Thiazolopyrimidone compound as well as composition and application thereof

Abstract

The invention provides a thiazolopyrimidone compound as well as a composition and application thereof, and the thiazolopyrimidone compound as well as a derivative and salt thereof provided by the invention have the effects of improving sleep and resisting depression, and can be used as a GABAA receptor positive allosteric modulator. The structural formula of the thiazolopyrimidone compound is as shown in formula (I),

Description

Technical Field

[0001] The present invention relates to the technical field of medicinal chemistry, and in particular to a thiazolopyrimidinone compound, a composition thereof, and uses thereof. Background Art

[0002] GABA receptor type A A GABA receptors are the most important inhibitory neurotransmitter receptors in the central nervous system. Their functional regulation is closely related to various neuropsychiatric disorders, including anxiety, epilepsy, sleep disorders, depression, and drug addiction. Positive allosteric modulators (PAMs), which enhance the binding efficiency of γ-aminobutyric acid (GABA) to receptors, have become an important direction in the development of therapeutic drugs for neuropsychiatric diseases.

[0003] GABA currently used in clinical practice A Benzodiazepine receptor modulators diazepam), non-benzodiazepines However, existing drugs generally have the following defects: (1) insufficient subtype selectivity leads to dose-dependent side effects, including excessive sedation, cognitive impairment, and respiratory depression; (2) long-term use is prone to tolerance and drug dependence; (3) there is a lack of synergistic therapeutic effect on complex disease symptoms (such as depression with sleep disorders). Summary of the Invention

[0004] In order to solve the above technical problems, the present invention develops a thiazolylpyrimidinone compound with a novel substitution pattern, high subtype selectivity and multi-target therapeutic effect, which promotes GABA A Clinical applications of receptor modulators.

[0005] One object of the present invention is to provide a thiazolopyrimidinone compound;

[0006] Another object of the present invention is to provide a pharmaceutical composition containing a thiazolopyrimidone compound;

[0007] Another object of the present invention is to provide uses of the thiazolopyrimidone compounds.

[0008] To achieve the above objectives, in one aspect, the present invention provides a thiazolopyrimidinone compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the structural formula of the thiazolopyrimidinone compound is shown in formula (I):

[0009]

[0010] wherein R1 is selected from substituted or unsubstituted aryl, heteroaryl or saturated heterocyclic group; when substituted, the aryl, heteroaryl or saturated heterocyclic group is substituted by a substituent selected from halogen, C1-C6 alkyl, C1-C6 alkoxy, hydroxyl, trifluoromethyl or trifluoromethoxy; the heteroaryl or saturated heterocyclic group contains 1, 2, 3 or 4 heteroatoms selected from N, O or S;

[0011] R2 is selected from substituted or unsubstituted C1-C6 alkyl, C3-C6 cycloalkyl, 6-10 membered aryl, 5-10 membered heteroaryl; when substituted, the alkyl, cycloalkyl, aryl or heteroaryl is substituted with a substituent selected from halogen, C1-C6 alkyl, C1-C6 alkoxy, cyano or nitro; the heteroaryl contains 1 or 2 heteroatoms selected from N, O or S;

[0012] R3 is selected from substituted or unsubstituted 6-10 membered aryl or 5-10 membered heteroaryl; when substituted, the aryl or heteroaryl is substituted by a substituent selected from halogen, C1-C6 alkyl, C1-C6 alkoxy, cyano or nitro; the heteroaryl contains 1, 2, 3 or 4 heteroatoms selected from N, O or S.

[0013] According to some specific embodiments of the present invention,

[0014] R1 is selected from substituted or unsubstituted phenyl, substituted or unsubstituted 5-6 membered heteroaryl, unsubstituted 4-6 membered saturated heterocyclic group; when substituted, the phenyl or heteroaryl group is substituted with a substituent selected from fluorine, chlorine, bromine, C1-C6 alkyl, C1-C6 alkoxy, hydroxyl, trifluoromethyl or trifluoromethoxy;

[0015] R2 is selected from C1-C6 alkyl, C3-C6 cycloalkyl, substituted or unsubstituted 6-10 membered aryl, 5-6 membered heteroaryl; when substituted, the aryl or heteroaryl is substituted with a substituent selected from fluorine, chlorine, bromine, C1-C6 alkyl, C1-C6 alkoxy, cyano or nitro;

[0016] R3 is selected from substituted or unsubstituted 6-10 membered aryl or 5-10 membered heteroaryl; when substituted, the aryl or heteroaryl is substituted by a substituent selected from fluorine, chlorine, bromine, C1-C6 alkyl, C1-C6 alkoxy, cyano or nitro.

[0017] According to some specific embodiments of the present invention,

[0018] R1is selected from a 6-membered saturated heterocyclyl, substituted or unsubstituted phenyl or 5-6 membered heteroaryl; when substituted, the phenyl or heteroaryl is substituted with a substituent selected from fluorine, chlorine, bromine, methyl, ethene, n-propyl, iso-propyl, tert-butyl, methoxy, ethoxy, hydroxy, trifluoromethyl or trifluoromethoxy; the heteroaryl or saturated heterocyclyl contains 1, 2 or 3 heteroatoms selected from N, O or S;

[0019] R2is selected from C1-C4 alkyl, C3-C6 cycloalkyl, substituted or unsubstituted phenyl or 6-membered heteroaryl; when substituted, the phenyl or heteroaryl is substituted with a substituent selected from fluorine, chlorine, bromine, C1-C6 alkyl or C1-C6 alkoxy; the heteroaryl contains 1, 2, 3 heteroatoms selected from N, O or S;

[0020] R3is selected from substituted or unsubstituted phenyl or 6-membered heteroaryl; when substituted, the phenyl or heteroaryl is substituted with a substituent selected from fluorine, chlorine, bromine, C1-C6 alkyl, C1-C6 alkoxy, or nitro; the heteroaryl contains 1, 2 or 3 heteroatoms selected from N, O or S.

[0021] According to some embodiments of the application, wherein,

[0022] In R1, the heteroaryl contains 1 or 2 heteroatoms selected from N or S;

[0023] In R2, the heteroaryl contains 1 or 2 N atoms;

[0024] In R3, the heteroaryl contains 1 or 2 N atoms.

[0025] According to some embodiments of the application, wherein,

[0026] R1is selected from a 6-membered saturated heterocyclyl, substituted or unsubstituted phenyl or 5-6 membered heteroaryl; when substituted, the phenyl or heteroaryl is substituted with a substituent selected from fluorine, chlorine, bromine, methyl, methoxy, hydroxy or trifluoromethyl;

[0027] R2is selected from methyl, ethyl, iso-propyl, cyclopropyl, substituted or unsubstituted phenyl, pyrimidinyl or pyrimidinyl; when substituted, the phenyl, pyrimidinyl or pyrimidinyl is substituted with a substituent selected from fluorine, chlorine or methyl;

[0028] R3is selected from substituted or unsubstituted phenyl, pyridinyl or pyrimidinyl; when substituted, the phenyl, pyridinyl or pyrimidinyl is substituted with a substituent selected from fluorine, chlorine, methyl or nitro.

[0029] According to some embodiments of the application, wherein,

[0030] R1 is selected from morpholinyl, thienyl, pyrimidinyl, methoxypyrimidinyl, hydroxyphenyl, fluorophenyl, methoxyphenyl, pyridyl, imidazolyl;

[0031] R2 is selected from methyl, ethyl, cyclopropyl or phenyl;

[0032] R3 is selected from pyridyl substituted by methyl, substituted or unsubstituted phenyl; when substituted, the phenyl is substituted by 1 or 2 substituents selected from methyl, ethyl, fluorine or chlorine.

[0033] According to some specific embodiments of the present invention,

[0034] R1 is selected from thienyl or pyrimidinyl substituted by methoxy;

[0035] R2 is selected from methyl, ethyl, cyclopropyl or unsubstituted phenyl;

[0036] R3 is selected from phenyl groups substituted by methyl, ethyl, fluorine or chlorine, and the number of substituents of the phenyl group is 1 or 2.

[0037] According to some specific embodiments of the present invention, the thiazolopyrimidinone compound is selected from one of the following structures:

[0038]

[0039]

[0040] According to some specific embodiments of the present invention, the thiazolopyrimidinone compound is selected from one of the following structures:

[0041]

[0042] According to some specific embodiments of the present invention, the pharmaceutically acceptable salt is hydrochloride, sulfate, methanesulfonate or maleate.

[0043] On the other hand, the present invention also provides a pharmaceutical composition, which contains the above-mentioned thiazolopyrimidone compound, its stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0044] According to some specific embodiments of the present invention, the pharmaceutical composition is in a gastrointestinal dosage form or an injectable dosage form.

[0045] According to some specific embodiments of the present invention, the gastrointestinal administration dosage form is selected from powders, tablets, granules, capsules, solutions, emulsions or suspensions.

[0046] According to some specific embodiments of the present invention, the injection dosage form is selected from intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection or intracavitary injection dosage form.

[0047] In another aspect, the present invention also provides use of the thiazolopyrimidone compound, its stereoisomer or pharmaceutically acceptable salt thereof, or the pharmaceutical composition of the present invention in the preparation of a drug for improving sleep or antidepressant effects.

[0048] According to some specific embodiments of the present invention, the thiazolopyrimidinone compound, its stereoisomer or its pharmaceutically acceptable salt is used as GABA A Receptor modulators.

[0049] According to some specific embodiments of the present invention, the depression includes mild, moderate and severe depression.

[0050] The present invention defines the terms used herein as follows:

[0051] The term "GABA A "Receptor" refers to the gamma-aminobutyric acid type A receptor.

[0052] The term "EC 50 ” refers to the half-maximal effective concentration, which refers to the drug concentration value that can produce half the maximum effect.

[0053] The term "PAM" refers to positive allosteric modulator.

[0054] The term "α4β3δ" refers to GABA A One of many subtypes of receptors.

[0055] The term "relative current enhancement percentage (%)" refers to the inward current (I C ) minus the current induced by the same concentration of GABA alone (I G ), the difference between the obtained value and I G The ratio of (I C -I G ) / I G ×100%.

[0056] "Aryl" refers to a substituted or unsubstituted aromatic hydrocarbon group having a single ring or fused rings. The aryl ring may be fused to a saturated or unsaturated carbocyclic or heterocyclic ring, wherein the ring connecting to the parent structure is the aryl ring, non-limiting examples of which include benzene rings and naphthalene rings. "Aryl" can be monovalent, divalent, trivalent, or tetravalent. When divalent, trivalent, or tetravalent, the point of attachment is on the aryl ring.

[0057] "Heteroaryl" refers to a substituted or unsubstituted aromatic hydrocarbon group containing 1 to 4 heteroatoms or groups containing heteroatoms (including but not limited to N, O or S(=O)n, where n is 0, 1, or 2), and the number of ring atoms in the heteroaromatic ring includes but is not limited to 5 to 10 or 5 to 6. Non-limiting examples of heteroaryl groups include but are not limited to pyridyl, furyl, thienyl, pyridyl, pyranyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, benzopyrazole, benzimidazole, benzopyridine, pyrrolopyridine, and the like. Heteroaryl groups can be monovalent, divalent, trivalent, or tetravalent. When divalent, trivalent, or tetravalent, the site of attachment is on the heteroaryl ring.

[0058] In summary, the present invention provides a thiazolopyrimidone compound, a composition thereof, and uses thereof. The technical solution of the present invention has the following advantages:

[0059] In vitro electrophysiological activity experiments have shown that the thiazolylpyrimidone compounds and their derivatives and salts provided by the present invention have certain GABA A R positive allosteric activity. Compared with allopregnanolone and positive compound DS-2, using two-electrode voltage clamp technique, the thiazolopyrimidinone compounds of the present invention have a positive effect on GABA at a concentration of 10 μM (micromolar). A The receptor exhibits significant positive allosteric activity and is equal to or close to the maximum positive allosteric activity, and the activity-concentration dose-effect relationship is obvious, which is better than the clinical drug allopregnanolone and the positive compound DS-2. It is expected to become a new type of GABA with novel structure, strong activity and high selectivity. A Receptor positive allosteric modulator. BRIEF DESCRIPTION OF THE DRAWINGS

[0060] Figure 1 The dose-effect curves of the representative compounds of the present invention (WH20240720, WH20240413, LYZ-20240630), the positive drug (allopregnanolone) and the positive compound DS-2 in African clawed frog oocytes expressing α4β3δ.

