CN101704866A - Rutin aliphatic ester derivatives, preparation method and application thereof - Google Patents
Rutin aliphatic ester derivatives, preparation method and application thereof Download PDFInfo
- Publication number
- CN101704866A CN101704866A CN200910198424A CN200910198424A CN101704866A CN 101704866 A CN101704866 A CN 101704866A CN 200910198424 A CN200910198424 A CN 200910198424A CN 200910198424 A CN200910198424 A CN 200910198424A CN 101704866 A CN101704866 A CN 101704866A
- Authority
- CN
- China
- Prior art keywords
- rutin
- aliphatic ester
- ester derivatives
- preparation
- hepatitis
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Saccharide Compounds (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention provides rutin aliphatic ester derivatives, which are prepared by chemically synthesizing rutin and the aliphatic ester. The rutin aliphatic ester derivatives have an easily-obtained raw material, low cost, simple preparation method and high yield, and are suitable for industrialized production. The pharmacological test shows that the rutin aliphatic ester derivatives have obvious effect for inhibiting platelet aggregation and inhibiting the activity of hepatitis virus, and can be used for preparing an anti-angiocardiopathy medicine, an anti-cerebrovascular disease medicine, an anti-viral hepatitis B medicine and an anti-hepatitis C medicine.
Description
Technical field
The present invention relates to rutin derivatives, especially relate to rutin aliphatic ester derivatives and preparation method thereof and the application in preparation medicament against cardiovascular disease and medicine resisting viral hepatitis.
Background technology
Along with growth in the living standard, cardiovascular and cerebrovascular diseases becomes healthy significant threat, and China dies from nearly 3,000,000 people of cardiovascular and cerebrovascular diseases every year, accounts for 51% of the annual total dead cause of disease of China.And the patient 75% who survives disability in various degree.Thrombosis, atherosclerosis, inflammatory reaction, ischemic and pour into the pathogenic process of many cardiovascular and cerebrovascular diseases such as radical damage more all (plateletactivating factor, PAF) mediation is closely related with platelet activation factor.
Rutin has effect (Chen Wenmei etc., Chinese combination of Chinese tradiational and Western medicine magazine, 2002,22 (4): 283-285) of antagonism PAF.The pharmacological action of rutin can suppress hematoblastic aggegation, prevents thrombotic effect.Simultaneously the blood vessel injury that can cause medmain, bradykinin increases capillary resistance, reduces capillary permeability, can prevent the oedema that vascular permeability raises and causes.The acute ischemic brain injury there is significant provide protection.National Drug Administration has ratified rutin as drug use, existing rutin tablets and other formulations, the clinical capillary hemorrhage disease that is applicable to that fragility increases also is used for the assisting therapy of hypertensive encephalopathy, hematencephalon, retinal hemorrhage, purpura hemorrhagica, acute hemorrhage nephritis, recidivity nasal bleeding, traumatic pulmonary apoplexy, postpartum hemorrhage etc.
Type B viral hepatitis is a kind of common disease of serious harm human health, and its control is a global public health problem, has caused the concern of countries in the world government.China is the district occurred frequently of viral hepatitis, and average attack rate is 120～1,40/,100,000.Be outstanding with hepatitis B especially wherein.Hepatitis B is a kind of worldwide disease that is caused by hepatitis B virus.Developing country's sickness rate height, according to statistics, the asymptomatic hepatitis B virus carriers in the whole world surpasses 2.8 hundred million, and China accounts for 9,300 ten thousand; Wherein 1/3 clinical manifestation that hepatic injury occurs, there is hepatitis B patient 3,000 ten thousand in China at present, and annual have 300,000 people to die from liver cirrhosis relevant with hepatitis B or liver cancer approximately." the popular present situation of Chinese hepatitis and associated problem analysis report thereof " that China's hepatitis control foundation was announced in 2005 shows only have 19% people accepting antiviral this correct therapeutic modality at present.China hepatitis virus carrier's quantity is just with the speed rapid growth about every year average 2,500,000 people.Hepatitis B is serious harm China people's health not only, but also bring the serious social economic burden to country.The treatment time of hepatitis B is long, and medical expense is higher, adds inappropriate Drug abuse again, has more increased the weight of extra economical load.The direct medical treatment that China is used for the treatment of hepatitis B, liver cirrhosis, liver cancer every year expends and is about 30,000,000,000 yuans.Therefore, strengthening the prevention of viral hepatitis, seek effective methods of treatment of hepatitis B, is the current key subjects that need to be resolved hurrily.Chronic hepatitis B treatment key is an antiviral therapy.
The viral hepatitis C is one of transmissible disease that involves in the whole world, and the whole world has 1.7 hundred million people to infect hepatitis C virus, and China has 3700～4,000 ten thousand infection with hepatitis C virus persons; Its sickness rate has and increases trend year by year, and the Center for Disease Control prediction, from nineteen ninety to 2015 year, have among the population at risk of chronic hepatic diseases, hepatitis C patients will increase by 4 times. in prediction on such basis, if the infection of China's hepatitis C and popular can not being well controlled, to 2015, the situation of the chronic hepatitis B that faced hepatitis C situation and today that we faced is the same severe. therefore, the research of strengthening hepatitis C has importance and urgency. and, owing to lack effective preventative vaccine, control to hepatitis C also will be secular. infection with hepatitis C virus is the major reason of chronic hepatopathy, the infected of about 50% can transfer chronic hepatitis to, the hepatitis C of about 10%-20% developed into liver cirrhosis in 20 years, and with being related closely of hepatocellular carcinoma (HCC). the pathomechanism of hepatitis C and drug research are one of research focuses, but also lack special pathology animal model at present, and also do not have the ideal medicine to treat clinically.
Chinese invention patent (application number is 200810038858.1) discloses the application of rutin in the preparation anti-hepatic-B virus medicine, through hepatitis B surface antigen (HBsAg) and the experiment of hepatitis B virus core antigen (HBeAg) detection reagent, the result shows that rutin can significantly suppress hepatitis B surface antigen (HBsAg) and cAg (HBeAg), has the effect of remarkable inhibition hepatitis B virus.
Though above-mentioned rutin can prepare the medicine of resisting cardiovascular disease and hepatitis B virus resisting, because rutin is water-soluble hardly, is insoluble in fat, oral back is relatively poor in intestinal absorption, and bioavailability is low.Therefore, in order to strengthen the curative effect of medicine, the present invention makes lipophilic rutin aliphatic ester derivatives with rutin.
