CN102526168B - Micro-capsules of anti-influenza-virus effective part of phyllanthus urinaria, and preparation method and application thereof - Google Patents
Micro-capsules of anti-influenza-virus effective part of phyllanthus urinaria, and preparation method and application thereof Download PDFInfo
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- CN102526168B CN102526168B CN201210039716.3A CN201210039716A CN102526168B CN 102526168 B CN102526168 B CN 102526168B CN 201210039716 A CN201210039716 A CN 201210039716A CN 102526168 B CN102526168 B CN 102526168B
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- influenza virus
- cacumen securinegae
- securinegae suffruticosae
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Abstract
The invention discloses micro-capsules of an anti-influenza-virus effective part of phyllanthus urinaria, and a preparation method and application thereof. The preparation method comprises the following steps of: heating and extracting phyllanthus urinaria medicinal materials by using 30 to 80 percent ethanol, recovering the ethanol from an extracting solution, adsorbing by using a macroporous resin, eluting by using 50 to 90 percent ethanol, recovering the ethanol, and extracting the effective part by using dichloromethane, ethyl acetate or n-butyl alcohol; mixing capsule materials, emulsifier, an antisticking agent and a solvent to obtain a mixed solution, adding an aqueous solution of the effective part of the phyllanthus urinaria into the mixed solution, performing ultrasonic emulsification, allowing the mixture to pass through a microfiltration membrane, and spray-drying to obtain the micro-capsules. The micro-capsules can kill influenza virus and can be used for anti-influenza-virus medicines or disinfection of epidemic prevention articles for human or animals. The micro-capsules improve the stability of the effective part of the phyllanthus urinaria and improve the action effect of the effective part obviously.
Description
Technical field
The present invention relates to medical manufacture field, be specifically related to a kind of Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule preparation method and as the application of antiviral drugs.
Background technology
Cacumen Securinegae Suffruticosae (Phyllanthus urinaria L) is Euphorbiaceae phyllanthus plant, and its cool in nature, micro-hardship is sweet, has the effect of suppressing the hyperactive liver, annealing, improving eyesight, removing toxic substances and promoting subsidence of swelling, conjunctival congestion and swelling pain, infantile malnutrition, dysentery, the venom etc. of being used for the treatment of among the people.Since India scholar Thyagarajan has reported Cacumen Securinegae Suffruticosae, can make after 59% anti-hepatitis B surface antigen, its anti-HBV effect has caused domestic and international broad research.Research mainly for Cacumen Securinegae Suffruticosae is its antitumor action and anti-HBV effect at present.
Recently find that Cacumen Securinegae Suffruticosae has certain curative effect in anti-other viruses.The effect that institute of materia medica, Shandong has In Vitro Anti Herpes simplex virus from dehydrogenation chebulinic acid methyl ester and the Methy Brevifolin-carboxlate of Cacumen Securinegae Suffruticosae separation (Li Renjiu. the lignan component phyllanthus plant. external medicine (plant amedica fascicle), 1997, 12 (3): 99～100), Guo Weidong etc. have reported that Cacumen Securinegae Suffruticosae has the effect (Guo Weidong of In Vitro Anti herpes simplex virus II type, Deng Xuelong, Dong Baizhen. the effect of the external herpes simplex virus type II of phyllanthus plant. Traditional Chinese Medicine University Of Guangzhou's journal, 2000, 17 (1): 54～57), CN1082377C has reported that Cacumen Securinegae Suffruticosae has the effect of anti HIV-1 virus.Wei body eel etc. reported Cacumen Securinegae Suffruticosae have the effect of anti-new castle disease virus (Wei body eel, Yu Jie, Yu Sijiu. the screening of Cacumen Securinegae Suffruticosae anti-new castle disease virus active component. Agriculture of Anhui science, 2009,37 (18): 8505～8506).
Relevant Cacumen Securinegae Suffruticosae anti-influenza virus activity component preparation and preparation thereof there is no report.
