Preparation method and application of pyrazole compound

Abstract

The invention discloses a preparation method and application of a pyrazole compound, and relates to a compound with a novel structure and a composition of the compound, and the pyrazole compound shows better activity in vivo and does not show obvious toxicity. The compound provided by the invention can be developed into drugs for protecting multiple diseases such as nerve cells, cardiovascular diseases and the like. # imgabs0 #

Description

Technical Field

[0001] The present application relates to the field of pharmaceutical technology, and in particular to a pyrazole compound and a composition thereof. The compound and the composition thereof can be developed for use in treating diseases caused by oxidative stress and / or thrombosis caused by free radicals. Background Art

[0002] The human brain accounts for only 2.0% of body weight, but its oxygen consumption accounts for about a quarter of the total oxygen consumption of the whole body, making it the organ with the highest oxygen consumption per unit mass of human tissue at rest. Therefore, brain tissue produces more free radicals and peroxides, and is more susceptible to oxidative stress than other organs. Free radicals are the products of aerobic metabolism of cells, mainly superoxide anion free radicals, lipid oxygen free radicals, nitric oxide free radicals, hydroxyl free radicals, etc. Free radicals are highly active and can affect the functions of biological molecules. When nerve cells are affected by free radicals, they can affect the function of nerve cells, and severe cases can lead to neurodegenerative diseases. Common Alzheimer's disease (AD) and Parkinson's disease (PD) are all diseases caused by oxidative stress reactions. Oxidative stress also plays an important role in cerebrovascular diseases. When cerebral vascular endothelial cells are damaged, it can induce the formation of arteriosclerotic plaques, which in turn lead to acute cerebral vascular thrombosis, resulting in local insufficient or interrupted blood supply to the brain, and irreversible damage to the brain. Oxidative stress is also closely related to cardiovascular diseases. For example, common atherosclerosis is the main cause of cardiovascular diseases. Oxidative stress plays an important role in the pathogenesis of cardiovascular diseases such as myocardial ischemia, cardiac systolic dysfunction, and myocardial infarction.

[0003] Antioxidant drugs can effectively scavenge free radicals, protect nerve cells, prevent the occurrence of neurodegenerative diseases, and thus play a therapeutic role. Ligustrazine (TMP) is a natural product extracted from the traditional Chinese medicine Chuanxiong. It has the effects of inhibiting platelet aggregation and anti-thrombosis. However, Ligustrazine has poor lipid solubility and a short half-life. It must be administered frequently in clinical practice, which can easily lead to adverse events such as cumulative poisoning. Therefore, its clinical application is greatly limited. Edaravone is the first approved drug that can scavenge free radicals. It can inhibit the damage of free radicals to brain cells, vascular endothelial cells, and nerve cells.

[0004] In view of the limitations of ligustrazine in clinical use as a drug, it is necessary to structurally modify ligustrazine in order to obtain a more ideal clinical drug. Summary of the invention

[0005] The present inventors have developed a novel compound, which has obvious advantages over the marketed drug edaravone in treating diseases caused by oxidative stress and / or thrombosis caused by free radicals.

[0006] The present invention provides a pyrazole compound and a composition thereof which can be developed into drugs for protecting nerve cells, cardiovascular diseases and other diseases.

[0007]

[0008] including isomers, solvates, isotopic derivatives, oxides, and salts thereof;

[0009] in:

[0010] R0 is hydrogen,

[0011] Y1 and Y2 are each independently CH or N;

[0012] R1, R2, and R3 are each independently hydrogen, deuterium, C 1-6 Alkyl or deuterated C 1-6 alkyl;

[0013] R4 is hydrogen, deuterium, C 1-6 Alkyl, C 1-6 Alkylamino, or C 1-6 Alkoxy;

[0014] R5 is hydrogen, deuterium, C 1-6 Alkyl, C 1-6 Alkylamino, C 1-6 Alkyl alcohol, or

[0015] Wherein, m is 1, 2 or 3;

[0016] n is 1, 2 or 3;

[0017] When R5 is When R0 is hydrogen, When R5 is hydrogen, deuterium, C 1-6 Alkyl, C 1-6 Alkylamino, C 1-6 In the case of alkoxy, R0 is

[0018]

[0019] R6 is hydrogen, deuterium, C 1-6 Alkyl, C 1-6 Alkylamino, C 1-6 Alkoxy;

[0020] R7 is C 1-6 Alkylamino, C6-12 Alkyl, C 6-12 Alkoxy;

[0021] R8 and R9 are each independently hydrogen, C 1-6 Alkylamino, C 6-12 Alkyl, C 6-12 Alkoxy, and R8 and R9 are not hydrogen at the same time;

[0022] R 10 C 1-6 Alkylamino, C 1-6 Alkyl carboxylic acid, or -(CR 12 R 13 ) p NR 14 CO(CR 15 R 16 ) q R 17 ;

[0023] R 11 C 1-6 Alkyl, or -CO(CR 18 R 19 ) y R 20 ;

[0024] Wherein, p is 2 or 3;

[0025] q is 1, 2, or 3;

[0026] y is 1, 2, or 3;

[0027] R 12 , R 13 are independently hydrogen, C 1-6 alkyl;

[0028] R 14 For hydrogen, C 1-6 alkyl;

[0029] R 15 , R 16 are independently hydrogen, amino, C 1-6 alkyl;

[0030] R 17 C 1-6 Alkylamino, C 6-12 Alkyl, C 6-12 Alkoxy, C 1-6 Alkyl carboxylic acids;

[0031] R 18 , R 19 are independently hydrogen, amino, C1-6 alkyl;

[0032] R 20 C 1-6 Alkylamino, C 6-12 Alkyl, C 6-12 Alkoxy, C 1-6 Alkyl carboxylic acid.

[0033] In some embodiments, the present invention provides a compound as described in formula (II):

[0034] The substituents in formula II are as defined in formula I.

[0035] In some embodiments, the present invention provides a compound as described in formula (III):

[0036] The substituents in formula III are as defined in formula I.

[0037] In some embodiments, the present invention provides a compound as described in formula (IV):

[0038] The substituents in formula IV are as defined in formula I.

[0039] In some embodiments, the present invention provides a compound as described in formula (V):

[0040] The substituents in formula V are as defined in formula I.

[0041] In some embodiments, the present invention provides a compound as described in formula (VI):

[0042]

[0043] The substituents in formula VI are as defined in formula I.

[0044] In some embodiments, the present invention provides a pyrazole compound of formula (I) to formula (VI), isomers, solvates, isotopic derivatives, oxides, and salts thereof.

[0045] In some embodiments, in the above formula (I)-formula (VI), Y1 is selected from CH, or N.

[0046] In some embodiments, in the above formula (I)-formula (VI), Y2 is selected from CH, or N.

[0047] In some embodiments, in the above formula (I) to formula (VI), R1 is selected from hydrogen, deuterium, C 1-6 Alkyl or deuterated C 1-6 Alkyl; preferably, R1 is hydrogen or deuterium.

[0048] In some embodiments, in the above formula (I) to formula (VI), R2 is selected from hydrogen, deuterium, C 1-6 Alkyl or deuterated C 1-6 Alkyl; preferably, R1 is hydrogen or deuterium.

[0049] In some embodiments, in the above formula (I) to formula (VI), R3 is selected from hydrogen, deuterium, C 1-6 Alkyl or deuterated C 1-6 Alkyl; preferably, R1 is hydrogen or deuterium.