Summary of the invention
The invention provides rutin aliphatic ester derivatives.
The invention provides the preparation method of this class rutin aliphatic ester derivatives.
The invention provides the application of this class rutin aliphatic ester derivatives in the preparation medicament against cardiovascular disease.
The invention provides the application of this class rutin aliphatic ester derivatives in the preparation medicine resisting viral hepatitis.
The chemical structure of general formula of rutin aliphatic ester derivatives of the present invention is as follows:
In the formula: R
1Be C
2-C
24The acyl group of lipid acid, wherein C
12, C
18The acyl group of saturated fatty acid except.
Rutin aliphatic ester derivatives preparation method's of the present invention step is as follows:
Rutin (Shaanxi Sen Fu Bioisystech Co., Ltd product) and lipid acid are with 1: the mol ratio of 1-8 is dissolved in the solvent tertiary butanol, trimethyl carbinol consumption is 5-10 a times of rutin weight, temperature of reaction 30-100 ℃, the catalyzer lipase Novozym435 (the permanent minister in Shanghai Industrial Co., Ltd. product) that adds 3-8mg by every milliliter of reaction solution, stir, react after 4-20 hour, the 3A molecular sieve by every milliliter of reaction solution adding 50-200mg reacted 82-120 hour.Remove by filter molecular sieve and enzyme, solution decompression evaporate to dryness, residuum are with silica gel column chromatography, and eluent is volume ratio 10-15: ethyl acetate-methyl alcohol mixed liquor of 1, collect product component, and solvent evaporated gets the rutin fatty acid ester, and yield can reach 70-75%.
The pharmacological testing of rutin aliphatic ester derivatives of the present invention
1. with turbidimetry for Determination rabbit washing platelet (WRP) aggregation rate, research rutin aliphatic ester derivatives is to the influence (referring to embodiment 6) of rabbit platelet activation factor inductive platelet aggregation, rutin is the contrast reagent, and the result shows that gathering has very strong restraining effect (referring to table 4) to the rutin fatty acid ester to rabbit PAF induced platelet.
2. test with rabbit body inner injecting and administering, research rutin aliphatic ester derivatives is to the influence (referring to embodiment 7) of Na25-adenosine diphosphate (ADP) disodium (ADP) and arachidonic acid inductive platelet aggregation, and the result shows rutin fatty acid ester anticoagulant (referring to table 5) significantly.
3. it is external to hepatitis B HBsAg and HBeAg restraining effect experiment (referring to embodiment 5) that the employing radioimmunology carries out the rutin aliphatic ester derivatives, the result represents with multiple hole mean value, the result shows that the rutin fatty acid ester has remarkable inhibition hepatitis B virus effect (referring to table 1, table 2 and table 3).
4. carry out hepatitis B virus resisting test in the rutin aliphatic ester derivatives body, after adopting 1 age in days Beijing duck intravenous injection strong positive duck hepatitis B virus, adopt dot hybridization method and radioautograph method, observe the dynamic change (referring to embodiment 8) of duck serum hbv dna (DHBV-DNA).The result shows that the rutin aliphatic ester derivatives has obvious restraining effect (referring to table 6 and table 7) to DHBV-DNA during administration; And after drug withdrawal, do not see " knock-on " phenomenon that obvious hepatitis B virus duplicates again.
5. carry out in the rutin aliphatic ester derivatives body experiment (referring to embodiment 9) to the liver provide protection of bacille Calmette-Guerin vaccine (BCG) and lipopolysaccharides (LPS) induced mice immunological liver injury model.The result shows that the rutin aliphatic ester derivatives can suppress the rising of BCG and LPS inductive mice serum ALT, AST level to some extent, has significant provide protection (referring to table 8) to the mouse immune liver damage.
6. carry out acute toxicity test (referring to embodiment 10) in the rutin aliphatic ester derivatives body.The result shows that the rutin aliphatic ester derivatives has low-down toxicity (referring to table 9).
According to The pharmacological results, the rutin aliphatic ester derivatives demonstrates significant effect in platelet aggregation-against, hepatitis virus resisting activity and liver injury protection experiment, and toxicity is extremely low.Therefore, they can be used as medicament against cardiovascular disease and medicine resisting viral hepatitis is used for animal, are preferred for Mammals, particularly people.
Rutin aliphatic ester derivatives of the present invention can be made medicine by the ordinary method of medication preparation, contains rutin aliphatic ester derivatives and conventional pharmaceutically acceptable carrier of significant quantity in the medicine.With rutin aliphatic ester derivatives and one or more solids or liquid medicine vehicle and (or) assistant agent combines, make the suitable administration form or the dosage form that can be used as drug use, as the preparation of formulations such as tablet, pill, capsule, oral liquid, injection, also can make sustained release preparation, controlled release preparation, targeting preparation and various particulate drug-delivery preparation.The rutin aliphatic ester derivatives that contains 0.1-90 weight % in the preparation.
