CN101371861A - High-efficient antiviral medicament composition in chickweed as well as preparation method and use thereof - Google Patents

High-efficient antiviral medicament composition in chickweed as well as preparation method and use thereof Download PDF

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CN101371861A
CN101371861A CNA2007101308293A CN200710130829A CN101371861A CN 101371861 A CN101371861 A CN 101371861A CN A2007101308293 A CNA2007101308293 A CN A2007101308293A CN 200710130829 A CN200710130829 A CN 200710130829A CN 101371861 A CN101371861 A CN 101371861A
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herba stellariae
stellariae mediae
compositions
stellaria
herba
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朱耕新
曾毅
李泽琳
钱裕盛
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Abstract

The invention discloses a highly effective antiviral medicinal composition of chickweed, a preparation method and an application thereof. Presently, highly effective, safe and universal antiviral medicine is lacking all over the world. A plant, chickweed, or other stellaria plant is extracted, by two resin adsorption methods, one water-alcohol extraction and ultra-filtration, into a dark brown composition that has a molecular distribution ranging from 2,000 to 900,000 Dolton and comprises total sulfate peptidoglycan phenolic acidic components and flavonoid components; the composition is used as a safer, more highly effective universal antiviral natural medicine. Total peptides account to 15-25 percent of the chemical structures of dark brown total sulfate peptidoglycan phenolic-acidic components, and include totaling 17 amino acids by proportion: aspartic acid, glutamic acid, serine, histidine, cystine, methionine, isoleucine; the polysaccharides essentially consist of glucose, galactose and arabinose and form sulfate polysaccharides. The composition can be used for treating virus diseases, including AIDS virus, hepatitis virus, respire virus such as influenza virus including highly pathogenic bird flu virus, para influenza virus and adenovirus, and papovavirus, enterovirus, mumps virus, herpes simplex virus, herpes zoster virus, and varicella-zoster virus, and on the like, does not show toxicity, and can be prepared into more than 10 medicinal dosage forms and healthcare products, and the production process does not cause pollution to the environment.

Description

A kind of high-efficient antiviral medicament composition and its production and application in the Herba stellariae mediae
One, technical field:
Publication number the inventor is on CN 1377680A and the CN 1498630A two patent of invention bases, this disclosure of the Invention progress and result that the compositions chemical analysis in above-mentioned two patents is furtherd investigate, relate to from the plant Herba stellariae mediae or from the molecular weight distribution of other a kind of Stellaria plant extract 2,000 dalton～900,000 dalton, be dark brown black total sulfuric ester peptidoglycan class the phenolic acid sexual element can with the compositions of Herba stellariae mediae flavonoid composition as a kind of safer, the broad-spectrum antiviral natural drug is used more efficiently.
Two, background technology:
By retrieval U.S.'s " chemical abstracts " (Chemical Abstracts) till the 134th volume, the chemical constituent of the Stellaria plant of being reported mainly is flavone and the former class of flavonoid glycosides, Saponin and Saponin, ring peptide class, ferulic acid and esters, chlorogenic acid, neochlorogenic acid, caffeic acid, quininic acid, ascorbic acid class, oxalic acid and calcium oxalate, monoacyl galactose lipoid, alkaloids, galactopyranose-SN-glycero class, chondrillasterol, stigmastenol, sitosterol class or the like from nineteen twenty to calendar year 2001.
The inventor was to disclose total organic phenolic acid and the glucoside compound thereof that contains more element sulphur and calcium constituent from Stellaria plant Herba stellariae mediae or other a kind of Stellaria plant in the molecular structure that extracts in the CN 1377680A patent and disclose the application of the high polarity compound compositions of the phenolic acid compound of sulfur-bearing in the structure as antiviral drugs in CN 1498630A patent at publication number once, and said composition and ferric chloride test solution show characteristics such as dark brown reaction.
Other has the patent of publication number CN1521193A to claim it to extract molecular weight distribution 4000～2000000 and the application of total flavones anti-herpesvirus and HIV (human immunodeficiency virus) in the Herba stellariae mediae that obtains with pure water lift alcohol deposition method that contained the canescence polysaccharide composition of peptide and carboxyl and patent disclosure that publication number is CN1521179A from the Stellaria plant with hot lower alcohol.
But do not see as yet so far from the plant Herba stellariae mediae or from the molecular weight distribution of other a kind of Stellaria plant extract 2, the report that the compositions of 000 dalton～900,000 dalton, the phenolic acid sexual element of total sulfuric ester peptidoglycan class that is dark brown black and flavonoid composition is used as broad-spectrum antiviral medicament.
Three, summary of the invention:
1, the technical problem to be solved in the present invention: provide a kind of from Herba stellariae mediae or the isolating molecular weight distribution of other a kind of Stellaria plant extract 2, the compositions of 000 dalton～900,000 dalton, the phenolic acid sexual element of total sulfuric ester peptidoglycan class that is dark brown black and flavonoid composition as safer, broad-spectrum antiviral medicament is used more efficiently.
2, the technical solution used in the present invention: on the disclosed CN 1377680A of the inventor and two parts of patent bases of CN1498630A, its compositions composition is further furtherd investigate, probe into its range of molecular weight distributions and chemical composition, adjust its ratio and make compositions have safer, broad-spectrum antiviral medicament effect more efficiently, thereby use as a kind of better antiviral drugs.
3, existing through on the gel column of high performance liquid chromatograph, using buffer solution elution, P-82 makes standard substance with glucosan, carrying out molecular weight distribution by GPC software records: from Herba stellariae mediae and other a kind of Stellaria plant extraction separation to the high-efficiency broad spectrum antiviral composition be that molecular weight distribution is 2,000 dalton～900,000 dalton, be the compositions that the flavonoid composition in phenolic acid sexual element and the Herba stellariae mediae of total sulfuric ester peptidoglycan class of dark brown black is formed in 55%～90% to 45%～10% ratio.
In the chemical constitution of the phenolic acid sexual element of this total sulfuric ester peptidoglycan class: its total peptide partly accounts for 15% to 25% and by comprising that following 17 seed amino acids are respectively by its composition of proportions: aspartic acid: 12.118%～13.18%, glutamic acid: 13.036%～18.425%, serine: 4.156%～6.19%, histidine: 1.89%～1.946%, twenty propylhomoserin: 8.909%～9.915%, threonine: 4.837%～7.129%, arginine: 2.92%～3.677%, alanine: 6.05%～9.647%, tyrosine 2.596%～2.877%, cystine: 1.89%～0.2%, valine: 5.94%～7.577%, methionine: 0.2%～2.09%, phenylalanine: 2.74%～3.738%, isoleucine: 4.28%～5.046%, leucine: 4.70%～5.53%, lysine: 5.77%～7.965%, proline: 4.61%～8.60%;
The content of element sulphur is 0.5～5% in the chemical constitution of the phenolic acid sexual element of this total sulfuric ester peptidoglycan class, except that institute's sulfur-bearings such as cystine, methionine, remaining sulfur and polysaccharide form polysaccharide sulfate or/and with sulfydryl or/and exist with the thioether form;
Sugar moieties in the chemical constitution of the phenolic acid sexual element of this total sulfuric ester peptidoglycan class mainly is the total polysaccharides that is made of glucose, galactose and arabinose, present the pale brown colour response of sugar and occurs two maximum absorption bands at 400nm～410nm and 480nm～490nm with phenol-strong sulfuric acid response, wherein the peak of 400nm～410nm is a main peak than about one times of the absorption peak height of 480nm～490nm appearance; Record its total polysaccharides through phenol-concentrated sulphuric acid method and in its chemical constitution, account for 55%～65%;
The phenolic acid sexual element of this total sulfuric ester peptidoglycan class and ferric chloride test solution manifest a kind of dark green dark brown reaction of phenols, and at 220nm～235nm place and 330nm～two maximum absorption bands of 350nm place appearance, wherein 220nm～235nm peak is a main peak, peak height than 330nm～350nm goes out one times, and the content of phenol is 10%～15% in its structure.
Compositions is soluble in water, and according to the different formation brownish reds of the phenolic acid sexual element of total sulfuric ester peptidoglycan class wherein and flavonoid component content and concentration, dark brown red or near brownish black clear solution, molecular weight is big more in the phenolic acid sexual element of this total sulfuric ester peptidoglycan class, content is high more, and low more its solution color and luster of the content of flavone tends to the still extremely near black of nearly dark brown black more, its antivirus action is also strong more.
