CN100448450C - Pharyngolaryngitis-treating pharmaceutical compositions and its preparing method - Google Patents

Abstract

The present invention provides a medicinal composition for treating pharyngolaryngitis, which is a medicinal preparation prepared from active components and pharmaceutically acceptable auxiliary materials or auxiliary components, wherein the active components are iristectorum maxim and extract of total flavonoid, and the percentage content of the extract of total flavonoid by tectoridin C22H22O11 is equal to or more than 55%. The present invention is characterized in that the quality of the medicinal composition is stable and controllable, the medicinal effect components of tectoridin, iristectorin A, tectoridin aglycon and iridin aglycon in the medicinal composition are determined, and the medicinal composition performs a synergic effective function through the compounding of the 4 components. The medicinal composition can obviously inhibit acute laryngopharyngitis and upper respiratory tract infection. The medicinal composition takes effect and relieves symptoms rapidly, a patient with hoarse voice caused by swelling and pain in throat can return to normal after taking the medicinal composition for several hours, and swelling and pain in throat is obviously alleviated or eliminated within a plurality of hours; if the patient continuously takes the medicinal composition for 3 to 7 days, the patient can be healed. The medicinal composition is a fast-effective medicine with curative effect and has the advantages of low cost and high yield; the medicinal composition provides a new medicinal choice for clinical operation.

Description

The purposes of Rhizoma Iridis Tectori extractive of general flavone

Technical field

The present invention relates to the purposes of Rhizoma Iridis Tectori extractive of general flavone, belong to the field of Chinese medicines.

Background technology

The traditional Chinese medical science thinks that acute pharyngolaryngitis belongs to wind-heat evil toxin attacks outward, and pathogenic heat be jammed sore throat (acute pharyngitis), acute hoarseness (acute laryngitis) due to the lungs are to have a strong impact on healthy commonly encountered diseases, frequently-occurring disease, often is accompanied by flu and produces.Onset is anxious, is cardinal symptom with laryngopharynx swelling and pain, hoarseness, headache fever, cough expectorant Huang, and further throat, the hyperemia of nasal mucosa edema can appear in aggravation.Modern medicine confirms: acute pharyngolaryngitis mostly is due to the virus, mainly contains influenza virus, parainfluenza virus, adenovirus, Coxsackie virus, respiratory syncytial virus, rhinovirus etc., and antibiotics is invalid to this type of viral infection.Clinical treatment acute pharyngolaryngitis and this type of upper respiratory tract infection Western medicine commonly used has amantadine, rimantadine, ribavirin (virazole), moroxydine (moroxydine) etc.They have sure effect to some virus that causes acute pharyngolaryngitis, but curative effect is comparatively single, there are not antibiotic antiinflammatory, antipyretic-antalgic, antitussive action, promptly undesirable to the controlling symptoms effect, and the equal more serious toxic and side effects of tool, as cause anemia, platelet and leukopenia, hematuria, dizziness, ataxia, edema, visual loss, gastrointestinal reaction, immunosuppressant, diarrhoea and renal function injury etc.The conventional Chinese medicine of treatment acute pharyngolaryngitis and upper respiratory tract infection has the compound preparation of creat formulation series, Herba Houttuyniae preparation series, Radix Isatidis preparation series, 'Shuang Hualian ' series and other heat-clearing and detoxifying herb.The characteristics of these Chinese medicine preparation are: not only virus is had certain inhibitory action, and have antiinflammatory, effect such as the analgesia of bringing down a fever, but antivirus action is remarkable inadequately, mostly be compound preparation, at concrete indication have nothing in common with each other, the quality instability, contained effective site or effective ingredient are indeterminate, and take, carry, preserve equal inconvenience.

Rhizoma Iridis Tectori is an irides Rhizoma Iridis Tectori Iris tectorum Maxim. rhizome.Have removing food stagnancy, removing blood stasis with potent drugs, Hang Shui, antidotal effect.Treatment dyspepsia distension ， WEIJIA is gathered expansion, toxic swelling, hemorrhoid complicated by anal fistula, traumatic injury.Application number: 200410022293.X discloses the Rhizoma Iridis Tectori water extract, and in preparation treatment stomatitis, pharyngolaryngitis and chronic bronchitis and relieve the effect of alcohol, the purposes in the slimming medicine.The disclosure bibliographical information be the crude extract of Rhizoma Iridis Tectori, indication is many, effective ingredient is indeterminate.

Summary of the invention

Technical scheme of the present invention provides the purposes of irides Rhizoma Iridis Tectori Iris tectorum Maxim. extractive of general flavone in the preparation antiviral drugs, and wherein, the percentage composition of total flavones is with iridin C in the extractive of general flavone ₂₂H ₂₂O ₁₁Calculate 〉=55%.

Further, contain in the above-mentioned total flavones: iridin tectoridin, iristectorin A iristectorinA, iris aglycone tectorigenin, wild flag aglycon irigenin

Further, the weight proportion iridin of four isoflavones components in the total flavones: iristectorin A: iris aglycone: the wild flag aglycon is 1: 0.1～0.4: 0.15～0.6: 0.03～0.1.

Further, in the total flavones content of four isoflavones components than iridin: iristectorin A: iris aglycone: the wild flag aglycon is 1: 0.18: 0.29: 0.056.

Described Rhizoma Iridis Tectori total flavones is to extract preparation by following method:

A, get irides Rhizoma Iridis Tectori (Iris tectorum Maxim.) medical material, pulverize, add 70% alcohol reflux, concentrate, extractum;

In b, the extractum dissolving entry with a step, filter, filtrate is passed through macroporous adsorptive resins;

C, wash the fat post with water, effluent discards, and when effluent is nearly achromatism and clarity solution, puts solid carbon dioxide, and with 75～90% ethanol elutions, eluent filters, and concentrates and promptly gets total flavones of the present invention.

Wherein, the weight proportion of the amount of resin of the described macroporous adsorptive resins of b step and primary crude drug is in the extracting method of described Rhizoma Iridis Tectori total flavones: 3: 1.

Described preparation is: capsule, granule, tablet, pill, oral liquid.

The present invention also provides the preparation method of this antiviral drugs, comprises the steps:

A, get irides Rhizoma Iridis Tectori Iris tectorum Maxim. medical material, pulverize, add 70% alcohol reflux, concentrate, extractum;

In b, the extractum dissolving entry with a step, filter, filtrate is passed through macroporous adsorptive resins;

C, wash the fat post with water, effluent discards, and when effluent is nearly achromatism and clarity solution, puts solid carbon dioxide, and with 75～90% ethanol elutions, eluent filters, and concentrates and promptly gets total flavones of the present invention;

D, take by weighing the total flavones of c step, add acceptable accessories or complementary composition and be prepared into preparation pharmaceutically commonly used.

Further, above-mentioned antiviral drugs is resisiting influenza virus, parainfluenza virus, adenovirus, its virus of Ke's Sa, respiratory syncytial virus, rhinovirus or herpesvirus medicament.

The present invention also provides the purposes of Rhizoma Iridis Tectori extractive of general flavone in the medicine of preparation analgesia, cough-relieving.

Drug quality of the present invention is stable, controlled, and has determined active ingredient iridin wherein, iristectorin A, iris aglycone, wild flag aglycon, has brought into play the effect of Synergistic by the proportioning of these four compositions.To causing the main virus such as the influenza virus of acute pharyngolaryngitis and upper respiratory tract infection, parainfluenza virus, adenovirus, its virus of Ke's Sa, respiratory syncytial virus, rhinovirus, herpesvirus has inhibitory action significantly, and has significant antiinflammatory, detumescence, antipyretic effect, clinically as the curative of acute pharyngolaryngitis, this drug effect is fast, relief of symptoms is rapid, after taking medicine, the hoarseness person that laryngopharynx swelling and pain causes to recover normal voice in several hours, laryngopharynx swelling and pain obviously alleviates in a few hours or disappears, took medicine continuously 3～7 days, can fully recover, be a kind of active remedy, and cost is low, the yield height provides a kind of new medicinal selection for clinical.

Obviously, according to foregoing of the present invention,,, can also make modification, replacement or the change of other various ways not breaking away under the above-mentioned basic fundamental thought of the present invention prerequisite according to the ordinary skill knowledge and the customary means of this area.

The specific embodiment of form is described in further detail foregoing of the present invention again by the following examples.But this should be interpreted as that the scope of the above-mentioned theme of the present invention only limits to following example.All technology that realizes based on foregoing of the present invention all belong to scope of the present invention.

The specific embodiment

The preparation of embodiment 1 drug extract Rhizoma Iridis Tectori total flavones of the present invention

(1) protogenic medicinal powder is broken into coarse powder and crosses 20 mesh sieves, place in the extraction pot, extract three times with 70% alcohol heating reflux, each 1.5 hours time, each solvent consumption is 4 times (v/w) of medical material amount, and merge extractive liquid, filters, 60 ℃ of decompression recycling ethanols, the syrupy shape material (pale brown color extractum) of proportion 1.2g/ml (50 ℃ of mensuration).Its yield is 49～51% (w/w) of inventory.With above-mentioned extractum 20 times of amount boiling water (w/w) stirring and dissolving with the crude drug amount that is equivalent to feed intake, when treating that solution is cooled to 50 ℃, allow its WLD type macroporous adsorbent resin jar (amount of resin: primary dose 3: 1 by having handled well, w/w), flow speed control is at every 100kg resin per minute 5～10 liters, effluent discards, treat that the reaction of effluent hydrochloric acid magnesium powder shows slightly red and (gets effluent 1ml and put 5ml in vitro, add magnesium powder a little, dripping hydrochloric acid again, it is red that reactant liquor shows), use ordinary water washing resin bed (test of effluent hydrochloric acid magnesium powder is the part of positive reaction and waits until down batch processing) at once, be subdiaphanous clear and bright solution until effluent.Put the ordinary water in the dried resin bed, with 2 times of (w/w) 80% ethanol elutions of amount of resin, flow speed control is at about 5 liters of every 100kg resin per minute, collect eluent, 60 ℃ of decompression and solvent recoveries get pale brown color extractum (proportion 1.2g/ml, measure in the time of 50 ℃, its yield be the medical material amount 29～31%), 60 ℃ of reduced vacuum dryings, pulverize, cross 60 mesh sieves, get pale brown toner powder material (Rhizoma Iridis Tectori extractive of general flavone, content＞55%, water content＜4%, yield are 15～18%.