For tablet is made in the administration unit, can be extensive use of various pharmaceutical carrier well known in the art. about pharmaceutical carrier, for example thinner and absorption agent are as starch, dextrin, calcium sulfate, lactose, N.F,USP MANNITOL, Icing Sugar, sodium-chlor, glucose, urea, lime carbonate, white bole, Microcrystalline Cellulose, pure aluminium silicate etc.; Wetting agent and tackiness agent are as water, glycerine, polyoxyethylene glycol, ethanol, propyl alcohol, starch slurry, dextrin, syrup, Icing Sugar, honey, maltose, liquid glucose, mucialga of arabic gummy, gelatine size, Vltra tears, Xylo-Mucine, lac, methylcellulose gum, potassiumphosphate, polyvinylpyrrolidone etc.; Disintegrating agent is as dry starch, alginates, agar powder, laminaran, yellow soda ash and Citric Acid, lime carbonate, polyoxyethylene sorbitol fatty acid ester, tween-80, gas-producing disintegrant, crosslinked carboxymethyl fecula sodium, sodium laurylsulfonate, methylcellulose gum, ethyl cellulose etc.; The disintegration inhibitor is as sucrose, Tristearoylglycerol, theobroma oil, hydrogenation wet goods; Absorption enhancer is as quaternary ammonium salt, sodium lauryl sulphate etc.; Lubricant is as magnesium laurylsulfate, palmitinic acid, fumaric acid, talcum powder, silicon-dioxide, W-Gum, stearate, boric acid, whiteruss, polyoxyethylene glycol etc.; Sweeting agent is as protein sugar, Sodium Cyclamate, cyclohexane sulfamic acid, alcohol sugar, soluble saccharin (calcium), glycyrrhizin etc.; Correctives, as mentha camphor, various essence, Chinese cassia tree and various fruity material etc. tablet further can also be made coating tablet, for example sugar coated tablet, thin membrane coated tablet, ECT, or double-layer tablets and multilayer tablet. for pill is made in the administration unit, can be extensive use of various pharmaceutical carrier well known in the art. about pharmaceutical carrier, for example thinner and absorption agent are as glucose, lactose, starch, theobroma oil, hydrogenated vegetable oil, polyvinylpyrrolidone, Gelucire, kaolin, talcum powder etc.; Tackiness agent is as gum arabic, tragacanth gum, gelatin, ethanol, honey, syrup, rice paste or batter etc.; Disintegrating agent is as agar powder, dry starch, alginates, sodium laurylsulfonate, methylcellulose gum, ethyl cellulose etc.; Refrigerant, as Liquid Paraffin, Simethicone and plant wet goods. for suppository is made in the administration unit, can be extensive use of various pharmaceutical carrier well known in the art. about this class pharmaceutical carrier theobroma oil for example, the ester of higher alcohols etc. for the administration unit being made capsule, the rutin aliphatic ester derivatives is mixed with above-mentioned various pharmaceutical carriers, and the mixture that will obtain thus places gelatin, gum arabic, methylcellulose gum, sodium alginate, in the hard capsule or soft capsule that polyvinylpyrrolidone and other macromolecular material are made. also the rutin aliphatic ester derivatives can be made microcapsule, be suspended in and form suspensoid in the aqueous medium. for injection preparation is made in the administration unit, solution for example, emulsion, lyophilized injectable powder and suspensoid, can use this area all thinners and emulsifying agent commonly used, as water, ethanol, polyoxyethylene glycol, 1, ammediol, Yelkin TTS, Tweens, the isooctadecanol of ethoxylation, the isooctadecanol of polyoxyization, Polyoxyethylene Sorbitol Fatty Acid Esters etc.; Liposome for example can use methods preparations such as this area membrane process commonly used, reverse phase evaporation, solvent injection method, compound emulsion method. and in addition, ooze injection liquid, can in injection preparation, add proper amount of sodium chloride, glucose or glycerine in order to prepare etc.; In addition, can also add conventional solubility promoter, PH conditioning agent etc. in addition, also can in pharmaceutical preparation, add tinting material, sanitas or other material.
The dosage of rutin aliphatic ester derivatives of the present invention depends on all multifactor, for example will prevent or treat the character and the severity of disease, patient's sex, age, body weight and individual reaction, route of administration and administration number of times etc.Usually about 75 kilograms of patients of body weight, being used for cardiovascular and cerebrovascular diseases, to give the per daily dose of rutin aliphatic ester derivatives be 0.1mg/kg body weight-100mg/kg body weight, preferably 1mg/kg body weight-10mg/kg body weight; Being used for viral hepatitis, to give the per daily dose of rutin aliphatic ester derivatives be 0.001mg/kg body weight-50mg/kg body weight, preferred 0.01mg/kg body weight-10mg/kg body weight.Dosage can the single dose form or be divided into several, for example two, three or four dosage form administrations.Concrete case can be by the doctor according to state of an illness adjustment.
Innovation part of the present invention is:
1. obtain rutin aliphatic ester derivatives of the present invention by chemosynthesis, required cost of material is low, and raw material is easy to get, and its preparation method is easy, and yield is higher, is suitable for suitability for industrialized production.
2. bright by active pharmacology test card, rutin aliphatic ester derivatives of the present invention has remarkable platelet aggregation-against and hepatitis virus resisting activity, and does not see " knock-on " phenomenon that obvious hepatitis B virus duplicates again after the drug withdrawal of rutin aliphatic ester derivatives.
3. rutin is made lipotropy precursor medicine is aliphatic ester derivatives in the present invention, can be dissolved in fat, uses the back can be rapidly by esterase hydrolyzed in blood or histoorgan, and can stop in vivo the longer time.Pharmacological tests shows, rutin before the rutin aliphatic ester derivatives comparison esterification has stronger anticoagulant and the active effect of hepatitis virus, under lower concentration, also show significant inhibitory effect after the esterification, improved specific absorption, thereby strengthened the curative effect of medicine to a great extent.
4. the invention provides the application of rutin aliphatic ester derivatives aspect preparation medicament against cardiovascular disease and medicine resisting viral hepatitis.
5. the medicament against cardiovascular disease and the medicine resisting viral hepatitis of rutin aliphatic ester derivatives preparation of the present invention can be made different dosage form, administration by different way, the selectivity of increase clinical application.
Embodiment
The following examples only are used for further specifying the present invention, but this and do not mean that any limitation of the invention.
Synthesizing of embodiment 1 rutin cetylate
0.1mol rutin and 0.5mol palmitinic acid are dissolved among the trimethyl carbinol 500ml, heating, 60 ℃ of lipase Novozym435 that add 2.5 grams, stir, react after 10 hours, add 50 gram 3A molecular sieves, reacted 96 hours, and removed by filter molecular sieve and enzyme, the solution decompression evaporate to dryness, residuum is with silica gel column chromatography, eluent: ethyl acetate-methyl alcohol mixed liquor (weight ratio 10: 1), collect product component, solvent evaporated, get the rutin cetylate, yield 70%.Following spectral data is shown as the rutin cetylate.
Mp：210-～213℃。Electron spray Zhi Spectrum (ESI-MS) m/z:871.5[M+Na].
1H-NMR (C
5D
5N): δ 7.53 (2H, m, H-2 ' and H-6 '), 6.79 (1H, d, J=9Hz, H-5 '), 6.29 (1H, s, H-8), 6.12 (1H, s, H-6), 5.32 (1H, d, J=7.4Hz, H-1 "), 4.65 (1H, t; J=9Hz, H-4 ' "), 4.48 (1H, s, H-1 ' "), 3.72-2.65 (9H; m, H2 "-H6 ", H2 ' "-H5 ' "), 2.21-2.13 (2H, m, H-α); 1.5 (2H, m, H-β), 1.24-1.13 (32H, m, methene proton in the aliphatic chain).
