CN101879214B

CN101879214B - Chinese medicinal composition for treating bronchial asthma and preparation method thereof

Info

Publication number: CN101879214B
Application number: CN200910136303A
Authority: CN
Inventors: 卢印英
Original assignee: BEIJING JIABO HONGKANG PHARMACEUTICAL TECHNOLOGY Co Ltd
Current assignee: BEIJING JIABO HONGKANG PHARMACEUTICAL TECHNOLOGY Co Ltd
Priority date: 2009-05-08
Filing date: 2009-05-08
Publication date: 2012-09-26
Anticipated expiration: 2029-05-08
Also published as: CN101879214A

Abstract

The invention discloses a Chinese medicinal composition for treating bronchial asthma and a preparation method thereof. The Chinese medicinal composition comprises the following raw material medicaments in part by weight: 5 to 20 parts of pungent oleaster leaf, 4 to 18 parts of epimedium brevicornum and 2 to 12 parts of Chinese magnoliavine fruit. Capsules, tablets, granules, gels, slow-release formulations, oral liquid or pills are prepared by extracting pungent oleaster leaf, the epimedium brevicornum and the Chinese magnoliavine fruit through a conventional process method and adding pharmaceutically-acceptable auxiliary materials into the extract. The Chinese medicinal composition has the excellent effects of relieving asthma and stopping cough and reinvigorating and consolidating the constitution.

Description

A kind of Chinese medicine composition of treating bronchial asthma and preparation method thereof

Technical field

The present invention relates to a kind of Chinese medicine composition and preparation method thereof, particularly a kind of Chinese medicine composition of treating bronchial asthma and preparation method thereof.

Background technology

Bronchial asthma (Bronchial athma) is to be clinical common, the refractory of main performance, anxious critical illness disease with the bronchospasm, belongs to the traditional Chinese medical science (breathing heavily) the sick category of roaring.Recently add up according to World Health Organization (WHO), the whole world has 1.6 hundred million patients approximately, and the prevalence of U.S.'s asthma is up to 4%, and Japan is 3%, Italian rural area 5%, and city 7%, China is about 2%, and the whole nation has more than 2,000 ten thousand patients approximately.Along with industrial expansion, atmospheric pollution serious, the living environment allergen increases, and the pathogenesis of asthma rate rises, and old people's asthma prevalence is also increasing.January calendar year 2001; World Health Organization (WHO) just once delivered " global strategy of asthma control ", because this disease is shown effect repeatedly, onset is anxious; Be more common in child and person between twenty and fifty; And be prone to cause emphysema, pulmonary heart disease, cause disability and viability, also classify it as common refractory anxious critical illness in China.

From the doctor trained in Western medicine angle: bronchial asthma (bronchial wheezing) is the air flue chronic inflammation illness of being participated in by various kinds of cell (like eosinophilic granulocyte, mastocyte, T lymphocyte, neutrophilic granulocyte, airway epithelia cell etc.) and cell component (cellularelements).This chronic inflammatory disease causes the increase of airway hyperreactivity; Extensively changeable reversibility flow limitation appears usually; And cause symptoms such as the panting of repeated relapsing, out of breath, uncomfortable in chest or cough be everlasting night and (or) outbreak in early morning, aggravation, but most of patients spontaneous remission or alleviate through treatment.Doctor trained in Western medicine is removed bronchus convulsion medicine such as theophylline class medicine: (therapeutic dose 10-20mg/ml and toxic dose 20mg/ml are very approaching for aminophylline; Be prone to toxic reaction; Make medicament blood concentration monitor control consumption clinically; And the side effect of cardiovascular aspect arranged), receptor, analeptic (epinephrine, ephedrine etc.; Only be used to not have hypertension and cardiovascular disease person), epinephrine corticosteroids (cause iatrogenic gastric ulcer, hypertension, water-sodium retention, athero, diabetes etc., and be easy to generate rely on and opposing) etc.

The argumentation beginning of the relevant bronchial wheezing of the traditional Chinese medical science is shown in Huangdi's Internal Classics; The Song dynasty king is fair and unbiased will to roar first and separate with breathing heavily, and not discuss but have in detail, just begin clearly asthma as independent name of disease until Yuan Dynasty Zhu Dan small stream; " heavy breathing " card and " breathing heavily " card separate argumentation and are derived from the Ming Dynasty and deceive and to fight.Heterogeneous, the insufficiency of primordial QI of generation and natural endowment of bronchial wheezing, exopathogen invasion and attack, injury due to diet, weak after being ill, positive QI-insufficiency is disturbed in the pathogen, mechanism of qi is inappropriate, the five internal organs pathological changes, involve in factors such as lungs relevant.

The Chinese medicine that is used to remove bronchospasm does not at present still have efficient target effect, still continues to use traditional diagnosis and prescription.The Chinese medicine compound late result is remarkable, but prescription is big, and onset is slow, takes inconvenience.The single medicinal material preparation (like Cuculus polioephalus, Herba Viticis Cannabifoliae, Flos Daturae etc.) of exploitation has preferably the curative effect of eliminating the phlegm, relieving asthma clinically in recent years, is main to eliminate the phlegm, though relieving asthma effect still can't with many side effect are arranged, but comparatively fast the Western medicine of relief of symptoms is compared.

Therefore, the seek treatment active drug of bronchial asthma (bronchial wheezing) is pressing for of clinical practice.

Summary of the invention

One object of the present invention is to disclose a kind of Chinese medicine composition of treating bronchial asthma;

Another object of the present invention is to disclose a kind of method for preparing of treating the Chinese medicine composition of bronchial asthma;

Another object of the present invention is to disclose a kind of Chinese medicine composition drop pill of treating bronchial asthma and preparation method thereof.

The present invention seeks to realize through following technical scheme:

A kind of Chinese medicine composition of treating bronchial asthma of the present invention, its crude drug consists of (weight ratio):

Folium Elaeagni 5-20 weight portion, Herba Epimedii 4-18 weight portion, Fructus Schisandrae Chinensis 2-12 weight portion.

The Chinese medicine composition of above-mentioned treatment bronchial asthma, its crude drug composition is preferably:

Folium Elaeagni 9-15 weight portion, Herba Epimedii 6-12 weight portion, Fructus Schisandrae Chinensis 4-9 weight portion.

Folium Elaeagni 10 weight portions, Herba Epimedii 9 weight portions, Fructus Schisandrae Chinensis 5 weight portions.

Folium Elaeagni 14 weight portions, Herba Epimedii 9 weight portions, Fructus Schisandrae Chinensis 7 weight portions.

Folium Elaeagni 12 weight portions, Herba Epimedii 9 weight portions, Fructus Schisandrae Chinensis 6 weight portions.

Folium Elaeagni (Elaeagnus pungens Thunb.) is the leaf of Elaeangnaceae oleaster Fructus Elaeagni.Above-mentioned each crude drug all can the proximate medicine replacement of selection function from " Chinese ministry standard ", " pharmacopeia " or " Chinese medicine dictionary " etc.

The present invention also provides the method for preparing of the Chinese medicinal composition preparation of above-mentioned treatment bronchial asthma; This method is: Folium Elaeagni, Herba Epimedii, Fructus Schisandrae Chinensis add the pharmacy acceptable auxiliary according to the above ratio and process the acceptable any conventional dosage form of pharmaceutics after conventional process is extracted, comprise capsule, tablet, granule, gel, slow releasing agent, oral liquid or drop pill; Said common process method comprises water extraction or alcohol extraction or first water extraction water extraction again after alcohol extraction or the first alcohol extraction again, and it is further active constituent-enriched after above-mentioned conventional the extraction, can also to cross macroporous resin column;

Above-mentioned common process water extracting method can for: each 10-30 doubly measures, and extracts 1-3 hour at every turn 1-4 time;

Above-mentioned common process alcohol extraction method can for: 30-80% ethanol, 4-15 doubly measured, extract respectively 1-3 hour;

The said macroporous resin column method of crossing is: extracting solution filters, and filtrating is through macroporous resin, and elder generation is with the ethanol elution of 4-6 times of water gaging or 20% following concentration; Eluent is abandoned or adopted, and reuse adds 1-10 times of eluting of 50-80% ethanol, collects eluent; Decompression recycling ethanol gets the purification extracting solution to there not being the alcohol flavor.

