CN101053600A - Medicinal composition for treating cough and preparation method and application thereof - Google Patents

Abstract

The invention relates to a medicine combination for treating cold belonging to medical field. The material of the active components are as followed by weight: kudzu root 4-8, ephedra herb 2-4, cassia twig 2-4, paeonia lactiflora pallas 2-4, bitter almond kernel 2-4. The invention has a plurality of functions such as remarkable bacteriostasis, antivirus, relieving fever, analgesic effect, antiinflammatory, relieving cough, eliminating phlegm and promoting red cell immunity so as to provide a new choose for clinical treatment for cold, especially exogenous pathogenic fever disease.

Description

Pharmaceutical composition of treatment flu and its production and use

Technical field

The present invention relates to a kind of pharmaceutical composition for the treatment of flu, particularly, is to be the medicine that primary raw material is prepared from the natural drug, belongs to field of medicaments.

Background technology

According to theory of Chinese medical science, the etiology and pathogenesis of affection of exogenous wind-cold heating card is analyzed as follows: interior through say " people's wound in cold also, then be febrile disease ", point out that heating is to experience the cold-evil because of body, interspaces of skin and muscles being close, yang-energy must not declare lead to due to.Therefore, on method of treatment, should follow the principle of " high fever like the burning coal over the body, sweating and loose ".

The Chinese patent medicine of state's internal therapy affection of exogenous wind-cold card mainly contains HUOXIANG ZHENGQI WAN (or HUOXIANG ZHENGQI SHUI), Chuan Xiong Tea ball (or Chuan Xiong Tea oral liquid), Jingfang toxicity-removing ball, chill exterior GANMAO CHONGJI etc. at present, but it is mainly as follows that these preparation functions cure mainly, and is mainly used in the vomiting and diarrhoea of treatment affection of exogenous wind-cold internal injury humidity hysteresis as HUOXIANG ZHENGQI WAN (or HUOXIANG ZHENGQI SHUI); Chuan Xiong Tea ball (or Chuan Xiong Tea oral liquid) cures mainly the headache of affection of exogenous wind-cold blockage of main and collateral channels; The Jingfang toxicity-removing ball cures mainly wind and cold visitor table, and the gesture of heat-transformation is arranged then.

Shang Weijian is the relevant report that material combination is used for the treatment of the affection of exogenous wind-cold card with Radix Puerariae, Herba Ephedrae, Ramulus Cinnamomi, the Radix Paeoniae Alba, Semen Armeniacae Amarum, Rhizoma Zingiberis Recens, Fructus Jujubae, Radix Glycyrrhizae.

Summary of the invention

Technical problem to be solved by this invention provides a kind of medicine for the treatment of flu.

Pharmaceutical composition of the present invention, make its active component raw material and comprise following components in weight percentage: 4～8 parts of Radix Puerariaes, 2～4 parts in Herba Ephedrae, 2～4 parts of Ramulus Cinnamomi, 2～4 parts of the Radix Paeoniae Albas, 2～4 parts of Semen Armeniacae Amarums add the preparation that acceptable accessories or complementary composition are prepared from.

Further, pharmaceutical composition of the present invention, make its active component raw material and also comprise following components in weight percentage: 0.5～3 part in Rhizoma Zingiberis Recens, 1～5 part in Fructus Jujubae, 1～3 part in Radix Glycyrrhizae add the preparation that acceptable accessories or complementary composition are prepared from.

Wherein, above-mentioned preparation is an oral formulations, can be oral liquid, tablet, capsule, granule etc.

Another technical problem to be solved by this invention provides preparation of drug combination method of the present invention.

When pharmaceutical composition of the present invention is that active component is used for the treatment of flu and the time can reaches therapeutic purposes with Radix Puerariae, Herba Ephedrae, Ramulus Cinnamomi, the Radix Paeoniae Alba, Semen Armeniacae Amarum.With Radix Puerariae, Herba Ephedrae, Ramulus Cinnamomi, the Radix Paeoniae Alba, Semen Armeniacae Amarum, Rhizoma Zingiberis Recens, Fructus Jujubae, Radix Glycyrrhizae eight flavor raw materials is that the active component compatibility is when using, can reach best concerted application of drugs, wherein Radix Puerariae, Herba Ephedrae are monarch drug, and Ramulus Cinnamomi, the Radix Paeoniae Alba, Semen Armeniacae Amarum are ministerial drug, Rhizoma Zingiberis Recens, Fructus Jujubae are adjuvant drug, and Radix Glycyrrhizae is a messenger drug.Therapeutic effect is clear and definite, and the analgesic drug effect of cold expelling is remarkable, is the clinical treatment flu, and especially wind and cold diseases caused by exogenous pathogenic factor type flu provides a kind of new selection.

The specific embodiment

Pharmaceutical composition of the present invention is made its active component raw material and is comprised following components in weight percentage: 4～8 parts of Radix Puerariaes, 2～4 parts in Herba Ephedrae, 2～4 parts of Ramulus Cinnamomi, 2～4 parts of the Radix Paeoniae Albas, 2～4 parts of Semen Armeniacae Amarums.

Further preferred pharmaceutical composition of the present invention is made its active component raw material and is comprised following components in weight percentage: 6 parts of Radix Puerariaes, 3 parts in Herba Ephedrae, 3 parts of Ramulus Cinnamomi, 3 parts of the Radix Paeoniae Albas, 3 parts of Semen Armeniacae Amarums.

Preferred further, on the basis of above-mentioned raw materials, to add Rhizoma Zingiberis Recens, Fructus Jujubae, Radix Glycyrrhizae again and can make efficacy stability, auxiliary five tastes raw material performance treatment flu reaches antibiotic, antiviral, analgesic, diaphoresis, analgesic therapeutical effect.

Wherein the weight proportion of Rhizoma Zingiberis Recens, Fructus Jujubae, Radix Glycyrrhizae is: 0.5～3 part in Rhizoma Zingiberis Recens, 1～5 part in Fructus Jujubae, 1～3 part in Radix Glycyrrhizae.

Further preferred pharmaceutical compositions is made its active component raw material and is also comprised the following components in weight percentage composition: 2 parts in Rhizoma Zingiberis Recens, 3 parts in Fructus Jujubae, 2 parts in Radix Glycyrrhizae.

The present invention treats the cold medicine compositions, and it is to be main active with Radix Puerariae, Herba Ephedrae, Ramulus Cinnamomi, the Radix Paeoniae Alba, Semen Armeniacae Amarum, adds the preparation that acceptable accessories or complementary composition are prepared from.Can reach therapeutic purposes when using above-mentioned five tastes material combination treatment to catch a cold, also can be according to clinical needs on the basis of this five tastes raw material, an amount of plus-minus.Adopt Rhizoma Zingiberis Recens, Fructus Jujubae, Radix Glycyrrhizae as the present invention.

Wherein, above-mentioned preparation is an oral formulations, comprises oral liquid, tablet, capsule, granule etc.

Cold medicine compositions of the present invention is made up of eight flavor raw materials, and wherein Radix Puerariae, Herba Ephedrae are monarch drug, and Ramulus Cinnamomi, the Radix Paeoniae Alba, Semen Armeniacae Amarum are ministerial drug, and Rhizoma Zingiberis Recens, Fructus Jujubae are adjuvant drug, and Radix Glycyrrhizae is a messenger drug.The Northern Qi Dynasty, Xu Zhicai was said in " ten doses ": " this is real with the apoplexy table for expelling the excess with mild diaphoretic, the genus of Radix Puerariae, Herba Ephedrae." Radix Puerariae, Gan Xinping, expelling pathogenic factors from muscles is delivered, analgesic promoting the production of body fluid; Herba Ephedrae, Xin Wen, relieving the exterior syndrome by diaphoresis, the lung qi dispersing relieving cough and asthma is we's monarch drug.Ramulus Cinnamomi, Xin Ganwen, dispelling cold by warming the meridian helps the Herba Ephedrae relieving the exterior syndrome by diaphoresis; The Radix Paeoniae Alba, picric acid is slightly cold, and returns Liver Channel, yin fluid astringing and battalion, both can with the Ramulus Cinnamomi compatibility, harmonizing yingfen and weifen, expelling pathogenic factors from muscles is delivered, again can with Radix Glycyrrhizae compatibility relieving spasm to stop pain, strengthen the merit of Radix Puerariae expelling pathogenic factors from muscles pain relieving jointly; Semen Armeniacae Amarum, bitter temperature helps Herba Ephedrae lung qi dispersing relieving cough and asthma; Rhizoma Zingiberis Recens, hot tepor, expelling cold to relieve vomiting; Fructus Jujubae, sweet flat, support the spleen tonifying YIN; And help osmanthus, Chinese herbaceous peony altogether, Yi Heying defends.The Radix Glycyrrhizae coordinating the actions of various ingredients in a prescription.Because of square epheday intermedia, Semen Armeniacae Amarum are the lung meridian medicine, so need not to add again the messenger drug of using tying-in.

Another technical problem to be solved by this invention provides preparation of drug combination method of the present invention, and it comprises the steps:

A, take by weighing following components in weight percentage: 4～8 parts of Radix Puerariaes, 2～4 parts in Herba Ephedrae, 2～4 parts of Ramulus Cinnamomi, 2～4 parts of the Radix Paeoniae Albas, 2～4 parts of Semen Armeniacae Amarums, 0.5～3 part in Rhizoma Zingiberis Recens, 1～5 part in Fructus Jujubae, 2～4 parts in Radix Glycyrrhizae;

B, get Herba Ephedrae, Ramulus Cinnamomi, Semen Armeniacae Amarum, Rhizoma Zingiberis Recens and extract Aromatic water.Aqueous solution after distillation device is in addition collected, and medicinal residues are standby;

C, get Radix Puerariae, the Radix Paeoniae Alba, Fructus Jujubae, Radix Glycyrrhizae, decoct with water, filter, filtrate I and medicinal residues are standby;

D, step c gained medicinal residues and step b gained medicinal residues merge, decoct with water, filter, aqueous solution and step c gained filtrate I after gained filtrate II and the distillation of step b gained merge, and are concentrated into the clear paste of relative density 1.05-1.12 (80 ℃), cooling, adding ethanol makes and contains alcohol amount and reach 60～75%, leave standstill, filter, get filtrate II I;

E, get steps d filtrate II I, step b gained Aromatic water, add acceptable auxiliary or complementary composition is prepared into oral formulations according to the conventional method on the galenic pharmacy.

The present invention also provides pharmaceutical composition of the present invention to have purposes in the medicine of bacteriostasis in preparation.

Beneficial effect by following test of pesticide effectiveness proof pharmaceutical composition of the present invention.

The used pharmaceutical composition of the present invention of the test of pesticide effectiveness is got by following method preparation, and pharmaceutical composition of the present invention is called for short pharmaceutical composition R.