[0061] Figure 2 To evaluate the activity of compound WH20240720 in a sleep deprivation mouse model.

[0062] Figure 3 This is an activity evaluation of compound WH20240720 in improving depressive-like behavior in mice, where Figure A shows a single tail suspension test after multiple administrations, and Figure B shows a single tail suspension test after a single administration. DETAILED DESCRIPTION

[0063] In order to have a clearer understanding of the technical features, objectives and beneficial effects of the present invention, the technical solution of the present invention is now described in detail below, but it should not be understood as limiting the scope of implementation of the present invention.

[0064] This section provides a general description of the materials and experimental methods used in the experiments of the present invention. Although many of the materials and procedures used to achieve the purposes of the present invention are well known in the art, the present invention is described herein in as much detail as possible. It will be understood by those skilled in the art that, unless otherwise specified, the materials and procedures used in the present invention are well known in the art.

[0065] The materials, reagents and instruments used in the examples of the present invention are as follows:

[0066] Material:

[0067] Xenopus oocytes;

[0068] Female C57BL6 / N mice (18–25 g, Department of Animal Science, Peking University Health Science Center);

[0069] Reagents:

[0070] ND96 extracellular solution: 96 mM NaCl, 2 mM KCl, 1.8 mM CaCl2, 1 mM MgCl2, 5 mM HEPES buffer (pH = 7.4);

[0071] OR2 solution: 82.5 mM NaCl, 2 mM KCl, 1 mM MgCl2, 5 mM HEPES buffer (pH = 7.4);

[0072] Glass electrode perfusion solution: 3M KCl.

[0073] Reagents:

[0074] Solvents such as petroleum ether (60–90 °C), ethyl acetate, dichloromethane, methanol, ethanol, tetrahydrofuran, acetonitrile, toluene, and diethyl ether were all chemically pure (AR) and purchased from Beijing Tongguang Fine Chemical Company;

[0075] Thin-layer chromatography silica gel plates were GF254, purchased from Shanghai Haohong Biopharmaceutical Technology Co., Ltd.;

[0076] Column chromatography silica gel was 200-300 mesh and was purchased from Qingdao Ocean Chemical Plant.

[0077] Other experimental reagents and raw materials were of chemically pure (AR) and purchased from Haihaohong Biopharmaceutical Technology Co., Ltd., Shanghai Bid Pharmaceutical Technology Co., Ltd., Beijing Tongguang Fine Chemical Co., Ltd., etc.

[0078] instrument:

[0079] GeneClamp 500B amplifier (Axon Instruments, Union City, CA, USA);

[0080] BIOPAC multi-lead physiological signal recording amplifier (BIOPAC, USA)

[0081] Origin 2020b (OriginLab, USA);

[0082] Excel 2016 (Microsoft, USA);

[0083] GraphPad Prism 9.1.1 (GraphPad Software, USA);

[0084] Electrode pulling apparatus (Model PC-10 puller, Narishige, Japan);

[0085] Polishing machine (MF-830, Narishige, Japan);

[0086] Constant temperature water bath (JULABO, Germany).

[0087] Example 1: LWM-A24

[0088] This example is used to illustrate the preparation and identification of the compound 6-(2-chloro-6-methylphenyl)-2-(2-thienyl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0089] Target compound structure:

[0090] English name:

[0091] 6-(2-Chloro-6-methylphenyl)-5-phenyl-2-(thiophen-2-yl)thiazolo[4,5-d]pyrimidin-7(6H)-one

[0092]

[0093] The starting material is 2-chloro-6-methylaniline, which undergoes a nucleophilic substitution reaction with chloroacetyl chloride to obtain the corresponding intermediate, which is then reacted with dipotassium cyanodithiocarbamate and iodomethane to obtain the corresponding intermediate. The intermediate reacts with trimethyl orthobenzoate to obtain a key intermediate, which reacts with thiophene-2-boronic acid in the presence of cuprous thiophene-2-carboxylate and catalyzed by Pd(PPh3)4 in a Liebeskind– The coupling reaction gave compound LWM-A24.

[0094] The specific steps are as follows:

[0095] (1) The starting material 2-chloro-6-methylaniline (14 g, 100 mmol, 1.0 equiv) was dissolved in 80 mL of saturated AcONa solution and 100 mL of AcOH solution. Chloroacetyl chloride (16.8 g, 150 mmol, 1.5 equiv) was added dropwise to the solution at room temperature, followed by stirring at room temperature for about 3 hours. After the reaction was completed, 300 mL of water was added to the system to precipitate the product. The product was filtered and the solid was washed three times with water to obtain the intermediate 2-chloro-N-(2-chloro-6-methylphenyl)acetamide.

[0096] (2) The intermediate 2-chloro-N-(2-chloro-6-methylphenyl)acetamide (10.9 g, 50 mmol, 1.0 equiv) and dipotassium cyanodithiocarbamate (9.7 g, 50 mmol, 1.0 equiv) were dissolved in 200 mL of a 1 / 1 acetone-water mixture. 25 mL of a 2 M NaOH aqueous solution was added. The reaction solution was heated to 80°C and stirred for 1 hour. After the reaction solution was cooled, iodomethane (7.1 g, 50 mmol, 1.0 equiv) was added at room temperature. The mixture was stirred at room temperature for 30 minutes. Water was added to the system to precipitate the product, which was filtered and washed with water to obtain the intermediate 4-amino-N-(2-chloro-6-methylphenyl)-2-methylmercaptothiazole-5-carboxamide.

[0097] (3) The intermediate 4-amino-N-(2-chloro-6-methylphenyl)-2-methylmercaptothiazole-5-carboxamide (783 mg, 2.5 mmol, 1.0 equiv) was dissolved in 6 mL of trimethyl orthobenzoate, and 0.5 mL of acetic acid was added. The reaction was heated to 120°C and stirred for 2 days. After the reaction was completed, EA was added to the reaction solution, and the organic phase was extracted three times with saturated NaHCO3. The organic phase was dried and concentrated, and then separated by silica gel column chromatography (dry column loading, PE / EA = 10 / 1 to 3 / 1) to obtain the key intermediate 6-(2-chloro-6-methylphenyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0098] (4) The intermediate 6-(2-chloro-6-methylphenyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (399 mg, 1 mmol, 1 equiv) was dissolved in THF, and thiophene-2-boronic acid (256 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were added in sequence. The reaction system was replaced with argon 8 times, heated to 100°C, and stirred for 48 hours. After the reaction was completed, the excess metal reagent was filtered and removed with celite. The organic phase was concentrated, 70 mL of DCM was added, and the product was extracted 3 times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1, volume ratio, the same below) to obtain a crude product. The crude product was recrystallized from ethanol to obtain a light green solid LWM-A24.

[0099] Through the above preparation process, 261 mg of light green solid was obtained, with a yield of 60%.

[0100] 1 H NMR (400MHz, DMSO-d6) δ8.08(d,J=3.7Hz,1H),8.01(d,J=5.0Hz,1H),7.47–7.41(m,2H),7.41–7.34(m,2H),7.30(m,J=12.3,5H),2.17(s,3H). 13 C NMR(101MHz,DMSO-d6)δ167.6,165.6,158.6,156.0,138.5,135.3,133.8,133.6,1 33.1,131.8,131.1,130.8,130.4,129.7,129.2,127.9,127.8,127.5,114.9,18.0.

[0101] C 22 H 14 ClN3OS2 high resolution mass spectrum: 436.03349.

[0102] Example 2: LWM-A26

[0103] This example is used to illustrate the preparation and identification of the compound 6-(2-chloro-6-methylphenyl)-2-(2-furyl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0104] Target compound structure:

[0105] English name:

[0106] 6-(2-Chloro-6-methylphenyl)-2-(furan-2-yl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one

[0107] The specific steps are as follows:

[0108] Following the preparation route, referring to compound LWM-A24, the key intermediate 6-(2-chloro-6-methylphenyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (399 mg, 1 mmol, 1 equiv), 2-furanboronic acid (224 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed in a 35 mL sealed tube and stirred at 100°C under argon for 48 hours. After completion of the reaction, the excess metal reagent was filtered through Celite to remove impurities. The organic phase was concentrated, 70 mL of DCM was added, and the mixture was extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to obtain LWM-A26 as a light green solid.

[0109] Through the above preparation process, 261 mg of light green solid was obtained, with a yield of 66%.

[0110] 1 H NMR(400MHz, DMSO-d6)δ8.13(d,J=1.1Hz,1H),7.55(d,J=3.5Hz,1H),7.47–7.42(m, 2H),7.40–7.35(m,2H),7.35–7.24(m,4H),6.88(dd,J=3.5,1.7Hz,1H),2.17(s,3H). 13 C NMR(101MHz,DMSO-d6)δ166.43,163.48,159.12,156.64,148.06,147.75,139.03,134.29,134 .07,132.28,131.26,130.88,130.21,128.40,128.32,127.95,114.98,114.74,114.17,18.49.

[0111] C 22 H 14 ClN3O2S high resolution mass spectrum: 420.05619.

[0112] Example 3: LWM-A13

[0113] This example is used to illustrate the preparation and identification of compound 6-(2-chloro-6-methylphenyl)-2-(4-hydroxyphenyl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0114] Structure of the target compound:

[0115] English name:

[0116] 6-(2-Chloro-6-methylphenyl)-2-(4-hydroxyphenyl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one

[0117] The specific steps are as follows:

[0118] The preparation route refers to compound LWM-A24. The key intermediate 6-(2-chloro-6-methylphenyl)-2-methylthio-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (399 mg, 1 mmol, 1 equiv), 4-hydroxyphenylboronic acid (274 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), Pd(PPh3)4(58 mg, 0.05 mmol, 0.05 equiv) were sequentially placed in a 35 mL sealed tube, and stirred at 100°C under argon protection for 8 hours. After the reaction was completed, the excess metal reagent was removed by filtration with diatomite. After the organic phase was concentrated, 70 mL of DCM was added, and extracted with 10% NaHCO3three times. After the organic phase was concentrated, it was purified by silica gel column chromatography (PE / EA = 10 / 1 ~ 3 / 1) to obtain the crude product. The crude product was recrystallized with ethanol to obtain white solid LWM-A13.

[0119] Through the above preparation process, 276 mg of white solid was obtained, with a yield of 62%.

[0120] 1 H NMR (400 MHz, DMSO-d6) δ 10.52 (s, 1H), 8.08 - 8.01 (m, 2H), 7.43 (dd, J = 8.3, 6.8 Hz, 2H), 7.41 - 7.35 (m, 2H), 7.31 (dd, J = 12.9, 5.1 Hz, 3H), 7.29 - 7.25 (m, 1H), 7.02 - 6.95 (m, 2H), 2.17 (s, 3H). 13C NMR(101MHz,DMSO-d6)δ174.56,166.07,161.81,158.29,156.10,138.53,133.94,133.72,131 .85,130.72,130.32,129.71,129.39,127.91,127.82,127.47,123.16,116.34,114.45,18.02.

[0121] C 24 H 16 ClN3O2S high resolution mass spectrum: 446.07209.

[0122] Example 4: LWM-A27

[0123] This example is used to illustrate the preparation and identification of the compound 6-(2-chloro-6-methylphenyl)-2-(4-methoxyphenyl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0124] Target compound structure:

[0125] English name:

[0126] 6-(2-Chloro-6-methylphenyl)-2-(4-methoxyphenyl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one

[0127] The specific steps are as follows:

[0128] The preparation route was similar to compound LWM-A24. The key intermediate 6-(2-chloro-6-methylphenyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (399 mg, 1 mmol, 1 equiv), p-methoxyphenylboronic acid (304 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed sequentially in a 35 mL sealed tube and stirred at 100°C under argon for 8 hours. After completion of the reaction, the excess metal reagent was filtered through Celite to remove impurities. The organic phase was concentrated, added with 70 mL of DCM, and extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to obtain LWM-A27 as a white solid.

[0129] After the above preparation process, 266 mg of white solid was obtained, with a yield of 58%.