4, compositions of the present invention, be from a following kind of plant, to extract, these plants have comprised Stellaria plant Stellaria L. and cattle Stellaria plant Malachium Fries following 79 kinds, i.e. Herba stellariae mediae Stellariamedia (L.) Cyr., Stellaria alsine Grim. Stellaria alsine Grimm., brown lobe Stellaria alsine Grim. Stellaria alsinevar.phaeuspetala Hand.-Mazz., Anhui Herba stellariae mediae Stellaria anhwiensis Migo., blunt calyx Herba stellariae mediae Stellaria amblyosepala Schrenk., apicule Herba stellariae mediae Stellaria apiculata Wils., Herba malachii aquatici Stellariaaquatica (L) Scop., the husky Herba stellariae mediae Stellaria arenaria Maxim that gives birth to, Alishan Herba stellariae mediae Stellariaarisanensis Hayata., slender lobule Alishan Herba stellariae mediae Stellaria arisanensis var.leptophylla Hayata., north Herba stellariae mediae Stellaria borealis Bigel., short lobe Herba stellariae mediae Stellaria brachypetala Bge., long lobe Herba stellariae mediae Stellaria bungeana Fenzl., northeast Herba stellariae mediae Stellaria cherleriae (Fisch.) Will., China Herba stellariae mediae Stellaria chinensis Regel, Taiwan Herba stellariae mediae Stellaria cicrantha Hayata, thick leaf Herba stellariae mediae Stellariacrassifolia Ehrh., wrinkle leaf Herba stellariae mediae Stellaria crispate Wall., David's Herba stellariae mediae Stellaria davidiiHemsl., the Herba stellariae mediae Stellaria decumbens Edgew. that crouches lays down, the needle-like Herba stellariae mediae Stellaria decunbensvar.acicularia Edgew.et Hook.f. that crouches that lays down, southwest Herba stellariae mediae Stellaria delavayi Franch., Stellaria dianthifolia Williams Stellaria dianthifolia Williams, forked cyme Herba stellariae mediae Stellaria dichasioides Williams, bifid Herba stellariae mediae Stellaria dichotoma L., narrow leaf bifid Herba stellariae mediae Stellaria dichotoma var.lanceolataBge., line leaf bifid Herba stellariae mediae Stellaria dichotoma var.stepheniana Willd., Herba stellariae mediae Stellariadiffusa Wills. looses in the shop, turn over white Herba stellariae mediae Stellaria discolor Turcz., Stellaria diversiflora Maxim. Stellaria diversifloraMaxim., the different colored Herba stellariae mediae Stellaria diversiflora var.gymnandra Franch. of naked stamen, concave veins Herba stellariae mediae Stellaria depressa Schnid., Du Shi Herba stellariae mediae Stellaria duthiei Gandoger, line stem Herba stellariae mediae Stellariafilicaulis Mak., line handle Herba stellariae mediae Stellaria filipes Komar., spend more Herba stellariae mediae Stellaria florida Fisch., standing grain leaf Herba stellariae mediae Stellaria graminea L., dredge pubescence standing grain leaf Herba stellariae mediae Stellaria graminea var.pilosulaMaxim., turn green standing grain leaf Herba stellariae mediae Stellaria graminea var.viridescens Maxim., Jiangzi's Herba stellariae mediae Stellaria gyantsensis Williams, XIACAO Herba stellariae mediae Stellaria gypsophiloides Fenzl., Stellaria henryi williams Stellaria henryi Williams, the Herba stellariae mediae Stellaria hsinganensis Kitagawa of Xingan, introversion Herba stellariae mediae Stellaria infracta Maxim., slender lobule Herba stellariae mediae Stellaria leptophylla Hance, Stellaria maximowixziana Franch. Stellariamaximowixziana Franch., Herba stellariae mediae Stellaria media (L.) Cyr., Stellaria micrantha Hayata Stellariamicrantha Hayata, gentle Herba stellariae mediae Stellaria mitans Williams, goose intestinal Herba stellariae mediae Stellaria neglectaWeihe, Herba stellariae mediae Stellaria neo-palustris Kitagawa is given birth in new natural pond, eight stamen Herba stellariae mediae Stellaria octandraFobedim., raspberry Herba stellariae mediae Stellaria oxycoccoides Komar., Herba stellariae mediae Stellaria palustris L. is given birth in the natural pond, exhibition leaf Herba stellariae mediae Stellaria patentifolia Kitagawa, handle flower Herba stellariae mediae Stellaria peduncularis Bge., handle leaf Herba stellariae mediae Stellaria petiolaris Hand-Mazz., Herba stellariae mediae Stellaria pilosa Franch. becomes mildewed, imitation stone is given birth to Herba stellariae mediae Stellaria pseudosaxatilis Hand.-Mass., blinks Stellaria pusilla Schmid., flint lobe Herba stellariae mediae Stellaria radians L., net arteries and veins Herba stellariae mediae Stellaria reticulivena Hayata, rock Herba stellariae mediae Stellariarupestris Hemsl., stone is given birth to Herba stellariae mediae Stellaria saxatilis Buch.-Ham., embrace stem stone and give birth to Herba stellariae mediae Stellariasaxatilis var.amplexicaulis Hand.-Mazz., accurate Ge Er Herba stellariae mediae Stellaria soongorica Roshev., Su Shi Herba stellariae mediae Stellaria souliei Williams, star hair Herba stellariae mediae Stellaria stellato-pilosa Hayata, garden calyx Herba stellariae mediae Stellaria strongylosepala Hand.-Mazz., intend umbrella flower Herba stellariae mediae Stellaria subumbellataEdgew., little fine hair Herba stellariae mediae Stellaria tomentella Ohwi, three type Herba stellariae mediae Stellaria trimorpha Nakai, Turkestan Herba stellariae mediae Stellaria turkestanica Schischk., wetland Herba stellariae mediae Stellaria uda Williams, umbrella flower Herba stellariae mediae Stellaria umbellata Turcz., LVHUA Herba stellariae mediae Stellaria virdiflora Pax et O.Hoffm., Stellaria wushanensis Williams Stell aria wushanensis Williams, five Herba stellariae mediae Stellaria wutaicaHand.-Mazz., Yunnan Herba stellariae mediae Stellaria yunnanensis franch.j or cattle Stellaria plant cattle Herba stellariae mediae Malachium aquaticum (L.) Fries etc.
5, the present invention also provides with form sediment again the extraction through ultrafiltration of water extraction precipitate with ethanol and prepares method for compositions and heat and improve the method that improves the compositions yield on the resin column behind the elution requirement, and with the lower alcohols eluting compositions of low concentration with the phenolic acid sexual element content that improves total sulfuric ester peptidoglycan class in the compositions, reduce the wherein content of flavonoid composition, the antivirus action of enhancing composition reduces toxic and side effects and zest and production cost.
6, compositions has following beneficial effect:
But the compositions antiviral comprises: HIV (human immunodeficiency virus), hepatitis virus, the influenza virus that comprises high pathogenic avian influenza virus, parainfluenza virus, Respirovirus and papovaviruses such as adenovirus, enterovirus, mumps virus, herpes simplex virus, varicella zoster virus and varicella-zoster virus etc., and than having the phenolic acid sexual element of more high-load total sulfuric ester peptidoglycan class and reduce the flavonoid component content in the inventive compositions in the disclosed CN 1377680A of the inventor and two parts of patents of CN 1498630A, thereby compositions has stronger antivirus action and lower toxic and side effects or zest.
Four, the specific embodiment:
One), the preparation of compositions method comprises following three kinds:
1, its preparation method 1, its step comprises: crude drug or the bright grass of plant Herba stellariae mediae or other a kind of Stellaria plant are converted to Radix Glycyrrhizae, pulverize the back and obtain aqueous solution through precipitation with water extraction, centrifugal or filter the back by the macroporous resin column handled well in advance to adsorb the composition of described compositions, wash macroporous resin column, again macroporous resin column is heated and after 45 ℃～55 ℃, wash post and collect effusive dark brown distilled water liquid with the thermal distillation of same temperature, continue with heating to 45 ℃～55 ℃, concentration is at 20%～35% lower alcohols or acetone eluting from the resin column, collect brownish red or dark reddish brown tryptophol liquid or acetone solution that eluting obtains, to merge with the dark brown water liquid of collecting behind this solution recovery alcohol or the acetone, be concentrated into than weighing and left standstill 12 hours after 0.80 to 1.00, centrifugal removal impurity, promptly get the compositions crude product through 60 ℃ of vacuum dryings or spray drying, the phenolic acid sexual element of total sulfuric ester peptidoglycan class and the ratio of flavonoid composition are 55%:45% in the compositions crude product; As adding concentration behind the solution of compositions crude product reuse pure water being dissolved into proportion 0.80～1.10 is that 95% ethanol makes in the solution alcohol or acetone concentration reach 50%～70%, leave standstill, the leaching precipitate and with 95% washing with alcohol of 3 times of volumes once, the washing with acetone of 2 times of volumes once, through 60 ℃ of vacuum dryings or be dissolved in water into proportion be 1.00～1.20 solution again spray drying promptly get compositions, the phenolic acid sexual element of total sulfuric ester peptidoglycan class and the ratio of flavonoid composition are 90%:10% in the said composition; Wherein lower alcohols comprises methanol, ethanol, propanol and isopropyl alcohol.
2, the described preparation of compositions method 2 of claim 1, its step comprises: the back is cleaned, pulverized to the bright grass of plant Herba stellariae mediae or other a kind of Stellaria plant or bright Radix Glycyrrhizae extract aqueous solution, with this aqueous solution through precipitation, centrifugal or filter after obtain liquid; This liquid is adsorbed on described compositions composition on the post through the anionite post, with the 2N sodium chloride solution described compositions composition is eluted from post, again through desalting processing, be concentrated into than weighing and left standstill 12 hours after 0.80 to 1.00, centrifugal removal impurity, add concentration and be 95% ethanol and make that concentration of alcohol reaches 50%～70% in the solution, leave standstill, leaching precipitate and with 95% washing with alcohol of 3 times of volumes 1～2 time, the washing with acetone of 2 times of volumes 1～2 time is through 60 ℃ of vacuum dryings or to be dissolved in water into proportion be that 60 ℃ of vacuum dryings or spray drying promptly get compositions again for 1.10～1.20 solution;
Wherein the anionite post comprises weak anion exchange resin post and cellulose anion exchanger post.
3, the described preparation of compositions method 3 of claim 1, its step comprises: after the dried pulverizing medicinal materials with plant Herba stellariae mediae or other a kind of Stellaria plant, with twice of the flooding of 5～10 times of volumes, each 1～3 hour, the dark-brown extracting solution that united extraction obtains, be evaporated to proportion 0.7～0.9, left standstill 12 hours, the filtering precipitation, adding 95% lower alcohols or acetone make concentration reach 60%～70% in the solution, left standstill 12 hours, the precipitate that the filter collection is separated out promptly gets the compositions crude product, this compositions crude product is dissolved after high speed centrifuge 15 with an amount of pure water, 000rpm, centrifugal 15 minutes, clear liquor elder generation via hole diameter after centrifugal is that the ceramic membrane treatment facility fine straining of 0.22 μ m is handled, be ultrafilter membrane ultrafiltration more than 2000 through molecular weight cut off again, will be through the solution concentration after part small-molecular weight impurity is removed in ultrafiltration to proportion 0.7～0.9, adding lower alcohol or acetone makes concentration reach 85% to carry out the secondary precipitate with ethanol, leave standstill the sucking-off supernatant after 12 hours, precipitate is closely dried through decompress filter, in time successively add an amount of dehydrated alcohol and washing with acetone, sucking filtration is extremely done, and promptly gets purer powdered composition through 60 ℃ of vacuum dryings immediately; Wherein lower alcohol is methanol, ethanol, propanol, isopropyl alcohol.
4, compositions is used as antiviral drugs and can be made following dosage form at least:
1). injection can be used for treating atypical pneumonia, the high pathogenic avian influenza virus H of acquired immune deficiency syndrome (AIDS), hepatitis, the initiation of SAS virus 5N 1The various viral diseases that type etc. cause.