The present invention also further provides the method for optimizing of preparation of the Chinese medicinal composition preparation of above-mentioned treatment bronchial asthma, and this method is:

The extraction of A, Folium Elaeagni effective ingredient

A1: the Folium Elaeagni water boiling and extraction, each 8-12 doubly measures, 0.5-2 hour, extracts 2-4 time; Merge extractive liquid, filters, and filtrating concentrates; Adopt the macroporous resin enrichment effective ingredient, the water elution remove impurity uses 50-70% ethanol to be the eluant eluting; Behind the eluent decompression recycling ethanol, drying under reduced pressure is pulverized to fine powder subsequent usely, is Folium Elaeagni extract I; Or

The 50-70% ethanol extraction that A2:7-10 doubly measures 2-3 time extracted respectively 1-3 hour, and merge extractive liquid, filters, and decompression filtrate recycling ethanol is concentrated subsequent use, is Folium Elaeagni extract II;

The extraction of B, Fructus Schisandrae Chinensis effective ingredient: Fructus Schisandrae Chinensis 7-10 doubly measures the 40-60% ethanol extraction 2-3 time, extracts respectively 1-3 hour, and merge extractive liquid, filters, and decompression filtrate recycling ethanol concentrates subsequent use;

The extraction of C, epimedium active constituent: Folium Epimedii adds the water 20-40 times water yield at every turn, decocts to extract 2-4 time each 1-3 hour; Extracting solution filters, and merging filtrate concentrates; Adopt the macroporous resin enrichment effective ingredient, the water elution volume is 6-12BV; Eluting solvent is a 60-80% ethanol; Behind the eluent decompression recycling ethanol, drying under reduced pressure, pulverize into fine powder subsequent use;

With above-mentioned Folium Elaeagni extract I or II; Or I and II and Fructus Schisandrae Chinensis extrat, Herba Epimedii extract mix homogeneously; Add the pharmacy acceptable auxiliary and process the acceptable any conventional dosage form of pharmaceutics, comprise capsule, tablet, granule, gel, slow releasing agent, oral liquid or drop pill; In the situation of said Folium Elaeagni extract I and II and Fructus Schisandrae Chinensis extrat, Herba Epimedii extract mix homogeneously; Folium Elaeagni extract I is made by in the Folium Elaeagni raw material recipe quantity half, and Folium Elaeagni extract II is made by in the Folium Elaeagni raw material recipe quantity second half.

The present invention also further provides the most preferred method of preparation of the Chinese medicinal composition preparation of above-mentioned treatment bronchial asthma, and this method is:

The extraction of A, Folium Elaeagni effective ingredient

A1: the Folium Elaeagni water boiling and extraction, each 10 times of amounts, 1 hour are extracted 3 times; Merge extractive liquid, filters, and filtrating is concentrated into 1: 1, and (medical material: filtrating is g: ml); Adopt the macroporous resin enrichment effective ingredient, the sample solution concentration of extract is 0.1g crude drug/ml, appearance on the speed of 2BV/h; The water elution remove impurity of 4BV/h uses 60% ethanol to be the eluant eluting, and elution flow rate is that 3BV/h, elution volume are 4BV; Behind the eluent decompression recycling ethanol, drying under reduced pressure is pulverized to fine powder subsequent usely, is Folium Elaeagni extract I; Or

60% ethanol extraction that A2:8 doubly measures extracted respectively 1.5 hours, 1.5 hours, and merge extractive liquid, filters, and decompression filtrate recycling ethanol is concentrated into relative density and is 1.15-1.25 (50 ℃), and is subsequent use, is Folium Elaeagni extract II.

The extraction of B, Fructus Schisandrae Chinensis effective ingredient

After Fructus Schisandrae Chinensis is broken for coarse granule, 8 times of amount 50% ethanol, extracted 1.5 hours, 1.5 hours respectively, merge extractive liquid, filters, and decompression filtrate recycling ethanol is concentrated into relative density and is 1.15-1.25 (50 ℃), and is subsequent use.

The extraction of C, epimedium active constituent

Folium Epimedii adds 30 times of water yields of water at every turn, decocts to extract each 1.5 hours 3 times.Extracting solution filters, and merging filtrate is condensed into 1: 1 (medical material g: filtrating ml); Select the rich composition of imitating of macroporous resin for use, sample solution concentration is 0.5g crude drug/ml; Applied sample amount is 1g crude drug/ml resin; Adsorption rate is 0.5ml/min; Elution speed is 1.0ml/min; The water elution volume is 10BV; Eluting solvent is 70% ethanol, and consumption is 6BV.Behind the eluent decompression recycling ethanol, drying under reduced pressure, pulverize into fine powder subsequent use;

Above-mentioned macroporous resin column can be selected from AB-8 resin, D-101 resin, HPD-100 or HPD-450 resin;

" doubly measure " weight ratio of the crude drug that being meant compares is extracted described in the consumption of said extracted liquid.

Said pharmacy acceptable auxiliary is: filler, disintegrating agent, lubricant, suspending agent, binding agent, sweeting agent, correctives, antiseptic, substrate etc.Filler comprises: starch, pregelatinized Starch, lactose, mannitol, chitin, microcrystalline Cellulose, sucrose etc.; Disintegrating agent comprises: starch, pregelatinized Starch, microcrystalline Cellulose, carboxymethyl starch sodium, crospolyvinylpyrrolidone, low-substituted hydroxypropyl cellulose, cross-linking sodium carboxymethyl cellulose etc.; Lubricant comprises: magnesium stearate, sodium lauryl sulphate, Pulvis Talci, silicon dioxide etc.; Suspending agent comprises: polyvinylpyrrolidone, microcrystalline Cellulose, sucrose, agar, hydroxypropyl emthylcellulose etc.; Binding agent comprises, starch slurry, polyvinylpyrrolidone, hydroxypropyl emthylcellulose etc.; Sweeting agent comprises: saccharin sodium, Aspartane, sucrose, cyclamate, enoxolone etc.; Correctives comprises: sweeting agent and various essence; Antiseptic comprises: parabens, benzoic acid, sodium benzoate, sorbic acid and its esters, benzalkonium bromide, fixed, the Folium eucalypti globueli (Eucalyptus globulus Labill.) wet goods of acetic acid chloroethene; Substrate comprises: PEG6000, PEG4000, insect wax etc.

The present invention also provides a kind of Chinese medicine composition drop pill of treating bronchial asthma, in this drop pill:

Crude drug (Folium Elaeagni, Herba Epimedii, Fructus Schisandrae Chinensis) extract is 20-33%, and substrate is PEG4000 and PEG6000, PEG4000: PEG6000=6-11: 1; Wherein drug loading is preferred 30%, PEG4000: PEG6000=8: 1; Said extract both can be that Folium Elaeagni, Herba Epimedii, Fructus Schisandrae Chinensis close and carry thing, was preferably above-mentioned Folium Elaeagni extract I or II, or I and II mix with Fructus Schisandrae Chinensis extrat, Herba Epimedii extract.

The present invention also provides a kind of method for preparing of treating the Chinese medicine composition drop pill of bronchial asthma, and this method is:

With the drug loading of 20-33% add substrate (PEG4000: PEG6000=6-11: 1), mixing, condensation temperature is controlled at 2 ℃ 6 ℃; Dripping the system condition is: drip 70-75 ℃ of system temperature, drip apart from 4cm, cooling column length 75cm; The preparation drop pill.

In the invention described above compositions except the form that feeds intake with crude drug; Can also be directly feed intake with the form of extract; Concrete proportionate relationship can convert according to the crude drug inventory; Folium Elaeagni extract, Herba Epimedii extract, Fructus Schisandrae Chinensis extrat can also can reach the method preparation of prior art instruction according to the method described above for commercially available; Wherein extract is that alcohol extraction is crossed the refining thing of post, water and proposed any one or a few the combination in post refining thing, ethanol extract or the water extract.

The bronchial asthma that the present composition is controlled (bronchial wheezing) is because of due to the lung kidney deficiency, belongs to deficiency in origin and excess in superficiality, and deficiency in origin is master's a card; The not main gas of lung, failure of the kidney in receiving QI have caused the lasting difficulty of bronchial wheezing to heal.Folium Elaeagni meridian distribution of property and flavor in the side: sour, little hardship, return lung meridian; Be longer than lung cold cough with asthma or chronic cough of deficiency lung, asthma card.Folium Elaeagni is astringed the lung in the prescription, and antiasthmatic-antitussive uses and is monarch drug; Herba Epimedii (Herba Epimedii) meridian distribution of property and flavor: Xin Wen returns the Liver and kidney warp; The kind yang deficiency card of breathing with cough of controlling.Herba Epimedii kidney invigorating and YANG supporting in the prescription, antiasthmatic-antitussive, eliminating the phlegm to use is ministerial drug.The Fructus Schisandrae Chinensis temperature; Sour, sweet; Return lung, the heart, kidney channel; Restrain astringent or styptic treatment for spontaneous sweating, supplementing QI for promoting the production of body fluid, kidney calming; Be used for the chronic cough dyspnea due to deficiency, the deficient pulse etc. of losing heart.Fructus Schisandrae Chinensis is received contrary gas and is pacified lung and also use and be ministerial drug in the prescription.Three medicine the monarch and his subjects mutual reinforcements between are for using.Full side becomes the effect of antiasthmatic-antitussive, element culturing fixed folder altogether.

The relieving asthma active component of drop pill (product of embodiment 1 preparation) Chinese medicine of celestial spirit, effective site is clear and definite.Determined curative effect in clinical practice, and have antibiotic, antivirus action, the excitation heart is not found obvious toxic and side effects.

Irritate stomach respectively and give mice present composition preparation 0.39g/kg, 0.78g/kg and 1.17g/kg; The celestial spirit of Cavia porcellus 0.27g/kg, 0.54g/kg and the 1.08g/kg drop pill of relievining asthma; Can obviously prolong and cough the cough of coughing Cavia porcellus due to mice, the citric acid due to the ammonia and incubation period occurs; Reduce the cough number of times, show tangible antitussive effect; Obviously increase mouse bronchial phenol red output, show tangible phlegm-dispelling functions, Cavia porcellus due to the histamine phosphate " is breathed heavily heavys breathing ", and outbreak also has the obvious suppression effect, and prolongation is panted and twitched incubation period, shows tangible antiasthmatic effect; Rat paw edema due to acute ear swelling of Oleum Tiglii induced mice and the carrageenin also there is certain inhibitory action, explains that present composition preparation has the effect of certain anti-acute exudative inflammatory swelling; External bacteriostatic experiment is observed present composition preparation all has tangible bacteriostasis to respiratory tract common pathogens such as staphylococcus aureus, streptococcus pneumoniae, beta hemolytic streptococcuss; Laboratory observation obviously reduces staphylococcus aureus and pneumococcal infection mortality of mice to present composition preparation in the body, explains that present composition preparation has bacteriostasis in the tangible body.

