CN104983737B - Pharmaceutical composition and its application in preparing prevention atherosclerosis, dyslipidemia drug - Google Patents

Pharmaceutical composition and its application in preparing prevention atherosclerosis, dyslipidemia drug Download PDF

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CN104983737B
CN104983737B CN201510324417.8A CN201510324417A CN104983737B CN 104983737 B CN104983737 B CN 104983737B CN 201510324417 A CN201510324417 A CN 201510324417A CN 104983737 B CN104983737 B CN 104983737B
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pharmaceutical composition
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atherosclerosis
drug
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CN104983737A (en
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郭守东
田华
秦树存
张颖
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Taishan Medical University
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Abstract

Purposes the invention discloses a kind of pharmaceutical composition and its in prevention atherosclerosis, dyslipidemia and associated disease drug, wherein being administered in combination flavone compound and sialic acid N n acetylneuraminic acid ns in Atherosclerosis Model apo E knock-out mice.Found by experimental study, the pharmaceutical composition can notable regulating lipid metabolism, reduce blood plasma in triglycerides, cholesterol, malonaldehyde and OxLDL ELISA content, promote the artery model mouse of induction high in fat(Apo E knock-out mice)The activity of superoxide dismutase and lipoprotein esterlysis enzyme, reduction lipid is in the accumulation of lipid of apo E knock-out mice liver and the arch of aorta, the effect of improving aortic disease situation, obtain unexpected concertedness antiatherosclerosis.

Description

Pharmaceutical composition and its preparing prevention atherosclerosis, in dyslipidemia drug Application
Technical field
The invention belongs to pharmaceutical technology fields, and it is athero- to prevent artery in particular to pharmaceutical composition and its in preparation Application in hardening, dyslipidemia drug.
Background technology
Cranial vascular disease be a kind of high incidence, high disability rate, high case fatality rate disease, main pathologic process is in blood On the basis of tube wall lesion, blood constituent and(Or)Hemodynamic responses cause ischemic or hemorrhagic disease.Its cause of disease compared with More, one of them important cause of disease is atherosclerosis, and atherosclerosis can cause artery official jargon to narrow, brain tissue occurs Blood supply insufficiency, or even can fall off because of patch, it blocks the cerebrovascular and leads to grave consequences.
The major lesions of atherosclerosis are characterized in lipidosis under the inner membrance at the certain positions of artery, and with smooth muscle The hyperplasia of cell and fibre composition, gradually development form the patch of part, arterial wall also thus thicken and be hardened, inside plaque The lipid binding of necrosis disintegration and deposition, forms the pathological change of athero- substance.Cholesterol and cholesteryl ester are to constitute athero- spot The main component of block.When internal cholesterol is excessively high, especially oxidized low-density lipoprotein(OX-LDL)And triglycerides Increase, then can be swallowed by macrophage, and then be deposited under arterial endothelium, causes subendothelial denaturation, and then blood platelet is caused to exist Wall of Artery is assembled, if simultaneously with arterial wall damage or cholesterol transport obstacle, easily forms fat spot in endarterium, most Afterwards, vascular wall swells, forms atheromatous plaque to official jargon inner process.
Lipoprotein lipase(LPL)It is the key enzyme of lipid-metabolism, is synthesized by parenchyma outside liver and be secreted into blood, hydrolysis breast Rotten particle(chylomicron, CM)And the triglycerides on VLDL(Triglyceride, TG);LPL shortages can cause CM Increase with VLDL levels.
Currently, for the essential therapeutic arsenals of atherosclerosis be use angiogenesis inhibitors and fat-reducing medicament, but It is curative effect is unsatisfactory, and is often accompanied by side effect.Although interventional treatment can make revascularization, restenosis be current Insurmountable difficult medical problem, especially late result have no clear superiority.
Invention content
The present invention is directed to the above shortcoming, a kind of medical composition and its use is provided, experiments have shown that flavonoids Object is closed to combine with sialic acid N-acetyl-neuraminate with significant antiatherosclerosis and Bloodlipid-lowering.
The technical solution adopted by the present invention to solve the technical problems is:A kind of pharmaceutical composition, it is main in pharmaceutical composition It is flavone compound and N-acetyl-neuraminate to want active constituent.
The flavone compound is Quercetin or rutin.
The weight ratio of the flavone compound and N-acetyl-neuraminate is 1:2~4.