(2) crude drug in whole coarse powder (20 order) is put in the multipotency extraction pot, extracts three times each 2 hours with 70% alcohol heating reflux.Solvent load is 4 times (v/w) of medical material amount, merges three times extracting solution, filters back 60 ℃ of decompression recycling ethanols, gets pale brown color extractum.This extract stirring is dissolved in 30 times of amounts (w/w) boiling water, treat that solution is cooled to 50～60 ℃, WLD type macroporous adsorptive resins (300 * 60cm) by having handled well while hot, continue with resin in the ordinary water flushing post intact back, discard effluent in the post, when effluent has been clear and bright, put ordinary water in the dried post,, collect eluting ethanol liquid with 90% ethanol elution of 2.5 times of (w/w) amounts of amount of resin, 60 ℃ of decompression and solvent recovery to residues do not have the alcohol flavor, residue carries out spray drying, gets extractive of general flavone yellow powder (cotton-shaped), and content (in iridin) is greater than 55%, water content＜4%, yield are 16～18%.

(3) crude drug in whole coarse powder (20 order) is put in the multipotency extraction pot, extracts four times with 70% alcohol heating reflux, extracts first 3 hours, and the ethanol consumption is 4 times (v/w) of inventory, its excess-three time each extraction 1.5 hours, and solvent load is 3 times (v/w) of inventory.Merge four times extracting solution, the medication film evaporator concentrates, and reclaims ethanol, gets the yellowish-brown paste.With this paste while hot stirring and dissolving in the boiling water of 25 times of amounts, treat that it dissolves complete postcooling to 50～60 ℃, (300 * 60cm), intact back continues to discard effluent in the post with the ordinary water washing resin by the WLD type macroporous adsorptive resins handled well while hot.When treating that effluent has been clear and bright in the post, put ordinary water in the dried post,, collect eluting ethanol liquid, filter with 75% ethanol elution of 3 times of (w/w) amounts of amount of resin.Filtrate is carried out the medicine membrance concentration and is reclaimed ethanol.The extractum that gained has not had the ethanol flavor carries out spray drying, gets the Rhizoma Iridis Tectori total flavones and carries thing (yellow powder), and greater than 55%, water content is less than 4% in iridin for content, and yield is 15～17%.

The preparation of embodiment 2 medicine capsules of the present invention

This product is that the Rhizoma Iridis Tectori extractive of general flavone is directly loaded the hard capsule of making, and every contains the Rhizoma Iridis Tectori total flavones with iridin (C ₂H ₂₂O ₁₁) meter, must not be less than 180mg (every of medicament capsule of the present invention is loaded the extract average weight and is about 0.35g).

Raw material: Rhizoma Iridis Tectori extractive of general flavone 350g is prepared into 1000 medicament capsules of the present invention

(1) dry extractive of general flavone (content＞55%) was pulverized 60 mesh sieves, relative humidity less than 64% condition under, by hand or capsule filling machine load No. 1 capsule, every about 0.35g of control content prepares 1000.

(2) with the Rhizoma Iridis Tectori extractive of general flavone in 60 ℃ of dryings (steam baking house), its water content to be measured＜4% o'clock was pulverized 60 mesh sieves.Accurately take by weighing fine powder 35kg, randomization is an amount of, (general flavone content should be greater than 55% to measure its general flavone content and iridin content by the quality standard method, iridin content should be greater than 18%), if general flavone content surpasses 65%, add an amount of medical starch dilution, make its general flavone content between 55～65%, iridin content is between 18～25%.In the capsule racking room, the control relative humidity is loaded No. 1 capsule with capsule filling machine with medicated powder less than 64%, and every sample-loading amount is 0.35g, prepares 100,000.Thereafter writing paper is pasted in bottling (100/bottle), censorship, vanning warehouse-in.

The preparation of embodiment 3 medicinal tablets of the present invention

Dry extractive of general flavone (content＞55%) was pulverized 60 mesh sieves, add starch, and granulated, granulate, tabletting are promptly.

The quality control of embodiment 4 medicines of the present invention

[discriminating]

(1) gets the about 25mg of this product and place the 50ml volumetric flask,, shake up, as A liquid (0.5ug/ul) with 70% dissolve with ethanol and standardize solution.The accurate A liquid 1ml that draws is in the 50ml volumetric flask, with 70% ethanol dilution and be settled to scale and shake up, as B liquid.B liquid is pressed spectrophotography (2000 editions appendix VA of Chinese Pharmacopoeia), is blank with 70% ethanol, in the interscan of wavelength 200～400nm scope, and the recording light spectrogram, there is absorption maximum at the place at 266 ± 1nm wavelength.

(2) get reference substance iridin 5mg, place the 50ml volumetric flask, add 70% dissolve with ethanol and be diluted to scale, this is product solution in contrast.According to thin layer chromatography (2000 editions appendix VIB of Chinese Pharmacopoeia) test, draw and differentiate (1) item A solution and each 10 μ l of reference substance solution down, parallel in silica gel G F ₂₅₄On the lamellae, be developing solvent with chloroform-methanol-formic acid (10: 1: 0.1), launch that taking-up is dried, (254nm) observes under fluorescent lamp, should show three principal spots, with reference substance chromatograph relevant position on, show identical speckle.

[inspection]

Residue on ignition is got this product 1g, checks (2000 editions appendix IXJ of Chinese Pharmacopoeia) in accordance with the law, and residue on ignition must not cross 2%.

It is an amount of that heavy metal is got this product, after the organic destruction of residue on ignition, checks (2000 editions one appendix IXE second method of Chinese Pharmacopoeia) in accordance with the law, contains heavy metal and must not cross 10/1000000ths.

Arsenic salt is got this product 2.0g, adds water 25ml, checks (2000 editions one appendix IXF second method of Chinese Pharmacopoeia) in accordance with the law, and arsenic content must not cross 2/1000000ths.

Benzene, dimethylbenzene, toluene, styrene, normal heptane, n-decane, divinylbenzene, ethyl styrene

Measure according to gas chromatography (2000 editions appendix VIE of Chinese Pharmacopoeia).

Chromatographic condition and system suitability test chromatographic column adopting HP-INNOWAX post (being PEG 20M), 30m * 0.5mm * 1.0um.Tc＝40℃(5min)→5℃/min→80℃(3min)→20℃/min→200℃(3min)。Detector adopts FID, t=250 ℃, H ₂=35ml/min; Tail blows N ₂=32ml/min; Air=300ml/min.Carrier gas He2.7ml/min, linear velocity S=20cm/s.250 ℃ of the temperature of (constant voltage) injection port.140 ℃ of head space bottle temperature, 150 ℃ of transfer tube temperature, heating equilibration time 10min, head space is split sampling not.

It is an amount of that the preparation precision of reference substance solution takes by weighing normal heptane, benzene, n-decane, toluene, dimethylbenzene, styrene, divinylbenzene, ethyl styrene, with dimethyl sulfoxine dissolving and standardize solution (2.5mg/ml) in the 10ml volumetric flask.Accurate again this solution of absorption 10 μ l are settled to scale with dimethyl sulfoxine in the 10ml volumetric flask, shake up (2.5 μ g/ml).Draw this and be dissolved in 1ml and be encapsulated in the head space bottle, reference substance solution.

It is an amount of that this product is got in the preparation of need testing solution, and accurate the title decides.Be encapsulated in the head space bottle with the 1ml dimethyl sulfoxine, for measuring (0.5g/ml).

Algoscopy is drawn reference substance solution and each 500 μ l of need testing solution respectively, and in the inject gas chromatograph, the record chromatogram is made blank with dimethyl sulfoxine simultaneously, calculates promptly.

This product contains toluene, dimethylbenzene, styrene, normal heptane, n-decane, divinylbenzene, ethyl styrene must not cross 2/100000ths, and benzene must not cross 2/1000000ths.

[loss on drying] gets this product, is dried to constant weight at 105 ℃, reduces weight and must not surpass 6% (2000 editions appendix IXG of Chinese Pharmacopoeia).

[assay]

Total flavones

The accurate title of the preparation of reference substance solution fixes on 105 ℃ of iridin reference substance 15mg that are dried to constant weight, places the 50ml volumetric flask, adds 70% dissolve with ethanol and is settled to scale, shakes up, promptly.Every 1ml contains iridin 0.3mg.

The preparation precision of need testing solution takes by weighing 105 ℃ of about 25mg of this product that are dried to constant weight, places the 50ml volumetric flask, adds 70% dissolve with ethanol and is settled to scale, shakes up, promptly.Every 1ml contains this product 0.5mg.

Accurate reference substance solution and the need testing solution 1ml of drawing of algoscopy places the 50ml volumetric flask respectively, with 70% ethanol dilution and be settled to scale, shakes up.Pressing spectrophotography (2000 editions appendix VB of Chinese Pharmacopoeia), at 266nm wavelength place, is blank with 70% ethanol, measures the trap of reference substance and need testing solution respectively, calculates, promptly.

This product is calculated by dry product and is contained total flavones with iridin (C ₂₂H ₂₂O ₁₁) calculate, must not be less than 55%.

Iridin

Measure according to high performance liquid chromatography (2000 editions appendix VID of Chinese Pharmacopoeia).

Chromatographic condition and system suitability test octadecylsilane (C ₁₈) bonded silica gel is filler, methanol-0.5% acetic acid (30: 70) is mobile phase; The detection wavelength is 266nm; Column temperature is a room temperature, and theoretical cam curve is pressed reference substance iridin peak and calculated, and should be not less than 7000.

The preparation precision of reference substance solution takes by weighing the iridin reference substance 5mg that is dried to constant weight, places in the 50ml measuring bottle, adds the about 40ml of 70% methanol, complete with supersonic oscillations 5～10min to dissolving, continue and add 70% methanol, shake up, promptly get (containing iridin 0.1mg among every ml) to scale.