13C NMR (C
5D
5N): δ 177.5 (C4), 173.5 (C=O, ester carbonyl groups), 168 (C7), 157.5 (C9 or C2), 150.2 (C4 '), 133 (C3), 122.2 (C6 '), 116.5 (C5 "); 77.9 (C3 "), 75.5 (C4 "), 74.7 (C4 ' "), 71.6 (C2 ") 70.8 (C2 ' "), 69.2 (C3 ' "), 67 (C5 ' ").Palmitinic acid part 34.6,31.8,29.6,25.4,25,24.6,23.8,23.1,18.5,15.2,12.1.
Synthesizing of embodiment 2 rutin myristinates
0.1mol rutin and 0.15mol tetradecanoic acid are dissolved among the trimethyl carbinol 500ml, heating, 75 ℃ of lipase Novozym 435 that add 2.8 grams, stir, react after 8 hours, add 40 gram 3A molecular sieves, reacted 90 hours, and removed by filter molecular sieve and enzyme, the solution decompression evaporate to dryness, residuum is with silica gel column chromatography, eluent: ethyl acetate-methyl alcohol mixed liquor (weight ratio 12: 1), collect product component, solvent evaporated, get the rutin myristinate, yield 75%.Following spectral data is shown as the rutin myristinate.
Mp：218-～220℃。ESI-MS?m/z：843.5[M+Na]。
1H-NMR (C
5D
5N): δ 7.52 (2H, m, H-2 ' and H-6 '), 6.78 (1H, d, J=9Hz, H-5 '), 6.27 (1H, s, H-8), 6.14 (1H, s, H-6), 5.34 (1H, d, J=7.4Hz, H-1 "), 4.67 (1H, t; J=9Hz, H-4 ' "), 4.49 (1H, s, H-1 ' "), 3.71-2.63 (9H; m, H2 "-H6 ", H2 ' "-H5 ' "), 2.23-2.13 (2H, m, H-α); 1.45 (2H, m, H-β), 1.22-1.14 (28H, m, methene proton in the aliphatic chain).
13C NMR (C
5D
5N): δ 177.5 (C4), 173.7 (C=O, ester carbonyl groups), 167 (C7), 157.7 (C9 or C2), 150.4 (C4 '), 133.2 (C3), 122.2 (C6 '), 116.4 (C5 "); 77.8 (C3 "), 75.4 (C4 "), 74.6 (C4 ' "), 71.7 (C2 ") 70.9 (C2 ' "), 69.4 (C3 ' "), 67.2 (C5 ' ").Tetradecanoic acid part 34.5,31.7,29.5,25.3,25,24.5,23.8,23.1,18.4,15.3,12.3.
Synthesizing of embodiment 3 rutin linolenates
0.2mol rutin and 0.6mol linolenic acid are dissolved among the trimethyl carbinol 800ml, heating, 70 ℃ of lipase Novozym435 that add 4 grams, stir, react after 10 hours, add 100 gram 3A molecular sieves, reacted 100 hours, and removed by filter molecular sieve and enzyme, the solution decompression evaporate to dryness, residuum is with silica gel column chromatography, eluent: ethyl acetate-methyl alcohol mixed liquor (weight ratio 15: 1), collect product component, solvent evaporated, get the rutin linolenate, yield 71%.Following spectral data is shown as the rutin linolenate.
Mp：202-～204℃。ESI-MS?m/z：893.6[M+Na]。
1H-NMR (C
5D
5N): δ 7.55 (2H, m, H-2 ' and H-6 '), 6.83 (1H, d, J=9Hz, H-5 '), 6.27 (1H, s, H-8), 6.11 (1H, s, H-6), 5.34 (1H, d, J=7.4Hz, H-1 "), 4.61 (1H, t; J=9Hz, H-4 ' "), 4.43 (1H, s, H-1 ' "), 3.71-2.63 (9H; m, H2 "-H6 ", H2 ' "-H5 ' "), 2.25-2.11 (2H, m, H-α); 1.45 (2H, m, H-β), 1.21-1.12 (24H, m, methene proton in the aliphatic chain).
13C NMR (C
5D
5N): δ 177.2 (C4), 173.3 (C=O, ester carbonyl groups), 167.6 (C7), 157.3 (C9 or C2), 150.6 (C4 '), 132.8 (C3), 122.4 (C6 '), 116.7 (C5 "); 77.6 (C3 "), 75.7 (C4 "), 74.6 (C4 ' "), 71.5 (C2 ") 70.6 (C2 ' "), 69.3 (C3 ' "), 66.8 (C5 ' ").Linolenic acid part 122.3,138.5,120.5,137.4,121.3,135,4,34.5,31.7,29.5,25.7,25,24.8,23.5,23.4,18.7,15.3,12.2.
Synthesizing of embodiment 4 DHA-rutin esters
0.2mol rutin and 1.6molDHA are dissolved among the trimethyl carbinol 1000ml, heating, 65 ℃ of lipase Novozym435 that add 5 grams, stir, react after 10 hours, add 100 gram 3A molecular sieves, reacted 96 hours, and removed by filter molecular sieve and enzyme, the solution decompression evaporate to dryness, residuum is with silica gel column chromatography, eluent: ethyl acetate-methyl alcohol mixed liquor (weight ratio 12: 1), collect product component, solvent evaporated, get DHA-rutin ester, yield 72%. following spectral datas are shown as DHA-rutin ester.
Mp：208-～211℃。ESI-MS?m/z：943.6[M+Na]。
1H-NMR (C
5D
5N): δ 7.55 (2H, m, H-2 ' and H-6 '), 6.72 (1H, d, J=9Hz, H-5 '), 6.25 (1H, s, H-8), 6.14 (1H, s, H-6), 5.33 (1H, d, J=7.4Hz, H-1 "), 4.62 (1H, t; J=9Hz, H-4 ' "), 4.43 (1H, s, H-1 ' "), 3.70-2.64 (9H; m, H2 "-H6 ", H2 ' "-H5 ' "), 2.20-2.12 (2H, m, H-α); 1.48 (2H, m, H-β), 1.23-1.12 (20H, m, methene proton in the aliphatic chain).