Summary of the invention
The shortcoming that the object of the invention is to overcome prior art, with not enough, provides a kind of preparation method and application thereof of Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule.The method is the effective site of having extracted Cacumen Securinegae Suffruticosae resisiting influenza virus, and makes microcapsule, then makes various pharmaceutical dosage forms.Keep the stability of Cacumen Securinegae Suffruticosae resisiting influenza virus composition, brought into play better action effect.
Object of the present invention is achieved through the following technical solutions:
A preparation method for Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule, comprises the steps:
(1) to the ethanol that adds 5～10 times of its weight, mass fraction 30～80% in Cacumen Securinegae Suffruticosae, 30～80 ℃ of heating extraction 1～3 hour, filter, filtering residue repeats to extract 1～2 time with above-mentioned solvent again, filter, merging filtrate, in 30～60 ℃ of vacuum concentration to 1/3～2/3 of original volume, adds macroporous resin adsorption 8～24 hours; Take out macroporous resin, with mass fraction 50～90% ethanol or acetone eluting, 30～60 ℃ of vacuum concentration of eluent are to 1/3 of original volume～2/3 o'clock, with equal-volume dichloromethane, ethyl acetate or n-butanol extraction 1～5 time, combining extraction liquid, 30～60 ℃ of vacuum concentration are also dried 3～6 hours, obtain Cacumen Securinegae Suffruticosae resisiting influenza virus effective site;
(2) capsule material is mixed with to the capsule material solution of 1～10% mass fraction with solvent, add the emulsifying agent of capsule material solution 0.5～5% mass fraction, the Cacumen Securinegae Suffruticosae resisiting influenza virus effective site of the antiplastering aid of 0.1～1% mass fraction, 1～20% mass fraction, ultrasonic emulsification 0.5～2 hour, cross microporous filter membrane, spraying is dry, makes Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule.
Preferably, the described macroporous resin of step (1) is D101, DM130, HPD600, AB-8, XAD-16 or NkA-9.
Preferably, the described capsule material of step (2) is one or more in acrylic resin, hydroxypropyl methylcellulose, ethyl cellulose, methylcellulose, gelatin, arabic gum, cyclodextrin, chitosan.
Preferably, the described solvent of step (2) is one or more in methanol, ethanol, acetone, isopropyl alcohol, dichloromethane.
Preferably, the described emulsifying agent of step (2) is non-ionic surface active agent.
Preferably, described emulsifying agent is one or more in monoglyceride, Tween 80, sorbester p18, sucrose fatty acid ester and poloxamer.
Preferably, the described antiplastering aid of step (2) is one or more in Polyethylene Glycol, Talcum, silica gel and magnesium stearate.
The application of described Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule, is made into any of external, oral or injecting drug use dosage form, be made into people with or brutish for resisiting influenza virus or disinfectant anti-epidemic product.
The present invention has following advantage and effect with respect to prior art:
(1) the present invention has found Chinese medicine Cacumen Securinegae Suffruticosae resisiting influenza virus effective site, and by it separation and purification, for Cacumen Securinegae Suffruticosae application provides new purposes.
(2) microcapsule formulation of the present invention, overcomes that Cacumen Securinegae Suffruticosae resisiting influenza virus composition is unstable, fugitive, the shortcoming of weak effect, has increased it and has been utilized degree by body, has brought into play better Cacumen Securinegae Suffruticosae composition action effect.
(3) this preparation method is simple, is applicable to suitability for industrialized production.
The specific embodiment
Below in conjunction with embodiment, the present invention is described in further detail, but embodiments of the present invention are not limited to this.
Embodiment 1
(1) Cacumen Securinegae Suffruticosae 1kg, adds 5kg, 30% ethanol, and 80 ℃ of heating extraction 3 hours are filtered, and filtering residue repeats to extract 2 times with above-mentioned solvent again, filtration, and 60 ℃ of vacuum concentration of merging filtrate, to 2/3 of original volume, add D101 macroporous resin adsorption 24 hours; Take out macroporous resin 90% ethanol elution, 50 ℃ of vacuum concentration of eluent, to 1/3 o'clock of original volume, are used equal-volume dichloromethane extraction 1 time, combining extraction liquid, and 30 ℃ of vacuum concentration are also dried 3 hours, obtain Cacumen Securinegae Suffruticosae resisiting influenza virus effective site 55g.