A, take by weighing following components in weight percentage: 6 parts of Radix Puerariaes, 3 parts in Herba Ephedrae, 3 parts of Ramulus Cinnamomi, 3 parts of the Radix Paeoniae Albas, 3 parts of Semen Armeniacae Amarums, 2 parts in Rhizoma Zingiberis Recens, 3 parts in Fructus Jujubae, 2 parts in Radix Glycyrrhizae;

B, get Herba Ephedrae, Ramulus Cinnamomi, Semen Armeniacae Amarum, Rhizoma Zingiberis Recens and extract Aromatic water.Aqueous solution after distillation device is in addition collected, and medicinal residues are standby;

C, get Radix Puerariae, the Radix Paeoniae Alba, Fructus Jujubae, Radix Glycyrrhizae, decoct with water, filter, filtrate I and medicinal residues are standby;

D, step c gained medicinal residues and step b gained medicinal residues merge, decoct with water 40 minutes, filter, aqueous solution and step c gained filtrate I after gained filtrate II and the distillation of step b gained merge, and are concentrated into the clear paste of relative density 1.05-1.12 (80 ℃), cooling, adding ethanol makes and contains alcohol amount and reach 65%, leave standstill, filter, get filtrate II I;

E, get steps d filtrate II I and reclaim ethanol after, blend step b gained Aromatic water, leave standstill, filter, filtrate adds water to full dose, makes every milliliter to contain the 3g crude drug, mixing, packing is promptly.

One, antibacterial action

1, extracorporeal bacteria inhibitor test:

Experiment material

(1) be subjected to the reagent thing: pharmaceutical composition R is provided 3g crude drug/ml, lot number 960106 by the Wu Liang Ye Group, Yibin Pharmaceutical Co.,Ltd., Yibin,Sichuan.To be subjected to reagent to add the medicinal liquid that sterile distilled water is mixed with 1.5g crude drug/ml during experiment, use for the serial dilution test.

(2) positive control drug and preparation thereof: positive control drug is the injection sodium ampicillin.Lot number: 951040, pharmacy six factories in Chongqing produce.Be mixed with 128 μ g/ml concentration with sterile distilled water, use for the serial dilution test.

(3) test culture medium: Mueller-Hinton culture medium, nutrient agar.

(4) bacterial strain is used in test: 20 kinds of totally 100 strains all are that in January, 1996 to February is separating obtained from Chengdu city-level hospital clinical specimen, and test is preceding through identifying the bacterial strain of confirming again.

(5) Quality Control bacterial strain: staphylococcus aureus (ATCC25923) and colon bacillus (ATCC25922).

Detection method

(1) detection of MIC:, make the liquid line that contains variable concentrations inspection product and do not have the medicine control tube with two times of serial dilutions with the M.H fluid medium, every pipe 1ml, (antibacterial that nutritional requirement is high is diluted with the M.H culture medium that contains 8% Sanguis Leporis seu oryctolagi).Test strain is made 10 respectively ⁶The bacterium liquid of CFU/ml is inoculated in the aforesaid liquid pipe, every pipe 0.05ml.Putting 37 ℃ and cultivated 18 hours, is minimal inhibitory concentration (MIC) with the lowest concentration of drug of asepsis growth.With with the MIC of method parallel testing positive control medicine sodium ampicillin to antibacterial.

(2) detection of MBC: according to the clinical scope of application of these inspection product, the MBC that has carried out common upper respiratory tract infection bacterium (totally 15 strains) detects.Method is on MIC test basis, draws its pastille culture fluid 0.1ml from each asepsis growth pipe, and separate application is in the Nutrient agar plate surface of pastille not (the high antibacterial of nutritional requirement with the Nutrient agar plate that contains 8% Sanguis Leporis seu oryctolagi).Cultivated 18 hours for 37 ℃, still the lowest concentration of drug of asepsis growth is minimal bactericidal concentration (MBC).With MBC with method parallel testing positive control medicine sodium ampicillin.

(3) quality control: for guaranteeing to detect quality, under the same terms of above-mentioned test, the parallel ATCC reference culture of having used carries out Quality Control to test.Experimental result: ampicillin is respectively 0.06 μ g/ml and 3.0 μ g/ml to the MIC result of reference culture staphylococcus aureus (ATCC25923) and colon bacillus (ATCC25922).Each bacterial strain of experiment usefulness is equal well-grown in the culture medium of no medicine.

Experimental result

(1) pharmaceutical composition R and positive control drug injection sodium ampicillin are to minimal inhibitory concentration (MIC) and the MIC scope of antibacterial: the results are shown in Table 1 and table 2.

(2) pharmaceutical composition R and positive control drug injection sodium ampicillin are to the minimal bactericidal concentration (MBC) of antibacterial: the results are shown in Table 3.

From table 1 and table 2 as seen, extracorporeal bacteria inhibitor test is the result show, pharmaceutical composition R has in various degree bacteriostasis to 20 kind of 100 strain antibacterial, wherein with better to bacteriostasis such as second streptococcus, staphylococcus aureus, first streptococcus, streptococcus pneumoniae, Klebsiella pneumonia, diphtheria corynebacteriums.As seen from Table 3, pharmaceutical composition R has killing action to 8 kind of 17 strain antibacterials such as staphylococcus aureus under higher concentration.

2, pharmaceutical composition R is to bacteriostasis test in the animal body

Experiment material

(1) laboratory animal: mice, Kunming kind, one-level, body weight 18～21g, male and female half and half.Provide the animal quality quality certification by laboratory animal room of State Pharmaceutical Administration Sichuan Province antibiotics industrial research institute: real moving Guan Zhidi 96-30 number of river.

(2) experiment antibacterial: beta hemolytic streptococcus.Separate and get from hospital clinical patient specimen, through Sichuan Prov. College for Training Medical Managerial Personnel's medical test is that microbiology teaching and research room leaf is set gloomy associate senior technologist and separated and identify (being numbered 6148), and cultivates with the broth bouillon that contains Sanguis Leporis seu oryctolagi and with the normal saline diluted for use by requirement of experiment.

(3) experiment medicine

1. be subjected to the reagent thing; Pharmaceutical composition R, lot number 960106, concentration is 3g crude drug/ml (every ampoule 10ml medicinal liquid is equivalent to contain crude drug 30g).Produce by the Wu Liang Ye Group, Yibin Pharmaceutical Co.,Ltd., Yibin,Sichuan.Becoming the medicinal liquid of variable concentrations to supply with the animal gastric infusion with distilled water diluting during experiment uses.

2. positive control medicine: injection sodium ampicillin, lot number 960140, every bottle of content 0.5g.Produce by Chongqing pharmacy six factories.During experiment with sterilized water for injection dissolving and be diluted to variable concentrations and supply with the animal muscle injection and use.

Experimental technique

(1) prognostic experiment: on the basis of external bacteriostatic experiment, select the comparatively responsive beta hemolytic streptococcus of pharmaceutical composition R as the experiment antibacterial.To mouse peritoneal bacterial injection liquid, observed 24 hours, try to achieve and cause that mouse death rate is 80～100% bacterial concentration and consumption, and observe of the protective effect of the medicine of various dose simultaneously infection animal.By prognostic experiment, determine to cause infection, and determined the dosage range of two kinds of medicines with every mouse peritoneal injection of beta hemolytic streptococcus stock solution (undiluted person) 0.5ml.

(2) formal experiment: get 110 of mices, body weight 18～21g, male and female half and half.Be divided into 11 groups at random, 10 every group, each 5 of male and female.Respectively as pharmaceutical composition R and sodium ampicillin each 5 dosed administration groups and 1 matched group (distilled water).5 dosage groups of pharmaceutical composition R (adjacent high and low dose group ratio is 1: 0.8) are respectively 96.0,76.8,61.4,49.2 and 39.3g crude drug/kg/ day, divide gastric infusion every day 2 times; 5 dosage groups of sodium ampicillin (adjacent high and low dose group ratio is 1: 0.7) are respectively 980,686,480,336 and 235mg/kg/ day, divide (both sides) arm muscles drug administration by injection every day 2 times.Each treated animal is in causing bacterial infection prevention in preceding 1 hour administration 1 time.After the administration 1 hour, each was organized every mice and causes bacterial infection by lumbar injection beta hemolytic streptococcus liquid 0.5ml (minimum lethal dose).After animal is caused bacterial infection, the number of animals of each group survival of successive administration treatment 4 days and statistics.Calculate median effective dose (ED by following improvement karber's method formula ₅₀) and standard error (SED ₅₀).

ED ₅₀＝log ^-1[Xm-i(∑p-0.5)]

${\mathrm{SED}}_{50}=2.3i{\mathrm{ED}}_{50}\sqrt{\frac{P-P^2}{n}}$

In the formula: Xm is the logarithm of maximal dose; I is the logarithm of adjacent high and low dose ratio (high dose is a molecule); P is that the animal dis motility rate is with fractional representation; N is number of animals in the group.Animal dead and survival condition see Table 4.

Experimental result

(1) pharmaceutical composition R:

ED ₅₀=78.54 ± 7.83g crude drug/kg (fiducial limit of P=0.95)

(2) positive control medicine-ampicillin:

ED ₅₀=447.1 ± 101.2mg/kg (fiducial limit of P=0.95)

Conclusion: carry out two batches of external bacteriostasis result of the tests at different times and all show, pharmaceutical composition R all has in various degree inhibitory action to the common malignant bacteria of strain surplus 20 kind 100 and fungus; Staphylococcus aureus, table Portugal coccus, second streptococcus, streptococcus pneumoniae, meningococcus, diphtheria corynebacterium, Klebsiella pneumonia, Candida albicans etc. all there is bactericidal action.Infection experimental result to bacterial infection animal oral administration shows; beta hemolytic streptococcus to mouse peritoneal injection minimum lethal dose is caused infection; pharmaceutical composition R irritates stomach (oral) treatment 4 days, when heavy dose of infection animal is had protective effect.With animals survived as observation index, its median effective dose (ED ₅₀) be 78.54 ± 7.83g crude drug/kg (fiducial limit of P=0.95) (this dosage is 52.36 times of adult's dosage).Under same period the same terms, try to achieve the positive control medicine---the median effective dose (ED of sodium ampicillin (broad ectrum antibiotic) administered intramuscular ₅₀) be 447.1 ± 101.2mg/kg (fiducial limit of P=0.95) (this dosage is 6.71 times of adult's dosage).

Two, antivirus action

1, extracorporeal antivirus effect effect test

Experiment material

(1) experiment medicine: tested with first batch of bacteriostasis by reagent thing and positive control medicine.

(2) be subjected to examination virus: adenovirus (AdV) 3 types, 7 types, respiratory syncytial virus (RSV), influenza virus A 1 type, first 3 types are provided by Beijing Shoudu Inst. of Pediatrics and institute of viruses respectively.