Clinical research is the result show: estimate treatment group " celestial spirit relieving asthma prescription " treatment chronic duration of bronchial asthma (traditional Chinese medical science asthma, genus lung kidney deficiency type) 30 routine patients with multinomial symptom, sign (cough, cough up phlegm, pant, wheezing sound); Face control person's 11 examples; Produce effects person's 11 examples, responder's 7 examples, nonresponder's 1 example; Show control rate 73.34%, effective percentage 93.34%; Matched group " YUPINGFENG SAN closes seven flavor Duqi Wan prescriptions " treatment chronic duration of bronchial asthma (traditional Chinese medical science asthma belongs to lung kidney deficiency type) 10 routine patients face control person's 1 example, produce effects person's 4 examples, and responder's 3 examples, nonresponder's 2 examples show control rate 50%, effective percentage 80%.The clinical efficacy of " celestial spirit relieving asthma prescription " treatment chronic duration of bronchial asthma (traditional Chinese medical science asthma belongs to lung kidney deficiency type) is superior to the curative effect of matched group.In the improvement of symptom and sign, " celestial spirit relieving asthma prescription " is being superior to matched group aspect the wheezing sound relievining asthma to reduce, and the treatment group is having similar effect with the treatment group aspect the cough-relieving.Under observation have no adverse reaction.

Clinical experiment is the result show: lymphocyte transformation experiment and complement C3 change relatively before and after treatment group and the treatment of control group; Two groups of patients are after treatment; Lymphocyte transformation experiment and complement C3 content all have increasing in various degree; Handle P＜0.05 through statistics (t check), present remarkable meaning, explain that two groups of treatments all have immune defence potentiation.Compare zero difference (P＞0.05) between two groups, explain that tentatively the two has certain concordance.PGE before and after treatment group and the treatment of control group ₂, TXB ₂Change relatively treatment group treatment back PGE ₂Content increases, and there were significant differences to handle P＜0.05 through statistics (t check); Though matched group has increase, P＞0.05 not statistically significant.This explanation treatment group is being alleviated bronchospasm, and the reduction airway resistance is better than matched group with the effect of the aspect of relievining asthma.After two groups of treatments, TXB ₂Content all has decline in various degree, P＜0.05 significant difference, P between group＞0.05 zero difference.Show two groups reducing pulmonary vascular resistance, regulate the lung microcirculation, further improve the pulmonary function aspect and have concordance.IgG IgA IgM (enzyme linked immunosorbent assay mensuration) relatively before and after treatment group and the treatment of control group; Two groups of treatments all influence content of immunoglobulin IgG to some extent; Be after the treatment in various degree and reduce; Self compare P＜0.05 before and after two groups of treatments, there were significant differences on the statistics, explains that two groups of correlative factors to air flue spasm and obstruction have the positivity influence.Blood IgE IgA before and after treatment group and the treatment of control group, expectorant IgE IgA (putting the method for exempting from measures) compares, and after two groups of treatments, expectorant, blood IgA content improve, and expectorant, blood IgE content descend, through statistical procedures P＜0.05, P between group＞0.05.Show that tentatively two groups have positive role aspect the adjusting of the immune defence mechanism of lung.Treatment group and matched group are to pulmonary function (FEV _1.0, PEFR, FVC) improvement is in various degree all arranged, and the treatment group is more obvious.Through between two groups of treatment front and back of statistical procedures self and group, relatively all significant difference is arranged P＜0.05 through statistical procedures.

The product of drop pill of relievining asthma of celestial spirit described in the present invention for embodiment 1 preparation.Following experimental example and embodiment do further explanation to the present invention, but do not constitute restriction of the present invention.

Experimental example 1 pharmacodynamics test

Experiment material:

Trial drug: the celestial spirit drop pill of relievining asthma, every drop pill 0.05g, every pastille is equivalent to crude drug amount 0.45g, lot number: 010909, be compound Chinese medicinal preparation, be that Drug Manufacturing Room provide by Beijing University Of Chinese Medicine Traditional Chinese Medicine College's pharmacy.It is subsequent use by distilled water preparation to be made into 1.95%, 3.9%, 5.85% and 2.7%, 5.4%, 10.8% suspension (refer to drop pill holds separate thing) respectively.The dosage design considerations: each 20 of clinical adult, every day 3 times, one day totally 60, take into dose 3g every day, close to take every day and be equivalent to crude drug amount 27g.The dosage design considerations: calculate the animal consumption according to body surface area: mice 20g=adult measures 3 * 0.0026=0.0078g; 0.0078 * 50=0.39g/kg (being equivalent to crude drug amount 3.51g/kg) is an equivalent; Middle dosage is 0.39 * 2=0.78g/kg (being equivalent to crude drug amount 7.02g/kg), high dose 0.39 * 3=1.17g/kg (being equivalent to crude drug amount 10.53g/kg).Rat and Cavia porcellus are adult's amount * 0.018 and are the consumption of 200g; As equivalent is 3 * 0.018=0.054g; 0.054 * 5=0.27g/kg (being equivalent to crude drug amount 2.43g/kg); Middle dosage 0.27 * 2=0.54g/kg (being equivalent to crude drug amount 4.86g/kg), high dose 0.27 * 4=1.08g/kg (being equivalent to crude drug amount 9.72g/kg).Dosage all holds with drop pill separates matter quantity representation.

Positive control drug: codeine phosphate, 15mg/ sheet, people 15-30mg every day, Shenyang No. 1 Pharmaceutical Factory, lot number 980907; Ammonium chloride, people 0.5-1g every day, Beijing Chemical Plant's product, analytical pure, lot number 911103; Aminophylline 0.1g/ sheet, people 0.2-1g every day, Shandong XinHua Pharmacy stock Co., Ltd, lot number 20001221; Dexamethasone, Yongning, Jinan pharmaceutical Co. Ltd, 0.75mg/ sheet, people 1.5-7.5mg every day lot number 0011109; Ciprofloxacin, Jinan east wind pharmaceutical factory, 200mg/ sheet, people 200-600mg every day lot number 0108034.

Experimental animal: Kunming mouse, 18-22g, ♀ ♂ half and half, Shandong University's Experimental Animal Center provides, quality certification Shandong kinoplaszm word 20001002; Cavia porcellus is planted by Britain, 180-240g, ♀ ♂ half and half, and Shandong University's Experimental Animal Center provides, the quality certification number: Shandong kinoplaszm word: 20001004; Wister kind rat, Shandong University's Experimental Animal Center provides, quality certification Shandong kinoplaszm word 20001003.

Main agents: phenol red, Shanghai reagent three factories, lot number 920924; Citric acid, 99.5%, Qingdao chemical reagent factory, lot number 930725; Pragmoline (Specia), Shanghai Vaccine and Serum Institute, purity 98%, lot number 940311; Histamine phosphate, Shanghai reagent three factories, purity 98%, lot number 980409.

Key instrument: unico 2100 ultraviolet-visible spectrophotometers, Shanghai close sharp analytical tool company.ZS-200A type electricity separation, ultrasound atomizer, the Central Asia, Tianjin scientific and technological development company.9B303 type electronic balance, prunus mume (sieb.) sieb.et zucc. Teller-Chinese company (Shanghai) product.

Experimental technique and result

The test of 1 antitussive

1.1 ammonia is drawn the antitussive action of coughing mice ^[2]: 60 of 18-20g mices, each 30 of male and female are divided into 5 groups at random; Relieving asthma every group each 12 of drop pill (appearance is separated thing) of normal saline matched group, the celestial spirit of 0.39g/kg, 0.78g/kg and 1.17g/kg, other establishes codeine (Shenyang No. 1 Pharmaceutical Factory product; Lot number 980907; Be made into the suspension of 2mg/ml) 12 of 20mg/kg matched groups, the administration capacity all is 0.2ml/10g, the celestial spirit drop pill liquor strength of relievining asthma is 1.95%, 3.9%, 5.85% suspension.Except that the codeine group, other four groups were successive administration two days, once a day ig; Fasting 10h, 90min behind the medicine, and codeine is fasting 10h; Behind the disposable administration 90min, 5 groups of mices are all put in the 3L beaker successively, are given 25% ammonia atomization 3S by the bottom surface by ZS2001A type ultrasound atomizer; Observe each mouse cough and cough number of times in incubation period and the 2min occurs, t inspection statistics comparable group differences, the result is as shown in table 1.

The celestial spirit of table 1 antitussive action

of drop pill of relievining asthma to ammonia induced mice cough

With normal saline matched group ratio, * * P＜0.01 * * * P＜0.001

Experimental result shows that celestial spirit is relievingd asthma, and cough has tangible antitussive action to drop pill to the ammonia induced mice, not only prolongs the cough response latency, also reduces the cough number of times.

