcription factor FOXP3 and the epigenetic regulator Tip60) (Wang and Wu, PLoS One, 2017, 12: 1 - 23), and inhibits the activity of Teff cells (CD8+ T cells), thereby playing a role in tumor immune surveillance escape.

[0010] Developing inhibitors of USP7 is one of the hotspots in the field of tumor research. Currently, there is still no drug on the market targeting the USP7 target globally, and all the compounds under investigation are in the preclinical research stage.

[0011] Although companies such as Hybrigenics SA, Forma Therapeutics, Inc., Les Laboratoires Servier, and Almac Discovery Limited have conducted corresponding research on USP7 inhibitors and have published relevant patents, there is still an urgent need for new USP7 inhibitors in this field, especially USP7 inhibitors with high activity and other excellent properties. SUMMARY OF THE INVENTION

[0012] Problems to be Solved by the Invention

[0013] Through extensive research, the present invention has developed a thienopyrazole compound and its corresponding preparation method, which can significantly inhibit the activity of UPS7. As a UPS7 inhibitor, it can be used to prevent and / or treat diseases at least partially mediated by UPS7, especially tumor diseases (cancer).

[0014] Solutions for Solving the Problems

[0015] In a first aspect, the present invention provides a compound having a structure of Formula I or a pharmaceutically acceptable salt, ester, solvate, stereoisomer, tautomer, polymorph, isotope-labeled substance, metabolite, prodrug or mixture thereof,

[0016]

[0017] in,

[0018] R 0 Selected from hydrogen, halogen, cyano, C 1-6 Alkyl and C 1-6 Haloalkyl;

[0019] L 1 Not present or selected from C 1-6 Alkylene, C 1-6 Halogenated alkylene, C 3-8 Cycloalkylene and 3-8 membered heterocycloalkylene;

[0020] R 1 Selected from hydrogen, hydroxyl, halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 6-10 Aryl, 5-10 membered heteroaryl, -NR a R b , in,

[0021] R a and R b are each independently selected from hydrogen, C 1-6 Alkyl, C 3-8 Cycloalkyl, -C(O)-C 1-6 Alkyl, -C(O)-C 3-8 Cycloalkyl, -C(O)-C 2-6 Alkenyl, -S(O) q -C 1-6 Alkyl, -S(O) q -C 3-8 Cycloalkyl and -S(O) q -C 2-6 alkenyl; wherein the C 1-6 Alkyl, C3-8 The cycloalkyl and C 2-6 alkenyl are each independently optionally substituted by one or more of the following groups: hydrogen, -C 1-6 alkylene-C(O)OH and -C(O)O-C 1-6 alkyl; q is selected from 1 and 2;

[0022] B is selected from 3- to 8-membered heterocycloalkyl, 3- to 8-membered heterocycloalkenyl, and 5- to 10-membered heteroaryl;

[0023] C is selected from C 3-8 cycloalkyl, 3- to 8-membered heterocycloalkyl, C 4-6 cycloalkenyl, 3- to 8-membered heterocycloalkenyl, C 6-10 aryl, and 5- to 10-membered heteroaryl;

[0024] D is selected from C 3-8 cycloalkyl and 3- to 8-membered heterocycloalkyl;

[0025] If present, each R 9 and R 10 are each independently selected from hydrogen, halogen, cyano, oxo, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl, 3- to 8-membered heterocycloalkyl, carboxyl, -C(O)O-C 1-6 alkyl, and -C 1-6 alkylene-C(O)OH; wherein the C 1-6 alkyl, C 3-8 cycloalkyl, and 3- to 8-membered heterocycloalkyl are each independently optionally substituted by one or more of the following groups: hydrogen, hydroxy, and amino;

[0026] z 3 、z 4 and z 5 are each independently selected from 0, 1, 2, 3, 4, 5, and 6;

[0027] R 2 、R 3 、R 4 、R 5 and R 6 are each independently selected from hydrogen, hydroxy, halogen, cyano, carboxyl, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl, C 1-6 alkoxy, and C 1-6 haloalkoxy;

[0028] L 2 is selected from oxygen, sulfur, -N(R 11 )-, -C(O)-, -O-C 1-6Alkylene-, -S-C 1-6 Alkylene- and C 1-6 Alkylene; wherein,

[0029] R 11 is selected from hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl and -C(O)-C 1-6 alkyl;

[0030] A is selected from C 3-8 cycloalkyl, 3- to 8-membered heteroalkyl, C 6-10 aryl and 5- to 10-membered heteroaryl;

[0031] When present, each R 7 and R 8 is independently selected from hydrogen, hydroxy, halogen, cyano, -NR c R d 、C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 3-8 cycloalkyl, 3- to 8-membered heteroalkyl, -C 1-6 alkylene-OH, -C 1-6 alkylene-O-C 1-6 alkyl, -C(O)-N(C 1-6 alkyl)2, -C(O)-C 1-6 alkyl and -C(O)-C 1-6 haloalkyl, or R 7 and R 8 and the atoms to which they are attached together form C 3-8 cycloalkyl or 3- to 8-membered heteroalkyl; wherein,

[0032] R c and R d are independently selected from hydrogen, C 1-6 alkyl and -C(O)-C 1-6 alkyl;

[0033] z 1 and z 2 are independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10.

[0034] In a second aspect, the present invention provides specific compounds having the structure of formula I, which include:

[0035] (1) 3 - ((3 - (5 - chloro - 3 - methyl - 2 - (piperidin - 4 - yloxy)phenyl)-2 - methyl - 2H - thiophene[3,2 - c]pyrazol - 5 - yl)methyl)-6,6 - dimethyl - 3 - azabicyclo[3.1.0]hexane - 2,4 - dione;

[0036] (2) 3 - ((3 - (5 - chloro - 3 - methyl - 2 - (2 - methylpiperidin - 4 - yloxy)phenyl)-2 - methyl - 2H - thiophene[3,2 - c]pyrazol - 5 - yl)methyl)-6,6 - dimethyl - 3 - azabicyclo[3.1.0]hexane - 2,4 - dione;

[0037] (3) 3 - ((3 - (5 - chloro - 3 - methyl - 2 - (3 - methylpiperidin - 4 - yloxy)phenyl)-2 - methyl - 2H - thiophene[3,2 - c]pyrazol - 5 - yl)methyl)-6,6 - dimethyl - 3 - azabicyclo[3.1.0]hexane - 2,4 - dione;

[0038] (4) 3 - ((3 - (5 - chloro - 3 - methyl - 2 - (piperidin - 4 - ylamino)phenyl)-2 - methyl - 2H - thiophene[3,2 - c]pyrazol - 5 - yl)methyl)-6,6 - dimethyl - 3 - azabicyclo[3.1.0]hexane - 2,4 - dione;

[0039] (5) 3 - ((3 - (5 - chloro - 3 - methyl - 2 - (piperidine - 4 - carbonyl)phenyl)-2 - methyl - 2H - thiophene[3,2 - c]pyrazol - 5 - yl)methyl)-6,6 - dimethyl - 3 - azabicyclo[3.1.0]hexane - 2,4 - dione;

[0040] (6) 3 - ((3 - (5 - chloro - 3 - methyl - 2 - (piperidin - 4 - ylmethyl)phenyl)-2 - methyl - 2H - thiophene[3,2 - c]pyrazol - 5 - yl)methyl)-6,6 - dimethyl - 3 - azabicyclo[3.1.0]hexane - 2,4 - dione;

[0041] (7) 3 - ((3 - (5 - chloro - 2 - (2,6 - dimethylpiperidin - 4 - yloxy)-3 - methylphenyl)-2 - methyl - 2H - thiophene[3,2 - c]pyrazol - 5 - yl)methyl)-6,6 - dimethyl - 3 - azabicyclo[3.1.0]hexane - 2,4 - dione;

[0042] (8) 3 - ((3 - (5 - chloro - 3 - methyl - 2 - (1 - methylpiperidin - 4 - yloxy)phenyl)-2 - methyl - 2H - thiophene[3,2 - c]pyrazol - 5 - yl)methyl)-6,6 - dimethyl - 3 - azabicyclo[3.1.0]hexane - 2,4 - dione;

[0043] (9) 3 - ((3 - (5 - chloro - 3 - methyl - 2 - (6 - methylpiperidin - 3 - yloxy)phenyl) - 2 - methyl - 2H - thiopheno[3,2 - c]pyrazol - 5 - yl)methyl) - 6,6 - dimethyl - 3 - azabicyclo[3.1.0]hexane - 2,4 - dione;

[0044] (10) 3 - ((3 - (5 - chloro - 3 - methyl - 2 - (quinolin - 4 - yloxy)phenyl) - 2 - methyl - 2H - thiopheno[3,2 - c]pyrazol - 5 - yl)methyl) - 6,6 - dimethyl - 3 - azabicyclo[3.1.0]hexane - 2,4 - dione;

[0045] (11) 3 - ((3 - (2 - (1 - azabicyclo[3.2.1]octan - 5 - yloxy) - 5 - chloro - 3 - methylphenyl) - 2 - methyl - 2H - thiopheno[3,2 - c]pyrazol - 5 - yl)methyl) - 6,6 - dimethyl - 3 - azabicyclo[3.1.0]hexane - 2,4 - dione;

[0046] (12) 3 - ((3 - (2 - (3 - aminocyclopentyloxy) - 5 - chloro - 3 - methylphenyl) - 2 - methyl - 2H - thiopheno[3,2 - c]pyrazol - 5 - yl)methyl) - 6,6 - dimethyl - 3 - azabicyclo[3.1.0]hexane - 2,4 - dione;

[0047] (13) 3 - ((3 - (5 - chloro - 2 - (2,2 - dimethylpyrrolidin - 3 - yloxy) - 3 - methylphenyl) - 2 - methyl - 2H - thiopheno[3,2 - c]pyrazol - 5 - yl)methyl) - 6,6 - dimethyl - 3 - azabicyclo[3.1.0]hexane - 2,4 - dione;