(3) HEKC (HEK), Hela cell: provide by test center's preparation of Sichuan Province's virus and Sichuan Province health and epidemic prevention station respectively.

Experimental technique

(1) toxicity test of medicine pair cell

Pharmaceutical composition R and BIAOSHI GANMAO CHONGJI are diluted to 1g/ml respectively with Hanks liquid, then with two kinds of medicines that are diluted to 1g/ml with Hanks liquid respectively from 1: 10-1: 320 make proportional diluted.Above-mentioned two kinds of variable concentrations medicines are got 2 HEK of 0.2ml inoculation and Hela cell respectively, adsorb after 40 minutes, add 2% calf serum lactalbumin and Eagles liquid respectively and keep liquid.Establish 2 HEK and contrasts of Hela cell that do not contain medicine simultaneously.Put 37 ℃ of incubators and cultivated 7 days, day by day the observation of cell toxic reaction.

(2) medicine is to the inactivation test of virus

Get the Adv3 type, the Adv7 type, influenza virus A 1 type, first 3 types, respectively with pharmaceutical composition R and 6 kinds of variable concentrations of BIAOSHI GANMAO CHONGJI (1: 10-1: 320) after the mixed in equal amounts, get 2 HEK cell pipes of above-mentioned each mixed liquor 0.2ml adding (establishing each 2 of virus and cell contrasts simultaneously) absorption respectively after 40 minutes, adding does not contain the lactalbumin of medicine and keeps liquid 0.8ml.Put 37 ℃ of one weeks of incubator, observation of cell pathological changes day by day.Whether Adv is the Adv pathological changes to occur as the deactivation index; Whether influenza virus is the absorption of Cavia porcellus blood cell to occur as the deactivation index.

RSV deactivation: get RSV strain 10 ^-1After directly acting on 40 minutes with pharmaceutical composition R and 6 kinds of variable concentrations medicines of BIAOSHI GANMAO CHONGJI mixed in equal amounts respectively, get above-mentioned each mixed liquor 0.2ml respectively, inoculate 2 Hela cell pipes after the washing of Hanks liquid, (establishing each 2 of virus and cell contrasts simultaneously) absorption is after 40 minutes, add 2% calf serum Eagles liquid 0.8ml, kept flat 35 ℃ of incubators 24 hours, change next day and put 35 ℃ of rotary incubators (12 commentaries on classics/time), every day, observation of cell merged pathological changes, pathological changes whether occurred merging as the deactivation index with cell.

Experimental result

(1) medicine pair cell toxicity test: the results are shown in Table 5.The pharmaceutical composition R of 1: 10 as seen from Table 5 ,～1: 320 concentration and BIAOSHI GANMAO CHONGJI (positive drug) are to HEK and the equal non-toxic reaction of Hela cell.

(2) medicine is tested inactivation of virus: the results are shown in Table 6.As seen from Table 6, pharmaceutical composition R is at 25mg crude drug/more than the ml concentration respiratory syncytial virus (RSV) is had deactivation; When 50 and the 100mg crude drug/ml concentration, respectively adenovirus (Adv) 3 types and 7 types there is deactivation; Susceptible malicious first 1 type of convection current and first 3 types have deactivation respectively when 100 and the 50mg crude drug/ml concentration.BIAOSHI GANMAO CHONGJI (positive drug) has deactivation to influenza virus, and RSV is had the part deactivation, and Adv is not had deactivation.

Table 5 and table 6 Chinese medicine compositions R (abbreviation is subjected to reagent), BIAOSHI GANMAO CHONGJI (abbreviation positive drug).

2, interior resisting virus effect test: to influenza virus A-prime Mus lung adapted strain (FM ₁) the therapeutical effect test of infecting mouse

Experiment material

(1) is subjected to the reagent thing: pharmaceutical composition R, Wu Liang Ye Group, Yibin Pharmaceutical Co.,Ltd., Yibin,Sichuan, lot number 950201.

(2) positive control medicine

1. antivirus oral liquid, the imperial Medicines and Health Product company limited in east, Chinese-foreign joint Tai'an, lot number 940128.

2. virazole (virazole acid injection), Yangzhou, Jiangsu pharmaceutical factory, lot number 950105.

(3) Strain: influenza virus A-prime Mus lung adapted strain (FM ₁Strain), provide by Virology Inst., China Academy of Preventive Medicine Sciences.Date saved 931201, lyophilisation is preserved.

(4) animal: the NIH mice, male and female half and half, 18-20g, Beijing Biological Product Inst. provides, quality certification 9509M23.

(5) Embryo Gallus domesticus: embryo age on the 9th～11, Beijing Biological Product Inst. provides.

(6) penicillin (lot number 940524N), streptomycin (lot number 950710) are North China Pharmaceutical Factory's product.

Experimental technique

(1) influenza virus FM ₁The preparation of strain: with the FM of vacuum lyophilization ₁Strain virus is to contain the normal saline dilution of each 100u/ml of penicillin and streptomycin, and embryo instar chicken embryo allantois on the 9th is gone in inoculation, cultivates and gather in the crops urine routinely, be inoculated in anaerobism and the amphimicrobian culture medium, the asepsis growth person continues to go down to posterity, get that urine does that hemagglutination test to blood clotting titre reaches 1: 640 or more than.This urine is stored in the liquid nitrogen.Infect the NIH mice with collunarium simultaneously, survey the LD of this viral liquid according to a conventional method ₅₀It is 1: 128.

(2) pharmaceutical composition R is to FM ₁The mensuration of strain virus Lung Index of mice infected by Influenza virus: with animal NIH mice, male and female half and half are divided into six groups, i.e. FM at random ₁The strain infected group, the normal control group, pharmaceutical composition R small dose group (for being equivalent to people's dose,equivalent), the heavy dose of group of pharmaceutical composition R (being 5 times of dose,equivalents) antivirus oral liquid group (for 5 times of dose,equivalents of people), virazole group (being people's dose,equivalent).Every treated animal 12-17 only.Normal control and virus group are irritated stomach with normal saline, every day secondary, each 0.5ml.All the other each group is all with the method gastric infusion, and 2h after the medication five times is with 6LD ₅₀Virus quantity 50 μ l collunariums infect the mice through slight etherization, and the normal control group is then with normal allantois drop nose.Metainfective mice continues medication 7 times (shared medicine 12 times), pluck eyeball sacrificed by exsanguination animal after, weigh, pluck lung and claim lung heavy.Calculate each treated animal lung index and lung index suppression ratio by following formula:

(3) pharmaceutical composition R is to FM ₁The influence of virus multiplication in the strain virus infecting mouse lung homogenate: survey the blood clotting titre of clear liquid in the mouse lung homogenate, show the virus multiplication degree.Use the microwell plate method.Get above-mentioned each experimental animal lung, make lung homogenate, the centrifugal 30min of 3000rpm (4 ℃) gets supernatant and does hemagglutination test routinely.

(4) pharmaceutical composition R is to FM ₁The influence that strain virus infecting mouse pneumonopathy becomes: get the lung of above-mentioned each group (5 every group) animal, do check pathological section routinely, observe pneumonopathy range degree.

(5) pharmaceutical composition R is to FM ₁The dead protective effect of strain virus infecting mouse: mice group and route of administration such as preceding, medication is 5 times before infecting, and infects back medication 9 times, shared medicine 7 days.FM ₁Strain virus infective dose 6LD ₅₀(preserving after two months in the liquid nitrogen), 50ul/ Mus, each treated animal death toll of observed and recorded and existence natural law were observed 14 days altogether day by day, calculated the dead protective rate of each treated animal and prolonged survival rate.

Experimental result

(1) pharmaceutical composition R is to FM ₁The influence of strain virus Lung Index of mice infected by Influenza virus the results are shown in Table 7.Know that by table 7 the heavy dose of group of pharmaceutical composition R can alleviate influenza virus FM very significantly ₁Strain is infected and the pulmonary lesion that causes, and the lung index has very significantly and reduces (P＜0.01).

(2) pharmaceutical composition R is to FM ₁The influence of virus titer the results are shown in Table 8 in the strain infecting mouse lung homogenate.Know that by table 8 the large and small dosage group of pharmaceutical composition R all can suppress FM very significantly ₁The propagation of strain virus in mouse lung, the blood clotting titre of virus reduces (P＜0.01) very significantly.

(3) pharmaceutical composition R is to FM ₁The influence of strain infecting mouse pneumonopathy range degree the results are shown in Table 9.Know that by table 9 pharmaceutical composition R is to FM ₁The influenzae strain virus infected mice can alleviate its pulmonary lesion very significantly, and dosage is big, acts on more obvious.

(4) pharmaceutical composition R is to FM ₁Dead protective rate of influenzae strain virus infected mice and the influence that prolongs survival rate the results are shown in Table 10.Know that by table 10 the large and small dosage group of pharmaceutical composition R is to FM ₁The lethal effect of strain virus infecting mouse does not all have significant protective effect, and the life span of infecting mouse is not also had significant prolongation effect.

Conclusion: pharmaceutical composition R has deactivation in 25mg crude drug/ml concentration to respiratory syncytial virus (RSV); In 12.5mg crude drug/ml concentration RSV there is the part deactivation.Pharmaceutical composition R in 50mg crude drug/ml concentration to adenovirus (Adv) 3 types; In 0.1mg crude drug/ml concentration the Adv7 type there is deactivation.Pharmaceutical composition R in 0.1mg crude drug/ml concentration to influenza first 1 type; In 50mg crude drug/ml concentration influenza first 3 types there is deactivation.The result finds out, the anti-RSV of pharmaceutical composition R, Adv3 type, 7 types, and the effect of influenza first 3 type viruses is better than BIAOSHI GANMAO CHONGJI.Confirm that by experimental result in the body pharmaceutical composition R can suppress FM very significantly ₁The propagation of strain virus in the infected mice lung, the effect of large and small dosage is similar; Can reduce the lung index significantly when heavy dose of, the trend that reduces is also arranged when low dose of; Pathological change to infecting mouse lung tissue section shows that the pharmaceutical composition R of large and small dosage all can significantly alleviate the pulmonary lesion of infected mice, thereby shows that pharmaceutical composition R has antivirus action preferably.Be applied to clinically in conjunction with said preparation, viral upper respiratory tract infection and influenza had significant curative effect, so think that stronger antivirus action is arranged.

Three, refrigeration function

1, to the influence of fever in rabbits body temperature due to typhoid fever, paratyphoid fever, the second triple vaccine

Experiment material

(1) experiment medicine

1. be subjected to the reagent thing: pharmaceutical composition R, by Wu Liang Ye Group, Yibin Pharmaceutical Co.,Ltd., Yibin,Sichuan preparation with 3g crude drug/ml, lot number 920305 are provided.