[0130] 1 H NMR (400MHz, DMSO-d6) δ8.16–8.09(m,2H),7.46–7.42(m,2H),7.39–7.35(m,2H),7.35–7.25(m,4H),7.20–7.14(m,2H),3.89(s,3H),2.17(s,3H). 13 C(101MHz,DMSO-d6)δ174.13,166.00,162.86,158.36,156.11,138.52,133.89,133.68,131.82, 130.73,130.34,129.71,129.18,127.90,127.82,127.46,124.62,115.02,114.84,55.67,17.99.

[0131] C 25 H 18 ClN3O2S high resolution mass spectrum: 460.08774.

[0132] Example 5: LWM-A16

[0133] This example is used to illustrate the preparation and identification of the compound 6-(2-chloro-6-methylphenyl)-2-(4-fluorophenyl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0134] Target compound structure:

[0135] English name:

[0136] 6-(2-Chloro-6-methylphenyl)-2-(4-fluorophenyl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one

[0137] The specific steps are as follows:

[0138] Compound LWM-A24 was prepared by following the route described above. The key intermediate 6-(2-chloro-6-methylphenyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (399 mg, 1 mmol, 1 equiv), 4-fluorophenylboronic acid (280 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed sequentially in a 35 mL sealed tube and stirred at 100°C under argon for 8 hours. After completion of the reaction, the excess metal reagent was filtered through Celite to remove impurities. The organic phase was concentrated, 70 mL of DCM was added, and the mixture was extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to obtain LWM-A16 as a white solid.

[0139] After the above preparation process, 210 mg of white solid was obtained with a yield of 47%.

[0140] 1 H NMR (400MHz, DMSO-d6) δ8.25 (dd, J=8.7, 5.3Hz, 2H), 7.51–7.25 (m, 10H), 2.17 (s, 3H). 13 C NMR(101MHz,DMSO-d6)δ173.09,165.83,163.39,158.53,156.17,138.53,133.80,133.59,131.78,130.79,130.40,1 29.93(d,J=9.1Hz),129.75,128.62(d,J=3.0Hz),127.87(d,J=6.6Hz),127.48,116.80(d,J=22.3Hz),115.81,18.00.

[0141] C 24 H 15 ClFN3OS high resolution mass spectrum: 430.06768.

[0142] Example 6: LYZ-20240505

[0143] This example is used to illustrate the preparation and identification of the compound 6-(2-chloro-6-methylphenyl)-5-phenyl-2-(4-pyridyl)thiazolo[4,5-d]pyrimidin-7(6H)-one.

[0144] Target compound structure:

[0145] English name:

[0146] 6-(2-chloro-6-methylphenyl)-5-phenyl-2-(pyridin-4-yl)thiazolo[4,5-d]pyrimidin-7(6H)-one

[0147] The specific steps are as follows:

[0148] Prepared according to the route of compound LWM-A24. The key intermediate 6-(2-chloro-6-methylphenyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (399 mg, 1 mmol, 1 equiv), 4-pyridineboronic acid (246 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed in a 35 mL sealed tube and stirred at 100°C under argon for 48 hours. After completion of the reaction, the excess metal reagent was filtered through Celite to remove impurities. The organic phase was concentrated, 70 mL of DCM was added, and the product was extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to obtain LYZ-20240505 as a white solid.

[0149] After the above preparation process, 215 mg of white solid was obtained, with a yield of 50%.

[0150] 1 H NMR (400MHz, CDCl3) δ8.83(s,2H),8.01(s,2H),7.50(s,2H),7.38–7.17(m,5H),7.12(s,1H),2.16(s,3H). 13 C NMR (101MHz, CDCl3) δ172.4,166.4,159.5,156.9,151.2,139.2,138.1,134.3 ,133.6,133.0,130.5,130.5,129.6,128.5,127.9,127.9,120.8,117.2,18.6.

[0151] C 23 H 15 ClN4OS high resolution mass spectrum: 431.0718.

[0152] Example 7: LYZ-20240515

[0153] This example is used to illustrate the preparation and identification of the compound 6-(6-chloro-2-methylphenyl)-2-(2-methoxy-5-pyrimidinyl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0154] Target compound structure:

[0155] English name:

[0156] 6-(2-chloro-6-methylphenyl)-2-(2-methoxypyrimidin-5-yl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one

[0157] The specific steps are as follows:

[0158] Prepared according to the route of compound LWM-A24. The key intermediate 6-(2-chloro-6-methylphenyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (399 mg, 1 mmol, 1 equiv), 2-methoxy-5-pyrimidineboronic acid (308 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed in a 35 mL sealed tube and stirred at 100°C under argon for 48 hours. After completion of the reaction, the excess metal reagent was filtered through Celite to remove impurities. The organic phase was concentrated, 70 mL of DCM was added, and the product was extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to obtain LYZ-20240505 as a white solid.

[0159] Through the above preparation process, 208 mg of light yellow solid was obtained, with a yield of 45%.

[0160] 1 H NMR (400MHz, CDCl3) δ9.26 (s, 2H), 7.43–7.33 (m, 5H), 7.32–7.27 (m, 2H), 7.15 (d, J = 8.15Hz, 2H), 4.16 (s, 3H). 13 C NMR (101MHz, CDCl3) δ169.3,167.3,166.1,159.3,158.4,158.0,135.6,135.2,134.3,130.5,130.3,129.6,129.5,128.3,121.5,116.0,56.0.

[0161] C 23 H 16 ClN5O2S high resolution mass spectrum: 462.0786.

[0162] Example 8: WH20240505

[0163] This example is used to illustrate the preparation and identification of the compound 6-(6-chloro-2-methylphenyl)-2-(1-imidazolyl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0164] Target compound structure:

[0165] English name:

[0166] 6-(2-chloro-6-methylphenyl)-2-(1H-imidazol-1-yl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one

[0167]

[0168] The starting material is 2-chloro-6-methylaniline, which undergoes a nucleophilic substitution reaction with chloroacetyl chloride to obtain the corresponding intermediate, which is then reacted with dipotassium cyanodithiocarbamate and iodomethane in sequence to obtain the corresponding intermediate. This intermediate reacts with trimethyl orthobenzoate to obtain a key intermediate, which is oxidized with Na2WO4 and hydrogen peroxide and then undergoes a nucleophilic substitution reaction with imidazole to obtain compound WH20240505.

[0169] The specific steps are as follows:

[0170] (1) The starting material 2-chloro-6-methylaniline (14 g, 100 mmol, 1.0 equiv) was dissolved in 80 mL of saturated AcONa solution and 100 mL of AcOH solution. Chloroacetyl chloride (16.8 g, 150 mmol, 1.5 equiv) was added dropwise to the solution at room temperature, followed by stirring at room temperature for about 3 hours. After the reaction was completed, 300 mL of water was added to the system to precipitate the product. The product was filtered and the solid was washed three times with water to obtain the intermediate 2-chloro-N-(2-chloro-6-methylphenyl)acetamide.

[0171] (2) Intermediate 2-chloro-N-(2-chloro-6-methylphenyl)acetamide (10.9 g, 50 mmol, 1.0 equiv) and potassium cyanodithioimidazolide (9.7 g, 50 mmol, 1.0 equiv) were dissolved in 200 mL of acetone-water (1 / 1) mixed solution. 25 mL of 2 M aqueous NaOH solution was added. The reaction was heated to 80 °C and stirred for 1 hour. After the reaction was cooled, iodomethane (7.1 g, 50 mmol, 1.0 equiv) was added at room temperature. The system was stirred at room temperature for 30 minutes. Water was added to the system to precipitate the product, which was filtered and washed with water to obtain intermediate 4-amino-N-(2-chloro-6-methylphenyl)-2-methylthiothiazole-5- amide.

[0172] (3) Intermediate 4-amino-N-(2-chloro-6-methylphenyl)-2-methylthiothiazole-5- amide (783 mg, 2.5 mmol, 1.0 equiv) was dissolved in 6 mL of trimethyl orthobenzoate, and 0.5 mL of acetic acid was added. The reaction was heated to 120 °C and stirred for 2 days. After the reaction was completed, the reaction was added to EA, and the organic phase was extracted with saturated NaHCO3 three times. The organic phase was dried and concentrated, and then column chromatography on silica gel (dry column, PE / EA = 10 / 1 ~ 3 / 1) was performed to obtain intermediate 6-(2-chloro-6-methylphenyl)-2-methylthio-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0173] (4) Intermediate 6-(2-chloro-6-methylphenyl)-2-methylthio-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (399 mg, 1 mmol, 1 equiv) was dissolved in 5 mL of ethanol, and Na2WO4·2H2O (132 mg, 0.4 mmol, 0.2 equiv) and 2.5 mL of 30% hydrogen peroxide were sequentially added. The reaction was heated to 80 °C and stirred for 2 hours. After the reaction was completed, the reaction was added to excess ice water to precipitate the product, which was filtered to obtain a crude product. The crude product was recrystallized in ethanol to obtain key intermediate 6-(2-chloro-6-methylphenyl)-2-methylsulfonamido-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0174] (5) The intermediate 6-(2-chloro-6-methylphenyl)-2-methanesulfonyl-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (500 mg, 1.2 mmol, 1 equiv) and imidazole (160 mg, 2.4 mmol, 2 equiv) were dissolved in 30 mL of glacial acetic acid and reacted at room temperature overnight. After the reaction, cold water was added and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed three times with saturated sodium chloride solution, and then dried over anhydrous sodium sulfate. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain a crude product. The crude product was recrystallized from ethanol to obtain WH20240505 as a white solid.

[0175] Through the above preparation process, 38 mg of pale white solid was obtained, with a yield of 30%.

[0176] 1 H NMR (400MHz, CDCl3) δ8.4(s,1H),7.7(s,1H),7.5–7.4(m,2H),7.4–7.3(m,1H),7.3–7.2(m,5H),7.2–7.1(m,1H),2.2(s,3H). 13 C NMR (101MHz, CDCl3) δ164.2,162.7,160.0,156.4,138.0,134.1,133.4,133.0,132.1,130.6,130.6,129.6,128.4,127.9,127.9,112.7,18.5.

[0177] C 21 H 14 ClN5OS high resolution mass spectrum: 420.06071.

[0178] Example 9: WH20240508

[0179] This example is used to illustrate the preparation and identification of the compound 6-(6-chloro-2-methylphenyl)-2-morpholinyl-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0180] Target compound structure:

[0181] English name:

[0182] 6-(2-chloro-6-methylphenyl)-2-morpholino-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one

[0183] The specific steps are as follows:

[0184] Reference compound WH20240505 was prepared. 6-(2-Chloro-6-methylphenyl)-2- methanesulfonamido-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (500 mg, 1.2 mmol, 1 equiv) and morpholine (1174 mg, 2.4 mmol, 2 equiv) were dissolved in 30 mL glacial acetic acid and reacted at room temperature overnight. After the reaction was completed, cold water was added, and the mixture was extracted with ethyl acetate three times, and the organic phases were combined, washed with saturated sodium chloride solution three times, and then dried over anhydrous sodium sulfate. After the organic phase was concentrated, it was purified by silica gel column chromatography (PE / EA = 10 / 1 ~ 3 / 1) to obtain a crude product. The crude product was recrystallized with ethanol to obtain white solid WH20240508.

[0185] By the above preparation process, 176 mg of light green solid was obtained, with a yield of 40%.

[0186] 1 H NMR (400 MHz, CDC13) δ 7.4 (d, J = 7.5 Hz, 2H), 7.3 - 7.1 (m, 2H), 7.1 (d, J = 7.5 Hz, 1H), 3.9 (t, J = 4.8 Hz, 2H), 3.7 (t, J = 4.9 Hz, 2H), 2.1 (s, 1H). 13 C NMR (101 MHz, CDC13) δ 173.7, 166.2, 158.8, 156.1, 138.2, 134.7, 134.1, 133.2, 130.1, 130.0, 129.3, 128.4, 127.7, 127.6, 107.0, 66.1, 48.3, 18.5.

[0187] C 22 H 19 ClN4O2S High resolution mass: 439.09179.

