CN118063397A - Thiobarbituric acid derivative with antioxidation effect, preparation method and application - Google Patents

Thiobarbituric acid derivative with antioxidation effect, preparation method and application Download PDF

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CN118063397A
CN118063397A CN202410022985.1A CN202410022985A CN118063397A CN 118063397 A CN118063397 A CN 118063397A CN 202410022985 A CN202410022985 A CN 202410022985A CN 118063397 A CN118063397 A CN 118063397A
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thiobarbituric acid
acid derivative
compound
preparation
dmso
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孔杜林
吴建章
沈梦雅
郭鑫
陈寅琪
陈训
钟霞
宋芸
崔晓琳
田皓宇
韦文彦
陈妍
吴文灿
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Oujiang Laboratory
Hainan Medical College
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Oujiang Laboratory
Hainan Medical College
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Abstract

The invention belongs to the field of pharmaceutical chemistry, and particularly relates to a thiobarbituric acid derivative with an antioxidant effect, a preparation method and application thereof. The thiobarbituric acid derivative provided by the invention shows effective anti-oxidation cytoprotective activity, in particular to a compound 5 with a substituent of 5- ((2-aminonaphthalene-1-yl) (4-chlorophenyl) methyl). Compound 5 also scavenges cellular ROS accumulation and reduces MDA levels by promoting Nrf2 entry into the nucleus and inhibiting expression of its downstream protein HO-1. Compound 5 can significantly reduce the cerebral infarct size and improve its brain function against CIRI lesions. In summary, the present invention reports for the first time that thiobarbituric acid derivatives with specific substituents as a novel class of potent brain protectants against CIRI injury by up-regulating Nrf2 signaling pathway in vitro and in vivo to enhance the endogenous antioxidant system.

Description

Thiobarbituric acid derivative with antioxidation effect, preparation method and application
Technical Field
The invention belongs to the field of pharmaceutical chemistry, and particularly relates to a thiobarbituric acid derivative with an antioxidant effect, a preparation method and application thereof.
Background
Cerebral stroke is one of the leading causes of death and disability worldwide. For ischemic stroke, the primary therapeutic goal is to effectively open occluded blood vessels by thrombolytic therapy or endovascular clot removal (EVT). Cerebral Ischemia Reperfusion Injury (CIRI) caused by rapid reperfusion is the primary cause of treatment inefficiency. Clearly, there is an urgent need to identify new targeted therapeutic regimens based on the pathophysiological mechanisms of CIRI.
Oxidative stress has been identified as a critical pathway leading to CIRI development, which is a trigger for neurological dysfunction and death through excessive production of peroxides and consumption of antioxidants. Antioxidant therapy has been investigated to reduce the extent of CIRI. Two types of small molecule antioxidants have been used to protect against oxidation and electrophilic toxicity. Direct antioxidants as radical scavengers require replenishment or regeneration. Edaravone is the only direct antioxidant available in japan for use in the treatment CIRI, however, the effectiveness of edaravone is not yet clear. As a radical generation inhibitor, indirect antioxidants activate the nuclear factor E2-associated factor 2 (Nrf 2) pathway, resulting in increased expression of protein products of genes involved in detoxification and clearance of active oxides and electrophiles, such as NAD (P) H quinone oxidoreductase 1 (NQO 1) and heme oxygenase 1 (HO-1), through conjugation reactions and increased antioxidant capacity of cells. To date, most of the antioxidant therapies in research are indirect antioxidants, but no approval for clinical use has been obtained.
Disclosure of Invention
The invention aims to overcome the defects and the shortcomings of the prior art and provide a thiobarbituric acid derivative with an antioxidant effect, a preparation method and application.
In a first aspect of the present invention there is provided a thiobarbituric acid derivative having a chemical structure of formula I,
Specifically, the thiobarbituric acid derivative is one of the following compounds:
according to the invention, a plurality of thiobarbituric acid derivatives with 2-benzyl-1-anilino groups are synthesized, wherein the compounds are screened out, and the thiobarbituric acid derivatives have the protection activity of resisting H 2O2 induced injury in vitro.
Among them, compounds 1-3 and compounds 5-7 have better cytoprotective effect than TBHQ (tertiary butylhydroquinone).
Of these, compound 5 having a 5- ((2-aminonaphthalen-1-yl) (4-chlorophenyl) methyl) substituent showed optimal activity.
In a second aspect of the present invention, there is provided a process for the preparation of thiobarbituric acid derivatives as described above, having the formula:
The thiobarbituric acid derivative hybrid is synthesized through a novel serial three-component reaction, and the mechanism is that aniline or 2-naphthylamine and a Knoevenagel condensation product of thiobarbituric acid and corresponding aldehyde undergo a conjugate addition reaction.
In a third aspect of the invention, the use of a thiobarbituric acid derivative as described above in the preparation of a medicament for the treatment of oxidative damage.
In a fourth aspect of the invention, the use of a thiobarbituric acid derivative as described above in the preparation of an Nrf2 activator.
In a fifth aspect of the invention, the use of a thiobarbituric acid derivative as defined above for the preparation of a medicament having the function of preventing or treating cerebral ischemia reperfusion injury diseases.
In a sixth aspect of the present invention, a pharmaceutical composition having a function of preventing or treating cerebral ischemia reperfusion injury diseases, comprising a therapeutically effective amount of an active ingredient and pharmaceutically acceptable pharmaceutical excipients; the active ingredient comprises the thiobarbituric acid derivative or the pharmaceutically acceptable salt derivative thereof.
Wherein, the "pharmaceutical excipients" refer to conventional pharmaceutical carriers in the pharmaceutical field, such as: diluents such as starch, sucrose, dextrin, lactose, pregelatinized starch, microcrystalline cellulose, calcium phosphate, and the like; wetting agents such as distilled water, ethanol; binders such as starch slurry, cellulose derivatives, povidone, gelatin, polyethylene glycol, sodium alginate solution, etc.; disintegrants such as dry starch, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, effervescent disintegrants, etc.; lubricants such as magnesium stearate, silica gel micropowder, talc, hydrogenated vegetable oil, polyethylene glycols, sodium lauryl sulfate, etc.; colorants such as titanium dioxide, sunset yellow, methylene blue, medicinal iron oxide red, and the like; other adjuvants such as flavoring agent, sweetener, etc. can also be added into the composition.