[0048] (14) 3 - ((3 - (5 - chloro - 2 - (4 - methoxypyrrolidin - 3 - yloxy) - 3 - methylphenyl) - 2 - methyl - 2H - thiopheno[3,2 - c]pyrazol - 5 - yl)methyl) - 6,6 - dimethyl - 3 - azabicyclo[3.1.0]hexane - 2,4 - dione;

[0049] (15) 3 - ((3 - (5 - chloro - 2 - (1,4 - dimethylpyrrolidin - 3 - yloxy) - 3 - methylphenyl) - 2 - methyl - 2H - thiopheno[3,2 - c]pyrazol - 5 - yl)methyl) - 6,6 - dimethyl - 3 - azabicyclo[3.1.0]hexane - 2,4 - dione;

[0050] (16) 3 - ((3 - (2 - ((3 - aminocyclobutyl)methoxy) - 5 - chloro - 3 - methylphenyl) - 2 - methyl - 2H - thiopheno[3,2 - c]pyrazol - 5 - yl)methyl) - 6,6 - dimethyl - 3 - azabicyclo[3.1.0]hexane - 2,4 - dione; and

[0051] (17) 3 - ((3 - (5 - chloro - 3 - methyl - 2 - ((3 - methylazetidin - 3 - yl)methoxy)phenyl) - 2 - methyl - 2H - thiopheno[3,2 - c]pyrazol - 5 - yl)methyl) - 6,6 - dimethyl - 3 - azabicyclo[3.1.0]hexane - 2,4 - dione.

[0052] In a third aspect, the present invention provides a pharmaceutical composition comprising the compound having the structure of formula I above or a pharmaceutically acceptable salt, ester, solvate, stereoisomer, tautomer, polymorph, isotopically labeled compound, metabolite, prodrug thereof or a mixture thereof, and one or more pharmaceutically acceptable carriers.

[0053] In a fourth aspect, the present invention provides a medicament comprising:

[0054] a) the compound having the structure of formula I above or a pharmaceutically acceptable salt, ester, solvate, stereoisomer, tautomer, polymorph, isotopically labeled compound, metabolite, prodrug thereof or a mixture thereof, or the above - mentioned pharmaceutical composition;

[0055] b) optionally present packaging and / or instructions.

[0056] In a fifth aspect, the present invention provides the compound having the structure of formula I above or a pharmaceutically acceptable salt, ester, solvate, stereoisomer, tautomer, polymorph, isotopically labeled compound, metabolite, prodrug thereof or a mixture thereof, or the above - mentioned pharmaceutical composition, or the above - mentioned medicament, which is used as a UPS7 inhibitor for preventing and / or treating diseases or disorders (especially cancer) mediated at least in part by UPS7, or for inhibiting UPS7 activity in vitro or inhibiting tumor cell proliferation in vitro.

[0057] In a sixth aspect, the present invention provides the use of the compound having the structure of formula I above or a pharmaceutically acceptable salt, ester, solvate, stereoisomer, tautomer, polymorph, isotopically labeled compound, metabolite, prodrug thereof or a mixture thereof, or the above - mentioned pharmaceutical composition, or the above - mentioned medicament, as a UPS7 inhibitor.

[0058] In a seventh aspect, the present invention provides the use of the compound having the structure of formula I above or a pharmaceutically acceptable salt, ester, solvate, stereoisomer, tautomer, polymorph, isotopically labeled compound, metabolite, prodrug thereof or a mixture thereof, or the above - mentioned pharmaceutical composition, or the above - mentioned medicament, in the preparation of a medicament for preventing and / or treating diseases or disorders (especially cancer) mediated at least in part by UPS7.

[0059] In the eighth aspect, the present invention provides a method for preventing and / or treating a disease or condition (especially cancer) that is at least partially mediated by UPS7, comprising the following steps: administering a preventive and / or therapeutically effective amount of the above-mentioned compound having the structure of Formula I or its pharmaceutically acceptable salt, ester, solvate, stereoisomer, tautomer, polymorph, isotope-labeled substance, metabolite, prodrug or a mixture thereof, or the above-mentioned pharmaceutical composition, or the above-mentioned medicine to an individual in need thereof.

[0060] Effects of the Invention

[0061] The present invention provides a novel thienopyrazole compound, which can be used as an efficient UPS7 inhibitor and has anti-tumor activity. In addition, the synthesis method of the compound is mild and easy to operate. DETAILED DESCRIPTION

[0062] Before further describing the present invention, it should be understood that the present invention is not limited to the specific embodiments described herein. It should also be understood that, unless otherwise defined, the meaning or intention of all terms used herein are the same as those generally understood by those skilled in the art. Although the meaning or intention of the terms used herein are easily understood by those skilled in the art, in order to better explain the technical solution of the present invention, it is still described as follows.

[0063] [Definition of terms]

[0064] The terms "comprising," "including," "having," "involving," or any other variation thereof, are intended to cover a non-exclusive or open-ended inclusion. For example, a composition, method, or apparatus that comprises a list of elements is not necessarily limited to only the elements expressly listed but may also include other elements not expressly listed or inherent to the composition, method, or apparatus described above.

[0065] The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention that is substantially non-toxic to an organism. Pharmaceutically acceptable salts generally include (but are not limited to) salts formed by reacting a compound of the present invention with a pharmaceutically acceptable inorganic / organic acid or inorganic / organic base, such salts being also referred to as acid addition salts or base addition salts.

[0066] The term "pharmaceutically acceptable ester" refers to an ester that is substantially non-toxic to an organism and is hydrolyzed in vivo to form a compound of the present invention or a salt thereof. Pharmaceutically acceptable esters generally include (but are not limited to) esters formed between a compound of the present invention and a pharmaceutically acceptable carboxylic acid or sulfonic acid, which are also referred to as carboxylic acid esters or sulfonic acid esters.

[0067] The term "isomer" refers to compounds that have the same molecular weight due to having the same number and types of atoms, but differ in the spatial arrangement or configuration of the atoms.

[0068] The term "stereoisomer" (or "optical isomer") refers to stable isomers that have a perpendicular plane of asymmetry due to having at least one chiral element (including chiral centers, chiral axes, chiral planes, etc.), and thus can rotate plane-polarized light. Since the compounds of the present invention have asymmetric centers and other chemical structures that may lead to stereoisomerism, the present invention also includes these stereoisomers and their mixtures. Since the compounds of the present invention (or their pharmaceutically acceptable salts) contain asymmetric carbon atoms, they can exist in the form of a single stereoisomer, a racemate, a mixture of enantiomers and diastereomers. Generally, these compounds can be prepared in the form of a racemate. However, if desired, such compounds can be prepared or separated to obtain pure stereoisomers, i.e., single enantiomers or diastereomers, or mixtures enriched in a single stereoisomer (purity ≥98%, ≥95%, ≥93%, ≥90%, ≥88%, ≥85% or ≥80%). As described hereinafter, the single stereoisomers of the compounds are synthesized from optically active starting materials containing the desired chiral centers, or are prepared by separating or resolving a mixture of enantiomeric products, such as converting to a mixture of diastereomers and then separating or recrystallizing, chromatographing, using chiral resolving agents, or directly separating the enantiomers on a chiral chromatographic column. The starting compounds with specific stereochemistry can be either commercially available or prepared according to the methods described hereinafter and then resolved by methods well known in the art. The term "enantiomer" refers to a pair of stereoisomers that are non-superimposable mirror images of each other. The term "diastereomer" or "diastereoisomer" refers to optically active isomers that are not mirror images of each other. The term "racemic mixture" or "racemate" refers to a mixture containing equal amounts of single enantiomers (i.e., an equimolar mixture of the two R and S enantiomers). The term "non-racemic mixture" refers to a mixture containing unequal amounts of single enantiomers. Unless otherwise indicated, all stereoisomeric forms of the compounds of the present invention are within the scope of the present invention.

[0069] The term "tautomer" (or "tautomeric form") refers to structural isomers with different energies that can interconvert via a low energy barrier. If tautomerism is possible (e.g., in solution), a chemical equilibrium of tautomers can be achieved. For example, proton tautomers (or prototropic tautomers) include (but are not limited to) interconversions that occur via proton migration, such as keto-enol isomerization, imine-enamine isomerization, amide-imidol isomerization, etc. Unless otherwise indicated, all tautomeric forms of the compounds of the present invention are within the scope of the present invention.

[0070] The term "polymorph" (or "polymorphic form") refers to the solid crystalline forms of a compound or complex. Those skilled in the art can obtain polymorphs of a molecule by many known methods. These methods include (but are not limited to) melt recrystallization, melt cooling, solvent recrystallization, desolvation, rapid evaporation, rapid cooling, slow cooling, vapor diffusion, and sublimation. Additionally, well-known techniques can be used to detect, classify, and identify polymorphs, including (but not limited to) differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), X-ray powder diffraction (XRPD), single crystal X-ray diffraction (SCXRD), solid state nuclear magnetic resonance (NMR), infrared spectroscopy (IR), Raman spectroscopy, and scanning electron microscopy (SEM), etc.

[0071] The term "solvate" refers to a substance formed by the binding of a compound of the present invention (or its pharmaceutically acceptable salt) to at least one solvent molecule through non-covalent intermolecular forces. Common solvates include (but are not limited to) hydrates (including hemihydrates, monohydrates, dihydrates, trihydrates, etc.), ethanolates, acetonates, etc.