Application of the above-mentioned composition in treatment atherosclerosis, dyslipidemia and associated disease drug.
The application of pharmaceutical composition, the daily dosage of each active constituent is Quercetin or rutin in the composition 10-20 mg, N-acetyl-neuraminate 30-40 mg.
The application of pharmaceutical composition, composition are oral preparation.
The dosage form of the application of pharmaceutical composition, the composition is tablet, capsule.
Flavone compound is widely distributed in nature, and is present in the diet of the mankind.Sialic acid is lactation One kind nine carbon sugar in sugar chain non-reducing end in animal body.Research finds that sialic acid can effectively prevent the particles such as albumen in vivo Wall deposits in the blood vessels for agglutination and low-density lipoprotein, and has anti-oxidation function;Its shortage is related to accumulation of lipid.
Compared with prior art, composition of medicine of the present invention significantly reduces the total cholesterol of model mice serum (TC), triglycerides(TG), low-density lipoprotein(LDL-C), malonaldehyde(MDA)And oxidized low-density lipoprotein(OX- LDL), promote serum superoxide dismutases(SOD)With lipoprotein esterlysis enzyme(LPL)Activity;It is to be particularly noted that flavones Class compound combine sialic acid can notable regulating lipid metabolism, reduce serum in TG, TC, LDL-C, MDA content, increase SOD With LPL activity, the lipid accumulation of lipid subendothelial in the artery model mouse liver and the arch of aorta of induction high in fat is reduced, is improved Aortic disease situation, the effect of obtaining good concertedness antiatherosclerosis.
Description of the drawings
Fig. 1 show the influence that drug accumulates liver lipids, compared with blank group, #P < 0.05;##P < 0.01;With Model group is compared, * P < 0.05;* P < 0.01;
Fig. 2 show influence of the drug to arch of aorta accumulation of lipid, compared with blank group, #P < 0.05;##P < 0.01;Compared with model group, * P < 0.05;* P < 0.01.
Specific implementation mode
The present invention will be described in detail in the following with reference to the drawings and specific embodiments:
Embodiment 1
The treatment of Quercetin and N-acetyl-neuraminate to Atherosclerosis Model mouse
SPF grades of apo Es knock out(apoE-/-)Mouse, male, 20 ± 2 g of weight.It gives under experimental conditions common Experimental animal is randomly divided into 5 groups by feed adaptable fed 1 week:Blank group 12, model group 15, Quercetin group 15, N- N acetylneuraminic acid n group 15 and combination A groups 15.In addition to blank group, high lipid food is given(Formula:Cholesterol 1.25%, courage Juice salt 0.5%, lard 10%, yolk powder 10%, basal feed 78.25%);Blank group gives normal diet.After feeding 1 month, respectively Group gives relative medicine in the case where feed is constant, according to 0.2 mL/kg gavages, and blank group and model group give same volume Physiological saline, 1 time a day, successive administration 2 months.The test medicine and dosage of each group are as follows:
Blank group:0.2 mL/kg physiological saline;
Model group:0.2 mL/kg physiological saline;
Quercetin group:10 mg/kg Quercetins;
N-acetyl-neuraminate group:The N-acetyl-neuraminate of 40 mg/kg;
Combine A groups:The N-acetyl-neuraminate of+40 mg/kg of 10 mg/kg Quercetins;
During experiment, each group is without animal dead.In fasting 12h after the last administration, blood vessel eye socket is taken using anticoagulant heparin Venous blood collection 0.8-1.0 mL, yellow Jackets anesthesia are put to death, separated plasma, are saved backup at -80 DEG C;Injection of heart rinses After remaining blood, heart and liver are won rapidly, and whole aorta of the separation arch of aorta to arteria iliaca communis crotch removes blood Fat outside pipe and connective tissue carry out microscopy after dyeing.Gained serum sample detects total cholesterol(TC), triglycerides (TG), low-density lipoprotein white level(LDL-C), hdl level(HDL-C), superoxide dismutase(SOD), oxidation Low-density lipoprotein white level(OX-LDL), malonaldehyde(MDA)With lipoprotein esterlysis enzyme(LPL)Activity.
The experimental result of influence of the drug to plasma TC, TG, LDL-C and HDL-C is as shown in table 1, and model group is relative to sky White control group TC, TG and LDL-C significantly rise, and HDL-C is remarkably decreased;Administration group can lower TC, TG and LDL-C, and on Adjust HDL-C.It is with obvious effects to combine A groups, there is statistical significance relative to model group;N- acetyl ceramide can also significantly reduce TC and TG, and increase HDL-C.