The about 20mg of Rhizoma Iridis Tectori total flavones that is dried to constant weight is got in the preparation of need testing solution, and accurate the title decides.Place the 50ml measuring bottle, add 70% methanol solution and be mixed with solution, shake up promptly according to method under " reference substance solution preparation " item.

Accurate respectively need testing solution and each the 10 μ l of reference substance solution of drawing of algoscopy inject hplc determination, promptly.

This product is calculated by dry product and is contained iridin (C ₂₂H ₂₂O ₁₁), must not be less than 18%.

Below prove drug effect of the present invention by pharmacodynamics test.

Test example 1 iris aglycone extracorporeal antivirus effect laboratory report

One, experiment material

(1) is subjected to reagent

Iris aglycone (iris aglycone) is made the 25mg/ml sterile solution, is provided by the Traditional Chinese Medicine Research Institute, Sichuan Province.

(2). positive control drug

Virazole (ribavirin glycoside injection liquid), the accurate word XF1990722 of traditional Chinese medicines, lot number 000402,100mg/ml, Hubei Co., Ltd of suitable medicine group produces.

Injection Andrographolide, river are defended the accurate word (1981) of medicine No. 006085, lot number 981201, and pharmacy three factories in Chengdu produce 40mg/ml.

(3). cell strain

HeP-2 is provided by Sichuan Province health and epidemic prevention station, and my chamber is frozen, recover, go down to posterity, cell growth medium is 10% calf serum Eagles liquid.

Z-HL ₁₆The C cell, biological engineering Applied Research Laboratory cell engineering research department provides by Shantou, and go down to posterity in my chamber, and cell growth medium is 10% calf serum Eagles liquid.

(4). Strain

Adenovirus (AdV _3,7,) respiratory syncytial virus (RSV), change of coxsackie b virus (CoxBV), herpes simplex virus (HSV ₁) and influenza virus ( _{First 1, first 3}) provide by Shoudu Inst. of Pediatrics and Institute of Viruses, Chinese Academy of Medical Sciences respectively, go down to posterity in my chamber, experiment before measurement TCID ₅₀

Two, experimental technique

(1) cytotoxicity experiment

Get iris aglycone solution (25mg/ml) and make doubling dilution with Hanks liquid, make 4 concentration liquid cytotoxicity experiments (being 25mg/ml, 12.5mg/ml, 6.25mg/ml, 3.125mg/ml), begin to do 4 concentration liquid antiviral experiments with avirulence cell drug concentration then.Getting positive control drug virazole, injection Andrographolide synchronously is diluted to 4 concentration liquids (25mg/ml, 12.5mg/ml, 6.25mg/ml, 3.125mg/ml) with Hanks liquid and gets the various variable concentrations inoculation of above 3 medicinal liquids Hep-2, Z-HL respectively ₁₆Each 2 pipe of C cell fill up Eagles liquid and keep liquid (establishing cell contrast and blank synchronously), put 37 ℃ of incubators, day by day the observation of cell toxic reaction.

(2) antiviral experiment

Adopt direct deactivation method, determine before the experiment that each viroid causes cell half pathological changes amount (TCID ₅₀/ 0.1ml) and various kinds of drug pair cell non-toxic reaction boundary line, get 100TCID ₅₀And AdV _3,7,, RSV, CoxBV, HSV ₁And influenza _{First 1, First 3}After virus directly acts on 1 hour with the medicinal liquid mixed in equal amounts of above-mentioned 3 kinds of all kinds of variable concentrations of medicine respectively, get each viroid respectively and medicament mixed liquid is inoculated Hep-2 and Z-HL respectively ₁₆The C cell, after waiting to adsorb 45 minutes, fill up cell maintenance medium (2% calf serum Eagles), establish cell contrast, virus control and Hanks liquid synchronously and make blank, putting 37 ℃ of incubators cultivates, (RSV inactivation of virus experiment tube moves after 24 hours and puts 33 ℃ of rotary incubators), observation of cell pathological changes day by day.When the 3+ pathological changes appears in virus control, cell contrast well-grown, form is normal, but termination test.Whether each viroid is suppressed by medicine, and pathological changes whether occurring with cell is index, when 2+ pathological changes (〉=50% cytopathy) appears in cell, can judge that this medicinal liquid is to viral unrestraint effect.

(3) experimental result

The cytotoxicity experiment result

Iris aglycone solution, ribavirin injection, injection Andrographolide 3 kinds of medicine 25mg/ml, 12.5mg/ml, 6.25mg/ml, 3.125mg/ml4 kind concentration liquid are to Hep-2, Z-HL ₁₆The C cell does not all have any toxic reaction.Blank, cell contrast equal avirulence, the cell well-grown, and form is normal.

The antiviral experimental result

Iris aglycone solution 25mg/ml-12.5mg/ml medicinal liquid is to AdV _3,7, CoxBV and influenza _{First 3}Virus, and the 25mg/ml-6.25mg/ml medicinal liquid to RSV ₁Virus all can suppress virus fully and cause pathological changes effect in the cell.

Ribavirin injection 25mg/ml-12.5mg/ml medicinal liquid is to influenza _{First 1, first 3}, AdV ₃Pathological changes effect in the cell of inhibition is all arranged, and the 25mg/ml-3.125mg/ml medicinal liquid is to RSV, and the 25mg/ml-12.5mg/ml medicinal liquid is to CoxBV, AdV ₇And HSV ₁Pathological changes effect in the cell of inhibition is all arranged, and the 6.25mg/ml medicinal liquid is to AdV ₇And CoxBV all has the pathological changes effect in the cell that partly suppresses.

Potasium dehydroandrographolisuccinate succinate injection 25mg/ml-12.5mg/ml medicinal liquid is to AdV _3,7Pathological changes effect in the cell of inhibition is all arranged, and the 25mg/ml medicinal liquid all has pathological changes effect in the cell of inhibition to CoxBV and RSV, and the 12.5mg/ml medicinal liquid has the pathological changes effect in the cell that partly suppresses to CoxBV and RSV.Blank all can not suppress AdV ₃Deng pathological changes effect in the virocyte.Blank all can not suppress AdV ₃Cause pathological changes effect in the cell etc. virus.

The experiment of iris aglycone solution extracorporeal antivirus effect is carried out with blank under the identical situation of drug level synchronously with positive control drug virazole, Andrographolide, and all adopt direct deactivation method, its results suggest: iris aglycone solution is to AdV _3,7Type, influenza _{First 3}Type, CoxBV and RSV all have inhibitory action in various degree, and the lowest concentration of drug that RSV is suppressed is 6.25mg/ml.Ribavirin injection is to AdV ₃7 kinds of viruses such as type all have inhibitory action, can reach 0.3125mg/ml to most viral inhibiting lowest concentration of drug.The injection Andrographolide is to AdV _3,7, CoxBV and RSV have inhibitory action in various degree, to AdV _3,7The inhibiting lowest concentration of drug of type can reach 12.5mg/ml.

This results suggest: ribavirin injection all is better than iris aglycone and Andrographolide to the inhibitory action of 7 kinds of viruses such as AdV.Iris aglycone solution is to influenza _{First 3}The inhibitory action of type, CoxBV and RSV all is better than Andrographolide.

The experiment of test example 2 iridin extracorporeal antivirus effects

1, experiment material

(1) medicine: iridin (iridin)

Provided by the Traditional Chinese Medicine Research Institute, Sichuan Province by reagent iridin (white powdery).Be made into 100mg/ml stock solution with Hanks liquid before the experiment.(add the inferior maple of cosolvent dimethyl and help dissolving) degerming.PH was about 7, and stock solution was made the proportional diluted of 7 variable concentrations since 1: 10, and its medicament contg is 10mg/ml, 5mg/ml, 2.5mg/ml, 1.25mg/ml, 0.62mg/ml, 0.31mg/ml, 0.15mg/ml, and is standby.

Positive control drug virazole (ribavirin glycoside injection liquid 100mg/ml), the accurate word (1991) of medicine is defended No. 000042 in Hubei Province, lot number 980605, the Yichang three gorges pharmaceutical factory produces, I provide in the institute pharmacy, be made into 10mg/ml, 5mg/ml, 2.5mg/ml, 1.25mg/ml, 0.62mg/ml, 0.31mg/ml, 0.15mg/ml with Hanks liquid before the experiment, standby.Positive control drug potasium dehydroandrographolisuccinate succinate injection (40mg/ props up dried frozen aquatic products), river are defended the accurate word (1981) of medicine No. 000085, lot number 981201, pharmacy three factories in Chengdu produce, I provide in the institute pharmacy, am mixed with and iridin, 7 medicinal liquids that concentration is identical with Hanks liquid before the experiment, and am standby.

Blank replaces medicinal liquid and xy, virazole, the experiment of Andrographolide antiviral to carry out synchronously with Hanks liquid.

(2) cell strain

Hela, HeP-2, MDCK (Testis et Pentis Canis passage cell) grind institute, Sichuan Province health and epidemic prevention station and Beijing Shoudu Inst. of Pediatrics by the Chengdu life respectively and provide, and my chamber recovery is gone down to posterity, and growth-promoting media is 10% calf serum Eagles liquid.

(3) Strain

Adenovirus (ADV) _3,7,11Type, respiratory syncytial virus (RSV), influenza virus _{First 1}Type _{First 3}Type (FluA ₁A ₃), herpes simplex virus (HSV) ₁Type, ₂Type, change of coxsackie b virus (CoxBV) is mixed strain, is provided by Beijing Shoudu Inst. of Pediatrics, Institute of Viruses, Chinese Academy of Medical Sciences and Sichuan Province health and epidemic prevention station respectively, and go down to posterity in my chamber, experiment before measurement TCID ₅₀

2, experimental technique

(1) cytotoxicity experiment

Get iridin, virazole, 7 kinds of variable concentrations medicinal liquids of Andrographolide medicinal liquid and inoculate Hela, HeP-2, mdck cell respectively, after absorption, fill up 2% calf serum Eagles liquid and keep liquid, put 37 ℃ of incubators, day by day the observation of cell toxic reaction.Establish the cell contrast synchronously.