[0050] In some embodiments, in the above formula (I) to formula (VI), R4 is hydrogen, deuterium, C 1-6 Alkyl, C 1-6 Alkylamino, or C 1-6 Alkoxy; preferably, R4 is hydrogen, deuterium, methyl, ethyl, or isopropyl.

[0051] In some embodiments, in the above formula (I) to formula (IV), R5 is hydrogen, deuterium, C 1-6 Alkyl, C 1-6 Alkylamino, C 1-6 Alkyl alcohol, or Preferably, R5 is hydrogen, deuterium, methyl, ethyl, isopropyl, methyl alcohol, or

[0052] Wherein, the above m is 1, 2 or 3;

[0053] n is 1, 2 or 3; preferably, m is 1, and n is 2 or 3;

[0054] R 11 Selected from C 1-6 Alkyl, or -CO(CR 18 R 19 ) y R 20 ;

[0055] Wherein, the above y is 1, 2, or 3; preferably, y is 1;

[0056] R 18 , R 19 are independently hydrogen, amino, C 1-6 Alkyl; preferably, R 18 is hydrogen, R 19 It is amino group;

[0057] R20 C 1-6 Alkylamino, C 6-12 Alkyl, C 6-12 Alkoxy, C 1-6 Alkyl carboxylic acid; preferably, R 20 It is methyl, ethyl or isopropyl.

[0058] In some embodiments, in the above formula (I), formula (V), and formula (VI), R5 is in,

[0059] The above m is 1, 2 or 3;

[0060] n is 1, 2 or 3;

[0061] Preferably, m is 1, and n is 2 or 3;

[0062] R 11 Selected from C 1-6 Alkyl, or -CO(CR 18 R 19 ) y R 20 ;

[0063] Wherein, the above y is 1, 2, or 3; preferably, y is 1;

[0064] R 18 , R 19 are independently hydrogen, amino, C 1-6 Alkyl; preferably, R 18 is hydrogen, R 19 It is amino group;

[0065] R 20 C 1-6 Alkylamino, C 6-12 Alkyl, C 6-12 Alkoxy, C 1-6 Alkyl carboxylic acid; preferably, R 20 It is methyl, ethyl or isopropyl.

[0066] In some embodiments, in the above formula (I) to formula (VI), R6 is hydrogen, deuterium, C 1-6 Alkyl, C 1-6 Alkylamino, C 1-6 Alkoxy; preferably, R6 is hydrogen, deuterium, methyl, ethyl, or isopropyl.

[0067] In some embodiments, in the above formula (I) and formula (V), R0 is hydrogen, in,

[0068] The above R7 is C 1-6 Alkylamino, C6-12 Alkyl, C 6-12 Alkoxy; preferably, R7 is

[0069] R8 and R9 are each independently hydrogen, C 1-6 Alkylamino, C 6-12 Alkyl, C 6-12 Alkoxy, and R8 and R9 are not hydrogen at the same time; preferably, R8 is hydrogen, and R9 is

[0070] R 10 C 1-6 Alkylamino, C 1-6 Alkyl carboxylic acid, or -(CR 12 R 13 ) p NR 14 CO(CR 15 R 16 ) q R 17 ,

[0071] Wherein, p is 2 or 3;

[0072] q is 1, 2, or 3;

[0073] Preferably, p is 3;

[0074] q is 1;

[0075] R 12 , R 13 are independently hydrogen, C 1-6 Alkyl; preferably, R 12 and R 13 is hydrogen;

[0076] R 14 For hydrogen, C 1-6 Alkyl; preferably, R 14 is hydrogen;

[0077] R 15 , R 16 are independently hydrogen, amino, C 1-6 Alkyl; preferably, R 15 is hydrogen, R 16 It is amino group;

[0078] R 17 C 1-6 Alkylamino, C 6-12 Alkyl, C 6-12 Alkoxy, C 1-6Alkyl carboxylic acid; preferably, R 17 It is isopropyl.

[0079] In some embodiments, in the above formula (I) and formula (II), R0 is Wherein, the above R7 is C 1-6 Alkylamino, C 6-12 Alkyl, C 6-12 Alkoxy; preferably, R7 is

[0080] In some embodiments, in the above formula (I) and formula (III), R0 is

[0081] wherein the above R8 and R9 are each independently hydrogen, C 1-6 Alkylamino, C 6-12 Alkyl, C 6-12 Alkoxy, and R8 and R9 are not hydrogen at the same time; preferably, R8 is hydrogen, and R9 is

[0082] In some embodiments, in the above formula (I) and formula (IV), R0 is

[0083] in,

[0084] The above R 10 C 1-6 Alkylamino, C 1-6 Alkyl carboxylic acid, or -(CR 12 R 13 ) p NR 14 CO(CR 15 R 16 ) q R 17 ,

[0085] Wherein, p is 2 or 3;

[0086] q is 1, 2, or 3;

[0087] Preferably, p is 3;

[0088] q is 1;

[0089] R 12 , R 13 are independently hydrogen, C 1-6 Alkyl; preferably, R 12 and R 13 is hydrogen;

[0090] R 14 For hydrogen, C 1-6Alkyl; preferably, R 14 is hydrogen;

[0091] R 15 , R 16 are independently hydrogen, amino, C 1-6 Alkyl; preferably, R 15 is hydrogen, R 16 It is amino group;

[0092] R 17 C 1-6 Alkylamino, C 6-12 Alkyl, C 6-12 Alkoxy, C 1-6 Alkyl carboxylic acid; preferably, R 17 It is isopropyl.

[0093] The compound has the following structure:

[0094]

[0095]

[0096]

[0097] The compound and its salts, solvates, isomers and polymorphs are used in preparing medicines for treating related diseases caused by free radicals.

[0098] The pharmaceutical composition is characterized in that it comprises a therapeutically effective amount of a compound described in any one of the above structures and a salt thereof and a pharmaceutically acceptable carrier, adjuvant, vehicle and a combination thereof.

[0099] The use is characterized in that it can be used to treat diseases caused by oxidative stress and / or thrombosis caused by free radicals.

[0100] The use is characterized in that the diseases caused by oxidative stress and / or thrombosis caused by free radicals include arteriosclerosis, Alzheimer's disease, Parkinson's disease, multiple sclerosis, or amyotrophic lateral sclerosis, heart failure, heart disease, cerebral stroke, myocardial ischemia and ischemia-reperfusion injury, myocardial infarction, coronary heart disease, or heart failure.

[0101] The method for treating a patient suffering from the disease is characterized by comprising administering the compound or the pharmaceutical composition to the subject.

[0102] The compound of the present invention has a better neuroprotective function, a significantly increased half-life, and an effectively improved free radical scavenging rate.

[0103] The compounds and salts thereof of the present invention, pharmaceutically acceptable carriers, adjuvants, vehicles and compositions thereof exhibit better stability during storage and are more tolerant to external environments.

[0104] In the present invention, for a more detailed understanding of the present invention, each term has the following definitions.

[0105] The terms "a", "an" or similar terms herein refer to objects including one or more numbers, or mixtures thereof, and are not limited to one or one.

[0106] The term "independently" as used herein means that with more than one variable, each instance of a substituent is selected from the available definitions of the variable independently of the other selected defined variables. Thus, each substituent may be the same as or different from the other substituents.

[0107] The term "or" or similar terms herein refers to any one member or object within a specific scope, and a combination of multiple members or objects.