13C NMR (C
5D
5N) δ 177.1 (C4), 173.2 (C=O, ester carbonyl groups), 167.6 (C7), 157.2 (C9 or C2), 150.1 (C4 '), 132.4 (C3), 122.1 (C6 '), 116.2 (C5 "); 77.5 (C3 "), 75.2 (C4 "), 74.3 (C4 ' "), 71.4 (C2 ") 70.5 (C2 ' "), 69.0 (C3 ' "), 66.7 (C5 ' ").DHA part 127～133 (12C), 34.3,31.5,29.3,25.1,24.6,24.3,23.4,23.1,18.2,15.1,11.8.
The test of embodiment 5 rutin aliphatic ester derivatives hepatitis B virus resisting
1. material and method
1.1 cell cultures and medicine
With the HepG22.2.15 cell is model, with the HepG22.2.15 cell inoculation to 25cm
2In the culturing bottle, every bottle adds DMEM substratum mixed-culture medium (containing 10%FBS and 380 μ g/mlG418) 5ml, puts 37 ℃, 5%CO
2In the constant incubator, went down to posterity in 3～4 days.Use 5%NaHCO
3(DMEM substratum and foetal calf serum are GIBCO company product to adjust pH to 7.2; G418 is a Sigma company product).
Rutin (Shaanxi Sen Fu Bioisystech Co., Ltd product) with dmso solution, is made the solution of 1mg/ml, is diluted to 5 μ g/ml with nutrient solution.
Adefovir ester (Suzhou GlaxoSmithKline PLC pharmaceutical Co. Ltd product) with dmso solution, is made the solution of 1mg/ml, is diluted to 5 μ g/ml with nutrient solution.
The rutin cetylate, the rutin myristinate, rutin linolenate (self-control is referring to embodiment 1,2,3) with dmso solution, is made the solution of 1mg/ml, is diluted to 1 μ g/ml respectively with nutrient solution, 2.5 μ g/ml, the solution of 5 μ g/ml.
The culturing cell Digestive system is with 0.25% trypsin Costar company product) be digested to individual cells, behind the piping and druming mixing, (every porocyte number is 1 * 10 to be inoculated in 96 well culture plates respectively
4) and 24 well culture plates (every porocyte number is 1 * 10
5) on.Add nutrient solution 200ul and 1ml respectively, 37 ℃ of 5%CO
2Incubator is cultivated 48h; After treating cell attachment, abandon culture supernatant, use the nutrient solution that contains different pharmaceutical concentration instead, used the fresh medium that contains equal drug level in per 3 days instead; Experiment was carried out 9 days altogether, and culture supernatant ,-20 ℃ of preservations were collected in the end in the 9th day; Detect HBsAg, HBeAg content in the supernatant liquor with batch test kit.
1.2 experiment grouping
Drug combination adopts the chessboard method design.Be divided into that single to use rutin aliphatic ester derivatives, positive control medicine be adefovir ester, rutin, do not add the negative control group of medicine.Rutin aliphatic ester derivatives concentration is got 1,2.5 and 5 μ g/ml, and positive control medicine adefovir ester concentration is got 5 μ g/ml, rutin Sug/ml.Every group of repeated experiments 3 times.
1.2.1 every hole adds sample 50ul to be measured, establishes each 2 hole of positive and negative contrast, every hole adds each one of negative control (or positive control), and establishes blank one hole;
1.2.2 every hole adds 1 of enzyme conjugates (except the blank hole), abundant mixing, and shrouding is put 37 ℃ and was hatched 30 minutes;
1.2.3 discard liquid in the hole, washings is filled with each hole, leaves standstill 5 seconds, dries, and pats dry after repeating five times;
1.2.4 every hole adds each 1 of developer A liquid, B liquid (zymolyte), abundant mixing, and shrouding is put 37 ℃ and was hatched 15 minutes;
1.2.5 every hole adds 1 of stop buffer, mixing;
1.2.6 use the microplate reader reading, get wavelength 450nm, reference wavelength 630nm reads each hole reading.
1.3
Radioimmunology is adopted in the detection of supernatant liquor HBsAg and HBeAg, and the result represents with multiple hole mean value.
Upright readable HBeAg, HBsAg diagnostic kit (Shanghai Kehua Bio-technology Co., Ltd's product) working method are undertaken by the test kit specification sheets.
Method of calculation: inhibiting rate (%)=((negative control group cpm-experimental group cpm)/negative control group cpm) * 100%
2. result
Relatively use variance analysis between group, adopt SPSS11.0 to finish (seeing Table 1, table 2, table 3).
Table 1 rutin cetylate is to HBsAg and HBeAg restraining effect
Table 2 rutin myristinate is to HBsAg and HBeAg restraining effect
Table 3 rutin linolenate is to HBsAg and HBeAg restraining effect
Above-mentioned experimental result shows that the rutin aliphatic ester derivatives has significant hepatitis B virus restraining effect, and the rutin aliphatic ester derivatives is compared with rutin and had more significant hepatitis B virus restraining effect; Therefore, the rutin aliphatic ester derivatives can be used to prepare the medicine of antiviral hepatitis B.
The test of embodiment 6 rutin cetylate platelet aggregation-againsts
1. medicine and reagent
Rutin (Shaanxi Sen Fu Bioisystech Co., Ltd product) is prepared with dimethyl sulfoxide (DMSO).PAF (Sigma company product), all the other reagent are homemade analytical reagent.Rutin cetylate (self-control is referring to embodiment 1).
Rutin and rutin cetylate are made the solution of 2mmol/L respectively with the DMSO dissolving, are diluted to 0.2mmol/L with physiological saline, 0.4mmol/L, 0.8mmol/L, 1.2mmol/L, the solution of 1.6mmol/L.
2. animal
New Zealand's large ear rabbit, male, body weight (2.0 ± 0.5) kg (The 2nd Army Medical College Experimental Animal Center product) is the qualified animal of one-level.
3. instrument Chrono-log type platelet aggregation instrument.
4. experimental technique
4.1 washing platelet suspension preparation
Rabbit ear arteria comitans nervi mediani is got blood, ACD (Trisodium Citrate 85mmol/L, citric acid 71mmol/L, glucose 110mmol/L) with whole blood with volume ratio anti-freezing in 1: 6, the centrifugal 5min of 30 * g, get supernatant liquor and be rich in the centrifugal 5min of thrombocyte blood plasma (PRP) 500 * g, abandon supernatant, sedimentary thrombocyte is with pH7.4 thrombocyte washings (NaCl300mmol/L, EDTA4mmol/L.Tris5mmol/L) washed twice, at last thrombocyte is suspended in pH7.4TT one bovine serum albumin tyrode's solution [TTBSA solution, KCl2.62mmol/L.MgCl
21.0mmol/L, NaCI137mmol/L, CaCl
21.3mmol/L, Tris10mmol/L, glucose 5.6mmol/L, 0.25% bovine serum albumin (BSA)] in be rabbit washing platelet suspension (WRP).Regulating PC with TTBSA is 2.5 * 10
11/ L.
