New deuterium-containing pyrazolone compound, and pharmaceutical composition and use thereof

Abstract

Disclosed are a new deuterium-containing pyrazolone compound, and a pharmaceutical composition and the use thereof. The new deuterium-containing pyrazolone compound is as represented by formula (I) and / or formula (II), wherein the definition of each substituent is detailed in the description. The compound can be used in the preparation of a drug for preventing or treating neurodegenerative diseases and cardiovascular and cerebrovascular diseases.

Description

A novel deuterium-containing pyrazolone compound, its pharmaceutical composition, and its uses. Technical Field

[0001] This invention relates to, but is not limited to, the field of pharmaceutical chemistry, and particularly to a novel deuterium-containing pyrazolinone compound, tautomer, stereoisomer, prodrug, pharmaceutically acceptable salt thereof, pharmaceutical composition thereof, and uses thereof. Background Technology

[0002] Neurodegenerative diseases (NDDs) are a group of neurological disorders characterized by the progressive loss of neurons in the central nervous system (CNS) or peripheral nervous system (PNS). These chronic, progressive neurological illnesses affect the lives of millions worldwide. Since 1990, the total number of disabilities, illnesses, and premature deaths (disability-adjusted life years) caused by neurological diseases has increased by 18%. These diseases primarily include Alzheimer's disease, Parkinson's disease, Huntington's disease, various types of spinocerebellar ataxia, multiple sclerosis, cerebellar atrophy, and amyotrophic lateral sclerosis (ALS). Research has found that neurodegenerative diseases are caused by a variety of factors, including insufficient nutrition to neurons or glial cells, excessive glutamate receptor activity, high levels of reactive oxygen species, impaired metabolic pathways, reduced mitochondrial energy production, inflammation, viral infections, and mutations in nuclear or mitochondrial DNA. These factors interact with each other, ultimately leading to neurological dysfunction and cell death. Due to the complex and diverse mechanisms of action, there are currently no effective and mature methods or drugs to prevent or treat these diseases. Therefore, finding a highly effective and multi-targeted drug has significant social and economic value. Summary of the Invention

[0003] The inventors have developed a novel deuterium-containing pyrazolone compound that exhibits better protection against L-glutamate-induced PC12 cell damage, better improvement in ischemic stroke, easier crossing of the blood-brain barrier, and longer retention time in brain tissue and cerebrospinal fluid. Unexpectedly, it has a short half-life, which effectively avoids accumulation in vivo and reduces hepatotoxic side effects.

[0004] In one aspect, this invention provides a novel deuterium-containing pyrazolinone compound, tautomer, stereoisomer, prodrug, and pharmaceutically acceptable salt thereof, as shown in (I) and / or (II):

[0005] In formula (I) and / or formula (II),

[0006] R1 is selected from hydrogen, deuterium, C1-C8 alkyl group substituted or unsubstituted with one or more substituents, phenyl group substituted or unsubstituted with one or more substituents, or pyridyl group substituted or unsubstituted with one or more substituents, wherein the substituents are selected from deuterium or halogens;

[0007] R2 is selected from hydrogen, deuterium, C1-C8 alkyl group substituted or unsubstituted with one or more substituents, phenyl group substituted or unsubstituted with one or more substituents, or pyridyl group substituted or unsubstituted with one or more substituents, wherein the substituents are selected from deuterium or halogens;

[0008] R3 is selected from hydrogen or deuterium;

[0009] R4 is selected from cyclohexyl groups substituted or unsubstituted with one or more substituents, benzyl groups substituted or unsubstituted with one or more substituents, naphthyl groups substituted or unsubstituted with one or more substituents, heterocyclic groups substituted or unsubstituted with one or more substituents, or... in,

[0010] The above substituents are selected from deuterium or halogens;

[0011] R X1 Selected from halogens, or C1-C8 alkyl groups substituted or unsubstituted by one or more substituents, wherein the substituents are selected from deuterium or halogens;

[0012] R X2 Selected from halogens, or C1-C8 alkyl groups substituted or unsubstituted by one or more substituents, wherein the substituents are selected from deuterium or halogens;

[0013] R X3 Selected from halogens, C1-C8 alkyl groups substituted or unsubstituted with one or more substituents, and C1-C8 alkoxy groups substituted or unsubstituted with one or more substituents. in,

[0014] The substituents mentioned above are selected from deuterium and hydroxyl groups;

[0015] The above R X4 Selected from C1-C8 alkyl groups substituted or unsubstituted by one or more substituents, or C1-C8 alkoxy groups substituted or unsubstituted by one or more substituents, wherein the substituents are selected from deuterium or halogens;

[0016] In particular,

[0017] At least one of R1, R2, R3, and R4 is either deuterium or is replaced by deuterium.

[0018] In some embodiments, the present invention provides a novel deuterium-containing pyrazolinone compound, tautomer, stereoisomer, prodrug, and pharmaceutically acceptable salt thereof as shown in formula (III) and / or formula (IV):

[0019] The substituents in formula (III) and / or formula (IV) are defined as defined in formula (I) and / or formula (II) above.

[0020] In some embodiments, in formulas (I)-(IV) above, R1 is selected from hydrogen, deuterium, C1-C8 alkyl substituted or unsubstituted with one or more substituents, phenyl substituted or unsubstituted with one or more substituents, or pyridyl substituted or unsubstituted with one or more substituents, wherein the substituents are selected from deuterium or halogens; preferably, R1 is selected from methyl, trideuterated methyl, phenyl, or propyl.

[0021] In some embodiments, in formulas (I)-(IV) above, R2 is selected from hydrogen, deuterium, C1-C8 alkyl substituted or unsubstituted with one or more substituents, phenyl substituted or unsubstituted with one or more substituents, or pyridyl substituted or unsubstituted with one or more substituents, wherein the substituents are selected from deuterium or halogens; preferably, R2 is selected from hydrogen, deuterium, or isobutyl.

[0022] In some implementations, in formulas (I) and / or (III) above, R3 is selected from hydrogen or deuterium;

[0023] In some embodiments, in formulas (I) and / or (II) above, R4 is selected from cyclohexyl groups substituted or unsubstituted with one or more substituents, benzyl groups substituted or unsubstituted with one or more substituents, naphthyl groups substituted or unsubstituted with one or more substituents, heterocyclic groups substituted or unsubstituted with one or more substituents, or... Wherein, the substituents are selected from deuterium or halogens; preferably, R4 is selected from cyclohexyl, benzyl, naphthyl, pyridyl, benzothiophene, or in,

[0024] The above R X1 Selected from halogens, or C1-C8 alkyl groups substituted or unsubstituted with one or more substituents, wherein the substituents are selected from deuterium or halogens; preferably, R X1 Selected from chloro, methyl, and trideuterated methyl;

[0025] R X2 Selected from halogens, or C1-C8 alkyl groups substituted or unsubstituted with one or more substituents, wherein the substituents are selected from deuterium or halogens; preferably, R X2 Selected from methyl and trideuterated methyl;

[0026] R X3 Selected from halogens, C1-C8 alkyl groups substituted or unsubstituted with one or more substituents, and C1-C8 alkoxy groups substituted or unsubstituted with one or more substituents. in,

[0027] The substituents mentioned above are selected from deuterium and hydroxyl groups;

[0028] The above R X4Selected from C1-C8 alkyl groups substituted or unsubstituted by one or more substituents, or C1-C8 alkoxy groups substituted or unsubstituted by one or more substituents, wherein the substituents are selected from deuterium or halogens;

[0029] Preferably, R X3 Selected from chlorine, methoxy, ethoxy, hydroxymethyl, -CDHOH, -CD2OH, ethoxyacyl, or

[0030] In some implementations, R4 in equations (I)-(IV) above is selected from... in,

[0031] The above R X1 Selected from halogens, or C1-C8 alkyl groups substituted or unsubstituted with one or more substituents, wherein the substituents are selected from deuterium or halogens; preferably, R X1 Selected from chloro, methyl, and trideuterated methyl;

[0032] R X2 Selected from halogens, or C1-C8 alkyl groups substituted or unsubstituted with one or more substituents, wherein the substituents are selected from deuterium or halogens; preferably, R X2 Selected from methyl and trideuterated methyl;

[0033] R X3 Selected from halogens, C1-C8 alkyl groups substituted or unsubstituted with one or more substituents, and C1-C8 alkoxy groups substituted or unsubstituted with one or more substituents. in,

[0034] The substituents mentioned above are selected from deuterium and hydroxyl groups;

[0035] The above R X4 Selected from C1-C8 alkyl groups substituted or unsubstituted by one or more substituents, or C1-C8 alkoxy groups substituted or unsubstituted by one or more substituents, wherein the substituents are selected from deuterium or halogens;

[0036] Preferably, R X3 Selected from chlorine, methoxy, ethoxy, hydroxymethyl, -CDHOH, -CD2OH, ethoxyacyl, or

[0037] In particular,

[0038] At least one of R1, R2, R3, and R4 is either deuterium or is replaced by deuterium.

[0039] In some embodiments, the novel deuterium-containing pyrazolone compounds, tautomers, stereoisomers, prodrugs, and pharmaceutically acceptable salts provided by the present invention are selected from the following compounds:

[0040] On the other hand, the present invention provides a pharmaceutical composition comprising the above-mentioned novel deuterium-containing pyrazolone compound, tautomer, stereoisomer, prodrug, and pharmaceutically acceptable salt thereof.

[0041] This invention discloses a pharmaceutical composition comprising the compounds, tautomers, stereoisomers, prodrugs and their pharmaceutically acceptable salts described in this invention as active ingredients or main active ingredients, supplemented by a pharmaceutically acceptable carrier.

[0042] In another aspect, the present invention provides the use of the above-mentioned novel deuterium-containing pyrazolone compounds, tautomers, stereoisomers, prodrugs and their pharmaceutically acceptable salts or the above-mentioned pharmaceutical compositions in the preparation of neuroprotective drugs.