[0072] The term "isotope-labeled compound" refers to a derivative compound formed by replacing a specific atom in a compound of the present invention with its isotope atom. Unless otherwise indicated, the compounds of the present invention include various isotopes of H, C, N, O, F, P, S, Cl, such as 2 H(D), 3 H(T), 13 C, 14 C, 15 N, 17 O, 18 O, 18 F, 31 P, 32 P, 35 S, 36 S and 37 Cl. For example, 12 C can be replaced by 12 C, 13 C or 14 C; 1 H can be replaced by2 H(D, deuterium) or 3 H(T, tritium) substitution; 16 O can be replaced by 18 O, etc. The present invention includes isotopically labeled compounds obtained by replacing any atom in the structure with its isotope.

[0073] The term "metabolite" refers to a derivative compound formed after metabolism of the compound of the present invention. Further information on metabolism can be found in Goodman and Gilman's: The Pharmacological Basis of Therapeutics (9 th th ed.) [M], McGraw-Hill International Editions, 1996.

[0074] The term "prodrug" refers to a derivative compound that can directly or indirectly provide the compound of the present invention when administered to an individual. Particularly preferred derivative compounds or prodrugs are compounds that can improve the bioavailability of the compound of the present invention (e.g., more easily absorbed into the blood) when administered to an individual, or compounds that promote the delivery of the parent compound to the site of action (e.g., the lymphatic system). Unless otherwise indicated, all prodrug forms of the compounds of the present invention are within the scope of the present invention, and various prodrug forms are well known in the art.

[0075] The term "independently of each other" means that at least two groups (or ring systems) with the same or similar range of values present in the structure can have the same or different meanings in a specific situation. For example, if substituents X and Y are independently of each other hydrogen, halogen, hydroxyl, cyano, alkyl or aryl, then when substituent X is hydrogen, substituent Y can be either hydrogen or halogen, hydroxyl, cyano, alkyl or aryl; similarly, when substituent Y is hydrogen, substituent X can be either hydrogen or halogen, hydroxyl, cyano, alkyl or aryl.

[0076] The term "alkyl" refers to a straight-chain or branched-chain saturated aliphatic hydrocarbon group. For example, the term "C 1-6 alkyl" used in the present invention refers to an alkyl group having 1 to 6 carbon atoms (such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, etc.), which is optionally substituted by one or more (e.g., 1 to 3) substituents described in the present invention (when substituted by halogen, the group is "C 1-6 haloalkyl", e.g., -CF3, -C2F5, -CHF2, -CH2F, -CH2CF3, -CH2Cl, -CH2CH2CF3, etc.).

[0077] The term "alkylene" refers to a divalent straight-chain or branched-chain saturated aliphatic hydrocarbon group, where the two groups or segments it is attached to can be attached to the same carbon atom or different carbon atoms. For example, the term "C 1-6 alkylene" used in the present invention refers to an alkylene having 1 to 6 carbon atoms (such as methylene, 1,1-ethylene, 1,2-ethylene, etc.), which is optionally substituted by one or more (such as 1 to 3) substituents described in the present invention (when substituted by a halogen, the group is a "C 1-6 haloalkylene", such as -CF2-, -C2F4-, -CHF-, etc.).

[0078] The term "cycloalkyl" refers to a monocyclic or polycyclic (such as bicyclic) aliphatic hydrocarbon group (such as a monocyclic cycloalkyl, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, etc.; or a bicyclic cycloalkyl, including fused rings, bridged rings or spiro rings, such as bicyclo[1.1.1]pentyl, bicyclo[2.2.1]heptyl, etc.), which is optionally substituted by one or more (such as 1 to 3) substituents described in the present invention (such as an oxo group). For example, the term "C 3-8 cycloalkyl" used in the present invention refers to a cycloalkyl having 3 to 8 ring-forming carbon atoms (such as 2-methylcyclopropyl).

[0079] The term "cycloalkylene" refers to a divalent monocyclic or polycyclic (such as bicyclic) aliphatic hydrocarbon group, where the two groups or segments it is attached to can be attached to the same ring-forming carbon atom or different ring-forming carbon atoms, and it is optionally substituted by one or more (such as 1 to 3) substituents described in the present invention (such as an oxo group). For example, the term "C 3-8 cycloalkylene" used in the present invention refers to a cycloalkylene having 3 to 8 ring-forming carbon atoms (such as ).

[0080] The term "heterocycloalkyl" refers to a monocyclic or polycyclic (such as bicyclic) aliphatic group having one or more carbon atoms (e.g., 2, 3, 4, 5, 6, 7, 8, or 9) and one or more (e.g., 2, 3, or 4) heteroatoms or heteroatom groups each independently selected from O, S, N, NH, S(O), and S(O)2 in the ring, which is optionally substituted with one or more (e.g., 1 to 3) substituents described in the present invention (e.g., oxo group). If the valence requirements are met, the heterocycloalkyl can be linked to the parent molecular moiety through any one carbon atom or heteroatom in the ring. For example, the term "3-8 membered heterocycloalkyl" used in the present invention refers to a heterocycloalkyl having 3 to 8 ring atoms (including one or more heteroatoms or heteroatom groups) (e.g., oxiranyl, aziridinyl, azetidinyl, oxetanyl, tetrahydrofuryl, pyrrolidinyl, pyrrolidinonyl, imidazolidinyl, pyrazolidinyl, pyrrolidinyl, tetrahydropyranyl, etc.).

[0081] The term "heterocycloalkylidene" refers to a divalent monocyclic or polycyclic (such as bicyclic) aliphatic hydrocarbon group having one or more carbon atoms (e.g., 2, 3, 4, 5, 6, 7, 8, or 9) and one or more (e.g., 2, 3, or 4) heteroatoms or heteroatom groups each independently selected from O, S, N, NH, S(O), and S(O)2 in the ring, and the two groups or segments to which it is attached can be linked to the same ring-forming carbon atom or can be linked to a ring-forming carbon atom and a ring-forming heteroatom (e.g., nitrogen atom) respectively, which is optionally substituted with one or more (e.g., 1 to 3) substituents described in the present invention (e.g., oxo group). For example, the term "3-8 membered heterocycloalkylidene" used in the present invention refers to a heterocycloalkylidene having 3 to 8 ring atoms (including one or more heteroatoms or heteroatom groups) (e.g., ).

[0082] The term "alkoxy" refers to an "alkyl" or "cycloalkyl" as defined above that is linked to the parent molecular moiety through an oxygen atom (e.g., C 1-6 alkoxy, C 3-8 cycloalkoxy, etc., including methoxy, ethoxy, n-propoxy, isopropoxy, cyclopropoxy, n-butoxy, isobutoxy, tert-butoxy, sec-butoxy, cyclobutoxy, n-pentyloxy, isopentyloxy, n-hexyloxy or its isomers). For example, the term "C 1-6"Alkoxy" refers to an alkoxy group having 1 to 6 carbon atoms (such as methoxy, ethoxy, tert-butoxy, etc.), which is optionally substituted by one or more (such as 1 to 3) substituents described in the present invention (when substituted by a halogen, the group is "C 1-6 haloalkoxy", such as -OCF3, -OC2F5, etc.).

[0083] The term "aryl" refers to a monocyclic or fused polycyclic aromatic hydrocarbon group having a conjugated π electron system, which is optionally substituted by one or more substituents described in the present invention (such as being substituted by an oxo group, C 1-6 alkyl (methyl, ethyl) or halogen (fluorine, chlorine, bromine)). For example, the term "C 6-10 aryl" used in the present invention refers to an aryl group having 6 to 10 carbon atoms (such as phenyl, naphthyl, etc.).

[0084] The term "heteroaryl" refers to a monocyclic or fused polycyclic aromatic group having a conjugated π electron system, and one or more (such as 1, 2, 3, 4, 5, 6, 9 or 10) carbon atoms and one or more (such as 2, 3 or 4) heteroatoms or heteroatom groups each independently selected from O, S, N and NH are present in its ring, which is optionally substituted by one or more substituents described in the present invention (such as being substituted by an oxo group, C 1-6 alkyl (methyl, ethyl) or halogen (fluorine, chlorine, bromine)). If the valence bond requirements are met, the heteroaryl can be attached to the parent molecular moiety through any ring atom. For example, the term "5-10 membered heteroaryl" used in the present invention refers to a heteroaryl having 5 to 10 ring atoms (such as thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl or its benzo derivatives, etc.).

[0085] The term "alkenyl" refers to a straight-chain or branched-chain aliphatic hydrocarbon group containing one or more carbon-carbon double bonds, usually containing about 2 to 15 carbon atoms. For example, the term "C 2-6"Alkenyl" refers to an alkenyl group having 2 to 6 carbon atoms (such as vinyl, propenyl, n-butenyl, 3-methylbut-2-enyl, n-pentenyl, n-octenyl, n-decenyl, etc.), which is optionally substituted by one or more (such as 1 to 3) substituents (such as halogen, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, cyano, hydroxy, alkoxy, aryloxy, alkoxyalkyl, alkylthio, amino, alkylacyloxy, arylacyloxy, cycloalkylacyloxy, carboxyl, alkoxycarbonyl, etc.).

[0086] The term "cycloalkenyl" refers to a monocyclic or polycyclic (such as bicyclic) aliphatic hydrocarbon group having one or more carbon-carbon double bonds (such as monocyclic cycloalkenyl, such as cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, etc.). For example, the term "C 4-6 cycloalkenyl" used in the present invention refers to a cycloalkenyl group having 4 to 6 ring-forming carbon atoms, which is optionally substituted by one or more (such as 1 to 3) substituents described in the present invention (such as oxo group).