1.2 antitussive action: 180-240g Cavia porcellus, male and female half and half, each 5 of every group of male and female to guinea pig cough due to the citric acid; Be divided into and be physiology saline control group, the celestial spirit drop pill of relievining asthma, 0.27g/kg group (2.7%1ml/100g), 0.54g/kg (5.4%1ml/100g) and 1.08g/kg organize (10.8%1ml/100g); Other establishes codeine 20mg/kg group, and preceding 4 groups are the successive administration secondary, once a day; Fasting 10h, last administration 90min, the codeine group is fasting 10h; Behind the disposable administration 90min, each Cavia porcellus is put in the 10L bell glass, is given 17.5% citron acid solution 1min by the bottom ultrasonic atomizatio; Write down each guinea pig cough and react the time (incubation period) of appearance and the cough number of times in the 5min, t check comparable group differences, the result is as shown in table 2.

The celestial spirit of table 2 antitussive action of drop pill of relievining asthma to guinea pig cough due to the citric acid

With normal saline matched group ratio, * * P＜0.01 * * * P＜0.001

Experimental result shows, the celestial spirit drop pill of relievining asthma has tangible antitussive effect to guinea pig cough due to the citric acid, not only prolongs the incubation period that cough occurs, and reduces the cough number of times in the certain hour.

2 expectorant tests---the phenol red method of mice

60 of 18-22g mices are divided into 5 groups at random, each 6 of every group of male and female, and the celestial spirit drop pill group of relievining asthma is identical with 1.1 with 3 dosage for the normal saline matched group; Other establishes one 2% ammonium chloride group (ammonium chloride, Beijing Chemical Plant's product, analytical pure); The administration capacity is identical with 1.1, successive administration two days, once a day.Fasting 10h, mice gives 0.25% phenol red liquid 0.2ml/10gip (being equivalent to phenol red 50mg/kg) behind the last administration 60min, injects phenol red back 30min, takes off neck and puts to death mice, by 5%NaHCO ₃Each mouse breathing road of liquid 5ml flushing three times (No. 7 syringe needle is peeled off trachea by the Adam's apple place, cuts off, and syringe needle inserts about 0.6cm in the trachea); It is centrifugal to collect flushing liquor 3000r/min * 10min; By 722 grating spectrophotometers (Shanghai analytical tool three factory's products), survey the OD value in the 456nm place, and join 0.01% titer (1ug/ml) survey OD value secondary; Average and be standard value; Each sample OD value/standard value * 1ug/ml is every Mus drug group-matched group of phenol red output/matched group * 100% and is recruitment (%), result such as table 3.

The influence (X ± S ug/ml) of table 3 pair mouse breathing road phenol red output

Experimental result points out the celestial spirit drop pill of relievining asthma obviously to increase mouse breathing road phenol red output, explains of the present invention group

The compound preparation has tangible phlegm-dispelling functions.

3 test-histamine the Cavia porcelluss of relievining asthma are drawn the method for breathing heavily

50 of 180-240g Cavia porcelluss are divided into 5 groups at random, 10 every group; The celestial spirit of normal saline matched group and three dosage is relievingd asthma the drop pill group with 1.2 identical, and positive controls is aminophylline (pharmacy limited company of Shandong Xinhua, lot number a 00001221) 40mg/kg (0.1g/ sheet; 2,0.2ml is made into 50ml 1ml/100gig) the celestial spirit of normal saline group and three dosage relievings asthma drop pill administering mode and dosage with 1.2, successive administration two days; Fasting 10h behind the last administration 90min, puts Cavia porcellus in the 10L bell glass; The equivalent mixed liquor that is given 4% Pragmoline (Specia) (Shanghai Vaccine and Serum Institute provides, purity 98%), 0.2% histamine phosphate (Shanghai reagent three factory's products, purity 98%) liquid by the bottom surface ultrasonic atomizatio (contains acetylcholine 2%; Histamine 0.1%) 15S writes down each Cavia porcellus and pants and incubation period occurs, twitches incubation period and the interior tic number of times of 10min to occur; T check comparable group differences is like table 4.

The celestial spirit of table 4 inhibitory action

of drop pill of relievining asthma to Cavia porcellus asthma due to the histamine

With normal saline matched group ratio, * * P＜0.01 * * * P＜0.001

Experimental result shows, the celestial spirit Cavia porcellus asthma attack that drop pill causes histamine when＞0.7g/kg dosage of relievining asthma has the obvious suppression effect, shows tangible antiasthmatic effect.

The test of 4 antiinflammatories

4.1 inhibition to mice ear:

60 of male 20-22g mices, random packet be with 1.1,12 every group; Positive controls is dexamethasone (Yongning, Jinan pharmaceutical Co. Ltd product, a lot number 001109), and normal saline matched group and celestial spirit are relievingd asthma three dose groups administrations of drop pill with 1.1; Successive administration two days, last administration 30min, positive control drug (dexamethasone 3mg/kg) is disposable administration; Administration 30min, each is organized the equal auris dextra exterior feature of mice and gives 2% compound recipe Oleum Tiglii 20 μ l and be coated with auricle two sides anyway, behind the 4h; Put to death mice, lay left and right sides auricle by the card punch of diameter 9mm, with prunus mume (sieb.) sieb.et zucc. Teller-Tuo benefit 9B303 scales/electronic balance weighing; Auris dextra weight-left ear weight=swelling degree, matched group swelling degree-drug group swelling degree/matched group swelling degree is that medicine presses down scorching rate, result such as table 5.

Table 5 antiinflammatory Asthma capsule causes the influence

of ear swelling to mouse knoting oil

With normal saline matched group ratio, * * P＜0.01 * * * P＜0.001

Experimental result points out the celestial spirit drop pill of relievining asthma that the effect of certain inhibition acute exudative inflammation is arranged, but not obvious, presses down scorching rate＜30%.

4.2 influence to the SN-263 rat paw edema

50 of 180-220 male rats; Be divided into 10 of normal saline matched groups at random, the relieving asthma suspension 1ml/100gig of drop pill 0.27g/kg, 0.54g/kg, three various dose of 1.08g/kg (giving 2.7%, 5.4%, 10.8% respectively) of celestial spirit establishes dexamethasone 1mg/kg positive controls in addition.Three various dose groups of normal saline matched group and eliminating phlegm to relieve asthma were successive administration two days; Behind the fasting 10h, survey earlier before the last administration and do not cause the right sufficient sole of the foot volume of scorching each rat, administration then; And only give 1% carrageenin (institute of Chinese materia medica, Liaoning product) 0.2ml/, left sufficient sole of the foot fascia injected.Dexamethasone is a single administration; Survey normal foot sole of the foot volume before the administration earlier, cause inflammation to SN-263 again, survey right sufficient sole of the foot volume of each rat (be by the graduated special measuring rat foot of threeway pipe racks volume device) respectively respectively at causing scorching back 1,2,4,6,8h then; The variant time is respectively organized the rat paw change in volume; Swelling degree=(causing the scorching back of scorching front volume-cause volume)/cause scorching front volume * 100%, each organizes swelling degree such as table 6, and a statistical is organized in the t check.

The influence of the table 6 pair rat paw edema (ml of X ± S)

With normal saline matched group ratio, * P＜0.01 * * P＜0.01 * * * P＜0.001

Experimental result: the celestial spirit foot swelling that drop pill causes rat carrageenan of relievining asthma has certain inhibitory action, explains that present composition preparation has certain antiinflammatory action.

5. to the bacteriostasis of standard respiratory tract common pathogen

5.1 extracorporeal bacteria inhibitor test---test tube doubling dilution; Introduce staphylococcus aureus ATCC25925 by Shandong Prov. Sanitation and Antiepidemic Station strain chamber; Diplococcus pneumoniae ATCC25273; Beta hemolysis type streptococcus ATCC27027 and escherichia coli ATCC21922 strain, and by strain chamber assistance carrying out extracorporeal bacteria inhibitor test.Culture medium is: soup in the beef nutrition+sheep blood (95: 5), culture medium and the celestial spirit drop pill of relievining asthma holds and separates thing, behind the autoclave sterilization; With the culture medium doubling dilution; Appearance is separated thing be diluted to the 0.00078125g/ml variable concentrations by 0.05g/ml, other establishes and does not contain the antibacterial matched group that the drop pill appearance is separated thing, and each concentration is 10 test tubes; Each test tube all is added on 0.1ml, is diluted to 10 ^-3Fresh bacterium liquid, in 37 ℃ of incubators, behind the 24h, get each test tube microscopy; Observe fungistatic effect, aseptic life (-) has bacteria growing (+); 10 test tubes of every kind of strain all do not have bacterial growth, are minimum Mlc Mic, and result such as table 7 (only are listed as 0.1g/ml～0.00125g/ml) in the table.

The celestial spirit of the table 7 external Mlc Mic of drop pill that relievings asthma

Annotate: (-) is asepsis growth, and (+) is aseptic life ,+for the 1-2 pipe has bacteria growing 10～20%, ++ be 30～40%, +++be 50～80%, ++ ++ be 80～100%, 10 test tubes of culture medium contrast all have bacterial growth 100%.Is 0.00625g/ml by the visible present composition preparation of result of the test to the antibacterial Mic of respiratory tract common pathogen gold Portugal bacterium; Mlc Mic to beta hemolytic streptococcus and Diplococcus pneumoniae is 0.0125g/ml, explains that present composition preparation has certain bacteriostasis.Each strain variable concentrations bacterial growth situation such as table 8 (asepsis growth percentage ratio) %.