(2) extracorporeal antivirus effect experiment

Get the medicinal liquid of iridin, virazole, 7 kinds of variable concentrations of Andrographolide medicinal liquid and blank Hanks liquid respectively with 100TCID ₅₀And AdV _3,7,11Type, RSV, HSV ₁, HSV ₂, FluA ₁, A ₃And the CoxBV mixed in equal amounts, under room temperature, directly act on 1 hour, after (establishing each viroid synchronously, the cell contrast), get various kinds of drug, Hanks and viral mixed liquor, inoculation HeP-2 cell (anti-FluA ₁A ₃Inoculation mdck cell, anti-RSV inoculation Hela cell), after waiting to adsorb 45 minutes, fill up 2% calf serum and keep liquid, all put 37 ℃ of incubators, anti-RSV tests to move next day to be put outside 35 ℃ of rotary incubators, and other antiviral cells Guan Jun continues in 37 ℃ of environment and cultivates, day by day observation of cell pathological changes, contrast well-grown when 3+ pathological changes (〉=75% cytopathy) cell appears in virus control, form is normal, but termination test.Whether each viroid is suppressed by medicine, and pathological changes whether occurring with cell is index, and 2+ pathological changes (〉=50% cytopathy,＜75% cytopathy=can judge that this medicinal liquid is invalid to virus) appears in all cells.

3, experimental result

(1) cell toxicity test

Be subjected to reagent iridin 10mg/ml medicinal liquid to Hep-2, mdck cell can cause partly that the cell circle contracts, shrinkage, come off outside, other 6 kinds of concentration liquids are to Hela cell non-toxic reaction.

Contrast medicine virazole, 7 kinds of variable concentrations medicinal liquids of Andrographolide all do not have any toxic reaction to Hela, Hep-2, mdck cell.

(2) antiviral experimental result

Be subjected to reagent iridin 5mg/ml-10mg/ml medicinal liquid, contrast medicine virazole 0.31mg/ml-10mg/ml and Andrographolide 2.5-10mg/ml medicinal liquid are to AdV ₃9 kinds of viruses such as type all have the interior pathological changes effect of inhibition cell in various degree, and blank all can not suppress AdV ₃Pathological changes in 9 kinds of virocytes such as type.

Conclusion

Iridin 10mg/ml medicinal liquid is to AdV ₃Type, HSV ₁9 kinds of viruses such as type all have inhibitory action, and the 5mg/ml medicinal liquid is to AdV ₇Type, RSV and HSV ₂9 kinds of viruses such as type all have inhibitory action.Other concentration liquid is to AdV _3,7The equal unrestraint effect of type.

Virazole 2.5mg/ml-10mg/ml medicinal liquid is to AdV ₃9 kinds of viruses such as type all have inhibitory action, and the 1.25mg/ml medicinal liquid is to AdV ₁₁Type, RSV, AdV ₃Type, CoxBV and FluvA ₁6 kinds of viruses all have inhibitory action.Other concentration liquid AdV _3,7The equal unrestraint effect of type.0.62mg/ml with the 0.31mg/ml medicinal liquid RSV is had inhibitory action, other concentration liquids are to Adu ₃Unrestraint effects such as type.

Andrographolide 10mg/ml medicinal liquid is to AdV ₃9 kinds of viruses such as type all have inhibitory action, and the 5mg/ml medicinal liquid is to AdV ₃Type, AdV ₁₁Type, RSV, HSV ₂Type, CoxBV have inhibitory action.2.5mg/ml medicinal liquid is to FluVA ₁, A ₃Inhibitory action is all arranged.Other concentration liquid is to AdV ₃The equal unrestraint effect of type.

Above results suggest is all had in various degree to AdV by reagent iridin, contrast medicine virazole, Andrographolide ₃9 kinds of viruses such as type have inhibitory action, and virazole is to AdV ₃The antivirus action of 9 kinds of viruses such as type more obviously is better than iridin and Andrographolide.

Test example 3 iristectorin A medicine extracorporeal antivirus effect laboratory reports

1. experiment material

(1). medicine: provided by the Traditional Chinese Medicine Research Institute, Sichuan Province by reagent iristectorin A (iristectorin A, white powdery).Be made into 100mg/ml stock solution with Hanks liquid before the experiment.(add dimethyl sulfoxide and help dissolving) sterilization degerming.PH is about 7, with stock solution from 1: 10-1: 160 make doubling dilution, and 1: 10 medicinal liquid is 10mg/ml.Positive control drug, virazole (ribavirin glycoside injection liquid 100mg/ml), the accurate word (1991) of medicine is defended No. 000042 in Hubei Province, lot number 980605, the Yichang three gorges pharmaceutical factory produces, I provide in the institute pharmacy, the experiment before be made into 10mg/ml stock solution with Hanks liquid, with stock solution from 1: 10-1: 160 make doubling dilution, and 1: 10 medicinal liquid is 1mg/ml.Positive control drug, potasium dehydroandrographolisuccinate succinate injection (40mg props up dried frozen aquatic products), river are defended the accurate word (1981) of medicine No. 006085, lot number 981201, pharmacy three factories in Chengdu produce, I provide in the institute pharmacy, and do 1 with Hanks liquid before the experiment: 10-1: 160 make doubling dilution, and 1: 10 medicinal liquid is 4mg/ml.Blank replaces medicinal liquid with Hanks liquid, carries out with being subjected to reagent, the contrast of positive Chinese medicine and western medicine synchronously.

(2). cell strain: Hela, HeP-2, MDCK (Testis et Pentis Canis passage cell) grind institute, Sichuan Province health and epidemic prevention station and Beijing Shoudu Inst. of Pediatrics by the Chengdu life respectively and provide, and I recover the chamber, go down to posterity, growth-promoting media is 10% calf serum Eagles liquid.

(3). Strain: adenovirus (AdV) _3,7,11Type, respiratory syncytial virus (HSV), influenza virus (Flu.V) first ₁The type first ₃Type, herpes simplex virus (HSV) 1 type, 2 types, change of coxsackie b virus (CoxBV) is mixed strain, is provided by Beijing Shoudu Inst. of Pediatrics, China prevention medical courses in general institute institute of viruses, Beijing and Sichuan Province health and epidemic prevention station respectively, and go down to posterity in my chamber, experiment before measurement TCID ₅₀

2. experimental technique

(1). cytotoxicity experiment

Get 1: 10-1: the iristectorin A of 160 variable concentrations, virazole, Andrographolide medicinal liquid, respectively inoculate Hela, HeP-2, mdck cell, fill up 2% calf serum Eagles liquid and keep liquid, put 37 ℃ of incubators, observe the various types of cells toxic reaction day by day.Establish the various types of cells contrast synchronously.

(2) antiviral experiment

Get 1: 10-1: the direct respectively and AdV of the medicinal liquid of 80 4 iristectorin As, virazole, Andrographolide medicinal liquid, various kinds of drug variable concentrations with concentration _3,7,11Type, RSV, HSV ₁, HSV ₂The Flu first ₁The type first ₃Mixing such as type and CoxB papova directly act on 1 hour under room temperature, (establish each viroid synchronously, various types of cells contrast and blank (replacing medicine with hanks liquid) are got various kinds of drug variable concentrations medicinal liquid and all kinds of different virus mixed liquor, remove the Flu first ₁The type first ₃Type inoculation mdck cell, RSV inoculation Hela extracellular, other viral mixed liquor are all inoculated the HeP-2 cell, fill up 2% calf serum and keep liquid, putting 37 ℃ of incubators cultivates, removing anti-RSV test cultivates after 24 hours for 37 ℃, move and put outer other antivirus tests of 35 ℃ of rotary incubators (12 change/hour) and all continue to put 37 ℃ of cultivations, observation of cell pathological changes day by day is when 3+ (〉=75% cytopathy) cell contrast well-grown appears in virus control, form is normal, but termination test.Whether each viroid is suppressed, and pathological changes whether occurring with cell is index, can judge that when 2+ pathological changes (〉=75% cytopathy) appears in various types of cells this medicinal liquid is invalid to virus.

3. experimental result

(1). cell toxicity test result

Outside 1: 10 medicinal liquid of iristectorin A medicine can cause partly to Hep-2, mdck cell that the cell circle contracts, shrinkage comes off, 1: 10-1: 160 medicinal liquids did not all have any toxic reaction to the Hela cell.

Contrast medicine virazole, Andrographolide 1: 10-1: 160 medicinal liquids all do not have any toxic reaction to Hela, Hep-2, mdck cell.

(2). the antiviral experimental result

Get 1: 10 medicinal liquid of iristectorin A to AdV ₃Type, ₁₁Type, HSV ₁Type, CoxBV and influenza _{First 1}Type, _{First 3}Type is no matter all can suppress pathological changes in the cell to which kind of cell, and medicinal liquid was to AdV in 1: 20 ₇Type, RSV and HSV all can suppress cytopathy; 1: 10 medicinal liquid of virazole is to AdV ₃Type, ₁₁Type, CoxBV and influenza _{First 1}Type, _{First 3}Type all has pathological changes effect in the cell of inhibition.To HSV ₁Type has the pathological changes effect in the cell that partly suppresses, and medicinal liquid was to HSv in 1: 20 ₂Type has partly inhibitory action, and medicinal liquid had pathological changes effect in the cell of inhibition to RSV in 1: 40.1: 10 medicinal liquid of Andrographolide is to AdV ₁₁Type, HSV ₂Type, 1: 20 medicinal liquid are to AdV ₃Type, RSV ₁And influenza _{First 1}Type, _{First 3}Type all has pathological changes effect in the cell of inhibition.To AdV ₇Type, HSV ₁The equal unrestraint effect of type and CoxBV.

4. conclusion

Iristectorin A 1: 10 medicinal liquid (10mg/ml) is to AdV ₃₁₁Type, HSV ₁Type CoxBV, influenza _{First 1}Type, _{First 3}Type all has the pathological changes effect in the cell that suppresses, 1: 20 medicinal liquid (5mg/ml) to AdV ₇Type, RSV, HSV ₂Inhibitory action is all arranged.