2. positive control medicine: the chill exterior GANMAO CHONGJI, Benxi, Liaoning the 3rd pharmaceutical factory produces, 1g crude drug/g electuary, lot number 911201.

(2) pyrogenicity agent: typhoid fever, paratyphoid fever, second triple vaccine, by Chengdu biological product institute production.

(3) animal: rabbit, one-level, the male and female dual-purpose, body weight 2～2.5kg is provided by Huaxi Medical Univ's Experimental Animal Center, the animal quality certification: real moving Guan Zhidi 92-121 number of river.

Method and result: get 80 of healthy rabbits, male and female are regardless of, put earlier in the laboratory of about 20 ℃ of room temperatures, the conventional raising 3 days, it is conformed, and every day at the upper and lower noon use WM2-03 temperature indicator (Shanghai medical apparatus factory product) to measure each rabbit anus temperature value once respectively, observe each rabbit body temperature situation of change, with the abnormal body temperature person and in one day body temperature to change the greater (above 0.3 ℃) superseded.Formal experiment that morning twice earlier (30min at interval) is measured rabbit anus temperature, selects for use body temperature normal (38.5～39.5 ℃) and twice body temperature difference to be no more than 0.2 ℃ rabbit for usefulness, and with the meansigma methods of this twice body temperature value as its normal value.Each rabbit is injected typhoid fever, paratyphoid fever, second triple vaccine liquid 2ml/kg pyrogenicity by auricular vein then.Measure the anus temperature behind the 1h, select for use fervescence more than 0.6 ℃ the person test.Be divided into 6 groups with the intensification value by Latin square, 10 every group.Gavage medicine by dosage shown in the table 11, per hour measure each rabbit anus temperature after the administration once, continuous 6 hours.Experimental result sees Table 11.

As seen from Table 11, when the fever in rabbit due to the bacterial endotoxin gavages pharmaceutical composition R low dosage (for 5 times of adult's dosage), do not see the trivial solution heat effect, in during dosage (for adult 15 times of dosage), in 2-6 after taking medicine hour refrigeration function is arranged, but the enhanced effect of opinion heat effect not when dosage increases, low dosage BIAOSHI GANMAO CHONGJI in addition, after taking medicine, in 3-6 hour refrigeration function is arranged also, can see through comparative observation, the onset time of pharmaceutical composition R, the watch GANMAO CHONGJI was fast slightly, but the action intensity of two medicines and hold time basic identical, its difference not statistically significant.

2, to the influence of rat fever body temperature due to the beer yeast

Experiment material

(1) experiment medicine:

1. be subjected to the reagent thing: pharmaceutical composition R, by Wu Liang Ye Group, Yibin Pharmaceutical Co.,Ltd., Yibin,Sichuan preparation with 3g crude drug/ml, lot number 920305 are provided.

2. positive control medicine: the chill exterior GANMAO CHONGJI, Benxi, Liaoning the 3rd pharmaceutical factory produces, 1g crude drug/g electuary, lot number 911201.

(2) pyrogenicity agent: beer yeast, Chengdu brewery provides, and fetches back 87 ℃ of oven dry, pulverizes the back and crosses 80 mesh sieves, and dried yeast powder fits in 20% suspension (containing 0.5% sodium carboxymethyl cellulose) with distilled water, and pressure sterilizing 30 minutes is standby.

(3) animal: rat, SD kind, one-level, body weight 150～200g, male and female half and half.Provide the animal quality certification by Sichuan Academy of Medical Sciences's Experimental Animal Center: real moving Guan Zhidi 92-154 number of river.

Experimental technique and result: use WM2-03 temperature indicator (Shanghai Medical Instrument and Meter Factory's product) in pyrogenicity before measurement rat anus temperature twice, 30 minutes at interval.The animal of choosing 37.5～38.5 ℃ of anus temperature is for usefulness, and with the meansigma methods of this twice body temperature as its normal body temperature value.Each rat back subcutaneous injection 20% beer yeast suspension 1ml/100g pyrogenicity (peak value of this pyrogenicity 8h after pyrogenicity reaches), behind pyrogenicity 7.5h, survey each Mus anus temperature, eliminate the intensification value less than 0.8 ℃ of person, the back is divided into 6 groups with the intensification value by Latin square, 10 every group.Gavage medicine by dosage shown in the table 12, administration began thermometric after 1 hour, and continuous 6 hours, experimental result saw Table 12.

As seen from Table 12, the injection beer yeast causes the rat of heating, when gavaging pharmaceutical composition R low dosage (for 5 times of adult's dosage), do not see the trivial solution heat effect, during middle dosage (for 15 times of dosage of adult), the trivial solution heat effect is all arranged in 1-6 hour, and act on enhancing with the dosage increase, and the low dosage BIAOSHI GANMAO CHONGJI does not have the trivial solution heat effect yet, and middle dosage BIAOSHI GANMAO CHONGJI just occurred refrigeration function in 3 hours later on taking medicine.Therefore same visible pharmaceutical composition R is the rapid-action and close characteristics of two medicine intensity than BIAOSHI GANMAO CHONGJI.

Four, perspiration

Experiment material

1 experiment medicine

1. be subjected to the reagent thing: pharmaceutical composition R is provided 3g crude drug/ml, lot number 920305 by the Wu Liang Ye Group, Yibin Pharmaceutical Co.,Ltd., Yibin,Sichuan.

2. positive control medicine: chill exterior GANMAO CHONGJI, Benxi, Liaoning the 3rd pharmaceutical factory, 1g crude drug/g electuary, lot number 911201.

2 animals: rat, the SD kind, one-level, body weight 150～200g, male and female half and half are provided by Sichuan Academy of Medical Sciences's Experimental Animal Center, the animal quality certification: the real moving pipe matter in river 92-154 number.

Experimental technique and result

Choose 40 of healthy SD kind rat, body weight 150～200g, the male and female dual-purpose is divided into 4 groups by the body weight stratified random.With each Mus vola distilled water scrub, gavages by table 13 dosage respectively then and be subjected to the reagent thing before the experiment, or the positive control medicine gavages BIAOSHI GANMAO CHONGJI liquid, negative control group gavages the equal-volume distilled water.50min after the administration in the rat stationary magazine creel of packing into each group rat special, exposes two lower limb respectively, and rest 5min will go into the perspiration that is occurred when struggle in the tube back with dry cotton ball then and be stained with dried gently with dry cotton ball.Afterwards, coat and Tian Yigao wall reagent A liquid respectively at each Mus skin of sole of foot, treat intensive drying after, the very thin again B liquid of coating, and count with the antiperspirant that is occurred in the magnifier observed and recorded 10min.With level Four scoring method evaluation diaphoresis intensity of force, standards of grading are as follows:

Four minutes: the antiperspirant on 5 meat pads of every foot was counted altogether more than 150, and antiperspirant point is intensive, merged; Three minutes: antiperspirant was counted at 100～150; Two minutes: antiperspirant was counted 50～100; One minute: antiperspirant was counted below 50, is dispersed in.Experimental result such as table 13.

Table 13 is the result show: pharmaceutical composition R 5 times of people's consumptions, 15 times (7.5, under the condition of 22.5g crude drug/kg), can obviously promote normal rat foot sole of the foot portion sweat gland secretion, illustrate that this is subjected to reagent thing and positive control medicine that perspiration is all arranged, and certain dose-effect relationship is arranged, under the Isodose, be subjected to the reagent thing to be better than the positive control medicine.

Five, antiinflammatory action

1, to the influence (sufficient volumetric method) of rat Ovum Gallus domesticus album foot swelling

Experiment material

(1) experiment medicine

1. be subjected to the reagent thing: pharmaceutical composition R is provided 3g crude drug/ml, lot number 920305 by the Wu Liang Ye Group, Yibin Pharmaceutical Co.,Ltd., Yibin,Sichuan

2. positive control medicine: chill exterior GANMAO CHONGJI, Benxi, Liaoning the 3rd pharmaceutical factory, 1g crude drug/g electuary, lot number 911201.

(2) animal: rat, SD kind, one-level, body weight 150～200g, male and female half and half.Provide the animal quality certification by Sichuan Academy of Medical Sciences's Experimental Animal Center: real moving Guan Zhidi 92-154 number of river.

Experimental technique and result: rat is divided into 6 groups at random by sex, body weight, and early stage is measured the right back sufficient volume twice of every Mus in experiment, and the record meansigma methods is a sufficient volume under the normal condition.Then respectively at cause scorching before 24h, 12h, 1h gavages medicine by table 14 dosage.1h causes inflammation with the only sufficient plantar subcutaneous injection of 10% fresh Ovum Gallus domesticus album normal saline solution 0.1ml/ after the last administration, causes the scorching every 30min in back and measures sufficient volume once, continuous 5 times, the results are shown in Table 14.

The result finds out from table 14, and low dose pharmaceutical compositions R does not have the obvious anti-inflammatory and anti effect, and the pharmaceutical composition R of middle and high dosage has stronger antiinflammatory action, and certain dose-effect relationship is arranged.Its antiinflammatory characteristics are that onset is fast than BIAOSHI GANMAO CHONGJI, hold time for a long time.The BIAOSHI GANMAO CHONGJI of low dosage does not have the obvious anti-inflammatory and anti effect yet.

2, the influence of xylol induced mice auricle inflammation

Experiment material

(1) is subjected to reagent thing and positive control medicine and preparation thereof: test with diaphoresis.

(2) proinflammatory agent: dimethylbenzene, AR, chemical reagent factory in Beijing produces

(3) animal: mice, Kunming kind, one-level, body weight 18～22g, male and female half and half.Provide the animal quality certification by Huaxi Medical Univ's Experimental Animal Center: real moving Guan Zhidi 92-203 number of river.

Experimental technique and result: mice is divided into 6 groups at random, pressed the successive administration of dosage shown in the table 15 3 days.Melted paraxylene is 3 in the 40min after the last administration, each Mus auris dextra exterior feature.Put to death animal behind the 30min, clip two ears take out auricle with the 8mm card punch, and torsion balance is weighed, and represents inflammation degree, result such as table 15 with left and right sides auricle weight difference.

The result finds out from table 15, and (the 45g crude drug/kg) pharmaceutical composition R xylol induced mice inflammation has certain inhibitory action to high dose; (7.5g crude drug/kg), middle dosage are (though pharmaceutical composition R of 22.5g crude drug/kg) and BIAOSHI GANMAO CHONGJI are to this performance inhibition trend, not statistically significant for low dosage.