The various dosage forms of the pharmaceutical composition of the present invention used may be prepared according to conventional production methods in the pharmaceutical arts. For example by mixing the active ingredient with one or more carriers and then forming it into the desired dosage form. The preparation forms of the medicine comprise granules, injection, tablets, capsules, aerosols, suppositories, films, dripping pills, ointments, controlled release or sustained release agents or nano preparations. The present invention may be administered to a patient in need of such treatment by oral, nasal inhalation, rectal or parenteral administration in the form of a composition. For oral administration, it can be formulated into conventional solid preparations such as tablets, powders, granules, capsules, etc., and into liquid preparations such as water or oil suspensions or other liquid preparations such as syrups, elixirs, etc.; for parenteral administration, it may be formulated as a solution for injection, a water or oil suspension, or the like.
The thiobarbituric acid derivative provided by the invention shows effective cytoprotective activity in an H 2O2 induced injury model, and establishes a quantitative structure-activity relationship (QSAR) model with regression coefficient R 2 = 0.974938 through a random forest algorithm (RF). Of all these compounds, compound 5, which has a 5- ((2-aminonaphthalen-1-yl) (4-chlorophenyl) methyl) substituent, showed no significant cytotoxicity and excellent cytoprotective effect against oxidative damage. In addition, further studies were performed on compound 5 in vitro and in vivo. In keeping with its cytoprotective activity, compound 5 also scavenges cellular reactive oxygen Radical (ROS) accumulation and reduces Malondialdehyde (MDA) levels by promoting Nrf2 entry into the nucleus and inhibiting expression of its downstream protein HO-1. Administration of compound 5 significantly reduced rat cerebral infarction area and improved brain function against CIRI lesions. In summary, the present invention reports for the first time that thiobarbiturate derivatives with specific substituents are a novel class of effective brain protectants against CIRI injury by up-regulating Nrf2 signaling pathway in vitro and in vivo to enhance endogenous antioxidant system.
Drawings
In order to more clearly illustrate the embodiments of the invention or the technical solutions of the prior art, the drawings which are required in the description of the embodiments or the prior art will be briefly described, it being obvious that the drawings in the description below are only some embodiments of the invention, and that it is within the scope of the invention to one skilled in the art to obtain other drawings from these drawings without inventive faculty.
FIG. 1 shows the results of experiments on the protective activity of compounds 1-26 against H 2O2 -induced damage to PC12 cells, TBHQ was used as a positive control;
FIG. 2 is a QSAR model of PC12 cell viability pre-treated with compounds 1-26;
FIG. 3 is the protective effect of Compound 5 on H 2O2 -induced PC12 cell damage: (A) Compound 5 was resistant to H 2O2 -induced injury; (B) Compound 5 improved cell morphology under H 2O2 injury; (C) compound 5 reduces intracellular MDA content; (D) compound 5 reduces intracellular ROS accumulation;
FIG. 4 is a compound 5 activating an Nrf2 signaling pathway, (A) compound 5 promoting Nrf2 translocation; (B) is the effect of Compound 5 on HO-1 expression; (C) silencing expression of Nrf2 in PC12 cells using siRNA; (D) Effect of silencing of Nrf2 on antioxidant activity of compound 5;
FIG. 5 shows the protective effect of Compound 5 on MCAO-induced CIRI, (A) is a representative sample of TTC-stained brain tissue sections; (B) infarct size and nerve score.
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings, for the purpose of making the objects, technical solutions and advantages of the present invention more apparent.
Example 1 synthesis of thiobarbituric acid derivatives:
Compounds 1-26 were synthesized by a novel series three-component reaction in which aniline or 2-naphthylamine was subjected to a conjugate addition reaction with the Knoevenagel condensation product of thiobarbituric acid and the corresponding aldehyde.
All reactions were carried out in a oven-dried glassware equipped with magnetic stirring. Unless otherwise indicated, all reagents were purchased from commercial suppliers and used without further purification. All solvents were purified and dried according to standard methods prior to use. The reaction was monitored by Thin Layer Chromatography (TLC) on silica gel pre-coated glass plates. The plate was visualized using 254nm uv radiation.
To a mixture of Ethyl Lactate (EL). Times.water (3:2, 4 mL) was added aromatic amine (1 mmol), 2-thiobarbituric acid (1 mmol) and aldehyde (1 mmol). The mixture was stirred at room temperature or 50 ℃ for 10 hours and monitored by TLC. After completion, the reaction mixture was filtered and the crude powder was purified by recrystallization from ethanol. The spectral data for the novel or unreported compounds are as follows:
5- ((2-aminonaphthalen-1-yl) (phenyl) methyl) -2-thiodihydropyrimidine-4, 6 (1 h,5 h) -dione (1): white solid, melting point: 229-230 ℃, yield 90％.1H NMR(400MHz,DMSO-d6):δ＝11.59(m,1H),10.83(s,1H),9.42(s,1H),9.27(s,1H),7.81-7.90(m,3H),7.42(d,J＝8Hz,1H),7.33-7.36(m,1H),7.17-7.30(m,6H),5.26(s,1H),4.07(m,1H)ppm.13C NMR(100MHz,DMSO-d6):δ＝181.3,169.2,165.1,140.9,135.5,131.0,130.2,128.9,128.7,128.7,127.5,127.2,124.1,122.6,117.0,115.5,55.5,41.1ppm.HRMS(ESI)[M-H]-Calcd for C21H16N3O2S:374.0969,found 374.0962.
5- ((2-Aminonaphthalen-1-yl) (p-tolyl) methyl) -2-thiodihydropyrimidine-4, 6 (1 h,5 h) -dione (2), white solid, melting point: 241-242 ℃, yield 92％.1H NMR(400MHz,DMSO-d6):δ＝11.61(s,1H),10.82(s,1H),9.43(s,1H),9.28(s,1H),7.84-7.87(m,2H),7.80(d,J＝8Hz,1H),7.41(d,J＝8Hz,1H),7.27-7.36(m,2H),7.02-7.12(m,4H),5.21(s,1H),4.05(s,1H),2.20(s,3H)ppm.13C NMR(100MHz,DMSO-d6):δ＝181.4,169.3,165.1,137.8,136.4,135.5,131.0,130.1,129.2,128.8,128.6,127.9,127.4,127.1,124.0,122.5,117.0,115.7,55.7,20.6ppm.HRMS(ESI)[M-H]-Calcd for C22H18N3O2S:388.1125,found 388.1124.