[0043] Thirdly, the present invention provides the use of the above-mentioned novel deuterium-containing pyrazolone compounds, tautomers, stereoisomers, prodrugs and their pharmaceutically acceptable salts or the above-mentioned pharmaceutical compositions in the preparation of medicaments for the prevention or treatment of cardiovascular and cerebrovascular diseases.

[0044] This invention provides the use of the above-mentioned pharmaceutical composition in the preparation of neuroprotective drugs, wherein the neuroprotective drugs are drugs for treating neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, Huntington's disease, multiple sclerosis, cerebellar ataxia, different types of spinocerebellar ataxia, spinal muscular atrophy, cerebral ischemia, and primary lateral sclerosis.

[0045] This invention provides the use of the above-mentioned pharmaceutical composition in the preparation of drugs for the prevention or treatment of cardiovascular and cerebrovascular diseases, wherein the drugs for the prevention or treatment of cardiovascular and cerebrovascular diseases are drugs for the treatment of cardiovascular and cerebrovascular diseases, including hypertension, coronary heart disease, stroke, diabetic heart failure, heart failure, diastolic heart failure, systolic heart failure, postoperative volume overload, idiopathic edema, pulmonary hypertension, pulmonary arterial hypertension, acute decompensated heart failure, heart failure, acute renal failure, and nephrotic syndrome.

[0046] In some embodiments, the novel compounds of the present invention can be formulated as pharmaceutical compositions and administered to patients via a variety of suitable routes of administration, including systemic (e.g., oral or parenteral), intravenous, intramuscular, transdermal, or subcutaneous routes.

[0047] The inventors have developed a novel deuterium-containing pyrazolone compound that exhibits better protection against L-glutamate-induced PC12 cell damage, better improvement in ischemic stroke, easier crossing of the blood-brain barrier, and longer retention time in brain tissue and cerebrospinal fluid. Unexpectedly, it has a short half-life, which effectively avoids accumulation in vivo and reduces hepatotoxic side effects.

[0048] definition:

[0049] Unless otherwise stated, the following terms and phrases as used herein are intended to have the following meanings. A particular term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient.

[0050] Some compounds of this invention can exist in either a solvated or a solvent-based form, such as hydrates or ethanolates. Generally, the solvent-based and the solvent-based forms are equivalent and are both included within the scope of this invention.

[0051] The term "pharmaceutical acceptable" refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0052] The term "pharmaceutically acceptable salt" refers to a salt of the compounds of this invention, prepared by reacting a compound with a relatively non-toxic acid or base, as discovered in this invention, with a specific substituent. When the compounds of this invention contain relatively acidic functional groups, base addition salts can be obtained by contacting a neutral form of such compound with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include aluminum, sodium, potassium, calcium, manganese, iron, ammonium, organic amine, or magnesium salts, or similar salts. When the compounds of this invention contain relatively basic functional groups, acid addition salts can be obtained by contacting a neutral form of such compound with a sufficient amount of acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts, such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and organic acid salts, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, octanoic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; as well as salts of amino acids (such as arginine) and salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain both basic and acidic functional groups, and thus can be converted into either a base or an acid addition salt.

[0053] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, including straight-chain and branched groups. Alkyl groups can be substituted or unsubstituted. When substituted, the substituent is preferably one or more, more preferably one to three, and most preferably one or two.

[0054] The term "alkenyl" refers to an aliphatic hydrocarbon group containing an unsaturated carbon-carbon double bond, including straight-chain and branched groups. The alkyl group can be substituted or unsubstituted. There can be one or more carbon-carbon double bonds.

[0055] The term "cycloalkyl" refers to a monocyclic or fused-ring group consisting entirely of carbon atoms (a "fused" ring means that each ring in the system shares a pair of adjacent carbon atoms with the other rings in the system), wherein one or more rings do not have a fully connected π-electron system. Examples of cycloalkyl groups (but not limited to) include cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, adamantane, cyclohexadiene, cycloheptane, and cyclohepttriene. Cycloalkyl groups can be substituted or unsubstituted.

[0056] The term "aryl" refers to an all-carbon monocyclic or fused polycyclic group with 1 to 12 carbon atoms and a fully conjugated π-electron system. Non-limiting examples of aryl groups include phenyl, naphthyl, and anthracene. Aryl groups can be substituted or unsubstituted. When substituted, the substituents are preferably one or more, more preferably one, two, or three, and even more preferably one or two.

[0057] The term "aryl hydrocarbon group" refers to a hydrocarbon group that has been replaced by an aryl group.

[0058] The term "heteroaryl" refers to a monocyclic or fused cyclic group of multiple atoms containing one, two, three, or four cyclic heteroatoms selected from N, O, or S, with the remaining cyclic atoms being C, and possessing a fully conjugated π-electron system. Non-limiting examples of unsubstituted heteroaryl groups include pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyrimidine, quinoline, isoquinoline, purine, tetrazolium, triazine, and carbazole.

[0059] The term "alkoxy" refers to an alkyl group bonded to an oxygen atom, where the alkyl group can be straight-chain, branched, or cycloalkyl.

[0060] The term "hydroxyl group" refers to the -OH group.

[0061] The term "amino" refers to the -NH2 group.

[0062] The term "carboxyl group" refers to the -COOH group.

[0063] The term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0064] The term "pharmaceutically acceptable carrier" refers to any formulation or carrier medium capable of delivering an effective amount of the active substance of this invention without interfering with the biological activity of the active substance and without toxic side effects on the host or patient. Representative carriers include water, oil, vegetables and minerals, ointment bases, lotion bases, and ointment bases. These bases include suspending agents, thickeners, transdermal penetration enhancers, etc.

[0065] The term "stereoisomer" refers to compounds that have the same chemical composition but different spatial arrangements of atoms or groups.

[0066] The numerical range mentioned in this application, such as "C1-C8", means that the group can contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to 8 carbon atoms. Detailed Implementation

[0067] The following examples provide numerous exemplary methods for preparing the compounds of the present invention. The invention is described in detail below through examples, but this does not imply any adverse limitation thereof. The invention has been described in detail herein, and specific embodiments thereof are also disclosed. It will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the invention without departing from the spirit and scope thereof. Some compounds of the present invention can be used as intermediates for preparing other compounds of the present invention; the structures of all compounds have been determined by liquid chromatography-mass spectrometry.

[0068] Unless otherwise specified, all materials used in the embodiments of this application were purchased commercially.

[0069] Example 1: Synthesis of compound ZJT1

[0070] Reaction formula:

[0071] Preparation method:

[0072] Under nitrogen atmosphere, 3.5 g (28.6 mmol) of 2,3,5-trimethylpyrazine was added to 35 mL of heavy water, followed by 5 mL of 40% deuterated sodium hydroxide solution, and refluxed for five days. Liquid chromatography-mass spectrometry (LC-MS) analysis showed the reaction was complete.

[0073] The reaction mixture was extracted with dichloromethane (DCM, 50 mL × 3), the organic phases were combined, dried over anhydrous magnesium sulfate, filtered, and concentrated to give 3.0 g of liquid (yield: 79.9%). ESI-MS(+): m / z = 132.07 [M+1].

[0074] Example 2: Synthesis of compound ZJT2

[0075] Reaction formula:

[0076] Preparation method:

[0077] Step 1: Preparation of compound ZJT-2-1

[0078] ZJT-2-SM (880 mg, 10 mmol, 1 eq) was cooled to -78 °C, and then bis(trimethylsilyl)aminolithium (1.0 mol / L tetrahydrofuran) (11 mL, 11 mmol, 1.1 eq) was added, and the reaction was continued for 10 min. Then acetyl chloride-3d (815 mg, 10 mL, 1.0 eq) was added, and the system was kept at -78 °C, and the reaction was continued for 2 h. Liquid chromatography monitoring showed that the reaction was complete.

[0079] The system was quenched in water (25 mL), and the aqueous phase was extracted with ethyl acetate (30 mL × 3). The organic phases were combined and dried over anhydrous sodium sulfate. The mixture was filtered and concentrated to give 1 g of residue. This residue was not purified and was reserved for further processing. ESI-MS(+): m / z = 134.13 [M+1].

[0080] Step 2: Preparation of compound ZJT-2

[0081] ZJT-2-1 (1g) was added to hydrazine hydrate (10ml), and the system was heated to 120℃ and reacted for 3 hours. Thin-layer chromatography showed that the reaction was complete. The system was cooled to room temperature and concentrated to obtain the crude product.

[0082] The crude product was subjected to column chromatography (petroleum ether: ethyl acetate = 30:1) to give 653 mg of solid (yield: 86.1%). ESI-MS(+): m / z = 102.09 [M+1].

[0083] Example 3: Synthesis of compound ZJT3

[0084] Reaction formula:

[0085] Preparation method:

[0086] Step 1: Preparation of compound ZJT-3-1

[0087] Add Michaelis acid (2.88 g, 20 mmol, 1 eq) to heavy water (30 ml), then add potassium carbonate (5.52 g, 40 mmol, 2 eq), and continue the reaction for 1 hour. Liquid chromatography monitoring showed the reaction was complete.

[0088] Filter, concentrate to dryness, dissolve the residue in ethyl acetate (60 mL), and dry with anhydrous sodium sulfate. Filter, concentrate, and give 2.84 g of the target substance (yield: 97.2%). ESI-MS(+): m / z = 147.23 [M+1].

[0089] Step 2: Preparation of compound ZJT-3-2

[0090] ZJT-3-1 (2.8 g, 19.2 mol, 1 eq) was added to DCM (80 ml), followed by pyridine (1.7 g, 21.1 mmol, 1.1 eq). The system was cooled to -10 °C, and acetyl chloride-3d (1.7 g, 21.1 mmol, 1.1 eq) was added. The reaction was continued for 2 hours. The reaction was monitored by liquid chromatography and found to be complete.