Each strain variable concentrations bacterial growth situation of table 8

The asepsis growth test tube is counted ÷ 10 * 100%, is table percent.

5.2 the golden yellow Fructus Vitis viniferae feel of the ball is dyed the protective effect of mice

By economizing epidemic prevention station strain chamber the staphylococcus aureus culture fluid is provided, the gastric Mucin normal saline doubling dilution with 5% is a bacteria containing amount about 9 * 10 ⁸, 4.5 * 10 ⁸, 9 * 10 ⁷, 4.5 * 10 ⁷, 9 * 10 ⁶Five concentration give 18-22g ip in mice, and 10 mices of each concentration were observed 3 days, and record dead mouse situation confirms that minimum whole lethasl concentration is 9 * 10 ⁷

60 of 18-20g mices, each 30 of male and female are divided into 5 groups at random; Normal saline matched group, the celestial spirit drop pill 0.39g/kg that relievings asthma, 0.78g/kg, 1.17g/kg (giving 1.95%, 3.9%, 5.85% suspension respectively); The administration capacity is 0.2ml/10g (with 1.1) all; Positive control drug is ciprofloxacin 0.2g/kg, and successive administration 3 days gives 5% gastric Mucin and is diluted to 9 * 10 after the last administration ⁷Fresh staphylococcus aureus ATCC25925 bacterium liquid 0.2ml/10g ip, carry out germ attack.Record: respectively organize the dead mouse situation in 7 days, after the tapping 24 hours, 48 hours, each organized administration respectively.Experimental result such as table 9.

The protective effect of table 9 pair infection of staphylococcus aureus mice

Experimental data is pressed qualitative response statistics method-X by medical experiment statistical software of Huaxi Medical Univ ²Check, exact propability computer statistics such as following table 10.

Table 10

Presentation of results, the celestial spirit drop pill of relievining asthma has the certain protection effect to the infection of staphylococcus aureus mice, reduces mortality rate.

6. to the bacteriostasis of the clinical separation common pathogen of respiratory tract

6.1 external bacteriostasis, this experiment is assisted to accomplish result such as table 11 by national related request by epidemic prevention station strain chamber, Shandong Province

The celestial spirit of table 11 is relievingd asthma drop pill to bacteriostasis in the body of clinical separation respiratory tract common pathogen

Annotate: "+" no bacterial growth promptly has inhibitory action; "-" has bacterial growth is the unrestraint effect

The celestial spirit drop pill of relievining asthma is respectively 3.12-25,3.12-25,3.12-12.5,1.56-12.5mg/ml to the MIC of staphylococcus aureus, beta hemolysis type streptococcus, streptococcus pneumoniae and hemophilus influenza.The celestial spirit drop pill of relievining asthma is all inhibited to the main pathogenic bacterium staphylococcus aureus, beta hemolysis type streptococcus, streptococcus pneumoniae, the hemophilus influenza that cause upper respiratory tract infection.

6.2 protective effect to clinical separation pneumococcal infection mice

According to the trial test result, 110 of 18-22g mices, male and female half and half are divided into 5 groups at random, each 11 of every group of ♀ ♂, the grouping administration is with 5.2, successive administration three days, giving by the bacteria containing amount that epidemic prevention station strain chamber, Shandong Province provides behind last administration 2h, the fasting 12h is 8 * 10 ⁷Individual/the ml streptococcus pneumoniae (economize epidemic prevention station and be located away from province Qianfo Mount hospital Pneumology Department, strain number 98065 in 98 years) culture fluid increases venom 0.2ml/10g, ip by the stomach film; Observe 72h; The dead mouse number respectively organized in record, calculates mortality rate, by exact propability check carrying out statistical result such as table 12

The celestial spirit protective effect of drop pill of relievining asthma to the pneumococcal infection mice

The mice major part is dead in vaccine is attacked 24h, and dead mice is the blood stasis shape. and each administration group and water matched group carry out statistical test by exact propability, result's celestial being spirit drop pill 0.39g/kg that relievings asthma, P＜0.05; 0.78g/kg group and 1.17g/kg group, P all＜0.01; Ciprofloxacin 0.2g/kg organizes P＜0.01.

Explain that present composition preparation has the significant protection effect to respiratory tract common pathogen pneumococcal infection mice.

Conclusion (of pressure testing):

Observing present composition preparation by effect experiment all has the obvious suppression effect to guinea pig cough due to ammonia induced mice cough, the citric acid, shows tangible antitussive action; Obviously increase mouse breathing road phenol red output, show tangible phlegm-dispelling functions, Cavia porcellus asthma due to the histamine is had the significant protection effect, show significant antiasthmatic effect; In addition rat and mice exudative inflammation all there is certain inhibitory action; Experiment in vitro confirms that present composition preparation has certain bacteriostasis to the respiratory tract common pathogen, has the significant protection effect to staphylococcus aureus and pneumococcal infection mice.Pharmacodynamic experiment show present composition preparation have sure relieving asthma, eliminate the phlegm, cough-relieving and certain antiinflammatory action, the respiratory tract common pathogen is also had certain bacteriostasis, be applicable to the traditional Chinese medical science " bronchial wheezing " patient of lung kidney deficiency.

Experimental example 2 clinical experiments

[kind of design and principle]

One, research design type: guiding clinical trial; Adopt the research design of segmentation random packet, contrast (3: 1), double blinding.

Two, the sample size of clinical experimental study needs: sample size is: 40 examples.

Three, segmentation random method: adopt SASV8 software, carry out the segmentation random packet; 40 examples are in hospital or the out-patient, are divided into treatment group and matched group, and 30 examples are organized in treatment, matched group 10 examples.

Four, blind method design

Adopt the double blinding measuring method; Execute blind to researcher (gauger is medical and technical staff, data management person and statistical analysis worker) and object of study (patient).

Five, contrast

Test group: the doctor trained in Western medicine conventional therapy+celestial spirit side of relievining asthma medicine

Matched group: doctor trained in Western medicine conventional therapy+classical side's medicine jade screen looses and closes seven flavor Duqi Wans.

Explain:

Same standard is adopted in the routine of western medicine treatment, Slow-release Theopylline, and 0.1 gram/every, each a slice, per morning and evening is respectively once.

The celestial spirit side of relievining asthma medicine: Herba Epimedii 9g Folium Elaeagni 12g, Herba Epimedii 9g, Fructus Schisandrae Chinensis 6g, decoction.

Classics side's medicine jade screen looses and closes seven flavor Duqi Wans: Radix Astragali Preparata 15 grams, windproof 10 grams, the Rhizoma Atractylodis Macrocephalae 10 grams, Radix Rehmanniae Preparata 12 grams, Cortex Moutan 10 grams, Rhizoma Alismatis 10 grams, Poria 12 grams, Fructus Corni 10 grams, Rhizoma Dioscoreae 12 grams, Fructus Schisandrae Chinensis 6 grams, decoction

[study population]

One, study population's physical data

(1), sex, age

Treatment group and matched group men and women constituent ratio, P＞0.05, no significant difference has comparability.

Treatment group and matched group patient age compare, and P＞0.05 has comparability.

(2), occupation distributes: the treatment group and the matched group course of disease compare, and P＞0.05 has comparability.

(3) course of disease

Table 13 course of disease relatively

The treatment group and the matched group course of disease compare, and P＞0.05 has comparability.

(4) cardinal symptom, sign

Table 14 symptom and sign relatively

Treatment group and matched group main disease, levy aspect relatively, P＞0.05 has comparability.

To sum up data shows, treatment group and matched group patient's basic document is similar, has comparability.

Two, the study population's confirms

(1) medical diagnosis on disease, state of an illness classification and curative effect determinate standard

1 san bronchial asthma diagnostic criteria: Chinese Medical Association's respiratory disease credit meeting asthma group " prevention and control of bronchial asthma guide (2003) ".The chronic duration was meant in considerable time, equal weekly different frequency with (or) symptom (pant, out of breath, uncomfortable in chest, cough etc.) appears to some extent.

The division of 2 feelings degree:

2.1 pant

Slightly (+): the idol of panting has outbreak, and degree is light, does not influence sleep or movable.

Moderate (++): the state of an illness is between slight (+) and severe (+++).

Severe (+++): pant obviously, can not put down for sleeping in, influence sleep and movable.

2.2 cough

Slightly (+): be interrupted cough daytime, do not influence orthobiosis and work.

Moderate (++): symptom is between slight (+) and severe (+++).

Severe (+++): the frequent or apasm of coughing of cough round the clock, influence is had a rest and sleep.

2.3 cough up phlegm

Few (+): the 10～15ml that coughs up phlegm round the clock, or the 5～25ml that coughs up phlegm in night and early morning.

In (++): the 51～100ml that coughs up phlegm round the clock, or the 26～50ml that coughs up phlegm in night and early morning.

Many (+++): cough up phlegm more than the 100ml round the clock, or night and early morning cough up phlegm more than the 50ml.

Sputum characteristic, color, observed and recorded in addition.