[0087] The term "heterocycloalkenyl" refers to a monocyclic or polycyclic (such as bicyclic) aliphatic group having one or more carbon atoms (such as 2, 3, 4, 5, 6, 7, 8 or 9) and one or more (such as 2, 3 or 4) heteroatoms or heteroatom groups each independently selected from O, S, N, NH, S(O) and S(O)2 in the ring, and having one or more carbon-carbon double bonds, which is optionally substituted by one or more (such as 1 to 3) substituents described in the present invention (such as oxo group). If the valence bond requirements are met, the heterocycloalkenyl can be connected to the parent molecular moiety through any one carbon atom or heteroatom in the ring. For example, the term "3-8 membered heterocycloalkenyl" used in the present invention refers to a heterocycloalkenyl group having 3 to 8 ring atoms (including one or more heteroatoms or heteroatom groups).

[0088] The term "amino" refers to the "-NH2" group, which can be either unsubstituted or substituted by one or more substituents described in the present invention (such as -NH2, -NHCH3, -N(CH3)2, -NH(C 1-3 alkyl), -N(C 1-3 alkyl)2, etc.). Unless otherwise specified, the term "amino" used in the present invention can also be protected by an amino protecting group. Suitable amino protecting groups include acetyl (Ac), tert-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz), etc.

[0089] The term "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br) and iodine (I).

[0090] The term "cyano" refers to the "-CN" group.

[0091] The term "hydroxy" refers to the "-OH" group.

[0092] The term "carboxyl group" refers to the "-C(O)OH" group.

[0093] The term "oxo group" refers to the "=O" group. The oxo group usually appears on a carbon atom that is also connected to two other atoms, atomic groups (groups), or structural fragments, and together they form a carbonyl group.

[0094] The term "carbonyl group" refers to the "-C(=O)-" or "-C(O)-" group.

[0095] The term "substituted" means that one or more (e.g., 1, 2, 3, or 4) atoms (e.g., hydrogen atoms) or atomic groups (e.g., trifluoromethanesulfonate group) in the specified group are replaced by other atoms or atomic groups, provided that the specified group satisfies the valence requirements in the current situation and forms a stable compound after substitution. Combinations of substituents and / or variables are only permitted if the combination can form a stable compound. If a substituent is described as "optionally substituted by...", then the substituent can be unsubstituted or substituted. If the first substituent is described as optionally substituted by one or more of a list of second substituents, then one or more hydrogen atoms in the first substituent can be individually or independently replaced by one or more of the list of second substituents, or not replaced.

[0096] When the bond of a substituent is shown passing through a bond connecting two atoms in a ring, then such a substituent can be attached to any ring-forming atom in the ring.

[0097] The term "one or more" means 1 or more than 1 under reasonable conditions, such as 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0098] [General formula compound]

[0099] The present invention provides a compound of formula I or a pharmaceutically acceptable salt, ester, solvate (such as a hydrate), stereoisomer, tautomer, polymorph, isotope-labeled compound, metabolite, prodrug, or a mixture thereof.

[0100]

[0101] wherein,

[0102] R 0 is selected from hydrogen, halogen, cyano, C 1-6 alkyl, and C 1-6 haloalkyl;

[0103] L 1 is absent or is selected from C 1-6 alkylene, C 1-6 haloalkylene, C3-8 Subcycloalkyl and 3- to 8-membered heterocycloalkyl;

[0104] R 1 Selected from hydrogen, hydroxy, halogen, cyano, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 6-10 aryl, 5- to 10-membered heteroaryl, -NR a R b , wherein,

[0105] R a and R b are each independently selected from hydrogen, C 1-6 alkyl, C 3-8 cycloalkyl, -C(O)-C 1-6 alkyl, -C(O)-C 3-8 cycloalkyl, -C(O)-C 2-6 alkenyl, -S(O) q -C 1-6 alkyl, -S(O) q -C 3-8 cycloalkyl and -S(O) q -C 2-6 alkenyl; wherein, the C 1-6 alkyl, C 3-8 cycloalkyl and C 2-6 alkenyl are each independently optionally substituted with one or more of the following groups: hydrogen, -C 1-6 alkylene-C(O)OH and -C(O)O-C 1-6 alkyl; q is selected from 1 and 2;

[0106] B is selected from 3- to 8-membered heterocycloalkyl, 3- to 8-membered heterocycloalkenyl and 5- to 10-membered heteroaryl;

[0107] C is selected from C 3-8 cycloalkyl, 3- to 8-membered heterocycloalkyl, C 4-6 cycloalkenyl, 3- to 8-membered heterocycloalkenyl, C 6-10 aryl and 5- to 10-membered heteroaryl;

[0108] D is selected from C 3-8 cycloalkyl and 3- to 8-membered heterocycloalkyl;

[0109] If present, each R 9 and R 10 are each independently selected from hydrogen, halogen, cyano, oxo, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl, 3- to 8-membered heterocycloalkyl, carboxyl, -C(O)O-C1-6 alkyl and -C 1-6 alkylene -C(O)OH; wherein, said C 1-6 alkyl, C 3-8 cycloalkyl and 3 - 8 membered heteroalkyl are each independently optionally substituted with one or more of the following groups: hydrogen, hydroxy, and amino;

[0110] z 3 、z 4 and z 5 are each independently selected from 0, 1, 2, 3, 4, 5, and 6;

[0111] R 2 、R 3 、R 4 、R 5 and R 6 are each independently selected from hydrogen, hydroxy, halogen, cyano, carboxy, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl, C 1-6 alkoxy and C 1-6 haloalkoxy;

[0112] L 2 is selected from oxygen, sulfur, -N(R 11 )-, -C(O)-, -O-C 1-6 alkylene -, -S-C 1-6 alkylene - and C 1-6 alkylene; wherein,

[0113] R 11 is selected from hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl and -C(O)-C 1-6 alkyl;

[0114] A is selected from C 3-8 cycloalkyl, 3 - 8 membered heteroalkyl, C 6-10 aryl and 5 - 10 membered heteroaryl;

[0115] If present, each R 7 and R 8 are each independently selected from hydrogen, hydroxy, halogen, cyano, -NR c R d 、C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 3-8 cycloalkyl, 3 - 8 membered heteroalkyl, -C 1-6 alkylene -OH, -C 1-6 alkylene -O-C1-6 alkyl, -C(O)-N(C 1-6 alkyl)2, -C(O)-C 1-6 alkyl and -C(O)-C 1-6 haloalkyl, or R 7 and R 8 and the atoms to which they are attached together form a C 3-8 cycloalkyl or a 3- to 8-membered heterocycloalkyl; wherein,

[0116] R c and R d are each independently selected from hydrogen, C 1-6 alkyl and -C(O)-C 1-6 alkyl;

[0117] z 1 and z 2 are each independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10.

[0118] In some embodiments of the present invention, in the above formula I compound, R 0 is selected from hydrogen, halogen, C 1-4 alkyl, and C 1-4 haloalkyl.

[0119] In some embodiments of the present invention, in the above formula I compound, L 1 is absent or is selected from C 1-6 alkylene and C 1-6 haloalkylene; preferably, L 1 is absent or is selected from C 1-4 alkylene and C 1-4 haloalkylene.

[0120] In some embodiments of the present invention, in the above formula I compound, R 1 is selected from wherein,

[0121] B is selected from a 3- to 8-membered heterocycloalkyl and a 3- to 8-membered heterocycloalkenyl;

[0122] C is selected from C 3-8 cycloalkyl;

[0123] D is selected from C 3-8 cycloalkyl;

[0124] If present, each R 9 and R 10 is each independently selected from hydrogen, halogen, cyano, oxo, C 1-6 alkyl, C 1-6 haloalkyl, and carboxyl; wherein, the C 1-6The alkyl group is optionally substituted by one or more of the following groups: hydrogen, hydroxyl, and amino;

[0125] z 3 , z 4 and z 5 are each independently selected from 0, 1, and 2.

[0126] In some embodiments of the present invention, in the above formula I compound, R 2 , R 3 , R 4 , R 5 and R 6 are each independently selected from hydrogen, halogen, cyano, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, and C 1-6 haloalkoxy.

[0127] In some embodiments of the present invention, in the above formula I compound, L 2 is selected from oxygen, sulfur, -N(R 11 ), -C(O)-, -O-C 1-6 alkylene-, and C 1-6 alkylene; wherein,

[0128] R 11 is selected from hydrogen, C 1-6 alkyl, and C 1-6 haloalkyl.

[0129] In some embodiments of the present invention, in the above formula I compound, A is selected from C 3-8 cycloalkyl and 3-8 membered heteroalkyl.

[0130] In some embodiments of the present invention, in the above formula I compound, if present, each R 7 and R 8 are each independently selected from hydrogen, hydroxyl, halogen, cyano, -NR c R d , C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 3-8 cycloalkyl, and 3-8 membered heteroalkyl, or R 7 and R 8 and the atoms to which they are attached together form C 3-8 cycloalkyl or 3-8 membered heteroalkyl; preferably, if present, each R 7 and R 8 are each independently selected from hydrogen, hydroxyl, halogen, cyano, -NR c R d , C1-6 alkyl, C 1-6 alkoxy, and C 1-6 haloalkyl, or R 7 and R 8 together with the atoms to which they are attached form a C 3-8 cycloalkyl or 3- to 8-membered heterocycloalkyl; wherein,

[0131] R c and R d each independently selected from hydrogen, C 1-6 alkyl, and -C(O)-C 1-6 alkyl;

[0132] z 1 and z 2 each independently selected from 0, 1, and 2.