[0091] The system was quenched in water (70 mL), and the aqueous phase was extracted with ethyl acetate (50 mL × 3). The organic phases were combined and dried over anhydrous sodium sulfate. The mixture was filtered and concentrated to give 3.41 g of the target analyte (yield: 93.5%). ESI-MS(+): m / z = 191.13 [M+1].

[0092] Step 3: Preparation of compound ZJT-3-3

[0093] ZJT-3-2 (2.8 g, 20.0 mmol) was added to deuterated ethanol-1d (20 ml), and the system was heated to reflux and reacted for 4 hours. Liquid chromatography-mass monitoring showed that the reaction was complete after the molecular weight of the starting material was no longer present.

[0094] The system was cooled to room temperature and concentrated to obtain 2.4 g of the target compound (yield: 88.9%). ESI-MS(+): m / z = 136.09 [M+1].

[0095] Step 4: Preparation of compound ZJT-3

[0096] ZJT-3-3 (1.35 g, 10 mmol) was added to hydrazine hydrate (15 ml), and the system was heated to 120 °C and reacted for 1 hour. Thin-layer chromatography showed that the reaction was complete. The system was cooled to room temperature and concentrated to give the crude product. The crude product was subjected to column chromatography (petroleum ether: ethyl acetate = 30:1) to give 800 mg of white solid (yield: 77.6%). ESI-MS(+): m / z = 104.19 [M+1].

[0097] Example 4: Synthesis of compound DSC207-01

[0098] Reaction formula:

[0099] Preparation method:

[0100] Step 1: Preparation of compound DSC207-01-1

[0101] ZJT-1 (25.0 g, 190.5 mmol, 1.0 eq) and (5 g, 38.1 mmol, 1 eq) were added to DCM (250 ml). The system was cooled to -10 °C, and m-chloroperoxybenzoic acid (m-CPBA, 36.2 g, 209.5 mmol, 1.1 eq) was added in portions. After the addition was complete, the reaction was allowed to proceed for another 40 min.

[0102] Thin-layer chromatography showed that the reaction was complete. The system was heated to 25°C and poured into an aqueous sodium sulfite solution (250 mL). The mixture was separated, and the aqueous phase was extracted with DCM (150 mL × 3). The organic phases were combined, dried over sodium sulfate, and concentrated to give the target compound DSC207-01-125.0 g (yield: 89.2%). ESI-MS(+): m / z = 148.89 [M+1].

[0103] Step 2: Preparation of compound DSC207-01-2

[0104] DSC207-01-1 (25.0 g, 170.0 mmol, 1 eq) was added to toluene (250 ml), the system was cooled to -10 °C, and then phosphorus oxychloride (31.3 g, 20.4 mmol, 1.2 eq) was added. After the addition was complete, the system was heated to 90 °C and reacted for 16 hours.

[0105] Thin-layer chromatography showed the reaction was complete. The mixture was concentrated, and the residue was dissolved in DCM (100 ml). The pH was adjusted to 8-9 with sodium bicarbonate aqueous solution. The mixture was separated, and the aqueous phase was extracted with DCM (150 ml × 3). The organic phases were combined, washed once with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give the crude product. The crude product was subjected to column chromatography (petroleum ether: ethyl acetate = 20:1) to give 13.5 g of solid (yield: 47.9%). ESI-MS(+): m / z = 166.19 [M+1].

[0106] Step 3: Preparation of compound DSC207-01-3

[0107] DSC207-01-2 (10 g, 60.4 mmol) was added to hydrazine hydrate (100 ml), and the system was heated to 120 °C and reacted for 6 hours. Thin-layer chromatography showed that the reaction was complete. The system was cooled to room temperature and concentrated to obtain the crude product. Column chromatography of the crude product (dichloromethane:methanol = 50:1) gave 8.1 g of a white solid.

[0108] (Yield: 83.2%). ESI-MS(+): m / z = 162.01 [M+1].

[0109] Step 4: Preparation of compound DSC207-01-4

[0110] DSC207-01-3 (4.0 g, 24.8 mmol, 1 eq) was added to acetic acid (30 mL), followed by ethyl acetoacetate (3.54 g, 27.28 mmol, 1.1 eq). The system was heated to 90 °C and reacted for 1.5 h. Thin-layer chromatography showed that the reaction was complete. The system was cooled to room temperature and concentrated to give the crude product. Column chromatography of the crude product (petroleum ether: ethyl acetate = 10:1) gave 3.7 g of a yellow solid (yield: 65.6%).

[0111] ESI-MS(+): m / z=228.79[M+1].

[0112] Step 5: Preparation of compound DSC207-01-5

[0113] DSC207-01-4 (7.0 g, 30.8 mmol, 1 eq) was added to acetic anhydride (70 mL), and the system was heated to 130 °C and reacted for 3 hours. Thin-layer chromatography showed that the reaction was complete. The system was cooled to room temperature and concentrated to give the crude product. The crude product was subjected to column chromatography (petroleum ether: ethyl acetate = 20:1) to give 6.4 g of oil (yield: 77.1%). ESI-MS(+): m / z = 270.39 [M+1].

[0114] Step 6: Preparation of compound DSC207-01-6

[0115] DSC207-01-5 (4.0 g, 14.9 mmol, 1 eq) was added to DCM (40 mL), and the system was cooled to -10 °C. m-CPBA (5.13 g, 29.7 mmol, 2 eq) was added in portions, and the reaction was allowed to proceed for another 30 min after each addition. Thin-layer chromatography showed that the reaction was complete. The system was heated to 25 °C and poured into a sodium sulfite aqueous solution (50 mL). The mixture was separated, and the aqueous phase was extracted with DCM (50 mL × 3). The organic phases were combined, dried over sodium sulfate, and concentrated to obtain the crude product. The crude product was subjected to column chromatography (petroleum ether: ethyl acetate = 10:1) to give 3.5 g of solid (yield: 82.5%). ESI-MS(+): m / z = 286.29 [M+1].

[0116] Step 7: Preparation of compound DSC207-01-7

[0117] DSC207-01-6 (3.0 g, 10.5 mmol, 1 eq) was added to acetic anhydride (30 mL), and the system was heated to 130 °C and reacted for 5 hours. Thin-layer chromatography showed that the reaction was complete. The system was cooled to room temperature and concentrated to obtain the crude product. The crude product was subjected to column chromatography (petroleum ether: ethyl acetate = 10:1) to give 1.9 g of the target substance (yield: 55.8%). ESI-MS(+): m / z = 327.89 [M+1].

[0118] Step 8: Preparation of compound DSC207-01

[0119] Add DSC207-01-7 (1.5 g, 4.6 mmol, 1 eq) to a 25 mL single-necked flask, then add methanol (15 mL) and lithium hydroxide (220 mg, 9.2 mmol, 2.5 eq). React at room temperature for 16 hours. TLC detection showed the starting material had disappeared. Concentrate the mixture, add water (15 mL) to the residue, adjust the pH to 6-7 with 4N hydrochloric acid, extract the aqueous phase with DCM (20 mL × 3), combine the organic phases, wash once with saturated brine, dry over anhydrous sodium sulfate, concentrate, and obtain the crude product.

[0120] The crude sample was subjected to column chromatography (dichloromethane:methanol = 30:1) to obtain 640 mg of the target analyte (yield: 57.5%). ESI-MS(+): m / z = 243.14 [M+1].

[0121] Example 5: Synthesis of compound DSC207-03

[0122] Reaction formula:

[0123] Preparation method:

[0124] Step 1: Preparation of compound DSC207-03-1

[0125] Compound DSC207-03-SM (8.0 g, 52.6 mmol) was dissolved in methanol (80 mL). The system was cooled to 0 °C, and 2 drops of sulfuric acid were added. The system was then allowed to react at room temperature for 3 hours. Thin-layer chromatography showed that the reaction was essentially complete. The system was concentrated, and the residue was poured into water and extracted with ethyl acetate (100 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give 7.0 g of a yellow oil (yield: 80.1%).

[0126] ESI-MS(+): m / z=167.18[M+1].

[0127] Step 2: Preparation of compound DSC207-03-2

[0128] DSC207-03-1 (7.0 g, 42.1 mmol, 1 eq) was added to DCM (70 mL), and the system was cooled to -10 °C. m-CPBA (8.7 g, 50.5 mmol, 1.2 eq) was added in portions, and the reaction was allowed to proceed at room temperature for 8 hours after the addition was complete. Thin-layer chromatography showed that the reaction was complete. The system was heated to 25 °C and poured into an aqueous sodium sulfite solution (50 mL). The mixture was separated, and the aqueous phase was extracted with DCM (50 mL × 3). The organic phases were combined, dried over sodium sulfate, and concentrated to give 7.0 g of a white solid (yield: 91.3%).

[0129] ESI-MS(+): m / z=183.59[M+1].

[0130] Step 3: Preparation of compound DSC207-03-3

[0131] DSC207-03-2 (5.0 g, 27.4 mmol, 1 eq) was added to toluene (25 mL), and the system was cooled to -10 °C. Phosphorus oxychloride (5.0 g, 32.9 mmol, 1.2 eq) was then added. After the addition was complete, the system was heated to 90 °C and reacted for 16 hours. Thin-layer chromatography showed that the reaction was complete. The mixture was concentrated, and the residue was dissolved in DCM (20 mL). The pH was adjusted to 8-9 with sodium bicarbonate aqueous solution. The mixture was separated, and the aqueous phase was extracted with DCM (50 mL × 3). The organic phases were combined, washed once with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was subjected to column chromatography (petroleum ether: ethyl acetate = 20:1) to obtain 4.7 g of the target analyte (yield 85.4%). ESI-MS (+): m / z = 201.09 [M+1].