2). injectable powder can be used for treating atypical pneumonia, the high pathogenic avian influenza virus H of acquired immune deficiency syndrome (AIDS), hepatitis, the initiation of SAS virus 5N 1The various viral diseases that type etc. cause.
3). aerosol is used for children's and treats influenza, parotitis etc., and can be used as the antiviral disinfectant of air and oral cavity, health product or antiviral functional food.
4). buccal tablet or chewable tablet, be used for the treatment of parotitis etc., can be used as the antiviral disinfectant in air and oral cavity, health product or antiviral functional food.
5). capsule, the oral various viral diseases such as acquired immune deficiency syndrome (AIDS), hepatitis, influenza that are used for the treatment of.
6). drop pill, micropill, microcapsule etc., in order to treatment acquired immune deficiency syndrome (AIDS), hepatitis chronic diseases poison disease.
7). nasal drop or nasal spray, be used for the treatment of influenza etc., also can be used as the antiviral disinfectant of nasal cavity, health product etc., also can be by the nebulizer inhalation that atomizes, in order to treatment treatment viral pneumonia, as atypical pneumonia, the high pathogenic avian influenza virus H of SAS virus initiation 5N 1The viral disease that type causes etc.
8). membrane or varnish etc. are used for treating for skin disease such as herpes, condyloma acuminatum.
9). mucilage is used for the treatment of dermatosiss such as herpes.
10). water preparation is used for the clinical dermatosiss such as treatment herpes, condyloma acuminatum of directly smearing.
11). the liquid chewing gum can be used as oral cavity antiviral disinfectant, antiviral health product or functional food.
12). chewing gum can be used as oral cavity antiviral disinfectant, antiviral health product or functional food etc.
Two), the embodiment of compositions:
1, with the embodiment 1 of first method: get the bright careless 32kg conversion of the Herba stellariae mediae of choosing clean impurity and claim to add 8～12 times water extraction behind the Radix Glycyrrhizae, centrifugal, filtration, extract medicinal liquid, with the commercially available macroporous resin column of this medicinal liquid by id30cm * 120cm, the control flow velocity is at 5ml/min.As seen macroporous resin column from top to bottom becomes a kind of brownish red or dark brown brown is filled medicine until pillar gradually.Discard effusive liquid in the pillar, and earlier with the residual liquid in the clean post of tap water.With 55 ℃ of distilled water wash macroporous resin column of about 2 times of column volumes, discard effusive at first colourless and water liquid that color and luster is more light, collect effusive subsequently dark brown water liquid.This moment, macroporous resin column was heated to 45 ℃～50 ℃, and continuing is about 35% ethanol elution of 3 times of amounts of column volume and collects the brownish red alcoholic solution with 55 ℃, and this routine consumption is about the ethanol of 300L 35%; The brownish red alcoholic solution of collecting is reclaimed ethanol through concentrating under reduced pressure, the gained concentrated solution is concentrated into proportion 0.80 with the dark brown water liquid merging of collecting, be about about 1010ml, standing over night, centrifugal 5 minutes then with 4000r/min, discard precipitate, supernatant through 0.5 μ m membrane filtration in sterilized clean container, measure wherein half after being concentrated into proportion 1.10, in 60 ℃ of vacuum dryings, and the ratio of the phenolic acid sexual element that records wherein total sulfuric ester peptidoglycan class and flavone is 55%:45%; All the other half solution become proportion through concentrating under reduced pressure again under aseptic condition be 1.00 thick paste shape, left standstill again 12 hours, centrifugal removal impurity, add concentration and be 95% ethanol and make that determining alcohol reaches 70% in the solution, leave standstill, the leaching precipitate and with 95% washing with alcohol of 3 times of volumes once, the washing with acetone of 2 times of volumes once, promptly get compositions through 60 ℃ of vacuum dryings, and the ratio that records the wherein phenolic acid sexual element of sulfuric ester peptidoglycan class always and flavone is 90%:10%;
Compositions is dark brown brown loose powder, and the common abnormal smells from the patient of Chinese medicine is arranged, count 70.38g, yield is calculated as 2.20 ‰ by bright grass, makes compositions 606g altogether with this method repeatedly.
2, with the embodiment 2 of second method: get the dried herb 5kg of fresh Herba stellariae mediae, pulverize after cleaning earth with tap water, use the water extraction medicinal liquid; With medicinal liquid through precipitation, centrifugal or filter after promptly obtain liquid, with the commercially available anion-exchange resin column of this liquid by id 10cm * 120cm, after adsorbing described compositions composition, clean the weak-base anion-exchange resin post with water recently distilled that is about 3 times of column volumes or deionized water, discard from post effusively by light yellow thin out gradually until the almost colourless post liquid of washing, this routine consumption is about 30L water; Then resin column is heated to 45 ℃～50 ℃ continue with 2 times to column volume and to be heated to about 55 ℃ concentration be that the NaCl solution of 2N will be adsorbed on the rapid eluting of described compositions composition on the post and collect this dark brown red eluent, this example is 20L with the NaCl solution amount.This saline solns suitably is concentrated into is placed on about 750ml in the semipermeable membrane bag, successively tap water and distilled water are dialysed until solution to AgNO 3Solution reaction is very weak or do not have till the saline taste; The back gained dark brown red solution of will dialysing is transferred to that drying under reduced pressure to proportion 0.90 is thick in the vacuum drying apparatus, is about 400ml, and standing over night discarded precipitate in centrifugal 5 minutes with 4000r/min then; Add concentration and be 95% ethanol and make that concentration of alcohol reaches 70% in the solution, leave standstill, leaching precipitate and with the washing with acetone of 1～2 time, the 2 times volume of 95% washing with alcohol of 3 times of volumes 1～2 time, get dark brown brown powder through 60 ℃ of vacuum dryings, general Chinese medicine abnormal smells from the patient is arranged, altogether 75.65g, yield is calculated by Radix Glycyrrhizae and is about 1.513%, makes highly finished product 512.73g altogether with this method.
3, with the embodiment 3 of the third method: after getting the bright Radix Glycyrrhizae 5kg of plant Herba stellariae mediae and cleaning, pulverize, water 15kg, 15kg extract twice, merge obtained aqueous solution, with this aqueous solution through precipitation, centrifugal or filter after obtain liquid; Is about 0.8 with this liquid through being evaporated to proportion, add 95% ethanol, make concentration of ethanol reach 75%, a large amount of muddinesses promptly appear in solution, leave standstill and spent the night in 12 hours, the sucking-off supernatant discards, with the lower sediment thing through centrifugal or filter the compositions crude product, this crude product reuse 400ml pure water is dissolved after high speed centrifuge 15,000r/min, centrifugal 15 minutes is the ceramic membrane treatment facility fine straining processing of 0.22 μ m with the elder generation of the clear liquor after centrifugal via hole diameter, be ultrafilter membrane ultrafiltration more than 2000 through molecular weight cut off again, will be through the solution concentration after part small-molecular weight impurity is removed in ultrafiltration to proportion 0.85, add 95% ethanol and make and reach 85% concentration and carry out the secondary precipitate with ethanol, leave standstill the sucking-off supernatant about 12 hours, precipitate is closely dried through decompress filter, in time successively add dehydrated alcohol 350ml and acetone 350ml washing, sucking filtration is extremely done, and promptly gets purer dark brown brown powder shape compositions 82.29g through 60 ℃ of vacuum dryings immediately, yield is 1.65%, makes 465g altogether with this method.
When table 1. provided on macroporous resin column the eluting compositions, variations in temperature was to the influence of yield:
Among table 1. embodiment 1 from post during the eluting compositions variations in temperature to the influence of yield
Figure A200710130829D00111
Figure A200710130829D00121
Concentration of alcohol changed the influence to yield when table 2. was given on macroporous resin column eluting compositions:
Among table 2. embodiment 1 from post during the eluting compositions concentration of alcohol change influence to yield
Figure A200710130829D00122
4, molecular weight distribution determination: get and extract resulting composition 2g and after water alcohol precipitation 3 times is colourless to pure liquid, can think again and the flavonoid composition in the compositions is cleaned as for test agent (calling " supplying test agent " in the following text), will be for the test agent water dissolution, sample introduction on high performance liquid chromatograph, produce row's group with Japanese Shodex and be limited to one megadalton OHpak SB-804 HQ post, through 0.1% sodium phosphate buffer eluting, flow velocity: 0.6ml/min, measure wavelength: 220nm, produce glucosan STANDARD P-82 with Japanese Shodex and make standard substance, carrying out molecular weight distribution determination by GPC software gets: from Herba stellariae mediae extraction separation to its molecular weight distribution of phenolic acid sexual element of total sulfuric ester peptidoglycan class 2,000 dalton～900,000 dalton.
5, amino acid content is measured in the composition sample: get composition sample (containing the flavonoid composition) 0.2g and add 6N HCl in 110 ℃～135 ℃ tube sealing hydrolysis 20～21 hours, neutralization back standardize solution is got the supernatant standard amino acid and is to contrast and record amino acid content and be: aspartic acid: 12.118%, glutamic acid: 13.036%, serine: 4.156%, histidine: 1.89%～, glycine: 8.909%, threonine: 4.837%, arginine: 2.92%, alanine: 6.05%, tyrosine 2.596%, cystine: 1.89%, valine: 5.94%, methionine: 0.2%, phenylalanine: 2.74%, isoleucine: 4.28%, leucine: 4.70%, lysine: 5.77%, proline: 4.61%; And to record total amino acids content be 15%.
That gets above-mentioned clean flavone again adds 6N HCl in 110 ℃～135 ℃ tube sealing hydrolysis 20～21 hours for test agent 0.2g, neutralization back standardize solution is got the supernatant standard amino acid and is to contrast and record amino acid content and be: aspartic acid: 13.18%, glutamic acid: 18.425%, serine: 6.19%, histidine: 1.946%, glycine: 9.915%, threonine: 7.129%, arginine 3.677%, alanine: 9.647%, tyrosine 2.877%, cystine: 0.2%, valine: 7.577%, methionine: 2.09%, phenylalanine: 3.738%, isoleucine: 5.046%, leucine: 5.53%, lysine: 7.965%, proline: 8.60%; And to record total amino acids content be 25%.
Simultaneously, this has detected glucose, galactose and arabinose in the acid hydrolysis liquid of test agent.