Western medicine contrast virazole 1: 10 medicinal liquid (1mg/ml) is to AdV ₃₁₁Type, HSV ₁Type CoxBV, influenza _{First 1}Type, _{First 3}Type all has inhibitory action, to HSV ₁Type has partly inhibitory action.1: 20 medicinal liquid (0.5mg/ml) to HSV ₂Partly inhibitory action is arranged.1: 40 medicinal liquid (0.25mg/ml) RSV is had inhibitory action.1: 10-1: 80 medicinal liquid AdV ₇Type unrestraint effect.

Chinese medicine contrast Andrographolide 1: 10 medicinal liquid (4mg/ml) is to AdV ₃₁₁Type, RSV, HSV ₂Type and influenza _{First 1}Type, _{First 3}Type has inhibitory action.1: 20 medicinal liquid (2mg/ml) to AdV ₃Type, RSV, influenza _{First 1}Type, _{First 3}Type all has inhibitory action, 1: 10-1: 80 medicinal liquid AdV ₇Type, HSV ₁Type, the equal unrestraint effect of CoxBV.

Blank all can not suppress AdV _3,7,11Type, RSV, HSV ₁, HSV ₂Type, influenza _{First 1}Type, _{First 3}Type virus.

The iristectorin A medicine in the vitro tissue cell to AdV _3,7,11Type, RSV, HSV _1,2Type CoxBV, influenza _{First 1}Type, _{First 3}Type all has inhibitory action in various degree.Wherein to AdV ₇, HSV ₂The type inhibitory action all is better than virazole and Andrographolide.

Virazole all is better than iristectorin A and Andrographolide medicine to the inhibitory action of RSV.

Andrographolide is to influenza _{First 1}Type, _{First 3}The inhibitory action of type all is better than iristectorin A medicine and virazole.

Below be pharmacodynamics test be subjected to reagent thing, instrument medicine positive drug:

1, is subjected to the reagent thing

Medicament capsule of the present invention is a capsule, by embodiment 3 preparations, the amount of setting medicated powder 0.35g in the capsule.Clinical application is drafted day clothes 3 times, 1 time 3.Acute pharyngolaryngitis 1 week of the course of treatment, 2 months courses of treatment of chronic pharyngolaryngitis.Become for each person in 60 kg body weight, then day kg body weight dosage is 0.05g/kg.The test medication is that pilot scale makes and is equipped with encapsulated medicated powder, is yellow powder.Provide by Traditional Chinese Medicine Research Institute, Sichuan Province Chemistry for Chinese Traditional Medicine Xu Xuemin researcher of research department etc.Be made into desired concn with 0.5% tragcanth solution before the test, g/ml represents with medicated powder weight, with medicated powder weight g/kg b.w. (body weight) expression dosage.The live part content of each batch medicated powder is slightly different, and each batch can not surpass ± 3% (promptly 60 ± 3%) on average about 60%, with this as quality control standard, with the stability of maintenance drug effect.This batch of test specimen live part content is 59.55%.

2, instrument medicine, positive control drug, important reagent, culture medium etc.

Positive control drug:

Ribavirin injection: (100mg/ml), the accurate word (1983) of medicine is defended No. 000229 in Hubei Province, and Tianmen, Hubei pharmaceutical factory produces, lot number: 990610.Make 1: 10～1: 80 proportional diluted during test.

Andrographolide: lyophilized injectable powder, 40mg/ props up, and Tong De pharmaceutcal corporation, Ltd in Chengdu produces, lot number: 991101.Be diluted to 100mg/ml before the test, remake 1: 10～1: 80 proportional diluted.

Aspirin: the packing of Chongqing company of China Drug Co., lot number: 990507

Watermelon crystal: Guilin three gold medal Pharmaceutical group companies produce, lot number: 9904248

Hydrocortisone: South-west Pharmaceutical Factory No.3 produces, lot number: 991205

Larynx disease curing capsule: Baiyunshan Pharmaceutical General Factory pharmaceutical factory in spring produces, lot number: 980201.Guangzhou Chenliji Pharmaceutical Factory produces, lot number: 9907314

Morphine hydrochloride: Shenyang No. 1 Pharmaceutical Factory production, lot number: 990802

JINSANGZIHOUBAO: Guangxi Golden Throat Pharmaceutical Factory is produced, lot number: 991001

Codeine phosphate: Qinghai Pharmaceutic Plant of China Drug Co. produces.

JIZHI TANGJIANG: Tai Ji group Fuling Pharmaceutical Factory is produced, lot number: 001016095

Other reagent medical instruments: 0.6% HAC physiological salt liquid; 0.5% azovan blue physiological salt liquid; 0.9% normal saline, PH7.0～7.2; Dimethylbenzene (AR), the autoclaving cotton balls; Caseinhydrolysate fluid medium (MH culture medium) contains 3% serum MH culture medium

3. laboratory animal, cell, virus

Mice: Kunming kind outbreeding system (closed colony is more than 30 generations), 1 grade of qualified animal, the quality certification number is No. the 30th, the real moving pipe matter (99) in river.Body weight is generally selected between 18～22g.Male and female half and half.Number of animals is decided according to experiment content, is provided by Sichuan Province's antibiotics industrial research institute animal center.

Rat: SD kind outbreeding system (closed colony is more than 30 generations), 1 grade of qualified animal, the quality certification number: No. the 32nd, the real moving pipe matter (99) in river.Body weight is generally selected between 180～220g.Male and female half and half.Number of animals is decided (consulting the experimental technique part) according to experiment content, provided by Sichuan Province's antibiotics industrial research institute animal center.

Cell strain: the Hep-2 cell, frozen by clinical laboratory of People's Hospital, Sichuan Prov. Viral Laboratory-85 ℃ low temperature, recovery is gone down to posterity before the experiment, and growth-promoting media is 10% calf serum Eagles liquid, treats that cell is standby after test tube forms monolayer.

Virus: COxsackie CoxB3 type, adenovirus AdV3 type, Shoudu Inst. of Pediatrics provides by Beijing, and recovery is gone down to posterity in clinical laboratory of People's Hospital, Sichuan Prov. Viral Laboratory, goes down to posterity respectively before the test to measure ICID ₅₀/ 0.1ml.

The antivirus test research of test example 4 medicines of the present invention

1. cell toxicity test

Get medicine of the present invention, virazole, 1: 10～1: 80 medicinal liquid of three kinds of medicines of Andrographolide and (be equivalent to 10,5,2.5,1.25mg/ml), be inoculated in Hep-2 cell pipe respectively, add 2% calf serum Eagles liquid and keep, put 37 ℃ of incubators, establish the cell contrast synchronously, observation of cell reaction day by day.The result can cause a little shrinkage except that 1: 10 pair of Hep-2 cell of medicine medicinal liquid of the present invention, and medicinal liquid did not all have any toxic reaction to the Hep-2 cell in 1: 20～1: 80, and virazole, Andrographolide all do not have any toxic reaction to the Hep-2 cell.The results are shown in Table 1.

Table 1 medicine of the present invention and contrast medicine pair cell toxicity test result

Annotate :-cell does not have any toxic reaction

A little shrinkage appears in ± expression

The result shows: medicine 5,2.5 of the present invention, and 1.25mg/ml is to Hep-2 cell avirulence, and 10mg/ml has mild toxicity to Hep-2 cell 3.Andrographolide, virazole 10,5,2.5,1.25mg/ml is to the equal avirulence of Hep-2 cell.

2. external direct antivirus test

Get medicine of the present invention, virazole, four concentration liquids of three kinds of medicines of Andrographolide (1: 10,1: 20,1: 40,1: 80), respectively with 100TCID ₅₀The CoxB3 of/0.1ml, AdV3 virus mixed in equal amounts is put room temperature and is directly acted on 1 hour.(set up separately simultaneously virus control, cell contrast) gets every kind of medicine variable concentrations liquid and CoxB3, AdV3 equivalent mixed liquor 0.2ml, inoculate 2 Hep-2 cell pipes (establishing virus control inoculation 0.1ml synchronously) respectively, liquid to be mixed, virus absorption onto cell are after 45 minutes, fill up in each pipe and to keep liquid, put 37 ℃ of incubators, day by day observation of cell pathological changes situation treats to end when the 3+ pathological changes appears in virus control test.All cells 2+ occurs or the above pathological changes of 2+＞50% promptings are subjected to reagent can not suppress virus; Pathological changes does not appear in cell, and the prompting medicine has inhibitory action to virus; The following pathological changes of 1+ appears in cell, and the prompting medicine has part to suppress virus function.Result of the test shows: under similarity condition, blank cell well-grown does not have pathological changes, and serious pathological changes (3+) appears in the virus control cell, medicine of the present invention 1: 10,1: 20,1: 40 (promptly 10,5,2.5mg/ml) medicinal liquid can suppress the CoxB3 cytopathogenic effect fully; And medicine of the present invention 1: 10 (being 10mg/ml) suppresses the AdV3 cytopathogenic effect fully.Western medicine contrast medicine virazole just 1: 10 medicinal liquid (being 10mg/ml) can suppress the pathological changes that CoxB3 causes, and to the AdV3 cytopathogenic effect, 1: 20,1: 40,1: 80 (promptly 5,2.5,1.25mg/ml) can not suppress cytopathy.(promptly 10,5,2.5mg/ml) medicinal liquid can suppress the CoxB3 cytopathogenic effect, and each experimental concentration all can not suppress the AdV3 cytopathogenic effect for Chinese medicine positive drug Andrographolide 1: 10,1: 20,1: 40.Above result shows: medication medication of the present invention has antivirus action in vitro tests, and Coxsackie virus (CoxB3) is better than the antivirus action of virazole, and is consistent with the effect of Andrographolide; The effect of adenovirus (AdV3) is better than the antivirus action of Andrographolide, consistent with the effect of virazole.More than the results are shown in Table 2.