[0133] In some embodiments of the present invention, in the compound of formula I above,

[0134] R 0 is hydrogen;

[0135] L 1 is C 1-6 alkylene;

[0136] R 1 is selected from wherein,

[0137] B is selected from 3- to 8-membered heterocycloalkyl and 3- to 8-membered heterocycloalkenyl;

[0138] C is C 3-6 cycloalkyl;

[0139] If present, each R 9 is independently selected from oxo and C 1-6 haloalkyl;

[0140] If present, each R 10 is independently selected from hydrogen and C 1-6 alkyl;

[0141] R 2 , R 3 , R 4 , R 5 and R 6 are each independently selected from hydrogen, halogen, and C 1-6 alkyl;

[0142] L 2 is selected from -O-, -N(R 11 )-, -C(O)-, -O-C 1-6 alkylene-, and C 1-6 alkylene; wherein,

[0143] R 11 selected from hydrogen and C 1-6 alkyl;

[0144] A is selected from C 3-8 cycloalkyl and 3- to 8-membered heterocycloalkyl;

[0145] When present, each R 7 and R 8 are each independently selected from hydrogen, amino, C 1-6 alkyl, and C 1-6 alkoxy, or R 7 and R 8 together with the atoms to which they are attached form a C 3-8 cycloalkyl;

[0146] z 1 、z 2 、z 3 and z 4 are each independently selected from 0, 1, and 2.

[0147] In some preferred embodiments of the present invention, in the above compound of formula I,

[0148] R 0 is hydrogen;

[0149] L 1 is methylene;

[0150] R 1 is wherein,

[0151] B is 5-membered heterocycloalkyl;

[0152] C is cyclopropyl;

[0153] R 9 is oxo;

[0154] R 10 is C 1-6 alkyl;

[0155] z 3 and z 4 are 2;

[0156] R 2 、R 3 、R 4 、R 5 and R 6 are each independently selected from hydrogen, halogen, and C 1-6 alkyl;

[0157] L 2 is selected from -O-, -N(R 11)-, -C(O)-, -O-C 1-6 alkylene- and C 1-6 alkylene; wherein,

[0158] R 11 is selected from hydrogen and C 1-6 alkyl;

[0159] A is selected from C 4-5 cycloalkyl and 4- to 8-membered heterocycloalkyl;

[0160] When present, each R 7 and R 8 is independently selected from hydrogen, amino, C 1-6 alkyl and C 1-6 alkoxy;

[0161] z 1 and z 2 are independently selected from 0 and 1.

[0162] In some more preferred embodiments of the present invention, in the compound of formula I above,

[0163] R 0 is hydrogen;

[0164] L 1 is methylene;

[0165] R 1 is

[0166] R 2 and R 4 are hydrogen;

[0167] R 3 is chlorine;

[0168] R 5 and R 6 are methyl;

[0169] L 2 is selected from -O-, -NH-, -C(O)-, -O-CH2- and -CH2-;

[0170] A is selected from

[0171] In some preferred embodiments of the present invention, the compound of formula I above has the structure shown in formula II, wherein

[0172]

[0173] R 2 、R 3 、R 4 、R5 , R 7 , R 8 , L 2 , A, z 1 and z 2 as defined above.

[0174] In some other preferred embodiments of the present invention, the above compound of formula I has the structure shown in formula III, wherein

[0175]

[0176] R 0 , R 1 , R 6 , R 7 , R 8 , L 1 , L 2 , A, z 1 and z 2 as defined above.

[0177] In some other preferred embodiments of the present invention, the above compound of formula I has the structure shown in formula IV, wherein

[0178]

[0179] R 7 , R 8 , L 2 , A, z 1 and z 2 as defined above.

[0180] In some more preferred embodiments of the present invention, the above compound of formula I has the structure shown in any one of formulae IV-1 to IV-5, wherein

[0181]

[0182] R 7 , R 8 , A, z 1 and z 2 as defined above.

[0183] In addition, the present invention also provides the following compounds or their pharmaceutically acceptable salts, esters, solvates, stereoisomers, tautomers, polymorphs, isotope-labeled compounds, metabolites, prodrugs or mixtures thereof, the structures and names of which are shown in the following table:

[0184]

[0185]

[0186]

[0187] [Preparation Method]

[0188] The present invention provides a preparation method of the above-mentioned compound of formula I.

[0189] The preparation method comprises the following steps:

[0190]

[0191] Step 1: React compound a with compound b to obtain compound c;

[0192] Step 2: Subject compound c to a substitution reaction to obtain compound d;

[0193] Step 3: React compound e with compound f to obtain compound g;

[0194] Step 4: React compound g with compound d to obtain compound h;

[0195] Step 5: Subject compound h to a reaction (such as a reduction reaction) to obtain compound i;

[0196] Step 6: Subject compound i to a substitution reaction to obtain compound j;

[0197] Step 7: React compound j with compound k to obtain the compound of formula I;

[0198] wherein, R 0 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , z 1 , z 2 , L 1 , L 2 and the definition of ring A are as described in formula I; X 1 , X 2 , X 3 and X 4 each independently selected from halogen and trifluoromethanesulfonyloxy, preferably X 1 , X 2 , X 3 and X 4 each independently selected from halogen; M is selected from boronic acid group, C 1-6 alkyl-substituted boronic acid group, a five-membered oxaborolane substituted with one or more methyl groups and C 1-6 alkyl-substituted tin group, preferably M is tri-n-butyltin.

[0199] In some embodiments of the present invention, the reaction in Step 1 is carried out in an organic solvent, and the organic solvent is selected from THF, DCM, and DMF, with THF being preferred.

[0200] In some embodiments of the present invention, the reaction in Step 1 is carried out in the presence of PPh3.

[0201] In some embodiments of the present invention, the reaction in Step 1 is carried out in the presence of an azo compound, and the azo compound is selected from DIAD, DEAD, and ADDP, with DIAD being preferred.

[0202] In some embodiments of the present invention, the reaction in Step 1 is carried out under the conditions of 0 °C to 50 °C, preferably under the conditions of 0 °C to 25 °C.

[0203] In some embodiments of the present invention, the substitution reaction in Step 2 is carried out in an organic solvent, and the organic solvent is selected from Tol, THF, and Et2O, with Tol being preferred.

[0204] In some embodiments of the present invention, the substitution reaction in Step 2 is carried out in the presence of a base, and the base is selected from n-BuLi, t-BuLi, LiHMDS, and NaHMDS, with n-BuLi being preferred.

[0205] In some embodiments of the present invention, the substitution reaction in Step 2 is carried out in the presence of a metal reagent, and the metal reagent is selected from tributyltin chloride, hexabutylditin, and tributylmethoxytin, with tributyltin chloride being preferred.

[0206] In some embodiments of the present invention, the reaction in Step 2 is carried out under the conditions of -78 °C to 50 °C, preferably under the conditions of -40 °C to 25 °C.

[0207] In some embodiments of the present invention, the reaction in Step 3 is carried out in an organic solvent, and the organic solvent is selected from alcohols (such as methanol, ethanol, isopropanol, etc.), EA, THF, ACN, DMF, DMSO, and Tol, with methanol being preferred.

[0208] In some embodiments of the present invention, the reaction in Step 3 is carried out in the presence of a base, and the base is selected from NaH, t-BuOK, K2CO3, Cs2CO3, MeONa, n-BuLi, DIPEA, and Et3N, with MeONa being preferred.

[0209] In some embodiments of the present invention, the reaction in Step 3 is carried out under the conditions of -20 °C to 100 °C, preferably under the conditions of 0 °C to 25 °C.

[0210] In some embodiments of the present invention, the reaction in Step 4 is carried out in an organic solvent, and the organic solvent is selected from Diox, Tol, ACN, DMF, and THF, preferably Diox.

[0211] In some embodiments of the present invention, the reaction in Step 4 is carried out under the catalysis of a catalyst, and the catalyst is selected from Pd(PPh3)4, Pd(OAc)2, Pd2(dba)3, Pd(PPh3)2Cl2, Pd(PPh3)2Cl2·CH2Cl2, and Pd(dppf)Cl2, preferably Pd(PPh3)4.

[0212] In some embodiments of the present invention, the reaction in Step 4 is carried out in the presence of a copper reagent, and the copper reagent is selected from CuI, CuCl, and CuBr, preferably CuI.

[0213] In some embodiments of the present invention, the reaction in Step 4 is carried out under the condition of -20°C to 200°C, preferably under the condition of 80°C to 150°C.

[0214] In some embodiments of the present invention, the reaction in Step 5 is a reduction reaction.

[0215] In some preferred embodiments of the present invention, the reduction reaction in Step 5 is carried out in an organic solvent, and the organic solvent is selected from THF, DMF, DCM, Tol, Diox, and ACN, preferably THF.

[0216] In some preferred embodiments of the present invention, the reduction reaction in Step 5 is carried out in the presence of a reducing agent, and the reducing agent is selected from LiAlH4, DIBAL, and NaBH4, preferably LiAlH4.

[0217] In some preferred embodiments of the present invention, the reduction reaction in Step 5 is carried out under the condition of -20°C to 50°C, preferably under the condition of 0°C to 25°C.

[0218] In some embodiments of the present invention, the substitution reaction in Step 6 is carried out in an organic solvent, and the organic solvent is selected from DCM, DMF, THF, Et2O, and Tol, preferably DCM.

[0219] In some embodiments of the present invention, the substitution reaction in Step 6 is carried out in the presence of a halogenating reagent, and the halogenating reagent is selected from halogenated alkanes, trihalophosphines, trihaloxophosphines, hydrohalic acids, and DCSO, preferably DCSO.

[0220] In some embodiments of the present invention, the substitution reaction in Step 6 is carried out under the condition of -20°C to 50°C, preferably under the condition of 0°C to 25°C.

[0221] In some embodiments of the present invention, the reaction in step seven is carried out in an organic solvent, and the organic solvent is selected from DMF, DCM, THF, and Tol, preferably DMF.

[0222] In some embodiments of the present invention, the reaction in step seven is carried out in the presence of a base, and the base is selected from NaH, t-BuOK, Cs2CO3, and K2CO3, preferably NaH.

[0223] In some embodiments of the present invention, the reaction in step seven is carried out under the conditions of -20°C to 50°C, preferably under the conditions of 0°C to 25°C.