[0188] Example 10: LWM-C4(S)

[0189] This example is used to illustrate the preparation and identification of compound 6-(2- methylphenyl)-2-(2-thienyl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0190] Structure of the target compound:

[0191] English name:

[0192] 5-Phenyl-2-(thiophen-2-yl)-6-(o-tolyl)thiazolo[4,5-d]pyrimidin-7(6H)-one

[0193] The specific steps are as follows:

[0194] The starting material is o-toluidine, and the preparation route refers to compound LWM-A24. 6-(2-methylphenyl)-2-methylthio-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (365 mg, 1 mmol, 1 equiv), thiophene-2-boronic acid (256 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), Pd(PPh3)4(58 mg, 0.05 mmol, 0.05 equiv) were sequentially placed in a 35 mL sealed tube, and stirred at 100°C for 48 hours under argon protection. After the reaction was completed, the excess metal reagent was removed by filtration with diatomite. After the organic phase was concentrated, 70 mL of DCM was added, and extracted with 10% NaHCO3three times. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 ~ 3 / 1) to obtain the crude product. The crude product was recrystallized with ethanol to obtain light green solid LWM-C4(S).

[0195] Through the above preparation process, 237 mg of light green solid was obtained, and the yield was 59%.

[0196] 1 H NMR (400 MHz, DMSO-d6) δ 8.08 (d, J = 3.6 Hz, 1H), 8.01 (d, J = 4.9 Hz, 1H), 7.45-7.20 (m, 9H), 7.20-7.11 (m, 1H), 2.07 (s, 3H). 13 C NMR (101 MHz, DMSO-d6) δ 167.51, 165.88, 159.43, 157.45, 136.77, 135.97, 135.79, 134.99, 133.32, 131.32, 131.04, 130.43, 130.11, 129.65, 129.59, 129.07, 128.10, 126.92, 115.61, 17.80.

[0197] C 22 H 15 N3OS2 high resolution mass spectrum: 402.07253.

[0198] Example 11: LWM-C28

[0199] This example is used for the preparation and identification of 6-(2-ethylphenyl)-2-(2-thienyl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0200] The structure of the target compound is: The structure of the target compound is:

[0201] English name:

[0202] 6-(2-Ethylphenyl)-5-phenyl-2-(thiophen-2-yl)thiazolo[4,5-d]pyrimidin-7(6H)-one

[0203] The specific steps are as follows:

[0204] The starting material was 2-ethylaniline, and the preparation route followed that of compound LWM-A24. The key intermediate, 6-(2-chlorophenyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (379 mg, 1 mmol, 1 equiv), thiophene-2-phenylboronic acid (256 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed in a 35 mL sealed tube and stirred at 100°C under argon for 48 hours. After completion of the reaction, the excess metal reagent was filtered through celite to remove impurities. The organic phase was concentrated, 70 mL of DCM was added, and the product was extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to obtain LWM-C28 as a white solid.

[0205] Through the above preparation process, 249 mg of light green solid was obtained, with a yield of 60%.

[0206] 1 H NMR(400MHz, CDCl3) δ7.80(d,J=2.9Hz,1H),7.59(d,J=4.3Hz,1H),7.41–7.34(m,2 H),7.34–7.23(m,3H),7.23–7.11(m,5H),2.53–2.25(m,2H),1.12(t,J=7.5Hz,3H). 13 CNMR(101MHz,CDCl3)δ168.33,166.02,159.21,158.00,140.79,136.56,135.63,134.26,131 .38,129.86,129.63,129.52,129.25,128.91,128.42,127.65,126.58,115.42,23.58,13.24.

[0207] C 23 H 17 N3OS2 high-resolution mass spectrum: 416.08792.

[0208] Example 12: LWM-C22

[0209] This example is used to illustrate the preparation and identification of the compound 6-(4-fluorophenyl)-2-(2-thienyl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0210] Target compound structure:

[0211] English name:

[0212] 6-(4-Fluorophenyl)-5-phenyl-2-(thiophen-2-yl)thiazolo[4,5-d]pyrimidin-7(6H)-one

[0213] The specific steps are as follows:

[0214] The starting material was p-fluoroaniline, and the preparation route followed that of compound LWM-A24. The key intermediate, 6-(4-fluorophenyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (369 mg, 1 mmol, 1 equiv), thiophene-2-phenylboronic acid (256 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed in a 35 mL sealed tube and stirred at 100°C under argon for 48 hours. After completion of the reaction, the excess metal reagent was filtered through celite to remove impurities. The organic phase was concentrated, 70 mL of DCM was added, and the product was extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to obtain a light green solid LWM-C22.

[0215] Through the above preparation process, 230 mg of light green solid was obtained, with a yield of 57%.

[0216] 1 H NMR (400MHz, CDCl3) δ7.79(d,J=3.4Hz,1H),7.59(d,J=4.9Hz,1H),7.36(d,J=7.3Hz,2H),7.32–7.27(m,1H),7.27–7.12(m,5H),7.02(t,J=8.4Hz,2H). 13CNMR (101 MHz, CDC13) δ 168.44, 165.68, 163.43, 160.94, 158.93, 157.99, 136.39, 134.36, 132.98 (d, J = 3.4 Hz), 131.46, 130.86 (d, J = 8.8 Hz), 129.79, 129.63, 129.33, 128.42, 127.93, 116.14 (d, J = 23.1 Hz), 115.32, 77.38, 77.06, 76.74.

[0217] C 21 H 12 FN3OS2 High resolution mass: 406.04761.

[0218] Example 13: LWM-C11

[0219] This example is used to illustrate the preparation and identification of compound 6-(4-chlorophenyl)-2-(2-thienyl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0220] Structure of the target compound:

[0221] English name:

[0222] 6-(4-Chlorophenyl)-5-phenyl-2-(thiophen-2-yl)thiazolo[4,5-d]pyrimidin-7(6H)-one

[0223] The specific steps are as follows:

[0224] The starting material is p-chloroaniline, and the preparation route refers to compound LWM-A24. The key intermediate 6-(4-fluorophenyl)-2-methylthio-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (385 mg, 1 mmol, 1 equiv), thiophene-2-phenylboronic acid (256 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), Pd(PPh3)4(58 mg, 0.05 mmol, 0.05 equiv) were sequentially placed in a 35 mL sealed tube, and stirred at 100°C under argon protection for 48 hours. After the reaction was completed, the excess metal reagent was removed by filtration with diatomite. After the organic phase was concentrated, 70 mL of DCM was added, and extracted with 10% NaHCO3three times. After the organic phase was concentrated, it was purified by silica gel column chromatography (PE / EA = 10 / 1 ~ 3 / 1) to obtain the crude product. The crude product was recrystallized with ethanol to obtain light green solid LWM-C11.

[0225] Through the above preparation process, 248 mg of light green solid was obtained, with a yield of 59%.

[0226] 1 H NMR (400MHz, DMSO-d6) δ8.07 (d, J = 3.7Hz, 1H), 8.03–7.96 (m, 1H), 7.47–7.36 (m, 6H), 7.36–7.25 (m, 4H). 13 C NMR(101MHz,DMSO-d6)δ167.41,165.69,159.31,157.93,136.71,135.96,135.27, 133.66,133.31,131.82,131.33,129.94,129.65,129.48,129.20,128.21,115.54.

[0227] C 21 H 12 ClN3OS2 high resolution mass spectrum: 422.01807.

[0228] Example 14: LWM-C12

[0229] This example is used to illustrate the preparation and identification of the compound 6-(4-methylphenyl)-2-(2-thienyl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0230] Target compound structure:

[0231] English name:

[0232] 5-Phenyl-2-(thiophen-2-yl)-6-(p-tolyl)thiazolo[4,5-d]pyrimidin-7(6H)-one

[0233] The specific steps are as follows:

[0234] The starting material was p-toluidine, and the preparation route followed that of compound LWM-A24. The key intermediate, 6-(4-methylphenyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (365 mg, 1 mmol, 1 equiv), thiophene-2-phenylboronic acid (256 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed in a 35 mL sealed tube and stirred at 100°C under argon for 48 hours. After completion of the reaction, the excess metal reagent was filtered through celite to remove impurities. The organic phase was concentrated, 70 mL of DCM was added, and the product was extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to obtain a light green solid LWM-C12.

[0235] After the above preparation process, 220 mg of light green solid was obtained, with a yield of 55%.

[0236] 1 H NMR (400MHz, DMSO-d6) δ8.06(d,J=3.6Hz,1H),7.99(d,J=4.9Hz,1H),7.40(d,J=7.4Hz,2H),7.35–7.19(m,6H),7.13(d,J=7.9Hz,2H),2.26(s,3H). 13 CNMR(101MHz,DMSO-d6)δ167.30,165.64,159.63,158.08,138.43,136.02,135.56 ,135.10,133.21,131.23,129.76,129.64,129.60,129.44,128.07,115.57,21.12.

[0237] C 22 H 15 N3OS2 high resolution mass spectrum: 402.07245.

[0238] Example 15: LWM-C10

[0239] This example is used to illustrate the preparation and identification of the compound 6-(2,6-dimethylphenyl)-2-(2-thienyl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0240] Target compound structure:

[0241] English name:

[0242] 6-(2,6-Dimethylphenyl)-5-phenyl-2-(thiophen-2-yl)thiazolo[4,5-d]pyrimidin-7(6H)-one

[0243] The specific steps are as follows:

[0244] The starting material was 2,6-dimethylaniline, and the preparation route followed that of compound LWM-A24. The key intermediate, 6-(2,6-dimethylphenyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (379 mg, 1 mmol, 1 equiv), thiophene-2-phenylboronic acid (256 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed in a 35 mL sealed tube and stirred at 100°C under argon for 48 hours. After completion of the reaction, the excess metal reagent was filtered through celite to remove impurities. The organic phase was concentrated, 70 mL of DCM was added, and the product was extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to obtain a light green solid LWM-C10.

[0245] Through the above preparation process, 237 mg of light green solid was obtained, with a yield of 57%.

[0246] 1 H NMR (400MHz, CDCl3) δ7.83(d,J=3.6Hz,1H),7.62(d,J=4.9Hz,1H),7.42(d,J=7.8Hz ,2H),7.32(t,J=7.3Hz,1H),7.25–7.15(m,4H),7.07(d,J=7.6Hz,2H),2.12(s,6H). 13 C NMR (101MHz, CDCl3) δ168.41,166.30,159.01,157.21,136.57,135.73,135.57,133. 93,131.43,130.21,129.57,129.42,128.85,128.72,128.45,127.64,115.39,18.23.

[0247] C 23 H 17 N3OS2 high-resolution mass spectrum: 416.08811.

[0248] Example 16: LWM-C29

[0249] This example is used to illustrate the preparation and identification of the compound 6-(2-methyl-3-pyridyl)-2-(2-thienyl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0250] Target compound structure:

[0251] English name:

[0252] 6-(2-Methylpyridin-3-yl)-5-phenyl-2-(thiophen-2-yl)thiazolo[4,5-d]pyrimidin-7(6H)-one

[0253] The specific steps are as follows:

[0254] The starting material was 2-methyl-3-aminopyridine, and the preparation route followed that of compound LWM-A24. The key intermediate, 6-(2-methyl-3-pyridyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (366 mg, 1 mmol, 1 equiv), thiophene-2-phenylboronic acid (256 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed in a 35 mL sealed tube and stirred at 100°C under argon for 48 hours. After completion of the reaction, the excess metal reagent was filtered through celite to remove impurities. The organic phase was concentrated, 70 mL of DCM was added, and the product was extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to obtain a light green solid LWM-C29.

[0255] After the above preparation process, 12 mg of light green solid was obtained, with a yield of 3%.

[0256] 1 H NMR (400MHz, DMSO-d6) δ8.39(dd,J=4.7,1.4Hz,1H),8.08(dd,J=3.8,0.9Hz,1H),8.01(dd,J=5.0,0.9Hz,1H) ,7.83(dd,J=8.0,1.4Hz,1H),7.43–7.36(m,2H),7.36–7.26(m,4H),7.24(dd,J=7.9,4.8Hz,1H),2.27(s,3H). 13C NMR(101MHz,DMSO-d6)δ167.20,165.47,158.64,156.91,155.30,149.48,137.94,135.42, 134.20,132.98,132.80,130.97,129.87,129.21,128.63,127.88,121.78,115.11,20.51.

[0257] C 21 H 14 N4OS2 high resolution mass spectrum: 403.06754.

[0258] Example 17: WH20231126

[0259] This example is used to illustrate the preparation and identification of the compound 5,6-diphenyl-2-(5-pyrimidinyl)thiazolo[4,5-d]pyrimidin-7(6H)-one.