4.2 the platelet aggregation determination experiment is divided into dimethyl sulphoxide control group, various dose rutin group and various dose rutin cetylate group.Get above-mentioned WRP180ul and add dimethyl sulfoxide (DMSO) respectively, rutin, each 10ul of rutin cetylate solution adds 1.91 * 10
-7Mol/L PAF-0.25%BSA-physiological saline 10ul traces the gathering curve under 37 ℃, calculates platelet aggregation rate and medicine to anticoagulant rate (I), to observe the rutin cetylate to PAF induced platelet accumulative restraining effect.
Inhibiting rate=[(100%-chemical feed pipe platelet aggregation rate)/DMSO manages platelet aggregation rate] * 100%
5. the results are shown in Table 4.
Table 4 rutin cetylate is to PAF induced platelet accumulative influence (f ± s)
P＜0.01.
Above-mentioned experimental result shows that rutin cetylate treated in vitro can obviously suppress PAF (9.55 * 10
-5Mol/L) the inductive rabbit platelet is assembled, and is dose-dependently, IC
50Be 0.12mmol/L.Rutin cetylate group is compared with the rutin group PAF inductive platelet aggregation is had more significant inhibitory effect; Therefore, the rutin aliphatic ester derivatives can be used to prepare the medicine of resisting cardiovascular disease.
Platelet aggregation-against test in the embodiment 7 rutin cetylate bodies
1. the rutin cetylate influences 5-adenosine diphosphate (ADP) disodium (ADP) inductive rabbit platelet accumulative
7 of rabbits, body weight 2.1 ± 0.3kg, male.The rutin cetylate is made liposome: the 20mg/ml intravenous administration, after the administration 10,30,60 and 120min carotid artery get blood, the preparation PRP.The final concentration of ADP is 8mmol/L.Each time is organized all anticoagulant (seeing Table 5) significantly as a result.
2. the rutin cetylate is to the influence of arachidonic acid inductive rabbit platelet congregation
7 of rabbits, body weight 2.1 ± 0.3kg, male.The rutin cetylate is made liposome: 20mg/ml, intravenous administration, after the administration 10,30,60 and 120min carotid artery get blood, the preparation PRP.The final concentration of arachidonic acid (Fhtka product) is 95.5mmol/L.
Each time is organized all anticoagulant (seeing Table 5) significantly as a result.
The quiet notes of table 5 rutin cetylate are to rabbit platelet accumulative restraining effect
x±SD?P＜0.05
Above-mentioned experimental result shows that the rutin cetylate has the obvious suppression effect to 5-adenosine diphosphate (ADP) disodium (ADP) or arachidonic acid inductive platelet aggregation; Therefore, the rutin aliphatic ester derivatives can be used to prepare the medicine of resisting cardiovascular disease.
Hepatitis B virus resisting test in the embodiment 8 rutin cetylate bodies
1. experiment material:
1.1 virus
Duck hepatitis B virus DNA (DHBV-DNA) strong positive serum picks up from the Shanghai sheldrake ,-70 ℃ of preservations.
1.2 animal
1 age in days Beijing duck is planted institute animal rearing field available from Beijing medical courses in general institute medicine.
1.3 medicine
Adefovir ester (Suzhou GlaxoSmithKline PLC pharmaceutical Co. Ltd product); Rutin cetylate (self-control is referring to embodiment 1).Below make the liposome of 1mg/ml respectively, with water for injection dilution back administration.
1.4 reagent
α-32P-dCTP (Beijing Fu Rui company product); Nick translation medicine box (Pu Luomaige company product); SephadexG-50, FicollPVP (Sweden Pharmacia company product); SDS (German Merck company product); Milt DNA, bovine serum albumin (Instite of Biophysics, Chinese Academy of Sciences's product); Nitrocellulose filter 0.45um (An Pu company product).
2. experimental technique
2.1 duplicate the duck hepatitis B virus infection model
Get 30 of 1 age in days Beijing ducks, be divided into 5 groups at random: model control group, positive controls, the high, medium and low dosage group of rutin cetylate.Every group 6.Every through leg shin intravenous injection 0.2ml Shanghai sheldrake DHBV-DNA strong positive duck serum.
2.2 pharmacological agent
Get blood back 7 days of infection from duck leg shin vein, separation of serum, for treating preceding sample (T0) ,-70 ℃ of preservations are to be checked.DHBV infects duckling and carries out the pharmacological agent test after 7 days, and the high, medium and low dosage group of rutin cetylate gives rutin cetylate 10,25 and 50 μ g/kg respectively, and positive controls gives adefovir ester 50 μ g/kg, and model control group gives physiological saline.Gastric infusion, every day 2 times, continuous 10 days, in medication the 5th day (T5), after the 10th day (T10) and the drug withdrawal the 3rd day (P3), get blood from duck leg shin vein, separation of serum ,-70 ℃ of preservations are to be checked.
2.3DHBV-DNA mensuration
The duck serum sample is used
32The P-DHBVDNA probe is made dot hybridization, obtains radioautogram on the x-ray film, measures its OD value in the 490nm place with microplate reader, and dna level is calculated in sxemiquantitative.
3. result
DHBV-DNA total positives behind the duckling infection hepatitis B virus, rutin cetylate 25 μ g/kg, 50 μ g/kg have obvious restraining effect (P＜0.05) to DHBV-DNA, and the rutin cetylate is not seen " knock-on " phenomenon that obvious hepatitis B virus duplicates again after drug withdrawal.The positive drug adefovir ester can be significantly during medication presses down duck DHBV-DNA (P＜0.05), but occurs " knock-on " phenomenon (P＜0.05) that hepatitis B virus duplicates again (see Table 6 and table 7) after the drug withdrawal.