[0132] Step 4: Preparation of compound DSC207-03-4

[0133] 3-Methyl-2-pyrazolin-5-one (1.6 g, 14.0 mmol, 1.4 eq) was added to dimethyl sulfoxide (DMSO, 30 mL), and the system was cooled to -10 °C. Potassium carbonate (2.78 g, 19.9 mmol, 2 eq) was then added. After the addition was complete, the mixture was stirred at room temperature for ten minutes. DSC207-03-3 (2.0 g, 10.0 mmol, 1 eq) was then added, and the mixture was allowed to react at room temperature for 3 hours. Thin-layer chromatography showed that the reaction was complete. The system was poured into water, the pH was adjusted to 3-4 with 1N hydrochloric acid, and the mixture was extracted with ethyl acetate (15 mL × 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product.

[0134] The crude sample was subjected to column chromatography (petroleum ether: ethyl acetate = 10:1) to give 2.0 g of a white solid (yield: 76.9%). ESI-MS(+): m / z = 263.29 [M+1].

[0135] Step 5: Preparation of compound DSC207-03

[0136] DSC207-03-4 (1.0 g, 3.8 mmol, 1 eq) was added to tetrahydrofuran (THF, 10 mL), and the system was cooled to -10 °C. Lithium aluminum hydride-4d (1 mol / L in THF, 4.19 mL, 4.19 mmol, 1.1 eq) was added, and the mixture was allowed to react at room temperature for 4 hours. Thin-layer chromatography showed that the reaction was complete. The system was poured into an aqueous solution (50 mL), and the aqueous phase was extracted with DCM (50 mL × 3). The organic phases were combined, dried over sodium sulfate, and concentrated to give 720 mg of an oil (yield: 80.0%). ESI-MS(+): m / z = 237.29 [M+1].

[0137] Example 6: Synthesis of compound DSC207-04

[0138] Reaction formula:

[0139] Preparation method:

[0140] DSC207-03-4 (1.0 g, 3.8 mmol, 1 eq) was added to methanol (10 mL), the system was cooled to -10 °C, sodium borohydride-4d (0.64 g, 15.2 mmol, 4 eq) was added, and after the addition was complete, the mixture was allowed to rise to room temperature and react for 4 hours.

[0141] Thin-layer chromatography showed that the reaction was complete. The system was poured into an aqueous solution (50 mL), and the aqueous phase was extracted with DCM (50 mL × 3). The organic phases were combined, dried over sodium sulfate, concentrated, and purified by column chromatography to give 494 mg of the target substance (yield: 55.3%). ESI-MS(+): m / z = 236.49 [M+1].

[0142] Example 7: Synthesis of compound DSC207-05

[0143] Reaction formula:

[0144] Preparation method:

[0145] Step 1: Preparation of compound DSC207-05-1

[0146] ZJT-2 (1.47 g, 14.5 mmol, 1.45 eq) was added to DMSO (30 mL), the system was cooled to -10 °C, and then potassium carbonate (2.75 g, 20.0 mmol, 2 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 10 minutes, and then DSC207-03-3 (2.0 g, 10.0 mmol, 1 eq) was added. After the addition was complete, the mixture was reacted at room temperature for 3 hours.

[0147] Thin-layer chromatography showed the reaction was complete. The system was poured into water, the pH was adjusted to 3-4 with 1N hydrochloric acid, and the mixture was extracted with ethyl acetate (15ml × 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give the crude product. The crude product was subjected to column chromatography (petroleum ether:ethyl acetate = 10:1) to give 1.3g of the target analyte (yield: 49.0%). ESI-MS(+): m / z = 266.29 [M+1].

[0148] Step 2: Preparation of compound DSC207-05

[0149] DSC207-05-1 (1.0 g, 3.8 mmol, 1 eq) was added to methanol (10 mL), the system was cooled to -10 °C, sodium borohydride-4d (0.64 g, 15.2 mmol, 4 eq) was added, and after the addition was complete, the mixture was allowed to rise to room temperature and react for 4 hours.

[0150] Thin-layer chromatography showed that the reaction was complete. The system was poured into an aqueous solution (50 mL), and the aqueous phase was extracted with DCM (50 mL × 3). The organic phases were combined, dried over sodium sulfate, concentrated, and purified by column chromatography to give 494 mg of the target substance (yield: 55.3%). ESI-MS(+): m / z = 238.23 [M+1].

[0151] Example 8: Synthesis of compound DSC207-06

[0152] Reaction formula:

[0153] Preparation method:

[0154] Step 1: Preparation of compound DSC207-06-3

[0155] ZJT-2 (1.46 g, 14.5 mmol, 1.45 eq) was added to DMSO (30 mL), the system was cooled to -10 °C, and then potassium carbonate (2.75 g, 19.9 mmol, 2 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 10 minutes, and then DSC207-01-2 (1.65 g, 9.96 mmol, 1 eq) was added. After the addition was complete, the mixture was reacted at room temperature for 3 hours.

[0156] Thin-layer chromatography showed the reaction was complete. The system was poured into water, the pH was adjusted to 3-4 with 1N hydrochloric acid, and extracted with ethyl acetate (15ml × 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. 1.5g of the target analyte was purified by column chromatography (yield: 65.5%). ESI-MS(+): m / z = 231.33 [M+1].

[0157] Step 2: Preparation of compound DSC207-06-2

[0158] Add DSC207-06-3 (4.0 g, 17.4 mmol, 1 eq) to DCM (40 ml), cool the system to -10 °C, add m-CPBA (6.0 g, 34.8 mmol, 2 eq) in portions, and react for another 30 min after the addition is complete.

[0159] Thin-layer chromatography showed that the reaction was complete. The system was heated to 25°C and poured into an aqueous sodium sulfite solution (50 mL). The mixture was separated, and the aqueous phase was extracted with DCM (50 mL × 3). The organic phases were combined, dried over sodium sulfate, and concentrated to obtain the crude product. Column chromatography purified the target analyte to 3.1 g (yield: 72.3%). ESI-MS(+): m / z = 247.42 [M+1].

[0160] Step 3: Preparation of compound DSC207-06-1

[0161] DSC207-06-2 (3.0 g, 12.2 mmol, 1 eq) was added to acetic anhydride (30 mL), and the system was heated to 130 °C and reacted for 5 hours. Thin-layer chromatography showed that the reaction was complete. The system was cooled to room temperature and concentrated to obtain the crude product. The crude product was purified by column chromatography to obtain 2.0 g of the target substance (yield: 57.3%).

[0162] ESI-MS(+): m / z=288.45[M+1].

[0163] Step 4: Preparation of compound DSC207-06

[0164] Add starting material DSC207-06-1 (1.32 g, 4.6 mmol, 1 eq) to a 50 mL single-necked flask, then add methanol (15 mL) and lithium hydroxide (220 mg, 9.2 mmol, 2.0 eq). React at room temperature for 16 hours. TLC detection showed the starting material had disappeared. Concentrate the mixture, add water (15 mL) to the residue, adjust the pH to 6-7 with 4N hydrochloric acid, extract the aqueous phase with DCM (20 mL × 3), combine the organic phases, wash once with saturated brine, dry over anhydrous sodium sulfate, concentrate, and obtain the crude product. Column chromatography purification of the crude product yielded 610 mg of the target analyte (yield: 54.1%). ESI-MS (+): m / z = 246.19 [M+1].

[0165] Example 9: Synthesis of compound DSC207-07

[0166] Reaction formula:

[0167] Preparation method:

[0168] DSC207-05-1 (1 g, 3.77 mmol, 1 eq) was added to methanol (10 mL), and the system was cooled to -10 °C. Lithium aluminum hydride-4d (174.1 mg, 4.15 mmol, 1.1 eq) was added, and the mixture was allowed to react at room temperature for 4 hours. Thin-layer chromatography showed complete reaction. The system was poured into an aqueous solution (50 mL), and the aqueous phase was extracted with DCM (50 mL × 3). The organic phases were combined, dried over sodium sulfate, concentrated, and the residue was purified by column chromatography to give 462.7 mg of the target analyte (yield: 51.3%). ESI-MS(+): m / z = 240.21 [M+1]

[0169] Example 10: Synthesis of compound DSC207-08

[0170] Reaction formula:

[0171] Preparation method:

[0172] Step 1: Preparation of compound DSC207-08-1

[0173] ZJT-3 (1.5 g, 14.5 mmol, 1.45 eq) was added to DMSO (30 mL), the system was cooled to -10 °C, and then potassium carbonate (2.76 g, 20.0 mmol, 2 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 10 minutes, and then DSC207-03-3 (2.0 g, 10.0 mmol, 1 eq) was added. After the addition was complete, the mixture was reacted at room temperature for 3 hours.

[0174] Thin-layer chromatography showed the reaction was complete. The system was poured into water, the pH was adjusted to 3-4 with 1N hydrochloric acid, and extracted with ethyl acetate (25ml × 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by column chromatography to give 1.3g of the target analyte (yield: 48.6%). ESI-MS(+): m / z = 268.26 [M+1].

[0175] Step 2: Preparation of compound DSC207-08

[0176] DSC207-08-1 (1.0 g, 3.74 mmol, 1 eq) was added to methanol (10 mL), and the system was cooled to -10 °C. Sodium borohydride (566.0 mg, 15.0 mmol, 4 eq) was added, and the mixture was allowed to react at room temperature for 4 hours. Thin-layer chromatography showed that the reaction was complete. The system was poured into an aqueous solution (50 mL), and the aqueous phase was extracted with DCM (50 mL × 3). The organic phases were combined, dried over sodium sulfate, concentrated, and the residue was purified by column chromatography to give 418 mg of the target analyte (yield: 46.7%). ESI-MS (+): m / z = 240.36 [M+1].