[0260] Target compound structure:

[0261] English name: 5,6-diphenyl-2-(pyrimidin-5-yl)thiazolo[4,5-d]pyrimidin-7(6H)-one

[0262] The specific steps are as follows:

[0263] The starting material was aniline, and the preparation route followed compound LWM-A24. The key intermediate, 2-methylmercapto-5,6-diphenylthiazolo[4,5-d]pyrimidin-7(6H)-one (351 mg, 1 mmol, 1 equiv), 5-pyrimidineboronic acid (248 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed in a 35 mL sealed tube and stirred at 100°C under argon for 48 hours. After completion of the reaction, the excess metal reagent was filtered through celite to remove impurities. The organic phase was concentrated, added with 70 mL of DCM, and extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to yield WH20231126 as a light green solid.

[0264] Through the above preparation process, 228 mg of light green solid was obtained, with a yield of 42%.

[0265] 1H NMR (400MHz, DMSO-d6) δ9.55(s,2H),9.43(s,1H),7.44–7.21(m,10H). 13 C NMR (101MHz, DMSO-d6) δ167.1,161.9,159.1,157.5,154.9,154.4,137.3,135.7,130.5,129.9,129.3,129.3,128.4,128.3,125.0,117.8.

[0266] C 21 H 13 N5OS high-resolution mass spectrum: 384.08492.

[0267] Example 18: WH20230901

[0268] This example is used to illustrate the preparation and identification of the compound 5-phenyl-2-(5-pyrimidinyl)-6-(o-tolyl)thiazolo[4,5-d]pyrimidin-7(6H)-one.

[0269] Target compound structure:

[0270] English name: 5-phenyl-2-(pyrimidin-5-yl)-6-(o-tolyl)thiazolo[4,5-d]pyrimidin-7(6H)-one

[0271] The specific steps are as follows:

[0272] The starting material was o-toluidine, and the preparation route followed that of compound LWM-A24. The key intermediate, 6-(o-tolyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (365 mg, 1 mmol, 1 equiv), 5-pyrimidineboronic acid (248 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed in a 35 mL sealed tube and stirred at 100°C under argon for 48 hours. After completion of the reaction, the excess metal reagent was filtered through celite to remove impurities. The organic phase was concentrated, 70 mL of DCM was added, and the product was extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to obtain a light green solid WH20230901.

[0273] Through the above preparation process, 155 mg of light green solid was obtained, with a yield of 39%.

[0274] 1 H NMR (400MHz, DMSO-d6) δ9.55(s,2H),9.44(s,1H),7.42–7.15(m,9H),2.08(s,3H). 13 C NMR (101MHz, DMSO-d6) δ168.4,166.0,161.0,159.6,157.6,155.7,131.1,130.4,130.2,129.7,129.1,128.2,127.0,117.5,17.8.

[0275] C 22 H 15 N5OS high-resolution mass spectrum: 398.09977.

[0276] Example 19: WH20231215

[0277] This example is used to illustrate the preparation and identification of the compound 6-(4-chlorophenyl)-5-phenyl-2-(5-pyrimidinyl)thiazolo[4,5-d]pyrimidin-7(6H)-one.

[0278] Target compound structure:

[0279] English name:

[0280] 6-(4-chlorophenyl)-5-phenyl-2-(pyrimidin-5-yl)thiazolo[4,5-d]pyrimidin-7(6H)-one

[0281] The specific steps are as follows:

[0282] The starting material was p-chloroaniline, and the preparation route followed that of compound LWM-A24. The key intermediate, 6-(4-chlorophenyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (385 mg, 1 mmol, 1 equiv), 5-pyrimidineboronic acid (248 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed in a 35 mL sealed tube and stirred at 100°C under argon for 48 hours. After completion of the reaction, the excess metal reagent was filtered through celite to remove impurities. The organic phase was concentrated, 70 mL of DCM was added, and the product was extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to obtain a light green solid WH20231215.

[0283] Through the above preparation process, 245 mg of light green solid was obtained, with a yield of 38%.

[0284] 1 H NMR (400MHz, CDCl3) δ9.47 (s, 2H), 9.40 (s, 1H), 7.43–7.32 (m, 5H), 7.32–7.26 (m, 2H), 7.15 (d, J = 8.2Hz, 2H). 13 C NMR (101MHz, CDCl3) δ168.5,165.9,160.8,159.4,157.9,155.2,135.3,135.1,134.0,130.3,129.5,129.3,128.2,127.0,116.9.

[0285] C 21 H 12 ClN5OS high resolution mass spectrum: 418.04515.

[0286] Example 20: WH20231228

[0287] This example is used to illustrate the preparation and identification of the compound 6-(4-fluorophenyl)-5-phenyl-2-(5-pyrimidinyl)thiazolo[4,5-d]pyrimidin-7(6H)-one.

[0288] Target compound structure:

[0289] English name:

[0290] 6-(4-fluorophenyl)-5-phenyl-2-(pyrimidin-5-yl)thiazolo[4,5-d]pyrimidin-7(6H)-one

[0291] The specific steps are as follows:

[0292] The starting material was p-fluoroaniline, and the preparation route followed that of compound LWM-A24. The key intermediate, 6-(4-fluorophenyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (369 mg, 1 mmol, 1 equiv), 5-pyrimidineboronic acid (248 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed in a 35 mL sealed tube and stirred at 100°C under argon for 48 hours. After completion of the reaction, the excess metal reagent was filtered through celite to remove impurities. The organic phase was concentrated, 70 mL of DCM was added, and the product was extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to obtain a light green solid WH20231228.

[0293] Through the above preparation process, 140 mg of light green solid was obtained, with a yield of 35%.

[0294] Example 21: WH20240413

[0295] This example is used to illustrate the preparation and identification of the compound 2-(2-methoxy-5-pyrimidinyl)-6-(o-tolyl)thiazolo[4,5-d]pyrimidin-7(6H)-one.

[0296] Target compound structure:

[0297] English name:

[0298] 2-(2-methoxypyrimidin-5-yl)-5-phenyl-6-(o-tolyl)thiazolo[4,5-d]pyrimidin-7(6H)-one

[0299] The specific steps are as follows:

[0300] The starting material was o-toluidine, and the preparation route followed that of compound LWM-A24. The key intermediate, 6-(o-tolyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (365 mg, 1 mmol, 1 equiv), 2-methoxy-5-pyrimidineboronic acid (308 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed in a 35 mL sealed tube and stirred at 100°C under argon for 48 hours. After completion of the reaction, the excess metal reagent was filtered through celite to remove impurities. The organic phase was concentrated, 70 mL of DCM was added, and the product was extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to obtain a light green solid WH20240413.

[0301] After the above preparation process, 150 mg of light green solid was obtained with a yield of 35%.

[0302] 1 H NMR (400MHz, CDCl3) δ9.23 (s, 2H), 7.38 (d, J = 7.6Hz, 2H), 7.31–7.06 (m, 7H), 4.11 (s, 3H), 2.13 (s, 3H). 13 C NMR (101MHz, CDCl3) δ168.9,167.1,166.2,159.5,158.2,157.5,136.2,135.6,13 4.1,131.2,130.1,129.5,129.4,129.0,127.8,126.9,121.4,115.9,55.8,17.9.

[0303] C 23 H 17 N5O2S high resolution mass spectrum: 428.11035.

[0304] Example 22: LYZ-20240630

[0305] This example is used to illustrate the preparation and identification of the compound 6-(4-chlorophenyl)-2-(2-methoxy-5-pyrimidinyl)thiazolo[4,5-d]pyrimidin-7(6H)-one.

[0306] Target compound structure:

[0307] English name:

[0308] 6-(4-chlorophenyl)-2-(2-methoxypyrimidin-5-yl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one

[0309] The specific steps are as follows:

[0310] The starting material was p-chloroaniline, and the preparation route followed that of compound LWM-A24. The key intermediate, 6-(4-fluorophenyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (385 mg, 1 mmol, 1 equiv), 2-methoxy-5-pyrimidineboronic acid (308 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed in a 35 mL sealed tube and stirred at 100°C under argon for 48 hours. After completion of the reaction, the excess metal reagent was filtered through celite to remove impurities. The organic phase was concentrated, 70 mL of DCM was added, and the product was extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to obtain a light yellow solid LYZ-20240630.

[0311] Through the above preparation process, 179 mg of light yellow solid was obtained, with a yield of 40%.

[0312] 1 H NMR (400MHz, CDCl3) δ9.26 (s, 2H), 7.43–7.33 (m, 5H), 7.32–7.27 (m, 2H), 7.15 (d, J = 8.15Hz, 2H), 4.16 (s, 3H). 13 C NMR (101MHz, CDCl3) δ169.3,167.3,166.1,159.3,158.4,158.0,135.6,135.2,134.3,130.5,130.3,129.6,129.5,128.3,121.5,116.0,56.0.

[0313] C 22 H 14 ClN5O2S high resolution mass spectrum: 448.0618.

[0314] Example 23: LYZ-20240716

[0315] This example is used to illustrate the preparation and identification of the compound 6-(4-fluorophenyl)-2-(2-methoxy-5-pyrimidinyl)thiazolo[4,5-d]pyrimidin-7(6H)-one.

[0316] Target compound structure:

[0317] English name:

[0318] 6-(4-fluorophenyl)-2-(2-methoxypyrimidin-5-yl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one

[0319] The specific steps are as follows:

[0320] The starting material was p-fluoroaniline, and the preparation route followed that of compound LWM-A24. The key intermediate, 6-(4-fluorophenyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (369 mg, 1 mmol, 1 equiv), 2-methoxy-5-pyrimidineboronic acid (308 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed in a 35 mL sealed tube and stirred at 100°C under argon for 48 hours. After completion of the reaction, the excess metal reagent was filtered through celite to remove impurities. The organic phase was concentrated, 70 mL of DCM was added, and the product was extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to obtain a light yellow solid LYZ-20240716.

[0321] Through the above preparation process, 179 mg of light yellow solid was obtained, with a yield of 40%.

[0322] 1 H NMR (400MHz, CDCl3) δ9.26(s,2H),7.46–7.37(m,2H),7.37–7.31(m,1H),7.31–7.23(m,2H),7.22–7.15(m,2H),7.11–7.01(m,2H),4.16(s,3H). 13C NMR (101MHz, CDCl3) δ169.3, 167.3, 166.1, 162.5 (d, J = 250.2Hz), 159.5, 158.4, 158.2, 134.4, 133. 0(d,J=3.5Hz),130.9(d,J=8.9Hz),130.2,129.5,128.3,121.5,116.5(d,J=23.1Hz),116.0,56.0.

[0323] C 22 H 14 FN5O2S high resolution mass spectrum: 432.08527.

[0324] Example 24: LYZ-20240717

[0325] This example is used to illustrate the preparation and identification of the compound 2-(2-methoxy-5-pyrimidinyl)-5-phenyl-6-(p-tolyl)thiazolo[4,5-d]pyrimidin-7(6H)-one.

[0326] Target compound structure:

[0327] English name:

[0328] 2-(2-methoxypyrimidin-5-yl)-5-phenyl-6-(p-tolyl)thiazolo[4,5-d]pyrimidin-7(6H)-one

[0329] The specific steps are as follows:

[0330] The starting material was p-toluidine, and the preparation route followed that of compound LWM-A24. The key intermediate, 6-(p-tolyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (365 mg, 1 mmol, 1 equiv), 2-methoxy-5-pyrimidineboronic acid (308 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed in a 35 mL sealed tube and stirred at 100°C under argon for 48 hours. After completion of the reaction, the excess metal reagent was filtered through Celite to remove impurities. The organic phase was concentrated, 70 mL of DCM was added, and the product was extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to obtain a light yellow solid LYZ-20240717.

[0331] Through the above preparation process, 188 mg of light yellow solid was obtained, with a yield of 44%.

[0332] 1 H NMR (400MHz, CDCl3) δ9.27 (s, 2H), 7.42 (d, J = 7.55Hz, 2H), 7.35–7.24 (m, 3H ),7.17(d,J=7.83Hz,2H),7.08(d,J=7.84Hz,2H),4.16(s,3H),2.36(s,3H). 13 CNMR(101MHz,CDCl3)δ169.0,167.2,166.1,159.7,158.3,139.2,134.7,1 34.4,130.0,130.0,129.5,128.8,128.0,121.6,121.6,116.1,55.9,21.3.