The antivirus action of table 2 medicine of the present invention

Annotate :-expression cellular morphology is normal, no pathological changes

(-) expression medicine suppresses cytopathy fully

(++)～(+++) expression cytopathy degree

3. cell toxicity test

Use Hank liquid before the experiment, medicine of the present invention (SG-2), Western medicine contrast medicine virazole, Chinese medicine contrast medicine Andrographolide three medicines were made proportional diluted with 1: 10～1: 160, Cmax is 10mg/ml, is 5,2.5,1.25 successively, variable concentrations such as 0.625mg/ml.Get the variable concentrations medicinal liquid of above three medicines respectively, be inoculated in the HeLa cell, mdck cell is established the cell contrast that does not add medicine simultaneously respectively, puts 37 ℃ of incubators, day by day the reaction of observation of cell poison.The result shows: medicine of the present invention (SG-2), virazole, Andrographolide and the blank (Hanks liquid) of 1: 10～1: 60 variable concentrations of medicine of the present invention are to HeLa, and mdck cell does not all have any toxic reaction.See Table 3.

Table 3 medicine of the present invention and contrast medicine pair cell toxicity test result

Annotate :-expression 2 test tube (parallel multiple pipe) all do not have any toxicity

4. antivirus test

Get 100TCID ₅₀The respiratory syncytial virus of/0.1ml (RSV), influenza virus A 1 type, first 3 types respectively with medicine of the present invention (SG-2), virazole, 1: 10～1: 80 medicinal liquid mixed in equal amounts of Andrographolide, after the room temperature effect 1 hour, get above-mentioned each viroid and medicament mixed direct inoculation HeLa and mdck cell respectively, treat that cell absorption after 1 hour, adds 10% calf serum growth-promoting media respectively, establish virus, cell contrast and blank synchronously, put 37 ℃ of incubators, day by day observation of cell pathological changes situation.

Experimental result shows: medicine of the present invention 1: 10; 1: 20 medicinal liquid; be 10mg/ml; 5mg/ml is to respiratory syncytial virus (RSV); influenza virus A 1 type, and 1: 10, promptly 10mg/ml all can suppress cell virus for influenza virus A 3 types; show that this concentration has antivirus action, pair cell has protection, and it avoids the effect of virus infraction.1: 40 medicinal liquid of virazole, promptly 1.25mg/ml is to RSV virus, and 1: 10mg/ml has the inhibition cytopathic effect to influenza virus A 1, first 3 types.1: 20 medicinal liquid of Andrographolide is promptly: 5mg/ml all has its virus of inhibition to RSV, influenza first 1, first 3 type viruses, and the protection cell is avoided the effect of virus damage.The results are shown in Table 4.

The antivirus action of table 4 medicine of the present invention

Annotate: "-" expression 2 solencytes do not have any pathological changes

" ++--+++" expression cytopathy degree

Above-mentioned antiviral experimental results show that, medicine of the present invention has inhibitory action significantly to the main virus that causes acute pharyngolaryngitis and acute upper respiratory tract infection such as influenza virus, adenovirus, its virus of Ke's Sa, respiratory syncytial virus, its effect is suitable with the sick virazole of Western medicine, be better than commercially available medicine Andrographolide (andrographolide succinic acid half-ester monopotassium salt is treatment by Chinese herbs acute upper respiratory tract infection drug of first choice).

The experimental study of test example 5 medicine antiinflammations of the present invention

1, medicine Dichlorodiphenyl Acetate induced mice of the present invention abdominal cavity capillary permeability increases the influence test of (inflammation)

Get 60 of body weight 20～25g mices, be divided into 6 groups at random, gavage medicine 1.0 of the present invention respectively by body weight, 0.5,0.25g/kg, Western medicine contrast medicine group aspirin 0.1g/kg, Chinese medicine contrast medicine group watermelon crystal sheet 1.2g/kg, the blank group gavages isopyknic 1% tragakanta solution.More than each group all after administration 1 hour, each caudal vein injection azovan blue normal saline 0.1ml/10gbw, lumbar injection 0.6% glacial acetic acid (HAC) 0.2ml/ only immediately, after 20 minutes mice being pulled cervical vertebra puts to death, cut off skin of abdomen muscle, divide several to wash the abdominal cavity with the 6ml normal saline, suction pipe sucking-off cleaning mixture, merge and add normal saline to 10ml, the centrifugal 15min of 3000rpm, get supernatant in UV-730 biochemistry analyzer 590nm place's colorimetric determination, represent permeability, the result is carried out statistical with the OD value.Result such as table 5.

Table 5 medicine medicated powder of the present invention is to the influence of mouse peritoneal permeability (X ± SD)

Annotate: ^*: P＜0.01, ^{* *}: P＜0.001 (comparing) with matched group

Experimental result shows: medicine 1.0 of the present invention, 0.5,0.25g/kg lumbar injection HAC induced mice abdominal cavity capillary permeability is increased the inhibitory action that highly significant is arranged, the extremely significantly significant difference (P＜0.01) of having compared with matched group shows that medicine medicated powder of the present invention has the effect of this kind of inhibition inflammation of highly significant.Its effect is than strong with the dosage watermelon crystal.

2, medicine xylol of the present invention causes the influence of mice auricle swelling

Get 60 of body weight 18～22g mices, male and female half and half, be divided into 6 groups at random by body weight, 3 administration groups are irritated stomach medicine 1.0 of the present invention respectively, 0.5,0.25g/kg, Western medicine positive control drug group sc hydrocortisone 25mg/kg, Chinese medicine contrast medicine group ig larynx disease spirit 0.66g/kg, blank group ig equal-volume 1% tragakanta solution.Each group is respectively in the ig administration after 30 minutes, every Mus auris dextra coating dimethylbenzene 0.05ml causes scorching moulding, after 4 hours, mice is pulled cervical vertebra puts to death, cut two ears along the auricle baseline, lay the ear disk at same position respectively, weigh with precision torsion balance with diameter 8mm card punch, deducting left auricle weight with every Mus auris dextra sheet is the swelling degree, and the swelling degree of matched group and administration group is compared and statistical procedures.Result such as table 6.

Table 6 medicine medicated powder of the present invention xylol causes the influence (X ± SD) of mice auricle swelling

Annotate: ^*: P＜0.05 (comparing) with matched group

Experimental result shows: medicine 0.5 of the present invention, and the 0.25g/kg xylol causes mice auricle swelling significant inhibitory effect, and relatively there were significant differences (P＜0.05) with matched group, and showing has significant resist inflammation on repercussive function.Its action intensity surpasses with the spirit of dosage larynx disease.

3, medicine on Carrageenan of the present invention causes the influence test of rat swollen feet

Get 60 of SD rats, be divided into 6 groups at random by body weight, a clear horizontal line is made with ball pen in upper end, the left back vola of every Mus front, survey rat foot moulding front volume and record with drainage, each organizes ig medicine 0.125 of the present invention respectively then, 0.25,0.5g/kg and larynx disease spirit 0.33g/kg and sc hydrocortisone 25mg/kg, matched group ig equal-volume distilled water.Draw the 1% chondrus ocellatus Holmes glue 0.1ml prepare with the 0.25ml syringe after 1 hour, it is subcutaneous to inject the left back foot sole of the foots of rat with No. 4 syringe needles, then respectively at measuring the left back sufficient volume of each rat 1 time, calculating Mus foot swelling percentage rate after the moulding in 1,2,4,6,8 hour.

Result such as table 7.

Table 7 medicine medicated powder of the present invention to the influence of rat swollen feet (X ± SD, n=10)

Annotate: ^*: P＜0.05 ^*: P＜0.01 ^{* *}: compare with matched group P＜0.001

Medicine of the present invention as can be seen from Table 7 causes the rat swollen feet to rat carrageenan inhibitory action, little, in, heavy dose of 0.125,0.25,0.5g/kg group all reduces than matched group swelling degree in 5 times measuring, in, small dose group had highly significant to reduce (P＜0.01) at 4,6,8 hours.Though the detumescence effect is lower than hydrocortisone,, show that medicine of the present invention has the bloated effect of detumescence preferably apparently higher than Chinese medicine contrast medicine larynx disease spirit group.

4, medicine of the present invention influence test that leukocyte in the carboxymethyl cellulose capsule is swum out of

Get 60 of SD rats, be divided into 6 groups, each group is ig medicine 0.125,0.25 of the present invention, 0.5g/kg respectively, larynx disease spirit 0.33g/kg and sc hydrocortisone 25mg/kg, matched group ig equal-volume distilled water.Once a day, for three days on end, meanwhile, back of the body cervical region lost hair or feathers with 5% sodium sulfide in the 2nd day, the subcutaneous injection of the cervical region 8ml air of under aseptic condition, supporting or opposing in the 3rd day, form round balloon, inferior daily sterile needle head and syringe only inject carboxymethyl cellulose (CMC) liquid 5ml/ in air bag, drew each 0.1ml of CMC liquid in the capsule in back 3 hours and 7.5 hours in injection, splash in the 3ml brilliant cresyl blue dye liquor, mixing dyeing was dripped on the WBC counting chamber after 2 minutes, in microscopically counting WBC, be converted into WBC sum in every capsule again, calculate WBC number in four jiaos of 4 big grids, according to formula (4 WBC sum ÷ 4 * 10 * 20 * 10 that grid is interior ⁶=WBC number/L).Result such as table 8.

The influence that table 8 medicine of the present invention is swum out of leukocyte in the carboxymethyl cellulose capsule (X ± SD)

Annotate: ^*: P＜0.05, ^{* *}: compare with matched group P＜0.001

Medicine medicated powder of the present invention as can be seen from Table 8 causes inflammation to the rat carboxymethyl cellulose and brings out the body leukocyte and swum out of certain inhibitory action, 3 dosage groups of medicine of the present invention all reduced than matched group WBC number in 2 times measuring, dosage group big or middle can significantly suppress leukocyte and assemble (P＜0.05) in scorching kitchen range at 7.5 hours, though the minimizing degree is not as good as hydrocortisone, but, show that medicine medicated powder of the present invention has the effect that suppresses the inflammatory reaction in mid-term apparently higher than Chinese medicine contrast medicine larynx disease spirit group.