5- ((2-Aminonaphthalen-1-yl) (4-methoxyphenyl) methyl) -2-thiodihydropyrimidine-4, 6 (1 h,5 h) -dione (3): white solid, melting point: 211-212 ℃, yield 91％.1H NMR(400MHz,DMSO-d6):δ＝11.58(s,1H),10.79(s,1H),9.39(s,1H),9.25-9.26(m,1H),7.85-7.87(m,2H),7.79(d,J＝8Hz,1H),7.43(t,J＝8Hz,1H),7.35(t,J＝8Hz,1H),7.26-7.28(m,1H),7.11-7.13(m,2H),6.80-6.83(m,2H),5.18(s,1H),4.02(s,1H),3.67(s,3H)ppm.13C NMR(100MHz,DMSO-d6):δ＝181.3,169.3,165.1,158.3,135.4,132.6,131.0,130.1,128.8,128.6,127.1,124.0,122.5,117.0,115.9,114.1,55.8,55.0,40.4ppm.HRMS(ESI)[M-H]-Calcd for C22H18N3O3S:404.1074,found 404.1076.
5- ((2-Aminonaphthalen-1-yl) (4- (tert-butyl) phenyl) methyl) -2-thiodihydropyrimidine-4, 6 (1 h,5 h) -dione (4): white solid, melting point: 227.5-228.5 ℃, yield 82％.1H NMR(400MHz,DMSO-d6):δ＝11.56(s,1H),10.81(s,1H),9.39(s,1H),9.26(s,1H),7.80-7.87(m,3H),7.41-7.44(m,1H),7.34(d,J＝8Hz,1H),7.21-7.28(m,3H),7.13-7.15(m,2H),5.20(s,1H),4.04(s,1H),1.19(s,9H)ppm.13C NMR(100MHz,DMSO-d6):δ＝181.4,169.3,165.2,149.5,137.8,135.4,131.0,130.2,128.8,128.7,127.1,125.5,124.1,122.6,117.0,115.8,55.6,40.7,34.2,31.1ppm.HRMS(ESI)[M-H]-Calcd for C25H24N3O2S:430.1595,found 430.1598.
5- ((2-Aminonaphthalen-1-yl) (4-chlorophenyl) methyl) -2-thiodihydropyrimidine-4, 6 (1 h,5 h) -dione (5): white solid, melting point: 239-240 deg.c, yield 84％.1H NMR(400MHz,DMSO-d6):δ＝11.60(s,1H),10.85(s,1H),9.40(s,1H),9.24(s,1H),7.85-7.89(m,2H),7.78-7.80(m,1H),7.41-7.45(m,1H),7.33-7.38(m,3H),7.27-7.29(m,1H),7.22-7.24(m,2H),5.28(s,1H),4.03(s,1H)ppm.13C NMR(100MHz,DMSO-d6):δ＝181.3,169.0,164.9,139.8,135.6,131.9,130.2,129.4,129.1,128.7,127.3,124.2,122.4,117.0,115.0,55.4,40.3ppm.HRMS(ESI)[M-H]-:Calcd for C21H15ClN3O2S:408.0579,found 408.0574.
5- ((2-Aminonaphthalen-1-yl) (4-bromophenyl) methyl) -2-thiodihydropyrimidine-4, 6 (1 h,5 h) -dione (6): white solid, melting point: 226-227 deg.C, yield 88％.1H NMR(400MHz,DMSO-d6):δ＝11.61(s,1H),10.86(s,1H),9.41(s,1H),9.25(s,1H),7.85-7.89(m,2H),7.78-7.80(m,1H),7.46-7.48(m,2H),7.41-7.43(m,1H),7.34-7.37(m,1H),7.28-7.30(m,1H),7.16-7.18(m,2H),5.27(s,1H),4.04(s,1H)ppm.13C NMR(100MHz,DMSO-d6):δ＝181.3,169.0,164.9,140.2,135.6,131.6,130.9,130.2,129.8,128.7,127.3,124.2,122.4,117.0,114.9,55.3,40.4ppm.HRMS(ESI)[M-H]-:Calcd for C21H15BrN3O2S:452.0074,found 452.0065.
5- ((2-Aminonaphthalen-1-yl) (4- (trifluoromethyl) phenyl) methyl) -2-thiodihydropyrimidine-4, 6 (1 h,5 h) -dione (7): white solid, melting point: 241-242 ℃, yield 92％.1H NMR(400MHz,DMSO-d6):δ＝11.64-11.63(m,1H),10.90-10.92(m,1H),9.44(s,1H),9.25(s,1H),7.86-7.91(m,2H),7.81(d,J＝8Hz,1H),7.66(d,J＝8Hz,2H),7.29-7.49(m,5H),5.41(s,1H),4.07-4.09(m,1H)ppm.13C NMR(100MHz,DMSO-d6):δ＝181.3,168.9,164.8,145.5,135.7,130.9,130.2,129.3,128.7,128.4,127.4,125.7,124.2,122.8,122.4,117.0,114.6,55.1,40.7ppm.HRMS(ESI)[M-H]-:Calcd for C22H15F3N3O2S:442.0843,found 442.0850.
5- ((2-Aminonaphthalen-1-yl) (4-nitrophenyl) methyl) -2-thiodihydropyrimidine-4, 6 (1 h,5 h) -dione (8): white solid, melting point: 234-235 deg.C, yield 93％.％.1H NMR(400MHz,DMSO-d6)δ＝11.65(s,1H),10.94(s,1H),9.45(s,1H),9.25(s,1H),8.15-8.17(m,2H),7.87-7.93(m,2H),7.81(d,J＝8Hz,1H),7.30-7.51(m,5H),5.48(s,1H),4.09(s,1H)ppm.13C NMR(100MHz,DMSO-d6)δ＝181.2,168.7,164.6,148.5,146.7,135.7,130.9,130.2,129.4,129.0,128.7,124.2,123.9,122.3,117.0,114.2,54.8,40.6ppm.HRMS(ESI)[M-H]-:Calcd for C21H15N4O4S:419.0819,found 419.0825.
5- ((2-Aminonaphthalen-1-yl) (4- (methylsulfonyl) phenyl) methyl) -2-thiodihydropyrimidine-4, 6 (1 h,5 h) -dione (9): white solid, melting point: 238-239 ℃, yield 91％.1H NMR(400MHz,DMSO-d6):δ＝11.64(s,1H),10.92(s,1H),9.43(s,1H),9.26(s,1H),7.89-7.93(m,1H),7.86-7.87(m,1H),7.81-7.84(m,2H),7.49-7.51(m,2H),7.43-7.47(m,1H),7.35-7.39(m,1H),7.33-7.31(m,1H),5.44(s,1H),4.09(s,1H),3.17(s,3H)ppm.13C NMR(100MHz,DMSO-d6):δ＝181.3,168.8,164.8,161.5,146.7,139.8,135.7,130.9,130.2,129.4,128.6,127.5,127.4,124.3,122.4,117.1,114.5,55.1,43.4,40.7ppm.HRMS(ESI)[M-H]-:Calcd for C22H18N3O4S2:452.0744,found 452.0714.