6, phenol chromogenic reaction and content measuring for test agent: take a morsel and dissolve with pure water for test agent, dripping 2 2N HCl makes and is acid, drip two of 1% ferric chloride test solutions and promptly manifest a kind of dark green dark brown reaction of phenols, through the scanning of day island proper Tianjin product UV-2401PC ultraviolet-visible photothermal spectroscopic analyzer, sweep limits: 200nm-900nm, as seen at 220nm～235nm place and 330nm～350nm place two maximum absorption bands appear, wherein 220nm～235nm peak is a main peak, peak height than 330nm～350nm goes out one times, and to record phenol content at the 343.50nm place be 10%～15%.
7, supply the phenol-concentrated sulphuric acid chromogenic reaction and the test of total polysaccharides in the test agent: take a morsel and dissolve with pure water 1ml for test agent, drip 2 freshly prepared 5% phenol reagent, pour the pale brown color that the 5ml concentrated sulphuric acid promptly presents sugar rapidly, 15min in boiling water again, through the scanning of day island proper Tianjin product UV-2401PC ultraviolet-visible photothermal spectroscopic analyzer, sweep limits: 200nm-900nm, as seen two maximum absorption bands occur at 400nm～410nm and 480nm～490nm, wherein the peak of the 400nm～410nm absorption peak height that to be main peak occur than 480nm～490nm is about one times; Through be contrast with the glucose standard substance, to record total polysaccharides content at the 485nm place be 55%～65%.
8, for the sulfuric ester spectroscopic test in the test agent: get for test agent and use the KBr tabletting on a small quantity, on infrared scanner at 4000cm -1～400cm -1Wave-number range scans, visible 1236.94cm in its infrared scan collection of illustrative plates -1The wave number peak, showing has OSO 3The S=O stretching vibration, 838.14cm -1The wave number peak show the stretching vibration that C-O-S is arranged, this is the characteristic peak of thioester bond, shows in the chemical constitution of phenolic acid sexual element of this total sulfuric ester peptidoglycan class to have the sulfuric ester structure; Wherein O-S is that sulfide based structural, S=O are sulfhydryl structures.
9, the preparation embodiment of various dosage forms is as follows:
9.1. injection: the medicine highly finished product 200g that the embodiment extraction separation is obtained dissolves back centrifugal 5 minutes of centrifuge (4000r/min) with 0.9% sodium chloride injection 2000ml, the gained supernatant of centrifugal back is filtered in sterilized clean container through the sealing of 0.5 μ m filter membrane, use in the ampoule in injection through the filling and sealing machine fill, be distributed into respectively optionally that 1ml/ props up 1000,0.1g/ prop up, 450 of propping up of 2ml/, 0.2g/ prop up, and promptly get injection through cobalt 60 radiation sterilizations again.
9.2. injectable powder: the medicine highly finished product 200g that the embodiment extraction separation is obtained dissolves after centrifugal 5 minutes of centrifuge (4000r/min) with distilled water 50ml, the gained supernatant of centrifugal back is filtered in sterilized clean container through the sealing of 0.5 μ m filter membrane, the spray of spray-dried machine is dried to become medicated powder, be potted in the ampoule through filling and sealing machine again, promptly can be made into the injectable powder of 2ml again through cobalt 60 radiation sterilizations, amount to 2000,100mg/ props up.Use with sterilized water for injection dissolving back during use.
9.3. aerosol: get 4g medicine highly finished product add 0.9% sodium chloride injection 100ml dissolving, centrifugal, filter in the 10ml of cleaning dress aerosol bottle, promptly make 10 aerosols, 200mg/ props up.Can be used as the antiviral disinfectant in oral cavity.
9.4. contain tablet: the dressing, spice, correctives etc. one of getting the medicine highly finished product 100g that makes by embodiment 1 and tablet is reinstated tablet machine and is made 2000 of buccal tablets, the 50mg/ sheet.Can be used as the antiviral disinfectant in oral cavity, health food or antiviral functional food.
9.5. capsule: be ground into powdered highly finished product 100g under aseptic condition after getting the medicine vacuum drying that makes by embodiment 1, fill becomes 1000 capsules, the 100mg/ capsule.Can be used for treating various virosiss, also can be used as the antiviral disinfectant in oral cavity, health food or antiviral functional food.
9.6. microcapsule: the medicine dry powder 3g that gets the process refinement treatment that makes by embodiment 1 makes and is suspended in the 3.6g liquid Paraffin; Get I hundred glue 10g more in addition and add distilled water 200ml, boil half an hour with disruptive oxidation enzyme after waiting to dissolve, warm liquid Paraffin is injected my hundred glues, emulsifying is 1～2 minute in tissue mashing machine, makes oil in water emulsion.Get gelatin 10g again and add distilled water 200ml, place in 60 ℃ of water-baths and dissolve, and be mixed in the 1000ml beaker, keep glue, stir at a slow speed at 45 ℃～50 ℃ with above-mentioned my hundred glues.Add in the glue with freshly prepared 5% acetum 5ml, the pH value of regulator solution produces cohesion to being about 4.1.Inject distilled water 750ml behind the encystation, move in the ice bath again and cool off, be cooled to 5～10 ℃, the gelatin cyst membrane is congealed.At this moment microcapsule is spherical.Adding 37% formaldehyde 7ml again is cured.Stirred 15 minutes, and made the microcapsule typing.Regulate capsule liquid to the PH8.0 with 20% sodium hydroxide solution then.After low temperature stirs 1 hour, filter with centrifuge, washing is to neutral formaldehydeless flavor.Add capsule again and weigh 3% magnesium stearate and make diluent, cross 16 mesh sieves behind the mix homogeneously, 50 ℃ of vacuum dryings again, the microcapsule of this medicine, yield 72.2%, meter 16.6g microcapsule, 180mg/g.
Also can be made into micropill, drop pill etc. in addition.
9.7. nasal drop: will dissolve back centrifugal 5 minutes of centrifuge (4000r/min) with 0.9% sodium chloride injection 100ml by the medicine highly finished product 2g that embodiment 1 extraction separation obtains, the gained supernatant of centrifugal back filtered in sterilized clean plastics eyedrops bottle through the sealing of 0.5 μ m filter membrane be distributed into 10 nasal drop, 200mg/ props up.
9.8. membrane: get the medicine highly finished product 30g, the azone 0.3ml that make by embodiment 1 add water 100ml dissolving back centrifugal, through 0.5 μ m membrane filtration in clean container, add the suitable heated and stirred of PVA (05-88) 19.5g and make its dissolving, on plate glass, paint wide 100mm under the cleaning condition, be about the medicine film of 100mm, thick about 0.1mm.Lucifuge coating film with wide about 100mm, long 100mm, thick about 0.05mm respectively covers one deck as coating membrane at medicine film upper and lower surface again, is packaged into membrane then.Its dosage is 0.3g/cm 2, face the time spent on demand and dosage shear to use.
9.9. mucilage: get Pseudobulbus Bletillae (Rhizoma Bletillae) colloid (medium powder) 2g and be sub-packed in the dry glass bottle of 10 15ml, each adds glycerol 2ml and makes dispersant, after jolting is uniformly dispersed, other gets the stirring of chloroform water 100ml adding 3g medicine highly finished product and makes its dissolving, centrifugal after 0.5 μ m filter membrane filters respectively in finely dispersed Pseudobulbus Bletillae (Rhizoma Bletillae) Plexiglas's bottle, fierce jolting gets the medicine mucilage, the 200mg/ bottle.
9.10. water preparation or nasal drop: the medicine dry powder 10g that gets the refinement treatment that makes by embodiment 1 makes and is dissolved in the 250ml water, can be distributed into 25 bottles of water preparations, the 400mg/ bottle.
9.11. varnish: the dry extract powder of this product can be made into varnish.
9.12, membrane: get the medicine highly finished product 30g, the azone 0.3ml that make by embodiment 1 add water 100ml dissolving back centrifugal, through 0.22 μ m membrane filtration in clean container, add the suitable heated and stirred of PVA (05-88) 19.5g and make its dissolving, on plate glass, paint wide 100mm under the cleaning condition, be about the medicine film of 100mm, thick about 0.1mm.Lucifuge coating film with wide about 100mm, long 100mm, thick about 0.05mm respectively covers one deck as coating membrane at medicine film upper and lower surface again, is packaged into membrane then.Its dosage is 0.3g/cm 2, face the time spent on demand and dosage shear to use.
9.13, the liquid chewing gum: in water preparation, add spice, correctives etc. and make the liquid chewing gum, can be used as oral cavity antiviral disinfectant, antiviral health food or functional food etc.
9.14. chewing gum: 100g chewing gum glue, 0.1ml Oleum menthae spice, pentose 1g make chewing gum as correctives etc. and can be used as oral cavity antiviral disinfectant, antiviral health food or functional food etc.
10, compositions is as the safety of medicinal application
10.1, the acute toxicity testing of mice oral administration: to be the phenolic acid sexual element of total sulfuric ester peptidoglycan class and Herba stellariae mediae flavonoid form (down with) by 7:3 to compositions, after observation once or is for several times given mice compositions saturated solution, acute toxic reaction that experimental animal produced and death condition are for clinical consumption provides reference frame.
10.1.2, test material
Animal: healthy regular grade Kunming mouse, body weight 18～22g, male and female half and half are provided by Anhui Prov. Medical Science Inst.'s laboratory animal laboratory.The quality certification: No. 01, the real moving accurate word of Anhui doctor.
Medicine: composition powder (the same); Be made into desired concn with distilled water before using.
10.1.3, test method:
The saturated solution of composition powder (400mg/ml): get 12 of white mice, fasting 14～16 hours, body weight 19-21 gram, male and female half and half.Look sex and body weight animal is divided into 3 groups at random, 4 every group, the agent distance is than 1:0.5 between group.Each treated animal is irritated the solution (400mg/ml, 200mg/ml, 100mg/ml) that stomach gives composition powder with the 0.4ml/10g body weight respectively, observes oral being subjected in one week of reagent, causes the fatal dose of laboratory animal 0% and 100% death and the symptom that animal is showed.Result: after 3 treated animals are taken medicine, all find no animal poisoning symptom and death condition take place, so adopt the maximum dosage-feeding test.
