CN102670684B - Application of cynanchum atratum total saponin - Google Patents
Application of cynanchum atratum total saponin Download PDFInfo
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- CN102670684B CN102670684B CN201210100072.4A CN201210100072A CN102670684B CN 102670684 B CN102670684 B CN 102670684B CN 201210100072 A CN201210100072 A CN 201210100072A CN 102670684 B CN102670684 B CN 102670684B
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Abstract
The invention discloses an application of a cynanchum atratum total saponin to preparation of an anti-virus medicament. Moreover, the invention further discloses a medicinal composition. In the composition, a therapeutically effective quantity of cynanchum atratum total saponins are taken as active ingredients. The invention discloses a brand new medical application of the cynanchum atratum total saponin. According to the application, further development and utilization of the cynanchum atratum total saponin are facilitated.
Description
Technical field
The invention belongs to the field of Chinese medicines, specifically, is the application for the preparation of antiviral drugs about the scheelite total saponin.
Background technology
The root that Radix Cynanchi Atrati is the upright Radix Cynanchi Atrati of asclepiadaceae Cynanchum plant (Cynanchum atratum Bunge.), Cynanchum versicolor Bunge (Cynanchum versicolor Bunge.).Modern pharmacology research shows, Radix Cynanchi Atrati have significantly bring down a fever, antiinflammatory action (CHINA JOURNAL OF CHINESE MATERIA MEDICA, 1995,20 (12): 751.).
Herpes simplex virus HSV (Herpes simplex virus) belongs to herpetoviridae a virus subfamily, be divided into 1 type and 2 types (HSV-1 and HSV-2) according to antigenic difference, can cause the lip herpes, herpetic keratitis, herpetic dermatitis, Herpes genitalis, kaposi's disease etc., sometimes be also meningitis, the cause of disease of encephalitis, HSV-1 often causes the above infection of waist, especially the infection at eye and position, oral cavity, HSV-1 also can cause genital herpes (10%), HSV-2 is the main pathogens (90%) of genital herpes, can infect neonate by Placenta Hominis and birth canal, cause miscarriage and neonatal death, also relevant with the generation of cervical cancer, endanger larger.
Vesicular stomatitis virus VSV (Vesicular stomatitis virus) is the member of Rhabdoviridae, vesiculovirus genus, is the sub-thread minus-stranded rna virus, can cause the diseases such as herpetic stomatitis.Herpetic stomatitis is multiple mammiferous a kind of acute height contagious disease.With animals such as horse, cattle, pigs, than susceptible, sheep and goat also can infect.The people also occasionally has infection, causes influenza-like symptom, and severe patient can cause encephalitis.Primary disease is classified as the category-A disease by International Office of Epizootics (OIE), at China's herpetic stomatitis, is external zoosis, and country passes in and out in the animal quarantine object and classifies VSV as two class epidemic diseases.
Influenza virus; be called for short influenza virus; that a kind of mankind of causing and animal suffer from grippal RNA viruses; on taxonomy; influenza virus belongs to Orthomyxoviridae family; it can cause acute upper respiratory tract infection, and propagates rapidly by air, all over the world, often has periodically and is very popular.The old man that influenza virus is weak in immunity or the patient of child and some immune disorders can cause more serious symptom, as pneumonia or cardiopulmonary exhaustion etc.Human influenza virus can be divided into 3 classes according to the antigenicity of its nucleoprotein: influenza A virus, Influenza B virus and influenza virus C, wherein WSN virus belongs to influenza A virus.
MHV68 is Mus gamma herpes viruses 68, it is an of great value animal model of research gamma herpes viruses, gamma herpes viruses and some malignant tumor are closely related, as lymphoproliferative disorder after Burkitt ' s lymphoma, nasopharyngeal carcinoma, Hodgkin ' s disease, organ transplantation and the relevant B lymphoma of AIDS.
Summary of the invention
The present inventor finds in the process of the medical usage of the Radix Cynanchi Atrati that studies for a long period of time, and the scheelite total saponin has obvious inhibitory action for multiple virus.
Therefore, primary and foremost purpose of the present invention just is to provide the application of a kind of scheelite total saponin for the preparation of antiviral drugs.
Second purpose of the present invention is to provide a kind of pharmaceutical composition that the scheelite total saponin is active component of take.