[0333] C 23 H 17 N5O2S high resolution mass spectrum: 428.11036.

[0334] Example 25: WH20240604

[0335] This example is used to illustrate the preparation and identification of the compound 2-(2-methoxy-5-pyrimidinyl)-6-(4-nitro)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0336] Target compound structure:

[0337] English name:

[0338] 2-(2-methoxypyrimidin-5-yl)-6-(4-nitrophenyl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one

[0339] The specific steps are as follows:

[0340] The starting material was p-nitroaniline, and the preparation route followed that of compound LWM-A24. The key intermediate, 6-(4-nitrophenyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (396 mg, 1 mmol, 1 equiv), 2-methoxy-5-pyrimidineboronic acid (308 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed in a 35 mL sealed tube and stirred at 100°C under argon for 48 hours. After completion of the reaction, the excess metal reagent was filtered through celite to remove impurities. The organic phase was concentrated, 70 mL of DCM was added, and the product was extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to obtain a yellow solid WH20240604.

[0341] Through the above preparation process, 136 mg of yellow solid was obtained, with a yield of 33%.

[0342] 1 H NMR (400MHz, DMSO-d6) δ9.4(s,2H),8.2(d,J=8.4Hz,2H),7.7(d,J=8.4Hz,2H),7.5–7.4(m,2H),7.4–7.2(m,3H),4.1(s,3H). 13 C NMR (101MHz, DMSO-d6) δ167.6,166.0,161.8,157.5,154.8,154.3,142.5,141.5,135.8,130.5,129.9,129.3,126.8,125.4,120.3,117.8,54.3.

[0343] C 22 H 14 N5O2S high resolution mass spectrum: 413.08688.

[0344] Example 26: WH20240720

[0345] This example is used to illustrate the preparation and identification of the compound 6-(3,5-difluorophenyl)-(2-methoxy-5-pyrimidinyl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0346] Target compound structure:

[0347] English name:

[0348] 6-(3,5-difluorophenyl)-2-(2-methoxypyrimidin-5-yl)-5-phenylthiazolo[4,5-d]pyrimidi n-7(6H)-one

[0349] The specific steps are as follows:

[0350] (1) The starting material 3,5-difluoroaniline (13 g, 100 mmol, 1.0 equiv) was dissolved in 80 mL of saturated AcONa solution and 100 mL of AcOH solution. Chloroacetyl chloride (16.8 g, 150 mmol, 1.5 equiv) was added dropwise to the solution at room temperature, followed by stirring at room temperature for approximately 3 hours. After the reaction was completed, 300 mL of water was added to the system to precipitate the product. The product was filtered and the solid was washed three times with water to obtain the intermediate 2-chloro-N-(3,5-difluorophenyl)acetamide.

[0351] (2) The intermediate 2-chloro-N-(3,5-difluorophenyl)acetamide (10.3 g, 50 mmol, 1.0 equiv) and dipotassium cyanodithiocarbamate (9.7 g, 50 mmol, 1.0 equiv) were dissolved in 200 mL of a 1 / 1 acetone-water mixture. 25 mL of a 2 M NaOH aqueous solution was added. The reaction solution was heated to 80°C and stirred for 1 hour. After the reaction solution was cooled, iodomethane (7.1 g, 50 mmol, 1.0 equiv) was added at room temperature. The mixture was stirred at room temperature for 30 minutes. Water was added to the system to precipitate the product, which was filtered and washed with water to obtain the intermediate 4-amino-N-(3,5-difluorophenyl)-2-methylmercaptothiazole-5-carboxamide.

[0352] (3) The intermediate 4-amino-N-(3,5-difluorophenyl)-2-methylmercaptothiazole-5-carboxamide (753 mg, 2.5 mmol, 1.0 equiv) was dissolved in 6 mL of trimethyl orthobenzoate, and 0.5 mL of acetic acid was added. The reaction was heated to 120°C and stirred for 2 days. After the reaction was completed, EA was added to the reaction solution, and the organic phase was extracted three times with saturated NaHCO3. The organic phase was dried and concentrated, and then separated by silica gel column chromatography (dry column loading, PE / EA = 10 / 1 to 3 / 1) to obtain the key intermediate 6-(3,5-difluorophenyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0353] (4) The key intermediate 6-(3,5-difluorophenyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (387 mg, 1 mmol, 1 equiv), 2-methoxy-5-pyrimidineboronic acid (308 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed in a 35 mL sealed tube and stirred at 100°C for 48 hours under argon protection. After the reaction was completed, the excess metal reagent was filtered and removed with celite. The organic phase was concentrated, 70 mL of DCM was added, and the mixture was extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA=10 / 1 to 3 / 1) to obtain a crude product. The crude product was recrystallized from ethanol to obtain a light yellow solid WH20240720.

[0354] Through the above preparation process, 180 mg of light yellow solid was obtained, with a yield of 40%.

[0355] 1 H NMR (400MHz, DMSO-d6) δ9.4(s,2H),7.5–7.4(m,2H),7.4–7.2(m,6H),4.1(s,3H). 13 C NMR (101MHz, DMSO-d6) δ168.9, 167.0, 162.2 (dd, J=246.7, 14.5Hz), 159.1, 158.9, 157.7, 140.1 (t, J= 13.5Hz), 134.9, 130.2, 129.3, 128.3, 121.5, 116.2, 114.4 (d, J = 27.5Hz), 105.2 (t, J = 25.6Hz), 56.0.

[0356] C 22 H 13 F2N5O2S high resolution mass spectrum: 450.07583.

[0357] Example 27: WH20240709

[0358] This example is used to illustrate the preparation and identification of the compound 6-(2,6-dimethylphenyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0359] Target compound structure:

[0360] English name:

[0361] 6-(2,6-dimethylphenyl)-2-(2-methoxypyrimidin-5-yl)-5-phenylthiazolo[4,5-d]pyrimid in-7(6H)-one

[0362] The specific steps are as follows:

[0363] The starting material was 2,6-dimethylaniline, and the preparation route followed that of compound LWM-A24. The key intermediate, 6-(2,6-dimethylphenyl)-2-methylmercapto-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one (379 mg, 1 mmol, 1 equiv), 2-methoxy-5-pyrimidineboronic acid (308 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed in a 35 mL sealed tube and stirred at 100°C under argon for 48 hours. After completion of the reaction, the excess metal reagent was filtered through celite to remove impurities. The organic phase was concentrated, 70 mL of DCM was added, and the product was extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to obtain a pale yellow solid WH20240709.

[0364] Through the above preparation process, 140 mg of light yellow solid was obtained, with a yield of 33%.

[0365] 1 H NMR (400MHz, CDCl3) δ9.3 (s, 2H), 7.5–7.4 (m, 2H), 7.4–7.3 (m, 1H), 7.3–7.2 (m, 3H), 7.1 (d, J = 7.6Hz, 2H), 4.1 (s, 3H), 2.1 (s, 6H). 13 C NMR (101MHz, CDCl3) δ169.0,167.1,166.5,159.4,158.2,157.1,135.5,135.5,133.8,130.4,129.5,128.8,127.7,121.5,115.9,55.8,18.2.

[0366] C 24 H 19 N5O2S high resolution mass spectrum: 442.12590.

[0367] Example 28: WH20240605

[0368] This example is used to illustrate the preparation and identification of the compound 2-(2-methoxy-5-pyrimidinyl)-5-phenyl-6-(4-pyridinyl)thiazolo[4,5-d]pyrimidin-7(6H)-one.

[0369] Target compound structure:

[0370] English name:

[0371] 2-(2-methoxypyrimidin-5-yl)-5-phenyl-6-(pyridin-4-yl)thiazolo[4,5-d]pyrimidin-7(6H)-one

[0372] The specific steps are as follows:

[0373] The starting material was 4-aminopyridine, and the preparation route followed that of compound LWM-A24. The key intermediate, 2-methylmercapto-5-phenyl-6-(4-pyridyl)thiazolo[4,5-d]pyrimidin-7(6H)-one (352 mg, 1 mmol, 1 equiv), 2-methoxy-5-pyrimidineboronic acid (308 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed in a 35 mL sealed tube and stirred at 100°C under argon for 48 hours. After completion of the reaction, the excess metal reagent was filtered through Celite to remove impurities. The organic phase was concentrated, 70 mL of DCM was added, and the product was extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to obtain a pale yellow solid WH20240605.

[0374] After the above preparation process, 124 mg of light yellow solid was obtained, with a yield of 30%.

[0375] 1 H NMR (400MHz, CDCl3) δ9.2 (s, 2H), 8.6 (d, J = 5.2Hz, 2H), 7.6–7.3 (m, 3H), 7.3–7.2 (m, 2H), 7.2 (d, J = 5.1Hz, 2H), 4.1 (s, 3H). 13 C NMR (101MHz, CDCl3) δ169.5,167.2,165.9,158.5,158.3,157.2,151.0,144.9,133.6,130.5,129.3,128.3,124.0,121.2,115.8,55.9.

[0376] C 21 H 14 N6O2S high resolution mass spectrum: 415.08992.

[0377] Example 29: WH20240621

[0378] This example is used to illustrate the preparation and identification of the compound 2-(2-methoxy-5-pyrimidinyl)-5-phenyl-6-(2-pyrimidinyl)thiazolo[4,5-d]pyrimidin-7(6H)-one.

[0379] Target compound structure:

[0380] English name:

[0381] 2-(2-methoxypyrimidin-5-yl)-5-phenyl-6-(pyrimidin-2-yl)thiazolo[4,5-d]pyrimidin-7(6H)-one

[0382] The specific steps are as follows:

[0383] The starting material was 2-aminopyrimidine, and the preparation route followed that of compound LWM-A24. The key intermediate, 2-methylmercapto-5-phenyl-6-(2-pyrimidinyl)thiazolo[4,5-d]pyrimidin-7(6H)-one (352 mg, 1 mmol, 1 equiv), 2-methoxy-5-pyrimidineboronic acid (308 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed in a 35 mL sealed tube and stirred at 100°C under argon for 48 hours. After completion of the reaction, the excess metal reagent was filtered through celite to remove impurities. The organic phase was concentrated, 70 mL of DCM was added, and the product was extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to obtain a pale yellow solid WH20240621.

[0384] Through the above preparation process, 137 mg of light yellow solid was obtained, with a yield of 33%.

[0385] 1 H NMR (400MHz, CDCl3) δ9.2 (s, 2H), 8.6 (d, J = 5.2Hz, 2H), 7.6–7.3 (m, 3H), 7.3–7.2 (m, 2H), 7.2 (d, J = 5.1Hz, 2H), 4.1 (s, 3H). 13C NMR (101MHz, CDCl3) δ169.5,167.2,165.9,158.5,158.3,157.2,151.0,144.9,133.6,130.5,129.3,128.3,124.0,121.2,115.8,55.9.

[0386] C 21 H 14 N6O2S high resolution mass spectrum: 415.08992.

[0387] Example 30: LYZ-20241015

[0388] This example is used to illustrate the preparation and identification of the compound 6-(5-chloro-2-pyrimidinyl)-2-(2-methoxy-5-pyrimidinyl)-5-phenylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0389] Target compound structure:

[0390] English name:

[0391] 6-(5-chloropyrimidin-2-yl)-2-(2-methoxypyrimidin-5-yl)-5-phenylthiazolo[4,5-d]pyri midin-7(6H)-one

[0392] The specific steps are as follows:

[0393] The starting material was p-2-amino-5-chloropyrimidine, prepared using the same route as compound LWM-A24. The key intermediate, 2-methylmercapto-5-phenyl-6-(5-chloro-2-pyrimidinyl)thiazolo[4,5-d]pyrimidin-7(6H)-one (388 mg, 1 mmol, 1 equiv), 2-methoxy-5-pyrimidineboronic acid (308 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were placed in a 35 mL sealed tube and stirred at 100°C under argon for 48 hours. After completion of the reaction, the excess metal reagent was filtered through celite to remove impurities. The organic phase was concentrated, 70 mL of DCM was added, and the product was extracted three times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the crude product. The crude product was recrystallized from ethanol to obtain a light yellow solid LYZ-20241015.