Maximum dosage-feeding test (MTD):
Composition powder: get 20 of mices, male and female half and half, body weight 18～20g.In 24 hours, be administered twice (twice dosing interval is 6 hours) for respectively each Mus continuous irrigation stomach, irritate stomach at every turn and give composition powder saturated solution (400mg/ml) 0.8ml/20g, accumulative total gavages 1.6ml, instant rising observed mice poisoning and death condition, observes continuously 7 days.
10.1.4, result of the test:
(1). test mice body weight change situation:
In the week, all do not have the death of appearance behind the sample sets animals administer, the activity of animal, the mental status, diet, defecation etc. there is no unusually.The mice body weight change sees Table 3. respectively when testing preceding and off-test:
Table 3. composition powder acute toxicity test mice body weight change (X ± S)
Figure A200710130829D00161
(2). the maximum dosage-feeding of composition powder calculates:
The mice maximum tolerated dose: the compositions saturated solution contains 400 milligrams of composition powders for every milliliter, and the mice average weight is 20g, irritates stomach volume 0.8ml at every turn, amounts to 2 times.Mice day oral compositions powder maximum tolerated dose=1.6ml * 0.4g/ml/0.02kg=32g/kg
Margin of safety: 60kg is a standard with adult's average weight, and the consumption per day of composition powder is 1g; The mice average weight is 20g, and a day maximum dosage is 1.6ml * 0.4g/ml, margin of safety=0.64g/20g ÷ 1g/60000g=1916.17 (doubly)
10.1.5, conclusion:
Above-mentioned data show that acute toxicity test does not see that overt toxicity etc. is arranged in the animal body of composition oral administration, so compositions is a kind of very safe natural drug.
Compositions mice oral administration, maximum tolerated dose is 32g/kg, than the inventor once in CN1377680A and two parts of patents of CN 1498630A disclosed oral administration maximum tolerated dose be better about 3 times of 11.25g/kg, be equivalent to 1916.17 times of clinical administration daily dose.The experimental result explanation: compositions does not produce the overt toxicity reaction under animal oral administration condition, sample sets laboratory animal body weight natural increase during the laboratory observation, activity, the mental status, diet and defecation etc. are not seen obvious change yet, showing that compositions toxicity is lower, is safe by drafting that clinical dosage takes.
11, experiment of the general pharmacology of compositions and result thereof:
11.1, to the neural influence of spirit of mice and rat with to the influence of the sleeping number of mice:
Experiment conclusion:
(1) compositions nasal drop 0.67g/kg, 0.33g/kg, 0.16g/kg three dosage groups to mice pentobarbital sodium threshold down sleep dosage do not have synergism, relatively do not have significant difference (P〉0.05) with the pentobarbital sodium group, prompting compositions nasal drop does not have syngignoscism to mice.
(2) compositions nasal drop 0.67g/kg, 0.33g/kg, 0.16g/kg different time sections is to the spontaneous activity of mice counting after administration for each dosage group, with matched group there was no significant difference (P〉0.05) relatively, prompting compositions nasal drop does not have influence to the central nervous system of mice.
(3) compositions nasal drop 0.33g/kg, 0.16g/kg, each dosage group of 0.08g/kg different time sections rat transfer rod after the administration number of times that drops does not relatively have significant difference (P〉0.05) with matched group, and prompting compositions nasal drop is to the not influence of behavior of rat.
In sum, the compositions nasal drop is to the movable not influence of the normal behaviour of rat, mice, with threshold down the pentobarbital sodium of sleep dosage do not have synergism, showing that the compositions nasal drop does not have obviously the spiritual nervous system of animal in being subjected to the amount of reagent scope influences.
11.2, compositions is to the influence of anesthetized dog cardiovascular and respiratory system
11.3, experiment conclusion: compositions 0.67g/kg, 0.33g/kg, the basic, normal, high dosage group of 0.16g/kg does not have obviously the blood pressure that is tried anesthetized dog to be influenced, and does not relatively have significance statistical significance (P〉0.05) with matched group.Three dosage groups to the heart rate that tried anesthetized dog, electrocardiogram does not have obviously influences, and does not relatively have significance statistical significance (P〉0.05) with matched group.
Three dosage groups of compositions do not have obviously the respiratory frequency of being tried anesthetized dog and amplitude to be influenced, and does not relatively have significance statistical significance (P〉0.05) with matched group.
12, compositions is as the enforcement of antiviral drugs application
12.1, compositions has better wide spectrum preventing respiratory viruses usefulness
12.1.1, the experimentation of resisiting influenza virus in the mice Orally administered composition body
12.1.1.1, the pure Chinese medicine extract of combination of compositions system, for studying the effect of its resisiting influenza virus, adopt influenza infection mice viral pneumonia model, resisiting influenza virus effect in the Orally administered composition body is studied.
The inventor discloses total organic phenolic acid and glucosides class or the tool benzoyl that contains element sulphur from Stellaria plant Herba stellariae mediae or other a kind of Stellaria plant in the molecular structure that extracts in CN 1377680A and two parts of patents of CN 1498630A, the phenols of cinnamyl structure is the application of high polarity compound compositions such as total flavones and glucosides class thereof as antiviral drugs, comprise that test cell line proof compositions can be to adhering to the influenza virus of RNA class separately, parainfluenza virus, rhinovirus, Echovirus, herpesvirus has stronger inhibitory action with the two big class Respiroviruses such as adenovirus that belong to the DNA class, particularly to influenza virus, parainfluenza virus and adenovirus 3 have powerful inhibition, therefore have unique broad-spectrum antiviral performance.Its nasal drop is resisiting influenza virus FM in the mice animal body 1Result of the test show 50% mice protective rate (ED 50) be 0.51mg/kg, the interior therapeutic index has powerful resisiting influenza virus effect up to proof compositions more than 4500.
The composition oral administration is resisiting influenza virus FM in the mice animal body 1Result of the test because medicine is subjected to the influence etc. of digestive tract and liver first-pass effect behind the oral administration, so the therapeutic index that its therapeutic index is lower than the nasal drop administration is normal, and uses more convenient when keeping clinical effectiveness.
12.1.1.2, material and medicine
12.1.1.2.1, the animal Kunming mouse, body weight 18～22g, male and female half and half, female unpregnancy.
12.1.1.2.2, medicine is subjected to reagent: compositions dry powder (the same).
Each administration group dosage of experimental animal is 128mg/kg, 64mg/kg, 32mg/kg and 16mg/kg (pressing body surface area calculates), is equivalent to 6 times, 3 times, 2 times and 1 times of the clinical consumption per day of people respectively.Positive control drug: SHUANGHUANGLIAN KOUFUYE, Harbin Pharmaceutical Group's three smart pharmaceutcal corporation, Ltd products, specification: 10ml/ props up, lot number: 03051841, the experimental animal dosage is 9ml/kg (pressing body surface area calculates), is equivalent to 9 times of clinical day use amount of people.Ribavirin, Sichuan Mei Dakang pharmaceutcal corporation, Ltd product, specification: 100mg/ sheet, lot number: 030212.The experimental animal dosage is 106mg/kg (pressing body surface area calculates), is equivalent to 9 times of clinical day use amount of people.Above medicine all is mixed with desired concn with distilled water before use.
12.1.1.2.3, the suitable strain (FM of viral influenza virus A 1 type Mus lung 1), draw from national influenza center, by this institute virus research chamber preservation strain that goes down to posterity.Hemagglutinative titer E=1:1280.
12.1.1.3, method and result
12.1.1.3.1, FM 1Increase poison and preserve
Get 9 age in days chick embryo allantoic cavities inoculation FM 1Seed culture of viruses liquid, 34 ℃ of conventional cultivations, 48 hours collection allantoic fluids.Collect hemagglutinative titer with the Microhemagglutination method〉allantoic fluid of 1:1280, the tubule packing, it is standby to put-70 ℃ of refrigerators.
12.1.1.3.2 the mensuration (LD of influenza virus Mus lung adapted strain virulence 50)
Get 30 of regular grade Kunming mouses, body weight 18～22g, male and female half and half, female unpregnancy.View volume is heavy to be divided into 5 groups with sex with the mice stratified random, and 6 every group, wherein male and female are each 3.Under the absolute ether light anaesthesia, use FM 150ul/ collunarium infecting mouse of the suitable influenzae strain virus liquid of Mus lung, each dilution factor of organizing mouse infection virus is respectively 1,10 -1, 10 -2, 10 -3, 10 -4After the mouse infection virus, observe and write down each treated animal death toll day by day, observed continuously 14 days.According to the dead sum of each treated animal, press the median lethal dose(LD 50) (LD that the Reed-Muench method is calculated mouse infection virus 50).
The results are shown in Table 4. animal dead percentage rate computer charts
Table 4. animal dead percentage rate computer chart
Figure A200710130829D00181
*Denominator is infecting mouse sum (comprising dead, alive Mus) ▲Molecule is dead number of mice
As seen from Table 8: the mortality rate of each dilution factor infecting mouse is respectively 100%, 62.5%, 22.22%, 0% and 0%., can make the dilution factor of half infecting mouse death should be 10 -0～10 -2Between and calculate LD 50Value.