The experimental result table 2 of influence of the drug to blood plasma LPL activity, MDA, OX-LDL and SOD shows, model group relative to Blank control group MDA and OX-LDL level significantly rises, and has statistical significance(P < 0.01), and LPL activity and SOD activity It is remarkably decreased(P < 0.01).It is horizontal that administration group, especially pharmaceutical composition A groups can then significantly reduce MDA and OX-LDL(P < 0.01), raise simultaneously LPL activity and SOD activity(P < 0.01).
Embodiment 2
The treatment of rutin and N-acetyl-neuraminate to Atherosclerosis Model mouse
SPF grades of apo Es knock out(apoE-/-)Mouse, male, 20 ± 2 g of weight.It gives under experimental conditions common Experimental animal is randomly divided into 5 groups by feed adaptable fed 1 week:Blank group 12, model group 12, rutin group 12, N- second Acyl neuraminic acid group 12 and combination B groups 12.In addition to blank group, high lipid food is given(Formula:Cholesterol 1.25%, bile Salt 0.5%, lard 10%, yolk powder 10%, basal feed 78.25%);Blank group gives normal diet.After feeding 1 month, each group In the case where feed is constant, relative medicine is given according to 0.2 mL/kg gavages, blank group and model group give the life of same volume Manage brine, 1 time a day, successive administration 2 months.The test medicine and dosage of each group are as follows:
Blank group:0.2 mL/kg physiological saline;
Model group:0.2 mL/kg physiological saline;
Rutin group:10 mg/kg rutins;
N-acetyl-neuraminate group:The N-acetyl-neuraminate of 30 mg/kg;
Combine B groups:The N-acetyl-neuraminate of+30 mg/kg of 10 mg/kg rutins.
During experiment, each group is without animal dead.In fasting 12h after the last administration, blood vessel eye socket is taken using anticoagulant heparin Venous blood collection 0.8-1.0 mL, yellow Jackets anesthesia are put to death, separated plasma, are saved backup at -80 DEG C;Injection of heart rinses After remaining blood, heart and liver are won rapidly, and whole aorta of the separation arch of aorta to arteria iliaca communis crotch removes blood Fat outside pipe and connective tissue carry out microscopy after dyeing.Gained serum sample detects total cholesterol(TC), triglycerides (TG), low-density lipoprotein white level(LDL-C), hdl level(HDL-C), superoxide dismutase(SOD), oxidation Low-density lipoprotein white level(OX-LDL), malonaldehyde(MDA)With lipoprotein esterlysis enzyme(LPL)Activity.
The experimental result of influence of the drug to plasma TC, TG, LDL-C and HDL-C is as shown in Table 3, and model group is relative to sky White control group TC, TG and LDL-C significantly rise, and HDL-C is remarkably decreased;Administration group can lower TC, TG and LDL-C, and on Adjust HDL-C.It is with obvious effects to combine B groups, there is statistical significance relative to model group;N- acetyl ceramide can also significantly reduce TC and TG, and increase HDL-C.
The experimental result of influence of the drug to blood plasma LPL activity, MDA, OX-LDL and SOD is as shown in Table 4, and model group is opposite It is significantly risen in blank control group MDA and OX-LDL level, there is statistical significance(P < 0.01), and LPL activity and SOD live Property is remarkably decreased(P < 0.01).It is horizontal that administration group, especially pharmaceutical composition B groups can then significantly reduce MDA and OX-LDL(P < 0.01), raise simultaneously LPL activity and SOD activity(P < 0.01).