[0177] Example 11: Synthesis of compound DSC207-09

[0178] Reaction formula:

[0179] Preparation method:

[0180] Step 1: Preparation of compound ZJT-4

[0181] Under nitrogen atmosphere, 2.8 g (28.6 mmol, 1 eq) of 3-methyl-2-pyrazolin-5-one was added to 30 mL of heavy water, followed by potassium carbonate (4.0 g, 28.6 mmol, 1 eq). The mixture was stirred at room temperature for 15 min. Liquid chromatography-mass spectrometry (LC-MS) analysis showed the reaction was complete. The reaction mixture was extracted with DCM (50 mL × 3), and the organic phases were combined, dried over anhydrous magnesium sulfate, filtered, and concentrated to give 2.1 g of the target compound (yield: 73.3%).

[0182] ESI-MS(+): m / z = 101.11 [M+1]

[0183] Step 2: Preparation of compound DSC207-09-1

[0184] ZJT-4 (1.45 g, 14.5 mmol, 1.45 eq) was added to DMSO (30 mL), and the system was cooled to -10 °C. Potassium carbonate (2.76 g, 20.0 mmol, 2 eq) was then added. After the addition was complete, the mixture was stirred at room temperature for 10 minutes. DSC207-03-3 (2.0 g, 10.0 mmol, 1 eq) was then added, and the mixture was allowed to react at room temperature for 3 hours. Thin-layer chromatography showed that the reaction was complete. The system was poured into water, and the pH was adjusted to 3-4 with 1 N hydrochloric acid. Extraction was performed with ethyl acetate (20 mL × 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain 1.2 g of the target analyte (yield: 45.4%).

[0185] ESI-MS(+): m / z=265.34[M+1].

[0186] Step 3: Preparation of compound DSC207-09

[0187] DSC207-09-1 (988 mg, 3.74 mmol, 1 eq) was added to methanol (10 mL), the system was cooled to -10 °C, sodium borohydride (566 mg, 15 mmol, 4 eq) was added, and after the addition was complete, the mixture was brought to room temperature and reacted for 4 hours.

[0188] Thin-layer chromatography showed that the reaction was complete. The system was poured into an aqueous solution (50 mL), and the aqueous phase was extracted with DCM (50 mL × 3). The organic phases were combined, dried over sodium sulfate, and concentrated to give 473 mg of a colorless oil (yield: 53.5%). ESI-MS(+): m / z = 237.41 [M+1].

[0189] Example 12: Synthesis of compound DSC207-10

[0190] Reaction formula:

[0191] Preparation method:

[0192] Step 1: Preparation of compound DSC207-10-3

[0193] ZJT-3 (6.0 g, 58.2 mmol, 1.45 eq) was added to DMSO (120 mL), the system was cooled to -10 °C, and then potassium carbonate (11.1 g, 80.0 mmol, 2.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 10 minutes, and then DSC207-01-2 (6.6 g, 40.0 mmol, 1 eq) was added. After the addition was complete, the mixture was reacted at room temperature for 3 hours.

[0194] Thin-layer chromatography showed that the reaction was complete. The system was poured into water, the pH was adjusted to 3-4 with 1N hydrochloric acid, and extracted with ethyl acetate (100mL×3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to give 6.1g of the target substance (yield: 65.6%).

[0195] ESI-MS(+): m / z=233.22[M+1].

[0196] Step 2: Preparation of compound DSC207-10-2

[0197] Add DSC207-10-3 (4.0 g, 17.4 mmol, 1 eq) to DCM (40 ml), cool the system to -10 °C, add m-CPBA (6.0 g, 34.8 mmol, 2 eq) in portions, and react for another 30 min after the addition is complete.

[0198] Thin-layer chromatography showed that the reaction was complete. The system was heated to 25°C and poured into an aqueous sodium sulfite solution (50 mL). The mixture was separated, and the aqueous phase was extracted with DCM (50 mL × 3). The organic phases were combined, dried over sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by column chromatography to obtain 3.02 g of the target analyte (yield: 69.9%).

[0199] ESI-MS(+): m / z=249.11[M+1].

[0200] Step 3: Preparation of compound DSC207-10-1

[0201] DSC207-10-2 (3.0 g, 12.1 mmol, 1 eq) was added to acetic anhydride (30 mL), and the system was heated to 130 °C and reacted for 5 hours. Thin-layer chromatography showed that the reaction was complete. The system was cooled to room temperature and concentrated to obtain the crude product. The crude product was purified by column chromatography to obtain 2.1 g of the target analyte (yield: 60.1%).

[0202] ESI-MS(+): m / z=290.39[M+1].

[0203] Step 4: Preparation of compound DSC207-10

[0204] Add DSC207-10-1 (1.33 g, 4.6 mmol, 1 eq) to a single-necked flask, add methanol (15 mL), and add lithium hydroxide (220 mg, 9.2 mmol, 2.0 eq). React at room temperature for 16 hours. The starting material disappeared as detected by TLC. Concentrate the sample, add water (15 mL) to the residue, adjust the pH to 6-7 with 4N hydrochloric acid, extract the aqueous phase with DCM (20 mL × 3), combine the organic phases, wash once with saturated brine, dry over anhydrous sodium sulfate, concentrate, and obtain the crude product. Column chromatography (dichloromethane:methanol = 30:1) of the crude product yielded 481 mg of the target analyte (yield: 42.3%). ESI-MS (+): m / z = 248.72 [M+1].

[0205] Example 13: Synthesis of compound DSC207-11

[0206] Reaction formula:

[0207] Preparation method:

[0208] Step 1: Preparation of compound DSC207-11-1

[0209] DSC207-11-SM (4.12 g, 38.16 mmol, 1 eq) was added to DCM (50 mL), and the system was cooled to -10 °C. m-CPBA (7.24 g, 41.97 mmol, 1.1 eq) was added in portions, and the reaction was allowed to proceed for another 40 minutes after the addition was complete. Thin-layer chromatography showed that the reaction was complete. The system was heated to 25 °C and poured into an aqueous sodium sulfite solution (50 mL). The mixture was separated, and the aqueous phase was extracted with DCM (50 mL × 3). The organic phases were combined, dried over sodium sulfate, and concentrated to give 4.3 g of the target compound (yield: 90.8%). ESI-MS(+): m / z = 125.19 [M+1].

[0210] Step 2: Preparation of compound DSC207-11-2

[0211] DSC207-11-1 (4.22 g, 34.0 mmol, 1 eq) was added to toluene (50 mL), and the system was cooled to -10 °C. Phosphorus oxychloride (6.25 g, 40.8 mmol, 1.2 eq) was then added. After the addition was complete, the system was heated to 90 °C and reacted for 16 hours. Thin-layer chromatography showed that the reaction was complete. The mixture was concentrated, and the residue was dissolved in DCM (20 mL). The pH was adjusted to 8-9 with sodium bicarbonate aqueous solution. The mixture was separated, and the aqueous phase was extracted with DCM (50 mL × 3). The organic phases were combined, washed once with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was subjected to column chromatography (petroleum ether: ethyl acetate = 20:1) to obtain 2.2 g of the target analyte (yield: 45.4%). ESI-MS (+): m / z = 143.13 [M+1].

[0212] Step 3: Preparation of compound DSC207-11-3

[0213] ZJT-2 (1.47 g, 14.5 mmol, 1.45 eq) was added to DMSO (30 mL), and the system was cooled to -10 °C. Potassium carbonate (2.75 g, 20 mmol, 2 eq) was then added. After the addition was complete, the mixture was stirred at room temperature for ten minutes. DSC207-11-2 (1.43 g, 10.0 mmol, 1 eq) was then added, and the mixture was allowed to react at room temperature for 3 hours. Thin-layer chromatography showed that the reaction was complete. The system was poured into water, and the pH was adjusted to 3-4 with 1 N hydrochloric acid. Extraction was performed with ethyl acetate (20 mL × 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by column chromatography to obtain 1.63 g of the target analyte (yield: 78.7%). ESI-MS (+): m / z = 208.21 [M+1].

[0214] Step 4: Preparation of compound DSC207-11-4

[0215] DSC207-11-3 (1.6 g, 7.72 mmol, 1 eq) was added to DCM (40 mL), and the system was cooled to -10 °C. m-CPBA (2.66 g, 15.44 mmol, 2 eq) was added in portions, and the reaction was allowed to proceed for another 30 minutes after the addition was complete. Thin-layer chromatography showed that the reaction was complete. The system was heated to 25 °C and poured into an aqueous sodium sulfite solution (50 mL). The mixture was separated, and the aqueous phase was extracted with DCM (50 mL × 3). The organic phases were combined, dried over sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by column chromatography to obtain 1.02 g of the target analyte (yield: 59.2%). ESI-MS (+): m / z = 224.31 [M+1].

[0216] Step 5: Preparation of compound DSC207-11-5

[0217] DSC207-11-4 (1.0 g, 4.18 mmol, 1 eq) was added to toluene (25 mL), and the system was cooled to -10 °C. Phosphorus oxychloride (0.77 g, 5.02 mmol, 2 eq) was then added. After the addition was complete, the system was heated to 90 °C and reacted for 16 hours. Thin-layer chromatography showed that the reaction was complete. The mixture was concentrated, and the residue was dissolved in DCM (20 mL). The pH was adjusted to 8-9 with sodium bicarbonate aqueous solution. The mixture was separated, and the aqueous phase was extracted with DCM (30 mL × 3). The organic phases were combined, washed once with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by column chromatography to obtain 0.52 g of the target analyte (yield: 51.5%). ESI-MS (+): m / z = 242.23 [M+1].

[0218] Step 6: Preparation of compound DSC207-11

[0219] DSC207-11-5 (0.5 g, 2.07 mmol, 1 eq) and (2.41 g, 10 mmol, 1 eq) were added to ethanol (25 mL). The system was cooled to -10 °C, and sodium ethoxide (0.16 g, 2.35 mmol, 1.1 eq) and (748.5 mg, 11 mmol, 1.1 eq) were added. The reaction was allowed to proceed for another 50 minutes. Thin-layer chromatography showed that the reaction was complete. The system was poured into 1 N hydrochloric acid aqueous solution (35 mL), and the aqueous phase was extracted with ethyl acetate (50 mL × 3). The organic phases were combined, dried over sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by column chromatography to obtain 0.22 g of the target analyte (yield 42.3%). ESI-MS (+): m / z = 252.26 [M+1].