5-((2-aminonaphthalen-1-yl)(3-nitrophenyl)methyl)-2-thioxodihydropyrimidine-4,6(1H,5H)-dione(10):white solid,m.p:230-231℃,yield 90％.1H NMR(400MHz,DMSO-d6):δ＝11.67(s,1H),10.96(s,1H),9.43(s,1H),9.26(s,1H),8.29(s,1H),8.07-8.09(m,1H),7.92-7.94(m,1H),7.83-7.89(m,2H),7.50-7.57(m,2H),7.42-7.46(m,1H),7.32-7.38(m,1H),5.52(s,1H),4.11(s,1H)ppm.13C NMR(100MHz,DMSO-d6):δ＝181.3,168.8,164.7,148.2,142.9,135.9,134.1,130.4,130.2,129.6,128.8,127.5,124.4,122.7,122.5,122.4,117.1,114.3,55.2,40.4ppm.HRMS(ESI)[M-H]-:Calcd for C21H15N4O4S:419.0819,found 419.0825.
5- ((2-Aminonaphthalen-1-yl) (2-fluorophenyl) methyl) -2-thiodihydropyrimidine-4, 6 (1 h,5 h) -dione (11): white solid, melting point: 228-229 ℃, yield 86％.1H NMR(400MHz,DMSO-d6):δ＝11.61(s,1H),10.85(s,1H),9.42(s,1H),9.25(s,1H),7.87(t,J＝8Hz,2H),7.80(d,J＝8Hz,1H),7.43(d,J＝8Hz,1H),7.34-7.37(m,1H),7.23-7.29(m,3H),7.10(d,J＝8Hz,2H),5.28(s,1H),4.04(s,1H)ppm.13C NMR(100MHz,DMSO-d6):δ＝181.3,169.1,164.9,136.9,135.5,130.9,130.1,129.5,128.7,127.2,124.1,122.5,117.0,115.6,115.3,55.6,40.1,39.9,39.7,39.5,39.3,39.1,38.9ppm.HRMS(ESI)[M-H]-：Calcd for C21H15FN3O2S：392.0874,found 392.0878.
5- ((2-Aminonaphthalen-1-yl) (naphthalen-1-yl) methyl) -2-thiodihydropyrimidine-4, 6 (1 h,5 h) -dione (12): white solid, melting point: 229-230 ℃, yield 86％.1H NMR(400MHz,DMSO-d6)：δ＝11.06(s,1H),10.94(s,1H),9.49(s,1H),9.37(s,1H),8.50-8.52(m,1H),8.01-8.03(m,1H),7.89-7.93(m,1H),7.84-7.87(m,1H),7.77-7.82(m,2H),7.66(t,J＝8Hz,1H),7.50-7.53(m,1H),7.22-7.34(m,4H),6.71(d,J＝8Hz,1H),6.04(s,1H),4.12(s,1H)ppm.13C NMR(100MHz,DMSO-d6)δ＝181.1,168.5,164.6,136.5,135.5,134.1,130.9,129.2,128.9,128.7,128.0,127.2,126.2,125.6,124.6,124.1,123.4,122.5,116.9,116.5,54.5,35.9ppm.HRMS(ESI)[M-H]-：Calcd for C25H18N3O2S：424.0025,found 424.1124.
5- ((6-Amino-2, 3, 4-trimethoxyphenyl) (phenyl) methyl) -2-thiodihydropyrimidine-4, 6 (1H, 5H) -dione (13): white solid, melting point: 218-219 ℃, yield 87％.1H NMR(400MHz,DMSO-d6)：δ＝11.58(s,1H),10.45(s,1H),9.45(s,1H),9.32(s,1H),7.29(t,J＝8Hz,2H),7.20(t,J＝8Hz,3H),6.47(s,1H),4.66(s,1H),3.90(s,1H),3.78(s,3H),3.68(s,3H),3.54(s,3H)ppm.13C NMR(100MHz,DMSO-d6)：δ＝181.4,169.4,165.1,153.0,150.7,141.3,137.1,133.9,128.5,127.5,127.2,127.0,108.6,96.0,60.7,60.5,55.7,55.3ppm.HRMS(ESI)[M-H]-：Calcd for C20H20N3O5S：414.1129,found 414.1124.
5- ((6-Amino-2, 3, 4-trimethoxyphenyl) (p-tolyl) methyl) -2-thiodihydropyrimidine-4, 6 (1 h,5 h) -dione (14): white solid, melting point: 209-210 ℃ and yield 84％.％.1H NMR(400MHz,DMSO-d6)：δ＝11.59(s,1H),10.45(s,1H),9.45-9.32(m,2H),6.96-7.10(m,4H),6.46(s,1H),4.61(s,1H),,3.85(s,1H),3.77(s,3H),3.68(s,3H),3.53(s,3H),2.23(m,3H)ppm.13C NMR(100MHz,DMSO-d6)：δ＝181.4,169.4,165.1,152.9,150.7,138.2,137.1,136.1,133.9,129.0,127.1,108.7,96.0,60.7,60.5,55.7,55.5,20.6ppm.HRMS(ESI)[M-H]-：Calcd for C21H22N4O7S：428.1286,found 428.1287.
5- ((6-Amino-2, 3, 4-trimethoxyphenyl) (4-chlorophenyl) methyl) -2-thiodihydropyrimidine-4, 6 (1H, 5H) -dione (15): white solid, melting point: 222-223 deg.C, yield 87％.1H NMR(400MHz,DMSO-d6)：δ＝11.63(s,1H),10.50(s,1H),9.32-9.47(m,2H),7.36-7.38(m,2H),7.20-7.22(m,2H),6.48-6.49(m,1H),4.67(s,1H),3.84-3.91(m,1H),3.76-3.78(m,3H),3.66-3.68(m,3H),3.55(s,3H)ppm.13C NMR(100MHz,DMSO-d6)：δ＝181.4,169.2,164.9,153.2,150.8,140.2,137.2,133.9,131.8,129.2,128.6,108.0,96.1,60.8,60.6,55.7,55.3ppm.HRMS(ESI)[M-H]-：Calcd for C20H19ClN3O5S：448.0739,found 448.0741.