[0108] The term "including" herein means including but not limited to, and does not limit or exclude, for example, other classified species or compositions.

[0109] In this article, the term “C 1-6 The term "alkyl" refers to a straight chain, branched or unbranched saturated or unsaturated aliphatic hydrocarbon group containing up to 6 carbon atoms. Specific examples of alkyl groups include methyl, ethyl, isopropyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 2,2-dimethylpropyl, and the like.

[0110] In this article, the term “C 1-6 "Alkoxy" means an alkyl group containing up to 6 carbon atoms with an -O- or -OH group inserted in any reasonable position. The group may be straight chain, branched or unbranched. Specific examples include methoxy, ethoxy, isopropoxy, tert-butoxy, isopentyl, cyclopentyloxy, 2-methoxybutyl, 2-ethoxypropyl, 2-hydroxybutyl, and the like.

[0111] In this article, the term “C 1-6 "Alkylamino" means an alkyl group containing up to 6 carbon atoms with -N-, -NH, or -NH2 groups inserted in any reasonable position. The group can be straight chain, branched or unbranched. Specific examples are methylamino, ethylamino, isopropylamino, tert-butylamino, cyclopentylamino, 2-methylaminobutyl, 2-ethylaminopropyl, 2-aminobutyl, etc.

[0112] In this article, the term “C 1-6"Sulfonylalkylamino" means a group in which a sulfonylalkyl group containing up to 6 carbon atoms is inserted with -N-, -NH, or -NH2 groups at any reasonable position. The group may be straight chain, branched or unbranched. Specific examples are sulfonylmethylamino, sulfonylethylamino, sulfonylisopropylamino, 2-methylaminobutylsulfonyl, 2-ethylaminopropylsulfonyl, 2-aminobutylsulfonyl, and the like.

[0113] When the compound of the present invention has one or more chiral centers, all enantiomers and diastereomers, or the use of mixtures thereof, and the use of active ingredient pharmaceutical compositions are included. When the compound has double bonds, all configurational isomers (such as cis and trans isomers), or the use of mixtures thereof, and the use of active ingredient pharmaceutical compositions are included.

[0114] The compounds of the present invention also include their tautomers, such as positive ion shift tautomerism, negative ion shift tautomerism, bimolecular proton shift isomerism, intra-ring tautomerism, ring-chain tautomerism, keto-enol tautomerism, amide-imidic acid isomerism, lactam-lactimide isomerism, etc.

[0115] The term "solvate" herein refers to a compound obtained by incorporating suitable solvent molecules into the crystal lattice of the compound of the present invention and its salt, and combining them through coordination bonds (such as solvated metal ions) or covalent bonds. Suitable solvents include acetic acid, ethanol, water, acetone, etc. When the suitable solvent molecule is water, it can be referred to as "hydrate".

[0116] The term "isotopic derivative" as used herein refers to a product obtained by replacing one or more isotopic atoms in a molecule. "Isotopes" as used herein refer to atoms with the same atomic number but different proton numbers, such as 2 H. 3 H. 17 O. 18 O. 18 F. 13 C. 15 N. 36 Cl, 32 P. 11 B, etc. Isotope-substituted compounds can be used to study drug metabolism and chemical reaction kinetics.

[0117] The compounds of the present invention contain basic nitrogen atoms (heterocyclic or aliphatic amino groups, etc.), which are easily oxidized to N-oxides by oxidants such as oxygen in the air and hydrogen peroxide to generate other compounds of the present invention. Therefore, the converted N-oxide derivatives are considered as part of the compounds of the present invention.

[0118] The term "salt" herein refers to a complex formed by the combination of a compound and a corresponding acid, the properties of which depend on the characteristics of the compound, such as an addition salt of the compound with an acid, for example, an inorganic acid salt such as hydrochloride, sulfate, hydrobromide, etc. Organic acid salts such as maleate, fumarate, acetate, propionate, malate, tartrate, malonate, succinate, citrate, cinnamate, mandelate, methanesulfonate, p-toluenesulfonate, salicylate, etc.

[0119] The term "salt" herein also refers to addition salts of the compound with a base, inorganic base salts such as sodium salts, potassium salts, ammonium salts, calcium salts, magnesium salts, etc. Salts of organic amines such as diethylamine salts, ethylenediamine salts, meglumine salts, tromethamine salts, arginine salts, lysine salts, histidine salts, piperidine salts, etc.

[0120] The term "treatment" as used herein refers to the act of inhibiting the progression of, or reversing the symptoms of, the applicable disorder or one or more symptoms, and also includes auxiliary treatments for symptoms.

[0121] The term "pharmaceutically acceptable carrier, adjuvant, vehicle and composition thereof" herein refers to diluents, starch, dextrin, sugars, mannitol, microcrystalline cellulose, oils, binders, wetting agents, distilled water, ethanol, binders, starch slurry, disintegrants, lubricants, glidants, etc.

[0122] The term "therapeutically effective amount" herein refers to the active ingredient being administered in the range of 0.01% to 75% (W / W%), and being administered to the desired individual in one or more dosage forms at appropriate time intervals every day, which can produce significant therapeutic results in vivo. In addition, the dosage can be changed by judging the severity of the disease, the age and weight of the subject, etc. to achieve the desired therapeutic effect.

[0123] The term "subject" as used herein refers to warm-blooded animals, such as humans, rats, guinea pigs, mice, gerbils, rabbits, dogs, pigs, sheep, monkeys, chickens, ducks, geese, cats, cows, horses, chimpanzees, and the like.

[0124] The compounds and compositions provided by the present invention can be prepared into pharmaceutical preparations and administered orally, intranasally, intraorally, on the skin, intravenously, and in a variety of ways to prevent or treat an individual in need of treatment. DETAILED DESCRIPTION

[0125] The present invention is further described below in conjunction with examples, but these examples do not limit the scope of the present invention. Unless otherwise specified, the experimental solvents or reagents used in the following examples were purchased from the market and used directly without further purification.

[0126] Example 1: Preparation and characteristics of compound M-2

[0127]

[0128] Synthesis of compound M-1: SM01 (5.0 g, 31.93 mmol) was dissolved in hydrazine hydrate (50 mL), and the temperature was raised to 110°C for 8 hours. After TLC detection, the reaction was completed, the system was cooled, filtered, washed with water, concentrated and dried to obtain 2.38 g of gray solid, with a yield of 49.0%.

[0129] 1 HNMR(400MHz)δ:8.44(brs,1H),5.01(brs,2H),2.68(s,6H),2.34(s,3H).

[0130] ESI-MS (+): m / z = 153.47.

[0131] Synthesis of compound M-2: Compound M-1 (2.38 g, 15.64 mmol) was dissolved in toluene, ethyl acetoacetate (3.05 g, 23.47 mmol) was added, and the temperature was raised to reflux for 6 hours after the addition was complete. TLC detected that the reaction was complete, cooled, washed with water, concentrated to remove the solvent, and then column chromatography was performed to obtain a light yellow solid: 0.85 g, yield: 25.0%,

[0132] 1 HNMR(400MHz)δ:10.81(s,1H),6.49(s,1H),2.77-2.70(d,6H),2.34(s,3H),2.31(s,3H).

[0133] ESI-MS (+): m / z = 219.44.