(2) by acrylic resin IV100g, with dichloromethane, be mixed with the capsule material solution of 5% mass fraction, add capsule material liquid quality fraction 0.5% monoglyceride, 0.1% Polyethylene Glycol, 20% Cacumen Securinegae Suffruticosae resisiting influenza virus effective site, ultrasonic emulsification 0.5 hour, cross microporous filter membrane, spraying is dry, makes Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule.
Embodiment 2
(1) Cacumen Securinegae Suffruticosae 1kg, adds 30 ℃ of heating extraction of 10kg 80% ethanol 1 hour, filters, and filtering residue repeats to extract 1 time with above-mentioned solvent again, filters, and 30 ℃ of vacuum concentration of merging filtrate, to 1/3 of original volume, add DM130 macroporous resin adsorption 8 hours; Take out macroporous resin 50% ethanol elution, 60 ℃ of vacuum concentration of eluent were to 2/3 o'clock of original volume, and with equal-volume ethyl acetate extraction 3 times, combining extraction liquid, 50 ℃ of vacuum concentration are also dried 3 hours, obtain Cacumen Securinegae Suffruticosae resisiting influenza virus effective site 102g.
(2) by ethyl cellulose 100g, with acetone, be mixed with the capsule material solution of 10% mass fraction, add capsule material liquid quality fraction 5% Tween 80,0.5% Talcum, 10% Cacumen Securinegae Suffruticosae resisiting influenza virus effective site, ultrasonic emulsification 2 hours, cross microporous filter membrane, spraying is dry, makes Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule.
Embodiment 3
(1) Cacumen Securinegae Suffruticosae 1kg, adds 60 ℃ of heating extraction of 8kg 50% ethanol 2 hours, filters, and filtering residue repeats to extract 1 time with above-mentioned solvent again, filters, and 50 ℃ of vacuum concentration of merging filtrate, to 2/3 of original volume, add HPD600 macroporous resin adsorption 12 hours; Take out macroporous resin 70% ethanol elution, 50 ℃ of vacuum concentration of eluent, to 1/3 o'clock of original volume, are used equal-volume n-butanol extraction 2 times, combining extraction liquid, and 60 ℃ of vacuum concentration are also dried 5 hours, obtain Cacumen Securinegae Suffruticosae resisiting influenza virus effective site 83g.
(2) by hydroxypropyl cellulose 100g, with ethanol, be mixed with the capsule material solution of 8% mass fraction, add capsule material liquid quality fraction 3% sorbester p18,1% silica gel, 5% Cacumen Securinegae Suffruticosae resisiting influenza virus effective site, ultrasonic emulsification 1 hour, cross microporous filter membrane, spraying is dry, makes Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule.
Embodiment 4
(1) Cacumen Securinegae Suffruticosae 1kg, adds 60 ℃ of heating extraction of 6kg 60% ethanol 2 hours, filters, and filtering residue repeats to extract 2 times with above-mentioned solvent again, filters, and 60 ℃ of vacuum concentration of merging filtrate, to 2/3 of original volume, add AB-8 macroporous resin adsorption 16 hours; Take out 80% acetone eluting for macroporous resin, 40 ℃ of vacuum concentration of eluent, to 1/3 o'clock of original volume, are used equal-volume dichloromethane extraction 2 times, combining extraction liquid, and 40 ℃ of vacuum concentration are also dried 4 hours, obtain Cacumen Securinegae Suffruticosae resisiting influenza virus effective site 70g.
(2) by arabic gum 100g, with isopropyl alcohol, be mixed with the capsule material solution of 2% mass fraction, add capsule material liquid quality fraction 1% sucrose fatty acid ester, 0.2% magnesium stearate, 3% Cacumen Securinegae Suffruticosae resisiting influenza virus effective site, ultrasonic emulsification 2 hours, cross microporous filter membrane, spraying is dry, makes Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule.