5- ((6-Amino-2, 3, 4-trimethoxyphenyl) (4-nitrophenyl) methyl) -2-thiodihydropyrimidine-4, 6 (1H, 5H) -dione (16): white solid, melting point: 216-217 ℃, yield 83％.1H NMR(400MHz,DMSO-d6)：δ＝11.66(s,1H),10.59(s,1H),9.48(s,1H),9.32(s,1H),8.19(d,J＝8Hz,2H),7.47-7.49(m,2H),6.50(s,1H),4.83(s,1H),3.93(s,1H),3.79(s,3H),3.67(s,3H),3.57(s,3H)ppm.13C NMR(100MHz,DMSO-d6)：δ＝181.4,168.9,164.7,153.6,150.8,149.1,146.7,137.3,134.0,128.7,123.9,107.4,96.2,60.9,60.6,55.8,54.8ppm.HRMS(ESI)[M-H]-：Calcd for C20H19N4O7S：459.0980,found 459.0972.
5- ((6-Amino-2, 3, 4-trimethoxyphenyl) (3-nitrophenyl) methyl) -2-thiodihydropyrimidine-4, 6 (1H, 5H) -dione (17): white solid, melting point: 228-229 ℃, yield 90％.1H NMR(400MHz,DMSO-d6)：δ＝11.70(s,1H),10.60(s,1H),9.49(s,1H),9.31(s,1H),8.24(s,1H),8.11(d,J＝8Hz,1H),7.49-7.63(m,2H),6.51(s,1H),4.85(s,1H),3.92(s,1H),3.80(s,3H),3.68(s,3H),3.58(s,3H)ppm.13C NMR(100MHz,DMSO-d6)：δ＝181.3,168.9,164.7,153.5,150.8,148.1,143.3,137.2,134.0,133.8,130.3,122.3,107.2,96.1,60.9,60.6,55.8,55.1ppm.HRMS(ESI)[M-H]-：Calcd for C20H19N4O7S：459.0980,found 459.0985.
5- ((2-Amino-4, 6-dimethoxyphenyl) (phenyl) methyl) -2-thiodihydropyrimidine-4, 6 (1H, 5H) -dione (18): white solid, melting point: 223-224 ℃, yield 85％.1H NMR(400MHz,DMSO-d6)：δ＝11.64(s,1H),10.50(s,1H),9.34-9.47(m,2H),7.27(t,J＝8Hz,3H),7.19(t,J＝8Hz,3H),6.22-6.25(m,2H),4.62(s,1H),3.88(s,1H),3.73-7.74(m,3H),3.67(s,3H)ppm.13C NMR(100MHz,DMSO-d6)：δ＝181.4,169.5,165.4,160.1,157.5,141.3,139.3,128.4,127.1,126.9,103.1,93.7,93.0,55.7,55.4,55.2ppm.HRMS(ESI)[M-H]-：Calcd for C19H18N3O4S：384.1024,found 384.1023.
5- ((2-Amino-4, 6-dimethoxyphenyl) (p-tolyl) methyl) -2-thiodihydropyrimidine-4, 6 (1H, 5H) -dione (19): white solid, melting point: 227-228 ℃, yield 82％.1H NMR(400MHz,DMSO-d6)：δ＝11.65(s,1H),10.48(s,1H),9.35-9.46(m,2H),6.98-7.07(m,4H),6.22-6.24(m,2H),4.59(s,1H),3.86(s,1H),3.72-7.74(m,3H),3.66(s,3H),2.22(s,3H)ppm.13C NMR(100MHz,DMSO-d6)：δ＝181.5,169.5,165.5,160.1,157.5,139.3,138.4,136.0,129.0,127.3,127.1,103.3,93.7,93.0,55.6,55.2,20.6ppm.HRMS(ESI)[M-H]-：Calcd for C20H20N3O4S：398.1180,found 398.1173.
5- ((2-Amino-5-methylphenyl) (4-chlorophenyl) methyl) -2-thiodihydropyrimidine-4, 6 (1H, 5H) -dione (20): white solid, melting point: 232-233 ℃, yield 84％.1H NMR(400MHz,DMSO-d6)：δ＝11.67(s,1H),10.54(s,1H),9.47(s,1H),9.32-9.32(m,1H),7.34-7.36(m,2H),7.20(d,J＝12Hz,2H),6.26-6.22(m,2H),4.62(s,1H),3.84(s,1H),3.73-3.74(m,3H),3.67(s,3H)ppm.13C NMR(100MHz,DMSO-d6)：δ＝181.1,169.2,165.2,160.3,157.5,140.2,139.3,131.6,129.1,128.5,102.6,93.7,93.1,55.7,55.3,55.2,33.8ppm.HRMS(ESI)[M-H]-：Calcd for C19H17ClN3O4S：418.0634,found 418.0631.
5- ((2-Amino-4, 6-dimethoxyphenyl) (4-nitrophenyl) methyl) -2-thiodihydropyrimidine-4, 6 (1H, 5H) -dione (21): white solid, melting point: 234-235 ℃, yield 81％.1H NMR(400MHz,DMSO-d6)：δ＝11.70(s,1H),10.63(s,1H),9.32-9.50(m,2H),8.16-8.19(m,2H),7.46-7.48(m,2H),6.25-6.28(m,2H),4.77(s,1H),3.90(s,1H),3.72-3.75(m,3H),3.68(s,3H)ppm.13C NMR(100MHz,DMSO-d6)：δ＝181.4,168.9,164.9,160.6,157.6,149.1,146.6,139.3,128.6,123.8,101.9,93.9,93.2,55.8,55.3,54.8,33.8ppm.HRMS(ESI)[M-H]-：Calcd for C19H17N4O6S：429.0874,found 429.0870.
5- ((2-Amino-5-methylphenyl) (phenyl) methyl) -2-thiodihydropyrimidine-4, 6 (1 h,5 h) -dione (22): white solid, melting point: 218-219 ℃, yield 82％.1H NMR(400MHz,DMSO-d6)：δ＝11.26(s,1H),10.64(s,1H),9.40(s,2H),7.38-7.41(m,2H),7.25-7.33(m,3H),6.99-7.01(m,1H),6.85(d,J＝8Hz,1H),6.33(s,1H),4.52-4.55(m,1H),4.25-4.28(m,1H),2.08(s,3H)ppm.13C NMR(100MHz,DMSO-d6)：δ＝181.1,169.8,169.2,139.4,134.5,131.6,128.8,128.6,128.3,128.1,127.5,125.6,115.3,54.0,44.2,20.5ppm.HRMS(ESI)[M-H]-：Calcd for C18H16N3O2S：338.0969,found 338.0972.
