Five-membered and six-membered nitrogen-containing compound as well as preparation method and application thereof

Abstract

The invention discloses a five-membered and six-membered nitrogen-containing compound as well as a preparation method and application thereof. The compound provided by the invention has relatively good PARG inhibitory activity.

Description

Technical Field

[0001] The present invention relates to a five - membered and six - membered nitrogen - containing compound, a preparation method thereof, and an application thereof. Background Art

[0002] PARG is a poly(ADP - ribose) hydrolase encoded by a single gene and the first to be identified (Mirella et al. Human poly(ADP - ribose) glycohydrolase is expressed in alternative splice variants yielding isoforms that localize to different cell compartments. Experimental Cell Research, 2004, 297(2): 521 - 532.), which can hydrolyze the O - glycosidic bond between ADP - ribose subunits. PARG consists of four domains (Meyer B et al. Clustered DNA damage induces pan - nuclear H2AX phosphorylation mediated by ATM and DNA - PK. Nucleic Acids Res, 2003, 41: 6109 - 6118.; O’Sullivan J. et al. Emerging roles of erase enzymes in the dynamic control of protein ADP - ribosylation. Nature Communication, 2019, 10, 1182.), including an N - terminal disordered domain, a linker domain, a C - terminal catalytic domain, and an ADP - ribose binding domain. Among them, the N - terminal non - conserved disordered domain is not necessary for in vitro activity, and the C - terminal ADP - ribose binding domain is highly conserved with other Macro - domains and together with the catalytic domain forms the smallest structure with complete enzyme activity in vitro (Slade D et al. The structure and catalytic mechanism of a poly(ADP - ribose) glycohydrolase. Nature, 2011, 477: 616 - 620.).

[0003] Poly(ADP-ribosyl)ation (PARylation) is a unique post-translational modification that plays an important role in maintaining genomic stability in different signaling pathways, especially in DNA damage repair. Poly(ADP-ribose), abbreviated as PAR, consists of adenosine diphosphate-ribose units, is formed by poly(ADP-ribose) polymerases (PARPs) using NAD+ as a donor, and can be rapidly degraded by polyribosylhydrolases. After DNA damage, many DNA damage response factors recognize PARylation and are recruited to the vicinity of DNA damage sites through PARylation. However, PARylation needs to be digested in a timely manner so that DNA damage response factors can directly recognize DNA damage and play a repair function. Otherwise, DNA repair factors will be trapped near DNA damage by PARylation and cannot function properly in repair. Therefore, dePARylation, as the direct downstream step of PARylation in DNA repair, inhibiting this process will affect PARylation-dependent DNA damage repair and selectively kill tumor cells with DNA repair defects.

[0004] As the most important poly(ADP-ribose) hydrolase, PARG acts on approximately 90% of the PAR in cells (Laetitia D et al. Poly(ADP-ribose) glycohydrolase (PARG) and its therapeutic potential. Front Biosci, 2009, 14(5): 1619 - 1626.), and plays a role in multiple cellular processes such as DNA damage repair, DNA replication, chromatin regulation, transcription, and apoptosis. Imbalance of intracellular PAR levels caused by abnormal PARG function will affect the transformation and invasion of cancer cells (Rack J D et al. Macrodomain: structure, function, evolution, and catalytic activities. Annu Rev Biochem. 2016, 85: 431 - 454; Marques M et al. Oncogenic activity of poly(ADP-ribose) glycohydrolase. Oncogene, 2019, 38: 2177 - 2191).

[0005] PARG has been proven to be associated with various diseases. Data indicate (Maud M et al, Oncogenic activity of poly(ADP-ribose) glycohydrolase. Oncogene, 2019, 38: 2177-2191.) that elevated PARG is associated with poor prognosis in HER2+ patients. PARG and HER2 synergistically promote the growth of breast cancer cells, and inhibiting PARG significantly affects the growth and migration of breast cancer. Additionally, research shows (Mincheng Y et al. PARG inhibition limits HCC progression and potentiates the efficacy of immune checkpoint therapy. Hepatic and Biliary Cancer, 2022, S0168-8278(22)00072-1.) that high expression of PARG is closely associated with poor prognosis in liver cancer, and hepatocyte-specific PARG loss-of-function affects tumorigenesis. Therefore, PARG inhibitors have received increasing attention as potential therapeutic means. There are currently multiple PARG inhibitors under development, and all are in the preclinical research stage. Therefore, developing small molecule drugs that can target and inhibit PARG activity and providing safer and more effective PARG inhibitors for patients is of great research significance. Summary of the Invention

[0006] The technical problem to be solved by the present invention is to overcome the relatively single structure of existing PARG inhibitor compounds. To this end, the present invention provides a five-membered fused six-membered nitrogen-containing compound, its preparation method and application. The compounds of the present invention have good PARG inhibitory activity.

[0007] The present invention provides any one of the following compounds, its pharmaceutically acceptable salt or isotopic compound:

[0008]

[0009] The present invention also provides a pharmaceutical composition, which comprises substance Z and pharmaceutical excipients, and the substance Z is any one of the above-mentioned compounds, its pharmaceutically acceptable salt or isotopic compound.

[0010] The present invention also provides the application of substance Z in the preparation of PARG inhibitors and drugs for treating and / or preventing PARG-related diseases, and the substance Z is any one of the above-mentioned compounds, its pharmaceutically acceptable salt or isotopic compound.

[0011] In a preferred embodiment, the PARG-related disease is breast cancer, such as triple-negative breast cancer, and other PARG-related diseases.

[0012] The present invention also provides the use of substance Z in the preparation of a medicament for treating and / or preventing breast cancer, wherein substance Z is any one of the above-mentioned compounds, its pharmaceutically acceptable salts or isotopic compounds.

[0013] In a preferred embodiment, the breast cancer is triple-negative breast cancer.

[0014] The present invention also provides a method for treating and / or preventing PARG-related diseases, which comprises administering an effective amount of substance Z to a patient, wherein substance Z is any one of the above-mentioned compounds, its pharmaceutically acceptable salts or isotopic compounds.

[0015] In a preferred embodiment, the PARG-related disease is breast cancer, such as triple-negative breast cancer, and other PARG-related diseases.

[0016] The present invention also provides a substance Z for treating and / or preventing PARG-related diseases, wherein substance Z is any one of the above-mentioned compounds, its pharmaceutically acceptable salts or isotopic compounds.

[0017] In a preferred embodiment, the PARG-related disease is breast cancer, such as triple-negative breast cancer, and other PARG-related diseases.

[0018] Glossary of Terms

[0019] The term "pharmaceutically acceptable" means relatively non-toxic, safe and suitable for use by patients.

[0020] The term "pharmaceutically acceptable salt" refers to a salt obtained by reacting a compound with a pharmaceutically acceptable acid or base. When a compound contains a relatively acidic functional group, a base addition salt can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable base in a suitable inert solvent. Pharmaceutically acceptable base addition salts include, but are not limited to: sodium salts, potassium salts, calcium salts, aluminum salts, magnesium salts, bismuth salts, ammonium salts, etc. When a compound contains a relatively basic functional group, an acid addition salt can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable acid in a suitable inert solvent. Pharmaceutically acceptable acid addition salts include, but are not limited to: hydrochloride salts, sulfate salts, formate salts, methanesulfonate salts, etc. For details, see Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl, Camille G. Wermuth, 2011, 2nd Revised Edition).