[0134] Example 2: Preparation and Characterization of Compound M-6

[0135]

[0136] Synthesis of compound M-3: Compound SM01 (5.0 g, 31.93 mmol) was dissolved in DCM, cooled to 0°C, and m-CPBA (80%, 8.27 g, 38.32 mmol) was slowly added. After the addition was completed, the temperature was naturally raised to room temperature for reaction for 8 hours. After TLC detection, the reaction was completed, and after filtering to remove the solid, the mother liquor was concentrated, and the concentrated residue was dissolved in water. The pH was adjusted to 8 by sodium hydroxide, and the aqueous phase was extracted with DCM. After separation, the organic phase was dried and concentrated to obtain a light yellow liquid: 5.6 g, which was directly used in the next step without further purification.

[0137] ESI-MS (+): m / z = 173.71.

[0138] Synthesis of compound M-4: Compound M-3 (5.6 g) was added to hydrazine hydrate, heated to 110°C, and reacted for 6 hours. After TLC detection, the reaction was completed, the system was cooled to room temperature, poured into water, extracted with DCM, and after separation, the organic phase was dried and concentrated to obtain an off-white solid: 2.68 g, two-step yield: 50%,

[0139] ESI-MS (+): m / z = 169.76.

[0140] Synthesis of compound M-5: Compound M-4 (2.68 g, 15.96 mmol) was dissolved in toluene, ethyl acetoacetate (3.1 g, 23.94 mmol) was added, and the temperature was raised to reflux for 6 hours after the addition was complete. TLC detected that the reaction was complete, cooled, washed with water, and concentrated to remove the solvent to obtain a yellow solid: 0.93 g, yield: 25%,

[0141] ESI-MS (+): m / z = 235.72.

[0142] Synthesis of compound M-6: Compound M-5 (0.93 g, 39.69 mmol) was dissolved in acetic anhydride and heated to 80°C for 4 hours. After the reaction was completed by TLC detection, the system was cooled to room temperature and poured into ice water. 50% sodium hydroxide aqueous solution was used to adjust the pH to 10, and then the system was heated to 80°C for 2 hours. After the reaction was completed by TLC detection, the system was cooled to room temperature, extracted with DCM, and after separation, the organic phase was dried and concentrated to obtain a light yellow oil: 0.4 g, yield: 43%,

[0143] 1 HNMR(400MHz)δ:11.85(s,1H),6.35(s,1H),5.41(s,1H),4.68(s,2H),2.68(s,3H),2.41(s,3H),2.29(s,3H).

[0144] ESI-MS (+): m / z = 235.54.

[0145] Example 3: Preparation and Characterization of Compound M-8

[0146]

[0147] Synthesis of compound M-8: SM02 (10.0 g, 62.43 mmol) was dissolved in ethanol, cooled to 0°C, and an ethanol solution of sodium hydroxide (2.50 g, 62.43 mmol) was slowly added. After the addition, the mixture was reacted for 1 hour. A large amount of solid appeared in the system. The filter cake was washed with ethyl acetate and dried to obtain a white solid: 9.5 g, yield: 89%.

[0148] Synthesis of compound M-8: M-7 (9.5 g, 55.82 mmol) and magnesium chloride (5.8 g, 61.40 mmol) were mixed in acetonitrile, and triethylamine (5.65 g, 55.82 mmol) was added. After the addition, the mixture was stirred at 10°C for 3 hours, then cooled to 0°C, and SM03 (6.8 g, 83.73 mmol) was added dropwise. After the addition, the mixture was naturally warmed to room temperature and reacted for 12 hours. The solvent was then removed by concentration, and the concentrated residue was dissolved in toluene. The pH was adjusted to 3 with 1N hydrochloric acid, and the liquid was separated. The organic phase was concentrated to obtain a colorless liquid: 8.9 g, yield: 90%, and it was set aside for later use.

[0149] Example 4: Preparation and Characterization of Compound M-9

[0150]

[0151] Synthesis of compound M-9: Compound M-1 (1.0 g, 6.57 mmol) was dissolved in toluene, and M-8 (1.4 g, 7.9 mmol) was added. After the addition, the temperature was raised to reflux for 8 hours. TLC detected that the reaction was complete, and the system was cooled to room temperature, washed with water, and the organic phase was dried and concentrated to obtain a residue. Column chromatography obtained an off-white solid: 0.54 g, yield: 37%,

[0152] 1 HNMR(400MHz)δ:11.67(s,1H),6.39(s,1H),2.75-2.71(d,6H),2.45(s,3H).

[0153] ESI-MS (+): m / z = 222.22.

[0154] Example 5: Preparation and Characterization of Compound M-11

[0155]

[0156] Synthesis of compound M-10: Compound M-9 (1.0 g, 4.52 mmol) was dissolved in DCM (50 mL), and the temperature was lowered to 5 °C and then m-CPBA (1.2 g, 5.42 mmol) was slowly added. After the addition, the temperature was naturally raised to room temperature and the reaction was carried out for 24 hours. During the reaction, a white solid was precipitated. The reaction was completed by TLC detection. The system was filtered, and the filter cake was washed with DCM. The filtrates were combined and concentrated to obtain the crude product M-18, which was directly used in the next step without purification.

[0157] Synthesis of compound M-11: M-10 (1.0 g) obtained by concentration in the previous step was dissolved in acetic anhydride (20 mL), and heated to 80°C for 4 hours. The reaction was complete by TLC detection. After the system cooled to room temperature, it was poured into an iced 50% aqueous sodium hydroxide solution, and then heated to 80°C for 2 hours. The reaction was complete by TLC detection, and then the system was cooled to room temperature, extracted with DCM, and the organic phase was dried and concentrated after separation to obtain a yellow oil: 0.53 g, yield: 49.53%,

[0158] 1 HNMR(400MHz)δ:11.85(s,1H),6.35(s,1H),5.41(s,1H),4.68(s,2H),2.68(s,3H),2.41(s,3H).

[0159] ESI-MS (+): m / z = 238.38.

[0160] Example 6: Preparation and Characterization of Compound M-12

[0161]

[0162] Synthesis of compound M-12: The synthesis scheme is shown in Example 4. Compound M-1 was replaced with compound SM04 (1.0 g, 9.2 mmol) to obtain a light yellow oil: 0.51 g, yield: 34%,

[0163] 1 HNMR(400MHz)δ:11.56(ss,1H),7.92-7.17(m,5H),6.70(s,1H).

[0164] Example 7: Preparation and Characterization of Compound M-14

[0165]

[0166] Synthesis of compound M-13: SM05 (10.0 g, 79.9 mmol) was dissolved in THF, cooled to 0°C, 50% sodium hydroxide (9.6 g, 0.24 mmol) aqueous solution was slowly dripped into the system, and then Boc2O (26.2 g, 119.8 mmol) was slowly dripped into the system, and the temperature was naturally raised to room temperature after the addition, and the reaction was carried out for 12 hours. TLC detected that the reaction was complete, the system was cooled to 5°C, 1N hydrochloric acid was used to adjust the pH to 3, and then DCM was extracted. After separation, the organic phase was dried and concentrated to obtain a colorless liquid: 15 g, yield: 83.3%, MS: m / z=224.7 (M-1);

[0167] Synthesis of compound M-14: Compound M-6 (1.0 g, 4.27 mmol) was dissolved in DCM, triethylamine (0.86 g, 8.54 mmol) was added, and M-13 (1.4 g, 6.41 mmol) was slowly added dropwise at room temperature. After the addition, the mixture was stirred at room temperature for 72 hours. TLC detected that the reaction was complete, and the reaction system was washed with water, dried and concentrated. Column chromatography gave a colorless oil: 1.4 g, yield: 74.47%,

[0168] 1 HNMR(400MHz)δ:12.01(brs,1H),6.84(brs,1H),6.14(s,1H),4.81(s,2H),3 .40(t,2H),3.30(t,2H),2.75(s,3H),2.44(s,3H),2.30(s,3H),1.42(s,9H).