5- ((2-Amino-5-methylphenyl) (p-tolyl) methyl) -2-thiodihydropyrimidine-4, 6 (1 h,5 h) -dione (23): white solid, melting point: 234-235 deg.C, yield 80％.1H NMR(400MHz,DMSO-d6)：δ＝11.23(s,1H),10.61(s,1H),9.34-9.45(m,2H),7.12-7.20(m,4H),6.97-6.99(m,1H),6.83(d,J＝8Hz,1H),6.29(s,1H),4.48(d,J＝12Hz,1H),4.24(d,J＝12Hz,1H),2.31(s,3H),2.07(s,3H)ppm.13C NMR(100MHz,DMSO-d6)：δ＝181.1,169.9,166.3,136.6,136.2,134.5,131.5,129.4,128.5,128.2,128.0,125.9,115.3,54.0,43.7,20.7,20.5ppm.HRMS(ESI)[M-H]-：Calcd for C19H18N3O2S：352.1125,found 352.1129.
5- ((2-Amino-5-methylphenyl) (4-chlorophenyl) methyl) -2-thiodihydropyrimidine-4, 6 (1H, 5H) -dione (24): white solid, melting point: 233-234 deg.C, yield 82％.1H NMR(400MHz,DMSO-d6)：δ＝11.27(s,1,H),10.65(s,1H),9.38-9.42(m,2H),7.45-7.47(m,2H),7.27(d,J＝8Hz,2H),7.00-7.02(m,1H),6.85(d,J＝8Hz,1H),6.34(s,1H),4.57(d,J＝12Hz,1H),4.21(d,J＝12Hz,1H),2.09(s,3H)ppm.13C NMR(100MHz,DMSO-d6)：δ＝181.0,169.6,169.0,138.4,134.6,132.1,131.8,130.5,128.8,128.5,128.0,125.1,115.4,54.0,43.5,20.5ppm.HRMS(ESI)[M-H]-：Calcd for C18H15ClN3O2S：372.0579,found 372.0572.
5- ((2-Amino-5-methylphenyl) (3-nitrophenyl) methyl) -2-thiodihydropyrimidine-4, 6 (1 h,5 h) -dione (25): white solid, melting point: 221-222 ℃, yield 84％.1H NMR(400MHz,DMSO-d6)：δ＝11.36(s,1H),10.72(s,1H),9.35-9.42(m,2H),8.16-8.19(m,2H),7.68-7.70(m,2H),7.02-7.04(m,1H),6.89(d,J＝8Hz,1H),6.43(s,1H),4.80(d,J＝12Hz,1H),4.26(d,J＝8Hz,1H),2.09(s,3H)ppm.13C NMR(100MHz,DMSO-d6)：δ＝181.0,169.3,165.6,148.1,142.0,135.3,134.7,132.0,130.4,128.8,128.2,124.2,123.4,122.7,115.6,54.1,43.8,20.4ppm.HRMS(ESI)[M-H]-：Calcd for C18H15N4O4S：338.0819,found 338.0818.
5- ((2-Amino-5-ethylphenyl) (4-nitrophenyl) methyl) -2-thiodihydropyrimidine-4, 6 (1H, 5H) -dione (26): white solid, melting point: 223.5-224.5 ℃ and yield 88％.1H NMR(400MHz,DMSO-d6)：δ＝11.33(s,1H),10.73(s,1H),9.36-9.43(m,2H),8.27(d,J＝8Hz,2H),7.55(d,J＝8Hz,2H),7.06-7.08(m,1H),6.91(d,J＝8Hz,1H),6.38(s,1H),4.78(d,J＝8Hz,1H),4.27(d,J＝12Hz,1H),2.35-2.41(m,2H),1.00(t,J＝8Hz,3H)ppm.13C NMR(100MHz,DMSO-d6)：δ＝181.0,169.3,165.6,147.5,146.9,138.4,134.9,130.0,127.5,127.0,124.2,124.0,115.6,53.9,43.9,27.5,15.6ppm.HRMS(ESI)[M-H]-：Calcd for C19H17N4O4S：397.0976,found 397.0973.
Example 2 protective Activity of PC12 cells against H 2O2 induced injury in vitro:
1. Cell culture
The rat pheochromocytoma cell line (PC 12 cells) was purchased from the institute of biochemistry and cell biology, academy of sciences, china Shanghai. Cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% fetal bovine serum (FBS, gibco) of 1% penicillin-streptomycin, maintained at 37 ℃ in a humidified incubator with 95% air and 5% co 2 and serum starved by synchronization prior to stimulation with hydrogen peroxide (H2O 2) or test compounds.
2. Test for protective Activity of Compounds against H 2O2 -induced oxidative damage of PC12 cells and test for cytotoxicity of Compounds
The 3- [4,5-Dimethylthiazol-2-yl ] -2,5-diphenyltetrazolium bromide, thiazolyl blue (MTT) MTT assay was used to detect toxicity of test compounds and viability of PC12 cells. The compound was dissolved in dimethyl sulfoxide (DMSO, sigma Aldrich, shanghai, china) and diluted with medium. In the cytoprotection experiments, cells were incubated with different concentrations of test compound for 18 hours, followed by induction of cell damage with 500 μMH 2O2 for 24 hours. MTT solution (20. Mu.L, 5 mg/mL) was added to PC12 cells, followed by further incubation at 37℃and 5% CO 2 for 4 hours. The positive control was TBHQ (SIGMA ALDRICH, shanghai, china). The MTT solution was then discarded and DMSO (120 μl) was added to dissolve the formazan crystals. Absorbance was measured at 490 nm wavelength using a BioTek microplate detector to determine cell viability. In the cytotoxicity test, cells were incubated with fresh medium containing different concentrations (2.5, 5, 10 μm) of test compound for 42 hours.
