CN101670074A - Preparation method for antivirus compound - Google Patents
Preparation method for antivirus compound Download PDFInfo
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Abstract
The invention provides a preparation method for an antivirus compound which adopts a supercritical carbon dioxide extraction method to extract a lipophilic active ingredient of patchouli and grass-leaved sweetflag, which is characterized in that the lipophilic active ingredient is volatile oil, the extracting efficiency is more than two times of the efficiency of steam distillation, no solvent toxicity remains, heat sensitive ingredient in natural active ingredient is not easy to de-compound and damage, and the natural characteristic of the extraction can be furthest maintained. Furthermore, active parts in antivirus compound are obtained by respectively exacting with water and alcohol, the extraction process is simple, scientific and reasonable, and active ingredient in medicinal materials can be furthest extracted. After impurities are removed, the active ingredient contents of mangiferin and phillyrin in medicine are respectively 16 time and 2 times higher than the same extracted with a prior method.
Description
Technical field
The present invention relates to the field of Chinese medicines, be specifically related to a kind of antivirus compound formulation.
Background technology
" Ministry of Health of the People's Republic of China's new drug become a full member standard " WS3-49 (X-39)-92 (z) discloses a kind of antivirus oral liquid, this oral liquid is prepared through water extract-alcohol precipitation by Gypsum Fibrosum, Radix Isatidis, Rhizoma Phragmitis, Radix Rehmanniae, Radix Curcumae, the Rhizoma Anemarrhenae, Rhizoma Acori Graminei, Herba Pogostemonis and Fructus Forsythiae nine flavor Chinese medicines, be widely used in the viral infection illness such as upper respiratory tract infection, influenza and mumps that the heating of anemopyretic cold, epidemic febrile disease causes, have clinical effectiveness preferably.But because traditional simple and crude this product active constituent content that causes of preparation technology is low, particularly volatile oil component, and this product is a liquid preparation, is difficult for storing, and carries also inconvenient.
State Intellectual Property Office disclosed the application for a patent for invention of " preparation method of concentrated anti-virus oral administration liquid " (publication number is CN1899571A) on 01 24th, 2007, this application is improved the traditional preparation process of above-mentioned oral liquid, adopt traditional method to concoct the Chinese crude drug raw material, adopt high speed centrifugation and membrane separation technique that drug level is doubled in unit volume then, content of effective is the twice of original same dose.The application for a patent for invention of one " new technology of preparation antivirus oral liquid " (publication number is CN1939525) was disclosed on 04 4th, 2007, this application discloses the method that the volatile oil in the above-mentioned oral liquid is prepared into clathrate, has improved volatile oil content of effective in the antivirus oral liquid.Though above-mentioned two disclosed preparation methoies of patent application are improved on the basis of traditional method, but still continue to use original aqueous extraction-alcohol precipitation technology, loss of effective components is bigger in the leaching process, products obtained therefrom still can't overcome the low shortcoming of traditional product active constituent content, not only antiviral drug effect is not in full use, cause the bigger wasting of resources, make that also patient's dosage is big.Secondly, the scheme of above-mentioned two patent applications is resulting to remain traditional liquid dosage form, be inconvenient to carry, and shelf life of products is short.
Summary of the invention
The problem to be solved in the present invention is to improve content of effective in the anti-virus compound.
The technical scheme that the present invention solves the problems of the technologies described above is:
A kind of preparation method of antiviral composition, this method comprises following step:
(1) get Herba Pogostemonis 2.9 weight portions, Rhizoma Acori Graminei coarse powder 2.5 weight portions, CO packs into
2In the supercritical extraction reactor, extracted 2 hours down for 55 ℃, collect volatile oil, stay medicinal residues standby in pressure 28MPa, temperature;
(2) get the Rhizoma Anemarrhenae 2.5 weight portions, Gypsum Fibrosum 5.7 weight portions, Rhizoma Phragmitis coarse powder 6.1 weight portions, add 8～10 times in water, 80 ℃ are extracted 2～3 times down, each 1～2 hour, filter, merge extractive liquid,, temperature be 100 ℃ of vacuums be-0.08～-0.04MPa under concentrating under reduced pressure become clear paste, making its density under 50 ℃ is 1.05～1.15g/mL;
(3) getting Fructus Forsythiae 4.6 weight portions, Radix Isatidis 12.9 weight portions, Radix Curcumae 2.5 weight portions, Radix Rehmanniae 3.2 weight portions and step (1) gained medicinal residues mixes, extract 2～3 times with 8～10 times of 80% alcohol heating reflux, each 1～2 hour, filter, merge extractive liquid,, with ethanol extract temperature be 60 ℃ of vacuums be-0.08～-0.04MPa under concentrating under reduced pressure become clear paste, making its density under 50 ℃ is 1.05～1.15g/mL;
(4) clear paste of getting volatile oil, step (2) and (3) gained of step (1) gained is prepared into preparation commonly used according to a conventional method, as oral liquid, granule etc.
In the said method, scatter and disappear in order to prevent the described volatile oils of step (4), can be with the volatile oil beta-cyclodextrin inclusion compound before the preparation preparation, therefore, the inventive method is further comprising the steps of: the dehydrated alcohol of doubly measuring with equal-volume dilutes, splash into then in the beta-schardinger dextrin-saturated solution of 6～10 times of amount volatile oil weight,, left standstill 24 hours in 40～60 ℃ of stirrings 3～5 hours.
Compositions of the present invention preferably is prepared into granule so that preserve and carry.
In the method for the present invention, WATER AS FLOW MEDIUM, alcohol and put forward the usual practice of ratio employing this area of raw material, promptly water, pure consumption unit are volume unit; The inventive method, the prescription of the crude drug of wherein said antiviral composition is the prescription of the crude drug of the antivirus oral liquid of " Ministry of Health of the People's Republic of China's new drug become a full member standard " WS3-49 (X-39)-92 (z) defined.
The present invention is according to the difference of traditional anti-virus compound Chinese crude drug active component physicochemical property, adopt the carbon dioxide supercritical fluid extraction method to extract lipotropy effective active composition---the volatile oil of Herba Pogostemonis, Rhizoma Acori Graminei, extraction efficiency is more than 2 times of steam distillation, no solvent residue toxicity, heat-sensitive ingredients in the active skull cap components is difficult for decomposing to be destroyed, and can keep the extract natural feature to greatest extent; Adopt water, alcohol extraction to obtain active site in the anti-virus compound simultaneously respectively, extraction process is simple, scientific and reasonable, extract the effective ingredient in the medical material to greatest extent, remove the impurity composition, make medicine effective ingredient chimonin, phillyrin content in crude drug same units quality improve 16 times and 2 times than existing preparation method respectively.
Content of effective in the antiviral granule agent of the present invention can be measured by the following method:
1, the assay of chimonin and phillyrin
The preparation of reference substance solution: precision takes by weighing chimonin, phillyrin is an amount of, puts in the 50ml volumetric flask, adds methanol to scale, makes chimonin concentration 0.1172mg/ml, phillyrin concentration is 0.0480mg/ml solution, in contrast product solution.
The preparation of need testing solution: get this product 2g, precision is weighed, and puts in the tool plug conical flask, add methanol 50ml, ultrasonic 40min filters, residue adds water 10ml dissolving, with water saturation n-butanol extraction 4 times (20ml * 4), merge n-butanol layer, water bath method, residue 50% dissolve with methanol, standardize solution shakes up in the 10ml measuring bottle, and is standby.