[0224] [Pharmaceutical composition]

[0225] The term "pharmaceutical composition" refers to a composition that can be used as a drug, which contains a pharmaceutically active ingredient (API) (or therapeutic agent) and optionally one or more pharmaceutically acceptable carriers, with the aim of facilitating the administration to an organism, promoting the absorption of the active ingredient, and thus exerting biological activity. The term "pharmaceutically acceptable carrier" refers to an excipient administered together with a therapeutic agent, and it is suitable for contacting the tissues of humans and / or other animals within the scope of reasonable medical judgment without excessive toxicity, irritation, allergic reaction, or other problems or complications corresponding to a reasonable benefit / risk ratio. Pharmaceutically acceptable carriers that can be used in the present invention include (but are not limited to): a) diluents; b) lubricants; c) binders; d) disintegrants; e) absorbents, colorants, flavoring agents, and sweetening agents; f) emulsifiers or dispersants; and / or g) reagents that enhance the absorption of the compound.

[0226] The present invention provides a pharmaceutical composition, which contains the above-mentioned compound of formula I or its pharmaceutically acceptable salt, ester, solvate (such as hydrate), stereoisomer, tautomer, polymorph, isotope-labeled compound, metabolite, prodrug, or a mixture thereof.

[0227] In some embodiments, the above-mentioned pharmaceutical composition further contains one or more pharmaceutically acceptable carriers.

[0228] The above-mentioned pharmaceutical composition can act systemically and / or locally. For this purpose, they can be administered by suitable routes, such as parenterally, topically, intravenously, orally, subcutaneously, intra-arterially, intradermally, transdermally, rectally, intracranially, intraperitoneally, intranasally, intramuscularly, by inhalation, or in all ways well-known to those skilled in the pharmaceutical field. The above-mentioned pharmaceutical composition can be administered in combination with at least one other therapeutic agent having a therapeutic effect on a disease or disorder.

[0229] The above-mentioned administration routes can be achieved through suitable dosage forms. The dosage forms that can be used in the present invention include (but are not limited to): tablets, capsules, lozenges, troches, powders, sprays, creams, ointments, suppositories, gels, pastes, lotions, ointments, aqueous suspensions, injectable solutions, elixirs, syrups, etc.

[0230] The above-mentioned pharmaceutical composition may contain 0.01 mg to 1000 mg of at least one of the above-mentioned compounds of formula I or its pharmaceutically acceptable salts, esters, solvates (such as hydrates), stereoisomers, tautomers, polymorphs, isotopically labeled compounds, metabolites, prodrugs or mixtures thereof.

[0231] The present invention also provides a method for preparing the above-mentioned pharmaceutical composition or its corresponding preparation form, which comprises combining at least one of the above-mentioned compounds of formula I or its pharmaceutically acceptable salts, esters, solvates (such as hydrates), stereoisomers, tautomers, polymorphs, isotopically labeled compounds, metabolites, prodrugs or mixtures thereof with one or more pharmaceutically acceptable carriers.

[0232] [Drug product]

[0233] The term "drug product" refers to a combination product comprising a therapeutic agent, optionally other therapeutic agents, and optionally packaging and / or instructions.

[0234] The present invention provides a drug product, which comprises:

[0235] a) The above-mentioned compound of formula I or its pharmaceutically acceptable salts, esters, solvates (such as hydrates), stereoisomers, tautomers, polymorphs, isotopically labeled compounds, metabolites, prodrugs or mixtures thereof, or the above-mentioned pharmaceutical composition;

[0236] b) Optionally present packaging and / or instructions.

[0237] The above-mentioned drug product may contain 0.01 mg to 1000 mg of at least one of the above-mentioned compounds of formula I or its pharmaceutically acceptable salts, esters, solvates (such as hydrates), stereoisomers, tautomers, polymorphs, isotopically labeled compounds, metabolites, prodrugs or mixtures thereof.

[0238] The present invention also provides a method for preparing the above-mentioned drug product, which comprises combining the above-mentioned compound of formula I or its pharmaceutically acceptable salts, esters, solvates (such as hydrates), stereoisomers, tautomers, polymorphs, isotopically labeled compounds, metabolites, prodrugs or mixtures thereof, or the above-mentioned pharmaceutical composition, with optionally present packaging and / or instructions.

[0239] [Medical use]

[0240] The compounds of the present invention can exhibit strong inhibitory effects on UPS7 and can be used as UPS7 inhibitors. Therefore, the present invention provides the use of the above-mentioned compound of formula I or its pharmaceutically acceptable salts, esters, solvates (such as hydrates), stereoisomers, tautomers, polymorphs, isotope-labeled compounds, metabolites, prodrugs or mixtures thereof, or the above-mentioned pharmaceutical compositions, or the above-mentioned medicaments, as UPS7 inhibitors.

[0241] In addition, the present invention also provides the use of the above-mentioned compound of formula I or its pharmaceutically acceptable salts, esters, solvates (such as hydrates), stereoisomers, tautomers, polymorphs, isotope-labeled compounds, metabolites, prodrugs or mixtures thereof, or the above-mentioned pharmaceutical compositions, or the above-mentioned medicaments, in the preparation of a medicament for preventing and / or treating a disease or disorder (especially cancer) that is at least partially mediated by UPS7.

[0242] The term "disease or disorder that is at least partially mediated by UPS7" refers to a disease whose pathogenesis involves at least some factors related to UPS7. These diseases include (but are not limited to) cancer, neurodegenerative diseases (such as Alzheimer's disease, Parkinson's disease, etc.), diabetes, bone and joint diseases, joint inflammatory disorders, osteoporosis, immune disorders, cardiovascular diseases, ischemic diseases, viral infections and diseases, bacterial infections and diseases, etc.

[0243] [Therapeutic method]

[0244] The present invention provides a method for preventing and / or treating a disease (especially cancer) that is at least partially mediated by UPS7, which comprises the following step: administering a prophylactically and / or therapeutically effective amount of the above-mentioned compound of formula I or its pharmaceutically acceptable salts, esters, solvates (such as hydrates), stereoisomers, tautomers, polymorphs, isotope-labeled compounds, metabolites, prodrugs or mixtures thereof, or the above-mentioned pharmaceutical compositions, or the above-mentioned medicaments, to an individual in need thereof.

[0245] The term "effective amount" refers to a dose that is capable of inducing a biological or medical response in a cell, tissue, organ, or organism (e.g., an individual) and that is sufficient to achieve the desired prophylactic and / or therapeutic effect. The dosing regimen can be adjusted to provide an optimal response. For example, it can be administered as a single dose, in divided doses over time, or the dose can be proportionally decreased or increased depending on the circumstances. It is understood that for any particular individual, the specific dosing regimen should be adjusted according to need and the professional judgment of the administering person. Additionally, a distinction is made between prophylactic and therapeutic applications. In prophylactic applications, relatively low doses are typically administered over a relatively long interval for an extended period. In therapeutic applications, relatively high doses are typically administered at relatively short intervals until the progression of the disease is retarded or stopped, preferably until the individual shows partial or complete improvement of the symptoms.

[0246] In the present invention, suitable in vitro or in vivo assays are conducted to determine the effectiveness of the compound, pharmaceutical composition, and / or medicament and whether the administration is suitable for treating a disease or disorder afflicting an individual. Examples of these assays will be described in the non-limiting examples below. Generally, the effective amount of a compound sufficient to achieve a prophylactic and / or therapeutic effect is from about 0.001 mg / kg body weight / day to about 10,000 mg / kg body weight / day. In suitable cases, the effective amount is from about 0.01 mg / kg body weight / day to about 1,000 mg / kg body weight / day. In more general cases, the effective amount is about 0.01 - 1,000 mg / kg body weight per day, every two days, or every three days, preferably about 0.1 - 500 mg / kg body weight. Exemplary treatment regimens are administration once every two days, once a week, or once a month.

[0247] The term "treatment" refers to alleviating or eliminating the disease or disorder being targeted. If a subject has received a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable form thereof or a pharmaceutical composition of the present invention and at least one of the subject's indicators and symptoms shows an observable and / or detectable remission and / or improvement, it indicates that the subject has been successfully "treated". It is understood that treatment includes not only complete treatment but also treatment that does not achieve complete treatment but achieves some biologically or medically relevant result. Specifically, "treatment" means that the compound of the present invention or a pharmaceutically acceptable form thereof or a pharmaceutical composition of the present invention can achieve at least one of the following effects: (1) preventing the occurrence of a disease in an animal that may be predisposed to the disease but has not yet experienced or shown disease pathology or symptomatology; (2) inhibiting the disease (i.e., preventing further development of the pathology and / or symptomatology) in an animal that is experiencing or showing disease pathology or symptomatology; (3) improving the disease (i.e., reversing the pathology and / or symptomatology) in an animal that is experiencing or showing disease pathology or symptomatology.

[0248] The term "administer (administrate / administrating / administration)" (or "administering a drug") refers to the process of applying an active pharmaceutical ingredient (such as a compound of the present invention) or a pharmaceutical composition containing an active pharmaceutical ingredient (such as the pharmaceutical composition of the present invention) to an individual or to a site of its cells, tissues, organs, biological fluids, etc., so that the active pharmaceutical ingredient or the pharmaceutical composition comes into contact with the individual or the site of its cells, tissues, organs, biological fluids, etc. Common administration methods include (but are not limited to) oral administration, subcutaneous administration, intramuscular administration, subperitoneal administration, ocular administration, nasal administration, sublingual administration, rectal administration, vaginal administration, etc.

[0249] The term "in need thereof" refers to a judgment by a doctor or other caregiver that an individual needs or will benefit from a preventive and / or therapeutic process, which judgment is based on various factors within the doctor's or other caregiver's area of expertise.