According to the present invention, described scheelite total saponin can be used for preparing antiviral drugs.According to a preferred embodiment of the invention, described virus comprises: herpes simplex virus, vesicular stomatitis virus, influenza virus, and Mus gamma herpes viruses 68.Wherein, described herpes simplex virus comprises HSV-1 type and HSV-2 type virus.
Antiviral medicinal composition of the present invention take the treatment effective dose the scheelite total saponin be active component.
According to the present invention, described pharmaceutical composition also comprises one or more pharmaceutically suitable carrier.Preferably, described pharmaceutically suitable carrier is selected from: soft phospholipid, aluminium stearate, aluminium oxide, ion exchange material, self-emulsifying drug delivery system, tween or other surfactants, serum albumin, buffer substance as phosphate, glycine, sorbic acid, water, salt, electrolyte as sulfate protamine, sodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salt, magnesium silicate and satisfied fatty acid partial glycerol ester admixture.
According to the present invention, described pharmaceutical composition can also comprise one or more excipient substances.Preferably, described excipient substance is selected from: binding agent, filler, disintegrating agent, lubricant, absorption enhancer, absorption carrier, flavouring agent, sweeting agent, excipient, diluent and wetting agent.
The present invention has opened up the medical usage of a kind of brand-new Radix Cynanchi Atrati, contributes to the further exploitation of Radix Cynanchi Atrati.
The specific embodiment
Below in conjunction with specific embodiment, preparation method and the medical usage of scheelite total saponin of the present invention is described in further detail.Should be understood that following examples are only for the present invention is described but not for limiting scope of the present invention.
The inventor is engaged in the medical usage research of Radix Cynanchi Atrati for a long time, in long-term research, finds, the scheelite total saponin all has obvious inhibitory action for multiple virus, thereby can be for the preparation of antiviral drug.These viruses comprise: herpes simplex virus (HSV-1 type and HSV-2 type), vesicular stomatitis virus (VSV), influenza virus (WSN), and Mus gamma herpes viruses 68 (MHV68).
Scheelite total saponin of the present invention can be used separately or use with the form of pharmaceutical composition.Pharmaceutical composition comprises scheelite total saponin of the present invention and the pharmaceutically suitable carrier as active component.
Wherein, " treatment effective dose " refers to the amount of the extract that can reach therapeutic effect.One of skill in the art can understand " treatment effective dose " and can and may share etc. with the other treatment agent and different along with the use of the mode of administration, carrier.
" pharmaceutically suitable carrier " can not destroy the pharmaceutical active of scheelite total saponin of the present invention, its effective dose simultaneously, and can bringing into play pharmaceutical carrier, to make the consumption of used time nontoxic to human body.
Described pharmaceutically suitable carrier includes but not limited to: soft phospholipid, aluminium stearate, aluminium oxide, ion exchange material, self-emulsifying drug delivery system, tween or other surfactants, serum albumin, buffer substance as phosphate, glycine, sorbic acid, water, salt, electrolyte as sulfate protamine, sodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salt, magnesium silicate, satisfied fatty acid partial glycerol ester admixture etc.
Compositions of the present invention can further include one or more excipient substances, and excipient substance commonly used is as binding agent (as microcrystalline Cellulose), filler (as starch, glucose, Lactis Anhydrous and lactose beadlet), disintegrating agent (as cross-linked pvp, crosslinked carboxymethyl fecula sodium, cross-linking sodium carboxymethyl cellulose, low-substituted hydroxypropyl cellulose), lubricant (as magnesium stearate) and absorption enhancer, absorption carrier, flavouring agent, sweeting agent, excipient, diluent, wetting agent etc.
The place of production of the Radix Cynanchi Atrati of using in following examples is Henan.
Related percentage ratio (%) in following examples, except special instruction, be mass percent.
The preparation of embodiment 1, scheelite total saponin
Get Radix Cynanchi Atrati medicinal material coarse powder 200g, add 10 times of amount alcohol reflux of 70% 3 times, 2h/ time, filtration.Merging filtrate, filtrate decompression is concentrated into every milliliter and is equivalent to the 0.5g crude drug, and concentrated solution centrifugal (3000r/min, 15min) obtains supernatant, again supernatant is added on macroporous adsorbent resin (AB8, D101, the HP20 etc.) post of 6 times of amounts, the loading flow velocity is 6BV/h, first with 6 column volumes of distilled water, more successively with 6 times of column volumes, 30%, 90% ethanol gradient elution, collect eluent, pressurization is concentrated appropriate, and the lyophilization of then reducing pressure, to dry, is pulverized to such an extent that fine powder is scheelite total saponin sample.Wherein the scheelite total saponin content is more than 50%.