[0021] The term "isotope compound" refers to a compound in which the isotope abundances of one or more atoms are different from their natural abundances. For example, one or more atoms in the compound are replaced by atoms with a lower mass number in nature - one hydrogen atom in the compound is replaced by deuterium, or C is 13 replaced by C.

[0022] Based on the common knowledge in this field, the above preferred conditions can be combined arbitrarily to obtain the preferred examples of the present invention.

[0023] The reagents and raw materials used in the present invention are all commercially available.

[0024] The positive and progressive effects of the present invention are as follows: the compounds of the present invention have good PARG inhibitory activity. Detailed implementation manners

[0025] All compounds of the present invention can be synthesized by those skilled in the art of organic chemistry through different methods. The general synthetic schemes for preparing the compounds of the present invention are described below. These schemes are general and do not mean to limit the possible techniques that those skilled in the art can use to prepare the compounds disclosed herein. Different methods for preparing the compounds of the present invention are obvious to those skilled in the art. In addition, the various steps in the synthesis can be carried out in an alternating order to obtain the desired one or more compounds. The preparation and examples described below give examples of preparing the compounds of the present invention by the methods described in the general schemes. The preparation of compounds in the examples containing chiral centers can be carried out by techniques mastered by those skilled in the art. For example, chiral compounds can be prepared by chiral resolution of racemic products through HPLC; or, the example compounds can be prepared by known methods to obtain chiral compounds.

[0026] The chemical reactions and synthetic techniques described herein were carried out in the reagents and corresponding solvents described in the text, and the corresponding reaction yields were also affected by the reagents and solvents used. In addition, it should be understood that in the synthetic methods described below, all mentioned reaction conditions, including the choice of solvent, reaction atmosphere, reaction temperature, experimental duration, and reaction feeding sequence, should be regarded as the standard operating conditions for the reaction, which should be easily recognizable by those skilled in the art. At the same time, it is also understandable to those skilled in the field of organic synthesis. The functional groups present on each part of the molecule must be compatible with the reagents used and the reaction itself. For the limitation that some of the functional groups present on each part of the molecule are incompatible with the reaction conditions, alternative methods must be used, which will be obvious to those skilled in the art. It is obvious for those skilled in the art to judge and adjust the order of the synthesis steps, or select a specific synthetic process scheme to obtain the compounds required by the present invention. This is understandable and easily recognizable by those skilled in the field of organic synthesis. It should also be recognized that another major consideration in designing any synthetic route in this field is the reasonable selection of protecting groups to tolerate the reactive functional groups present in the compounds described in the present invention. Specifically, reference can be made to (Protective Groups in Organic Synthesis, Third Edition, Wiley and Sons (1999)) written by authorities in the field of chemistry such as Greene et al.

[0027] Examples

[0028] The preparation of compounds and the intermediates used in the preparation of compounds can be prepared using the procedures shown in the following examples and related procedures. The methods and conditions used in these examples and the actual compounds prepared in these examples are not meant to be limiting, but rather to illustrate how to prepare the relevant compounds. The starting materials and reagents used in these examples, when not prepared by the procedures described herein, are generally commercially available, or reported in relevant chemical literature, or can be prepared by using the procedures described in the chemical literature.

[0029] In the examples given in this article, the term "drying and concentration" generally refers to adding anhydrous sodium sulfate or magnesium sulfate to dry the solution in an organic solvent, followed by filtration and removal of the solvent from the filtrate (usually carried out under reduced pressure and at a temperature suitable for the stability of the prepared compound). Column chromatography is usually carried out using conventional column chromatography or flash column chromatography for column separation and purification, or using a pre-packed silica gel column on a medium-pressure chromatograph (Biotage Isola One) and eluting with a specified solvent or solvent mixture. In some cases, a 20 cm x 20 cm x 0.5 mm or 20 cm x 20 cm x 1 mm silica gel plate is used in an appropriate solvent system to rapidly purify the final product by preparative thin-layer chromatography. Preparative high-performance liquid chromatography (HPLC) is carried out using a reversed-phase column (Waters Sunfire C18, Waters Xbridge C18 or a similar reversed-phase column) suitable for the amount of the compound to be separated, usually eluting with a gradient of the concentration of methanol or acetonitrile added to the aqueous phase, and the eluent contains 0.05% or 0.1% formic acid, trifluoroacetic acid or 10 mM ammonium acetate, and the elution rate matches the size of the reversed-phase column used and the resolution of the prepared product.

[0030] List of Abbreviations

[0031]

[0032]

[0033] Intermediate 1: 1,8-Dichloro-N-(3-methyloxetan-3-yl)-3-(5-(trifluoromethyl)-1,3,4-thiadiazolo -2-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)imidazo[1,5-a]pyridine-6-sulfonamide

[0034]

[0035] Step 1: 6-Bromo-8-chloro-N'-(2,2,2-trifluoroacetyl)imidazo[1,5-a]pyridine-3-carbohydrazide

[0036] Under nitrogen protection, trifluoroacetic anhydride (435 mg, 2.07 mmol) was added to a solution of 6-bromo-8-chloroimidazo[1,5-a]pyridine-3-carbohydrazide (600 mg, 2.07 mmol) in dichloromethane (6 mL). The reaction mixture was stirred at room temperature for 30 minutes. After the reaction was complete, water was added and the mixture was extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0 - 15% ethyl acetate) to obtain the title compound (yellow solid, 600 mg, yield 75.1%). LC / MS (ESI) (m / z): 385 [M+H] + .

[0037] Step 2: 2-(6-Bromo-8-chloroimidazo[1,5-a]pyridin-3-yl)-5-(trifluoromethyl)-1,3,4-thiadiazole

[0038] Under nitrogen protection, Lawesson's reagent (692 mg, 1.71 mmol) was added to a solution of 6-bromo-8-chloro-N'-(2,2,2-trifluoroacetyl)imidazo[1,5-a]pyridine-3-carbohydrazide (600 mg, 1.56 mmol) in anhydrous toluene (6 mL). The reaction mixture was stirred at 110 °C for 4 h. After completion of the reaction, sodium bicarbonate was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0 - 20% ethyl acetate) to obtain the title compound (yellow solid, 500 mg, yield 83.7%). LC / MS (ESI) (m / z): 383 [M+H] + .

[0039] Step 3: 2-(6-(Benzylthio)-8-chloroimidazo[1,5-a]pyridin-3-yl)-5-(trifluoromethyl)-1,3,4-thiadiazole

[0040] Under nitrogen protection, N,N-diisopropylethylamine (505 mg, 3.91 mmol), Xantphos (151 mg, 0.26 mmol), and Pd2(dba)3 (119 mg, 0.13 mmol) were successively added to a solution of 2-(6-bromo-8-chloroimidazo[1,5-a]pyridin-3-yl)-5-(trifluoromethyl)-1,3,4-thiadiazole (500 mg, 1.30 mmol) and benzyl mercaptan (0.15 mL, 1.30 mmol) in 1,4-dioxane (5 mL). The reaction mixture was purged with nitrogen three times and stirred at 80 °C for 3 h under nitrogen protection. After completion of the reaction, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0 - 25% ethyl acetate) to obtain the title compound (yellow solid, 510 mg, yield 91.6%). LC / MS (ESI) (m / z): 427 [M+H] + .