[0220] Example 14: Synthesis of compound DSC207-16

[0221] Reaction formula:

[0222] Step 1: Preparation of compound DSC207-16-1

[0223] The compound DSC207-16-SM (16.0 g, 105.2 mmol) was dissolved in ethanol (250 mL). The system was cooled to 0 °C, and 2 drops of sulfuric acid were added. After the addition was complete, the system was allowed to react at room temperature for 3 hours. Thin-layer chromatography showed that the reaction was essentially complete. The system was concentrated, and the residue was poured into water and extracted with ethyl acetate (150 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give 14.4 g of the target compound (yield: 76.5%).

[0224] ESI-MS(+): m / z = 181.78 [M+1]

[0225] Step 2: Preparation of compound DSC207-16-2

[0226] DSC207-16-1 (7.58 g, 42.12 mmol, 1 eq) was added to dichloromethane (150 mL), and the system was cooled to -10 °C. m-CPBA (8.72 g, 50.54 mmol, 1.2 eq) was added in portions, and the reaction was allowed to proceed at room temperature for 8 hours after the addition was complete. Thin-layer chromatography showed that the reaction was complete. The system was heated to 25 °C and poured into an aqueous sodium sulfite solution (50 mL). The mixture was separated, and the aqueous phase was extracted with dichloromethane (80 mL × 3). The organic phases were combined, dried over sodium sulfate, and concentrated to give 7.0 g of the target compound (yield: 84.7%).

[0227] ESI-MS(+): m / z=197.41[M+1].

[0228] Step 3: Preparation of compound DSC207-16-3

[0229] DSC207-16-2 (5.38 g, 27.44 mmol, 1 eq) was added to toluene (50 mL), and the system was cooled to -10 °C. Phosphorus oxychloride (5.1 g, 32.93 mmol, 1.2 eq) was then added. After the addition was complete, the system was heated to 90 °C and reacted for 16 hours. Thin-layer chromatography showed that the reaction was complete. The mixture was concentrated, and the residue was dissolved in DCM (25 mL). The pH was adjusted to 8-9 with sodium bicarbonate aqueous solution. The mixture was separated, and the aqueous phase was extracted with DCM (50 mL × 3). The organic phases were combined, washed once with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give 4.8 g of the target compound (yield 81.5%). ESI-MS (+): m / z = 215.17 [M+1].

[0230] Step 4: Preparation of compound DSC207-16

[0231] ZJT-2 (1.47 g, 14.5 mmol, 1.45 eq) was added to DMSO (50 mL), and the system was cooled to -10 °C. Potassium carbonate (2.75 g, 20.0 mmol, 2 eq) was then added. After the addition was complete, the mixture was stirred at room temperature for ten minutes. DSC207-16-3 (2.15 g, 10.0 mmol, 1 eq) was then added, and the mixture was allowed to react at room temperature for 3 hours. Thin-layer chromatography showed that the reaction was complete. The system was poured into water, and the pH was adjusted to 3-4 with 1 N hydrochloric acid. Extraction was performed with ethyl acetate (50 mL × 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the residue purified by column chromatography to obtain 1.3 g of the target compound (yield: 46.5%).

[0232] ESI-MS(+): m / z=280.33[M+1].

[0233] Example 15: Synthesis of compound DSC207-20

[0234] Reaction formula:

[0235] Preparation method:

[0236] Step 1: Preparation of compound DSC207-20-1

[0237] ZJT-3 (2.32 g, 22.5 mmol, 1.5 eq) was added to DMSO (100 mL), and the system was cooled to -10 °C. Potassium carbonate (4.1 g, 30 mmol, 2 eq) was then added. After the addition was complete, the mixture was stirred at room temperature for ten minutes. DSC207-03-3 (3.0 g, 15 mmol, 1 eq) was then added, and the mixture was reacted at room temperature for 3 hours. Thin-layer chromatography showed that the reaction was complete. The system was poured into water, and the pH was adjusted to 3-4 with 1 N hydrochloric acid. Extraction was performed with ethyl acetate (80 mL × 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Column purification yielded 1.79 g of the target compound (yield: 44.7%). ESI-MS (+): m / z = 268.29 [M+1]

[0238] Step 2: Preparation of compound DSC207-20

[0239] DSC207-20-1 (1.0 g, 3.74 mmol, 1 eq) was added to tetrahydrofuran (10 mL), and the system was cooled to -10 °C. Lithium aluminum hydride-4d (172 mg, 4.11 mmol, 1.1 eq) was added, and the mixture was allowed to react at room temperature for 4 hours. Thin-layer chromatography showed that the reaction was complete. The system was poured into an aqueous solution (50 mL), and the aqueous phase was extracted with DCM (40 mL × 3). The organic phases were combined, dried over sodium sulfate, concentrated, and the residue was purified by column chromatography to give 359.2 mg of the target compound (yield: 39.8%). ESI-MS(+): m / z = 242.36 [M+1].

[0240] Example 16: Synthesis of compound DSC207-21

[0241] Reaction formula:

[0242] Preparation method:

[0243] Step 1: Preparation of compound DSC207-21-1

[0244] DSC207-03-4 (1.0 g, 3.81 mmol, 1 eq) was added to DCM (10 mL), and the system was cooled to -10 °C. Diisobutylaluminum hydride (DIBAL-H, 1 mol / L in DCM) (4.2 mL, 4.2 mmol, 1.1 eq) was added, and the mixture was allowed to react at room temperature for 4 hours. Thin-layer chromatography showed that the reaction was complete. The system was poured into an aqueous solution (50 mL), and the aqueous phase was extracted with DCM (40 mL × 3). The organic phases were combined, dried over sodium sulfate, concentrated, and the residue was purified by column chromatography to give 521 mg of the target analyte (yield: 58.9%).

[0245] ESI-MS(+): m / z=233.45[M+1].

[0246] Step 2: Preparation of compound DSC207-21

[0247] DSC207-21-1 (0.5 g, 2.15 mmol, 1 eq) was added to ethanol (10 mL), followed by DSC207-21-SM (316 mg, 3.22 mmol, 1.5 eq). The mixture was heated to 80 °C and reacted for 4 hours. Thin-layer chromatography showed that the reaction was complete. The solution was concentrated to obtain the crude product. Column purification yielded 250.5 mg of the target compound (yield: 37.3%). ESI-MS (+): m / z = 313.27 [M+1].

[0248] Example 17: Synthesis of compound DSC207-22

[0249] Reaction formula:

[0250] Preparation method:

[0251] Step 1: Preparation of compound DSC207-22-1

[0252] DSC207-16 (1.0 g, 3.58 mmol, 1 eq) was added to DCM (30 mL), and the system was cooled to -10 °C. DIBAL-H (1 mol / L in DCM) (4.14 mL, 4.14 mmol, 1.1 eq) was added, and the mixture was allowed to react at room temperature for 4 hours. Thin-layer chromatography showed that the reaction was complete. The system was poured into an aqueous solution (50 mL), and the aqueous phase was extracted with DCM (30 mL × 3). The organic phases were combined, dried over sodium sulfate, concentrated, and the residue was purified by column chromatography to give 526 mg of the target analyte (yield: 62.4%). ESI-MS (+): m / z = 236.36 [M+1].

[0253] Step 2: Preparation of compound DSC207-22

[0254] DSC207-22-1 (505 mg, 2.15 mmol, 1 eq) was added to ethanol (20 mL), followed by DSC207-21-SM (0.319 mg, 3.25 mmol, 1.5 eq). The mixture was heated to 80 °C and reacted for 4 hours. Thin-layer chromatography showed that the reaction was complete. The mixture was concentrated, and the residue was purified by column chromatography to obtain 215 mg of the target analyte (yield: 31.7%). ESI-MS(+): m / z = 316.37 [M+1].

[0255] Example 18: Synthesis of compound DSC207-02

[0256] Reaction formula:

[0257] Preparation method:

[0258] Step 1: Preparation of compound DSC207-02-1

[0259] Under nitrogen atmosphere, 12.36 g (114.4 mmol) of 2,5-dimethylpyrazine was added to 120 mL of heavy water, followed by 20 mL of 40% deuterated sodium hydroxide solution, and refluxed for five days. Liquid chromatography-mass spectrometry (LC-MS) analysis showed the reaction was complete.

[0260] The reaction mixture was extracted with DCM (150 mL × 3), the organic phases were combined, dried over anhydrous magnesium sulfate, filtered, and concentrated to give 9.04 g of a pale yellow oily liquid (yield: 69.3%).

[0261] ESI-MS(+): m / z = 115.17 [M+1]

[0262] Step 2: Preparation of compound DSC207-02-2

[0263] Add DSC207-02-1 (8.7 g, 76.32 mmol, 1 eq) to DCM (50 ml), cool the system to -10 °C, add m-CPBA (14.48 g, 83.94 mmol, 1.1 eq) in portions, and react for another 40 min after the addition is complete.

[0264] Thin-layer chromatography showed that the reaction was complete. The system was heated to 25°C and poured into an aqueous sodium sulfite solution (50 mL). The mixture was separated, and the aqueous phase was extracted with DCM (50 mL × 3). The organic phases were combined, dried over sodium sulfate, and concentrated to give a yellow oily substance (8.4 g, 84.6%).

[0265] ESI-MS(+): m / z=131.18[M+1].

[0266] Step 3: Preparation of compound DSC207-02-3

[0267] DSC207-02-2 (4.42 g, 34 mmol, 1 eq) was added to toluene (25 ml), the system was cooled to -10 °C, and then phosphorus oxychloride (6.25 g, 40.8 mmol, 1.2 eq) was added. After the addition was complete, the system was heated to 90 °C and reacted for 16 hours.