[0169] ESI-MS (+): m / z = 442.51.

[0170] Example 8: Preparation and characteristics of DSC194-01

[0171]

[0172] Synthesis of compound M-15: Compound SM06 (100 mg, 0.65 mmol) and triethylamine (98 mg, 0.97 mmol) were dissolved in DCM, the temperature was controlled at 0°C, and a DCM solution of triphosgene (212 mg, 0.72 mmol) was added dropwise. After the addition, the mixture was reacted at room temperature for 2 hours, and then the solvent was directly concentrated to obtain a white solid, which was directly used in the next step without purification.

[0173] Synthesis of DSC194-01: The concentrated residue of M-15 was redissolved in DCM and added dropwise to a DCM solution of M-2 (127 mg, 0.59 mmol) at 0°C. After the addition, DBU (179 mg, 1.18 mmol) was added to the system. After the addition, the temperature was naturally raised to room temperature for 24 hours. After TLC detection, the reaction was complete, and the system was poured into water, extracted with DCM, and the organic phase was dried and concentrated after separation. The concentrated residue was purified by column chromatography to obtain 101.96 mg of compound DSC193-01, with a yield of 43.75%.

[0174] 1 HNMR(400MHz)δ:6.43(s,1H),4.35(m,1H),2.79(s,3H),2.77(s,3H),2.39(s,3H),2.30(s,3H),1.95`1.30(m,7H),0.90(s,3H),0.87(s,6H).

[0175] ESI-MS (+): m / z = 399.63.

[0176] Example 9: Preparation and characteristics of DSC194-06

[0177]

[0178] Synthesis of compound M-16: Compound M-2 (1.0 g, 4.58 mmol) was dissolved in toluene, triethylamine (0.56 g, 5.50 mmol) was added, cooled to 0°C, and then phosphorus oxychloride (0.77 g, 5.04 mmol) was slowly added dropwise. After addition, the mixture was reacted at 5°C for 3 hours, and then slowly added dropwise to ice water. DCM was extracted, and the organic phase was dried and concentrated after separation to obtain a yellow solid: 0.73 g, yield: 48.67%.

[0179] 1 HNMR(400MHz)δ:6.52(s,1H),3.01(s,3H),2.99(s,3H),2.65(s,3H),2.56(s,3H).

[0180] ESI-MS (+): m / z = 336.22.

[0181] Synthesis of DSC194-06: M-16 (0.73 g, 2.18 mmol) was dissolved in acetonitrile, and borneol (0.44 g, 2.83 mmol) and potassium carbonate (0.9 g, 6.53 mmol) were added. After the addition, the system was heated to reflux and reacted for 6 hours. Then the system was cooled to 0°C, 1N hydrochloric acid was slowly added dropwise, and the pH was adjusted to 3-4. After completion, the solvent was directly concentrated and the concentrated residue was prepared by liquid phase to obtain DSC194-06: 0.31 g, yield: 32.77%,

[0182] 1 HNMR(400MHz)δ:6.49(s,1H),3.35(m,1H),2.87(s,3H),2.85(s,3H),2.55(s,3H),2.27(s.3H),2.04-1.42(m,7H),0.90(s,3H),0.87(s,6H).

[0183] ESI-MS (+): m / z = 435.71.

[0184] Example 10: Preparation and characteristics of DSC194-11

[0185]

[0186] Synthesis of DSC194-11: SM07 (1.0 g, 5.74 mmol) and fresh concentrated residue M-15 (2.5 g) were dissolved in DCM (50 mL) under ice-water bath, and DBU (1.31 g, 8.61 mmol) was added under stirring. After addition, the temperature was naturally raised to room temperature for 24 hours. After TLC detection, the reaction was complete, and the system was poured into water, extracted with DCM, and the organic phase was dried and concentrated after separation. The concentrated residue was purified by column chromatography to obtain 0.31 g of white solid DSC194-11, with a yield of 15.27%.

[0187] 1 HNMR(400MHz)δ:7.81(d,2H),7.54-7.50(m,3H),6.70(s,1H),4.43(m,1H),2.30(s,3H),1.95-1.30(m,7H),0.90(s,3H),0.87(s,6H).

[0188] ESI-MS (+): m / z = 355.34.

[0189] Example 11: Preparation and characteristics of DSC194-16

[0190]

[0191] Synthesis of compound M-17: SM07 (1.0 g, 5.74 mmol) was dissolved in DCM, TEA (0.64 g, 6.31 mmol) was added, phosphorus oxychloride (1.32 g, 8.62 mmol) was slowly added dropwise after cooling to 0°C, and the reaction temperature was maintained and stirred for 4 hours after the addition. The reaction was completed by TLC detection, the solvent was concentrated and removed, and the residue was used directly in the next step without purification;

[0192] Synthesis of DSC194-16: The concentrated residual M-17 was dissolved in DCM, and SM06 (1.33 g, 8.62 mmol) was added. After the addition, the mixture was cooled to 5°C, and then triethylamine (2.91 g, 28.7 mmol) was added. After the addition, the temperature was maintained for 12 hours, and then the temperature was raised to 40°C and refluxed for 6 hours. The reaction was completed by HPLC detection, and the system was cooled to room temperature and poured into ice water. The pH was adjusted to 7 with sodium carbonate, and the solvent was removed after concentration. The preparative liquid phase separation was obtained to obtain a white solid: 0.75 g, with a yield of 33.48%.

[0193] 1 HNMR(400MHz)δ:7.56-7.17(m,5H),6.70(s,1H),3.37(m,1H),2.15-1.28(m,7H),0.89(s,3H),0.85(s,6H).

[0194] ESI-MS (+): m / z = 391.18.

[0195] Example 12: Preparation and characteristics of DSC194-19

[0196]

[0197] Synthesis of DSC194-19: M-12 (1.0 g, 5.64 mmol) and fresh concentrated residue M-15 (2.5 g) were dissolved in DCM (50 mL) under ice-water bath, and DBU (1.31 g, 8.61 mmol) was added under stirring. After addition, the temperature was naturally raised to room temperature for 24 hours. After TLC detection, the reaction was complete, and the system was poured into water, extracted with DCM, and the organic phase was dried and concentrated after separation. The concentrated residue was purified by column chromatography to obtain 0.47 g of white solid DSC194-19, with a yield of 23.27%.

[0198] 1 HNMR(400MHz)δ:7.79(d,2H),7.54-7.50(m,3H),6.70(s,1H),4.43(m,1H),1.95-1.30(m,7H),0.90(s,3H),0.87(s,6H).

[0199] ESI-MS (+): m / z = 358.54.