[0041] Step 4: 1,8-Dichloro-3-(5-(trifluoromethyl)-1,3,4-thiadiazol-2-yl)imidazo[1,5-a]pyridine-6- sulfonyl chloride

[0042] Under nitrogen protection, sulfuryl chloride (13.5 g, 100 mmol) was slowly added dropwise to a solution of 2-(6-(benzylthio)-8-chloroimidazo[1,5-a]pyridin-3-yl)-5-(trifluoromethyl)-1,3,4-thiadiazole (7.0 g, 16.4 mmol) in dichloromethane (150 mL). The reaction mixture was stirred at 0 °C for 60 minutes under nitrogen. After the reaction was complete, it was slowly added to a saturated sodium bicarbonate solution on ice, and the layers were separated. The aqueous phase was extracted with dichloromethane. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the title compound (yellow solid, 7.0 g, yield 97.5%). LC / MS (ESI) (m / z): 437 [M+H] + .

[0043] Step 5: 1,8-Dichloro-N-(3-methyloxetan-3-yl)-3-(5-(trifluoromethyl)-1,3,4-thiadiazolo -2-yl)imidazo[1,5-a]pyridine-6-sulfonamide

[0044] Under nitrogen protection, a solution of 1,8-dichloro-3-(5-(trifluoromethyl)-1,3,4-thiadiazol-2-yl)imidazo[1,5-a]pyridine-6-sulfonyl chloride (7.0 g, 16.0 mmol) in dichloromethane (100 mL) was slowly added dropwise to a mixture of 3-methyloxetane-3-amine (2.8 g, 32.0 mmol) and pyridine (3 mL) in dichloromethane (100 mL). The reaction mixture was stirred at 0 °C for 60 minutes under nitrogen. After the reaction was complete, it was slowly added to ice water, and the layers were separated. The aqueous phase was extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0 - 45% ethyl acetate) to obtain the title compound (yellow solid, 4.9 g, yield 62.7%). LC / MS (ESI) (m / z): 488 [M+H] + .

[0045] Step 6: 1,8-Dichloro-N-(3-methyloxetan-3-yl)-3-(5-(trifluoromethyl)-1,3,4-thiadiazolo -2-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)imidazo[1,5-a]pyridine-6-sulfonamide

[0046] Under an ice bath, 2-(trimethylsilyl)ethoxymethyl chloride (3.4 g, 20 mmol) was slowly added dropwise to a solution of 1,8-dichloro-N-(3-methyloxetan-3-yl)-3-(5-(trifluoromethyl)-1,3,4-thiadiazol-2-yl)imidazo[1,5-a]pyridine-6-sulfonamide (4.9 g, 10 mmol) and N,N-diisopropylethylamine (3.9 g, 30 mmol) in dichloromethane (50 mL). The reaction mixture was stirred at room temperature for 2 hours. After the reaction was complete, water was slowly added, and the layers were separated. The aqueous phase was extracted with dichloromethane. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0 - 25% ethyl acetate) to obtain the title compound (yellow solid, 5.5 g, yield 86.8%). LC / MS (ESI) (m / z): 618 [M+H] + .

[0047] Intermediate 2: 1,8-Dichloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-N-(3-methyloxetan -3-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)imidazo[1,5-a]pyridine-6-sulfonamide

[0048]

[0049] Referring to the synthesis method of Intermediate 1, the difference is that difluoroacetic anhydride is used instead of trifluoroacetic anhydride in Step 1 to obtain Intermediate 2. LC / MS (ESI) (m / z): 600 [M+H] + .

[0050] Intermediate 3: 1-Bromo-8-chloro-3-(5-(trifluoromethyl)-1,3,4-thiadiazol-2-yl)imidazo[1,5-a]pyridine-6-sulfonyl chloride

[0051]

[0052] Step 1: 6-Bromo-8-chloro-N'-(2,2,2-trifluoroacetyl)imidazo[1,5-a]pyridine-3-carbohydrazide

[0053] Under nitrogen protection, trifluoroacetic anhydride (435 mg, 2.07 mmol) was added to a solution of 6-bromo-8-chloroimidazo[1,5-a]pyridine-3-carbohydrazide (600 mg, 2.07 mmol) in dichloromethane (6 mL). The reaction mixture was stirred at room temperature for 30 minutes. After the reaction was complete, water was added, and the mixture was extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0 - 15% ethyl acetate) to obtain the title compound (yellow solid, 600 mg, yield 75.1%). LC / MS (ESI) (m / z): 385 [M+H] + .

[0054] Step 2: 2-(6-Bromo-8-chloroimidazo[1,5-a]pyridin-3-yl)-5-(trifluoromethyl)-1,3,4-thiadiazole

[0055] Under nitrogen protection, Lawesson's reagent (692 mg, 1.71 mmol) was added to a solution of 6-bromo-8-chloro-N'-(2,2,2-trifluoroacetyl)imidazo[1,5-a]pyridine-3-carbohydrazide (600 mg, 1.56 mmol) in anhydrous toluene (6 mL). The reaction mixture was stirred at 110 °C for 4 hours. After the reaction was complete, sodium bicarbonate was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0 - 20% ethyl acetate) to obtain the title compound (yellow solid, 500 mg, yield 83.7%). LC / MS (ESI) (m / z): 383 [M+H] + .

[0056] Step 3: 2-(6-(Benzylthio)-8-chloroimidazo[1,5-a]pyridin-3-yl)-5-(trifluoromethyl)-1,3,4-thiadiazole

[0057] Under nitrogen protection, N,N-diisopropylethylamine (505 mg, 3.91 mmol), Xantphos (151 mg, 0.26 mmol), and Pd2(dba)3 (119 mg, 0.13 mmol) were successively added to a solution of 2-(6-bromo-8-chloroimidazo[1,5-a]pyridin-3-yl)-5-(trifluoromethyl)-1,3,4-thiadiazole (500 mg, 1.30 mmol) and benzyl mercaptan (0.15 mL, 1.30 mmol) in 1,4-dioxane (5 mL). The reaction mixture was purged with nitrogen three times and stirred at 80 °C for 3 hours under nitrogen protection. After the reaction was complete, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0 - 25% ethyl acetate) to obtain the title compound (yellow solid, 510 mg, yield 91.6%). LC / MS (ESI) (m / z): 427 [M+H] + .

[0058] Step 4: 2-(6-(Benzylthio)-1-bromo-8-chloroimidazo[1,5-a]pyridin-3-yl)-5-(trifluoromethyl)-1,3,4-thiadiazole

[0059] At 0 °C, NBS (213 mg, 1.20 mmol) was added to a solution of 2-(6-(benzylthio)-8-chloroimidazo[1,5-a]pyridin-3-yl)-5-(trifluoromethyl)-1,3,4-thiadiazole (510 mg, 1.20 mmol) in N,N-dimethylformamide (5 mL). The reaction mixture was stirred at room temperature for 30 minutes. After completion of the reaction, water was added and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0 - 25% ethyl acetate) to obtain the title compound (yellow solid, 550 mg, yield 91.2%). LC / MS (ESI) (m / z): 505 [M+H] + .