Respectively accurate reference substance solution 10 μ l and the need testing solution 5 μ l of drawing inject high performance liquid chromatograph, mensuration, that is and, every bag contains that chimonin is not less than 2.5mg, phillyrin is not less than 0.5mg.
Measure chromatographic condition and adaptability: 4.6mm * 250mm, 5 μ m carbon, 18 bonding phase silicagel columns according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2005); 10-30: 70-90 acetonitrile-0.05mol/L potassium dihydrogen phosphate (adding 1 ‰ phosphoric acid) is a mobile phase; Detect wavelength: 277nm; Theoretical cam curve is calculated by chimonin should be not less than 5000, calculates by phillyrin and should be not less than 6000.
2, the assay of patchouli alcohol (internal standard method)
Chromatographic condition: chromatographic column is the elastic quartz post, and injector temperature is 230 ℃, and detector temperature is 250 ℃, split sampling, 70～230 ℃ of temperature programmings kept 10～15 minutes in 230 ℃, flow velocity 1.0ml/min, theoretical cam curve is calculated by patchouli alcohol should be not less than 5000;
The mensuration of correction factor: it is an amount of that precision takes by weighing AI3-28404, makes the solution of 4.0mg/ml with normal hexane, as inner mark solution, standby, it is an amount of that precision takes by weighing the patchouli alcohol reference substance in addition, puts in the 10ml volumetric flask, add n-hexane dissolution, make 0.2mg/ml solution, shake up, precision is measured in above-mentioned patchouli alcohol reference substance solution 0.2ml to the 2ml measuring bottle, the accurate inner mark solution 0.2ml that adds adds normal hexane to scale, shakes up, draw 1 μ l inject gas chromatograph, the calculation correction factor.
The preparation of need testing solution: get this product 2g, precision is weighed, and adds water 20ml dissolving, with twice of chloroform 40ml heating and refluxing extraction, each 1 hour, merge the chloroform layer, water bath method, the an amount of n-hexane dissolution of residue, be transferred to the 2ml measuring bottle, and add 100 μ L inner mark solutions, add normal hexane to the 2mL scale, shake up, standby.
The accurate need testing solution 1 μ l that draws, inject gas chromatograph is measured, that is and, every bag contains patchouli alcohol and is not less than 48 μ g.
The pharmacological action of anti-virus formulation of the present invention can prove by following pharmacodynamic experiment:
One, preventing respiratory viruses effect
1 test material
1.1 cell: mdck cell is provided by Chinese Academy of Medical Sciences's biotechnology research, and human embryonic lung diploid fibroblast (MRC-5) is provided by the Kunming zooscopy, and the A549 cell is frozen by this chamber.
1.2 cultivation composition: culture medium is DMEM (Gibco), and hyclone (Gibco), Ox blood serum are all available from Guangzhou herding factory.
1.3 virus: influenza A virus FM1, adenovirus type VII, syncytial virus (RSV), it is frozen to be this research department.
1.4 medicine: antiviral granule, press the preparation of embodiment 1 method; Antivirus oral liquid; Positive control medicine virazole (Zhaoqing, Guangzhou Xing Hu pharmaceuticals provides).
2 methods and result
2.1 drug cell toxicity test: in 96 well culture plates, add 5000--2.0 * 10
5The cell of/ml (MDCK, MRC-5, A549), 37 ℃ of following 5%CO
2In cultivate, treat that cell grows up to monolayer after, other adds the medicine to be measured of variable concentrations.Day by day the observation of cell pathological changes is cultivated and was then stopped experiment in 4-7 days.
2.2 medicine antivirus action: add each the institute's reagent thing under the non-toxic concn, four concentration of each medicine, every concentration four holes.Wherein two holes contrast as medicine, and 100TCID is attacked in two holes
50Virus, the 100ul/ hole.33 ℃ of 5%CO
2Cultivated 4-7 days, every day, observation of cell changed (CPE) to determine drug effect.When appearring in virus control, " ++ " then stop experiment.
2.3 experimental result: antiviral granule and antivirus oral liquid can suppress rhinovirus under non-toxic concn; Antiviral granule can suppress syncytial virus (RSV) under non-toxic concn.(the results are shown in Table 1-4)
Table 1 antiviral granule is to the inhibitory action of Influenza A1 virus blood clotting titre
Table 2 antiviral granule is to the inhibitory action of rhinovirus institute cytopathogenic effect (CPE)
Annotate: "
": dealed with medicine went, cell cavity, fragmentation occur, come off."-": cellular morphology is normal, and medicine can suppress virus; "+": medicine does not suppress virus, and CPE (cell fusion or circle contract) appears in cell
Table 3 antivirus oral liquid is to the inhibitory action of rhinovirus institute cytopathogenic effect (CPE)
Annotate: "
": dealed with medicine went, cell cavity, fragmentation occur, come off."-": cellular morphology is normal, and medicine can suppress virus; "+": medicine does not suppress virus, and CPE (cell fusion or circle contract) appears in cell
Table 4 antiviral granule is to the inhibitory action of RSV virus institute's cytopathogenic effect (CPE)
Annotate: "
": dealed with medicine went, cell cavity, fragmentation occur, come off."-": cellular morphology is normal, and medicine can suppress virus; "+": medicine does not suppress virus, and CPE (cell fusion or circle contract) appears in cell
Two, antiviral granule is to the dead protective effect of mice grippe pneumonia
1 experiment material
1.1 medicine: antiviral granule, press the preparation of embodiment 1 method.Antivirus oral liquid, available from the Guangzhou Xiangxue Pharmaceutical Co, lot number 200710010; The positive control medicine: virazole is provided lot number: 070804 by the Hubei institute of Pharmaceutical Industry.
1.2 animal: the NIH mice is provided by Guangdong Medical Lab Animal Center, meets a cleaning level experimental standard, lot number: the Guangdong word 2007A018 that checks and affirm, SCXK (Guangdong) 02004.The prescription that is rich in multiple composition that mouse feed provides for Traditional Chinese Medicine University Of Guangzhou's Experimental Animal Center.Feeding environment: room temperature is 23 ± 2 ℃, and relative humidity is 75 ± 10%.
1.3 seed culture of viruses: Influenza A1 virus FM1 strain, provided by Chinese medicine bioassay, the Mus lung adapts to after mice strengthens virulence, goes down to posterity three times in chick embryo allantoic cavity.And measure its median lethal dose(LD 50) LD
50
The medical clean work station of 1.4 equipment: YJ-875, Suzhou cleaning equipment company; Pressure steam sterilizer, Nanhua, Shanghai medical apparatus and instruments factory; Superpure water machine, Millipore; Ultra cold storage freezer, Sanyo Japan; Ten thousand/electronic analytical balance, FA/JA series, the flat instrument and meter company limited of last current chart; Ether, Tianjin chemical reagent company limited; 0.9% normal saline is opened and is helped Biology Pharmacy Co., Ltd; Table balance, Foochow balance equipment factory.
2 experimental techniques
2.1 dilution seed culture of viruses: before the test, get seed culture of viruses and be diluted to every 0.05 milliliter with physiological saline solution and contain 5 LD
50, put in the frozen water and preserve.
2.2 infecting mouse:, divide positive drug group (virazole), virus control group and trial drug group, male and female half and half the white mice random packet.Under the slight anesthesia of ether, with titration influenza virus collunarium infection, 4 of every Mus, about 0.05 milliliter.