[0200] Example 13: Preparation and characteristics of DSC194-21

[0201]

[0202] Synthesis of compound M-18: M-5 (1.0 g, 4.27 mmol) was dissolved in DCM, cooled to 0°C, triethylamine (1.30 g, 12.9 mmol) was added, and then M-13 (1.44 g, 6.41 mmol) was added. After the addition, the temperature was naturally raised to room temperature and reacted for 72 hours. TLC detected that the reaction was complete, and the system was poured into ice water, extracted with DCM, and the organic phase was dried and concentrated after separation. The concentrated residue was used directly in the next step without treatment;

[0203] Synthesis of compound M-19: The concentrated residue of M-18 was dissolved in acetic anhydride and heated to 110°C for 2 hours. After TLC detection, the reaction was completed. After the system was cooled to room temperature, it was slowly dripped into ice water, extracted with DCM, and the organic phase was concentrated after separation. The concentrated residue was dissolved in THF, and 50% sodium hydroxide aqueous solution was added at room temperature, and then the temperature was raised to 90°C for 2 hours. After TLC detection, the reaction was completed, the reaction system was extracted with DCM, and after separation, the organic phase was dried and concentrated, and used directly in the next step without purification;

[0204] Synthesis of DSC194-21: The concentrated residue of M-19 was dissolved in ethanol, and ethanolic hydrochloric acid was added at room temperature. After the addition, the mixture was stirred for 6 hours. After the liquid phase detection, the reaction was completed, and the system was directly concentrated to remove the solvent. Then, an off-white oily substance was obtained by preparative liquid phase: 0.32 g, and the three-step yield was 21.92%.

[0205] 1 HNMR(400MHz)δ:6.38(s,1H),4.79(s,2H),3.30(t,2H),3.10(t,2H),2.77(s,3H),2.41(s,3H),2.31(s,3H).

[0206] ESI-MS (+): m / z = 342.37.

[0207] Example 14: Preparation and Characterization of DSC194-25

[0208]

[0209] Synthesis of compound M-20: SM07 (1.0 g, 5.75 mmol) and M-13 (1.29 g, 6.32 mmol) were mixed in toluene, p-toluenesulfonic acid (0.1 g) was added, and the temperature was raised to 80°C for 5 hours. The reaction was completed by TLC detection, the system was cooled to room temperature and concentrated, and then the residue was dissolved in EA, diluted with water, and then separated. After separation, the organic phase was washed with saturated brine, dried and concentrated to obtain crude M-21, which was directly used in the next step without separation;

[0210] Synthesis of DSC194-25: The concentrated residue of M-20 was dissolved in THF, TFA (5 mL) was added, and stirred at room temperature for 16 hours. TLC detected that the reaction was complete, and the system was directly concentrated and then separated by preparative liquid phase to obtain a solid: 0.23 g, yield: 14.20% (two steps),

[0211] 1 HNMR(400MHz)δ:7.56-7.13(m,5H),6.31(s,1H),3.29(t,2H),3.03(t,2H),2.30(s,3H).

[0212] ESI-MS (+): m / z = 282.43.

[0213] Example 15: Preparation and Characterization of DSC194-33

[0214]

[0215] Synthesis of compound M-21: M-5 (1.0 g, 4.27 mmol) and SM08 (0.79 g, 5.12 mmol) were mixed in toluene (50 mL), p-toluenesulfonic acid monohydrate (0.1 g) was added, and the temperature was raised to 100 ° C for 24 hours. The reaction was completed by HPLC, and the system was cooled to room temperature, concentrated, and the solvent was removed to obtain crude M-21, MS: m / z = 369.41 (M-1), which was used directly in the next step without purification;

[0216] Synthesis of DSC194-33: Dissolve the residual M-21 in acetic anhydride (20 g) and heat to 100°C for 4 hours. After HPLC detection, the reaction is complete. Cool the system to room temperature and pour it into an iced 50% sodium hydroxide aqueous solution, then heat to 80°C for 2 hours. After HLPC detection, the reaction is complete. Then cool the system to room temperature, adjust the pH to 3-4 with 2M HCL, concentrate to remove all solvents, and prepare the residue by preparative liquid phase to obtain a white solid: 0.27 g, yield: 17.09%,

[0217] 1 HNMR(400MHz)δ:12.01(brs,1H),6.50(s,1H),5.40(s,1H),4.83(d,2H),3.30(t,2H),2.81(s,3H),2.54(t,2H),2.47(s,3H),2.30(s,3H).

[0218] ESI-MS (-): m / z = 369.31.

[0219] Example 16: Preparation and Characterization of DSC194-37

[0220]

[0221] Synthesis of DSC194-37: SM07 (1.0 g, 5.74 mmol) and SM08 (0.93 g, 6.89 mmol) were mixed in toluene (50 mL), p-toluenesulfonic acid monohydrate (0.1 g) was added, and the temperature was raised to 100°C for 24 hours. The reaction was completed by HPLC, and the solvent was concentrated and removed. A white solid was obtained by preparative liquid phase: 0.53 g, yield: 29.78%,

[0222] 1 HNMR(400MHz)δ:12.01(brs,1H),7.80(d,2H),7.55-7.50(m,3H),6.70(s,1H),3.30(t,2H),2.49(t,2H),2.33(s,3H).

[0223] ESI-MS (-): m / z = 309.17.

[0224] Example 17: Preparation and Characterization of DSC194-41

[0225]

[0226] Synthesis of compound M-23: Freshly prepared M-13 (1.65 g, 5.12 mmol) and M-5 (1.0 g, 4.27 mmol) were mixed in DMF (20 mL) under water bath cooling, triethylamine (0.65 g, 6.40 mmol) was added under stirring, and stirred at room temperature for 16 hours. The reaction was completed by HPLC detection. The system was poured into ice water, extracted with DCM, and the organic phase after separation was dried over anhydrous sodium sulfate, and concentrated to remove the solvent to obtain crude M-24, which was used directly in the next step without purification;

[0227] Synthesis of DSC194-41: M-23 (residue from the previous step) was dissolved in DCM (50 mL), trifluoroacetic anhydride (1.79 g, 8.54 mmol) was added, and the mixture was stirred at room temperature for 24 hours. The reaction was completed by HPLC. The system was poured into ice water, separated, and the aqueous phase was extracted with DCM and separated. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. The concentrated residue was dissolved in THF (50 mL), 5 mL of 5M lithium hydroxide aqueous solution was added, and the mixture was stirred for 1 hour. The reaction was completed by HPLC. After adjusting the pH to 7 with 1M hydrochloric acid, the mixture was dried over anhydrous sodium sulfate and concentrated to remove the solvent. The prepared liquid phase was prepared to obtain a white solid: 0.32 g, yield: 18.08%;

[0228] 1 HNMR(400MHz)δ:6.39(s,1H),4,79(s,2H),4.40(m,1H),2.70(s,3H),2.37(s,3H),2.30(s,3H),1.94-1.20(m,7),0.88(s,3H),0.85(s,6H).

[0229] MS (+): m / z = 418.52.

[0230] The following compounds were prepared using a similar synthetic method:

[0231]

[0232]

[0233]

[0234] Example 18: Testing of the protective effect of compounds on glutamate-induced neuronal cell damage

[0235] Take the cerebral cortex of a newborn rat within 24 hours, place it in a culture dish, use D-Hank's solution to remove the membrane, and treat the damage. Transfer the treated rat cerebral cortex tissue to a conical flask, add 0.12% trypsin solution (Trypsin Solution), mix well and place in a 37°C incubator for 10 minutes. Then add serum-containing culture medium to terminate the enzyme reaction, place the cell suspension in a centrifuge, centrifuge at 1600r / min for 10 minutes, discard the supernatant, add 3mL of neuronal cell culture medium (Neurobasal Medium, GIBCO, 21103049), mix well and filter with a 200-mesh sieve, and count the cells in the supernatant. Then, use 1×10 4 The cells were inoculated into a 96-well plate at a density of 100 μL / well and incubated in a 37°C incubator for 72 hours. The volume of each well was fixed to 200 μL, and the corresponding compound was administered to each well to maintain a final concentration of 1.0 μM. Edaravone and DSC194-00 were positive control drugs. A normal control group and a glutamate injury group were also set up. Three parallel wells were set up for each drug concentration. After 30 minutes, Glu (5 μL, 200 μM) and Gly (5 μL, 200 μM) were administered to each well again. The cell state was observed during this period. The medium was replaced in full after 30 minutes. The culture medium (extracellular) was collected after being placed in the incubator for 8 hours. After washing twice with PBS solution, 220 μL of double distilled water was added, and then the cells were repeatedly frozen and thawed 3 times at -78°C. Then the culture medium (intracellular) was collected and the LDH leakage rate was determined according to the standard operating method. The absorbance OD value of each well at 440 nm was measured on an ELISA reader.