[0060] Step 5: 1-Bromo-8-chloro-3-(5-(trifluoromethyl)-1,3,4-thiadiazol-2-yl)imidazo[1,5-a]pyridine-6-sulfonyl chloride

[0061] At 0 °C, 1N hydrochloric acid (1.25 mL), water (1.25 mL) and dichlorohydantoin (643 mg, 3.27 mmol) were successively added to a solution of 2-(6-(benzylthio)-1-bromo-8-chloroimidazo[1,5-a]pyridin-3-yl)-5-(trifluoromethyl)-1,3,4-thiadiazole (550 mg, 1.10 mmol) in acetonitrile (5 mL). The reaction mixture was stirred at 0 °C for 10 minutes. After completion of the reaction, water was added and the mixture was extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was triturated with petroleum ether and dried in vacuo to obtain the title compound (yellow solid, 520 mg, yield 99.2%). LC / MS (ESI) (m / z): 481 [M+H] + .

[0062] Intermediate 4: 1-Bromo-8-chloro-3-[5-(difluoromethyl)-1,3,4-thiadiazol-2-yl]imidazo[1,5-a]pyridine-6-sulfonyl chloride

[0063]

[0064] With reference to the synthetic method of Intermediate 3, the difference is that difluoroacetic anhydride was used instead of trifluoroacetic anhydride in Step 1 to obtain Intermediate 4. LC / MS (ESI) (m / z): 462.7 [M+H] + .

[0065] Intermediate 5: 8-Chloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-N-(3-methyloxetan-3-yl)imidazo[1,5-a]pyridine-6-sulfonamide

[0066]

[0067] Step 1: 1-Bromo-8-chloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-N-(3-methyloxetan-3-yl)imidazo[1,5-a]pyridine-6-sulfonamide

[0068] At 0 °C, a solution of 1-bromo-8-chloro-3-[5-(difluoromethyl)-1,3,4-thiadiazol-2-yl]imidazo[1,5-a]pyridine-6-sulfonyl chloride (90 mg, 0.194 mmol) in DCM (2 mL) was added dropwise to a solution of 3-methyloxetan-3-amine (0.017 mL, 0.388 mmol) in pyridine (3 mL). The mixture was stirred at room temperature for 20 minutes. After the reaction was complete, water was added, and the mixture was extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0 - 50%) to obtain the title compound (yellow solid, 80 mg, yield 80.14%). LC / MS (ESI) (m / z): 514 / 516 [M+H] + .

[0069] Step 2: 8-Chloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-N-(3-methyloxetan-3-yl)imidazo[1,5-a]pyridine-6-sulfonamide

[0070] At room temperature, 10% Pd / C (20 mg, 0.188 mmol) was added to a solution of 1-bromo-8-chloro-3-[5-(difluoromethyl)-1,3,4-thiadiazol-2-yl]-N-(3-methyloxetan-3-yl)imidazo[1,5] solution-a]pyridine-6-sulfonamide (70 mg, 0.136 mmol) in EtOH (10 mL). The mixture was purged with hydrogen three times and stirred at room temperature under a hydrogen atmosphere for 6 hours. After the reaction was complete, it was filtered and concentrated to dryness. The residue was used directly in the next step without purification. LC / MS (ESI) (m / z): 436 [M+H] + 。

[0071] Intermediate 6: 8-Chloro-3-(5-(trifluoromethyl)-1,3,4-thiadiazol-2-yl)-N-(3-methyloxetan-3-yl)imidazo[1,5-a]pyridine-6-sulfonamide

[0072]

[0073] Referring to the synthesis method of Intermediate 5, except that the starting material is Intermediate 3, Intermediate 6 is obtained. LC / MS (ESI) (m / z): 453.9 [M+H] + .

[0074] Intermediate 7: 1-Bromo-8-chloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-N-(3-(fluoromethyl)oxetan-3-yl)imidazo[1,5-a]pyridine-6-sulfonamide

[0075]

[0076] Referring to the synthesis method of Step 1 of Intermediate 5, except that the starting material is 3-(fluoromethyl)oxetan-3-amine, Intermediate 7 is obtained. LC / MS (ESI) (m / z): 532 [M+H] + .

[0077] Example 1: (S)-1-Chloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-8-(3-(methoxymethyl) piperazin-1-yl)-N-(3-methyloxetan-3-yl)imidazo[1,5-a]pyridine-6-sulfonamide

[0078]

[0079] Step 1: (S)-4-(1-Chloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-6-(N-(3-methyloxetan -3-yl)-N-(2-(trimethylsilyl)ethoxy)methyl)sulfamoyl)imidazo[1,5-a]pyridin-8- yl)-2-(methoxymethyl)piperazine-1-carboxylic acid tert-butyl ester

[0080] Under nitrogen protection, cesium carbonate (1627 mg, 5.01 mmol), Ruphos (156 mg, 0.27 mmol) and Ruphos Pd G3 (139 mg, 0.17 mmol) were successively added to a solution of 1,8-dichloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-N-(3-methyloxetan-3-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)imidazo[1,5-a]pyridine-6-sulfonamide (1 g, 1.67 mmol) and (S)-tert-butyl 2-(methoxymethyl)piperazine-1-carboxylate (576 mg, 2.50 mmol) in 1,4-dioxane (20 mL). The reaction solution was purged with nitrogen three times and stirred at 80 °C for 2 hours under a nitrogen atmosphere. After the reaction was complete, water was added to the reaction solution, and the mixture was extracted twice with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0 - 30% ethyl acetate) to obtain the title compound (yellow solid, 720 mg, yield 54.5%). LC / MS (ESI) (m / z): 794 [M+H]+.

[0081] Step 2: (S)-1-Chloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-8-(3-(methoxymethyl)piper azin-1-yl)-N-(3-methyloxetan-3-yl)imidazo[1,5-a]pyridine-6-sulfonamide

[0082] Under an ice - water bath, trifluoroacetic acid (3 mL) was added to a solution of tert - butyl (S)-4-(1 - chloro - 3-(5-(difluoromethyl)-1,3,4 - thiadiazol - 2 - yl)-6-(N-(3 - methyloxetan - 3 - yl)-N-(2-(trimethylsilyl)ethoxymethyl)sulfamoyl)imidazo[1,5 - a]pyridin - 8 - yl)-2-(methoxymethyl)piperazine - 1 - carboxylate (720 mg, 0.91 mmol) in dichloromethane (10 mL). The reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction mixture was slowly added dropwise to a 5% aqueous sodium bicarbonate solution cooled in an ice - water bath and extracted twice with dichloromethane. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated by preparative HPLC (C18, H2O solution with 10 - 50% acetonitrile and 0.1% FA) to obtain the title compound (yellow solid, 278 mg, yield 54.4%). 1 1H NMR (400 MHz, DMSO - d6) δ 9.58 (s, 1H), 8.81 (s, 1H), 7.68 (t, J = 53.1 Hz, 1H), 6.82 (s, 1H), 4.62 (d, J = 6.0 Hz, 2H), 4.18 (d, J = 6.4 Hz, 2H), 3.41 - 3.39 (m, 1H), 3.38 - 3.35 (m, 4H), 3.28 (s, 3H), 3.20 - 3.11 (m, 2H), 3.06 - 2.97 (m, 2H), 2.77 - 2.68 (m, 1H), 1.49 (s, 3H). LC / MS (ESI) (m / z): 564 [M + H] + .