As shown in FIG. 1, of 26 compounds, compounds 1-3, 5-12, 14-15, 17-18, 20-22 and 24 exhibited some protective activity against H 2O2 -induced injury. It was found that compound 5, which had a 5- ((2-aminonaphthalen-1-yl) (4-chlorophenyl) methyl) substituent, showed the highest activity. Of these compounds with 2-aminonaphthalen-1-yl, the compound with electron-rich groups (compounds 2 and 3) or halogen (compounds 5 and 6) at the para position of the benzene ring of compound 1 shows similar or even stronger better cytoprotective properties as TBHQ (tertiary butylhydroquinone) at the same concentration (5 μm). TBHQ has been widely demonstrated to have cytoprotective effects against CIRI lesions. However, as an exception, compound 4, which carries a tert-butyl group in the para position of the phenyl group, shows a lower cytoprotective effect. In general, compounds having electron withdrawing groups on the benzene ring (compounds 8,9, 10 and 11) exhibit lower protective activity. Except that compound 7 having a strong electron withdrawing CF 3 group at the para-position of the benzene ring showed higher protection (79% cell viability) than TBHQ, whereas compound 9 having two naphthalene groups showed slightly lower activity than TBHQ. In general, when the C-5 diarylmethyl substituent of the thiobarbituric acid derivative carries the same substituted phenyl group, the substituted phenyl group as the second aryl group has a lower cytoprotective effect than TBHQ (Compounds 13-26).
To further investigate the exact protective activity, PC12 cells were pretreated with different concentrations of compound 5. As shown in fig. 3A, cell viability exhibited dose dependency. Compound 5 as low as 2.5 μm increased cell viability compared to H 2O2 group. As shown in fig. 3B, H 2O2 treatment had greater morphological damage to PC12 cells than the control group. Pretreatment with compound 5 (2.5. Mu.M, 5. Mu.M, and 10. Mu.M) restored abnormalities in PC12 cells caused by H 2O2. More importantly, 10 μm of compound 5 significantly restored cell morphology compared to the H 2O2 group.
3. Quantitative structure-effect relation model
Quantitative structure-activity relationships (QSARs) are frequently used in new drug development due to their guiding effect on drug design. The various QSAR models built by machine learning algorithms perform well. The QSAR model may be constructed by a random forest algorithm, which is a classifier that contains a number of decision trees over various subsets of a given dataset and averages to improve the predictive accuracy of the dataset. And establishing a QSAR model by utilizing sci-learn machine learning library and Python language. Molecular descriptors of molecular properties were calculated using RDKit, and the results included 124-dimensional molecular descriptors, topological polar surface area, partition coefficients, molecular weights, and the like. These descriptors are input into a random forest containing 800 default subtrees, and a QSAR model is built through computer training.
According to the activity of PC12 cells in the cell experiment, a QSAR model is established by adopting a random forest algorithm. As shown in fig. 2, the values on the horizontal axis show experimental cell viability and the values on the vertical axis show the calculated values of the QSAR model. Regression coefficient value is 0.974938, which shows that the fitting effect is good.
MDA assay
Malondialdehyde (MDA) is one of the end products of lipid peroxidation in cells. MDA levels are generally considered to be a marker of oxidative stress.
PC12 cells were seeded at a density of 3×10 5 cells/well in 6-well plates for 24 hours, then pretreated with compound 5 for 18 hours, followed by 1.25mM H 2O2 for 3 hours to induce cell damage. Protein samples were collected and quantified using the lipid peroxidation MDA detection kit (institute of bioengineering, south kyo, china).
As shown in fig. 3C, H 2O2 treatment significantly increased MDA accumulation in PC12 cells. Pretreatment with compound 5 (2.5. Mu.M, 5. Mu.M, and 10. Mu.M) reduced MDA levels in PC12 cells, indicating antioxidant activity of compound 5.
5. Reactive oxygen species detection
To confirm the antioxidant effect of compound 5 in PC12 cells, it was further examined whether compound 5 could reduce intracellular ROS accumulation using the DCFH-DA method.
Intracellular ROS content was assessed using dichlorofluorescein diacetate (DCFH-DA, biyun Biotechnology Co., shanghai, china). Cells were seeded into 6-well plates at a concentration of 3×10 5 cells and cultured for 24 hours. After 18 hours of pre-incubation with the test compound, the cells were exposed to 1.25mM H2O2 for 3 hours, followed by staining in starvation medium containing 1. Mu.L DCFH-DA per ml at 37℃for 30 minutes in the absence of light. After washing off the excess probe with PBS, the fluorescence intensity was observed by a fluorescence microscope (Nikon ECLIPSE TIES, nikon Ltd, japan).
As shown in fig. 3D, the H 2O2 group detected significantly stronger green fluorescence compared to the DMSO group, while the green fluorescence was significantly weaker in PC12 cells pretreated with compound 5. Furthermore, the decrease in green fluorescence intensity depends on the increase in the concentration of compound 5. These findings indicate that reducing intracellular ROS accumulation may play a key role in the beneficial effect of compound 5 on anti-H 2O2 -induced PC12 cell damage.
6. Immunofluorescence
To further elucidate the antioxidant mechanism of compound 5, nrf2 was localized in PC12 cells by immunofluorescence.
PC12 cells were incubated in 12 well plates at a density of 8 x 10 4 cells for 24 hours. After 6 hours of pre-incubation with the test compound, cells were fixed with 4% paraformaldehyde for 20 minutes at room temperature and washed 3 times with PBS. Cells were permeabilized in 0.1% Triton X-100 (Sigma-Aldrich) for 15 min and blocked in 2% BSA for 1 hr at room temperature. After washing, cells were incubated with 1:200 dilution of Nrf2 primary antibody (sc-13032,Santa Cruz Biotechnology,USA,1:200) overnight at 4 ℃ and anti-mouse IgG-PE (sc-13032, goat) with fluorescent mouse antibody, santa Cruz Biotechnology, U.S. 1:200), at room temperature for 1 hour. Washed 3 times with PBS and then incubated with DAPI (Beyotime Biotech, china) for 8 minutes in the dark. After sealing with a neutral resin, photographs were taken under a fluorescence microscope (nikon, japan).
Red fluorescence represents Nrf2, blue fluorescence pen nuclei. As shown in fig. 4A, blue fluorescence in the DMSO group was surrounded by red fluorescence, indicating that Nrf2 was mainly present in the cell cytoplasm. In contrast, more overlapping red and blue fluorescence was observed in PC12 cells pretreated with compound 5, indicating increased expression of Nrf2 in the nucleus.
Western blot detection of nrf2
PC12 cells were seeded in 6-well plates at a density of 3X 10 5 and incubated at 37℃for 24-48 hours. After appropriate treatment, cells were collected and lysed with ice-cold RIPA lysis buffer. An equal amount of protein sample (80. Mu.g) was separated on a 10% SDS-PAGE gel and transferred to nitrocellulose membrane by electrophoresis. Subsequently, the membranes were blocked with 5% skim milk and incubated with primary antibodies overnight at 4 ℃): anti-HO-1 (sc-10789,Santa Cruz Biotechnology,1:300), anti-GADPH (sc-47724, santa Cruz Biotechnology, 1:1000). After 3 washes with 1 XTBST, the membranes were incubated with anti-rabbit IgG for 1h at room temperature. Proteins were visualized by exposure to a Chemidoc XRS+ system (Bio-Rad, hercules, calif., USA). Band density was quantified by Image J software (national institutes of health, bescens, maryland).