Embodiment 3
Quercetin and rutin mixture control Atherosclerosis Model mouse with N-acetyl-neuraminate drug combination It treats
SPF grades of apo Es knock out(apoE-/-)Mouse, male, 20 ± 2 g of weight.It gives under experimental conditions common Experimental animal is randomly divided into 5 groups by feed adaptable fed 1 week:Blank group 12, model group 12, mixing Quercetin rutin Group 12, N-acetyl-neuraminate group 12 and combination C groups 12.In addition to blank group, high lipid food is given(Formula:Courage is solid Alcohol 1.25%, bile salt 0.5%, lard 10%, yolk powder 10%, basal feed 78.25%);Blank group gives normal diet.Feed 1 After a month, each group gives relative medicine in the case where feed is constant, according to 0.2 mL/kg gavages, and blank group and model group are given The physiological saline of same volume is given, 1 time a day, successive administration 2 months.The test medicine and dosage of each group are as follows:
Blank group:0.2 mL/kg physiological saline;
Model group:0.2 mL/kg physiological saline;
Mix Quercetin rutin group:+ 10 mg/kg Quercetins of 10 mg/kg Quercetins;
N-acetyl-neuraminate group:The N-acetyl-neuraminate of 40 mg/kg;
Combine C groups:The N-acetyl-neuraminate of+40 mg/kg of+10 mg/kg rutins of 10 mg/kg Quercetins.
During experiment, each group is without animal dead.In fasting 12h after the last administration, blood vessel eye socket is taken using anticoagulant heparin Venous blood collection 0.8-1.0 mL, yellow Jackets anesthesia are put to death, separated plasma, are saved backup at -80 DEG C;Injection of heart rinses After remaining blood, heart and liver are won rapidly, and whole aorta of the separation arch of aorta to arteria iliaca communis crotch removes blood Fat outside pipe and connective tissue carry out microscopy after dyeing.Gained serum sample detects total cholesterol(TC), triglycerides (TG), low-density lipoprotein white level(LDL-C), hdl level(HDL-C), superoxide dismutase(SOD), oxidation Low-density lipoprotein white level(OX-LDL), malonaldehyde(MDA)With lipoprotein esterlysis enzyme(LPL)Activity.
The experimental result of influence of the drug to plasma TC, TG, LDL-C and HDL-C is as shown in Table 5, and model group is relative to sky White control group TC, TG and LDL-C significantly rise, and HDL-C is remarkably decreased;Administration group can lower TC, TG and LDL-C, and on Adjust HDL-C.It is with obvious effects to combine C groups, there is statistical significance relative to model group;N- acetyl ceramide can also significantly reduce TC and TG, and increase HDL-C.
The experimental result of influence of the drug to blood plasma LPL activity, MDA, OX-LDL and SOD is as shown in Table 6, and model group is opposite It is significantly risen in blank control group MDA and OX-LDL level, there is statistical significance(P < 0.01), and LPL activity and SOD live Property is remarkably decreased(P < 0.01).It is horizontal that administration group, especially pharmaceutical composition C groups can then significantly reduce MDA and OX-LDL(P < 0.01), raise simultaneously LPL activity and SOD activity(P < 0.01).
Influence of 1 drug of table to plasma TC, TG, LDL-C and HDL-C
Group TC (mg/dl) TG (mg/dl) LDL-C (mg/dl) HDL-C (mg/dl)
Blank group(n=12) 457.88±45.86 144.91±28.30 115.34±11.29 100.91±19.67
Model group(n=15) 845.93±89.02 ## 195.36±33.41 # 294.14±20.04 ## 74.34±9.58 #
Quercetin group(n=15) 709.87±79.73 160.28±25.59 261.95±47.09 87.60±24.44
N-acetyl-neuraminate group(n=15) 706.23±96.44 * 64.06±21.82 ** 254.55±52.29 * 93.82±19.08 *
Combine A groups(n=15) 563.04±86.17 ** 56.68±15.04 ** 198.15±28.8 0** 116.47±8.30 **
Compared with blank group, #P < 0.05;##P < 0.01;
Compared with model group, * P < 0.05;* P < 0.01;
Influence of 2 drug of table to blood plasma LPL activity, MDA, OX-LDL and SOD
Compared with blank group, #P < 0.05;##P < 0.01;
Compared with model group, * P < 0.05;* P < 0.01;
Influence of 3 drug of table to plasma TC, TG, LDL-C and HDL-C
Group TC (mg/dl) TG (mg/dl) LDL-C (mg/dl) HDL-C (mg/dl)
Blank group(n=12) 491.05±50.17 154.76±32.45 121.42±18.68 97.55±16.24
Model group(n=12) 863.51±76.59 ## 200.05±47.82 # 289.39±26.74 ## 69.16±8.37 #
Rutin group(n=12) 720.36±81.07 * 167.33±19.85 256.36±34.82 * 84.55±17.76 *