3, to the influence of mouse peritoneal capillary permeability

Experiment material

(1) experiment medicine: pharmaceutical composition R is provided 3g crude drug/ml, lot number 920305 by the Wu Liang Ye Group, Yibin Pharmaceutical Co.,Ltd., Yibin,Sichuan

2. positive control medicine: chill exterior GANMAO CHONGJI, Benxi, Liaoning the 3rd pharmaceutical factory, 1g crude drug/g electuary, lot number 911201.

(2) animal: mice, Kunming kind, 18～22g, male and female half and half.Provide the animal quality certification by Huaxi Medical Univ's Experimental Animal Center: real moving Guan Zhidi 92-203 number of river.

Experimental technique and result: mice is divided into 6 groups at random, pressed the successive administration of dosage shown in the table 16 3 days.30min after the last administration, each caudal vein only inject 2% AZO-blue normal saline solution 0.1ml/, and lumbar injection 0.6% acetum 0.2ml/ simultaneously.Put to death animal behind the 20min, wash the abdominal cavity repeatedly with the 5ml distilled water, the centrifugal 5min of eluate 1000rpm/min, the 590nm colorimetric with OD value reflection mouse peritoneal capillary permeability situation of change, the results are shown in Table 16.

The result finds out from table 16: the pharmaceutical composition R of dosage the same with BIAOSHI GANMAO CHONGJI, all have and suppress the effect of mouse peritoneal capillary permeability more significantly.Through comparing as seen, pharmaceutical composition R is better than BIAOSHI GANMAO CHONGJI.

Brief summary: warp is tested the foot swelling of rat Ovum Gallus domesticus album, xylol induced mice auricle inflammation test, and Dichlorodiphenyl Acetate induced mice abdominal cavity capillary permeability increases experiment, confirms that all pharmaceutical composition R has stronger antiinflammatory action.

Six, analgesic activity

1, to the influence (acetic acid causes mouse writhing method) of mice pain reaction

Experiment material

(1) be subjected to reagent thing and positive control medicine and preparation thereof: with analgesic experiment, morphine hydrochloride injection (0.1%).

(2) animal: mice, the Kunming kind, one-level, body weight 18～22g, male and female half and half are provided by Huaxi Medical Univ's Experimental Animal Center, the animal quality certification: real moving Guan Zhidi 92-203 number of river.

(3) algogen: glacial acetic acid, produce by Chengdu chemicals factory

Experimental technique and result: mice is divided into 7 groups by sex, body weight stratified random, and except that positive control medicine morphine hydrochloride (5mg/kg) ip, all the other press the gastric infusion of dosage shown in the table 17.30min after the administration, each Mus ip 0.7% glacial acetic acid 0.2ml/ only, in the record 20min animal turn round body number (writhing response: mice stretches back leg, and abdominal part shrinks indent, and buttocks is raised, the body distortion), result of the test sees Table 17.

The result finds out from table 17, and high dose medicament compositions R Dichlorodiphenyl Acetate induced mice pain has certain analgesic effect, and low, middle dose drug compositions R and BIAOSHI GANMAO CHONGJI effect are not obvious.

2, to the influence (hot plate stimulus method) of mice pain reaction

Experiment material

(1) is subjected to reagent thing and positive control medicine and preparation thereof: with analgesic experiment.

(2) animal: mice, the Kunming kind, one-level, body weight 18～22g, female entirely, provide the animal quality certification by Huaxi Medical Univ's Experimental Animal Center: real moving Guan Zhidi 92-203 number of river.

Method and result: female mice is surveyed the two subnormal threshold of pains earlier on 55 ± 0.5 ℃ of hot plates, each 30min at interval chooses the animal of the normal threshold of pain in 10～30S and is divided into 6 groups at random, shows the dosage gastric infusion by table 18.After the administration 30,60,90,120min surveys each Mus threshold of pain once, (60S does not lick the metapedes person, the threshold of pain is in 60S) calculated and respectively organized the threshold of pain and improve percentage rate, the results are shown in Table 18.

The result finds out from table 18, and middle dose drug compositions R and chill exterior GANMAO CHONGJI all have significant analgesic activity, and high dose medicament compositions R effect is the strongest, and low dose pharmaceutical compositions R and BIAOSHI GANMAO CHONGJI do not have obvious analgesic activity.

Seven, antitussive effect

1, mice ammonia is drawn coughs EDT ₅₀Influence

Experiment material

(1) is subjected to reagent thing and positive control medicine and preparation thereof: with analgesic experiment.CHUANBEI ZHIKE TANGJIANG is produced by Sichuan Aba pharmaceutical factory.

(2) animal: mice, the Kunming kind, one-level, body weight 18～22g, male and female half and half are provided by Huaxi Medical Univ's Experimental Animal Center, the animal quality certification: real moving Guan Zhidi 92-203 number of river.

Method and result: mice is divided into 7 groups at random, presses the gastric infusion of dosage shown in the table 19, repeat administration is 1 time behind the 1h, 1h after the last administration puts animal in the sprayer unit, and spray is with the strong aqua ammonia of certain hour, spraying stops, and takes out the observation mice immediately and has or not the cough reaction, adjusts spray time.Obtain mice EDT with sequential method ₅₀, calculate the R value, the results are shown in Table 19.

The result finds out from table 19, the BIAOSHI GANMAO CHONGJI antitussive action a little less than, middle dose drug compositions R has remarkable antitussive action.CHUANBEI ZHIKE TANGJIANG has certain antitussive action.

2, mice ammonia is drawn cough preclinical influence

Experiment material

(1) is subjected to reagent thing and positive control medicine and preparation thereof: draw with ammonia and cough EDT ₅₀Experiment.

(2) animal: mice, the Kunming kind, one-level, body weight 18～22g, male and female half and half are provided by Huaxi Medical Univ's Experimental Animal Center, the animal quality certification: real moving Guan Zhidi 92-203 number of river.

Experimental technique and result: mice is divided into 7 groups at random, presses the gastric infusion of dosage shown in the table 20 1 time.1h after the last administration puts mice in the 250ml wide mouthed bottle, splashes into strong aqua ammonia 50 μ l, seals immediately, and mice splashes into to the time that the cough reaction occurs from ammonia record.The results are shown in Table 20.

The result finds out from table 20, the pharmaceutical composition R of three dosage, and BIAOSHI GANMAO CHONGJI and CHUANBEI ZHIKE TANGJIANG all have significant antitussive effect.Deng the dose drug compositions R there was no significant difference of comparing with BIAOSHI GANMAO CHONGJI.

Eight, phlegm-dispelling functions

1, uses the phenol red excretion metering method of mice trachea is experimentized.

Experiment material

(1) is subjected to reagent thing and positive control medicine and preparation thereof: draw with ammonia and cough EDT ₅₀Experiment.

(2) animal: mice, the Kunming kind, one-level, body weight 18～22g, male and female half and half are provided by Huaxi Medical Univ's Experimental Animal Center, the animal quality certification: real moving Guan Zhidi 92-203 number of river.

(3) phenol red, AR.

Experimental technique and result: mice is divided into 7 groups at random, pressed the gastric infusion of dosage shown in the table 21 5 days, water is can't help in the fasting of 8h animal before the experiment.0.5h after the last administration, each Mus lumbar injection 0.5% phenol red 0.5ml/ are only.0.5h animal is put to death in the back, separates trachea, gets 2ml 5%NaHCO ₃Divide the flushing trachea 4 times, return towards Xian 3-4 at every turn, merge eluate 546nm and survey the OD value, calculate phenol red content.The results are shown in Table 21.

The result finds out from table 21, and pharmaceutical composition R, BIAOSHI GANMAO CHONGJI and the CHUANBEI ZHIKE TANGJIANG of three dosage all have remarkable phlegm-dispelling functions.Deng the dose drug compositions R there was no significant difference of comparing with BIAOSHI GANMAO CHONGJI.

Nine, to the influence of body non-specific immunity

1, mice is engulfed the influence of Congo red function

Experiment material

(1) is subjected to reagent thing and positive drug and preparation thereof: with analgesic experiment.

(2) animal: mice, the Kunming kind, one-level, body weight 18～22g, male and female half and half are provided by Huaxi Medical Univ's Experimental Animal Center, the animal quality certification: real moving Guan Zhidi 92-203 number of river.

(3) Congo red, AR.

Experimental technique and result: mice is divided into 6 groups at random, presses the gastric infusion of dosage shown in the table 22 for three days on end.1h after the last administration improves Congo red by Yan Shi and engulfs laboratory method mensuration mice Congo red phagocytic function, the results are shown in Table 22.

The result finds out from table 22, and the pharmaceutical composition R of three dosage and BIAOSHI GANMAO CHONGJI are engulfed the Congo red function to mice and do not made significant difference.

2, to the mouse red blood cell Immune Effects

Experiment material

(1) is subjected to reagent thing and positive control medicine and preparation thereof: with analgesic experiment.

(2) animal: with mice Congo red phagocytic function is tested.

Experimental technique and result: mice is divided into 6 groups at random, pressed the continuous gastric infusion of dosage shown in the table 23 5 days, 1h gets blood after the last administration, presses the Guo Shi improved method and measures the mouse red blood cell immunologic function, the results are shown in Table 23.

The result finds out from table 23, and high dose medicament compositions R has obvious facilitation to hematid immunity function, and low, middle liquid oral dosage and BIAOSHI GANMAO CHONGJI do not have obvious influence to hematid immunity function.

3, brief summary

Pharmaceutical composition R does not have obvious influence to the function that mice engulfs Congo red, but hematid immunity function is had remarkable facilitation at dosage when being 45g crude drug/kg.

More than surplus nine aspects ten experimental result of kind of experimental technique and index show that pharmaceutical composition R has is significantly antibacterial, antiviral, analgesic, analgesia, antiinflammatory, cough-relieving, eliminate the phlegm and promote effects such as erythrocyte immune.

Bacteriostatic test is the result show, pharmaceutical composition R has inhibition and deactivation preferably to multiple common bacteria of respiratory tract and virus.Experiment shows that laboratory animal gives pharmaceutical composition R, when being equivalent to 5 times of clinical dosage, perspiration clearly, when being equivalent to 15 times of clinical dosage, at resisiting influenza virus FM ₁Strain infects, analgesic, analgesia, antiinflammatory, cough-relieving, eliminate the phlegm etc. and to use, and demonstrated more potent fruit.