3.3 administration: get 105 of NIH mices, male and female half and half are divided into 7 groups at random, and blank group, model group, virus control group, antivirus oral liquid group are for the high, medium and low dosage group of reagent.Each test group is all in virus attack beginning in preceding 3 days administration, every day 1 time.Continue administration behind the infective virus, every day 2 times, irritate 0.4ml/ administration of stomach.
3 observe: observe animal morbidity and death toll day by day, observed altogether 15 days.Dead mice anatomic observation Mus lung is observed the pulmonary consolidation degree that influenza infection causes.The degree that consolidation appears in lung is by following 5 grade standard records :-bilateral lung does not have pathological changes and occurs; +: pulmonary consolidation is less than 25% of whole lung surface area; ++: pulmonary consolidation accounts for 25～50% of whole lung surface area; +++: pulmonary consolidation accounts for 50～75% of whole lung surface area; ++ ++: pulmonary consolidation accounts for more than 75% of whole lung surface area.
4 calculate and statistical procedures: the result calculates dead protective rate and prolongs vital rates according to the observation:
Dead protective rate=[(matched group mortality rate-administration group mortality rate)/matched group mortality rate] * 100
Prolong vital rates=[(administration group on average survive number of days-matched group on average survive number of days)/matched group on average survive number of days] * 100
Adopt computer statistics software kit SPSS10.0 to carry out date processing, mortality rate X
2Check, the time-to-live is used rank test.
5 experimental results:
Table 5 antiviral granule is to the death protection of influenza virus infection in the mice body and the effect that prolongs disease Mus life
Annotate: * and virus control group be P＜0.05 relatively, * * P＜0.01
As can be seen from Table 5: the high, medium and low dosage group of antiviral granule is respectively the dead protective rate of first 1 type FM1 influenzae strain virus infected mice: 69.23%, 58.85%, 61.54%; Mean survival time was respectively 10.94 days, 10.23 days, 9.26 days.Isodose (under the 4.28g crude drug/kg), the antiviral granule agent to the dead protective rate of first 1 type FM1 influenzae strain virus infected mice and mice mean survival time than high respectively 16.16% and 1.12 day of antivirus oral liquid.The dead protective rate of positive control drug virazole group is: 84.62%, and mean survival time is 13.80 days.
Three, antiviral granule is to the inhibitory action of respiratory tract pathogenic bacterium
1 test material
1.1 experiment medicine
1.1.1 antivirus oral liquid: antivirus oral liquid is provided by the Guangzhou Xiangxue Pharmaceutical Co, lot number 20060426,0.45 gram crude drug/milliliters.
1.1.2 antiviral granule agent: press the preparation of embodiment 1 method, 5.76 gram crude drug/milliliters.
1.1.3 SHUANGHUANGLIAN KOUFUYE, He'nan Zhulin Zhongsheng Pharmaceutical Industry Co. Ltd produces.
1.1.4 hydrocortisone, lot number: C0117, Jin Jin pharmaceutical factory, Tianjin product.
1.1.5 Aspirin Enteric-coated Tablets, lot number: 060301,50 milligram/sheet, guilin pharmacy factory produces.
1.1.6 erythromycin lactobionate lot number: 20060602,30 ten thousand unit/bottles, Metro big pharmaceutical factory in Dalian produces.
1.2 laboratory animal
1.2.1 New Zealand white rabbit: provided by Guangdong Medical Lab Animal Center, body weight is 2.3～2.8kg, the laboratory animal quality certification number: 2006A025.
1.2.2SD rat: provided by Guangdong Medical Lab Animal Center, male, body weight is 220 ± 20g laboratory animal quality certification number: 2006A025.
1.2.3 Kunming mouse: provided by Guangdong Medical Lab Animal Center, male, body weight is 24 ± 2g laboratory animal quality certification number: 2006A023.
1.3 culture medium and reagent
1.3.1 nutrient broth: lot number: 061021, Nat'l Pharmaceutical ﹠ Biological Products Control Institute produces.
1.3.2 Nutrient agar: lot number: 070414, Nat'l Pharmaceutical ﹠ Biological Products Control Institute produces.
1.3.3 calf serum: lot number: 2006506, Science ﹠ Technology Center of Shanghai Second Emdical University tests two Room and produces.
1.3.4 sodium chloride (AR): lot number: 061201-3, Guangzhou Chemical Reagent Factory production.
1.3.5 anhydrous glucose (AR): lot number: 061120, consor thing Science and Technology Ltd. in source, Shanghai produces.
1.3.6 carrageenin: lot number: 127H1227, U.S. Sigma company produces.
1.3.7 dimethylbenzene: lot number: 061001-22, Guangzhou Chemical Reagent Factory production.
1.3.8 be dry yeast (bakery yeast): lot number: 005A, the sharp yeast company limited of Chinese-foreign joint venture prunus mume (sieb.) sieb.et zucc. mountain one horse product.
1.4 strain
Staphylococcus aureus (26003-21), escherichia coli (44103-), bacillus pyocyaneus (10104-1), streptococcus pneumoniae (31003-16), Hemolytic streptococcus (32210-17), hemophilus influenza (10105-32) are all purchased in Nat'l Pharmaceutical ﹠ Biological Products Control Institute.
1.5 key instrument
Water isolation type electrothermostat: model: PYX-DHS, scientific instrument related factory in Shanghai produces.
Portable High pressure steam sterilizer: model: YX-280, the bright Medical Equipment Plant in industrial group of Shanghai Medical Apparatus and Instruments Factory river produces.
GP2100 type electronic balance, German product.
1.6 experiment condition
The sterilizing room air purity reaches 10,000 grades, and superclean bench cleanliness factor 100 grades (parts) is up to specification.
Laboratory ventilation illumination is good, and temperature is 25～28 ℃, and humidity is 50～80%, the zoopery condition quality certification number: 2007C011.
2 methods and result
2.1 extracorporeal bacteria inhibitor test
2.1.1 the preparation of medicine:
All with sample stock solution as initial concentration, opposing by 1: 2 doubly is diluted to 10 Concentraton gradient, every pipe capacity is 1ml.
2.1.2 the preparation of culture medium
Broth bouillon: take by weighing nutrient broth, by 18 gram adding distil waters 1000 ml concns than preparation, after the heating for dissolving in 116 ℃ of autoclavings 20 minutes.
Agar culture medium: take by weighing Nutrient agar, by 34 gram adding distil waters 1000 ml concns than preparation, after the heating for dissolving in 116 ℃ of autoclavings 20 minutes.
The serum broth dextrose culture-medium: broth medium adds 10% calf serum and 1% glucose.
Defiber blood agar dextrose culture-medium: contain 1% glucose agar medium and in about 45 ℃, add 5% defiber blood, make blood plate and blood inclined-plane.
The phenol red broth bouillon of glucose: add the concentration ratio preparation of 5 gram glucoses and 6 milliliter of 0.4% phenol red aqueous solution by per 1000 milliliters of nutrient broths.
The phenol red broth bouillon of serum glucose: the phenol red broth bouillon of glucose adds 10% calf serum.