The scheelite total saponin, the brown color powder, the Lieberman-Buchard reaction is positive.
Embodiment 2, the scheelite total saponin inhibitory action to HSV-1 virus
Experiment material:
Cell: African green monkey kidney cell (Vero), purchased from Chinese Academy of Sciences's cell bank.
Virus: HSV-1-GFP.
Tested medicine: scheelite total saponin (sample of embodiment 1 preparation), be dissolved in DMSO, be prepared into the 60mg/mL mother solution.
Experimental technique:
The Vero cell is cultivated according to a conventional method, is inoculated in 96 orifice plates next day, after cell attachment, virus infected cell, after 3 hours, remove containing virus-culturing fluid, process cell, fluorescence microscopy Microscopic observation GFP protein expression situation after 22 hours with certain density scheelite total saponin.
Experimental result is as shown in the following Table 1:
The inhibition to HSV-1 virus of table 1, scheelite total saponin
Above experiment shows, in African green monkey kidney cell, the scheelite total saponin has quite significantly inhibitory action to HSV-1, and the scheelite total saponin is dose-dependence to the inhibition of HSV-1.
Embodiment 3, the scheelite total saponin inhibitory action to HSV-2 virus
Experiment material:
Cell: African green monkey kidney cell (Vero), purchased from Chinese Academy of Sciences's cell bank.
Virus: herpes simplex virus HSV-2 type Sm333 standard strain, by Beijing, Inst. of Medicinal Biological Technology, Chinese Academy of Medical Sciences provides.
Tested medicine: scheelite total saponin (sample of embodiment 1 preparation), be dissolved in DMSO, be prepared into the 60mg/mL mother solution.
Experimental technique:
The Vero cell is cultivated according to a conventional method, next day, be inoculated in 96 orifice plates, and monolayer culture is after 24 hours, supernatant, washing are abandoned in suction, add virus liquid, after adsorbing 2 hours, suck virus, the medicinal liquid that adds variable concentrations, cultivate after 72 hours and observe CPE (pathological changes), establish cell matched group, virus control group, positive controls, Experimental agents group, simultaneously observation of cell pathological changes situation.
Experimental result is as shown in the following Table 2:
The inhibition to HSV-2 virus of table 2, scheelite total saponin
Above experiment shows, in the Vero cell, the scheelite total saponin has obvious inhibitory action to HSV-2 virus, and the scheelite total saponin is dose-dependence to the inhibition of HSV-2.
Embodiment 4, the scheelite total saponin inhibitory action to VSV virus
Experiment material:
Cell: human lung adenocarcinoma cell (A549), purchased from Chinese Academy of Sciences's cell bank.
Virus: VSV-GFP.
Tested medicine: scheelite total saponin (sample of embodiment 1 preparation), be dissolved in DMSO, obtain the 60mg/mL mother solution.
Experimental technique:
The A549 cell is cultivated according to a conventional method, is inoculated in 96 orifice plates next day, after cell attachment, virus infected cell, after 1 hour, remove containing virus-culturing fluid, process cell, fluorescence microscopy Microscopic observation GFP protein expression situation after 18 hours with certain density scheelite total saponin.
Experimental result is as shown in the following Table 3:
Table 3, the scheelite total saponin inhibitory action to VSV virus
The experimental result of table 3 shows, in human lung adenocarcinoma cell, the scheelite total saponin has obvious inhibitory action to VSV virus, and the scheelite total saponin is dose-dependence to the inhibition of VSV.
Embodiment 5, the scheelite total saponin inhibitory action to WSN virus
Experiment material:
Cell: HEKC 293T, purchased from ATCC.
Virus: WSN.
Tested medicine: scheelite total saponin (sample of embodiment 1 preparation), be dissolved in DMSO, obtain the 60mg/mL mother solution.
Experimental technique:
The 293T cell is incubated at according to a conventional method containing in the DMEM of 10% hyclone, is inoculated in 96 orifice plates next day, after cell attachment, and virus infected cell, after 1 hour, remove containing virus-culturing fluid, respectively with drug treating 1,2, after 4 hours, be replaced with fresh culture, observed result after 8 hours.Experimental result is as shown in the following Table 4:
Table 4, the scheelite total saponin inhibitory action to WSN
The experimental result of above table 4 shows, the scheelite total saponin has dose-dependent effect to the inhibition of WSN virus, and prolongation in time of its suppression efficiency and increasing.