Embodiment 5
(1) Cacumen Securinegae Suffruticosae 1kg, adds 40 ℃ of heating extraction of 7kg 70% ethanol 3 hours, filters, and filtering residue repeats to extract 1 time with above-mentioned solvent again, filters, and 40 ℃ of vacuum concentration of merging filtrate, to 2/3 of original volume, add XAD-16 macroporous resin adsorption 10 hours; Take out 60% acetone eluting for macroporous resin, 30 ℃ of vacuum concentration of eluent were to 1/3 o'clock of original volume, and with equal-volume ethyl acetate extraction 5 times, combining extraction liquid, 50 ℃ of vacuum concentration are also dried 3 hours, obtain Cacumen Securinegae Suffruticosae resisiting influenza virus effective site 85g.
(2) by cyclodextrin 100g, with methanol, be mixed with the capsule material solution of 1% mass fraction, add capsule material liquid quality fraction 1.5% poloxamer, 0.5% Talcum, 1% Cacumen Securinegae Suffruticosae resisiting influenza virus effective site, ultrasonic emulsification 1 hour, cross microporous filter membrane, spraying is dry, makes Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule.
Embodiment 6
(1) Cacumen Securinegae Suffruticosae 1kg, adds 70 ℃ of heating extraction of 9kg 40% ethanol 2 hours, filters, and filtering residue repeats to extract 1 time with above-mentioned solvent again, filters, and 60 ℃ of vacuum concentration of merging filtrate, to 2/3 of original volume, add NKA-9 macroporous resin adsorption 20 hours; Take out 50% acetone eluting for macroporous resin, 30 ℃ of vacuum concentration of eluent, to 1/3 o'clock of original volume, are used equal-volume n-butanol extraction 4 times, combining extraction liquid, and 60 ℃ of vacuum concentration are also dried 4 hours, obtain Cacumen Securinegae Suffruticosae resisiting influenza virus effective site 97g.
(2) by chitosan 100g, with ethanol, be mixed with the capsule material solution of 3% mass fraction, add capsule material liquid quality fraction 2% poloxamer, 0.8% silica gel, 12% Cacumen Securinegae Suffruticosae resisiting influenza virus effective site, ultrasonic emulsification 1.5 hours, cross microporous filter membrane, spraying is dry, makes Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule.
Embodiment 7
Take the Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule 100g that embodiment 1～6 makes, add medicinal microcrystalline Cellulose 200g, mix homogeneously, wet granulation, water regulates, and makes loose particles and crosses 20 mesh sieves, dries.Filled capsules after dry.Obtain the capsule of Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule.
Embodiment 8
Take the Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule 100g that embodiment 1～6 makes, add 200g mannitol, after dissolving with aquesterilisa, cross 0.3 μ m microporous filter membrane, cannedly enter in ampere bottle,-50 ℃ of lyophilizations 24 hours, obtain the freezing-dried powder injection of Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule.
Embodiment 9
The particle diameter of the Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule that embodiment 1-6 makes and envelop rate experiment
Method: adopt second appendix IXE granularity of Chinese Pharmacopoeia in 2000 and particle size distribution method the 3rd method light scattering determining microcapsule diameter; Adopt acetone solution capsule material, filter, weigh the substance weight after volatilization acetone, envelop rate=(substance weight after the volatilization of nanocapsule weight-acetone)/nanocapsule weight.
Result: Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule diameter scope is between 200～2000nm, and envelop rate is 81～86%.The results are shown in Table one.This Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule diameter that shows that this employing embodiment 1～6 method makes is less, has higher envelop rate.
Table one Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule diameter distributes and envelop rate
Embodiment 10
The resisiting influenza virus experiment of the Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule that embodiment 1-6 makes
Method: 3 kinds of viruses (H3N2, H5N1, Influenza B virus) are diluted to respectively to 100 times of half viral infection concentration.The Cacumen Securinegae Suffruticosae resisiting influenza virus effective site water that embodiment 1～6 is made is mixed with 1% concentration, and press multiple and dilute, join the Testis et Pentis Canis passage cell (MDCK) that grows up to monolayer, every hole 50 μ l, the growth-promoting media that adds again influenza virus liquid 50 μ l/ holes and 100 μ l/ holes, every concentration 4 holes.Establish the medicinal liquid or the above-mentioned 3 kinds of viral contrasts that do not add by the preparation of Cacumen Securinegae Suffruticosae resisiting influenza virus effective site simultaneously.Put in 35 ℃, 5%CO2 incubator and spend the night, then in every hole, add 0.2ml 1% red cell suspension, mix homogeneously room temperature is placed 30～60min, observes blood clotting result and the toxicity of medicinal liquid to cell.