[0083] The following examples are prepared by referring to the preparation method of Example 1, starting from suitable starting materials, and synthesized according to a similar route to obtain a crude product, which is prepared by reverse-phase HPLC and freeze-dried to obtain the title compound.

[0084]

[0085]

[0086] Example 5: 1-Chloro-8-(4-(1-methoxycyclopropane-1-carbonyl)piperazin-1-yl)-N-(3-methyloxetan -3-yl)-3-(5-(trifluoromethyl)-1,3,4-thiadiazol-2-yl)imidazo[1,5-a]pyridine-6-sulfonamide

[0087]

[0088] Step 1: 1-Chloro-8-(4-(1-methoxycyclopropane-1-carbonyl)piperazin-1-yl)-N-(3-methyloxetan -3-yl)-3-(5-(trifluoromethyl)-1,3,4-thiadiazol-2-yl)-N-((2-(trimethylsilyl)ethoxy)methyl) imidazo[1,5-a]pyridine-6-sulfonamide

[0089] Under nitrogen protection, to a solution of 1,8-dichloro-N-(3-methyloxetan-3-yl)-3-(5-(trifluoromethyl)-1,3,4-thiadiazol-2-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)imidazo[1,5-a]pyridine-6-sulfonamide (80 mg, 0.13 mmol) in 1,4-dioxane (3 mL) were successively added (1-methoxycyclopropyl)(piperazin-1-yl)methanone (48 mg, 0.26 mmol), Ruphos (10 mg, 0.02 mmol), Ruphos Pd G3 (14 mg, 0.02 mmol) and cesium carbonate (127 mg, 0.36 mmol). The reaction mixture was stirred at 80 °C for 3 hours under nitrogen protection. After completion of the reaction, the reaction mixture was diluted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was separated by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0 - 100% ethyl acetate) to obtain the title compound (yellow solid, 30 mg, yield 30.2%). LC / MS (ESI) (m / z): 766 [M+H] + .

[0090] Step 2: 1-chloro-8-(4-(1-methoxycyclopropane-1-carbonyl)piperazin-1-yl)-N-(3-methyloxetane -3-yl)-3-(5-(trifluoromethyl)-1,3,4-thiadiazol-2-yl)imidazo[1,5-a]pyridine-6-sulfonamide

[0091] At 0 °C, trifluoroacetic acid (0.7 mL) was added to a solution of 1-chloro-8-(4-(1-methoxycyclopropane-1-carbonyl)piperazin-1-yl)-N-(3-methyloxetan-3-yl)-3-(5-(trifluoromethyl)-1,3,4-thiadiazol-2-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)imidazo[1,5-a]pyridine-6-sulfonamide (30 mg, 0.04 mmol) in dichloromethane (2 mL). The reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (C18, H2O solution of 30 - 95% acetonitrile and 0.1% TFA) to obtain the title compound (yellow solid, 13.7 mg, yield 55.0%). 1 HNMR (400 MHz, DMSO-d6) δ 9.59 (s, 1H), 6.92 (s, 1H), 4.64 (d, J = 6.2 Hz, 2H), 4.19 (d, J = 6.4 Hz, 2H), 4.13 - 3.63 (m, 4H), 3.27 (s, 3H), 3.16 (s, 4H), 1.50 (s, 3H), 1.06 - 1.01 (m, 2H), 0.95 - 0.90 (m, 2H). LC / MS (ESI) (m / z): 636 [M+H] + .

[0092] Example 6: 8-(4-(1-ethylcyclopropane-1-carbonyl)piperazin-1-yl)-N-(3-methyloxetane-3-yl)-3-(5-(trifluoromethyl)-1,3,4-thiadiazol-2-yl)imidazo[1,5-a]pyridine-6-sulfonamide

[0093]

[0094] Step 1: 8-(4-(1-ethylcyclopropane-1-carbonyl)piperazin-1-yl)-N-(3-methyloxetane-3-yl)- 3-(5-(trifluoromethyl)-1,3,4-thiadiazol-2-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)

[0095] imidazo[1,5-a]pyridine-6-sulfonamide

[0096] Under nitrogen protection, to a solution of 8-chloro-N-(3-methyloxetan-3-yl)-3-(5-(trifluoromethyl)-1,3,4-thiadiazol-2-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)imidazo[1,5-a]pyridine-6-sulfonamide (40 mg, 0.068 mmol) in 1,4-dioxane (3 mL) were successively added (1-ethylcyclopropyl)(piperazin-1-yl)methanone (18 mg, 0.11 mmol), Ruphos (1 mg, 0.01 mmol), Ruphos Pd G3 (6 mg, 0.005 mmol) and cesium carbonate (45 mg, 0.14 mmol). The reaction mixture was stirred at 80 °C for 3 hours under nitrogen protection. After completion of the reaction, the reaction mixture was diluted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was separated by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0 - 80% ethyl acetate) to obtain the title compound (yellow solid, 30 mg, yield 60%). LC / MS (ESI) (m / z): 730 [M+H] + .

[0097] Step 2: 8-(4-(1-ethylcyclopropane-1-carbonyl)piperazin-1-yl)-N-(3-methyloxetane-3-yl)- 3-(5-(trifluoromethyl)-1,3,4-thiadiazol-2-yl)imidazo[1,5-a]pyridine-6-sulfonamide

[0098] At 0 °C, trifluoroacetic acid (1 mL) was added to a solution of 8-(4-(1-ethylcyclopropane-1-carbonyl)piperazin-1-yl)-N-(3-methyloxetan-3-yl)-3-(5-(trifluoromethyl)-1,3,4-thiadiazol-2-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)imidazo[1,5-a]pyridine-6-sulfonamide (30 mg, 0.04 mmol) in dichloromethane (2 mL). The reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (C18, H2O solution of 10 - 50% acetonitrile and 0.1% FA) to obtain the title compound (yellow solid, 6 mg, yield 24%). 11H NMR (400 MHz, DMSO-d6) δ 9.58 (s, 1H), 8.74 (s, 1H), 8.13 (s, 1H), 6.76 (s, 1H), 4.64 (d, J = 6.2 Hz, 2H), 4.19 (d, J = 6.3 Hz, 2H), 3.87 - 3.76 (m, 4H), 3.33 - 3.32 (m, 4H), 2.08 - 1.96 (m, 2H), 1.57 - 1.52 (m, 2H), 1.50 (s, 3H), 0.91 (t, J = 7.4 Hz, 3H), 0.84 - 0.82 (m, 2H). LC / MS (ESI) (m / z): 600 [M+H] + .