[0394] 1H NMR (400MHz, CDCl3) δ9.25 (s, 2H), 8.72 (s, 2H), 7.46 (d, J = 7.55Hz, 2H), 7.38–7.33 (m, 1H), 7.29 (d, J = 7.34Hz, 2H), 4.13 (s, 3H). 13 C NMR (101MHz, CDCl3) δ169.5,167.2,166.2,158.3,158.1,157.8,157.7,155.7,133.8,131.5,130.6,128.9,128.4,121.4,115.7,55.9.

[0395] C 20 H 12 ClN7O2S high resolution mass spectrum: 450.0523.

[0396] Example 31: LYZ-20241002

[0397] This example is used to illustrate the preparation and identification of the compound 6-(4-chlorophenyl)-5-ethyl-2-(2-methoxy-5-pyrimidinyl)thiazolo[4,5-d]pyrimidin-7(6H)-one.

[0398] Target compound structure:

[0399] English name:

[0400] 6-(4-chlorophenyl)-5-ethyl-2-(2-methoxypyrimidin-5-yl)thiazolo[4,5-d]pyrimidin-7(6H)-one

[0401] The starting material is p-chloroaniline, which undergoes a nucleophilic substitution reaction with chloroacetyl chloride to obtain the corresponding intermediate, which is then reacted with dipotassium cyanodithiocarbamate and iodomethane in sequence to obtain the corresponding intermediate. This intermediate reacts with trimethyl orthopropionate to obtain a key intermediate, which undergoes a Liebeskind–Srogl coupling reaction with 2-methoxy-5-pyrimidineboronic acid in the presence of cuprous thiophene-2-carboxylate and catalyzed by Pd(PPh3)4 to obtain compound LYZ-20241002.

[0402] The specific steps are as follows:

[0403] (1) The starting material, p-chloroaniline (12.7 g, 100 mmol, 1.0 equiv), was dissolved in 80 mL of saturated AcONa solution and 100 mL of AcOH solution. Chloroacetyl chloride (16.8 g, 150 mmol, 1.5 equiv) was added dropwise to the solution at room temperature, followed by stirring at room temperature for approximately 3 hours. After the reaction was completed, 300 mL of water was added to the system to precipitate the product. The product was filtered, and the resulting solid was washed three times with water to obtain the intermediate 2-chloro-N-(4-chlorophenyl)acetamide.

[0404] (2) The intermediate 2-chloro-N-(4-chlorophenyl)acetamide (10.2 g, 50 mmol, 1.0 equiv) and dipotassium cyanodithiocarbamate (9.7 g, 50 mmol, 1.0 equiv) were dissolved in 200 mL of a 1 / 1 acetone-water mixture. 25 mL of a 2 M NaOH aqueous solution was added. The reaction solution was heated to 80°C and stirred for 1 hour. After the reaction solution was cooled, iodomethane (7.1 g, 50 mmol, 1.0 equiv) was added at room temperature. The mixture was stirred at room temperature for 30 minutes. Water was added to the system to precipitate the product, which was filtered and washed with water to obtain the intermediate 4-amino-N-(p-chlorophenyl)-2-methylmercaptothiazole-5-carboxamide.

[0405] (3) The intermediate 4-amino-N-(p-chlorophenyl)-2-methylmercaptothiazole-5-carboxamide (749 mg, 2.5 mmol, 1.0 equiv) was dissolved in 6 mL of trimethyl orthopropionate, and 0.5 mL of acetic acid was added. The reaction was heated to 120°C and stirred for 2 days. After the reaction was completed, EA was added to the reaction solution, and the organic phase was extracted three times with saturated NaHCO3. The organic phase was dried and concentrated, and then separated by silica gel column chromatography (dry column loading, PE / EA = 10 / 1 to 3 / 1) to obtain the key intermediate 6-(4-chlorophenyl)-2-methylmercapto-5-ethylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0406] (4) The intermediate 6-(4-chlorophenyl)-2-methylmercapto-5-ethylthiazolo[4,5-d]pyrimidin-7(6H)-one (338 mg, 1 mmol, 1 equiv) was dissolved in THF, and 2-methoxy-5-pyrimidineboronic acid (154 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were added in sequence. The reaction system was replaced with argon 8 times, heated to 100°C, and stirred for 48 hours. After the reaction was completed, the excess metal reagent was filtered and removed with celite. The organic phase was concentrated, 70 mL of DCM was added, and the product was extracted 3 times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain a crude product. The crude product was recrystallized from ethanol to obtain LYZ-20241002 as a white solid.

[0407] After the above preparation process, 200 mg of white solid was obtained with a yield of 50%.

[0408] 1 H NMR (400MHz, CDCl3) δ9.20 (s, 2H), 7.55 (d, J = 8.44Hz, 2H), 7.22 (d, J = 8.35Hz, 2H), 4.11 (s, 3H), 2.49 (q, J = 7.31Hz, 2H), 1.28 (t, J = 7.34Hz, 3H). 13 C NMR (101MHz, CDCl3) δ168.9,167.1,166.2,162.5,158.3,158.2,136.0,135.0,130.6,129.6,121.4,115.2,55.9,29.6,11.1.

[0409] C 18 H 14 ClN5O2S high resolution mass spectrum: 400.05569.

[0410] Example 32: LYZ-20241012

[0411] This example is used to illustrate the preparation and identification of the compound 6-(4-chlorophenyl)-5-cyclopropyl-2-(2-methoxy-5-pyrimidinyl)thiazolo[4,5-d]pyrimidin-7(6H)-one.

[0412] Target compound structure:

[0413] English name:

[0414] 6-(4-chlorophenyl)-5-cyclopropyl-2-(2-methoxypyrimidin-5-yl)thiazolo[4,5-d]pyrimidin-7(6H)-one

[0415] The starting material is p-chloroaniline, which undergoes a nucleophilic substitution reaction with chloroacetyl chloride to obtain the corresponding intermediate, which is then reacted with dipotassium cyanodithiocarbamate and iodomethane in sequence to obtain the corresponding intermediate. The intermediate is reacted with cyclopropylcarbonyl chloride to obtain the corresponding intermediate. The intermediate is cyclized under the action of DIPEA, DMAP and N,O-Bis(trimethylsilyl)acetamide to obtain a key intermediate. The key intermediate undergoes a Liebeskind–Srogl coupling reaction with 2-methoxy-5-pyrimidineboronic acid in the presence of cuprous thiophene-2-carboxylate and catalyzed by Pd(PPh3)4 to obtain compound LYZ-20241002.

[0416] The specific steps are as follows:

[0417] (1) The starting material, p-chloroaniline (12.7 g, 100 mmol, 1.0 equiv), was dissolved in 80 mL of saturated AcONa solution and 100 mL of AcOH solution. Chloroacetyl chloride (16.8 g, 150 mmol, 1.5 equiv) was added dropwise to the solution at room temperature, followed by stirring at room temperature for approximately 3 hours. After the reaction was completed, 300 mL of water was added to the system to precipitate the product. The product was filtered, and the resulting solid was washed three times with water to obtain the intermediate 2-chloro-N-(4-chlorophenyl)acetamide.

[0418] (2) The intermediate 2-chloro-N-(4-chlorophenyl)acetamide (10.2 g, 50 mmol, 1.0 equiv) and dipotassium cyanodithiocarbamate (9.7 g, 50 mmol, 1.0 equiv) were dissolved in 200 mL of a 1 / 1 acetone-water mixture. 25 mL of a 2 M NaOH aqueous solution was added. The reaction solution was heated to 80°C and stirred for 1 hour. After the reaction solution was cooled, iodomethane (7.1 g, 50 mmol, 1.0 equiv) was added at room temperature. The mixture was stirred at room temperature for 30 minutes. Water was added to the system to precipitate the product, which was filtered and washed with water to obtain the intermediate 4-amino-N-(p-chlorophenyl)-2-methylmercaptothiazole-5-carboxamide.

[0419] (3) The intermediate 4-amino-N-(p-chlorophenyl)-2-methylmercaptothiazole-5-carboxamide (749 mg, 2.5 mmol, 1.0 equiv) was dissolved in 20 mL of toluene. Cyclopropylcarbonyl chloride (460 mg, 4.4 mmol, 1.1 equiv) and DIPEA (1550 mg, 12 mmol, 3 equiv) were added in sequence under ice-cooling. The reaction solution was heated to 50°C and stirred for 4 hours. After the reaction was completed, 60 mL of EA was added to the reaction solution, and the solution was extracted three times with 10% NaHCO3. The organic phase was dried over Na2SO4 and concentrated, and then purified by silica gel column chromatography (dry column loading, PE / EA = 10 / 1 to 3 / 1) to obtain the intermediate N-(4-chlorophenyl)-4-(cyclopropylcarbonyl)-2-(2-methoxy-5-pyrimidinyl)thiazole-5-carboxamide.

[0420] (4) The intermediate N-(4-chlorophenyl)-4-(cyclopropylcarbonyl)-2-(2-methoxy-5-pyrimidinyl)thiazole-5-carboxamide (736 mg, 2 mmol, 1 equiv) was dissolved in 10 mL of ACN. DIPEA (258 mg, 2 mmol, 1 equiv), DMAP (244 mg, 2 mmol, 1 equiv) and N,O-bis(trimethylsilyl)acetamide (4 g, 20 mmol, 10 equiv) were added in sequence at room temperature. The reaction solution was heated to 80°C and stirred for 30 minutes. After the reaction was completed, the reaction solution was concentrated and 70 mL of EA was added. The product was extracted three times with brine. The organic phase was dried over Na2SO4 and concentrated, and then purified by silica gel column chromatography (dry column loading, PE / EA = 10 / 1 to 3 / 1) to obtain the key intermediate 6-(4-chlorophenyl)-2-methylmercapto-5-cyclopropylthiazolo[4,5-d]pyrimidin-7(6H)-one.

[0421] (5) The key intermediate 6-(4-chlorophenyl)-2-methylmercapto-5-cyclopropylthiazolo[4,5-d]pyrimidin-7(6H)-one (350 mg, 1 mmol, 1 equiv) was dissolved in THF, and 2-methoxy-5-pyrimidineboronic acid (154 mg, 2 mmol, 2 equiv), CuTC (380 mg, 2 mmol, 2 equiv), and Pd(PPh3)4 (58 mg, 0.05 mmol, 0.05 equiv) were added in sequence. The reaction system was replaced with argon 8 times, heated to 100°C, and stirred for 48 hours. After the reaction was completed, the excess metal reagent was filtered and removed with celite. The organic phase was concentrated, 70 mL of DCM was added, and the product was extracted 3 times with 10% NaHCO3. The organic phase was concentrated and purified by silica gel column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain a crude product. The crude product was recrystallized from ethanol to obtain LYZ-20241002 as a white solid.

[0422] After the above preparation process, 177 mg of white solid was obtained, with a yield of 43%.

[0423] 1 H NMR (400MHz, CDCl3) δ9.20 (s, 2H), 7.56 (d, J = 8.08Hz, 2H), 7.31 (d, J = 8.09Hz, 2H), 4.11 (s, 3H), 1.55–1.24 (m, 2H), 1.11–0.79 (m, 2H). 13 C NMR (101MHz, CDCl3) δ168.9,167.1,166.5,163.6,158.3,158.2,135.9,135.2,130.5,129.9,121.5,114.3,55.9,15.1,12.2.

[0424] C 19 H 14 ClN5O2S high resolution mass spectrum: 412.0619.

[0425] Test example

[0426] Test Example 1: In vitro electrophysiological activity test method

[0427] Recombinant GABA A Receptor expression in Xenopus oocytes:

[0428] Stage V-VI Xenopus oocytes were selected and microinjected with in vitro transcribed capped cRNA using the T3mMESSAGE mMACHINE Kit (Ambion). The injection volume was 23 nanoliters per oocyte, with 6 ng of total RNA and a subunit ratio of α:β:δ = 5:1:5. Following injection, oocytes were incubated at 16°C in ND96 medium (96 mM NaCl, 2 mM KCl, 1.8 mM CaCl2, 1 mM MgCl2, 5 mM HEPES, pH 7.4 adjusted with NaOH). Two-electrode voltage-clamp recordings were performed 24-72 hours after injection using a GeneClamp 500B amplifier (Axon Instruments).