Result: medicinal liquid is 1: 4～16 to the limes null of cell, and the greatest dilution of anti-3 kinds of influenza virus is 1: 160～640, shows that Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule infected by influenza has stronger inhibition and killing action.The results are shown in Table two.
The greatest dilution of table two Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule resisiting influenza virus
Above-described embodiment is preferably embodiment of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under spirit of the present invention and principle, substitutes, combination, simplify; all should be equivalent substitute mode, within being included in protection scope of the present invention.
Claims (7)
1. a preparation method for Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule, is characterized in that, comprises the steps:
(1) to the ethanol that adds 5～10 times of its weight, mass fraction 30～80% in Cacumen Securinegae Suffruticosae, 30～80 ℃ of heating extraction 1～3 hour, filter, filtering residue repeats to extract 1～2 time with above-mentioned solvent again, filter, merging filtrate, in 30～60 ℃ of vacuum concentration to 1/3～2/3 of original volume, adds macroporous resin adsorption 8～24 hours; Take out macroporous resin, with mass fraction 50～90% ethanol or acetone eluting, 30～60 ℃ of vacuum concentration of eluent are to 1/3 of original volume～2/3 o'clock, with equal-volume dichloromethane, ethyl acetate or n-butanol extraction 1～5 time, combining extraction liquid, 30～60 ℃ of vacuum concentration are also dried 3～6 hours, obtain Cacumen Securinegae Suffruticosae resisiting influenza virus effective site; Described macroporous resin is D101, DM130, HPD600, AB-8, XAD-16 or NkA-9;
(2) capsule material is mixed with to the capsule material solution of 1～10% mass fraction with solvent, add the emulsifying agent of capsule material solution 0.5～5% mass fraction, the Cacumen Securinegae Suffruticosae resisiting influenza virus effective site of the antiplastering aid of 0.1～1% mass fraction, 1～20% mass fraction, ultrasonic emulsification 0.5～2 hour, cross microporous filter membrane, spraying is dry, makes Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule; Described solvent is one or more in methanol, ethanol, acetone, isopropyl alcohol, dichloromethane.
2. preparation method according to claim 1, it is characterized in that, the described capsule material of step (2) is one or more in acrylic resin, hydroxypropyl methylcellulose, ethyl cellulose, methylcellulose, gelatin, arabic gum, cyclodextrin, chitosan.
3. preparation method according to claim 1, is characterized in that, the described emulsifying agent of step (2) is non-ionic surface active agent.
4. preparation method according to claim 3, is characterized in that, described emulsifying agent is one or more in monoglyceride, Tween 80, sorbester p18, sucrose fatty acid ester and poloxamer.
5. preparation method according to claim 1, is characterized in that, the described antiplastering aid of step (2) is one or more in Polyethylene Glycol, Talcum, silica gel and magnesium stearate.
6. a Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule of preparing according to the preparation method of effective site microcapsule described in claim 1～5 any one.
7. the application of Cacumen Securinegae Suffruticosae resisiting influenza virus effective site microcapsule according to claim 6, is characterized in that, is made into resisiting influenza virus medicine or disinfectant anti-epidemic product.
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| CN108617882A (en) * | 2018-05-15 | 2018-10-09 | 广西万寿谷投资集团股份有限公司 | A kind of chicken feed addictive of anti-pestilence and preparation method thereof |
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| CN1230403A (en) * | 1998-12-22 | 1999-10-06 | 朱盛山 | Slow-releasing Chinese medicine capsule and its preparation |
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| CN1695647A (en) * | 2004-05-12 | 2005-11-16 | 四川省中药研究所 | Method for preparing general extractive from effective part of blattbulume |
| TW200607517A (en) * | 2004-08-17 | 2006-03-01 | Hepaguard Biotechnology Int L Co Ltd | Medicinal composition capable of suppressing proliferation or metastasis of tumor cells |
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