2.1.3 bacterium liquid preparation
Streptococcus pneumoniae, Hemolytic streptococcus are seeded in the blood agar slant medium respectively, staphylococcus aureus, escherichia coli, hemophilus influenza, bacillus pyocyaneus are seeded in agar slant culture-medium respectively, cultivated 20 hours for 37 ℃, get a platinum wire ear polyp tongue, be inoculated in respectively in serum broth or the broth bouillon, cultivated 20 hours for 37 ℃, bacteria suspension is diluted to final concentration 10 by 10 times of dilution methods
5CFU.
2.1.4 tube dilution method
This law can reflect the antibacterial ability that is tried thing, good reproducibility quantitatively, more sensitively.Antiviral granule agent and antivirus oral liquid, SHUANGHUANGLIAN KOUFUYE all with sample stock solution as initial concentration, phenol red meat soup of reuse glucose or the phenol red meat soup of serum glucose were opposed by 1: 2 and doubly are diluted to 10 Concentraton gradient; Erythromycin lactobionate is a starting point with 1000u/ml concentration, and opposing with phenol red meat soup of glucose or the phenol red meat soup of serum glucose doubly is diluted to 10 Concentraton gradient, every pipe 1ml; If positive control pipe (culture medium adds bacterium liquid), negative control pipe (culture medium adds medicinal liquid) and blank pipe (culture medium adding distil water), except that negative control pipe, blank pipe do not add the bacterium, all the other each pipes add above-mentioned dilution bacterium liquid 0.1ml, shake up, and put 37 ℃ and cultivate 20 hours.The result observes: with every arm respectively with the negative tube of its corresponding concentration relatively, whether according to culture tube clarity and change color, judging has bacteria growing.If culture tube presents clear and brightly, still present clear and bright person after the jolting, think this pipe asepsis growth, otherwise show that bacteria growing is arranged; Because metabolite of antibacterial and the phenol red generation metachromasia in the meat soup, its corresponding negative tube contrast if produce change color person's (deepening red or flavescence), is thought that this pipe has bacteria growing, otherwise is shown asepsis growth.The minimum drug dilution degree that antibacterial does not grow continues to be cultured to 48 hours for the minimum inhibitory concentration (MIC) of this medicine, if still present clear and bright asepsis growth state, this moment, minimum drug dilution degree was minimum bactericidal concentration (MBC).Each drug level is all established 2 parallel control tube, and experiment repeats 1 time, and MIC and MBC are the meansigma methods of 4 data.More than operation is all carried out under aseptic condition.
21.5 experimental result: see Table 6, table 7,
Agent of table 6. antiviral granule and antivirus oral liquid are to the minimum inhibitory concentration (MIC) of 6 kinds of common pathogens
Agent of table 7. antiviral granule and antivirus oral liquid are to the minimum bactericidal concentration (MBC) of 6 kinds of common pathogens
2.1.6 brief summary
Above result shows, under same experimental conditions, antiviral granule agent and antivirus oral liquid are to the common 6 kinds of pathogenic bacterium of upper respiratory tract infection, the sensitivity that each tool is different, wherein antivirus oral liquid is to the most responsive (MIC:0.03g/ml of staphylococcus aureus, MBC:0.10g/ml), secondly be bacillus pyocyaneus (MIC:0.16g/ml, MBC:0.222/ml); The most responsive (MIC is respectively: 0.04g/ml to staphylococcus aureus, bacillus pyocyaneus and hemophilus influenza in the antiviral granule agent, 0.08g/ml and 0.09g/ml, MBC is respectively: 0.035g/ml, 0.09g/ml, 0.16g/ml), next be streptococcus pneumoniae (MIC:0.14g/ml, MBC:0.27g/ml); Prompting thus, the antiviral granule agent totally presses down, bactericidal activity is stronger, secondly is antivirus oral liquid.
2.2 antiinflammatory is separated heat test in the body
2.2.1 on Carrageenan causes the influence of rat paw edema
Select healthy male rat for use, be divided into model control group, antivirus oral liquid group (4.9 gram crude drug/kilogram) and three dosage groups of antiviral granule agent (2.4,4.9 and 9.7 restrain crude drug/kilograms, are equivalent to 1 times, 2 times and 4 times of equivalents of the clinical feed ration of people respectively), positive control drug aspirin group (0.15 gram/kilogram) and SHUANGHUANGLIAN oral liquid group (10 milliliters/kilogram) at random by body weight.Administration group gastric infusion every day once, successive administration 3 days; Model control group gives the equal-volume distilled water.Respectively organize the right back sufficient normal volume of rat by capillary tube measurement by magnification method mensuration before causing inflammation.1h after the last administration, in the right back sufficient plantar subcutaneous injection 1% carrageenin 0.1ml of each group rat, measure the right back sufficient volume that causes scorching back 2h, 4h and 6h respectively, so that scorching forward and backward sufficient volume difference is the swelling degree, administration group and matched group relatively carry out statistical procedures (t check).See table 8 for details.
Each dosage group diagonal angle dish glue of table 8 antivirus oral liquid and antiviral granule agent cause rat paw edema influence (x ± SD, n=10)
Annotate: compare with model control group,
*P＜0.05,
*P＜0.01 (t check); Numeral is a suppression ratio in " () ".
Compare with antivirus oral liquid,
△P＜0.05 (q check);
The result shows, antivirus oral liquid and antiviral granule agent on Carrageenan cause rat paw edema all in various degree inhibitory action, wherein the overall antiphlogistic effects of antiviral granule agent is better, each dosage group all can obviously suppress to cause the pedal swelling of inflammation back 2h, 4h, 6h, compare with model control group, difference has significance meaning (P＜0.05 or 0.01); The overall antiphlogistic effects of antivirus oral liquid takes second place, and has only some dosage at some time point the foot swelling of inhibition effect to be arranged.
2.2.2 xylol causes the influence of mice auricle swelling
Select healthy male mice in kunming for use, be divided into model control group, antivirus oral liquid finished product group (7.0 gram crude drug/kilogram) and three dosage groups of antiviral granule agent (3.5,7.0 and 14.0 restrain crude drug/kilograms, are equivalent to 1 times, 2 times and 4 times of equivalents of the clinical feed ration of people respectively), positive control drug aspirin group (0.20 gram/kilogram) and SHUANGHUANGLIAN oral liquid group (20 milliliters/kilogram) at random by body weight.Administration group gastric infusion every day is (20 milliliters/kilogram) once, successive administration 5 days; Matched group waits the capacity distilled water.1h after the last administration is coated with that 0.03 milliliter of dimethylbenzene/only, left ear compares for the mouse right ear exterior feature.Taking off cervical vertebra after 15 minutes and put to death mice, with the card punch of diameter 8mm ears are downcut with the position homalographic, weigh with the precise electronic balance, is the swelling degree with the difference of left and right auricle weight.See table 9 for details.
Table 9 antivirus oral liquid and antiviral granule agent xylol cause mice auricle swelling influence (X ± SD, n=12)
Annotate: compare with model control group,
*P＜0.05,
*P＜0.01 (t check).
The result shows, antivirus oral liquid and antiviral granule agent xylol cause mice auricle swelling all in various degree inhibitory action, wherein three dosage groups of antiviral granule agent all can obviously suppress auricle edema, compare with model control group, and difference has significance meaning (P＜0.05 or 0.01).