[0250] The term "individual" (or "subject") refers to a human or non-human animal. The individuals of the present invention include individuals (patients) suffering from diseases and / or disorders and normal individuals. The non-human animals of the present invention include all vertebrates, such as non-mammals, such as birds, amphibians, reptiles, etc., and mammals, such as non-human primates, domestic animals and / or domesticated animals (such as sheep, dogs, cats, cows, pigs, etc.).

[0251] In order to make the objectives and technical solutions of the present invention clearer, the following describes the implementation schemes of the present invention in detail with reference to embodiments. However, those skilled in the art will understand that the following embodiments are only used to illustrate the present invention and should not be construed as limiting the scope of the present invention.

[0252] For the reagents or instruments used in the examples, if the manufacturer is not indicated, they are all conventional products that can be obtained through commercial purchase. For those without specific conditions indicated, all are carried out under conventional conditions or conditions recommended by the manufacturer. The term "room temperature" used in the present invention refers to 20°C ± 5°C. When used to modify a certain numerical value or numerical range, the term "about" used in the present invention means including the numerical value or numerical range and the error range acceptable to those skilled in the art for the numerical value or numerical range, for example, the error range is ±10%, ±5%, ±4%, ±3%, ±2%, ±1%, ±0.5%, etc.

[0253] In the conventional synthesis methods, as well as in the examples and intermediate synthesis examples, the meanings of the respective abbreviations are shown in the following table.

[0254] Abbreviation Meaning Abbreviation Meaning DMF N,N-Dimethylformamide Boc tert-Butyloxycarbonyl Tf Trifluoromethanesulfonyl THF Tetrahydrofuran DIAD Diisopropyl Azodicarboxylate DEAD Diethyl Azodicarboxylate DCM Dichloromethane DMSO Dimethyl Sulfoxide DIPEA N,N-Diisopropylethylamine <![CDATA[Pd2(dba)3]]> Tris(dibenzylideneacetone)dipalladium(0) <![CDATA[Pd(PPh3)2Cl2]]> Bis(triphenylphosphine)palladium(II) Dichloride <![CDATA[Pd(dppf)Cl2]]> [1,1'-Bis(diphenylphosphino)ferrocene]palladium(II) Dichloride h hour <![CDATA[PPh3]]> Triphenylphosphine ADDP Dipiperidyl Azodicarboxylate Tol Toluene <![CDATA[Et2O]]> Diethyl Ether n-BuLi n-Butyllithium t-BuLi tert-Butyllithium LiHMDS Lithium Bis(trimethylsilyl)amide NaHMDS Sodium Bis(trimethylsilyl)amide EA Ethyl Acetate ACN Acetonitrile <![CDATA[Et3N]]> Triethylamine t-BuOK Potassium tert-Butoxide MeONa Sodium Methoxide Diox 1,4-Dioxane <![CDATA[Pd(PPh3)4]]> Tetrakis(triphenylphosphine)palladium(0) <![CDATA[Pd(OAc)2]]> Palladium(II) Acetate DIBAL Diisobutylaluminum Hydride DCSO Thionyl Chloride

[0255] The structures of the compounds described in the following examples were determined by nuclear magnetic resonance ( 1Determined by 1H-NMR) and / or mass spectrometry (MS).

[0256] Nuclear magnetic resonance ( 1 For the determination instrument of 1H-NMR, a Bruker 400 MHz nuclear magnetic resonance instrument is used. The solvents for determination are deuterated methanol (CD3OD), deuterated chloroform (CDCl3), hexadeuterated dimethyl sulfoxide (DMSO-d6), or deuterated water (D2O), and the internal standard substance is tetramethylsilane (TMS).

[0257] The meanings represented by the abbreviations in the nuclear magnetic resonance (NMR) data in the following examples are as follows:

[0258] s: singlet, d: doublet, t: triplet, q: quartet, dd: double doublet, qd: quartet doublet, ddd: double double doublet, ddt: double double triplet, dddd: double double double doublet, m: multiplet, br: broad peak, J: coupling constant, Hz: Hertz, δ: chemical shift.

[0259] All chemical shift (δ) values are given in units of parts per million (ppm).

[0260] For the determination instrument of mass spectrometry (MS), an Agilent 6120B mass spectrometer is used, and the ion source is an electrospray ionization source (ESI).

[0261] The examples of the present invention are purified by preparative thin layer chromatography (Prep-TLC) using the following method:

[0262] Use a thin layer chromatography silica gel plate produced by Yantai Xin Nuo Chemical Co., Ltd. (thickness is 1 mm, specification is 200×200 mm, and the binder is sodium carboxymethyl cellulose). The detection instrument is a ZF-20D dark box type ultraviolet analyzer, and the detection wavelengths are 254 nm and / or 365 nm.

[0263] The examples of the present invention are purified by preparative high performance liquid chromatography (Prep-HPLC) using the following method:

[0264] Method A:

[0265] Chromatographic column: Waters SunFire Prep C18 OBD 5μm 19×150 mm;

[0266] Mobile phase A: acetonitrile; Mobile phase B: water (containing 0.05 wt% formic acid);

[0267] Time (min) Mobile Phase A (v%) Mobile Phase B (v%) Flow Rate (mL / min) 0.00 10.0 90.0 28 18.00 90.0 10.0 28

[0268] Method B:

[0269] Chromatographic column: Waters SunFire Prep C18 OBD 5μm 19×150mm;

[0270] Mobile phase A: acetonitrile; Mobile phase B: water (containing 0.05 wt% ammonium bicarbonate);

[0271] Time (min) Mobile Phase A (v%) Mobile Phase B (v%) Flow Rate (mL / min) 0.00 10.0 90.0 28 18.00 90.0 10.0 28

[0272] [Compound Preparation and Identification]

[0273] Example 1: Synthesis of 3-((3-(5-chloro-3-methyl-2-(piperidin-4-yloxy)phenyl)-2-methyl-2H-thieno[3,2-c]pyrazol-5-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 1).

[0274]

[0275] Step 1: Synthesis of tert-butyl 4-(2-bromo-4-chloro-6-methylphenoxy)piperidine-1-carboxylate (Compound 1-2):

[0276] Compound 1-1 (8.80 g, 39.75 mmol), tert-butyl 4-hydroxypiperidine-1-carboxylate (4.00 g, 19.87 mmol), PPh3 (10.43 g, 39.75 mmol), and dry THF (240 mL) were successively added to the reaction flask. At 0 °C, DIAD (7.8 mL, 39.75 mmol) was added dropwise. After the addition was complete, the reaction mixture was slowly warmed to room temperature and reacted for 16 h. The reaction solution was concentrated under reduced pressure and purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 23 / 2, v / v) to obtain 5.45 g of the title compound.

[0277] ESI-MS: m / z 347.8, 349.8 [M-56+H] + 。

[0278] Step 2: Synthesis of tert-butyl 4-(4-chloro-2-methyl-6-(tributylstannyl)phenoxy)piperidine-1-carboxylate (Compound 1-3):

[0279] At -40 °C, a solution of n-BuLi in n-hexane (2.5 mol / L, 7.41 mL) was added dropwise to a solution of compound 1-2 (5.00 g, 12.35 mmol) in Tol (50 mL). After reacting for 0.5 h, tributyltin chloride (4.02 mL, 14.82 mmol) was added dropwise. After the addition was complete, the reaction mixture was slowly warmed to 0 °C and reacted for 3 h. The reaction was quenched by adding water (20 mL), and the mixture was extracted with EA (3 × 25 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. After purification by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 20 / 1, v / v), 6.31 g of the title compound was obtained.

[0280] 1 1H-NMR (400 MHz, CDCl3): δ 7.16 - 7.04 (m, 2H), 4.11 (brs, 2H), 3.99 - 3.90 (m, 1H), 2.69 (t, J = 11.9 Hz, 2H), 2.23 (s, 3H), 1.89 (d, J = 11.2 Hz, 2H), 1.59 - 1.48 (m, 7H), 1.46 (s, 9H), 1.45 - 1.39 (m, 1H), 1.32 (h, J = 7.3 Hz, 6H), 1.17 - 0.98 (m, 6H), 0.89 (t, J = 7.3 Hz, 9H).

[0281] Step 3: Synthesis of methyl 3-iodo-2-methyl-2H-thieno[3,2-c]pyrazole-5-carboxylate (Compound 1-5):

[0282] Compound 1-4 (1.00 g, 3.25 mmol) was added to methanol (30 mL). At room temperature, a solution of MeONa in methanol (1 mL, 33 wt%) was added dropwise, and the mixture was reacted for 0.5 h. Then, methyl iodide (0.61 mL, 9.74 mmol) was added, and after reacting for 2 h, an additional portion of methyl iodide (0.61 mL, 9.74 mmol) was added, and the reaction was continued for 2 h. The reaction solution was concentrated under reduced pressure and purified by silica gel column chromatography (eluent: dichloromethane = 100%, v), and 484 mg of the title compound was obtained.

[0283] 1 1H-NMR (400 MHz, CDCl3): δ 7.89 (s, 1H), 4.13 (s, 3H), 3.92 (s, 3H).

[0284] ESI-MS: m / z 322.9 [M + H] + 。

[0285] Step 4: Synthesis of methyl 3-(2-(1-(tert-butoxycarbonyl)piperidin-4-yloxy)-5-chloro-3-methylphenyl)-2-methyl-2H-thieno[3,2-c]pyrazole-5-carboxylate (Compound 1-6):

[0286] To a reaction flask were successively added Compound 1-5 (350 mg, 1.09 mmol), Compound 1-3 (802 mg, 1.30 mmol), CuI (414 mg, 2.17 mmol), and Diox (17.5 mL). Under nitrogen protection, Pd(PPh3)4 (251 mg, 0.22 mmol) was added, and the temperature was raised to 110 °C for reaction for 8 h. Heating was stopped and the mixture was cooled to room temperature. After filtration, the filtrate was concentrated under reduced pressure and then purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 3 / 1, v / v) to obtain 466 mg of the title compound.