The first step: by formula distance proportion=50% per cent death loss-50/ 50% per cent death loss-＜50% per cent death loss, in will showing with LD 50(55%) and following (22%) adjacent several substitution formula are tried to achieve distance proportion number=62.5-50/62.5-22.22=0.31 on the value
Second step: will the logarithm of the viral dilution degree of 50% animal dead, add the distance proportion number, multiply by the logarithm of the coefficient of dilution again, promptly try to achieve LD 50 value.The logarithm of viral dilution degree of 50% animal dead is 10 -1
LD 50=1+0.31 * 1 (logarithm of the coefficient of dilution 10)=1.31
According to result of calculation, try to achieve the mice collunarium and suck infection FM 1LD 50Be 10 -1.31
12.1.1.3.3, compositions is to the therapeutical effect of influenza infection induced mice viral pneumonia
Get 80 of regular grade Kunming mouses, body weight 18～22g, male and female half and half.Press dosage shown in the table 1, the apparent motion rerum natura not and body weight the mice stratified random is divided into 8 groups, 10 every group, male and female half and half.Under the absolute ether light anaesthesia, with 5 times of LD 50Suck and infect 7 groups of mices with batch Embryo Gallus domesticus seed culture of viruses influenza virus liquid 50ul collunarium that goes down to posterity, the normal control group splashes into the equivalent normal saline solution.Each administration treated animal gives gastric infusion in the time of viral infection, every day 2 times, and each 0.4ml/20g body weight, successive administration 6 days, normal control group and virus control group are all irritated stomach and are given the isometric(al) distilled water.From the self-infection virus, observe and write down each experimental group dead animal number day by day, observed continuously 14 days.Sum with every treated animal death in 14 days calculates mortality rate (survival rate), dead protective rate and mean survival time by following formula.Mortality rate (survival rate)=survival (death) number/laboratory animal number * 100%; Dead protective rate (%)=(matched group mortality rate-experimental group mortality rate) * 100%; Every animal survival number of days addition/laboratory animal number in The average survival time day=on the same group; Prolong vital rates (%)=(experimental group on average survive natural law-matched group on average survive natural law)/matched group natural law * 100% of on average surviving.X 2, t check comparable group differences.The results are shown in Table 5, table 6:
Table 5. composition solution is to the influence of the dead protective rate of influenza virus infecting mouse
Figure A200710130829D00192
Figure A200710130829D00201
Table 6. composition solution is to the influence of influenza virus infecting mouse mean survival time
Figure A200710130829D00202
Figure A200710130829D00211
Compare with normal group: ▲ ▲P＜0.01; Compare with the virus group: *P＜0.05, *P＜0.01
Table 5 shows with table 6 result: virus control group mice is substantially all dead about 5 days behind the viral infection, and after giving the composition solution of various dose, all can reduce the mortality rate of infecting mouse, make and obviously prolong infection animal The average survival time day (very remarkable) with 128mg/kg, 64mg/kg dosage group.Its effect can strengthen along with the increasing of dosage within the specific limits.Compare with the virus control group, remarkable statistical significance is arranged.Composition solution 64mg/kg, 128mg/kg dosage group reduce the mortality rate of infecting mouse and prolong its mean survival time effect close with ribavirin tablet, obviously are better than the SHUANGHUANGLIAN KOUFUYE group.
12.1.1.3.4, compositions causes mice viral pneumonia model pneumonopathy to influenza infection and becomes influence:
Get 80 of healthy regular grade Kunming mouses, body weight 18～22g, male and female half and half.Press dosage shown in the table 11, the apparent motion rerum natura not and body weight the mice branch deposited be divided into 8 groups at random, 10 every group, male and female half and half.Under the absolute ether light anaesthesia, with 5 times of LD 50Suck and infect 7 groups of mices with batch Embryo Gallus domesticus seed culture of viruses influenza virus liquid 50ul collunarium that goes down to posterity, normal control group collunarium sucks the equivalent normal saline.Each administration treated animal gives gastric infusion respectively in viral infection, every day 2 times, and the each 0.4ml/20g body weight of administration volume, successive administration 4 days, normal control group and virus control group are all irritated stomach and are given the isometric(al) normal saline.Infection animal is isolated nursing, freely ingests drinking-water.After the last administration 1 hour, the animal back of weighing is fixing, cut off skin of chest, cut off left oxter arterial blood letting and put to death animal.The mice thoracic cavity is opened in dissection, takes out full lung, rejects trachea and lung
After the lymphoid tissues such as door, blot the lungs surface liquid with filter paper, put balance and weigh, heavily calculating the lung exponential sum one by one divided by body weight respectively organizes lung index suppression ratio by lung.With t check comparable group differences.Leave and take lung tissue sample, carry out tectology and observe and take the photograph sheet.The results are shown in Table 7..
Table 7. composition solution is to the pathogenic toxicity pneumonia lung index influence of influenza virus infecting mouse
▲ ▲Compare with normal group; *Compare with the virus group
By table 7 result as seen: compare with normal group, the lung index of virus control group mice obviously increases, and statistical significance is extremely remarkable.After giving the composition solution of various dose, infection animal lung weight in wet base index all is starkly lower than virus control group (P＜0.01～0.05), and its therapeutic effect can be along with the increase of drug dose, and the lung exponential quantity also reduces accordingly, presents better correlation.Composition solution 128mg/kg, 64mg, 32mg/kg dosage treated animal therapeutic effect are better than ribavirin tablet, significantly are better than the SHUANGHUANGLIAN KOUFUYE group.
12.1.2, oral administration is used in influenza patient's clinical practice effect:
Get the compositions of 1.32g and load into 6 of capsules with No. 1 capsule, each oral one, can in 1 day half, cure a routine influenza patient every day 3 times, the fastest only needs to cure half a day.The viral influenza of clinical personal treatment has 100 many cases time over 2 years, no matter is which day in the influenza course of disease, in case used compositions about 1～2 two day, control clinical symptoms and then reached the purpose of curing influenza, but and operate as normal with learnt; Comprise that the back of taking medicine can eliminate headache in more than ten minute to 30 minutes fully, then stuffy nose relieving, eliminate clear nasal mucus, tear and symptom such as pain from head to foot, brought down a fever in 36 hours at 24 hours.The early stage result of use of falling ill is good more, the fastest case once morbidity that night (headache, from head to foot pain, 39.5 ℃, drop tears, nasal mucus etc.) use medicine just before going to bed one time, only 1, be that operate as normal has been learnt the next morning.Compositions prevents and treats viral influenza to reach efficient even specific drug degree.
12.2, the animal vivo test and the result thereof of oral and intravenous administration show that also compositions can obviously reduce hepatitis B virus DNA in the animal body, also has obvious inhibition effect to hepatitis virus:
12.2.1, medicine: the pale brown color medicated powder of compositions is prepared with normal saline.
Positive control medicine lamivudine is by the plain Kang Wei of Ge Lan drugmaker product, and lot number: B008923 prepares with normal saline.
12.2.2, hepatitis virus: dhbv dna DNA (DHBV-DNA) strong positive serum ,-70 ℃ of preservations.
12.2.3, animal: 1 age in days Beijing duck, available from progressive species duck animal feeding field, Beijing.
12.2.4, reagent: α -32P-dCTP is available from the auspicious biotechnology of Beijing good fortune engineering company, and the nick translation medicine box is available from Pu Luomaige company (Promega Co.); Sephadex G-50, Ficoll PVP is available from Sweden Pharmacia company; SDS West Germany Merck company product; Milt DNA, bovine serum albumin are that Instite of Biophysics, Chinese Academy of Sciences produces; Nitrocellulose filter 0.45 μ m Amersham produces.
12.2.5.1, experimental technique: the duck hepatitis B virus infection method
1 age in days Beijing duck is clear through the positive Sanguis Anas domestica of lower limb shin intravenous injection Shanghai sheldrake DHBV-DNA, and every 0.2ml got blood in back 7 days in infection, separation of serum, and-70 ℃ of preservations are to be checked.
12.2.5.2, Drug therapy test:
DHBV infect duckling after 7 days random packet carry out the Drug therapy test, 6 every group, 2 dosage groups of administration component, oral administration is respectively 250mg/kg and 500mg/kg, the intraperitoneal injection group is respectively 25mg/kg and 50mg/kg group, 1 day 2 times, administration 10 days.
If virus control group (DHBV) is with the physiologic saline for substitute medicine.The positive drug lamivudine, oral administration 50mg/kg, 1 day 2 times, administration 10 days.
The 7th day is (TO) before the medication after infection, and medication the 5th day (T5) after medication the 10th day (T10) and the drug withdrawal the 3rd day (P3), is got blood from duck lower limb shin vein, and separation of serum is to be checked in-70 ℃ of preservations.
12.2.5.3, detection method:
It is clear to get above-mentioned Sanguis Anas domestica to be checked, every batch with the time point film, measure the dynamic of the clear middle DHBV-DNA level of Sanguis Anas domestica, press nick translation medicine box description method,, and make the clear dot blot hybridization of Sanguis Anas domestica with 32P labelling DHBV-DNA probe, autoradiography diaphragm speckle, measure OD value (optical filter is 490nm) at enzyme mark detector, calculating serum DHBV-DNA density, with hybridization spot OD value as specimen DHBV-DNA level value.
12.2.5.4, drug effect calculates:
12.2.5.4.1, calculate the meansigma methods (X ± SD) of every group of duck different time serum DNA OD value, and with (T0) OD value comparison before the 3rd day (P3) serum DHBV-DNA level after different time (T5, T10) and the drug withdrawal after every group of duck medication and the administration on the same group, adopt paired t-test, meter t1, P1 value.Analyze the significance of difference, judge the inhibition effect of medicine viral infection.
12.2.5.4.2, calculate after every group of duck medication the 3rd day (P3) serum DHBV-DNA suppression ratio behind the different time (T5, T10) and drug withdrawal, the dynamic of the clear DHBV-DNA suppression ratio of Sanguis Anas domestica respectively organized in and mapping.
Figure A200710130829D00231
12.2.2.4.3, with identical with the virus control group respectively time D HBV-DNA suppression ratio of drug treatment group different time DHBV-DNA suppression ratio relatively, adopt t check in groups, take statistics to learn and handle, calculate t2, P2 value, analyze the significance of difference, the judgement drug effect.
12.2.2.5, the result:
12.2.2.5.1, after 1 age in days Beijing crow infects DHBV, serum DHBV-DNA is dynamic
The DHBV-DNA results of dot sees Table 1 behind the DHBV-DNA infected duck oral normal saline.36 serum DHBV-DNA of experimental infection duck total positives.6 ducklings of virus control group infect the 7th day serum DHBV-DNA total positives in back, test in omnidistance 21 days steady substantially behind the serum DHBV-DNA level infection.
12.2.2.5.2, the positive drug lamivudine is to the influence of DHBV-DNA
The oral positive control drug lamivudine of DHBV infected duck 50mg/kg, 1 day 2 times, administration 10 days.The results are shown in Table 1, table 2.Pair analysis, the OD value comparison of (TO) before administration the 10th day (T10) and the administration, descending has highly significant (P＜0.01), to serum DHBV-DNA suppression ratio and virus control group, becomes group analysis administration the 10th day (T10) that significant differences (P＜0.01) is arranged after the administration.3 days (P3) not statistically significants have rebound phenomenon after the drug withdrawal.