[0099] The following examples were synthesized from appropriate starting materials according to a similar route to the preparation method of Example 6 to obtain a crude product, and the crude product was prepared by reverse-phase HPLC and freeze-dried to obtain the title compound. Example 10: 8-((3S,5S)-4-cyclopropyl-3,5-dimethylpiperazin-1-yl)-3-(5-(difluoromethyl)-1,

[0100]

[0101]

[0102] 3,4-thiadiazol-2-yl)-N-(3-methyloxetane-3-yl)imidazo[1,5-a]pyridine-6-sulfonamide Step 1: (2S,6S)-4-(3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-6-(N-(3-methyloxetane

[0103]

[0104] -3-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)aminosulfonyl)imidazo[1,5-a]pyridin-8- yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester Step 2: 3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-8-((3S,5S)-3,5-dimethylpiperazin-1-

[0105] Under nitrogen protection, cesium carbonate (172.7 mg, 0.53 mmol), Ruphos (16.5 mg, 0.035 mmol) and Ruphos Pd G3 (29.6 mg, 0.035 mmol) were successively added to a solution of 8-chloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-N-(3-methyloxetan-3-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)imidazo[1,5-a]pyridine-6-sulfonamide (100 mg, 0.18 mmol) and tert-butyl (2S,6S)-2,6-dimethylpiperazine-1-carboxylate (56.8 mg, 0.27 mmol) in 1,4-dioxane (2 mL). The reaction mixture was purged with nitrogen three times and stirred at 80 °C for 2 hours under nitrogen protection. After the reaction was complete, the reaction solution was diluted with ethyl acetate, washed successively with water and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0 - 35% ethyl acetate) to obtain the title compound (yellow solid, 120 mg, yield 91.4%). LC / MS (ESI) (m / z): 744 [M+H] +.

[0106] yl)-N-(3-methyloxetane-3-yl)imidazo[1,5-a]pyridine-6-sulfonamide Step 3: 8-((3S,5S)-4-cyclopropyl-3,5-dimethylpiperazin-1-yl)-3-(5-(difluoromethyl)-1,3,4-

[0107] (2S,6S)-4-(3-(5-(Difluoromethyl)-1,3,4-thiadiazol-2-yl)-6-(N-(3-methyloxetan-3-yl)-N-(2-(trimethylsilyl)ethoxymethyl)aminosulfonyl)imidazo[1,5-a]pyridin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (120 mg, 0.16 mmol), dichloromethane (2 mL), and trifluoroacetic acid (0.6 mL) were added to a reaction flask. The reaction solution was stirred at room temperature for 30 minutes. After the reaction was complete, the solution was made alkaline with sodium bicarbonate, extracted with dichloromethane, washed with saturated brine, dried, and concentrated to obtain the title compound (yellow solid, 70 mg, yield 84.5%). LC / MS (ESI) (m / z): 514 [M+H] + .

[0108] thiadiazol-2-yl)-N-(3-methyloxetane-3-yl)imidazo[1,5-a]pyridine-6-sulfonamide ​

[0109] At room temperature, sodium cyanoborohydride (7.8 mg, 0.20 mmol) and acetic acid (76.3 mg, 0.41 mmol) were added to a solution of 3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-8-((3S,5S)-3,5-dimethylpiperazin-1-yl)-N-(3-methyloxetan-3-yl)imidazo[1,5-a]pyridine-6-sulfonamide (70 mg, 0.14 mmol) and (1-ethoxycyclopropoxy)trimethylsilane (47.5 mg, 0.27 mmol) in tetrahydrofuran (1 mL). The reaction solution was stirred at 60 °C for 4 hours under nitrogen protection. After the reaction was complete, the reaction solution was diluted with ethyl acetate, washed successively with water and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated by preparative HPLC (C18, H2O solution of 10 - 50% acetonitrile and 0.1% FA) to obtain the title compound (yellow solid, 1.54 mg, yield 2.0%). 1 1H NMR (400 MHz, CDCl3) δ 9.86 (s, 1H), 7.73 (s, 1H), 7.00 (t, J = 53.6 Hz, 1H), 6.68 (s, 1H), 4.82 - 4.80 (m, 2H), 4.42 - 4.39 (m, 2H), 3.43 - 3.22 (m, 7H), 1.72 (s, 3H), 1.39 (s, 6H), 0.88 - 0.79 (m, 4H). LC / MS (ESI) (m / z): 554 [M+H] +.

[0110] Example 11: 3-(5-(Difluoromethyl)-1,3,4-thiadiazol-2-yl)-8-((3S,5S)-4-(2-hydroxyethyl)-3,5-dimethylpiperazin-1-yl)-N-(3-methyloxetan-3-yl)imidazo[1,5-a]pyridine-6-sulfonamide

[0111]

[0112] Step 1: Benzyl (3S,5S)-3,5-dimethyl-4-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)piperazine-1-carboxylate

[0113] Under nitrogen protection, potassium carbonate (331 mg, 2.40 mmol) and 2-(2-bromoethoxy)tetrahydro-2H-pyran (446 mg, 1.8 mmol) were successively added to a solution of benzyl (3S,5S)-3,5-dimethylpiperazine-1-carboxylate (300 mg, 1.20 mmol) in DMF (5 mL). The reaction solution was purged with nitrogen three times and stirred at 80 °C for 3 hours under nitrogen protection. After the reaction was complete, the reaction solution was cooled to room temperature, diluted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0 - 45% ethyl acetate) to obtain the title compound (white solid, 210 mg, yield 46%). LC / MS (ESI) m / z: 377 [M+H] + .

[0114] Step 2: (2S,6S)-2,6-Dimethyl-1-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)piperazine

[0115] At room temperature, 10% palladium on carbon (80 mg) was added to benzyl (3S,5S)-3,5-dimethyl-4-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)piperazine-1-carboxylate (210 mg, 0.55 mmol) in ethyl acetate (20 mL). The reaction mixture was purged with hydrogen three times and stirred at room temperature for 3 hours under 1 atm of hydrogen. After the reaction was complete, the reaction solution was filtered and the filtrate was concentrated under reduced pressure to obtain the title compound (colorless oil, 120 mg, yield 88%). LC / MS (ESI) (m / z): 243 [M+H] + .

[0116] Step 3: 3-(5-(Difluoromethyl)-1,3,4-thiadiazol-2-yl)-8-((3S,5S)-3,5-dimethyl-4-(2-(tetrahydro-2H-pyran-2-yl)oxy)ethyl)piperazin-1-yl)-N-(3-methyloxetan-3-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)imidazo[1,5-a]pyridine-6-sulfonamide

[0117] Under nitrogen protection, to a solution of 8-chloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-N-(3-methyloxetan-3-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)imidazo[1,5-a]pyridine-6-sulfonamide (30 mg, 0.05 mmol) in dioxane (2 mL) were successively added (2S,6S)-2,6-dimethyl-1-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)piperazine (331 mg, 0.08 mmol), cesium carbonate (45 mg, 0.18 mmol), Ruphos (5 mg, 0.01 mmol) and Ruphos Pd G3 (5 mg, 0.006 mmol). The reaction solution was purged with nitrogen three times and stirred at 80 °C for 3 hours under nitrogen protection. After the reaction was complete, the reaction solution was cooled to room temperature, diluted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0 - 45% ethyl acetate) to obtain the title compound (yellow solid, 30 mg, yield 69%). LC / MS (ESI) m / z: 772 [M+H] + .