2.4 wheezing sound

Few (+): idol is heard, or occurs in cough, the back of deeply breathing.

In (++): be dispersed in.

Many (+++): be abound with.

The judgement of clinical severity extent, symptom and pulmonary's wheezing sound during by prescription on individual diagnosis, any one enough severe person is a severe, and enough moderate persons are moderate, and equal not enough person is slight.

The syndrome differential diagnosis standard of the traditional Chinese medical science lung kidney deficiency of chronic duration of 3 bronchial asthmas is (with reference to State Standard of the People's Republic of China GB/16751.2-1997; " tcm clinical practice diagnosis and treatment term syndrome part. lung system card class "; The 1997.3.4 of State Bureau of Technical Supervision issue, 1997.10.01 implements to formulate)

Pant, out of breath, uncomfortable in chest, cough, moving then for very; Or it is unable to cough; Or it is rare clearly to spit, or it is many to spit; Low voice, or spontaneous perspiration is arranged, anemophobia; Light red tongue is fat, and tongue is greasy in vain, deep-thready pulse or a little less than.

(2) test case standard:

1 includes the case standard in:

1.1 the chronic duration of bronchial asthma, (symptom＜1 time weekly of short durationly SOA≤every month 2 times at night occurred, and F EV1 accounts for predicted value %>=80% or P E F>=80% people's optimum, P E F or F E V1 aberration rate＜20%; Or symptom>=1 time weekly, but＜every day 1 time, possiblely influence activity and sleep; SOA＞every month 2 times at night; But＜1 time weekly, F E V 1 accounts for predicted value %>=80% or P E F>=80% people's optimum, P E F or F E V 1 aberration rate 20%～30%) 1.2 age 18-70 year;

1.3 be in the chronic duration of bronchial asthma, nearly apnea road all around infects and acute attack;

1.4 Syndrome in TCM belongs to: lung kidney deficiency person;

[therapeutic scheme]

One, intervenes medicine

Test group: the celestial spirit of the Chinese medicine prescription decoction+routine of western medicine treatment of relievining asthma

Matched group: Chinese medicine YUPINGFENG SAN closes seven flavor Duqi Wan prescription decoction+routine of western medicine treatments.

Two, application method

Chinese crude drug meets the requirements, and adopts decoction method (SOP), the instructions of taking of uniform requirement.

Same standard is adopted in the routine of western medicine treatment, Slow-release Theopylline, and the 0.1g/ sheet, sooner or later respectively once.

Relieving asthma prescription decoction, Chinese medicine YUPINGFENG SAN of the celestial spirit of Chinese medicine closes seven flavor Duqi Wans prescription decoctions and carries out unitizings, makes every effort to the outward appearance unanimity.

[contrast measure]

Test group: the celestial spirit of the routine of western medicine treatment+Chinese medicine prescription decoction of relievining asthma.

Matched group: routine of western medicine treatment+Chinese medicine YUPINGFENG SAN closes seven flavor Duqi Wan prescription decoctions

[curative effect determinate standard]

One, symptom, sign standards of grading

(1), cough

0 minute: do not have cough; 1 minute: be interrupted cough, do not influence orthobiosis and work.

2 minutes: between slight and severe cough.3 minutes: cough round the clock was frequent, influences work and sleep.

(2), cough up phlegm

0 minute: do not have

1 minute: few (+): the 10～50ml that coughs up phlegm round the clock, or the 5～25ml that coughs up phlegm in night and early morning.

2 minutes: in (++): the 51～100ml that coughs up phlegm round the clock, or the 26～50ml that coughs up phlegm in night and early morning.

3 minutes: many (+++): cough up phlegm more than the 100ml round the clock, or night and early morning cough up phlegm more than the 50ml.

Sputum characteristic, color, observed and recorded in addition.

(3), pant

0 minute: do not have

1 minute: slight (+): the idol of panting had outbreak, and degree is light, did not influence sleep or movable.

2 minutes: moderate (+++): the state of an illness was between slight (+) and severe (+++).

3 minutes: severe (+++): pant obviously, can not put down for sleeping in, influence sleep and movable.

(4), wheezing sound

0 minute: do not have; 1 minute: few (+): idol was heard, or occurred in cough, the back of deeply breathing.

2 minutes: in (++): be dispersed in; 3 minutes: many (+++): be abound with.

Two, curative effect judging standard

(1), individual event doing well,improving situation

1 clinic control: transference cure.2 produce effects: symptom changed into 1 fen by 3 minutes.

3 take a turn for the better: symptom changed into 2 fens by 3 minutes or changed into 1 fen by 2 minutes.4 is invalid: symptom does not have improvement or increases the weight of.

(2), the judgement of comprehensive clinical efficacy

According to coughing, cough up phlegm, panting, the situation of change of total score of wheezing sound clinical symptoms sign scoring is judged.Total score * 100% before improvement rate (%)=(the total score in total score-treatment back before the treatment) ÷ treatment

1 clinic control: improvement rate >=85%; 2 produce effects: improvement rate 60%～84%; 3 take a turn for the better: improvement rate 30%-59%; 4 is invalid: improvement rate＜30%.

[result and analysis]

One, clinical condition is levied interpretation of result

(1), individual event symptom, sign curative effect

Table 15 individual event symptom, sign curative effect are relatively

Table 15 prompting treatment group and matched group comparison have similarity aspect cough-relieving, through statistical procedures (Ridit analysis) the no significant difference in P＞0.05; Relievining asthma and improving that the treatment group is superior to matched group aspect the wheezing sound sign, and apparent control rate is higher than matched group, P＜0.05 significant difference.

(2), multinomial symptom, sign (cough, expectorant, breathe heavily, wheezing sound) comprehensive therapeutic effect relatively

The multinomial symptom of table 16, sign comprehensive therapeutic effect are relatively

Table 16 pair treatment group and the multinomial symptom of matched group, sign (cough, expectorant, breathe heavily, wheezing sound) comprehensive therapeutic effect compares, results suggest treatment group is better than matched group.Through statistical procedures P＜0.05 significant difference.

Two, laboratory result analysis

(1), lymphocyte transformation experiment and complement C3 change relatively before and after treatment group and the treatment of control group

Lymphocyte transformation experiment and complement C3 change relatively

before and after table 17 treatment group and the treatment of control group

Table 17 prompting: two groups of patients are after treatment, and lymphocyte transformation experiment and complement C3 content all have increasing in various degree, handle P＜0.05 through statistics (t check), present remarkable meaning, and explaining to treat for two groups all has immune defence potentiation.Compare zero difference (P＞0.05) between two groups, explain that tentatively the two has certain concordance.

(2), PGE before and after treatment group and the treatment of control group ₂, TXB ₂Change relatively

PGE2, TXB2 change relatively

before and after table 18 treatment group and the treatment of control group

Table 18 treatment group treatment back PGE ₂Content increases, and there were significant differences to handle P＜0.05 through statistics (t check); Though matched group has increase, P＞0.05 not statistically significant.This preliminary explanation treatment group is being alleviated bronchospasm, and the reduction airway resistance is better than matched group with the effect of the aspect of relievining asthma.After two groups of treatments, TXB ₂Content all has decline in various degree, P＜0.05 significant difference, P between group＞0.05 zero difference.Tentatively show two groups reducing pulmonary vascular resistance, regulate the lung microcirculation, further improving the pulmonary function aspect has concordance.

(3), IgG IgA IgM (enzyme linked immunosorbent assay mensuration) relatively before and after treatment group and the treatment of control group

IgG IgA IgM (enzyme linked immunosorbent assay mensuration) compares

before and after table 19 treatment group and the treatment of control group

Table 19 prompting: two groups of treatments all influence content of immunoglobulin IgG to some extent; Be after the treatment in various degree and reduce; Self compare P＜0.05 before and after two groups of treatments, there were significant differences on the statistics, explains that tentatively two groups of correlative factors to air flue spasm and obstruction have the positivity influence.

(4), blood IgE IgA before and after treatment group and the treatment of control group; Blood IgE IgA before and after expectorant IgE IgA (putting the method for exempting from measures) comparison sheet 20 treatment groups and the treatment of control group, expectorant IgE IgA (putting the method for exempting from measures) relatively

After two groups of treatments of table 20 prompting, expectorant, blood IgA content improve, and expectorant, blood IgE content descend, through statistical procedures P＜0.05, and P between group＞0.05.Show that tentatively two groups have positive role aspect the adjusting of the immune defence mechanism of lung.

(5), pulmonary function changes relatively before and after treatment group and the treatment of control group

Pulmonary function changes relatively

before and after table 21 treatment group and the treatment of control group

Table 21 prompting treatment group and matched group all have improvement in various degree to pulmonary function, and the treatment group is more obvious.Through between two groups of treatment front and back of statistical procedures self and group, relatively all significant difference is arranged P＜0.05 through statistical procedures.

Experimental example 3 preparation researches

1, choice of base

In the present composition active ingredient be mostly that molecular weight is big, slightly solubility material, thermal instability composition, in order to improve the medicine stripping, and at utmost improve the drug loading of medicine, very important for choice of base.

The substrate of drop pill can be divided into the fat-soluble fat-soluble substrate of using always with water solublity two big classes has stearic acid, glyceryl monostearate, insect wax, Cera Flava, paraffin, hydrogenated vegetable oil, octadecanol etc., and water-soluble base commonly used has polyethylene glycols, sodium stearate, carbamide polyethers poloxamer etc.