Increased expression of antioxidant enzyme HO-1 was detected in PC12 cells pretreated with Compound 5 (FIG. 4B).
8. Transfection assay
Small interfering RNAs (siRNAs) targeting Nrf2 are provided by Shanghai Ji Ma (siRNA sequence, sense: 5'-GGUUCAGUGACUCGGAAAUTT-3', antisense: 5 '-AUUUCCGAGUCACU-GAACCTT-3'). For the transfection procedure, PC12 cells were seeded at a density of 3×10 5 in 6-well plates for 24 hours, washed with PBS, and Nrf2 siRNA transfected using Lipofectamine TM 2000 transfection reagent (Invitrogen, carlsbad, CA, USA). During transfection, cells are incubated with starvation medium to increase transfection efficiency. After 10 hours of transfection, the original starvation medium was replaced with medium containing fetal bovine serum and the transfected cells were cultured for further experiments.
Expression of Nrf2 in PC12 cells could be silenced using siRNA (fig. 4C). As shown in fig. 4D, the viability of Nrf 2-silenced PC12 cells pretreated with compound 5 was significantly reduced compared to the protective effect of compound 5 on H 2O2 -induced damage to normal PC12 cells, indicating that the antioxidant activity of compound 5 was associated with the Nrf2 signaling pathway.
The above results indicate that compound 5 exerts cytoprotective effects on oxidative damage of cells by activating Nrf2 and HO-1 signaling pathways.
Example 3 antioxidant Activity in vivo:
the MCAO/R model was used to evaluate the antioxidant activity of compound 5 in vivo.
C57BL/6 mice, 23-25g in weight, were purchased from Henan province laboratory animal center (Zhengzhou, china). All mice were placed in polypropylene cages with a light and dark cycle at 25.+ -. 2 ℃ and a relative humidity of 60.+ -. 5 ℃ for 12:12 hours for 7 days prior to the experiment. The animal study was approved by the ethics committee of the university of wenzhou medical science and was conducted in accordance with the ARRIVE guidelines.
Mice are free of interfering factors such as infection and inflammation, and are the subjects of the Middle Cerebral Artery Occlusion (MCAO) method. Prior to anesthesia, mice were randomly selected and equally divided into different groups (sham, NS, solvent or MCAO plus treatment group). First, the mice were anesthetized with 1% sodium pentobarbital (0.8 mL/100g; intraperitoneal injection), and then placed in the supine position. Next, after sterilization with iodine, the skin was incised. The Common Carotid Artery (CCA), external Carotid Artery (ECA) and Internal Carotid Artery (ICA) were isolated. Third, ligating CCA and ECA while temporarily closing ICA with arterial clamps. A small incision was then made 4mm from the CCA branch. The MCAO model was created by inserting a wire into the anterior cerebral artery through the CCA. The sham surgery group was treated as such, without ischemia reperfusion.
Neurological scores were assessed 48 hours after MCAO in mice. The results of the five types of kinematic neurological examinations are as follows. 0: no obvious red characters exist; 1: the outer side of the forelimb is difficult to fully extend; 2: inability to extend the outside of the forelimb; 3: winding towards opposite sides; 4: it is difficult or impossible to spontaneously move or scroll. TTC staining was performed to determine infarct size. Firstly, taking out the mouse brain in time, and placing the mouse brain in a refrigerator at the temperature of-20 ℃ for 20min. Next, the brain was cut into 5 pieces, 2mm thick, and placed in TTC staining (Sigma-Aldrich, USA) at 37℃for 30 minutes. TTC-stained brain sections were photographed with a camera and then infarcted areas were treated with Image-Pro plus.
Three separate experiments were performed for each group. All results are expressed as mean ± SD. Statistical differences between groups were determined by student t-test or one-way anova with multiple comparisons in GraphPad Pro (GraphPad, san diego, california, usa).
As shown in FIG. 5, mice in the Vehicle group had a larger cerebral infarction area after CIRI, while mice in the compound 5 treatment group had a decreased cerebral infarction area and a decreased neurological score (Sham). These findings indicate the protective effect of compound 5 on CIRI.
The foregoing disclosure is illustrative of the present invention and is not to be construed as limiting the scope of the invention, which is defined by the appended claims.

Claims (9)

1. A thiobarbituric acid derivative characterized by: which has a chemical structure shown in a general formula I,
The thiobarbituric acid derivative is one of the following compounds:
2. the thiobarbituric acid derivative according to claim 1, wherein: the thiobarbituric acid derivative is one of compounds 1-3 and compounds 5-7.
3. The thiobarbituric acid derivative according to claim 2, characterized in that: the thiobarbituric acid derivative is a compound 5.
4. A process for the preparation of a thiobarbituric acid derivative hybrid according to any one of claims 1 to 3, characterized in that:
the reaction formula is as follows:
5. Use of a thiobarbituric acid derivative according to any one of claims 1-3 for the preparation of a medicament for the treatment of oxidative damage.
6. Use of a thiobarbituric acid derivative according to any one of claims 1-3 for the preparation of Nrf2 activators.
7. Use of a thiobarbituric acid derivative according to any one of claims 1 to 3 for the preparation of a medicament having a function of preventing or treating cerebral ischemia reperfusion injury diseases.
8. A pharmaceutical composition with the function of preventing or treating cerebral ischemia reperfusion injury diseases is characterized by comprising a therapeutically effective amount of active ingredients and pharmaceutically acceptable pharmaceutical excipients; the active ingredient comprising the thiobarbituric acid derivative according to any one of claims 1 to 3 or a pharmaceutically acceptable salt derivative thereof.
9. The pharmaceutical composition having a function of preventing or treating cerebral ischemia reperfusion injury disease according to claim 8, wherein: the pharmaceutical composition has the following preparation forms: injection, tablet, capsule, aerosol, suppository, pellicle, dripping pill, unguent, controlled-release or sustained-release preparation, and nanometer preparation.
CN202410022985.1A 2024-01-08 2024-01-08 Thiobarbituric acid derivative with antioxidation effect, preparation method and application Pending CN118063397A (en)

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