2.2.3 to the swollen influence that forms of rat granuloma
Healthy rat is selected in experiment for use, be divided into model control group, antivirus oral liquid group (9.7 gram crude drug/kilogram) and three dosage groups of antiviral granule agent (4.9,9.7,19.5 restrain crude drug/kilograms, are equivalent to 2 times, 4 times and 8 times of equivalents of the clinical feed ration of people respectively), positive control drug hydrocortisone group (20 milligrams/kilogram) and SHUANGHUANGLIAN oral liquid group (20 milliliters/kilogram) at random by body weight.Rat after the routine sterilization, at abdominal scissors one osculum, is expanded subcutaneous tissue with vascular forceps under pentobarbital sodium anesthesia, (weight is that 10 ± 1mg) implantation both sides groin are subcutaneous to the cotton balls of will sterilizing, and then sews up.Cotton balls adds penicillin 0.1mg, in case infect.Second day after operation begins gastric infusion, once a day, successive administration 7 days, animal is put to death in dislocation in the 8th day, peels off and take out granuloma induced by implantation of cotton pellets, rejects fatty tissue, weighs after 1 hour in 70 ℃ of oven dryings, deducts cotton balls weight, is granuloma weight.Granuloma weight is represented with milligram/100 gram body weight, be the results are shown in Table 10.
Table 10 antivirus oral liquid and antiviral granule agent are to the swollen influence that forms of rat granuloma (X ± SD)
Annotate: compare with model control group:
*P＜0.05,
*P＜0.01 (t check).
The result shows, each dosage group of antivirus oral liquid and antiviral granule agent is to the swollen inhibitory action that all has in various degree of rat granuloma, wherein the basic, normal, high dosage of antiviral granule agent all can obviously suppress the hypertrophy of granulation tissue, suppression ratio is respectively 65%, 53% and 61%, compares P＜0.01 with model control group.
2.2.4 dry yeast is caused the influence of rat fever
Select healthy male rat for use, survey body temperature 2 times with electronic clinical thermometer every day before the administration in anus, continuous 2 days, stimulates so that rat adapts to operation.Choose the rat that body temperature fluctuation is no more than 0.3 ℃ and be divided into 16 groups: three dosage groups (2.4,4.9 and 9.7 restrain crude drug/kilograms, are equivalent to 1 times, 2 times and 4 times of equivalents of the clinical consumption per day of people respectively), positive control drug aspirin group (0.15 gram/kilogram) and the SHUANGHUANGLIAN oral liquid group (10 milliliters/kilogram) of model control group, antivirus oral liquid group (4.9 gram crude drug/kilogram) and antiviral granule agent.Administration group gastric infusion every day once, successive administration 3 days; Model control group gives the equal-volume distilled water.Per hour measure rat temperature 1 time before the experiment administration on the same day, continuous 2 times, get average as normal body temperature.Immediately in the sterilised yeast suspension 10ml/kg of nape portion subcutaneous injection 20%, bring out rat fever after each Mus administration, measure 4,6,8 hours rat anus temperature after the pyrogenicity then respectively.The results are shown in Table 11.
Each dosage group of table 11 antivirus oral liquid and antiviral granule agent to dry yeast cause the influence that rat temperature raises (X ± SD, n=10)
Annotate: compare with model control group,
*P＜0.05,
*P＜0.01 (t check).Numeral is a suppression ratio in " () "
The result shows, each dosage group of antivirus oral liquid and antiviral granule agent causes rat temperature to dry yeast and raises in various degree refrigeration function is arranged, wherein the antiviral granule agent totally to separate thermal effect better, each dosage to rat pyrogenicity after the fervescence of 4h, 6h, 8h the trivial solution heat effect is arranged, compare with model control group, difference has significance meaning (P＜0.05 or 0.01).
The specific embodiment
The preparation of example 1 granule
1, extract: get Herba Pogostemonis 2.9Kg, Rhizoma Acori Graminei 2.5Kg, in the extraction kettle of packing into, in 55 ℃ of pressure 28MPa, temperature extraction 2 hours down, collect volatile oil 198g, medicinal residues are standby.
Medicinal residues and Fructus Forsythiae 4.6Kg, Radix Isatidis 12.9Kg, Radix Curcumae 2.5Kg, Radix Rehmanniae 3.2Kg add 8 times of amount 80% ethanol, heating and refluxing extraction 2 times, and each 2 hours, emit extracting solution through horizontal screw unloading filter centrifugal machine, merge extracted twice liquid, standby.
Get Rhizoma Anemarrhenae 2.5Kg, Gypsum Fibrosum 5.7Kg, Rhizoma Phragmitis 6.1Kg coarse powder, the water that adds 8 times of amounts extracts 2 times down at 80 ℃, and each 2 hours, emit extracting solution through horizontal screw unloading filter centrifugal machine, merge extracted twice liquid, standby.
2, concentrate: with water extract and alcohol extract is 100 ℃ and 60 ℃ of vacuum for concentrating under reduced pressure Cheng Shui under the-0.06MPa carries with the about 11800mL of alcohol extraction concentrated solution total (50 ℃ time survey its density be 1.12g/mL) in temperature respectively, measures its paste-forming rate simultaneously, standby.
3, drying: the water of step 1 carried with the alcohol extraction concentrated solution merge, add the dextrin that is equivalent to its dried cream amount 30%, the dissolving mixing, obtaining extractum 12000mL (50 ℃ time survey its density be 1.15g/mL), is that 155～165 ℃, air outlet temperature are that 100 ℃, pressure are to carry out spray drying under the 1bar condition promptly to get 9.27Kg yellowish-brown dried powder at intake air temperature.
4, the preparation of clathrate: get step 1 gained volatile oil and dilute with dehydrated alcohol in 1: 1 ratio, splash in the beta-schardinger dextrin-saturated solution of 6 times of amount volatile oil weight, stirred 2 hours in 50 ℃, mixing speed is 200rpm, left standstill 24 hours, sucking filtration, the control temperature promptly gets 1.39Kg white powder clathrate carrying out vacuum drying below 60 ℃, and inclusion rate is 70.2%.
5, granulate: with the yellowish-brown dried powder of step 3 gained and the dextrin mixing of step 4 gained volatile oil clathrate compound and 1.5 times of amounts of dry powder weight, add 30% 95% ethanol moistening, the system soft material sieves the 40 ℃ of heat-wind circulate dryings of granule that sieve, granulate.
6, fill, sterilization: the dried particles fill becomes the 2.5g/ bag, and sterilization promptly.The consumption of said preparation is every day twice, and each 1～2 bag, warm water is taken after mixing it with water.
Detect with high performance liquid chromatography, record the content that contains chimonin and phillyrin in the antiviral granule agent and be respectively 0.608mg/g, 0.124mg/g; Detect with gas chromatography, recording patchouli alcohol content is every bag of 51 μ g.
The preparation of example 2 granules
1, extract: get Herba Pogostemonis 2.9Kg, Rhizoma Acori Graminei 2.5Kg, in the extraction kettle of packing into, in 55 ℃ of pressure 28MPa, temperature extraction 2 hours down, collect volatile oil 203g, medicinal residues are standby.
Medicinal residues and Fructus Forsythiae 4.6Kg, Radix Isatidis 12.9Kg, Radix Curcumae 2.5Kg, Radix Rehmanniae 3.2Kg add 10 times of amount 80% ethanol, heating and refluxing extraction 1 time, and each 3 hours, emit extracting solution through horizontal screw unloading filter centrifugal machine, merge, get alcohol extract 56000ml, standby.