Show that through trial test when making substrate with PEG4000, the hardness of drop pill, roundness and thermostability are all relatively poor.

This test is an index with roundness, the hardness of drop pill, and the mixture of investigating both different proportions is a substrate.Roundness refers to the ratio of the minor axis of drop pill and major diameter, and hardness obtains the result of hardness through the method for finger pressure, and the result is divided into four grades (general, better, good, fine) result of the test and sees the following form

Table 22 substrate determination of ratio (n=20)

Experimental result shows PEG4000: the variation of PEG6000 ratio is not obvious to drop pill hardness, but influential to roundness, confirms PEG4000 according to each item index: the PEG6000 ratio is 8: 1.

2. drug loading confirms

2.1 the initial option of drug loading

Select for use Polyethylene Glycol (PEG4000: PEG6000 is 8: 1) for substrate, particle diameter, roundness (the minor axis of drop pill/major diameter), the ball method of double differences of investigating drug loading and be 15%, 20%, 25%, 30% o'clock drop pill are different.The result sees the following form.

Definite (n=20) of table 23 drug loading

The result shows that drug loading is poor 35% o'clock smelt flow property, and not easy-formation, and particle diameter, roundness, the ball method of double differences are different big, and be also bigger 15% o'clock each item index difference.Therefore in 20%, 25%, 30%, select ratio of greater inequality.

2.2 drip the optimization of system condition

2.2.1 the investigation of melt temperature

By above definite prescription, substrate to be put in the water-bath of different temperatures and heat, fusion adds medicine and stirs, and drips system.

With the whole degree of pill shaped circle, oozing difficulty or ease is index, and the result sees the following form.

Table 24 melt temperature is investigated (n=20)

The result shows that melt temperature is good 75 ℃ of roundness, and is prone to ooze.Its main cause is that temperature raises, and Polyethylene Glycol viscosity reduces, and mobile the enhancing helps medicine and substrate mix homogeneously; Be unfavorable for molding but temperature is too high.So selected melt temperature is 75 ℃.

2.2.2 the investigation of condensing agent temperature

By above definite method, substrate to be put in 75 ℃ the water-bath and heat, fusion adds medicine, stirs, and system is dripped in 75 ℃ of insulations, splashes in the coolant of different temperatures, be that the index result sees the following form with the whole degree of pill shaped circle.

Table 25 condensing agent temperature is investigated (n=20)

The result shows that the drop pill roundness was good, and difference is little when condensation temperature was controlled at 2 ℃-6 ℃.Its main cause is that temperature reduces, and condensing agent viscosity increases, and the drop pill translational speed reduces thereupon, has reduced the influence of translational speed, helps improving the drop pill roundness.But condensate temperature is low excessively, can make drop pill solidify quickening, is unfavorable for that drop pill shrinks and the eliminating bubble, reduces roundness on the contrary.This test is immersed in the dark ice-water bath of 10cm the condensed fluid hypomere, has formed thermograde, and gained drop pill roundness is good, therefore confirms the condensed fluid lower part is immersed in 2 ℃-6 ℃ the ice-water bath.

2.2.3 the optimization of drip the system temperature, dripping distance, cooling off conditions such as column length

An important indicator of drop pill quality evaluation is the different measurement of the ball method of double differences, and the ball method of double differences is different more little, and content distribution is even more, and dosage is more accurate.It is many to influence the heavy difference factor of ball, but mainly contains fluid temperature, a distance, cooling column length.Different with the ball method of double differences is index, and fluid temperature (A), a distance (B), cooling column length (C) are designed quadrature L9 (34) by table 1-33 and show, and does 3 factors, 3 hydraulic tests screening.

Table 26 orthogonal design factor level table

Different with the ball method of double differences is index, and result of the test and preferred technology are following:

Table 27 droplet system condition orthogonal test and result

The different finger of the ball method of double differences is got test sample 20 balls, accurate fixed every ball weight, the heavy RSD value of calculating ball claimed.

Table 28 a system condition orthogonal test variance analysis

Annotate: the P value is a significant difference less than 0.05, is that difference is extremely remarkable less than 0.01.

The results of analysis of variance shows fluid temperature (A), drip apart from there are no significant in the EXPERIMENTAL DESIGN scope influence of (B), cooling column length (C).The optimised process of intuitive analysis is A ₁B ₂C ₃, promptly drip 70-75 ℃ of system temperature, drip apart from 4cm cooling column length 75cm.

2.2.4 demonstration test

Take by weighing 2 parts of raw material and adjuvants in the prescription ratio, according to preferred technology A ₁B ₂C ₃(promptly drip 70-75 ℃ of system temperature, drip apart from 4cm, cooling column length 75cm) experimentizes, with optimum experiment A in the orthogonal table ₃B ₁C ₃Compare.

Table 29

Result: press A ₁B ₂C ₃Carry out twice test, the different index of the ball method of double differences all is superior to optimum experiment A in the orthogonal table ₃B ₁C ₃(1.92).So confirm that technology is A ₂B ₂C ₂, promptly drip 70-75 ℃ of system temperature, drip apart from 4cm cooling column length 75cm.

2.3 process certification test

By the prescription of confirming and drip 3 batches of drop pill of system condition preparation, it is different to measure particle diameter, roundness, the ball method of double differences respectively, and the result sees the following form 30.

Table 30 3 lot sample article process certifications test

It is feasible that the result shows that prescription reaches a system process stabilizing.

Following embodiment all can realize the effect of above-mentioned experimental example.

Embodiment 1:

Folium Elaeagni 12g, Herba Epimedii 9g, Fructus Schisandrae Chinensis 6g;

The Folium Elaeagni water boiling and extraction, each 10 times of amounts, 1 hour are extracted 3 times; Merge extractive liquid, filters, and filtrating is concentrated into 1: 1, and (medical material: filtrating is g: ml); Adopt the macroporous resin enrichment effective ingredient, the sample solution concentration of extract is 0.1g crude drug/ml, appearance on the speed of 2BV/h; The water elution remove impurity of 4BV/h uses 60% ethanol to be the eluant eluting, and elution flow rate is that 3BV/h, elution volume are 4BV; Behind the eluent decompression recycling ethanol, drying under reduced pressure is pulverized to fine powder subsequent usely, is Folium Elaeagni extract I;

After Fructus Schisandrae Chinensis is broken for coarse granule, 8 times of amount 50% ethanol, extracted 1.5 hours, 1.5 hours respectively, merge extractive liquid, filters, and decompression filtrate recycling ethanol is concentrated into relative density and is 1.15-1.25 (50 ℃), and is subsequent use;

Folium Epimedii adds 30 times of water yields of water at every turn, decocts to extract each 1.5 hours 3 times; Extracting solution filters, and merging filtrate is condensed into 1: 1 (medical material g: filtrating ml); Select the rich composition of imitating of macroporous resin for use, sample solution concentration is 0.5g crude drug/ml; Applied sample amount is 1g crude drug/ml resin; Adsorption rate is 0.5ml/min; Elution speed is 1.0ml/min; The water elution volume is 10BV; Eluting solvent is 70% ethanol, and consumption is 6BV; Behind the eluent decompression recycling ethanol, drying under reduced pressure, pulverize into fine powder subsequent use;

Above-mentioned Folium Elaeagni extract I mixes with Fructus Schisandrae Chinensis extrat, Herba Epimedii extract, and the drug loading adding substrate with 30% (PEG4000: PEG6000=8: 1), mixing, condensation temperature is controlled at 2 ℃-6 ℃; Dripping the system condition is: drip 70-75 ℃ of system temperature, drip apart from 4cm, cooling column length 75cm; The preparation drop pill.

Dosage (dosage on the one): the 27g/ crude drug/everyone/day

Function: antiasthmatic-antitussive, element culturing fixed folder.

Main syndrome (indication): be mainly used in the lung kidney deficiency the traditional Chinese medical science " bronchial wheezing " (clinically mainly show as: pant, out of breath, uncomfortable in chest, cough, moving then for very; ); The bronchial asthma (Bronchial athma) that modern medicine is alleged wherein especially with the chronic duration of bronchial asthma, belongs to this category, can be by this disease Coryza Treated by Syndrome Differentiation.(the chronic duration is meant in considerable time, equal weekly different frequency and (or) occur to some extent symptom (pant, out of breath, uncomfortable in chest, cough etc.)); The consumption usage: each 20, every day 3 times, buccal or warm water delivery service after meal.

The course of treatment: two weeks (14 days) are a course of treatment.

Embodiment 2:

Folium Elaeagni 12g, Herba Epimedii 9g, Fructus Schisandrae Chinensis 6g

Folium Elaeagni, Herba Epimedii, Fructus Schisandrae Chinensis water boiling and extraction, each 20 times of amounts, 1 hour are extracted 3 times, and merge extractive liquid, filters, and filtrating concentrates, and processes soft capsule through conventional technology.