[0429] Data Records:

[0430] The whole-cell current of Xenopus oocytes was recorded by two-electrode voltage clamp (TEVC) technique, and the current was recorded by GeneClamp 500B amplifier. The recording electrodes were prepared using a 1440A converter and Clampex software. Recording electrodes (resistance 0.5-1.5 MΩ) were prepared using a PC-10 puller and filled with 3 M KCl solution. After insertion into the oocyte, a -70 mV clamping voltage was applied. The external fluid was ND96 solution, perfused at a rate of 2-3 mL / min.

[0431] Drug activity testing process:

[0432] Dilute the DMSO stock solution and GABA to the working concentration using ND96. First, apply the GABA solution to record the basal current. Wash with ND96 for 2 minutes to restore the receptor state. Then, apply the drug-GABA mixture to record the action current. All operations were performed at 22 ± 1°C.

[0433] Data processing:

[0434] In the statistical analysis of the data, Clampfit software was also used to directly read the GABA current size before drug administration Icontrol and the GABA current size after drug administration Icompound. The compound activity was expressed by the following expression: Potentiation = (I compound –I control ) / I control × 100%. Graphpad Prism 8 software was used to analyze the data and plot the data. The data were expressed as mean ± standard error (mean ± SEM). Paired T-test was used to compare significant differences. P < 0.05 was considered statistically significant. The dose-effect curve of the compound was fitted using the four-parameter Hill equation in Graphpad Prism 8 software: Y = Bottom + (Top-Bottom) / (1 + 10 lgEC50-X × HillSlope), where Y represents the ratio of the current after drug administration to the current before drug administration, Icompound / Icontrol; Top represents the maximum effect; Bottom represents the minimum effect; X is the logarithm of drug concentration; EC 50 It is the drug concentration value corresponding to the half-maximal effect; HillSlope represents the slope of the curve.

[0435] The dose-effect curves of representative compounds (WH20240720, WH20240413 and LY-0240630), positive drug allopregnanolone and positive compound DS-2 in Xenopus oocytes expressing α4β3δ are shown in Figure 2. Figure 1 shown.

[0436] At the same time, the current allosteric enhancement activity data of representative compounds (at 10 micromolar concentration) and EC 50 The values ​​(half maximal effect concentrations) are listed in Table 1 below.

[0437] Table 1 In vitro electrophysiological activity results

[0438]

[0439]

[0440] a Data from 3-5 Xenopus oocytes expressing human α4β3δ GABA A Receptor two-electrode voltage clamp current. EC 50 The half-maximal effect concentration is the concentration of the test compound required to induce 50% of the maximum current and is used to measure the potency of the compound. ND = Not Determined.

[0441] From the above Figure 1 It can be seen from the experimental results in Table 1 that the compound of the present application has good α4β3δ positive allosteric activity, and its activity is significantly better than that of the positive drug allopregnanolone and the positive compound DS-2.

[0442] Test Example 2: Evaluation of the compound's activity in improving sleep

[0443] PCPA sleep deprivation model

[0444] First, electroencephalogram (EEG) / electromyogram (EMG) recording electrodes were surgically implanted in the mouse cerebral cortex. After 5-7 days of postoperative recovery, the mice were placed in a shielded cage to acclimate to the recording environment for 2-3 days. Basal sleep signals were recorded continuously for 12 hours at night using a BIOPAC MP150 polygraph. Subsequently, an insomnia model was established by intraperitoneal injection of 400 mg / kg PCPA (parachlorophenylamine) for 3 consecutive days. Neuroelectric signals were recorded during the night following modeling, and the effectiveness of the model was verified by comparing sleep structure parameters before and after modeling. Mice with successful modeling were randomly divided into a control group and an experimental group. The positive control drug and the test drug were intraperitoneally injected at the corresponding dose. EEG / EMG signals were recorded overnight under the same conditions for 12 hours after drug administration. The sleep-promoting efficacy of the test compound was evaluated by statistical analysis of sleep latency and the proportion of non-rapid eye movement sleep to rapid eye movement sleep between the two groups.

[0445] The activity results of the representative compound WH20240720 on PCPA are shown in Figure 2 shown.

[0446] Depend on Figure 2 The experimental results show that the representative compound WH20240720 can significantly improve the sleep condition of PCPA insomnia mice.

[0447] Test Example 3: Evaluation of Compounds for Improving Depressive-Like Behaviors

[0448] Acute tail suspension test

[0449] This test example mainly uses the acute tail suspension test to evaluate the improving effect of the compound on the depressive-like behavior of mice. The acute tail suspension test in mice induces a helpless state by hanging the animal's tail to evaluate depressive-like behavior. The experiment uses a homemade inverted L-shaped tail suspension device (50 cm high and 10 cm horizontally extended). The mouse (weighing 20-25 g) is fixed to the device beam at 1 / 3 of the tail end using medical tape, so that its head is suspended vertically and 15 cm from the ground. The experiment was carried out in a quiet, low-light environment (50 lux) and was suspended for 6 minutes. The first 2 minutes were the adaptation period, and the mouse's immobility time (defined as no voluntary movement of the whole body) was recorded by the top camera for the next 4 minutes.

[0450] The results of acute tail suspension test after single and multiple administration of representative compound WH20240720 are shown in Figure 2. Figure 3 shown.

[0451] Depend on Figure 3 The experimental results show that the representative compound WH20240720 can help improve the depressive-like behavior of mice regardless of whether it is administered multiple times or once.

Claims

1. A thiazolopyrimidinone compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the structural formula of the thiazolopyrimidinone compound is shown in formula (I): in, R1 is selected from substituted or unsubstituted aryl, heteroaryl or saturated heterocyclic group; when substituted, the aryl, heteroaryl or saturated heterocyclic group is substituted by a substituent selected from halogen, C1-C6 alkyl, C1-C6 alkoxy, hydroxy, trifluoromethyl or trifluoromethoxy; the heteroaryl or saturated heterocyclic group contains 1, 2, 3 or 4 heteroatoms selected from N, O or S; R2 is selected from substituted or unsubstituted C1-C6 alkyl, C3-C6 cycloalkyl, 6-10 membered aryl, 5-10 membered heteroaryl; when substituted, the alkyl, cycloalkyl, aryl or heteroaryl is substituted with a substituent selected from halogen, C1-C6 alkyl, C1-C6 alkoxy, cyano or nitro; the heteroaryl contains 1 or 2 heteroatoms selected from N, O or S; R3 is selected from substituted or unsubstituted 6-10 membered aryl or 5-10 membered heteroaryl; when substituted, the aryl or heteroaryl is substituted by a substituent selected from halogen, C1-C6 alkyl, C1-C6 alkoxy, cyano or nitro; the heteroaryl contains 1, 2, 3 or 4 heteroatoms selected from N, O or S.

2. The thiazolopyrimidinone compound, its stereoisomer or a pharmaceutically acceptable salt thereof according to claim 1, wherein: R1 is selected from substituted or unsubstituted phenyl, substituted or unsubstituted 5-6 membered heteroaryl or unsubstituted 4-6 membered saturated heterocyclyl; when substituted, the phenyl or heteroaryl is substituted with a substituent selected from fluorine, chlorine, bromine, C1-C6 alkyl, C1-C6 alkoxy, hydroxy, trifluoromethyl or trifluoromethoxy; R2 is selected from C1-C6 alkyl, C3-C6 cycloalkyl, substituted or unsubstituted 6-10 membered aryl, 5-6 membered heteroaryl; when substituted, the aryl or heteroaryl is substituted with a substituent selected from fluorine, chlorine, bromine, C1-C6 alkyl, C1-C6 alkoxy, cyano or nitro; R3 is selected from substituted or unsubstituted 6-10 membered aryl or 5-10 membered heteroaryl; when substituted, the aryl or heteroaryl is substituted by a substituent selected from fluorine, chlorine, bromine, C1-C6 alkyl, C1-C6 alkoxy, cyano, and nitro.

3. The thiazolopyrimidinone compound according to claim 1 or 2, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R1 is selected from a 6-membered saturated heterocyclic group, a substituted or unsubstituted phenyl group, or a 5-6-membered heteroaryl group; when substituted, the phenyl group or heteroaryl group is substituted by a substituent selected from fluorine, chlorine, bromine, methyl, vinyl, n-propyl, isopropyl, tert-butyl, methoxy, ethoxy, hydroxy, trifluoromethyl, or trifluoromethoxy; the heteroaryl group or saturated heterocyclic group contains 1, 2, or 3 heteroatoms selected from N, O, or S; R2 is selected from C1-C4 alkyl, C3-C6 cycloalkyl, substituted or unsubstituted phenyl or 6-membered heteroaryl; when substituted, the phenyl or heteroaryl is substituted by a substituent selected from fluorine, chlorine, bromine, C1-C6 alkyl or C1-C6 alkoxy; the heteroaryl contains 1, 2 or 3 heteroatoms selected from N, O or S; R3 is selected from substituted or unsubstituted phenyl or 6-membered heteroaryl; when substituted, the phenyl or heteroaryl is substituted by a substituent selected from fluorine, chlorine, bromine, C1-C6 alkyl, C1-C6 alkoxy or nitro; the heteroaryl contains 1, 2 or 3 heteroatoms selected from N, O or S.

4. The thiazolopyrimidinone compound according to any one of claims 1 to 3, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: In R1, the heteroaryl group contains 1 or 2 heteroatoms selected from N or S; In R2, the heteroaryl group contains 1 or 2 N atoms; In R3, the heteroaryl group contains 1 or 2 N atoms.

5. The thiazolopyrimidinone compound according to any one of claims 1 to 4, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R1 is selected from morpholinyl, substituted or unsubstituted phenyl or 5-6 membered heteroaryl, wherein the heteroaryl is selected from pyrimidinyl, pyridinyl, thienyl, furanyl, pyrrolyl or pyrazolyl; when substituted, the phenyl or heteroaryl is substituted with a substituent selected from fluorine, chlorine, methyl, methoxy, hydroxy or trifluoromethyl; R2 is selected from methyl, ethyl, isopropyl, cyclopropyl, substituted or unsubstituted phenyl, pyrimidinyl or pyrimidinyl; when substituted, the phenyl, pyrimidinyl or pyrimidinyl is substituted with a substituent selected from fluoro, chloro or methyl; R3 is selected from substituted or unsubstituted phenyl, pyridyl or pyrimidinyl; when substituted, the phenyl, pyridyl or pyrimidinyl is substituted by a substituent selected from fluorine, chlorine, methyl or nitro.

6. The thiazolopyrimidinone compound according to any one of claims 1 to 5, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R1 is selected from morpholinyl, thienyl, pyrimidinyl, methoxypyrimidinyl, hydroxyphenyl, fluorophenyl, methoxyphenyl, pyridyl, imidazolyl; R2 is selected from methyl, ethyl, cyclopropyl or phenyl; R3 is selected from pyridyl substituted by methyl, substituted or unsubstituted phenyl; when substituted, the phenyl is substituted by 1 or 2 substituents selected from methyl, ethyl, fluorine or chlorine.

7. The thiazolopyrimidinone compound according to any one of claims 1 to 6, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R1 is selected from thienyl or pyrimidinyl substituted by methoxy; R2 is selected from methyl, ethyl, cyclopropyl or unsubstituted phenyl; R3 is selected from phenyl groups substituted by methyl, ethyl, fluorine or chlorine, and the number of substituents of the phenyl group is 1 or 2.

8. The thiazolopyrimidinone compound according to any one of claims 1 to 7, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: The thiazolopyrimidinone compound is selected from one of the following structures:

9. A pharmaceutical composition comprising the thiazolopyrimidinone compound according to any one of claims 1 to 8, its stereoisomers or pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier; Preferably, the pharmaceutical composition is a gastrointestinal dosage form or an injectable dosage form; more preferably, the gastrointestinal dosage form is selected from powders, tablets, granules, capsules, solutions, emulsions or suspensions; more preferably, the injectable dosage form is selected from intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection or intracavitary injection.

10. Use of the thiazolopyrimidinone compound according to any one of claims 1 to 8, its stereoisomer or pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 9 in the preparation of a drug for improving sleep or treating depression; Preferably, the thiazolopyrimidinone compound, its stereoisomer or pharmaceutically acceptable salt thereof is used as GABA A Receptor modulators.

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