Get Rhizoma Anemarrhenae 2.5Kg, Gypsum Fibrosum 5.7Kg, Rhizoma Phragmitis 6.1Kg coarse powder, the water that adds 10 times of amounts extracts 1 time down at 80 ℃, and each 1 hour, emit extracting solution through horizontal screw unloading filter centrifugal machine, merge, get water extract 28000ml, standby.
2, concentrate: operation is with embodiment one, water is carried with the alcohol extraction concentrated solution and is added up to about 11800mL (50 ℃ time survey its density be 1.12g/mL), standby.
3, drying: operation is with embodiment one, obtaining extractum 12000mL (50 ℃ time survey its density be 1.15g/mL), is that 160～170 ℃, air outlet temperature are that 110 ℃, pressure are to carry out spray drying under the 1.4bar condition promptly to get 9.25Kg yellowish-brown dried powder at intake air temperature.
4, the preparation of clathrate: get step 1 gained volatile oil and dilute with dehydrated alcohol in 1: 1 ratio, splash in the beta-schardinger dextrin-saturated solution of 6 times of amount volatile oil weight, stirred 1 hour in 40 ℃, mixing speed is 200rpm, left standstill 24 hours, sucking filtration, the control temperature promptly gets 1.36Kg white powder clathrate carrying out vacuum drying below 60 ℃, and inclusion rate is 64.9%.
5, granulate: with the yellowish-brown dried powder of step 3 gained and the dextrin mixing of step 4 gained volatile oil clathrate compound and 1.5 times of amounts of dry powder weight, add 30% 95% ethanol moistening, the system soft material sieves the 40 ℃ of heat-wind circulate dryings of granule that sieve, granulate.
6, fill, sterilization: the dried particles fill becomes the 2.5g/ bag, and sterilization promptly.The consumption of said preparation is every day twice, and each 1～2 bag, warm water is taken after mixing it with water.
Detect with high performance liquid chromatography, record the content that contains chimonin and phillyrin in the antiviral granule agent and be respectively 0.596mg/g, 0.118mg/g; Detect with gas chromatography, recording patchouli alcohol content is every bag of 49 μ g.
The preparation of example 3 granules
1, extract: operation is collected volatile oil 202g with embodiment one, water extract and alcohol extract each 28000,56000mL, standby.
2, concentrate: operation is with embodiment one, water is carried with the alcohol extraction concentrated solution and is added up to about 11900mL (50 ℃ time survey its density be 1.11g/mL), standby.
3, drying: operation is with embodiment one, obtaining extractum 12000mL (50 ℃ time survey its density be 1.15g/mL), is that 165～175 ℃, air outlet temperature are that 110 ℃, pressure are to carry out spray drying under the 1.2bar condition promptly to get 9.27Kg yellowish-brown dried powder at intake air temperature.
4, the preparation of clathrate: get step 1 gained volatile oil and dilute with dehydrated alcohol in 1: 1 ratio, splash in the beta-schardinger dextrin-saturated solution of 6 times of amount volatile oil weight, stirred 3 hours in 60 ℃, mixing speed is 300rpm, left standstill 24 hours, sucking filtration, the control temperature promptly gets 1.38Kg white powder clathrate carrying out vacuum drying below 60 ℃, and inclusion rate is 67.8%.
5, granulate: with yellowish-brown dried powder and the volatile oil clathrate compound of step 4 gained and the dextrin mixing of 1.5 times of amounts of dry powder weight of step 3 gained, add 30% 95% ethanol moistening, the system soft material sieves the 40 ℃ of heat-wind circulate dryings of granule that sieve, granulate.
6, fill, sterilization: the dried particles fill becomes the 2.5g/ bag, and sterilization promptly.The consumption of said preparation is every day twice, and each 1～2 bag, warm water is taken after mixing it with water.
Detect with high performance liquid chromatography, record the content that contains chimonin and phillyrin in the antiviral granule agent and be respectively 0.602mg/g, 0.120mg/g; Detect with gas chromatography, recording patchouli alcohol content is every bag of 53 μ g.
The preparation of example 4 granules
1, extract: operation is collected volatile oil 196g with embodiment one, water extract and alcohol extract each 28000,56000mL, standby.
2, concentrate: operation is with embodiment one, water is carried with the alcohol extraction concentrated solution and is added up to about 11530mL (50 ℃ time survey its density be 1.12g/mL), standby.
3, drying: operation is with embodiment one, obtaining extractum 11600mL (50 ℃ time survey its density be 1.14g/mL), is that 165～175 ℃, air outlet temperature are that 90 ℃, pressure are to carry out spray drying under the 1.4bar condition promptly to get 9.21Kg yellowish-brown dried powder at intake air temperature.
4, the preparation of clathrate: get step 1 gained volatile oil and dilute with dehydrated alcohol in 1: 1 ratio, splash in the beta-schardinger dextrin-saturated solution of 8 times of amount volatile oil weight, stirred 1 hour in 50 ℃, mixing speed is 300rpm, left standstill 24 hours, sucking filtration, the control temperature promptly gets 1.42Kg white powder clathrate carrying out vacuum drying below 60 ℃, and inclusion rate is 72.9%.
5, granulate: with yellowish-brown dried powder and the volatile oil clathrate compound of step 4 gained and the dextrin mixing of 1.5 times of amounts of dry powder weight of step 3 gained, add 30% 95% ethanol moistening, the system soft material sieves the 40 ℃ of heat-wind circulate dryings of granule that sieve, granulate.
6, fill, sterilization: the dried particles fill becomes the 2.5g/ bag, and sterilization promptly.The consumption of said preparation is every day twice, and each 1～2 bag, warm water is taken after mixing it with water.
Detect with high performance liquid chromatography, record the content that contains chimonin and phillyrin in the antiviral granule agent and be respectively 0.594mg/g, 0.116mg/g; Detect with gas chromatography, recording patchouli alcohol content is every bag of 48 μ g.
The preparation of example 5 granules
1, extract: operation is collected volatile oil 201g with embodiment one, water extract and alcohol extract each 28000,56000mL, standby.
2, concentrate: operation is with embodiment one, water is carried with the alcohol extraction concentrated solution and is added up to about 11860mL (50 ℃ time survey its density be 1.12g/mL), standby.
3, drying: operation is with embodiment one, obtaining extractum 12000mL (50 ℃ time survey its density be 1.15g/mL), is that 175～185 ℃, air outlet temperature are that 110 ℃, pressure are to carry out spray drying under the 1.4bar condition promptly to get 9.20Kg yellowish-brown dried powder at intake air temperature.
4, the preparation of clathrate: get step 1 gained volatile oil and dilute with dehydrated alcohol in 1: 1 ratio, splash in the beta-schardinger dextrin-saturated solution of 8 times of amount volatile oil weight, stirred 2 hours in 60 ℃, mixing speed is 300rpm, left standstill 24 hours, sucking filtration, the control temperature promptly gets 1.43Kg white powder clathrate carrying out vacuum drying below 60 ℃, and inclusion rate is 74.3%.
5, granulate: with yellowish-brown dried powder and the volatile oil clathrate compound of step 4 gained and the dextrin mixing of 1.5 times of amounts of dry powder weight of step 3 gained, add 30% 95% ethanol moistening, the system soft material sieves the 40 ℃ of heat-wind circulate dryings of granule that sieve, granulate.
6, fill, sterilization: the dried particles fill becomes the 2.5g/ bag, and sterilization promptly.The consumption of said preparation is every day twice, and each 1～2 bag, warm water is taken after mixing it with water.