5, medicine of the present invention is to the influence test of mice chronic inflammatory disease granuloma induced by implantation of cotton pellets

Get 72 of body weight 18～22g mices, male and female half and half.At every Mus chest iodine disinfection, after 75% cotton ball soaked in alcohol takes off iodine, cut an osculum at chest, with the ophthalmology tweezer 5mg autoclaving cotton balls is implanted subcutaneous axillary region, skin suture immediately from incision.Began administration the same day from performing the operation, 5 administration groups are distinguished ig medicine 1.0,0.5 of the present invention, 0.25g/kg, Chinese medicine matched group ig larynx disease spirit 0.66g/kg, western medicine group sc hydrocortisone 25mg/kg, blank ig equal-volume distilled water.Below respectively organize equal successive administration 6 days, weigh before the off-test in the 7th day, take off cervical vertebra then and put to death, will implant cotton balls and take out in the lump, reject fatty tissue, put into baking oven and dried 12 hours for 60 ℃, on precision torsion balance, weigh together with connective tissue on every side.To claim to such an extent that weight deducts the former weight of cotton balls and promptly gets granuloma weight, and calculate with mg/10gbw, and organize a comparison and statistical procedures.Result such as table 9.

Table 9 medicine of the present invention is to the influence of mice chronic inflammatory disease granuloma induced by implantation of cotton pellets (X ± SD)

Annotate: ^*: P＜0.05, ^*: P＜0.01, ^{* *}: compare with matched group P＜0.001

Mus death is arranged, number of animals when number of animals finishes for experiment in the table in the experimentation

Experimental result shows: medicine 1.0,0.5 of the present invention, and 0.25g/kg compares with matched group the mice granuloma induced by implantation of cotton pellets, and there were significant differences (P＜0.01 or P＜0.05), shows that 3 dosage of medicine of the present invention have obvious restraining chronic inflammation effect.The spirit of Chinese medicine contrast medicine larynx disease does not see that obvious effect is arranged.Medicine medicated powder antiinflammatory action of the present invention is better than with the spirit of dosage larynx disease.

6, medicine medicated powder of the present invention is to the influence test of rat chronic inflammation granuloma induced by implantation of cotton pellets

It is a collection of to get body weight 140～160g SD rat, is male, is divided into 6 groups at random.At every Mus chest iodine disinfection, after 75% cotton ball soaked in alcohol takes off iodine, cut an osculum at chest, with the ophthalmology tweezer 20mg autoclaving cotton balls is implanted subcutaneous axillary region, skin suture immediately from incision.Began administration the same day from performing the operation, 5 administration groups, difference ig medicine 0.5,0.25 of the present invention, 0.125g/kg; Chinese medicine matched group ig larynx disease spirit 0.33g/kg, western medicine group sc hydrocortisone 12.5mg/kg, blank group ig equal-volume distilled water.Below respectively organize equal successive administration 6 days, weigh once before experiment in the 7th day finishes, take off cervical vertebra then and put to death, to implant cotton balls takes out in the lump together with connective tissue on every side, reject fatty tissue, put into 60 ℃ of oven dry of baking oven 12 hours, on precision torsion balance, weigh up gross weight again.To claim to such an extent that weight deducts the former weight of cotton balls and promptly gets granuloma weight, and calculate with mg/100gBW, and organize a comparison and statistical procedures.Result such as table 10.

Table 10 medicine of the present invention is to the influence of rat chronic inflammation granuloma induced by implantation of cotton pellets (X ± SD)

: compare with matched group P＜0.05

Experimental result shows: medicine medicated powder 0.5g/kg of the present invention is to the swollen significant difference (P＜0.05) of having compared with matched group of rat granuloma, show that medicine medicated powder 0.5g/kg of the present invention has obvious restraining chronic inflammation effect, and 0.25g/kg and matched group relatively have and necessarily dwindle, but no difference of science of statistics, show that all the other dosage group restraining chronic inflammation effects are not obvious, Chinese medicine contrast medicine larynx disease spirit group does not see that obvious effect is arranged yet.Medicine medicated powder action intensity of the present invention surpasses the spirit of larynx disease.

The 6 medicine analgesic activity of the present invention experimental studies of test example

1, medicine Dichlorodiphenyl Acetate of the present invention causes the influence test (writhing method) of mice pain effect

Get 60 of body weight 18～22g mices, male and female half and half are divided into 6 groups at random, 3 administration group difference ig medicines 1.0,0.5 of the present invention, 0.25g/kg; Chinese medicine contrast medicine JINSANGZIHOUBAO 3.0g/kg, administration after 60 minutes every Mus ip 0.5%HAC 0.2ml/ only observe in 15 minutes and turn round the body number of times.Western medicine contrast medicine sc morphine hydrochloride 10mg/kg only inject after 20 minutes ip HAC0.2ml/, turns round the body number of times with the method observation in 15 minutes, and the result is compared statistics.Result such as table 12.

Table 12 medicine medicated powder of the present invention is to the analgesic activity (writhing method) of mice (X ± SD)

Annotate: ^*: P＜0.05 ^*: P＜0.01 ^{* *}: P＜0.001 (comparing) with matched group

Experimental result shows: medicine medicated powder 1.0g/kg of the present invention, 0.5g/kg that suppresses very significantly that acetic acid causes mice pain turns round the body number of times, with matched group highly significant significant difference (P＜0.01) is arranged relatively, illustrate that medicine medicated powder of the present invention has the analgesic activity of highly significant; JINSANGZIHOUBAO 3.0g/kg also significantly suppresses mouse writhing number of times (P＜0.05); The morphine hydrochloride subcutaneous injection then extremely significantly suppresses mouse writhing number of times (P＜0.001), shows that 3 kinds of medicines all have analgesic activity (chemical method causes pain, writhing method), and medicine medicated powder analgesic activity of the present invention is inferior to morphine, but the strong JINSANGZIHOUBAO of crossing.

2, medicine of the present invention causes the influence test (hot plate method) of mice pain effect to hot plate

The thermostatic water-circulator bath device is adjusted to 55 ± 0.5 ℃, made the hot plate preheating 10 minutes.

At first screen the qualified mice of temperature-sensitive: it is a collection of to get 18～22g female mice, puts 1 on hot plate at every turn, and mice is from being placed on the hot plate to licking the pain threshold of metapedes required time (second) as this Mus.All threshold values were less than 5 seconds or give it up greater than 30 seconds or leaper.60 of qualified mices are divided into 6 groups at random, repeat to survey its normal pain threshold, get twice pain threshold meansigma methods as this Mus administration before pain threshold.Administration then, 3 are subjected to reagent group ig medicine 0.25,0.5 of the present invention respectively, 1.0g/kg; Chinese medicine contrast medicine group ig larynx disease spirit 0.66g/kg and Western medicine contrast medicine group sc morphine hydrochloride 10mg/kg; Blank group ig equal-volume distilled water.Respectively at after the administration 15 minutes, 30 minutes, 60 minutes, measured the pain threshold of each mice in 90 minutes.Experiment finishes, and each is organized each time average pain threshold compare, and processing takes statistics.Result such as table 13.

Table 13 medicine medicated powder of the present invention is to the analgesic activity (hot plate method) of mice

Annotate: ^*: P＜0.05 ^*: P＜0.01 (comparing) with matched group

Experimental result shows: the pain threshold of each group is improved after the administration, and is the most outstanding with the morphine hydrochloride group, and each minute is all than there being highly significant to improve (P＜0.01) before the administration; Medicine medicated powder of the present invention, most of minute pain threshold all have before than administration more obviously and improve, and big or middle dosage measured in 90 minutes that contrast is significantly increased (P＜0.05) after administration, show that medicine medicated powder of the present invention has significant analgesic activity to thermostimulation.

The 7 medicine refrigeration function tests of the present invention of test example

Get 80 of rats, survey anus temperature 2 time (upper and lower noon respectively once) with mercury clinical thermometer every day, continuous 2 days, the same day was surveyed body temperature 2 times in test again, chose 60 of the rats that the body temperature fluctuation is no more than 0.3 ℃, be divided into 6 groups at random, every group 10, each group is ig medicine 0.125,0.25 of the present invention respectively, 0.5g/kg with larynx disease spirit 0.33g/kg and aspirin 25mg/kg, matched group ig equal-volume distilled water.Each 0.1ml/ of injection 1% chondrus ocellatus Holmes glue is sufficient down in the two metapedes two sufficient sole of the foot mesotheliums of rat simultaneously in administration, respectively surveys the anus temperature once respectively at 2,4,6,8,10 hours then, obtains the meansigma methods that each minute body temperature changes, and relatively, the result adds up between organizing.Result such as table 14.

Table 14 medicine medicated powder of the present invention is to the refrigeration function of rat carrageenan pyrogenicity

Annotate: ^*: P＜0.05 (comparing) with matched group

Medicine medicated powder of the present invention as can be seen from Table 14 has certain refrigeration function to rat carrageenan pyrogenicity, large, medium and small dosage 0.5,0.25,3 dosage groups of 0.125g/kg group all are lower than matched group in the body temperature meansigma methods of most minutes, 6,8,10 hours body temperature of large and small dosage group reduces more obvious than matched group, obviously be better than Chinese medicine contrast medicine larynx disease spirit group, and heavy dose of 10 hours significant differences (P＜0.05) show that medicine medicated powder of the present invention has certain refrigeration function.

The antitussive action test of test example 8 medicine medicated powder of the present invention

60 of acquisition Kunming mouses, ♀, ♂ half and half, under Traditional Chinese Medicine Research Institute, Sichuan Province cleaning level animal housing experiment condition, conformed 24 hours, divide 6 groups then, that is: the same volume of normal control group (with the same volume of administration group) distilled water, medicine 1.0,0.5 of the present invention, 0.25g/kg 3 dosage groups, Chinese medicine contrast medicine JIZHI TANGJIANG 20ml/kg group, Western medicine contrast medicine ig codeine phosphate 30mg/kg, each organizes equal successive administration 3 days, last administration 40 minutes, respectively mice is placed in the inverted 500ml beaker, inject 0.15ml/ strong aqua ammonia in the cuvette of in beaker, placing, smoked for 15 seconds through ammonia, take out mice, count 2 fens interior cough number of times of clock time of mice, and the result is carried out statistical disposition, relatively make the significance of difference between group and measure.Result such as table 15.