N-acetyl-neuraminate group(n=12) 740.34±36.85 * 84.26±30.12 ** 243.77±42.61 * 90.08±11.92 *
Combine B groups(n=12) 607.54±90.26 ** 73.84±22.73 ** 211.08±27.84 * 103.28±11.85 *
Compared with blank group, #P < 0.05;##P < 0.01;
Compared with model group, * P < 0.05;* P < 0.01;
Influence of 4 drug of table to blood plasma LPL activity, MDA, OX-LDL and SOD
Compared with blank group, #P < 0.05;##P < 0.01;
Compared with model group, * P < 0.05;* P < 0.01;
Influence of 5 drug of table to plasma TC, TG, LDL-C and HDL-C
Group TC (mg/dl) TG (mg/dl) LDL-C (mg/dl) HDL-C (mg/dl)
Blank group(n=12) 490.56±65.34 167.35±29.95 128.09±22.37 107.83±21.64
Model group(n=12) 904.50±91.25 ## 216.82±40.70 # 284.98±35.15 ## 76.41±12.77 #
Quercetin and rutin group(n=12) 785.97±81.22 175.04±27.97 261.26±29.33 89.54±20.81*
N-acetyl-neuraminate group(n=12) 756.38±104.47 * 89.66±29.74 ** 244.85±41.78* 90.39±15.35 *
Combine C groups(n=12) 623.49±69.61 ** 76.83±30.48 ** 216.32±38.50 ** 103.76±16.43 **
Compared with blank group, #P < 0.05;##P < 0.01;
Compared with model group, * P < 0.05;* P < 0.01;
Influence of 6 drug of table to blood plasma LPL activity, MDA, OX-LDL and SOD
Compared with blank group, #P < 0.05;##P < 0.01;
Compared with model group, * P < 0.05;* P < 0.01;
Liver organization uses bush uniformly dyeing core, oil red O to contaminate lipid within endothelial cells.Coloration result is in Olympus microscopes (BX51) it is observed under, object lens 10 × combine IPP image collection processing system gathered datas.Experimental result such as Fig. 1 shows, model group Relative to blank group lipid in the subendothelial a large amount of accumulations of the arch of aorta, there is statistical significance(P < 0.01).
The arch of aorta is dyed using oil red O brilliant green, and coloration result is observed under Olympus microscopes (BX51), object lens 10 × combine IPP image collection processing system gathered datas.Experimental result such as Fig. 2 shows, administration group, especially pharmaceutical composition A, B The subendothelial accumulation of lipid of the arch of aorta can be then significantly reduced with C groups;A, B and C group are combined, there is statistical significance(P < 0.01).
Certainly, above description is not limitation of the present invention, and the present invention is also not limited to the example above, this technology neck The variations, modifications, additions or substitutions that the technical staff in domain is made in the essential scope of the present invention should also belong to the present invention's Protection domain.

Claims (5)

1. a kind of prevention atherosclerosis, the pharmaceutical composition of dyslipidemia, which is characterized in that main in described pharmaceutical composition It is flavone compound and N-acetyl-neuraminate to want active constituent, and the flavone compound is Quercetin or rutin.
2. prevention atherosclerosis according to claim 1, the pharmaceutical composition of dyslipidemia, which is characterized in that institute The weight ratio for stating flavone compound and N-acetyl-neuraminate is 1:2-4.
3. application of any compositions of claim 1-2 in preparing prevention atherosclerosis, dyslipidemia drug.
4. the application of pharmaceutical composition according to claim 3, it is characterised in that:Composition is oral preparation.
5. the application of pharmaceutical composition according to claim 4, it is characterised in that:The dosage form of the composition be tablet, Capsule.
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CN110478357A (en) * 2019-07-23 2019-11-22 哈尔滨医科大学 The application of Nitazoxanide and its cylinder metabolism-ure in anti-obesity, reducing blood lipid, anti-fatty liver and antiatherosclerosis
CN113616663A (en) * 2020-05-08 2021-11-09 嘉必优生物技术(武汉)股份有限公司 Use of sialic acid for modulating dyslipidemias caused by abnormal expression of genes responsible for alzheimer's disease
CN113171388A (en) * 2021-04-30 2021-07-27 广州中医药大学(广州中医药研究院) Application of total saponins of astragalus or volatile oil of angelica in preparation of reagent for treating diseases related to abnormal protein expression
CN114272379A (en) * 2021-11-26 2022-04-05 中国科学院生物物理研究所 Novel small molecule inhibitor targeting Ero1 alpha/PDI electron transfer system

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