12.2.2.5.3, compositions in DHBV infected duck body to the experimental result of the influence of the clear DHBV-DNA of Sanguis Anas domestica:
Two kinds of experiment branch oral administration and intraperitoneal injections are selected two dosage groups respectively for use, and oral administration is respectively 100mg/kg, 200mg/kg, 1 day twice, administration 10 days (Bid * 10); Intraperitoneal injection is 15mg/kg, 30mg/kg, and 1 day 2 times, administration 10 days (Bid * 10) is observed medicine to toxicity of duck and the influence of the clear hepatitis B virus DNA of Sanguis Anas domestica (DHBV-DNA) d.
12.2.2.5.3.1, oral administration experiment shows the heavy dose of group of compositions oral administration 200mg/kg, 1 day twice, avirulence, by pairing statistics, after the administration of 200mg/kg group the 5th day, the 10th day, the clear DHBV-DNA of treatment group Sanguis Anas domestica had highly significant descend (P＜0.01).By statistics in groups and matched group separately relatively, after the administration the 5th day, after 10 days and the drug withdrawal 3 days, the clear DHBV-DNA of treatment group Sanguis Anas domestica all had highly significant to descend (P＜0.01,0.01), and its therapeutic index is 486, and this product is not seen the rebound phenomenon after the drug withdrawal.
12.2.2.5.3.2, intraperitoneal injection, heavy dose of group 30mg/kg group, twice administration in 1 day 10 days, by after the administration of pairing statistics the 5th day, the clear DHBV-DNA of treatment group Sanguis Anas domestica promptly had highly significant descend (P＜0.01).Statistics and matched group separately relatively have highly significant descend (P＜0.01,0.01) in groups.15mg/kg group, by the pairing statistics, after the drug withdrawal 3 days, the clear DHBV-DNA of treatment group Sanguis Anas domestica had remarkable decline (P＜0.01).By statistics in groups and matched group separately relatively, after the administration after the 5th day and the drug withdrawal 3 days, the clear DHBV-DNA of treatment group Sanguis Anas domestica had remarkable decline (P＜0.01), and its therapeutic index is 1288, did not see the rebound phenomenon after the drug withdrawal.
It is better, the prior advantage of this product than positive control drug lamivudine etc. that oral administration and intraperitoneal injection there is no rebound phenomenon.
12.3, the beneficial effect of compositions also can provide by following 2 reports and clinical practice result:
12.3.1, report 1. test cell lines proof compositions can effectively suppress HIV (human immunodeficiency virus)-1 (HIV-1)
The result:
In vitro tests shows that compositions has the effect of anti AIDS virus (HIV-1), and the result is as follows:
Drug level (mg/ml) 0.20 0.10 0.05 0.025
Suppression ratio (%) 98.58% 64.3400% 23.90% 12.11%
Calculate IC by statistics 50=0.113mg/ml
Positive control drug AZT concentration is when 1g/ml, and suppression ratio is 100%.
12.3.1.2, cell toxicity test result is as follows:
Drug level (mg/ml) 5 2.5 1.25 0.625
Cell mortality (%) 72 40.04 14.88 0
12.3.1.3, calculate TC by statistics 50So=3.235mg/ml is safety index (TC 50/ IC 50) be 29.09.
12.3.2, report 2. test cell lines proof compositions can effectively suppress the result of H 5 N 1 avian influenza type virus:
12.3.2.1, medicine: " compositions " is Herba stellariae mediae extract.
12.3.2.2, virus: the H 5 N 1 avian influenza type
12.3.2.3, cell: the human embryonic lung cell
12.3.2.4, the result: what the nontoxic boundary of compositions pair cell was selected the results are shown in Table 8.:
The nontoxic boundary of table 8. compositions pair cell is selected (drug level mg/ml)
Figure A200710130829D00251
The result: compositions is non-toxic concn with 1mg/ml
12.3.2.5, compositions really sees Table 9. to the toxic effect that presses down of H 5 N 1 avian influenza type virus in the test cell line:
Table 9. compositions presses down toxic effect really to bird flu virus
Figure A200710130829D00252
Its method is: virus is with 100TCID 50Be infective dose, first infection cell monolayer, dosing after 3 hours.
12.3.2.6, conclusion: " compositions " has the effect of strong inhibition H 5 N 1 avian influenza type virus on cell in vitro is cultivated with 1mg/ml concentration.
12.3.3, the actual efficacy of personal composition oral administration clinical treatment influenza:
12.3.3.1, method:
Compositions is made every tablet of " buccal tablet " that contains compositions 50mg, and per hour containing is 1, administration every day 10 times, two days common administration 1g.
12.3.3.2, the result: " buccal tablet " of set of applications compound treatment 11 routine influenza patients the results are shown in Table 10.:
The clinical personal example of table 10. to 50mg/ treatment 11 routine influenza patients of compositions " buccal tablet "
Figure A200710130829D00261
12.3.3.3, conclusion: compositions is clinical gives " buccal tablet " treatment 11 routine influenza patients, all can in two days, cure, can bring down a fever and eliminate flu-like symptom in the fastest half a day that only needs, thereby can go to school or go to work, compositions can shorten viral influenza course of treatment more than 5 days at least, therefore, has the function of extraordinary prevention and treatment influenza, it is evident in efficacy to be better than influenza virus neuraminidase inhibitors such as English, beautiful Zanamivir and Oseltamivir, can reach the specific drug degree.
12.3.4, the actual efficacy of the clinical personal treatment herpes simplex of compositions and herpes zoster:
12.3.4.1, method: suffer from herpes simplex and taped blister examine disease human compositions varnish and constantly be applied in the affected part;
12.3.4.2, the result: see Table 11.:
The mucilage of the clinical personal 0.5g/25ml of table 11. or varnish are treated the example of 10 routine herpes simplexes and herpes zoster
Figure A200710130829D00262
12.3.4.2, conclusion: from the patient of visible clinical use combination treatment herpes simplex of table 16. and herpes zoster, thus its dosage between 0.2g～0.5g, and can pain relieving in 1 to 3 day, the bleb that disappears, incrustation reach recovery from illness, reaches the curative effect of feeling quite pleased.Wherein drug concentrations and herpes area size has substantial connection with curative effect.
12.3.5, the result of implementation of compositions clinical treatment infantile parotitis:
12.3.5.1, method: get compositions and make every tablet of " buccal tablet " that contains compositions 50mg, per hour containing is 1, administration every day 3 times, two days administration 0.3g altogether.
12.3.5.2, the result of implementation of clinical personal treatment infantile parotitis sees Table 12.:
The clinical personal example for the treatment of 10 routine parotitis children of table 12. to buccal tablet 50mg/ sheet
12.3.5.3, conclusion: the result of implementation of compositions clinical treatment infantile parotitis as can be seen behind buccal tablet 50mg * 3 slice/day of the clinical use compositions of parotitis sick child, only needs can fully recover in about 2 days, can obtain the curative effect of specific drug.
12.3.6, the actual efficacy of the viral sexually transmitted disease condyloma of compositions clinical treatment:
12.3.6.1, method: during the viral sexually transmitted disease condyloma of compositions clinical treatment, adopt method oral and that external combines, the promptly clinical 0.25g/cm that gives 2Membrane and give oral treatment of tablet that content is 0.1g, obtain better effect.
12.3.6.2, curative effect: see Table 13.:
The clinical personal example that gives membrane and tablet in treatment 7 routine condyloma acuminatum patients of table 13.
Figure A200710130829D00281
12.3.6.3, conclusion: the compositions clinical practice is treated viral sexually transmitted disease (STD) " condyloma acuminatum " and is obtained the effect of making us more satisfied equally.
12.3.7, compositions is applied to the example of flu-prevention, parotitis etc. as " disinfectant ".
12.3.7.1, method: " disinfectant " of using compositions as: aerosol spray is sprayed in the oral cavity, collunarium collunarium, oral chewing gum or liquid chewing gum etc. in nasal cavity.
12.3.7.2, the result: compositions " disinfectant " flu-prevention and parotitis effect see Table 14.:
Table 14. compositions " disinfectant " is used for the example of flu-prevention, parotitis etc. certainly
Figure A200710130829D00282
12.3.7.3, conclusion: " disinfectant " that the aerosol made from compositions, nasal drop, chewing gum, liquid chewing gum etc. are applied as the prevention Respirovirus obtains the effect of making us more satisfied.