[0268] Thin-layer chromatography showed that the reaction was complete. The mixture was concentrated, and the residue was dissolved in DCM (20 ml). The pH was adjusted to 8-9 with sodium bicarbonate aqueous solution. The mixture was separated, and the aqueous phase was extracted with DCM (50 ml * 3). The organic phases were combined, washed once with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product.

[0269] The crude product was subjected to column chromatography (petroleum ether: ethyl acetate = 20:1) to give 3.7 g of a yellow solid (yield: 73.2%).

[0270] ESI-MS(+): m / z=149.29[M+1].

[0271] Step 4: Preparation of compound DSC207-02-4

[0272] ZJT-2 (2.92 g, 29 mmol, 1.45 eq) was added to DMSO (30 ml), the system was cooled to -10 °C, and then potassium carbonate (5.5 g, 39.8 mmol, 2 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 10 minutes, and then DSC207-02-3 (2.96 g, 19.92 mmol, 1 eq) was added. After the addition was complete, the mixture was reacted at room temperature for 3 hours.

[0273] Thin-layer chromatography showed that the reaction was complete. The system was poured into water, the pH was adjusted to 3-4 with 1N hydrochloric acid, and extracted with ethyl acetate (50ml*3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product.

[0274] The crude product was subjected to column chromatography (petroleum ether: ethyl acetate = 10:1) to give 3.93 g of white solid (yield: 92.7%).

[0275] ESI-MS(+): m / z=214.29[M+1].

[0276] Step 5: Preparation of compound DSC207-02-5

[0277] Add DSC207-02-4 (3.7 g, 17.36 mmol, 1 eq) to DCM (40 ml), cool the system to -10 °C, add m-CPBA (5.13 g, 34.73 mmol, 2 eq) in portions, and react for another 30 min after the addition is complete.

[0278] Thin-layer chromatography showed that the reaction was complete. The system was heated to 25°C and poured into an aqueous sodium sulfite solution (50 mL). The mixture was separated, and the aqueous phase was extracted with DCM (50 mL × 3). The organic phases were combined, dried over sodium sulfate, and concentrated to obtain the crude product.

[0279] The crude sample was subjected to column chromatography (petroleum ether: ethyl acetate = 10:1) to give a yellow solid (3.1 g, 77.8%).

[0280] ESI-MS(+): m / z=230.39[M+1].

[0281] Step 6: Preparation of compound DSC207-02

[0282] DSC207-02-5 (3 g, 13.08 mmol, 1 eq) was added to acetic anhydride (30 mL), and the system was heated to 130 °C and reacted for 5 hours. Thin-layer chromatography showed that the reaction was complete. The system was cooled to room temperature and concentrated to obtain the residue. The residue was added to 30 mL of an aqueous solution of 2N hydrochloric acid, and stirred at room temperature for 2 hours. Thin-layer chromatography showed that the reaction was complete. The aqueous phase was extracted with DCM (50 mL × 3), the organic phases were combined, dried over sodium sulfate, and concentrated to obtain the crude product. The crude product was subjected to column chromatography (petroleum ether: ethyl acetate = 20:1) to give a yellow solid (3.1 g, 77.8%).

[0283] ESI-MS(+): m / z = 241.3 [M+1].

[0284] The compounds of the following examples were synthesized using the same method as in the above embodiments, either commercially available compounds or intermediate compounds appropriately synthesized from commercially available compounds.

[0285] Example 19: Protective effect against L-glutamate-induced PC12 cell damage

[0286] X1901 structure:

[0287] Cells: Rat adrenal pheochromocytoma cells PC-12 (highly differentiated), also known as PC-12Adh (National Model and Characteristic Experimental Cell Resource Bank / Chinese Academy of Sciences Cell Bank Stem Cell Bank, catalog number SCSP-5259)

[0288] Environment: The cell culture incubator was maintained at 37°C and 5% CO2. Culture medium (RPMI 1640, Gibco): Fetal bovine serum (FBS) (10091-148, Gibco) (v:v) = 9:1.

[0289] Main instruments and equipment:

[0290] Main reagents:

[0291] Preparation of modeling agent solution (L-glutamic acid): Weigh an appropriate amount of L-glutamic acid powder, dissolve it in RPMI 1640 complete culture medium (containing 10% FBS) to a concentration of 10 mmol / L, sonicate for 15 min, and then sonicate in a 37℃ water bath for 15 min until completely dissolved. Prepare fresh before use.

[0292] Preparation of test solution series: Dissolve an appropriate amount of sample in DMSO to 100 mmol / L as a stock solution and store at -20℃ for later use. Immediately before use, dilute the above-prepared modeling agent solution (final L-glutamic acid concentration 10 mmol / L) 1000 times with RPMI 1640 whole medium (containing 10% FBS) to 100 μmol / L. After thorough mixing, take an appropriate amount and dilute it 1 / 2 with the above-prepared modeling agent solution to 50 μmol / L. Then, continue to dilute 1 / 2 to obtain test solution series concentrations (25, 12.5, 6.25, 3.125, 1.6, 0.8, 0.4, 0.2 μmol / L).

[0293] Test method:

[0294] PC12 cells in the logarithmic growth phase were harvested from in vitro cultured cells (96-well cell culture plate, 5 × 10⁶ cells / well). 3 The experiment was conducted using cells / well, 100 μL / well, 24 h after seeding. The experiment included a blank control group, a model group, a control compound group (X1901), and a test substance group. After removing the original culture medium, the control group was given 100 μL / well of RPMI 1640 complete medium (containing 10% FBS), the model group was given 100 μL / well of the above-prepared modeling agent solution, and the test substance group was given 100 μL / well of a series of concentrations of the test substance (0.2, 0.4, 0.8, 1.6, 3.125, 6.25, 12.5, 25, 50, 100 μmmol / L, a total of 10 concentration groups). Each group had 5 replicates. After adding all solutions, the cells were immediately incubated at 37℃ in a 5% CO2 incubator. After 24 h of culture, the viability of PC12 cells was detected using the CCK-8 assay. Calculate the cell death inhibition rate at different concentrations of the test substance. Cell death inhibition rate = (OD 给药组 -OD 模型组 ) / (OD 空白对照组 -OD 模型组 ×100%, where OD represents the absorbance at 450 nm measured using a microplate reader for the corresponding group. The EC50 of the corresponding test substance was calculated using GraphPad Prism 6.0 software. 50 (50% effective concentration).

[0295] The experimental results are shown in Table 1.

[0296] Table 1. Protective effect of the compounds of this invention against L-glutamate-induced PC12 cell damage.

[0297] Data show that, compared with the comparative compounds, the compounds of the present invention have very strong protective activity against L-glutamate-induced PC12 cell damage, especially DSC207-L01, DSC207-L02, DSC207-L03, DSC207-L04 and DSC207-L05, among which compound DSC207-L05 has the strongest protective activity, which is about 7.5 times that of the comparative compound X1901.

[0298] Example 20: Pharmacological efficacy test in a rat model of cerebral ischemia-reperfusion.

[0299] Animals: SD rats, SPF grade, male, sourced from Shanghai Silex Laboratory Animal Co., Ltd.

[0300] Number of animals included: 72; animal weight at the start of the experiment: approximately 180-200g.

[0301] Test method:

[0302] Grouping: Rats were acclimatized for 7 days. After 4 days of acclimatization, i.e., 3 days before surgery, all rats in each group began behavioral training (balance beam, rotisserie). SD rats were evenly divided into 9 groups according to body weight: sham-operated group, model group, positive control drug (Edaravone Dexborneol Injection) group, control group (X1901 group), DSC207-01 group, DSC207-02 group, DSC207-03 group, DSC207-04 group, and DSC207-05 group, with a total of 8 rats in each group.

[0303] Model Establishment: Rats were fasted for 12-14 hours before surgery, but had free access to water. Before surgery, all groups of animals were anesthetized with isoflurane, preserving spontaneous respiration. They were fixed in a supine position on a rat board, with hair removed from the midline of the neck and disinfected with 75% alcohol. A surgical incision was made on the ventral side of the neck midline, separating the muscle and fascia along the inner edge of the sternocleidomastoid muscle. The right common carotid artery, external carotid artery, and internal carotid artery were then separated. A small oblique incision was made on the external carotid artery approximately 0.5 cm proximal to the ligation site using vascular scissors. The proximal end of the external carotid artery was pulled until it was aligned with the internal carotid artery. The suture occluded the rat and slowly advanced 1.8 cm towards the internal carotid artery through the incision on the right external carotid artery, marking the bifurcation of the common carotid artery. When slight resistance was felt during advancement, the middle cerebral artery was blocked. The suture occluded 2 hours after infarction, completing the cerebral ischemia-reperfusion injury model. The sham-operated group only underwent vascular dissection. Twenty minutes after infarction, cerebral blood flow was assessed using Doppler flowmetry. A model was considered successful and enrolled in the experiment if the difference in blood flow between the left and right hemispheres (ROI%) was greater than 48%. Incandescent lamps were used to maintain rectal temperature during the procedure.

[0304] Drug administration: The drugs for each group were administered intravenously 0.5 hours after cerebral infarction in the animals. The sham-operated group and the model group were given the same volume of blank solvent (8% propylene glycol + 92% physiological saline). Grouping and detailed drug administration information are shown in Table 2.

[0305] Table 2. Trial grouping and drug administration information

[0306] Twenty-four hours after reperfusion, each animal underwent Zea-longa neurological function assessment. At the end of the experiment (24 hours after reperfusion), changes in cerebral blood flow were measured in all animals using Doppler flowmeter to evaluate the ameliorative effect of the test substance on ischemic stroke. Behavioral function tests (balance beam) were performed. One hour later, all animals were euthanized, and serial coronal sections of the whole brain were prepared for TTC staining and photography. The infarct area was measured using Image-J software, the percentage of the infarct area to the total brain area was calculated, and the infarct improvement rate was calculated.