The Ministry of Public Health medicine was evaluated center approval and was carried out pharmaceutical composition R II clinical trial phase in February, 1998, and specified certain Affiliated Hospital of the college of traditional Chinese medicine to serve as the clinical research responsible department, and the general hospital that is had the clinical trial condition by other three families is a participant.Four tame hospitals bear the II clinical trial phase of the new Chinese medicine three class medicines of pharmaceutical composition R treatment cold card jointly and observe.According to relevant requirements by the common clinical trial protocol of formulating of each participant, at the beginning of 2000, finished system's clinical observation of example surplus in the of 420, adopt unified table of random number random packet, set up observation group and matched group separately, and give double-blind method and observe, enlarge controlled trial and observe and adopt single blind method, and guaranteed the equilibrium, comparable of two groups of aspects such as the age, sex, disease time, severity extent, main symptom integration, syndrome integration, sign integration, virus detection, WBC before treatment.Study subject is selected, in, west diagnosis and differential diagnosis in tcm have clear and definite standard, has standardization and generally acknowledged property, efficacy evaluation adopts semiquantitative assessment method, and is workable.The result shows: the curative effect of pharmaceutical composition R treatment flu reaches 66.20% at the cure rate of blind method stages 210 example, and obvious effective rate reaches 21.90%, and effective percentage reaches 8.57%, and inefficiency is 3.33%, and total obvious effective rate reaches 88.10%, and total effective rate reaches 96.67%; Cure rate in general curative effect 300 examples reaches 65.00%, and obvious effective rate reaches 25.00%, and effective percentage reaches 7.33%, and inefficiency is 2.67%, and total obvious effective rate reaches 90.00%, and total effective rate reaches 97.33%; And the matched group cure rate reaches 50.83%, and obvious effective rate reaches 28.33%, and effective percentage reaches 11.67%, and inefficiency is 9.17%, and total obvious effective rate reaches 79.17%, and total effective rate reaches 90.83%.And observation group all is better than matched group, and p＜0.01 or p＜0.05 illustrates that the curative effect of pharmaceutical composition R is better than matched group.And to the body temperature before and after the treatment change, body temperature fall time, the variation of severity extent, main symptom integration, syndrome integration, sign integration, main symptom changes (aversion to wind and cold, heating, the nasal obstruction watery nasal discharge, stiffness of the nape, sneeze, cough, headache, whole body is miserable), tcm syndrome changes (lossless, thirsty, cough up phlegm, picture of the tongue, tongue fur, pulse condition), sign changes and (swallows red, pharyngeal lymph node folliculus) and reduce the improvement effect that neutrophilic granulocyte classification in the routine blood test before and after the treatment all has significance, p＜0.05 or p＜0.01.Especially the body temperature before and after treatment change, body temperature fall time, sign integration, and heating, the nasal obstruction watery nasal discharge, stiffness of the nape, thirsty, swallow on the curative effect of aspect such as red and all obviously be better than matched group, p＜0.05 or p＜0.01.

Table 1 pharmaceutical composition R is to the minimal inhibitory concentration (MIC) of antibacterial

Bacterium name (bacterium number)	Pharmaceutical composition R (the g crude drug/ml)	Positive control drug (μ g/ml)	Bacterium name (bacterium number)	Pharmaceutical composition R (the g crude drug/ml)	Positive control drug (μ g/ml)
						( 6101 ) ( 6102 ) ( 6103 ) ( 6104 ) ( 6105 ) ( 6106 ) ( 6107 ) ( 6108 ) ( 6109 ) ( 6110 ) ( 6121 ) ( 6122 ) ( 6123 ) ( 6124 ) ( 6125 ) ( 6126 ) ( 6127 ) ( 6128 ) ( 6129 ) ( 6130 ) ( 6131 ) ( 6132 ) ( 6133 ) ( 6134 ) ( 6135 ) ( 6136 ) ( 6137 ) ( 6138 ) ( 6139 ) ( 6140 ) ( 6141 ) ( 6142 ) ( 6143 ) ( 6144 ) ( 6145 ) ( 6146 ) ( 6221 ) ( 6222 ) ( 6223 ) ( 6231 ) ( 6232 ) ( 6233 )	0.38 0.19 0.10 0.38 0.19 0.19 0.38 0.10 0.38 0.19 0.19 0.38 0.38 0.19 0.19 0.38 0.38 0.38 0.38 0.19 0.10 0.19 0.19 0.10 0.19 0.10 0.19 0.19 0.10 0.19 0.10 0.05 0.05 0.19 0.19 0.10 0.38 0.75 0.75 0.38 0.19 0.19	0.25 0.06 0.06 0.25 0.13 0.13 0.25 0.06 0.25 0.13 0.25 0.25 0.50 0.13 0.25 0.50 0.25 0.25 0.25 0.13 0.50 0.50 0.25 0.50 0.50 0.25 0.50 0.25 0.25 0.50 0.13 0.13 0.13 0.25 0.13 0.13 16.00 8.00 8.00 4.00 4.00 2.00	( 6147 ) ( 6148 ) ( 6149 ) ( 6150 ) ( 6151 ) ( 6152 ) ( 6153 ) ( 6154 ) ( 6155 ) ( 6156 ) ( 6157 ) ( 6158 ) ( 6159 ) ( 6160 ) ( 6161 ) ( 6162 ) ( 6163 ) ( 6164 ) ( 6165 ) ( 6171 ) ( 6172 ) ( 6173 ) ( 6174 ) ( 6175 ) ( 6176 ) ( 6177 ) ( 6181 ) ( 6182 ) ( 6183 ) ( 6191 ) ( 6192 ) ( 6211 ) ( 6212 ) ( 6213 ) ( 6214 ) ( 6215 ) ( 6271 ) ( 6272 ) ( 6281 ) ( 6282 ) ( 62101 ) ( 62102 )	0.10 0.05 0.19 0.10 0.38 0.10 0.38 0.19 0.19 0.10 0.10 0.38 0.10 0.10 0.38 0.38 0.75 0.75 0.38 0.38 0.19 0.75 0.75 0.38 0.38 0.19 0.19 0.10 0.10 0.19 0.38 0.38 0.19 0.19 0.38 0.19 0.38 0.38 0.19 0.38 0.10 0.19	0.25 0.13 0.25 0.13 0.50 0.25 0.25 0.25 0.13 0.13 0.13 0.25 0.13 0.25 1.00 0.50 1.00 0.50 0.50 0.50 0.50 1.00 1.00 1.00 0.50 0.50 0.50 0.25 0.25 0.25 0.50 2.00 4.00 2.00 4.00 2.00 16.00 8.00 16.00 32.00 ＞128 ＞128

Continuous table 1

Bacterium morgani (6241) bacterium morgani (6242) bacterium morgani (6243) proteus mirabilis (6251) proteus mirabilis (6252) proteus mirabilis (6253) Bacillus foecalis alkaligenes (6261) Bacillus foecalis alkaligenes (6262)

0.19 0.38 0.38 0.19 0.19 0.19 0.38 0.19

2.00 4.00 4.00 2.00 4.00 4.00 16.00 8.00

Pneumonia gram Salmonella (62103) pseudomonas aeruginosa (62111) pseudomonas aeruginosa (62112) pseudomonas aeruginosa (62113) pseudomonas aeruginosa (62114) Candida albicans (62121) Candida albicans (62122) Candida albicans (62123)

0.19 0.75 0.38 0.38 0.75 0.75 1.50 0.75

＞128 ＞128 ＞128 ＞128 ＞128 ＞128 ＞128 ＞128

Annotate: various antibacterials are well-grown in the culture medium of no medicine.

Table 2 pharmaceutical composition R is to the scope of antibacterial minimal inhibitory concentration (MIC)

Bacterium name (strain number)	Pharmaceutical composition R (the g crude drug/ml)	Positive control drug (μ g/ml)
			Cloudy bacillus (2) the pneumonia gram of staphylococcus aureus (10) table Portugal coccus (10) first streptococcus (10) second streptococcus (10) pneumococcus (10) meningococcus (5) diplococcus siccus (7) corynebacterium diphtheriae (3) diphtheroid (2) Escherichia coli (5) aerobacteria (3) proteus vulgaris (3) bacterium morganis (3) proteus mirabilises (3) Bacillus foecalis alkaligenes (2) enterobacter cloacaes (2) nitre Salmonella (3) pseudomonas aeruginosa (4) Candida albicans (3)	0.10-0.38 0.19-0.38 0.10-0.19 0.05-0.19 0.10-0.38 0.38-0.75 0.19-0.75 0.10-0.19 0.19-0.38 0.19-0.38 0.38-0.75 0.19-0.38 0.19-0.38 0.19 0.19-0.38 0.38 0.19-0.38 0.100.19 0.38-0.75 7.5-1.50	0.06-0.25 0.13-0.50 0.25-0.50 0.13-0.25 0.13-0.50 0.50-1.00 0.50-1.00 0.25-0.50 0.25-0.50 2.00-4.00 8.00-16.00 2.00-4.00 2.00-4.00 2.00-4.00 8.00-16.00 8.00-16.00 16.00-32.00 ＞128 ＞128 ＞128

Table 3 pharmaceutical composition R is to the minimal bactericidal concentration (MBC) of antibacterial

Bacterium name (bacterium number)	Pharmaceutical composition R (the g crude drug/ml)	Positive control drug (μ g/ml)
			Staphylococcus aureus, (6103) staphylococcus aureus, (6109) table Portugal coccus, (6122) table Portugal coccus, (6130) second streptococcus, (6142) second streptococcus, (6144) pneumococcus, (6151) pneumococcus, (6156) meningococcus, (6163) meningococcus, (6165) corynebacterium diphtheriae, (6181) corynebacterium diphtheriae, (6182) pneumonia gram Salmonella, (62101) pneumonia gram Salmonella, (62103) Candida albicans, (62121)	0.38 1.50 0.75 0.75 0.19 0.38 0.75 0.38 1.50 1.50 0.75 0.38 0.38 0.75 1.50	0.25 0.50 1.00 0.50 0.25 1.00 1.00 0.50 2.00 1.00 2.00 1.00 ＞128 ＞128 ＞128

Table 4 pharmaceutical composition R is to the therapeutical effect of bacterial infection mice

Medicine	Dosage	Number of animals (only)	Death toll (only)	Survival number (only)
					Pharmaceutical composition R	96.0g crude drug/kg 76.8g crude drug/kg 61.4g crude drug/kg 49.2g crude drug/kg 39.3g crude drug/kg	10 10 10 10 10	2 5 9 10 10	8 5 1 0 0
The positive control medicine	980mg/kg 686mg/kg 480mgkg 336mg/kg 235mg/kg	10 10 10 10 10	2 3 4 6 8	8 7 6 4 2
					Matched group (distilled water)	-	10	10	0

Table 5 pharmaceutical composition R (being subjected to reagent) pair cell toxicity test result

Cell	Medicine	Drug level (the mg crude drug/ml)						The cell contrast
									100	50	25	12.5	6.25	3.13
		HEK	Be subjected to reagent	- -	- -	- -	- -								- -	- -	- -
Positive drug	-							-	-	-	-	-	-
			Hela	Be subjected to reagent	- -	- -	- -							- -	- -	- -	- -
Positive drug	-	-						-	-	-	-	-

Annotate :-expression medicine pair cell non-toxic reaction.