Detect with high performance liquid chromatography, record the content that contains chimonin and phillyrin in the antiviral granule agent and be respectively 0.612mg/g, 0.122mg/g; Detect with gas chromatography, recording patchouli alcohol content is every bag of 52 μ g.
The preparation of example 6 granules
1, extract: operation is collected volatile oil 199g with embodiment one, water extract and alcohol extract each 28000,56000mL, standby.
2, concentrate: operation is with embodiment one, water is carried with the alcohol extraction concentrated solution and is added up to about 11800mL (50 ℃ time survey its density be 1.12g/mL), standby.
3, drying: operation is with embodiment one, obtaining extractum 12000mL (50 ℃ time survey its density be 1.15g/mL), is that 165～175 ℃, air outlet temperature are that 110 ℃, pressure are to carry out spray drying under the 1.2bar condition promptly to get 9.22Kg yellowish-brown dried powder at intake air temperature.
4, the preparation of clathrate: get step 1 gained volatile oil and dilute with dehydrated alcohol in 1: 1 ratio, splash in the beta-schardinger dextrin-saturated solution of 8 times of amount volatile oil weight, stirred 1 hour in 40 ℃, mixing speed is 300rpm, left standstill 24 hours, sucking filtration, the control temperature promptly gets 1.42Kg white powder clathrate carrying out vacuum drying below 60 ℃, and inclusion rate is 72.1%.
5, granulate: with the yellowish-brown dried powder of step 3 gained and the dextrin mixing of step 4 volatile oil clathrate compound and 1.5 times of amounts of dry powder weight, add 30% 95% ethanol moistening, the system soft material sieves the 40 ℃ of heat-wind circulate dryings of granule that sieve, granulate.
6, fill, sterilization: the dried particles fill becomes the 2.5g/ bag, and sterilization promptly.The consumption of said preparation is every day twice, and each 1～2 bag, warm water is taken after mixing it with water.
Detect with high performance liquid chromatography, record the content that contains chimonin and phillyrin in the antiviral granule agent and be respectively 0.602mg/g, 0.123mg/g; Detect with gas chromatography, recording patchouli alcohol content is every bag of 50 μ g.
The preparation of example 7 granules
1, extract: operation is collected volatile oil 204g with embodiment one, water extract and alcohol extract each 28000,56000mL, standby.
2, concentrate: operation is with embodiment one, water is carried with the alcohol extraction concentrated solution and is added up to about 11800mL (50 ℃ time survey its density be 1.12g/mL), standby.
3, drying: operation is with embodiment one, obtaining extractum 12000mL (50 ℃ time survey its density be 1.15g/mL), is that 165～175 ℃, air outlet temperature are that 110 ℃, pressure are to carry out spray drying under the 1.2bar condition promptly to get 9.21Kg yellowish-brown dried powder at intake air temperature.
4, the preparation of clathrate: get step 1 gained volatile oil and dilute with dehydrated alcohol in 1: 1 ratio, splash in the beta-schardinger dextrin-saturated solution of 10 times of amount volatile oil weight, stirred 1 hour in 60 ℃, mixing speed is 300rpm, left standstill 24 hours, sucking filtration, the control temperature promptly gets 1.44Kg white powder clathrate carrying out vacuum drying below 60 ℃, and inclusion rate is 75.8%.
5, granulate: with the yellowish-brown dried powder of step 3 gained and the dextrin mixing of step 4 volatile oil clathrate compound and 1.5 times of amounts of dry powder weight, add 30% 95% ethanol moistening, the system soft material sieves the 40 ℃ of heat-wind circulate dryings of granule that sieve, granulate.
6, fill, sterilization: the dried particles fill becomes the 2.5g/ bag, and sterilization promptly.The consumption of said preparation is every day twice, and each 1～2 bag, warm water is taken after mixing it with water.
Detect with high performance liquid chromatography, record that the content of chimonin and phillyrin is respectively 0.593mg/g, 0.116mg/g in the antiviral granule agent; Detect with gas chromatography, recording patchouli alcohol content is every bag of 49 μ g.
The preparation of example 8 granules
1, extract: operation is collected volatile oil 203g with embodiment one, water extract and alcohol extract each 28000,56000mL, standby.
2, concentrate: operation is with embodiment one, water is carried with the alcohol extraction concentrated solution and is added up to about 11800mL (50 ℃ time survey its density be 1.12g/mL), standby.
3, drying: operation is with embodiment one, obtaining extractum 12000mL (50 ℃ time survey its density be 1.15g/mL), is that 175～185 ℃, air outlet temperature are that 100 ℃, pressure are to carry out spray drying under the 1.4bar condition promptly to get 9.26Kg yellowish-brown dried powder at intake air temperature.
4, the preparation of clathrate: get step 1 gained volatile oil and dilute with dehydrated alcohol in 1: 1 ratio, splash in the beta-schardinger dextrin-saturated solution of 10 times of amount volatile oil weight, stirred 2 hours in 40 ℃, mixing speed is 300rpm, left standstill 24 hours, sucking filtration, the control temperature promptly gets 1.45Kg white powder clathrate carrying out vacuum drying below 60 ℃, and inclusion rate is 76.4%.
5, granulate: with the yellowish-brown dried powder of step 3 gained and the dextrin mixing of step 4 volatile oil clathrate compound and 1.5 times of amounts of dry powder weight, add 30% 95% ethanol moistening, the system soft material sieves the 40 ℃ of heat-wind circulate dryings of granule that sieve, granulate.
6, fill, sterilization: the dried particles fill becomes the 2.5g/ bag, and sterilization promptly.The consumption of said preparation is every day twice, and each 1～2 bag, warm water is taken after mixing it with water.
Detect with high performance liquid chromatography, record the content that contains chimonin and phillyrin in the antiviral granule agent and be respectively 0.606mg/g, 0.126mg/g; Detect with gas chromatography, recording patchouli alcohol content is every bag of 53 μ g.
The preparation of example 9 granules
1, extract: operation is collected volatile oil 203g with embodiment one, water extract and alcohol extract each 28000,56000mL, standby.
2, concentrate: operation is with embodiment one, water is carried with the alcohol extraction concentrated solution and is added up to about 11850mL (50 ℃ time survey its density be 1.13g/mL), standby.
3, drying: operation is with embodiment one, obtaining extractum 12000mL (50 ℃ time survey its density be 1.15g/mL), is that 155～165 ℃, air outlet temperature are that 110 ℃, pressure are to carry out spray drying under the 1.2bar condition promptly to get 9.22Kg yellowish-brown dried powder at intake air temperature.
4, the preparation of clathrate: get step 1 gained volatile oil and dilute with dehydrated alcohol in 1: 1 ratio, splash in the beta-schardinger dextrin-saturated solution of 10 times of amount volatile oil weight, stirred 3 hours in 50 ℃, mixing speed is 300rpm, left standstill 24 hours, sucking filtration, the control temperature promptly gets 1.44Kg white powder clathrate carrying out vacuum drying below 60 ℃, and inclusion rate is 76.2%.
5, granulate: with the yellowish-brown dried powder of step 3 gained and the dextrin mixing of step 4 gained volatile oil clathrate compound and 1.5 times of amounts of dry powder weight, add 30% 95% ethanol moistening, the system soft material sieves the 40 ℃ of heat-wind circulate dryings of granule that sieve, granulate.