[0236] The leakage rate was calculated according to the following formula 1:

[0237] Formula 1: Leakage rate = extracellular OD value / (extracellular OD value + intracellular OD value) × 100%

[0238] Then the inhibition rate was calculated according to the following formula 2:

[0239] Formula 2: Inhibition rate (%) = (leakage rate of glutamate group - leakage rate of compound group) / (leakage rate of glutamate group - leakage rate of blank control group) × 100%

[0240] The calculation results are shown in Table 1:

[0241] Table 1: Average inhibition rate of neuronal cell damage by compounds at a concentration of 1.0 μM

[0242]

[0243]

[0244] Note: *DSC194-00 structural formula is: The structural formula of Edaravone is:

[0245] The results showed that the compounds DSC194-01, DSC194-06, DSC194-11, DSC194-16, DSC194-19, DSC194-21, DSC194-25, DSC194-33, DSC194-37, DSC194-40 and DSC194-41 of the present invention had significant inhibitory effects on glutamate-induced neuronal cell damage, and the inhibition rates of cell damage were all above 70%, which were higher than those of the positive controls edaravone (48.4%) and DSC194-00 (49. 3%), and the inhibition rates of compounds DSC194-01, DSC194-06, DSC194-11, DSC194-16, DSC194-19, DSC194-21, DSC194-25, DSC194-33, DSC194-37, DSC194-40 and DSC194-41 on glutamate-induced neuronal cell damage were more than 1.4 times that of edaravone and DSC194-00, respectively, showing good neuronal cell protective effects in vitro.

[0246] Example 10: Effects of compounds on LPS-induced zebrafish oxidation model

[0247] Normal zebrafish embryos were randomly selected and placed in a 12-well plate at a density of 25 per well. The blank control group was only added with an equal volume of buffer solution (5.0 mM NaCl, 0.17 mM KCl, 0.33 mM CaCl2, 0.33 mM MgSO4), which were set as: blank control group, model group, DSC194-01 group, DSC194-06 group, DSC194-11 group, DSC194-16 group, DSC194-19 group, DSC194-21 group, DSC194-25 group, DSC194-33 group, DSC194-37 group, DSC194-40 group, DSC194-41 group, edaravone group, DSC194-00 group, and three replicate wells were set for each group. After absorbing the water in each well, an equal amount of buffer solution was added to each well. Then, the corresponding compound solution (containing 0.1% DMSO for dissolution) was added to each well, and the final concentration of the compound was 1.0 μM. The blank control group and the model group only added an equal volume of buffer solution containing 0.1% DMSO. After 1 hour, 0.5 mL of LPS solution (final concentration of 10.0 μg / mL) was added to each group, and then placed in a 27°C incubator for further incubation for 24 hours. The experimental solution in each well was transferred to a new well plate, the original buffer solution was discarded, and a fresh equal volume of buffer solution was added. The wells were continued to be placed in a 37°C incubator for further incubation until 3 dpf, and then the egg membrane of each well was removed, and 5.0 ml (20 μg / ml) of fluorescent probe (DCHF-DA) was added to each well to detect the ROS content, and the embryos were incubated in the dark for 1 hour. The embryos were then washed three times with buffer solution, anesthetized with tricaine, and the fluorescence intensity was analyzed using ImageJ. The ROS inhibition rate was calculated according to the following formula 3:

[0248] Formula 3: Inhibition rate (%) = [1-(relative fluorescence intensity of the drug group-relative fluorescence intensity of the blank group) / (relative fluorescence intensity of the model group-relative fluorescence intensity of the blank group)] × 100%

[0249] The calculation results are shown in Table 2:

[0250] Table 2: Average inhibition rate of compounds on LPS-induced oxidative stress in zebrafish

[0251]

[0252]

[0253] Experimental data show that the compounds DSC194-01, DSC194-06, DSC194-11, DSC194-16, DSC194-19, DSC194-21, DSC194-25, DSC194-33, DSC194-37, DSC194-40, and DSC194-41 provided by the present invention all have obvious inhibitory effects on LPS-induced oxidative stress in zebrafish, and the inhibitory effects are stronger than those of edaravone and DSC194-00. In particular, the inhibition rates of compounds DSC194-01, DSC194-21, DSC194-40, and DSC194-41 on LPS-induced oxidative stress in zebrafish are as high as 1.5 times that of edaravone, and are at least 28% higher than the oxidative stress inhibition rate of DSC194-00, and have obvious advantages in oxidative stress inhibition activity.

[0254] Example 20: Parallel Artificial Membrane Permeation Model (PAMPA) Experiment

[0255] The compound was diluted with a pH 7.4 buffer to a 25 μg / mL solution; pig brain lipid extract (PBL) was dissolved in dodecane to prepare a 20 mg / mL solution as a phospholipid membrane; 4 μL of PBL solution was dripped onto the polyvinyl fluoride membrane of a 96-well filter plate to form a phospholipid membrane that simulates the brain environment; 300 μL / well buffer was added above the phospholipid membrane as a receptor tube, and 150 μL / well of a 25 μg / mL compound solution was added to another 96-well plate as a donor tube, with three parallel wells for each drug; the two plates were overlapped so that the phospholipid membrane could contact the donor solution to form a sandwich structure, and placed in a constant temperature environment at 37°C for 18 hours; the solution in the 96-well filter plate was taken out and transferred to a blank 96-well plate, and the OD value was measured at 340 nm. The experiment was repeated 3 times.

[0256] Permeability Pe value. The results are shown in Table 3.

[0257] Table 3: Artificial membrane permeation model (PAMPA) test results

[0258]

[0259]

[0260] The data show that the ability of the compounds DSC194-01, DSC194-06, DSC194-11, DSC194-16, DSC194-19, DSC194-21, DSC194-25, DSC194-33, DSC194-37, DSC194-40 and DSC194-41 disclosed in the present invention to penetrate the blood-brain barrier is more than 2 times higher than that of edaravone and DSC194-00. Among them, the ability of DSC194-06, DSC194-16, DSC194-21, DSC194-33, DSC194-37, DSC194-40 and DSC194-41 to penetrate the blood-brain barrier is more than 3 times that of edaravone and DSC194-00. Among them, the ability of DSC194-06, DSC194-16, DSC194-40 and DSC194-41 to penetrate the blood-brain barrier is 4 times higher than that of edaravone and DSC194-00. The compounds disclosed in the present invention show excellent ability to penetrate the blood-brain barrier.