Table 6 pharmaceutical composition R (being subjected to reagent) is to the inactivation of virus result of the test

Virus	Medicine	Drug level (the mg crude drug/ml)						The cell contrast	Virus control
										100	50	25	12.5	6.25	3.13
		RSV	Be subjected to reagent	- -	- -	- -	± +									+++ ++	+++ +++	- -	+++ +++
Positive drug	± ±							++ ++	++ ++	+++ +++	+++ +++	+++ +++	- -	+++ +++
			Adv3	Be subjected to reagent	- -	- -	++ ++								+++ +++	+++ +++	+++ +++	- -	+++ +++
Positive drug	++ ++	++ ++						++ ++	+++ +++	+++ +++	+++ +++	- -	+++ +++
				Adv7	Be subjected to reagent	- -	++ ++							+++ +++	+++ +++	+++ +++	+++ +++	- -	+++ +++
Positive drug	++ ++	+ ++	+++ +++					+++ +++	+++ +++	+++ +++	- -	+++ +++
					Influenza first 1	Be subjected to the reagent positive drug	(-) (-)						(+) (+)	(+) (+)	(+) (+)	(+) (+)	(+) (+)	(-) (-)	(+) (+)
Influenza first 3	Be subjected to the reagent positive drug	(-) (-)	(-) (+)	(+) (+)				(+) (+)	(+) (+)	(+) (+)	(-) (-)	(+) (+)

Annotate :+, ++, +++expression cytopathy degree; (-) expression blood is inhaled negative, and-expression cell does not have pathological changes, and (+) expression blood is inhaled positive.

Table 7 pharmaceutical composition R is to FM ₁The influence of strain Lung Index of mice infected by Influenza virus

Group	Drug dose	Number of animals (only)	The lung index (X ± S)	Suppression ratio (%)
					Normal control group FM 1The heavy dose of group of strain infected group pharmaceutical composition R small dose group pharmaceutical composition antivirus oral liquid papova azoles group	00 5ml/kg/ day 25ml/kg/ day 25ml/kg/ day 0.1g/kg/ days	17 12 13 13 15 15	0.639±0.062 1.259±0.136 1.211±0.097 1.114±0.096# 1.184±0.220 0.981±0.119#	- - 3.813 11.517 5.957 22.081

Annotate: with FM ₁The strain infected group relatively.# represents P＜0.01

Table 8 pharmaceutical composition R is to FM ₁The influence of strain infecting mouse lung homogenate viral hemoagglutination titre

Group	Drug dose	Animal (only)	Blood clotting titre loge (X ± SD)
				Normal control group FM 1The heavy dose of group of strain infected group pharmaceutical composition small dose group pharmaceutical composition antivirus oral liquid papova azoles group	00 5ml/kg/ day 25ml/kg/ day 25ml/kg/ day 0.1g/kg/ days	17 12 13 13 15 15	- 7.139±0.479 5.000±0.924 * 4.889±0.884 * 5.889±0.719 * 1.265±0.954 *

Annotate: for FM ₁The strain infected group relatively. ^*Expression P＜0.01

Table 9 pharmaceutical composition R is to FM ₁The influence of strain infecting mouse lung pathology section

Group	Drug dose	Animal (only)	Lesion degree			P
							-	+	++
			Normal control group FM 1The heavy dose of papova azoles of strain infected group pharmaceutical composition R small dose group pharmaceutical composition R group	00 5ml/kg/ day 25ml/kg/ day 0.1g/kg/ days	5 5 5 5 5					5 0 4 5 5	0 3 1 0 0	0 2 0 0 0	- - P＜0.05 P＝0.01 P＝0.01

Annotate: (1) P value is and FM ₁The strain infected group relatively.

(2) (-) expression pulmonary does not have pathological changes.(+) expression becomes the master with bronchopathy, and lung tissue does not have pathological changes.(++) expression lung tissue and bronchus pathological changes counterpoise.

Table 10 pharmaceutical composition R is to FM ₁The influence of the dead protective effect of strain virus infecting mouse

Group	Number of animals (only)	Life span (X ± SD, day)	Prolong survival rate (%)	Death toll (only)	Mortality rate (%)
						FM 1The heavy dose of group of strain infected group pharmaceutical composition R small dose group pharmaceutical composition R antivirus oral liquid papova azoles group	16 15 15 16 15	7.31±3.48 7.40±2.85 6.47±3.40 6.31±3.16 10.93±2.74 *	- 1.2 - - 49.7	13 13 13 14 9	81.5 86.7 86.7 87.5 60.0Δ

Annotate: ^*With FM ₁The strain virus infected group is P＜0.01 relatively, Δ and FM ₁The strain virus infected group is P＞0.05. relatively

Table 13 pharmaceutical composition R is to the influence of rat paw portion sweat secretion

Group	Dosage (the g crude drug/kg)	Number of animals (only)	The diaphoresis force intensity (X ± SD)
				Distilled water group pharmaceutical composition R pharmaceutical composition R BIAOSHI GANMAO CHONGJI	Equal-volume 7.5 22.5 7.5	10 10 10 10	1.5±0.53 3.5±0.52 **Δ 3.9±0.31 **Δ 2.8±0.63 **

Annotate: ^*For comparing P＜0.01 with the distilled water group.Δ is to compare P＜0.05 with the BIAOSHI GANMAO CHONGJI group.

Table 14 pharmaceutical composition R is to the influence of rat Ovum Gallus domesticus album foot swelling (X ± SD)

Group	Dosage g crude drug/kg	Number of animals only	Cause scorching back foot swelling volume net added value (mm water column)
								30min	60min	120min	180min	240min
			Distilled water group pharmaceutical composition R group pharmaceutical composition R group pharmaceutical composition R group BIAOSHI GANMAO CHONGJI group BIAOSHI GANMAO CHONGJI group	Equal-volume 7.5 22.5 45 7.5 22.5	10 10 10 10 10 10	20.0±11.0 19.0±8.8 7.1±3.1 **Δ 2.8±2.0 *** 19.4±9.5 13.3±6.7	20.3±11.7 18.8±7.9 5.4±3.3 **Δ 2.4±2.2 *** 19.1±9.4 16.6±6.9						15.0±6.0 14.1±5.8 3.6±3.0 *** 0.4±0.1 *** 14.5±6.0 3.8±3.5 ***	10.6±6.1 10.0±5.5 2.1±1.7 *** 0.1±0.1 *** 10.7±5.4 2.2±2.0 ***	8.4±5.1 7.8±5.1 1.9±1.6 *** 0.0±0.0 *** 8.3±5.3 1.6±1.5 ***

Annotate: 1. compare with the distilled water group: ^*P＜0.01, ^{* *}P＜0.001; 2. compare with positive drug: Δ P＜0.05.

Table 15 pharmaceutical composition R xylol causes the influence (X ± SD) of Mice Auricle inflammation

Group	Dosage (the g crude drug/kg)	Number of animals (only)	Left and right sides auricle weight difference (X ± SD, mg)
				Distilled water group pharmaceutical composition R group pharmaceutical composition R group pharmaceutical composition R group BIAOSHI GANMAO CHONGJI group BIAOSHI GANMAO CHONGJI group	Equal-volume 7.5 22.5 45.0 7.5 22.5	12 12 12 12 12 12	8.72±4.65 6.81±4.03 5.82±3.08 4.15±1.57 ** 7.24±4.65 5.64±3.26

Annotate: compare with the distilled water group: ^*P＜0.01

Table 16 pharmaceutical composition R is to the influence of mouse peritoneal capillary permeability (X ± SD)

Group	Dosage (the g crude drug/kg)	Number of animals (only)	The trap of abdominal cavity washing liquid (X ± SD, OD value)
				Distilled water group pharmaceutical composition R group pharmaceutical composition R group pharmaceutical composition R group BIAOSHI GANMAO CHONGJI group BIAOSHI GANMAO CHONGJI group	Equal-volume 7.5 22.5 45 7.5 22.5	12 12 12 12 12 12	0.71±0.14 0.65±0.12 0.30±0.08 ***Δ 0.56±0.16 * 0.70±0.15 0.39±0.08 ***

Annotate: compare with the distilled water group: ^*P＜0.05, ^{* *}P＜0.001.Compare Δ P＜0.05 with positive drug.

Table 17 pharmaceutical composition R is to the influence (writhing method) of mice pain reaction (X ± SD)

Group	Dosage (the g crude drug/kg)	Number of animals (only)	Incubation period (min)	Turn round the body number (inferior/20min)	Analgesia rate (%)
						Distilled water group pharmaceutical composition R group pharmaceutical composition R group pharmaceutical composition R group BIAOSHI GANMAO CHONGJI group BIAOSHI GANMAO CHONGJI group morphine hydrochloride group	Equal-volume 7.5 22.5 45.0 7.5 22.5 0.005	15 15 15 15 15 15 15	6.21±1.79 7.34±2.68 7.72±4.24 10.97±6.78 * 5.22±1.71 5.40±1.76 20.0±0.0 **	8.57±4.60 7.44±3.28 6.47±2.56 1.93±1.71 ** 8.35±5.26 8.0±5.0 0.0±0.0 ***	- 16.67 24.50 77.48 6.67 6.65 100.0

Annotate: compare with the distilled water group: ^*P＜0.05, ^*P＜0.01, ^{* *}P＜0.001.

Table 19 pharmaceutical composition R draws mice ammonia and coughs EDT ₅₀Influence

Annotate: R＞130%, antitussive action is arranged,＞150% has remarkable antitussive action

Table 20 pharmaceutical composition R draws mice ammonia and coughs preclinical influence (X ± SD)

Group	Dosage (the g crude drug/kg)	Number of animals (only)	Incubation period (S)
				Distilled water group pharmaceutical composition R group pharmaceutical composition R group pharmaceutical composition R group BIAOSHI GANMAO CHONGJI group BIAOSHI GANMAO CHONGJI group fritillary bulb cough syrup	Equal-volume 7.5 22.5 45.0 7.5 22.5 18ml/kg	10 10 10 10 10 10 10	19.7±6.0 34.2±6.8 ** 79.4±12.7 *** 77.8±12.0 *** 31.5±6.1 ** 77.6±12.5 *** 71.9±8.0 ***

Annotate: compare with the distilled water group: ^{* *}P＜0.001.

Table 21 pharmaceutical composition R is to the influence of the phenol red excretion amount of mice trachea

Group	Dosage (the g crude drug/kg)	Number of animals (only)	Phenol red output (μ g/ml) (X ± SD)	Relative percentage (%)
					Distilled water group pharmaceutical composition R group pharmaceutical composition R group pharmaceutical composition R group BIAOSHI GANMAO CHONGJI group BIAOSHI GANMAO CHONGJI group fritillary bulb cough syrup	Equal-volume 7.5 22.5 45.0 7.5 22.5 18ml/kg	lO 10 10 10 10 10 10	1.68±0.8l 4.12±2.04 ** 6.74±3.15 *** 6.36±2.57 *** 3.34±1.86 * 4.27±2.58 ** 4.28±1.44 ***	- 245 401 379 199 254 255

Annotate: compare with the distilled water group, ^*P＜0.05, ^*P＜0.01, ^{* *}P＜0.001.