Table 6 rutin cetylate is to the influence (n=6) of duck serum DHBV-DNA level
P＜0.05。
Table 7 rutin cetylate is to the influence (n=6) of the horizontal inhibiting rate of duck serum DHBV-DNA
P＜0.05。
Above-mentioned experimental result shows that the rutin cetylate has significant hepatitis B virus restraining effect; Therefore, the rutin aliphatic ester derivatives can be used to prepare the medicine of antiviral hepatitis B.
In the embodiment 9 rutin linolenate bodies to the experiment of the liver provide protection of bacille Calmette-Guerin vaccine (BCG) and lipopolysaccharides (LPS) induced mice immunological liver injury model
1. experiment material
1.1 laboratory animal
Kunming mouse, ♂, 18～22g (The 2nd Army Medical College Experimental Animal Center product).
1.2 medicine and reagent
Ribavirin injection (Shanghai Xinyi Pharmaceutical Co., Ltd's product).
Bacille Calmette-Guerin vaccine (BCG) (Shanghai biological products company product).
Lipopolysaccharides (LPS) (Sigma company product).
ALT, AST test kit (bio-engineering research institute product is built up in Nanjing)
Rutin linolenate (self-control, referring to embodiment 3) is made the liposome of 1mg/ml, with water for injection dilution back administration.
2. experimental technique
The animal random packet, tail ivBCG2Sit/, the normal control group gives equivalent physiological saline, and the treatment group gives the medicine of various dose, every day 1 time.Positive controls gives ribavirin 50ug/kg, every day 1 time.The normal raising 10 days, after time administration once more tail ivLPS10ug/ only attack, 16h posterior orbit vein is got blood, carries out liver function and detects (Serum ALT, AST measure).
3. result
Rutin linolenate 25,50 μ g/kg have obvious restraining effect (P＜0.05) (seeing Table 8) to BCG and LPS inductive mice serum ALT, the rising of AST level.
Table 8 rutin linolenate is to the influence (n=6) of immunological liver injury mice serum ALT, AST
P＜0.05
Above-mentioned experimental result shows, the rutin linolenate can suppress the rising of BCG and LPS inductive mice serum ALT, AST level to some extent, and the mouse immune liver damage is had provide protection; Therefore, the rutin aliphatic ester derivatives can be used to prepare the medicine of antiviral property third liver.
Embodiment 10 rutin linolenates, rutin cetylate, rutin myristinate acute toxicity test
1. material
1.1 medicine
Rutin (Shaanxi Sen Fu Bioisystech Co., Ltd product) is made the suspension of drug level 0.4g/ml with the 0.5%CMC-Na aqueous solution.
The rutin linolenate is made by oneself, makes the suspension of drug level 0.4g/ml with the 0.5%CMC-Na aqueous solution.
The rutin cetylate is made by oneself, makes the suspension of drug level 0.4g/ml with the 0.5%CMC-Na aqueous solution.
The rutin myristinate is made by oneself, makes the suspension of drug level 0.4g/ml with the 0.5%CMC-Na aqueous solution.
1.2 animal
250 of NIH mouse, male and female half and half, age in 5-6 week, body weight 18-22g (The 2nd Army Medical College Experimental Animal Center product).5 in every cage, the male and female sub-cage rearing observed for 1 week, all healthy survival.
2.1 preliminary experiment
60 of NIH kind mouse, water 12h is can't help in fasting, set up negative control group separately, positive controls (rutin group), experimental group 1 (rutin linolenate), experimental group 2 (rutin cetylate), experimental group 3 (rutin myristinate). positive controls, experimental group is with maximum administration concentration, maximal dose (0.3ml/10g) is once irritated stomach, observes continuously 7 days, dead mouse number and generalized case during the record, if death toll 〉=3O% then carries out the mensuration of medium lethal dose (LD50), if be not enough to cause death, then measure maximum tolerated dose (MTD). negative control group is irritated stomach equivalent tap water, and is surplus with the administration group.
The result: none death of mouse in preliminary experiment, can not survey LD
50So, measure MTD.
2.2 the mensuration of maximum tolerated dose MTD
180 of NIH kind mouse, water 12h is can't help in fasting, is divided into 5 groups at random according to sex, body weight, male and female half and half, 20 every group.Set up negative control group separately, positive controls 1 (rutin group 1), positive controls 2 (rutin group 2), experimental group, control group gives with the amount tap water; Experimental group is with maximum administration concentration, maximal dose (0.3ml/10g) is irritated stomach, wherein positive controls 1 (rutin group 1) is irritated stomach every day 2 times, amounts to dosage and is 24g/kg/d, positive controls 2 (rutin group 2) and irritate stomach every day 3 times, and amounting to dosage is 36g/kg/d.The rutin linolenate, the rutin cetylate, rutin myristinate experiment component is low, high dose group, irritates stomach every day 2 times respectively, and amounting to dosage is 24g/kg/d, irritates stomach every day 3 times, and amounting to dosage is 36g/kg/d.Irritated stomach 1 day, observed continuously 7 days, detail record control group, test group animal ingest every day, the situation reaction and the death condition (seeing Table 9) of drinking-water, ight soil, urine and spirit, four limbs activity aspect.When experiment finishes, all animals is put to death, dissect, visual control main organs lesion tissue situation is calculated MTD.
Table 9 rutin linolenate, rutin cetylate, rutin myristinate acute toxicity test mouse response situation
3. result
With maximum administration concentration, maximum dosage difference gastric infusion, experimental observation is found, after positive controls is irritated stomach the 3rd time, mouse appearance activity reduces, and perpendicular hair, closes abnormal conditions such as order, and symptom continues 7 days, experiment shows that the oral rutin MTD of mouse is lower than 36g/kg/d.After experimental group was irritated stomach the 3rd time, tired contracting and the activity minimizing appearred in the part mouse, but do not have abnormal conditions such as perpendicular hair lazyness is moved, tic, overturning, and be movable as usual behind the 1-3h, occurs fainting from fear tic in the whole process.Put to death animal after 7 days and dissect main organs such as the visual inspection heart, liver, spleen, lung, kidney, the change of outward appearance no abnormality seen.Experiment shows, the oral rutin linolenate of mouse, and the rutin cetylate, rutin myristinate MTD is 36g/kg/d, and the rutin linolenate is described, the rutin cetylate, rutin myristinate toxicity is all very little.
Claims (8)
1. rutin aliphatic ester derivatives is characterized in that having the following chemical structure general formula:
In the formula: R
1Be C
2-C
24The acyl group of lipid acid, wherein C
12, C
18The acyl group of saturated fatty acid except.