[0118] Step 4: 3-(5-(Difluoromethyl)-1,3,4-thiadiazol-2-yl)-8-((3S,5S)-4-(2-hydroxyethyl)-3,5-dimethylpiperazin-1-yl)-N-(3-methyloxetan-3-yl)imidazo[1,5-a]pyridine-6-sulfonamide

[0119] At 0 °C, trifluoroacetic acid (0.5 mL) was added to a solution of 3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-8-((3S,5S)-3,5-dimethyl-4-(2-(tetrahydro-2H-pyran-2-yl)oxy)ethyl)piperazin-1-yl)-N-(3-methyloxetan-3-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)imidazo[1,5-a]pyridine-6-sulfonamide (30 mg, 0.03 mmol) in dichloromethane (2 mL). The mixture was stirred at room temperature for 30 minutes. The mixture was concentrated under reduced pressure, and the residue was purified by preparative HPLC to obtain the title compound (yellow solid, 2.43 mg, yield 5%).1 1H NMR (400 MHz, CD3OD) δ 9.89 (s, 1H), 7.91 (s, 1H), 7.34 (t, J = 53.5 Hz, 1H), 6.84 (d, J = 1.0 Hz, 1H), 4.82–4.77 (m, 4H), 4.35–4.31 (m, 2H), 4.23–4.05 (m, 2H), 3.98–3.92 (m, 2H), 3.86–3.73 (m, 2H), 3.67–3.61 (m, 1H), 3.54–3.46 (m, 1H), 1.65 (s, 3H), 1.64–1.53 (m, 6H). LC / MS (ESI) (m / z): 558 [M+H] + .

[0120] Example 12: 8-((2S,6S)-2,6-Dimethylmorpholino)-3-(5-(hydroxymethyl)-1,3,4-thiadiazol-2-yl)-N-(3-methyloxetan-3-yl)imidazo[1,5-a]pyridine-6-sulfonamide

[0121]

[0122] Step 1: Methyl (5-(8-((2S,6S)-2,6-dimethylmorpholino)-6-(N-(3-methyloxetan-3-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)sulfamoyl)imidazo[1,5-a]pyridin-3-yl)-1,3,4-thiadiazol-2-yl)acetate

[0123] Under nitrogen protection, cesium carbonate (45 mg, 0.18 mmol), Ruphos (5 mg, 0.01 mmol) and Ruphos Pd G3 (5 mg, 0.006 mmol) were successively added to a solution of methyl (5-(8-chloro-6-(N-(3-methyloxetan-3-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)sulfamoyl)imidazo[1,5-a]pyridin-3-yl)-1,3,4-thiadiazol-2-yl)acetate (35 mg, 0.06 mmol) and 2S,6S-dimethylmorpholine (9 mg, 0.07 mmol) in 1,4-dioxane (2 mL). The reaction solution was purged with nitrogen three times and stirred at 80 °C for 5 hours under nitrogen protection. After the reaction was complete, the reaction solution was cooled to room temperature, diluted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0 - 45% ethyl acetate) to obtain the title compound (white solid, 30 mg, yield 74%). LC / MS (ESI) m / z: 667 [M+H] + .

[0124] Step 2: Methyl (5-(8-((2S,6S)-2,6-dimethylmorpholino)-6-(N-(3-methyloxetan-3-yl)sulfamoyl)imidazo[1,5-a]pyridin-3-yl)-1,3,4-thiadiazol-2-yl)acetate

[0125] At 0 °C, trifluoroacetic acid (1 mL) was successively added to a solution of methyl (5-(8-((2S,6S)-2,6-dimethylmorpholino)-6-(N-(3-methyloxetan-3-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)sulfamoyl)imidazo[1,5-a]pyridin-3-yl)-1,3,4-thiadiazol-2-yl)acetate (13 mg, 0.01 mmol) in dichloromethane (3 mL). The reaction mixture was stirred at 0 °C for 1 h. After completion of the reaction, water was added, and the mixture was extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0 - 80% ethyl acetate) to give the title compound (white solid, 10 mg, yield 95%). LC / MS (ESI) (m / z): 537 [M + H] + .

[0126] Step 3: 8-((2S,6S)-2,6-Dimethylmorpholino)-3-(5-(hydroxymethyl)-1,3,4-thiadiazol-2-yl)-N-(3-methyloxetan-3-yl)imidazo[1,5-a]pyridine-6-sulfonamide

[0127] At 0 °C, tetrahydrofuran (1 mL), water (1 mL) and lithium hydroxide (1 mg, 0.03 mmol) were added to a solution of methyl (5-(8-((2S,6S)-2,6-dimethylmorpholino)-6-(N-(3-methyloxetan-3-yl)sulfamoyl)imidazo[1,5-a]pyridin-3-yl)-1,3,4-thiadiazol-2-yl)acetate (10 mg, 0.018 mmol) in methanol (1 mL). The mixture was stirred at room temperature for 60 min. The mixture was concentrated under reduced pressure, and the residue was purified by preparative HPLC to give the title compound (white solid, 0.41 mg, yield 5%). 1 1H NMR (400 MHz, CD3OD) δ 9.82 (s, 1H), 7.77 (s, 1H), 6.68 (s, 1H), 5.18–5.16 (m, 2H), 4.32–4.28 (m, 4H), 3.49–3.47 (m, 4H), 3.13–3.12 (m, 2H), 1.63 (s, 3H), 1.41–1.39 (m, 6H). LC / MS (ESI) (m / z): 495 [M + H] + .

[0128] Example 13: 3-(5-(Hydroxymethyl)-1,3,4-thiadiazol-2-yl)-8-(3-(methoxymethyl)piperazin-1-yl)-N-(3-methyloxetan-3-yl)imidazo[1,5-a]pyridine-6-sulfonamide

[0129]

[0130] Step 1: Methyl (5-(8-(tert-Butyl 2-(methoxymethyl)piperazine-1-carboxylate)-6-(N-(3-methyloxetan-3-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)sulfamoyl)imidazo[1,5-a]pyridin-3-yl)-1,3,4-thiadiazol-2-yl)acetate

[0131] Under nitrogen protection, cesium carbonate (45 mg, 0.18 mmol), Ruphos (5 mg, 0.01 mmol) and Ruphos Pd G3 (5 mg, 0.006 mmol) were successively added to a solution of methyl (5-(8-chloro-6-(N-(3-methyloxetan-3-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)sulfamoyl)imidazo[1,5-a]pyridin-3-yl)-1,3,4-thiadiazol-2-yl)acetate (35 mg, 0.06 mmol) and tert-butyl 2-(methoxymethyl)piperazine-1-carboxylate (16 mg, 0.07 mmol) in 1,4-dioxane (2 mL). The reaction mixture was purged with nitrogen three times and stirred at 80 °C for 2 h under nitrogen protection. After completion of the reaction, the reaction mixture was cooled to room temperature, diluted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0 - 45% ethyl acetate) to give the title compound (white solid, 30 mg, yield 64%). LC / MS (ESI) m / z: 782 [M+H] + .