[0287] ESI-MS: m / z 519.8, 521.8 [M+H] + 。

[0288] Step 5: Synthesis of tert-butyl 4-(4-chloro-2-(5-(hydroxymethyl)-2-methyl-2H-thieno[3,2-c]pyrazol-3-yl)-6-methylphenoxy)piperidine-1-carboxylate (Compound 1-7):

[0289] Compound 1-6 (400 mg, 0.77 mmol) was dissolved in THF (10 mL), and a THF solution of LiAlH4 (0.6 mL, 1.54 mmol) was added dropwise at 0 °C. After the addition was complete, the reaction was continued for 1.5 h. The reaction was quenched by adding water (1 mL), and the reaction solution was concentrated under reduced pressure and then purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 2 / 1, v / v) to obtain 314 mg of the title compound.

[0290] ESI-MS: m / z 491.8, 493.8 [M+H] + 。

[0291] Step 6: Synthesis of tert-butyl 4-(4-chloro-2-(5-(chloromethyl)-2-methyl-2H-thieno[3,2-c]pyrazol-3-yl)-6-methylphenoxy)piperidine-1-carboxylate (Compound 1-8):

[0292] Compound 1-7 (150 mg, 0.30 mmol) was dissolved in DCM (3 mL), cooled to 0 °C, and then DCSO (0.04 mL, 0.61 mmol) was added. The mixture was stirred for an additional 0.5 h. The reaction solution was concentrated under reduced pressure to obtain 150 mg of the title compound.

[0293] ESI-MS: m / z 509.6, 511.7 [M+H]+ 。

[0294] Step 7: Synthesis of tert-butyl 4-(4-chloro-2-(5-((6,6-dimethyl-2,4-dioxo-3-azabicyclo[3.1.0]hexan-3-yl)methyl)-2-methyl-2H-thieno[3,2-c]pyrazol-3-yl)-6-methylphenoxy)piperidine-1-carboxylate (Compound 1-9):

[0295] Dissolve 6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (82 mg, 0.59 mmol) in DMF (1.5 mL). After cooling to 0 °C, add NaH (23 mg, 0.59 mmol, 60%), and react for 0.5 h. Then dropwise add a solution of Compound 1-8 (150 mg, 0.29 mmol) in DMF (1.5 mL). After the addition is complete, raise the temperature to room temperature and react for 4.5 h. Quench the reaction with water, dilute with EA (2 mL), wash with water (3 × 5 mL), dry the organic phase over anhydrous sodium sulfate, concentrate under reduced pressure, purify by preparative high performance liquid chromatography (Method A), concentrate the preparation under reduced pressure, and freeze-dry to obtain 129 mg of the title compound.

[0296] ESI-MS: m / z 612.8, 614.8 [M+H] + 。

[0297] Step 8: Synthesis of hydrochloride of 3-((3-(5-chloro-3-methyl-2-(piperidin-4-yloxy)phenyl)-2-methyl-2H-thieno[3,2-c]pyrazol-5-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 1):

[0298] Add Compound 1-9 (100 mg, 0.13 mmol) to a reaction flask, then add hydrochloric acid / ethyl acetate solution (2 mL, 4 mol / L), and react at room temperature for 1 h. Concentrate the reaction solution under reduced pressure, separate and purify by preparative high performance liquid chromatography (Method A), concentrate the preparation under reduced pressure, add hydrochloric acid (0.5 mL, 1 mol / L) to the preparation, and freeze-dry to obtain 68 mg of the hydrochloride of the title compound.

[0299] 11H-NMR (400 MHz, DMSO-d6): δ 8.68 (brs, 1H), 8.33 (brs, 1H), 7.52 (d, J = 2.5 Hz, 1H), 7.41 (d, J = 2.6 Hz, 1H), 7.10 (s, 1H), 4.65 (s, 2H), 3.90 (s, 3H), 3.79 (dt, J = 7.7, 4.1 Hz, 1H), 2.87 - 2.60 (m, 4H), 2.57 (s, 2H), 2.34 (s, 3H), 1.73 - 1.59 (m, 2H), 1.44 - 1.28 (m, 2H), 1.16 (s, 3H), 0.99 (s, 3H).

[0300] ESI-MS: m / z 513.0, 515.0 [M+H] + 。

[0301] Example 2: Synthesis of 3-((3-(5-chloro-3-methyl-2-(1-methylpiperidin-4-yloxy)phenyl)-2-methyl-2H-thieno[3,2-c]pyrazol-5-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 8).

[0302]

[0303] Add the hydrochloride salt of Compound 1 (26 mg, 0.05 mmol) to a reaction flask and dissolve it in acetonitrile (1 mL). Dropwise add aqueous formaldehyde solution (0.05 mL, 37 - 40% purity). After reacting at room temperature for 20 minutes, add sodium cyanoborohydride (9 mg, 0.14 mmol) and react at room temperature for 3 h. Quench the reaction with saturated aqueous sodium bicarbonate solution, extract with EA (3 × 2 mL), combine the organic phases, dry over anhydrous sodium sulfate, filter, concentrate the filtrate, and then separate and purify by preparative high performance liquid chromatography (Method B). Freeze-dry the preparation solution to obtain 14 mg of the title compound.

[0304] 1 1H-NMR (400 MHz, CD3OD): δ 7.38 (d, J = 2.4 Hz, 1H), 7.22 (d, J = 2.6 Hz, 1H), 7.03 (s, 1H), 4.69 (s, 2H), 3.88 (s, 3H), 3.54 (brs, 1H), 2.44 (s, 2H), 2.34 (s, 3H), 2.33 - 2.11 (m, 2H), 2.06 (s, 3H), 1.89 (brs, 2H), 1.51 (brs, 2H), 1.43 (brs, 2H), 1.20 (s, 3H), 1.04 (s, 3H).

[0305] ESI-MS: m / z 527.2, 529.1 [M+H] + 。

[0306] [Pharmacological Activity Test]

[0307] Test Example 1: In vitro enzymatic activity inhibition test of USP7 (protease).

[0308] 1. Test system:

[0309] Protease: His6-USP7 (Boston Biochem);

[0310] Substrate: Ubiquitin-Rhodamine 110 (Boston Biochem);

[0311] Buffer: 5×USP7 assay buffer (BPS).

[0312] 2. Test parameters:

[0313] USP7 concentration: 3 nM;

[0314] Ubiquitin-Rhodamine 110 concentration: 100 nM;

[0315] Buffer system: 1.25×USP7 assay buffer; 0.06% BSA; 1 mM DTT; ddH2O;

[0316] Reaction temperature: 25°C;

[0317] Incubation time of compound and enzyme: 20 min;

[0318] Enzyme kinetic reaction time: 20 min;

[0319] Microplate reader parameters: BMG PHERAstar FS fluorescence microplate reader, excitation wavelength 485 nm, emission wavelength 520 nm.

[0320] 3. Test method:

[0321] Conduct the test according to the kit instructions. The steps are as follows:

[0322] Test group: Incubate the mixture of the test compound and protease USP7 in the buffer system at 25°C for 20 min, add the substrate Ubiquitin-Rhodamine 110 to initiate the reaction, and use the enzyme kinetic method to read the fluorescence values of each well.

[0323] Negative group: Replace the test compound with 0.2% DMSO aqueous solution, and the experimental method is the same as that of the test group.

[0324] Blank group: Replace the test compound with 0.2% DMSO aqueous solution and do not add protease USP7. The experimental method is the same as that of the test group.

[0325] 4. Data processing:

[0326] Calculate the relative inhibitory activity of each concentration group. Inhibition rate (%) = 100% - (fluorescence value of the test group - fluorescence value of the blank group) / (fluorescence value of the negative group - fluorescence value of the blank group) × 100%. Fit the curve according to the four-parameter model and calculate the half-maximal inhibitory concentration (IC 50 ).

[0327] 5. Test results:

[0328] Determine the inhibition of the compound of the present invention on USP7 activity according to the above method. The results are shown in Table 1.

[0329] Table 1. Results of USP7 enzyme activity inhibition test

[0330] Example Number <![CDATA[IC 50 (nM)]]> 1 13.39±0.04 2 36.26±2.52

[0331] In the USP7 enzyme activity inhibition test, the compound of the present invention exhibits extremely strong inhibitory activity.

[0332] Except for those embodiments described herein, various modifications of the present invention will be apparent to those skilled in the art in light of the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. All references cited in the present invention (including all patents, patent applications, journal articles, books, and any other publications) are incorporated herein by reference in their entirety.

Claims

1. A compound having the structure of formula I or a pharmaceutically acceptable salt thereof, wherein, R 0 is hydrogen; L 1 is methylene; R 1 For R 2 and R 4 is hydrogen; R 3 is chlorine; R 5 and R 6 are methyl; L 2 Selected from -O-, -NH-, -C(O)-, -O-CH2- and -CH2-; A is C 4-5 a cycloalkyl group or a 4- to 8-membered heterocycloalkane ring; Each R 7 and R 8 are each independently selected from hydrogen, amino, C 1-6 alkyl, and C 1-6 alkoxy; z 1 and z 2 are each independently selected from 0, 1, and 2.

2. The following compounds or pharmaceutically acceptable salts thereof:

3. A pharmaceutical composition comprising the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 or 2, and one or more pharmaceutically acceptable carriers.

4. A medicament comprising: a) The compound or a pharmaceutically acceptable salt thereof according to claim 1 or 2 or the pharmaceutical composition according to claim 3; b) Optionally present packaging and / or instructions for use.

5. Use of the compound or a pharmaceutically acceptable salt thereof according to claim 1 or 2 or the pharmaceutical composition according to claim 3 or the medicament according to claim 4 in the manufacture of a medicament for the prevention and / or treatment of a disease or disorder mediated at least in part by UPS7.

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