2. the preparation method of the described rutin aliphatic ester derivatives of claim 1 is characterized in that preparation method's step is as follows:
Rutin and lipid acid are with 1: the mol ratio of 1-8 is dissolved in the solvent tertiary butanol, trimethyl carbinol consumption is 5-10 a times of rutin weight, temperature of reaction 30-100 ℃, the catalyzer lipase Novozym435 that adds 3-8mg by every milliliter of reaction solution, stir, react after 4-20 hour, 3A molecular sieve by every milliliter of reaction solution adding 50-200mg reacted 82-120 hour, removed by filter molecular sieve and enzyme, the solution decompression evaporate to dryness, residuum is with silica gel column chromatography, and eluent is volume ratio 10-15: ethyl acetate-methyl alcohol mixed liquor of 1, collect product component, solvent evaporated gets the rutin fatty acid ester.
3. by the preparation method of the described rutin aliphatic ester derivatives of claim 2, it is characterized in that lipid acid is C described in preparation method's step
2-C
24Saturated fatty acid, C wherein
12, C
18The acyl group of saturated fatty acid except, or C
2-C
24Unsaturated fatty acids.
4. by the preparation method of the described rutin aliphatic ester derivatives of claim 3, it is characterized in that described C
2-C
24Saturated fatty acid preferably palmitinic acid, tetradecanoic acid; Described C
2-C
24Unsaturated fatty acids preferably linolenic acid, DHA, linolic acid.
5. the application of the described rutin aliphatic ester derivatives of claim 1 is characterized in that the application of described rutin aliphatic ester derivatives in the preparation medicament against cardiovascular disease.
6. the application of the described rutin aliphatic ester derivatives of claim 1 is characterized in that the application of described rutin aliphatic ester derivatives in the preparation medicine resisting viral hepatitis.
7. by the application of the described rutin aliphatic ester derivatives of claim 6, it is characterized in that the application of described rutin aliphatic ester derivatives in the preparation antiviral hepatitis B medicine.
8. by the application of the described rutin aliphatic ester derivatives of claim 6, it is characterized in that the application of described rutin aliphatic ester derivatives in the preparation antiviral property third liver medicine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910198424A CN101704866A (en) | 2009-11-06 | 2009-11-06 | Rutin aliphatic ester derivatives, preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910198424A CN101704866A (en) | 2009-11-06 | 2009-11-06 | Rutin aliphatic ester derivatives, preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101704866A true CN101704866A (en) | 2010-05-12 |
Family
ID=42375125
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200910198424A Pending CN101704866A (en) | 2009-11-06 | 2009-11-06 | Rutin aliphatic ester derivatives, preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101704866A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106755149A (en) * | 2016-11-29 | 2017-05-31 | 大连工业大学 | A kind of method that utilization lipase prepares hydroxytyrosol DHA ester |
| WO2018218903A1 (en) * | 2017-05-27 | 2018-12-06 | 华南理工大学 | Method for preparing troxerutin ester using whole cell catalysis |
-
2009
- 2009-11-06 CN CN200910198424A patent/CN101704866A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106755149A (en) * | 2016-11-29 | 2017-05-31 | 大连工业大学 | A kind of method that utilization lipase prepares hydroxytyrosol DHA ester |
| WO2018218903A1 (en) * | 2017-05-27 | 2018-12-06 | 华南理工大学 | Method for preparing troxerutin ester using whole cell catalysis |
| US11286511B2 (en) | 2017-05-27 | 2022-03-29 | South China University Of Technology | Method for preparing troxerutin ester using whole-cell catalysis |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102675403B (en) | Synthesis of anti-hepatitis B medicine LQC-X and application thereof | |
| US8835134B2 (en) | Cycloastragenol monoglucoside, preparation, pharmaceutical composition and application thereof | |
| CN106794986A (en) | Deuterate or non-deuterate molecule and pharmaceutical preparation | |
| CN104510747B (en) | A kind of new medicine use of iridoid glycoside | |
| Guo et al. | Evaluation of antiviral effect and toxicity of total flavonoids extracted from Robinia pseudoacacia cv. idaho | |
| CN105924492A (en) | Mitochondrion-targeted antitumor pentacyclic triterpene derivatives, and preparation method and application thereof | |
| CN107488162A (en) | A kind of bicyclic alcohol derivatives and its preparation and application | |
| CN104906111B (en) | Pharmaceutical composition prepared from hydroxytyrosol for treating liver injury and preparation process of pharmaceutical composition | |
| CN101704866A (en) | Rutin aliphatic ester derivatives, preparation method and application thereof | |
| WO2018133563A1 (en) | Panax plant extract and pharmaceutical composition and use thereof | |
| CN101723997A (en) | Puerarin glycosylation derivative, medicine compound, preparation method and application thereof | |
| CN101134769A (en) | Ursodeoxycholic acid entecavir acidamide and preparation method and use thereof | |
| CN102068452B (en) | Antiviral medicinal composition | |
| CN105998041B (en) | Rosmarinic acid -4-O- β-D-Glucose glycosides is preparing the application in controlling flu pharmaceutical in advance | |
| CN102755355B (en) | Drug composition resisting influenza virus A or enterovirus | |
| CN101829091B (en) | Use of acetamide dehydrogenation silibinin as medicament for treating viral hepatitis B | |
| CN107056874A (en) | A kind of compound of asiatic acid tenofovir dipivoxil and preparation method thereof | |
| CN103906525A (en) | Application of albizzia chinensis extract in preparation of medicine for treatment of gastric ulcer | |
| CN105348222B (en) | Compound PAC-1 or its salt and the pharmaceutical composition comprising them | |
| CN101829090B (en) | Application of diamine formyl dehydrogenated silybin serving as medicament for curing viral hepatitis B | |
| CN106177035B (en) | Preparation method and application of effective rosa chinensis flower extract with blood sugar reducing and anticancer functions | |
| CN101829086B (en) | Application of aromatic carbamoyl dehydro-silibinin as medicament for treating viral hepatitis B | |
| CN101297861A (en) | Antihypelipidemic active substance extracted from Pagodatree flower | |
| CN101919840B (en) | Application of B/E ring substituted silybin for preparing medicament for treating virus hepatitis B | |
| CN104945525A (en) | Tangut barberry bark fruit polysaccharide, as well as preparation method and use thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20100512 |