[0132] Step 2: Methyl (5-(8-(3-(methoxymethyl)piperazin-1-yl)-6-(N-(3-methyloxetan-3-yl)sulfamoyl)imidazo[1,5-a]pyridin-3-yl)-1,3,4-thiadiazol-2-yl)acetate

[0133] At 0 °C, trifluoroacetic acid (1 mL) was successively added to a solution of methyl (5-(8-(2-(methoxymethyl)piperazine-1-carboxylate tert-butyl)-6-(N-(3-methyloxetan-3-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)sulfamoyl)imidazo[1,5-a]pyridin-3-yl)-1,3,4-thiadiazol-2-yl)acetate (30 mg, 0.03 mmol) in dichloromethane (3 mL). The reaction mixture was stirred at 0 °C for 1 hour. After completion of the reaction, water was added and the mixture was extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0 - 80% ethyl acetate) to obtain the title compound (white solid, 20 mg, yield 95%). LC / MS (ESI) (m / z): 552 [M+H] + .

[0134] Step 3: 3-(5-(Hydroxymethyl)-1,3,4-thiadiazol-2-yl)-8-(3-(methoxymethyl)piperazin-1-yl)-N-(3-methyloxetan-3-yl)imidazo[1,5-a]pyridine-6-sulfonamide

[0135] At 0 °C, tetrahydrofuran (1 mL), water (1 mL) and lithium hydroxide (2 mg, 0.09 mmol) were added to a solution of methyl (5-(8-(3-(methoxymethyl)piperazin-1-yl)-6-(N-(3-methyloxetan-3-yl)sulfamoyl)imidazo[1,5-a]pyridin-3-yl)-1,3,4-thiadiazol-2-yl)acetate (20 mg, 0.03 mmol) in methanol (1 mL). The mixture was stirred at room temperature for 60 minutes. The mixture was concentrated under reduced pressure and the residue was purified by preparative HPLC to obtain the title compound (yellow solid, 0.93 mg, yield 5%). 1 H NMR (400 MHz, CD3OD) δ 9.90 (s, 1H), 7.89 (s, 1H), 6.78 (s, 1H), 4.80–4.78 (m, 2H), 4.34–4.30 (m, 2H), 3.94–3.86 (m, 2H), 3.85–3.74 (m, 3H), 3.70–3.60 (m, 2H), 3.56–3.51 (m, 2H), 3.48 (s, 3H), 3.23–3.15 (m, 2H), 1.65 (s, 3H). LC / MS (ESI) (m / z): 510 [M+H] + .

[0136] Effect Example

[0137] Experimental method:

[0138] 1. Biochemical Assay

[0139] 1) Use Echo 550 (LABCYTE, #550) to add 20 nL of compounds with different concentrations to a 384-well plate (the final concentrations of the compounds are: 1000 nM, 300 nM, 100 nM, 30 nM, 10 nM, 3 nM, 1 nM, 0.3 nM, 0.1 nM, 0.03 nM, 0 nM);

[0140] 2) Prepare reaction buffer I: 50 mM Tris (2-amino-2-hydroxymethyl-1,3-propanediol, Sigma, #T1503) at pH 7.4, 0.015% Triton X-100 (4-(1,1,3,3-tetramethylbutyl)phenyl-polyethylene glycol, Sigma, #T8787), 0.1% BSA (bovine serum albumin, Sigma, #A1933), 5 mM KCl, 66.2 μM TFMU-ADPr (fluorescein-4-trifluoromethylumbelliferyl-ADP-ribose);

[0141] 3) Add reaction buffer I to the 384-well plate at 10 μL / well;

[0142] 4) Prepare reaction buffer II: 50 mM Tris at pH 7.4, 0.015% Triton X-100, 0.1% BSA, 5 mM KCl, 4 nM PARG (poly(ADP-ribose) glycohydrolase);

[0143] 5) Add reaction buffer II to the 384-well plate at 10 μL / well to initiate the reaction;

[0144] 6) Incubate at room temperature for 3 min for the reaction;

[0145] 7) Place the 384-well plate in a microplate reader, shake for 5 s, and then perform signal detection;

[0146] 8) Use GraphPad software to analyze the data and determine the IC50.

[0147] 2. Cell Viability Assay

[0148] 1) Seed cells in a 384-well cell culture plate. HCC1806 (ATCC cell bank, 100 cells / well) and MDA-MB-231 (ATCC cell bank, 120 cells / well) are placed in a CO2 (5%) incubator at 37 °C and incubated for 24 h to allow the cells to adhere completely;

[0149] 2) Add the culture medium (100 μL / well) containing the test compound at different concentrations (final concentration of the compound: 30000 nM, 10000 nM, 3000 nM, 1000 nM, 300 nM, 100 nM, 30 nM, 10 nM, 3 nM, 1 nM, 0 nM) to the cells and incubate them in a CO2 (5%) incubator at 37 °C for 7 days;

[0150] 3) Place the culture plate containing the cells at room temperature for 30 min to equilibrate the plate and its contents to room temperature;

[0151] 4) Add an equal volume of CellTiter-Glo (Promega: V9101) reagent to each well of the cells, mix the contents on an orbital shaker for 2 minutes to induce cell lysis;

[0152] 5) Incubate the plate at room temperature for 10 minutes. After the signal value stabilizes, use a microplate reader (Thermo: Varioskan LUX) to detect and record the luminescence value;

[0153] 6) Use GraphPad software to analyze the data and determine the IC 50 .

[0154] Data list:

[0155] NT = No detection;

[0156]

[0157]

[0158] Tested, the example compound of this application has a strong inhibitory effect on the proliferation of HCC1806 cells and a weak inhibitory effect on the proliferation of MDA-MB-231 cells, indicating that the example compound of this application has good selectivity for the inhibitory effect on the proliferation of the two types of cells.

Claims

1. Any one of the following compounds, or a pharmaceutically acceptable salt or isotope compound thereof:

2. A pharmaceutical composition comprising a substance Z and a pharmaceutical excipient, wherein the substance Z is any compound according to claim 1, or a pharmaceutically acceptable salt or isotope compound thereof.

3. Use of a substance Z in the preparation of a PARG inhibitor and a drug for treating and / or preventing PARG-related diseases, wherein the substance Z is any compound as claimed in claim 1, or a pharmaceutically acceptable salt or isotope compound thereof.

4. The use according to claim 3, characterized in that The PARG-related disease is breast cancer.

5. The use according to claim 4, characterized in that The PARG-related disease is triple-negative breast cancer.

6. Use of a substance Z in the preparation of a drug for treating and / or preventing breast cancer, wherein the substance Z is any compound as claimed in claim 1, or a pharmaceutically acceptable salt or isotope compound thereof.

7. The use according to claim 6, characterized in that The breast cancer is triple-negative breast cancer.

8. A substance Z for treating and / or preventing PARG-related diseases, wherein the substance Z is any compound according to claim 1, or a pharmaceutically acceptable salt or isotope compound thereof.

9. The substance Z according to claim 8, characterized in that The PARG-related disease is breast cancer, such as triple-negative breast cancer.

10. A method for treating and / or preventing PARG-related diseases, comprising administering an effective amount of substance Z to a patient, wherein the substance Z is any compound as described in claim 1, a pharmaceutically acceptable salt or isotope compound thereof; the PARG-related disease may be breast cancer, such as triple-negative breast cancer.

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