Table 15 medicine medicated powder of the present invention draws the antitussive action of coughing mice (X ± SD) to strong aqua ammonia

The result shows: codeine group cough number of times reduces P (＜0.01) than the matched group highly significant, and JIZHI TANGJIANG group also comparative control group significantly reduces (P＜0.05), shows Western medicine contrast medicine, and Chinese medicine contrast medicine all has remarkable antitussive action; The cough number of times of 3 dosage groups of medicine of the present invention all reduces than matched group, and there were significant differences (P＜0.05) for heavy dose of group, shows that medicine medicated powder heavy dose of the present invention has remarkable antitussive action.

Prove by the multiple test of pesticide effectiveness:

1. medicine medicated powder of the present invention has significant antivirus action in vitro tests, medicine medicated powder 10 of the present invention, 5,2.5mg/ml can suppress Coxsackie virus (CoxB) 3 cytopathogenic effects fully, with consistent with the effect of dosage Andrographolide, and virazole has only 10mg/ml that this effect is arranged, 5.0, and 2.5mg/ml fails to suppress (CoxB) 3 cytopathogenic effects; Medicine medicated powder 10mg/ml of the present invention can suppress adenovirus (AdV) 3 type cytopathogenic effects, and is consistent with the virazole action intensity with dosage.And this concentration of Andrographolide does not show the effect that suppresses adenovirus.

Medicine medicated powder 10 of the present invention, 5mg/ml can suppress respiratory syncytial virus (RSV) cytopathogenic effect fully, this effect is weaker than virazole (virazole 0.5mg/ml can suppress RSV), consistent with the Andrographolide effect, can suppress influenza virus A 1 type cytopathogenic effect, the consistent strong effect (virazole only just has this effect when 10mg/ml) of crossing virazole of this effect with the effect of Andrographolide; Medicine medicated powder 10mg/ml of the present invention can suppress influenza virus A 3 type cytopathogenic effects, this action intensity is but consistent with the effect of virazole not as good as Andrographolide (Andrographolide 5mg/ml promptly have this effect), and all being has inhibition can suppress influenza virus A 3 type cytopathic effects in concentration during for 10mg/ml.

2. medicine 1.0,0.5 of the present invention, 0.25g/kg causes the inhibitory action that the mouse peritoneal capillary permeability increases highly significant to lumbar injection acetic acid, and showing has the anti-inflammatory of highly significant transudation, and its action intensity is than strong with the dosage watermelon crystal.

3. medicine 0.5 of the present invention, the 0.25g/kg xylol causes mice ear significant inhibitory effect, and showing has significant resist inflammation on repercussive function, and action intensity far surpasses with the spirit of dosage larynx disease.

4. medicine medicated powder 0.5,0.25 of the present invention, the 0.125g/kg on Carrageenan causes the inhibitory action that the rat swollen feet has highly significant, shows that medicine of the present invention has good resist inflammation on repercussive function, and action intensity is lower than hydrocortisone, is significantly higher than the spirit of larynx disease.

5. medicine medicated powder 0.5 of the present invention, 0.25,0.125g/kg being brought out the body leukocyte, carboxymethyl cellulose swum out of certain inhibitory action, dosage group big or middle can significantly suppress leukoplania (P＜0.05) at 7.5 hours, show that medicine of the present invention has the effect of inflammation-inhibiting interim response, action intensity surpasses the spirit of larynx disease, not as good as hydrocortisone.

6. medicine medicated powder 1.0,0.5 of the present invention, large, medium and small 3 the dosage groups of 0.25g/kg all have significant inhibitory effect to the mice granuloma induced by implantation of cotton pellets, show that medicine of the present invention has the restraining chronic inflammation effect, and action intensity surpasses the spirit of larynx disease.

7. medicine medicated powder 0.5g/kg of the present invention has remarkable inhibitory action to rat granuloma is swollen, shows that medicine of the present invention has the restraining chronic inflammation effect, and action intensity surpasses the spirit of larynx disease.

8. what medicine medicated powder 1.0 of the present invention, 0.5g/kg suppressed very significantly that acetic acid causes mice pain turns round the body number of times, shows that medicine of the present invention causes the analgesic activity that pain has highly significant to chemical agent, and its intensity is lower than morphine, surpasses JINSANGZIHOUBAO.

9. medicine medicated powder 1.0,0.5 of the present invention, 0.25g/kg has significant analgesic activity to the mice thermostimulation, and it is suitable with larynx disease spirit effect that intensity is lower than morphine.

10. medicine medicated powder 0.5,0.25 of the present invention, 0.125g/kg on Carrageenan pyrogenicity rat has certain refrigeration function, and intensity surpasses the spirit of larynx disease.

11. medicine medicated powder 1.0g/kg of the present invention draws the mice that coughs to strong aqua ammonia significant antitussive action is arranged.

Conclusion: medicine of the present invention has significant antivirus action, can significantly suppress the in-vitro multiplication of Coxsackie virus (CoxB3) and adenovirus (AdV3), and is comprehensive than the effect of Western medicine antiviral agents virazole and Chinese medicine antiviral agents Andrographolide; Medicine of the present invention has significant antiinflammation, and the acute inflammation that can suppress animal significantly oozes out, swelling (mice ear, rat paw edema), can suppress inflammation in mid-term (leukocyte is swum out of) and chronic inflammatory disease (large and small Mus granuloma) significantly; Medicine of the present invention has significant analgesic activity, and chemical agent is caused pain, thermic all analgesic activity bitterly; Medicine of the present invention has tangible refrigeration function and antitussive action.

In sum, medicine energy antiviral of the present invention, antiinflammatory, analgesia, analgesic, cough-relieving, determined curative effect, rapid-action, safety, acute pharyngolaryngitis or upper respiratory tract infection that all kinds of factors such as flu, excessively smoking, chemical irritant are caused present laryngopharynx swelling and pain, hoarseness, the headache fever, cough expectorant Huang or symptoms such as throat, nasal mucosa hyperemia have the obvious treatment effect, and cardinal symptom can be fully recovered at 3～7 days.Treat the medicine of acute and chronic pharyngolaryngitis.The preparation method of medicine of the present invention is that separation and purification goes out the effective site total flavones from primary crude drug, adopt macroreticular resin absorbing method, technology advanced person, cost is low, the efficient height, the no three wastes, and added value height, profit margin is big, primary crude drug Rhizoma Iridis Tectori cheap at present (4～5 yuan/kilogram), the yield that adopts extracting method purification total flavones of the present invention is about 15%, just the per kilogram primary crude drug can prepare 400 of medicine capsules of the present invention (0.35g/ grain), the direct production cost of every capsules is about 0.07 yuan/, medical material per ton can be created 280,000 yuan of the output values, if selling cost is 40% of an income from sales, the product that medical material per ton is produced can obtain taxes and profits 28-28 * 40%-2.8 (production cost)=140,000 yuan, and promptly long-pending profit is 50% of an income from sales.If 250 tons of primary crude drugs of annual processing, taxes and profits promptly have 3,500 ten thousand yuan (output is 100,000,000, and the output value and income from sales are 7,000 ten thousand), this shows, medicine of the present invention is in commercial achieving success, be owing to having selected primary crude drug cheaply, under material medicine extracting method of the present invention, the yield height, directly cause by technical characterictic of the present invention, for a person skilled in the art, be non-obvious, provide a kind of new medicinal selection for clinical.

Claims (10)

1, the purposes of irides Rhizoma Iridis Tectori Iris tectorum Maxim. extractive of general flavone in the preparation antiviral drugs, wherein, the percentage composition of total flavones is with iridin C in the extractive of general flavone ₂₂H ₂₂O ₁₁Calculate 〉=55%.

2, purposes according to claim 1 is characterized in that: contain in the described total flavones: iridin tectoridin, iristectorin A iristectorin A, iris aglycone tectorigenin, wild flag aglycon irigenin

3, purposes according to claim 2 is characterized in that: the weight proportion iridin of four isoflavones components in the total flavones: iristectorin A: iris aglycone: the wild flag aglycon is 1: 0.1～0.4: 0.15～0.6: 0.03～0.1.

4, purposes according to claim 3 is characterized in that: the content of four isoflavones components is than iridin in the total flavones: iristectorin A: iris aglycone: the wild flag aglycon is 1: 0.18: 0.29: 0.056.

5, purposes according to claim 1 is characterized in that: described Rhizoma Iridis Tectori extractive of general flavone is by following method preparation:

A, get irides Rhizoma Iridis Tectori Iris tectorum Maxim. medical material, pulverize, add 70% alcohol reflux, concentrate, extractum;

In b, the extractum dissolving entry with a step, filter, filtrate is passed through macroporous adsorptive resins;

C, wash the fat post with water, effluent discards, and when effluent is nearly achromatism and clarity solution, puts solid carbon dioxide, the ethanol elution with 75～90%, and eluent filters, and concentrates promptly to get extractive of general flavone.

6, purposes according to claim 5 is characterized in that: the weight proportion of the amount of resin of the described macroporous adsorptive resins of b step and primary crude drug is in the preparation method of described Rhizoma Iridis Tectori extractive of general flavone: 3: 1.

7, purposes according to claim 1 is characterized in that: the dosage form of described antiviral drugs is: capsule, granule, tablet, pill or oral liquid.

8, purposes according to claim 1 is characterized in that: the preparation method of described antiviral drugs comprises the steps:

A, get irides Rhizoma Iridis Tectori Iris tectorum Maxim. medical material, pulverize, add 70% alcohol reflux, concentrate, extractum;

In b, the extractum dissolving entry with a step, filter, filtrate is passed through macroporous adsorptive resins;

C, wash the fat post with water, effluent discards, and when effluent is nearly achromatism and clarity solution, puts solid carbon dioxide, and with 75～90% ethanol elutions, eluent filters, and concentrates and promptly gets total flavones;

D, take by weighing the total flavones of c step, add acceptable accessories or complementary composition and be prepared into preparation pharmaceutically commonly used.

9, purposes according to claim 1 is characterized in that: described antiviral drugs is resisiting influenza virus, parainfluenza virus, adenovirus, its virus of Ke's Sa, respiratory syncytial virus, rhinovirus or herpesvirus medicament.

10, the purposes of irides Rhizoma Iridis Tectori Iris tectorum Maxim. extractive of general flavone in the preparation analgesic, wherein, the percentage composition of total flavones is with iridin C in the extractive of general flavone ₂₂H ₂₂O ₁₁Calculate 〉=55%.