[0307] Zea-Longa scoring criteria: No neurological deficits: 0 points; Inability to fully extend the forepaw on the paralyzed side: 1 point; Turning in circles towards the paralyzed side while walking: 2 points; Leaning towards the paralyzed side while walking: 3 points; Inability to walk automatically, with signs of loss of consciousness: 4 points.

[0308] Infarct area ratio = Total infarct area ÷ Total area of ​​whole brain slices × 100%;

[0309] Infarction improvement rate = (Infarction area ratio in the model group - Infarction area ratio in the treatment group) ÷ Infarction area ratio in the model group × 100%

[0310] Experimental data are expressed as Mean ± SD. SPSS 21.0 software was used for statistical analysis. A p < 0.05 was considered statistically significant.

[0311] The efficacy test results for rats with ischemic stroke are shown in Table 3.

[0312] Table 3. Results of the pharmacodynamic test in rats with ischemic stroke. ## p<0.01 vs. sham surgery group; *p<0.05, **p<0.01 vs. model group

[0313] The above data indicate that, in this efficacy experiment of a single intravenous injection of the test substance in a rat model of MCAO, the test substance groups (DSC207-01, DSC207-02, DSC207-03, DSC207-04, and DSC207-05) all effectively improved the Zea-Longa score and balance beam time of MCAO rats compared with the positive drug group (Xenbisin) and the control group (X1901). They also significantly improved the difference in blood flow between the left and right sides of the brain in MCAO rats, significantly reduced the cerebral infarction area, and increased the cerebral infarction improvement rate in rats. Among them, DSC207-05 showed the best performance.

[0314] Example 21: Distribution of drug in brain and spinal cord tissues in rats via gavage

[0315] Animals: Male SD rats, SPF grade, sourced from Beijing Vital River Co., Ltd.

[0316] The number of animals included in the group was 36, and their weight was approximately 200±20g at the start of the experiment.

[0317] Test method:

[0318] Male SD rats were randomly divided into four groups according to body weight: a control group (X1901 group), a DSC207-02 group, a DSC207-03 group, and a DSC207-05 group, with nine rats in each group. Animals were fasted for 12-14 hours before administration, but water was allowed. On the day of the experiment, the animals were administered the drug by gavage at a dose of 13.40 mg / kg (5 mL / kg, 5% DMSO + 95% 0.5% MC solution). Three animals from each group were euthanized at 10 min, 60 min, and 3 h after the single oral gavage administration. Brain and spinal cord tissue samples were collected (care was taken to remove blood with filter paper and remove blood vessels as much as possible) and stored at -80℃ for later analysis.

[0319] Throughout the experiment, the experimental animals were observed in their general condition. This included: changes in the rats' food and water intake, weight changes, any abnormalities in coat color, behavior and mental state, any abnormal secretions from the eyes, ears, mouth, and nose, and any abnormalities in urination and defecation. Any abnormalities were immediately recorded, and the causes were analyzed.

[0320] Detection method: Plasma protein precipitation-LC / MS / MS method was used to detect the original drug content in brain tissue and spinal cord.

[0321] The results of the brain and spinal cord tissue distribution test after oral administration to rats are shown in Table 4.

[0322] Table 4. Average concentrations (ng / g) of the parent drug in brain and spinal cord tissues after a single gavage administration.

[0323] Data show that after oral administration to rats, both the compound of this invention and X1901 rapidly distributed in brain and spinal cord tissues. Compared to X1901, the compound of this invention showed significantly higher distribution levels in brain and spinal cord tissues. Even 3 hours after administration, the compound of this invention maintained high exposure levels in brain and spinal cord tissues, significantly higher than that of X1901. In conclusion, the compound of this invention exhibits higher exposure levels in brain and spinal cord tissues and a longer retention time in these tissues, with DSC207-05 showing the best results.

[0324] Example 22: Rat Pharmacokinetic Study

[0325] Grouping and Administration: Eighteen male SD rats were divided into 6 groups of 3 rats each. Animals were fasted for 12-14 hours before administration, but water was allowed. The control group (X1901 group), DSC207-01 group, DSC207-02 group, DSC207-03 group, DSC207-04 group, and DSC207-05 group were all administered the drug via tail vein injection. On the day of the experiment, the drugs were administered sequentially according to the animals' body weight at a dose of 5 mg / kg (administration volume of 5 mL / kg).

[0326] Plasma collection: Within 0.5 h before a single intravenous injection (0 h), and at 0.083 h, 0.25 h, 0.50 h, 1.0 h, 3.0 h, 6.0 h, 9.0 h, and 24.0 h after administration, 200 μL of blood was collected from the orbital vein of rats and placed in EDTA-K2 anticoagulant tubes. The tubes were then placed in an ice bath and centrifuged at 4000 rpm for 10 min. The plasma was transferred to 1.5 mL centrifuge tubes and stored at -80℃ for analysis. Animals were fed 4 h after administration, but water was not restricted throughout the process.

[0327] Data calculation: Drug concentration was analyzed using LC-MS / MS, and pharmacokinetic parameters were calculated using DAS3.2.7 software.

[0328] Pharmacokinetic parameters of rats administered via intravenous injection are shown in Table 5.

[0329] Table 5. Average pharmacokinetic parameters of rats administered via intravenous injection.

[0330] Data shows that, unexpectedly, the half-life of the compounds of this invention is significantly shorter than that of X1901 under similar in vivo exposure and clearance conditions. Common sense dictates that the substitution of hydrogen atoms with deuterium atoms in drug structures often significantly prolongs the drug's half-life. However, the compounds of this invention exhibit a shortened half-life, a characteristic that is highly advantageous in the treatment of cardiovascular and cerebrovascular diseases, as it can reduce drug accumulation in the body, thereby significantly reducing or avoiding the damaging side effects of such drugs on liver and kidney function.

[0331] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications and improvements without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention shall be defined by the claims.

Claims

1. A novel deuterium-containing pyrazolinone compound, tautomer, stereoisomer, prodrug, and pharmaceutically acceptable salt thereof as shown in formula (I) and / or formula (II): In formula (I) and / or formula (II), R1 is selected from hydrogen, deuterium, C1-C8 alkyl groups substituted or unsubstituted with one or more substituents, phenyl groups substituted or unsubstituted with one or more substituents, or pyridyl groups substituted or unsubstituted with one or more substituents, wherein, The above substituents are selected from deuterium or halogens; R2 is selected from hydrogen, deuterium, C1-C8 alkyl group substituted or unsubstituted with one or more substituents, phenyl group substituted or unsubstituted with one or more substituents, or pyridyl group substituted or unsubstituted with one or more substituents, wherein the substituents are selected from deuterium or halogens; R3 is selected from hydrogen or deuterium; R4 is selected from cyclohexyl groups substituted or unsubstituted with one or more substituents, benzyl groups substituted or unsubstituted with one or more substituents, naphthyl groups substituted or unsubstituted with one or more substituents, heterocyclic groups substituted or unsubstituted with one or more substituents, or... The substituents mentioned above are selected from deuterium or halogens; R X1 Selected from halogens, or C1-C8 alkyl groups substituted or unsubstituted by one or more substituents, wherein the substituents are selected from deuterium or halogens; R X2 Selected from halogens, or C1-C8 alkyl groups substituted or unsubstituted by one or more substituents, wherein the substituents are selected from deuterium or halogens; R X3 Selected from halogens, C1-C8 alkyl groups substituted or unsubstituted with one or more substituents, and C1-C8 alkoxy groups substituted or unsubstituted with one or more substituents. The substituents mentioned above are selected from deuterium and hydroxyl groups; The above R X4 Selected from C1-C8 alkyl groups substituted or unsubstituted by one or more substituents, or C1-C8 alkoxy groups substituted or unsubstituted by one or more substituents, wherein the substituents are selected from deuterium or halogens; In particular, At least one of R1, R2, R3, and R4 is either deuterium or is replaced by deuterium.

2. The novel deuterium-containing pyrazolone compound, tautomer, stereoisomer, prodrug, and pharmaceutically acceptable salt thereof as described in claim 1, having the structure of formula (III) and / or formula (IV): The substituents in formula (III) and / or formula (IV) are defined as defined in formula (I) and / or formula (II) as claimed in claim 1.

3. The novel deuterium-containing pyrazolone compounds, tautomers, stereoisomers, prodrugs, and pharmaceutically acceptable salts thereof as described in claims 1-2, wherein the compounds include, but are not limited to, the following compounds:

4. A pharmaceutical composition comprising any novel deuterium-containing pyrazolinone compound, tautomer, stereoisomer, prodrug, or pharmaceutically acceptable salt thereof as described in any one of claims 1 to 3.

5. Use of the novel deuterium-containing pyrazolone compound, tautomer, stereoisomer, prodrug, or pharmaceutically acceptable salt thereof, as described in any one of claims 1 to 3, in the preparation of a neuroprotective medicament.

6. Use of any novel deuterium-containing pyrazolone compound, tautomer, stereoisomer, prodrug, or pharmaceutically acceptable salt thereof, as described in any one of claims 1 to 3, in the preparation of a medicament for the prevention or treatment of cardiovascular and cerebrovascular diseases.

7. The use according to claim 5, wherein the neuroprotective drug is a drug for treating neuropathic diseases, wherein the neuropathic diseases are Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, Huntington's disease, multiple sclerosis, cerebellar ataxia, different types of spinocerebellar ataxia, spinal muscular atrophy, cerebral ischemia, and primary lateral sclerosis.

8. The use as described in claim 6, wherein the drug for preventing or treating cardiovascular and cerebrovascular diseases is a drug for treating or preventing cardiovascular and cerebrovascular diseases, wherein the cardiovascular and cerebrovascular diseases are hypertension, coronary heart disease, stroke, diabetic heart failure, heart failure, diastolic heart failure, systolic heart failure, postoperative volume overload, idiopathic edema, pulmonary hypertension, pulmonary arterial hypertension, and heart failure.

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