Table 22 pharmaceutical composition R engulfs the influence (X ± SD) of Congo red function to mice

Group	Dosage (the g crude drug/kg)	Number of animals (only)	Injection Congo red Congo red content (μ g/ml) in the blood in the time of 30 seconds
				Distilled water group pharmaceutical composition R group pharmaceutical composition R group pharmaceutical composition R group BIAOSHI GANMAO CHONGJI group BIAOSHI GANMAO CHONGJI group	Equal-volume 7.5 22.5 45.0 7.5 22.5	12 12 12 12 12 12	10.62±3.36 8.41±3.23 8.81±3.56 11.10±2.93 8.02±2.94 8.96±2.31

Table 23 pharmaceutical composition R is to mouse red blood cell Immune Effects (X ± SD)

Group	Dosage (the g crude drug/kg)	Number of animals (only)	C 3b％	IC％
					Distilled water group pharmaceutical composition R group pharmaceutical composition R group pharmaceutical composition R group BIAOSHI GANMAO CHONGJI group BIAOSHI GANMAO CHONGJI group	Equal-volume 7.5 22.5 45 7.5 22.5	11 11 11 11 11 11	9.45±1.08 9.42±1.90 9.64±2.53 12.55±2.23 ** 9.57±2.11 10.09±2.39	8.14±2.28 8.81±2.34 8.86±2.43 11.09±2.25 ** 7.22±2.28 7.09±2.76

Annotate: compare with the distilled water group: ^*P＜0.01

Table 11 pharmaceutical composition R is to the influence of fever in rabbits body temperature due to the typhoid fever triple vaccine (X ± SD)

Group	Dosage (the g crude drug/kg)	Number of animals (only)	Normal body temperature (℃)	Pyrogenicity body temperature (℃)	The variation of body temperature after the administration (℃)
											1h	2h	3h	4h	5h	6h
					Distilled water group pharmaceutical composition R group pharmaceutical composition R group pharmaceutical composition R group BIAOSHI GANMAO CHONGJI group BIAOSHI GANMAO CHONGJI group	Equal-volume 7.5 22.5 45 7.5 22.5	10 10 10 10 10 10	38.9±0.33 38.8±0.34 38.9±0.29 38.9±0.24 38.9±0.26 38.9±0.21	40.0±0.3 40.1±0.3 40.1±0.4 40.2±0.3 40.1±0.2 40.2±0.3	40.1±0.3 40.1±0.4 40.2±0.3 40.2±0.4 40.1±0.4 40.1±0.3							40.0±0.2 39.9±0.3 39.7±0.3 * 39.6±0.4 * 40.0±0.3 40.0±0.4	40.1±0.3 39.9±0.4 39.7±0.4 * 39.7±0.4 * 39.9±0.5 39.8±0.3 *	40.0±0.4 39.8±0.5 39.6±0.3 * 39.5±0.4 * 39.9±0.4 39.6±0.3 *	39.8±0.3 39.7±0.4 39.5±0.3 * 39.4±0.4 * 39.6±0.5 39.5±0.3 *	39.7±0.4 39.6±0.4 39.2±0.4 * 39.2±0.4 * 39.5±0.5 39.3±0.3 *

Annotate: compare with the distilled water group: ^*P＜0.05.

Table 12 pharmaceutical composition R is to the influence of heating body temperature due to the rat beer yeast (X ± SD)

Group	Dosage (g crude drug kg)	Number of animals (only)	Normal body temperature (℃)	Pyrogenicity body temperature (℃)	The variation of body temperature after the administration (℃)
											1h	2h	3h	4h	5h	6h
					Distilled water group pharmaceutical composition R group pharmaceutical composition R group pharmaceutical composition R group BIAOSHI GANMAO CHONGJI group BIAOSHI GANMAO CHONGJI group	Equal-volume 7.5 22.5 45 7.5 22.5	10 10 10 10 10 10	38.0±0.2 38.0±0.2 38.0±0.2 37.9±0.2 37.9±0.2 37.9±0.1	39.3±0.4 39.4±0.4 39.3±0.4 39.4±0.3 39.4±0.4 39.2±0.4	39.3±0.3 39.1±0.5 38.9±0.4 * 38.9±0.4 * 39.3±0.5 39.1±0.5							39.3±0.4 39.2±0.4 38.8±0.4 * 38.7±0.5 ** 39.3±0.5 39.1±0.5	39.2±0.4 39.0±0.5 38.8±0.4 * 38.6±0.5 ** 39.1±0.4 38.8±0.4 *	39.1±0.3 38.9±0.3 38.6±0.5 * 38.6±0.4 ** 39.0±0.4 38.7±0.4 *	38.9±0.3 38.7±0.4 38.4±0.5 * 38.3±0.5 ** 38.8±0.5 38.5±0.5 *	38.8±0.4 38.7±0.4 38.3±0.5 * 38.2±0.4 ** 38.7±0.5 38.3±0.5 *

Annotate: compare with the distilled water group: ^*P＜0.05, ^*P＜0.01.

Table 18 pharmaceutical composition R to the analgesic activity (hot plate method) of mice (X ± SD, S)

Group	Dosage (the g crude drug/kg)	Number of animals (only)	Pain threshold before the administration	The pain threshold of different time after the administration				Percentage rate is improved in the threshold of pain
												30min	60min	90min	120min	30min	60min	90min	120min
				Distilled water group pharmaceutical composition R pharmaceutical composition R pharmaceutical composition R BIAOSHI GANMAO CHONGJI group BIAOSHI GANMAO CHONGJI group	Equal-volume 7.5 22.5 45 7.5 22.5	10 10 10 10 10 10	21.04±4.23 22.23±4.02 23.59±3.92 23.08±3.97 21.55±4.29 21.70±4.92	25.80±8.97 27.55±8.53 41.80±15.17 ** 55.85±8.13 *** 26.91±9.65 37.10±10.9 *	28.80±17.13 31.24±13.50 49.70±13.35 ** 54.10±13.57 ** 28.38±11.74 37.85±17.65	29.95±7.80 32.23±10.56 43.05±14.18 *** 53.90±12.88 *** 30.73±11.41 40.10±19.34 **	27.70±18.64 30.45±11.27 40.70±15.10 53.80±10.16 ** 30.09±10.21 41.41±18.36									22.62 23.93 77.19 141.98 24.87 70.97	36.88 40.53 110.68 134.40 31.69 74.42	42.35 44.98 82.49 133.54 42.60 84.79	31.65 36.98 72.53 133.10 39.63 90.83

Annotate: compare with the distilled water group: ^*P＜0.05, ^*P＜0.01, ^{* *}P＜0.00

Claims (8)

1. The pharmaceutical composition of 1, treatment flu is characterized in that, makes its active component raw material and comprises following components in weight percentage: 4～8 parts of Radix Puerariaes, 2～4 parts in Herba Ephedrae, 2～4 parts of Ramulus Cinnamomi, 2～4 parts of the Radix Paeoniae Albas, 2～4 parts of Semen Armeniacae Amarums.
2. 2, the pharmaceutical composition of treatment flu according to claim 1 is characterized in that, makes its active component raw material and comprises following components in weight percentage: 6 parts of Radix Puerariaes, 3 parts in Herba Ephedrae, 3 parts of Ramulus Cinnamomi, 3 parts of the Radix Paeoniae Albas, 3 parts of Semen Armeniacae Amarums.
3. 3, the pharmaceutical composition of treatment according to claim 1 flu is characterized in that, it is to be active component with Radix Puerariae, Herba Ephedrae, Ramulus Cinnamomi, the Radix Paeoniae Alba, Semen Armeniacae Amarum, adds the oral formulations that acceptable accessories or complementary composition are prepared from.
4. 4, the pharmaceutical composition of treatment flu according to claim 1 and 2 is characterized in that, makes its active component raw material and also comprises following components in weight percentage: 0.5～3 part in Rhizoma Zingiberis Recens, 1～5 part in Fructus Jujubae, 2～4 parts in Radix Glycyrrhizae.
5. 5, the pharmaceutical composition of treatment flu according to claim 1 and 2 is characterized in that, makes its active component raw material and also comprises following components in weight percentage: 2 parts in Rhizoma Zingiberis Recens, 3 parts in Fructus Jujubae, 2 parts in Radix Glycyrrhizae.
6. 6, the pharmaceutical composition of treatment flu according to claim 4, it is characterized in that, it is to be active component with Radix Puerariae, Herba Ephedrae, Ramulus Cinnamomi, the Radix Paeoniae Alba, Semen Armeniacae Amarum, Rhizoma Zingiberis Recens, Fructus Jujubae, Radix Glycyrrhizae, adds the oral formulations that acceptable accessories or complementary composition are prepared from.
7. 7, a kind of method for preparing the pharmaceutical composition of the described treatment flu of claim 4, it comprises the steps:

   A, take by weighing following components in weight percentage: 4～8 parts of Radix Puerariaes, 2～4 parts in Herba Ephedrae, 2～4 parts of Ramulus Cinnamomi, 2～4 parts of the Radix Paeoniae Albas, 2～4 parts of Semen Armeniacae Amarums, 0.5～3 part in Rhizoma Zingiberis Recens, 1～5 part in Fructus Jujubae, 2～4 parts in Radix Glycyrrhizae;

   B, get Herba Ephedrae, Ramulus Cinnamomi, Semen Armeniacae Amarum, Rhizoma Zingiberis Recens and extract Aromatic water, the aqueous solution after distillation device is in addition collected, and medicinal residues are standby;

   C, get Radix Puerariae, the Radix Paeoniae Alba, Fructus Jujubae, Radix Glycyrrhizae, decoct with water, filter, filtrate I and medicinal residues are standby;

   D, step c gained medicinal residues and step b gained medicinal residues merge, decoct with water, filter, aqueous solution and step c gained filtrate I after gained filtrate II and the distillation of step b gained merge, and are concentrated into the clear paste of relative density 1.05～1.12 (80 ℃), cooling, adding ethanol makes and contains alcohol amount and reach 60～75%, leave standstill, filter, get filtrate II I;

   E, get steps d filtrate II I, step b gained Aromatic water, add acceptable auxiliary or complementary composition is prepared into oral formulations according to the conventional method on the galenic pharmacy.
8. 8, the described pharmaceutical composition of claim 4 has purposes in the medicine of bacteriostasis in preparation.