Embodiment 3:

Folium Elaeagni 12g, Herba Epimedii 9g, Fructus Schisandrae Chinensis 6g;

Folium Elaeagni, Herba Epimedii, Fructus Schisandrae Chinensis water boiling and extraction, each 15 times of amounts, 1 hour are extracted 3 times; Merge extractive liquid, filters, the filtrating concentrate drying, and the drug loading with 30% adds substrate (PEG4000: PEG6000=8: 1); Mixing, condensation temperature are controlled at 2 ℃-6 ℃; Dripping the system condition is: drip 70-75 ℃ of system temperature, drip apart from 4cm, cooling column length 75cm;

The preparation drop pill.

Embodiment 4:

Folium Elaeagni 10g, Herba Epimedii 9g, Fructus Schisandrae Chinensis 5g;

Embodiment 5:

Folium Elaeagni 14kg, Herba Epimedii 9kg, Fructus Schisandrae Chinensis 7kg;

Above-mentioned Folium Elaeagni extract I mixes through conventional technology with Fructus Schisandrae Chinensis extrat, Herba Epimedii extract and processes tablet.

Embodiment 6:

Folium Elaeagni 12kg, Herba Epimedii 9kg, Fructus Schisandrae Chinensis 6kg;

60% ethanol extraction of 8 times of amounts extracted respectively 1.5 hours, 1.5 hours, and merge extractive liquid, filters, and decompression filtrate recycling ethanol is concentrated into relative density and is 1.15-1.25 (50 ℃), and is subsequent use, is Folium Elaeagni extract;

Folium Epimedii adds 30 times of water yields of water at every turn, decocts to extract 3 times, and each 1.5 hours, concentrate drying was subsequent use;

Above-mentioned Folium Elaeagni extract mix with Fructus Schisandrae Chinensis extrat, Herba Epimedii extract with 30% drug loading add substrate (PEG4000: PEG6000=8: 1), mixing, condensation temperature is controlled at 2 ℃-6 ℃; Dripping the system condition is: drip 70-75 ℃ of system temperature, drip apart from 4cm, cooling column length 75cm; The preparation drop pill.

Embodiment 7:

Folium Elaeagni 12kg, Herba Epimedii 9kg, Fructus Schisandrae Chinensis 6kg;

Folium Elaeagni 6kg water boiling and extraction, each 10 times of amounts, 1 hour are extracted 3 times; Merge extractive liquid, filters, and filtrating is concentrated into 1: 1, and (medical material: filtrating is g: ml); Adopt the macroporous resin enrichment effective ingredient, the sample solution concentration of extract is 0.1g crude drug/ml, appearance on the speed of 2BV/h; The water elution remove impurity of 4BV/h uses 60% ethanol to be the eluant eluting, and elution flow rate is that 3BV/h, elution volume are 4BV; Behind the eluent decompression recycling ethanol, drying under reduced pressure is pulverized to fine powder subsequent usely, is Folium Elaeagni extract I;

60% ethanol extraction that Folium Elaeagni 6kg8 doubly measures extracted respectively 1.5 hours, 1.5 hours, and merge extractive liquid, filters, and decompression filtrate recycling ethanol is concentrated into relative density and is 1.15-1.25 (50 ℃), and is subsequent use, is Folium Elaeagni extract II.

Above-mentioned Folium Elaeagni extract I and II mix with Fructus Schisandrae Chinensis extrat, Herba Epimedii extract, and the drug loading adding substrate with 30% (PEG4000: PEG6000=8: 1), mixing, condensation temperature is controlled at 2 ℃-6 ℃; Dripping the system condition is: drip 70-75 ℃ of system temperature, drip apart from 4cm, cooling column length 75cm; The preparation drop pill.

Claims

1. Chinese medicine composition of treating bronchial asthma is characterized in that crude drug consists of:

2. Chinese medicine composition as claimed in claim 1 is characterized in that crude drug consists of:

3. Chinese medicine composition as claimed in claim 2 is characterized in that crude drug consists of:

4. like the method for preparing of described any one Chinese medicine composition of claim 1-3, it is characterized in that this method is: Folium Elaeagni, Herba Epimedii, Fructus Schisandrae Chinensis add the pharmacy acceptable auxiliary and process capsule, tablet, granule, gel, slow releasing agent, oral liquid or drop pill after conventional process is extracted; Said common process method comprises water extraction or alcohol extraction or first water extraction water extraction again after alcohol extraction or the first alcohol extraction again.

5. like the method for preparing of described any one Chinese medicine composition of claim 1-3, it is characterized in that this method comprises the steps:

The extraction of steps A, Folium Elaeagni effective ingredient: the Folium Elaeagni water boiling and extraction, each 8-12 doubly measures, 0.5-2 hour, extracts 2-4 time; Merge extractive liquid, filters, and filtrating concentrates; Adopt the macroporous resin enrichment effective ingredient, the water elution remove impurity uses 50-70% ethanol to be the eluant eluting; Behind the eluent decompression recycling ethanol, drying under reduced pressure is pulverized to fine powder subsequent usely, makes Folium Elaeagni extract I; Or the 50-70% ethanol extraction doubly measured with 7-10 of Folium Elaeagni 2-3 time, extracting respectively 1-3 hour, merge extractive liquid, filters, and decompression filtrate recycling ethanol concentrates subsequent usely, makes Folium Elaeagni extract II;

The extraction of step B, Fructus Schisandrae Chinensis effective ingredient: Fructus Schisandrae Chinensis 7-10 doubly measures the 40-60% ethanol extraction 2-3 time, extracts respectively 1-3 hour, and merge extractive liquid, filters, and decompression filtrate recycling ethanol concentrates subsequent use;

The extraction of step C, epimedium active constituent: Folium Epimedii adds the water 20-40 times water yield at every turn, decocts to extract 2-4 time each 1-3 hour; Extracting solution filters, and merging filtrate concentrates; Adopt the macroporous resin enrichment effective ingredient, the water elution volume is 6-12BV; Eluting solvent is a 60-80% ethanol; Behind the eluent decompression recycling ethanol, drying under reduced pressure, pulverize into fine powder subsequent use;

Step D, with above-mentioned Folium Elaeagni extract I or II, or I and II and Fructus Schisandrae Chinensis extrat, Herba Epimedii extract mix homogeneously add the pharmacy acceptable auxiliary and process the acceptable any conventional dosage form of pharmaceutics.

6. the method for preparing of Chinese medicine composition as claimed in claim 5 is characterized in that this method comprises the steps:

Steps A: the Folium Elaeagni water boiling and extraction, each 10 times of amounts, 1 hour are extracted 3 times; Merge extractive liquid, filters, and filtrating is concentrated into 1: 1, medical material: filtrating is g: ml; Adopt the macroporous resin enrichment effective ingredient, the sample solution concentration of extract is 0.1g crude drug/ml, appearance on the speed of 2BV/h; The water elution remove impurity of 4BV/h uses 60% ethanol to be the eluant eluting, and elution flow rate is that 3BV/h, elution volume are 4BV; Behind the eluent decompression recycling ethanol, drying under reduced pressure is pulverized to fine powder subsequent usely, makes Folium Elaeagni extract I; Or Folium Elaeagni extracted respectively 1.5 hours, 1.5 hours with 60% ethanol extraction of 8 times of amounts, and merge extractive liquid, filters, and decompression filtrate recycling ethanol is concentrated into 50 ℃ of relative densities and is 1.15-1.25, and is subsequent use, makes Folium Elaeagni extract II;

After step B, Fructus Schisandrae Chinensis are broken for coarse granule, 8 times of amount 50% ethanol, extracted 1.5 hours, 1.5 hours respectively, merge extractive liquid, filters, and decompression filtrate recycling ethanol is concentrated into 50 ℃ of relative densities and is 1.15-1.25, and is subsequent use;

Step C, Folium Epimedii add 30 times of water yields of water at every turn, decoct to extract each 1.5 hours 3 times; Extracting solution filters, and merging filtrate was condensed into 1: 1, medical material g: filtrating ml; Select the macroporous resin enrichment effective ingredient for use, sample solution concentration is 0.5g crude drug/ml; Applied sample amount is 1g crude drug/ml resin; Adsorption rate is 0.5ml/min; Elution speed is 1.0ml/min; The water elution volume is 10BV; Eluting solvent is 70% ethanol, and consumption is 6BV; Behind the eluent decompression recycling ethanol, drying under reduced pressure, pulverize into fine powder subsequent use.

7. like said any one the drop pill of Chinese medicine composition of claim 1-3, it is characterized in that drop pill substrate percentage by weight wherein is 67%-80%, substrate is PEG4000 and PEG6000, PEG4000: PEG6000=6-11: 1.

8. the method for preparing of drop pill as claimed in claim 7 is characterized in that this method is: the drug loading with 30% adds among PEG4000 and the PEG6000, wherein PEG4000: PEG6000=6-11: 1, and mixing, condensation temperature is controlled at 2 ℃-6 ℃; Dripping the system condition is: drip 70-75 ℃ of system temperature, drip apart from 4cm, cooling column length 75cm.

As described any one Chinese medicine composition of claim 1-3 relieving asthma in preparation, eliminate the phlegm, cough-relieving, antiinflammatory, inhibition staphylococcus aureus or suppress the application in the streptococcus pneumoniae medicine.

10. like the application of described any one Chinese medicine composition of claim 1-3 in preparation treatment chronic bronchial asthma medicine.

11. method as claimed in claim 5 is characterized in that said regular dosage form is capsule, tablet, granule, gel, slow releasing agent, oral liquid or drop pill.