6, fill, sterilization: the dried particles fill becomes the 2.5g/ bag, and sterilization promptly.The consumption of said preparation is every day twice, and each 1～2 bag, warm water is taken after mixing it with water.
Detect with high performance liquid chromatography, record the content that contains chimonin and phillyrin in the antiviral granule agent and be respectively 0.607mg/g, 0.114mg/g; Detect with gas chromatography, recording patchouli alcohol content is every bag of 51 μ g.
The preparation of example 10 oral liquids
1, extract: operation is collected volatile oil 203g with embodiment one, water extract and alcohol extract each 28000,56000mL, standby.
2, concentrate: operation is with embodiment one, water is carried with the alcohol extraction concentrated solution and is added up to about 11850mL (50 ℃ time survey its density be 1.13g/mL), standby.
3, the preparation of clathrate: get step 1 gained volatile oil and dilute with dehydrated alcohol in 1: 1 ratio, splash in the beta-schardinger dextrin-saturated solution of 10 times of amount volatile oil weight, in 50 ℃ of stirrings 3 hours, mixing speed was 300rpm, obtain beta-cyclodextrin inclusion compound liquid 4410g, inclusion rate is 76.2%.
4, join filter: after the antiviral concentrated solution 11850mL filtration with step 2 gained, add Mel 4200g, sucrose 5600g beta-cyclodextrin inclusion compound liquid 4410g mixing, be filtered to clarification, add water and be settled to 100L, standardize solution liquid filters.
5, fill, sterilization: the fill of standardize solution liquid becomes 10mL/ to prop up, and sterilization promptly.The consumption of said preparation is every day three times, and is each 1～2, oral.
Detect with high performance liquid chromatography, record that the content of chimonin and phillyrin is respectively 2.86mg/10mL, 0.582mg/10mL in the antivirus oral liquid of the present invention; Detect with gas chromatography, the content that records patchouli alcohol is 49 μ g/10ml.
Example 11 Comparative Examples: the antivirus oral liquid of former prepared
1, extracts: by the prescription and the technology of former antivirus oral liquid, get Radix Isatidis 900g, Gypsum Fibrosum 400g, Rhizoma Phragmitis 425g, Radix Rehmanniae 225g, Radix Curcumae 175g, Rhizoma Anemarrhenae 175g, Rhizoma Acori Graminei 175g, Herba Pogostemonis 200g, Fructus Forsythiae 325g, the water that adds 8 times of amounts, the interlayer heated and boiled was extracted 3 hours, collect the emulsion 1960mL that contains volatile oil simultaneously, standby.Extracting liquid filtering separates, and is standby.
Medicinal residues add the water extraction 1 hour 30 minutes of 6 times of amounts again, and extracting liquid filtering separates, with the first time extracting solution merge, standby.Medicinal residues discard.
2, concentrate: merge extractive liquid,, at vacuum concentration below 65 ℃ to relative density 1.12, put and be chilled to room temperature, in stirring, slowly add 85% ethanol, make to contain concentration of alcohol and reach 70 ± 2%, left standstill 24 hours, filter, getting supernatant recovery ethanol and being concentrated into relative density is that 1.09 (surveying for 20 ℃) get antiviral concentrated solution total 2100mL, standby.
3, join filter: after the antiviral concentrated solution 11850mL filtration with step 2 gained, add Mel 4200g, sucrose 5600g beta-cyclodextrin inclusion compound liquid 4410g mixing, be filtered to clarification, add water and be settled to 100L, standardize solution liquid filters.
4, fill, sterilization: the fill of standardize solution liquid becomes 10mL/ to prop up, and sterilization promptly.The consumption of said preparation is every day three times, and is each 1～2, oral.
Detect with high performance liquid chromatography, record that the content of chimonin and phillyrin is respectively 0.183mg/10mL, 0.308mg/10mL in the antiviral granule agent of former prepared; Detect with gas chromatography, the content that records patchouli alcohol is 36 μ g/10ml.
Claims (2)
1, a kind of preparation method of antiviral composition, this method comprises following step:
(1) get Herba Pogostemonis 2.9 weight portions, Rhizoma Acori Graminei coarse powder 2.5 weight portions, CO packs into
2In the supercritical extraction reactor, extracted 2 hours down for 55 ℃, collect volatile oil, stay medicinal residues standby in pressure 28MPa, temperature;
(2) get the Rhizoma Anemarrhenae 2.5 weight portions, Gypsum Fibrosum 5.7 weight portions, Rhizoma Phragmitis coarse powder 6.1 weight portions, add 8～10 times in water, 80 ℃ are extracted 2～3 times down, each 1～2 hour, filter, merge extractive liquid,, temperature be 100 ℃ of vacuums be-0.08～-0.04MPa under concentrating under reduced pressure become clear paste, making its density under 50 ℃ is 1.05～1.15g/mL;
(3) getting Fructus Forsythiae 4.6 weight portions, Radix Isatidis 12.9 weight portions, Radix Curcumae 2.5 weight portions, Radix Rehmanniae 3.2 weight portions and step (1) gained medicinal residues mixes, extract 2～3 times with 8～10 times of 80% alcohol heating reflux, each 1～2 hour, filter, merge extractive liquid,, with ethanol extract temperature be 60 ℃ of vacuums be-0.08～-0.04MPa under concentrating under reduced pressure become clear paste, making its density under 50 ℃ is 1.05～1.15g/mL;
(4) clear paste of getting volatile oil, step (2) and (3) gained of step (1) gained is prepared into preparation commonly used according to a conventional method.
2, preparation method according to claim 1, wherein the described volatile oil of step (4) is before being used to prepare preparation, the following processing of process: with equal-volume dehydrated alcohol dilution doubly, splash into then in the beta-schardinger dextrin-saturated solution of 6～10 times of amount volatile oil weight, in 40～60 ℃ of stirrings 3～5 hours, left standstill 24 hours.
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| CN200910193045A CN101670074A (en) | 2009-10-13 | 2009-10-13 | Preparation method for antivirus compound |
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| CN200910193045A CN101670074A (en) | 2009-10-13 | 2009-10-13 | Preparation method for antivirus compound |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105362283A (en) * | 2014-08-07 | 2016-03-02 | 富力 | Applications of phillyrin/phillygenin composition in preparation of drugs or health products for relieving or/and treatment of viral diseases |
| CN109172780A (en) * | 2018-11-20 | 2019-01-11 | 邹敬韬 | A kind of preparation method of compound Moschus injection |
| CN109999162A (en) * | 2019-05-15 | 2019-07-12 | 安徽东盛友邦制药有限公司 | A kind of preparation method of the means of supercritical extraction effective component of antiviral oral liquor |
-
2009
- 2009-10-13 CN CN200910193045A patent/CN101670074A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105362283A (en) * | 2014-08-07 | 2016-03-02 | 富力 | Applications of phillyrin/phillygenin composition in preparation of drugs or health products for relieving or/and treatment of viral diseases |
| CN105362283B (en) * | 2014-08-07 | 2018-07-27 | 富力 | Forsythin/phillygenol composition is preparing the application in alleviating or/and treating the drug or health products of viral disease |
| CN109172780A (en) * | 2018-11-20 | 2019-01-11 | 邹敬韬 | A kind of preparation method of compound Moschus injection |
| CN109999162A (en) * | 2019-05-15 | 2019-07-12 | 安徽东盛友邦制药有限公司 | A kind of preparation method of the means of supercritical extraction effective component of antiviral oral liquor |
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