Embodiment 6, the scheelite total saponin inhibitory action to MHV68 virus
Experiment material:
Cell: HEKC 293T, purchased from ATCC.
Virus: MHV68.
Tested medicine: scheelite total saponin (sample of embodiment 1 preparation), be dissolved in DMSO, obtain the 60mg/mL mother solution.
Experimental technique:
The 293T cell is incubated at according to a conventional method containing in the DMEM of 10% hyclone, be inoculated in 96 orifice plates next day, after cell attachment, process in two ways cell: 1, the drug effect cell is after 1 hour, remove medicinal liquid, add viral infection observed result (1HBI) after 1 hour; 2, virus infected cell, after 1 hour, is removed containing virus-culturing fluid, and drug treating is observed result (1HPI) after 1 hour.
Experimental result is as shown in the following Table 5:
Table 5, the scheelite total saponin inhibitory action to MHV68
The experimental result of above table 5 shows, the scheelite total saponin has good inhibition to MHV68 virus, and the model of action difference, and its inhibition is also different, first infects virus treated with medicaments again, higher to viral suppression ratio.
In sum, the scheelite total saponin all has good inhibitory action for multiple virus, can be used for preparing antiviral drugs, perhaps further make antiviral medicinal composition, except take the treatment effective dose the scheelite total saponin as active component, described pharmaceutical composition also comprises one or more pharmaceutically suitable carrier, and described pharmaceutically suitable carrier can be selected conventional carrier as known in the art; In addition, described pharmaceutical composition can also comprise the excipient substance that one or more are commonly used, and this is apparent for a person skilled in the art.
Claims (8)
1. the application of scheelite total saponin, is characterized in that, for the preparation of antiviral drugs.
2. application as claimed in claim 1, is characterized in that, described virus comprises herpes simplex virus, vesicular stomatitis virus, influenza virus, and Mus gamma herpes viruses 68.
3. application as claimed in claim 2, is characterized in that, described herpes simplex virus comprises HSV-1 type and HSV-2 type virus.
4. an antiviral medicinal composition, is characterized in that, described pharmaceutical composition take the treatment effective dose the scheelite total saponin be active component.
5. pharmaceutical composition as claimed in claim 4, is characterized in that, also comprises one or more pharmaceutically suitable carrier.
6. pharmaceutical composition as claimed in claim 5, it is characterized in that, described pharmaceutically suitable carrier is selected from: soft phospholipid, aluminium stearate, aluminium oxide, ion exchange material, self-emulsifying drug delivery system, tween or other surfactants, serum albumin, buffer substance as phosphate, glycine, sorbic acid, water, salt, electrolyte as sulfate protamine, sodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salt, magnesium silicate and satisfied fatty acid partial glycerol ester admixture.
7. pharmaceutical composition as claimed in claim 4, is characterized in that, also comprises one or more excipient substances.
8. pharmaceutical composition as claimed in claim 7, is characterized in that, described excipient substance is selected from: binding agent, filler, disintegrating agent, lubricant, absorption enhancer, absorption carrier, flavouring agent, sweeting agent, excipient, diluent and wetting agent.
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| CN107184886B (en) * | 2017-05-16 | 2018-06-19 | 贾洪章 | Anti virus herb composition and its preparation method and application |
| CN111358804B (en) * | 2020-04-20 | 2021-02-02 | 贵州省中国科学院天然产物化学重点实验室(贵州医科大学天然产物化学重点实验室) | Application of Cynoside H in preparation of medicine for preventing and treating breast cancer |
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| 大孔吸附树脂法富集白薇中总皂苷的工艺研究;吕永海，等;《中国中药杂志》;20080630;第33卷(第12期);第1390-1393页 * |
| 抗Ⅱ型单纯疱疹病毒的中草药研究;周元，等;《江西医学院学报》;19881231;第28卷(第4期);第1-7页 * |
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Owner name: ZHEJIANG ZHONGKE HONGSHOU PHARMACEUTICAL TECHNOLOG Free format text: FORMER OWNER: BEIJING COSCI MED-TECH CO., LTD. Effective date: 20150707 |
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