[0261] Example 21: Safety evaluation of compounds

[0262] Normally developed 9hpf wild-type zebrafish embryos were randomly selected and placed in a 96-well plate, with 1 zebrafish embryo in each well, 20 embryos in a group, 100μL of sample solution was added to each well to maintain a concentration of 1.0μM, and an equal volume of water was added to the blank control group. Then, the embryos were placed in a 28°C incubator and incubated until 96hpf. The stock rate and deformity rate of the fish embryos were observed and counted using a stereo microscope. The results are shown in Table 4:

[0263] Table 4: Effects of compounds on zebrafish embryos

[0264]

[0265]

[0266] The data showed that at a concentration of 1.0 μM, compounds DSC194-06, DSC194-16, DSC194-21, DSC194-33, DSC194-37, DSC194-40 and DSC194-41 had zero mortality and zero deformity rates on zebrafish embryos up to 96 hpf, indicating that compounds DSC194-06, DSC194-16, DSC194-21, DSC194-33, DSC194-37, DSC194-40 and DSC194-41 have low toxicity and have the prospect of being developed into suitable dosage forms of drugs.

[0267] Example 22: Pharmacokinetic studies

[0268] 78 SD rats, half male and half female, weighing 200-250g, were divided into 13 groups, 6 rats in each group, half male and half female, respectively, control group: edaravone, DSC194-00, test group: DSC194-01, DSC194-06, DSC194-11, DSC194-16, DSC194-19, DSC194-21, DSC194-25, DSC194-33, DSC194-37, DSC194-40 and DSC194-41, all of which were administered by gavage, and the dosage was 20mg / kg. The concentration of each compound in the plasma of each group of rats was determined by high performance liquid chromatography, and the pharmacokinetic parameters were calculated by DAS2.0 program. The average values ​​of the pharmacokinetic parameters of the compounds disclosed in the present invention are shown in Table 5.

[0269] Table 5: Mean values ​​of pharmacokinetic parameters

[0270] parameter <![CDATA[AUC 0-t / mg.h.L -1 ]]> <![CDATA[MRT 0-t / h]]> <![CDATA[t max / h * ]]> <![CDATA[C max / mg.L -1* ]]> <![CDATA[T 1 / 2 / h]]> Edaravone 13.235 3.719 2.5 3.107 2.194 DSC194-00 14.516 4.011 2.3 3.115 2.206 DSC194-01 32.174 5.315 2.0 8.324 3.014 DSC194-06 31.715 4.281 2.0 8.120 2.891 DSC194-11 28.455 3.776 2.0 7.671 2.954 DSC194-16 31.019 3.891 2.0 7.415 2.012 DSC194-19 29.119 3.567 1.5 7.123 1.547 DSC194-21 35.532 4.132 2.0 7.541 2.019 DSC194-25 32.125 4.565 2.0 7.013 2.210 DSC194-33 33.234 5.342 2.0 7.001 3.171 DSC194-37 34.123 5.401 2.0 8.123 2.893 DSC194-40 39.538 6.024 2.0 8.375 3.109 DSC194-41 39.614 6.110 2.0 8.425 3.209

[0271] Note: * is the measured value

[0272] The data show that the compounds DSC194-01, DSC194-06, DSC194-11, DSC194-16, DSC194-19, DSC194-21, DSC194-25, DSC194-33, DSC194-37, DSC194-40, and DSC184-41 disclosed in the present invention have a shorter peak time (t max ) was shortened, the area under the drug-time curve (AUC) and peak concentration (C max ) were significantly increased, the mean residence time (MRT) of DSC194-40 and DSC194-41 was significantly prolonged, and the half-life (T 1 / 2 ) is extended by nearly 1.5 times.

[0273] Through the above specific description of this patent, those skilled in the art can thoroughly understand the characteristics of the present invention. At the same time, the improved results of the present invention also fall within the scope of the claims attached to this application.

Claims

1. A compound of formula I: including isomers, solvates, isotopic derivatives, oxides, and salts thereof; in: R0 is hydrogen, Y1 and Y2 are each independently CH or N; R1, R2, and R3 are each independently hydrogen, deuterium, C 1-6 Alkyl or deuterated C 1-6 alkyl; R4 is hydrogen, deuterium, C 1-6 Alkyl, C 1-6 Alkylamino, or C 1-6 Alkoxy; R5 is hydrogen, deuterium, C 1-6 Alkyl, C 1-6 Alkylamino, C 1-6 Alkyl alcohol, or Wherein, m is 1, 2 or 3; n is 1, 2 or 3; When R5 is When R0 is hydrogen, When R5 is hydrogen, deuterium, C 1-6 Alkyl, C 1-6 Alkylamino, C 1-6 In the case of alkoxy, R0 is R6 is hydrogen, deuterium, C 1-6 Alkyl, C 1-6 Alkylamino, C 1-6 Alkoxy; R7 is C 1-6 Alkylamino, C 6-12 Alkyl, C 6-12 Alkoxy; R8 and R9 are each independently hydrogen, C 1-6 Alkylamino, C 6-12 Alkyl, C 6-12 Alkoxy, and R8 and R9 are not hydrogen at the same time; R 10 C 1-6 Alkylamino, C 1-6 Alkyl carboxylic acid, or -(CR 12 R 13 ) p NR 14 CO(CR 15 R 16 ) q R 17 ; R 11 C 1-6 Alkyl, or -CO(CR 18 R 19 ) y R 20 ; Wherein, p is 2 or 3; q is 1, 2, or 3; y is 1, 2, or 3; R 12 , R 13 are independently hydrogen, C 1-6 alkyl; R 14 For hydrogen, C 1-6 alkyl; R 15 , R 16 are independently hydrogen, amino, C 1-6 alkyl; R 17 C 1-6 Alkylamino, C 6-12 Alkyl, C 6-12 Alkoxy, C 1-6 Alkyl carboxylic acids; R 18 , R 19 are independently hydrogen, amino, C 1-6 alkyl; R 20 C 1-6 Alkylamino, C 6-12 Alkyl, C 6-12 Alkoxy, C 1-6 Alkyl carboxylic acid.

2. The compound according to claim 1, having the structure of the following formula II: The substituents in formula II are as defined in formula I.

3. The compound according to claim 1 has the structure of the following formula III: The substituents in formula III are as defined in formula I.

4. The compound according to claim 1 has the structure of the following formula IV: The substituents in formula IV are as defined in formula I.

5. The compound according to claim 1, having the structure of the following formula V: The substituents in formula V are as defined in formula I.

6. The compound of claim 1, having the structure of the following formula VI: The substituents in formula VI are as defined in formula I.

7. The compound of claim 1, having the structure:

8. A pharmaceutical composition, characterized in that It comprises a therapeutically effective amount of the compound and salt thereof according to any one of claims 1 to 7 and a pharmaceutically acceptable carrier, adjuvant, vehicle and a combination thereof.

9. Use of the compound according to any one of claims 1 to 7 and its salts, solvates, isomers and polymorphs in the preparation of drugs for treating free radical-induced diseases.

10. The use according to claim 9, characterized in that It can be used to treat diseases caused by oxidative stress and / or thrombosis caused by free radicals.

11. The use according to claims 9 to 10, characterized in that The diseases caused by oxidative stress and / or thrombosis caused by free radicals include arteriosclerosis, Alzheimer's disease, Parkinson's disease, multiple sclerosis, amyotrophic lateral sclerosis, heart failure, heart disease, cerebral stroke, myocardial ischemia and ischemia-reperfusion injury, myocardial infarction, coronary heart disease, or heart failure.

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