Claims (14)

1. broad-spectrum antiviral compositions, it is characterized in that from the plant Herba stellariae mediae or from a kind of molecular weight distribution of other a kind of Stellaria plant extract 2,000 dalton～900,000 dalton, the phenolic acid sexual element of total sulfuric ester peptidoglycan class that is dark brown black and Herba stellariae mediae flavone are by 55%～90% to 45%～10% compositions of forming;
Total peptide partly accounts for 15% to 25% and by comprising that following 17 seed amino acids are respectively by its composition of proportions: aspartic acid: 12.118%～13.18% in the chemical constitution of the phenolic acid sexual element of this total sulfuric ester peptidoglycan class, glutamic acid: 13.036%～18.425%, serine: 4.156%～6.19%, histidine: 1.89%～1.946%, glycine: 8.909%～9.915%, threonine: 4.837%～7.129%, arginine: 2.92%～3.677%, alanine: 6.05%～9.647%, tyrosine 2.596%～2.877%, cystine: 1.89%～0.2%, valine: 5.94%～7.577%, methionine: 0.2%～2.09%, phenylalanine: 2.74%～3.738%, isoleucine: 4.28%～5.046%, leucine: 4.70%～5.53%, lysine: 5.77%～7.965%, proline: 4.61%～8.60%;
The content of element sulphur is 0.5～5% in the chemical constitution of the phenolic acid sexual element of this total sulfuric ester peptidoglycan class, except that institute's sulfur-bearings such as cystine, methionine, remaining sulfur and polysaccharide form polysaccharide sulfate or/and with sulfydryl or/and exist with the form of thioether;
Sugar moieties in the chemical constitution of the phenolic acid sexual element of this total sulfuric ester peptidoglycan class mainly is the total polysaccharides that is made of glucose, galactose and arabinose, present the pale brown colour response of sugar and two maximum absorption bands occur at 400nm～410nm and 480nm～490nm with phenol-strong sulfuric acid response, wherein the peak of 400nm～410nm is that main peak doubles than the absworption peak that 480nm～490nm occurs; Record total polysaccharides through phenol-concentrated sulphuric acid method and in its chemical constitution, account for 55%～65%;
The phenolic acid sexual element of this total sulfuric ester peptidoglycan class and ferric chloride test solution manifest a kind of dark green dark brown reaction of phenols, and at 220nm～235nm place and 330nm～two maximum absorption bands of 350nm place appearance, wherein 220nm～235nm peak is a main peak, peak height than 330nm～350nm goes out one times, and the content of phenol is 10%～15% in its structure;
2. the described a kind of broad-spectrum antiviral compositions of claim 1 is characterized in that and can extract from the plant Herba stellariae mediae or from following 78 kinds of Stellaria plants: Stellaria alsine Grim., brown lobe Stellaria alsine Grim., the Anhui Herba stellariae mediae, blunt calyx Herba stellariae mediae, the apicule Herba stellariae mediae, Herba malachii aquatici, the husky Herba stellariae mediae of giving birth to, the Alishan Herba stellariae mediae, slender lobule Alishan Herba stellariae mediae, the north Herba stellariae mediae, short lobe Herba stellariae mediae, long lobe Herba stellariae mediae, the northeast Herba stellariae mediae, China's Herba stellariae mediae, the Taiwan Herba stellariae mediae, thick leaf Herba stellariae mediae, wrinkle leaf Herba stellariae mediae, David's Herba stellariae mediae, the Herba stellariae mediae for sleeping in of laying down, the needle-like Herba stellariae mediae for sleeping in of laying down, the southwest Herba stellariae mediae, Stellaria dianthifolia Williams, the forked cyme Herba stellariae mediae, the bifid Herba stellariae mediae, narrow leaf bifid Herba stellariae mediae, line leaf bifid Herba stellariae mediae, Herba stellariae mediae looses in the shop, turn over white Herba stellariae mediae, Stellaria diversiflora Maxim., the different colored Herba stellariae mediae of naked stamen, the concave veins Herba stellariae mediae, the Du Shi Herba stellariae mediae, line stem Herba stellariae mediae, line handle Herba stellariae mediae, spend more Herba stellariae mediae, standing grain leaf Herba stellariae mediae, dredge pubescence standing grain leaf Herba stellariae mediae, turn green standing grain leaf Herba stellariae mediae, Jiangzi's Herba stellariae mediae, the XIACAO Herba stellariae mediae, Stellaria henryi williams, Xingan's Herba stellariae mediae, the introversion Herba stellariae mediae, the slender lobule Herba stellariae mediae, Stellaria maximowixziana Franch., Stellaria micrantha Hayata, gentle Herba stellariae mediae, goose intestinal Herba stellariae mediae, Herba stellariae mediae is given birth in new natural pond, eight stamen Herba stellariae mediaes, the raspberry Herba stellariae mediae, Herba stellariae mediae is given birth in the natural pond, exhibition leaf Herba stellariae mediae, handle flower Herba stellariae mediae, handle leaf Herba stellariae mediae, Herba stellariae mediae becomes mildewed, imitation stone is given birth to Herba stellariae mediae, blinks, flint lobe Herba stellariae mediae, net arteries and veins Herba stellariae mediae, the rock Herba stellariae mediae, stone is given birth to Herba stellariae mediae, embrace stem stone and give birth to Herba stellariae mediae, accurate Ge Er Herba stellariae mediae, the Su Shi Herba stellariae mediae, star hair Herba stellariae mediae, garden calyx Herba stellariae mediae, intend umbrella flower Herba stellariae mediae, little fine hair Herba stellariae mediae, three type Herba stellariae mediaes, the Turkestan Herba stellariae mediae, the wetland Herba stellariae mediae, umbrella flower Herba stellariae mediae, the LVHUA Herba stellariae mediae, Stellaria wushanensis Williams, five Herba stellariae mediaes, Yunnan Herba stellariae mediae or cattle Stellaria plant cattle Herba stellariae mediae.
3. the described preparation of compositions method 1 of claim 1, its step comprises: crude drug or the bright grass of plant Herba stellariae mediae or other a kind of Stellaria plant are converted to Radix Glycyrrhizae, pulverize the back and obtain aqueous solution through precipitation with water extraction, centrifugal or filter the back by the macroporous resin column handled well in advance to adsorb the composition of described compositions, wash macroporous resin column, again macroporous resin column is heated and after 45 ℃～55 ℃, wash post and collect effusive dark brown distilled water liquid with the thermal distillation of same temperature, continue with heating to 45 ℃～55 ℃, concentration is at 20%～35% lower alcohols or acetone eluting from the resin column, collect brownish red or dark reddish brown tryptophol liquid or acetone solution that eluting obtains, to merge with the dark brown water liquid of collecting behind this solution recovery alcohol or the acetone, be concentrated into than weighing and left standstill 12 hours after 0.80 to 1.00, centrifugal removal impurity, adding concentration and be 95% ethanol makes in the solution alcohol or acetone concentration reach 50%～70%, leave standstill, leaching precipitate and once with 95% washing with alcohol of 3 times of volumes, the washing with acetone of 2 times of volumes once, through 60 ℃ of vacuum dryings or be dissolved in water into proportion be 1.00～1.20 solution again spray drying promptly get compositions; Wherein lower alcohols comprises methanol, ethanol, propanol and isopropyl alcohol.
4. the described preparation of compositions method 2 of claim 1, its step comprises: the back is cleaned, pulverized to bright grass or the bright Radix Glycyrrhizae of plant Herba stellariae mediae or other a kind of Stellaria plant extract aqueous solution, with this aqueous solution through precipitation, centrifugal or filter after obtain liquid; This liquid is adsorbed on described compositions composition on the post through the anionite post, with the 2N sodium chloride solution described compositions composition is eluted from post, again through desalting processing, be concentrated into than weighing and left standstill 12 hours after 0.80 to 1.00, centrifugal removal impurity, add concentration and be 95% ethanol and make that concentration of alcohol reaches 50%～70% in the solution, leave standstill, leaching precipitate and with 95% washing with alcohol of 3 times of volumes 1～2 time, the washing with acetone of 2 times of volumes 1～2 time is through 60 ℃ of vacuum dryings or to be dissolved in water into proportion be that 60 ℃ of vacuum dryings or spray drying promptly get compositions again for 1.10～1.20 solution;
Wherein the anionite post comprises weak anion exchange resin post and cellulose anion exchanger post.
5. the described preparation of compositions method 3 of claim 1, its step comprises: after the dried pulverizing medicinal materials with plant Herba stellariae mediae or other a kind of Stellaria plant, with twice of the flooding of 5～10 times of volumes, each 1～3 hour, the dark-brown extracting solution that united extraction obtains, be evaporated to proportion 0.7～0.9, left standstill 12 hours, the filtering precipitation, adding 95% lower alcohols or acetone make concentration reach 60%～70% in the solution, left standstill 12 hours, the precipitate that the filter collection is separated out promptly gets the compositions crude product, this compositions crude product is dissolved after high speed centrifuge 15 with an amount of pure water, 000rpm, centrifugal 15 minutes, clear liquor elder generation via hole diameter after centrifugal is that the ceramic membrane treatment facility fine straining of 0.22 μ m is handled, be ultrafilter membrane ultrafiltration more than 2000 through molecular weight cut off again, will be through the solution concentration after part small-molecular weight impurity is removed in ultrafiltration to proportion 0.7～0.9, adding lower alcohol or acetone makes concentration reach 85% to carry out the secondary precipitate with ethanol, leave standstill the sucking-off supernatant after 12 hours, precipitate is closely dried through decompress filter, in time successively add an amount of dehydrated alcohol and washing with acetone, sucking filtration is extremely done, and promptly gets purer powdered composition through 60 ℃ of vacuum dryings immediately; Wherein lower alcohol is methanol, ethanol, propanol, isopropyl alcohol;
6. the pharmaceutical formulation that can be made into of the described compositions of claim 1 is as follows: injection, injectable powder, aerosol, buccal tablet or chewable tablet, capsule, drop pill, micropill, microcapsule, nasal drop or nasal spray, membrane or varnish, mucilage, water preparation and chewing gum etc.
7. the application of the described compositions of claim 1 in preparation treatment HIV (human immunodeficiency virus) (Human Immunodeficiency Virus) medicine.
8. the described compositions of claim 1 is preparation treatment influenza virus, comprises the application in the medicine of bird flu virus.
9. the application of the described compositions of claim 1 in preparation treatment herpesvirus medicament.
10. the application of the described compositions of claim 1 in preparation treatment hepatitis virus medicament.
11. the application of the described compositions of claim 1 in preparation treatment mumps virus medicine.
12. the application of the described compositions of claim 1 in preparation treatment condyloma acuminatum medicine.
13. the application of the described compositions of claim 1 in the antiviral disinfectant of preparation.
14. the application of the described compositions of claim 1 in the antiviral health product of preparation.
CNA2007101308293A 2007-08-22 2007-08-22 High-efficient antiviral medicament composition in chickweed as well as preparation method and use thereof Pending CN101371861A (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2447895C2 (en) * 2010-05-14 2012-04-20 Государственное образовательное учреждение высшего профессионального образования "Сибирский государственный медицинский университет Федерального агентства по здравоохранению и социальному развитию" (ГОУ ВПО СибГМУ Росздрава) Antioxidant hepatoprotector and method of preparing
CN102512479A (en) * 2010-02-03 2012-06-27 朱耕新 Chickweed water extract, its preparation method, and liquid chromatography method for determining content, fingerprint spectrum and molecular weight distribution as well as application thereof
CN111150724A (en) * 2020-01-16 2020-05-15 华中农业大学 Application of valine in preparation of medicine for treating or preventing avian influenza virus infection

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102512479A (en) * 2010-02-03 2012-06-27 朱耕新 Chickweed water extract, its preparation method, and liquid chromatography method for determining content, fingerprint spectrum and molecular weight distribution as well as application thereof
CN102512479B (en) * 2010-02-03 2015-04-15 朱耕新 Chickweed water extract, its preparation method, and liquid chromatography method for determining content, fingerprint spectrum and molecular weight distribution as well as application thereof
RU2447895C2 (en) * 2010-05-14 2012-04-20 Государственное образовательное учреждение высшего профессионального образования "Сибирский государственный медицинский университет Федерального агентства по здравоохранению и социальному развитию" (ГОУ ВПО СибГМУ Росздрава) Antioxidant hepatoprotector and method of preparing
CN111150724A (en) * 2020-01-16 2020-05-15 华中农业大学 Application of valine in preparation of medicine for treating or preventing avian influenza virus infection

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