CN101244170B - Medicament for treating children's exogenous cough and preparation method - Google Patents

Abstract

The invention relates to a drug or a drug composition for treating exogenous cough, in particular to pediatric exogenous cough, as well as the preparation method. The drug is made of the active pharmaceutical ingredients according to the following parts by weight: 5 to 15 portions of herba schizonepetae, 10 to 20 portions of Japanese ardisia herb, 5 to 15 portions of ephedra (honey roast), 1 to 10 portions of bitter almond, 1 to 10 portions of baical skullcap root, 5 to 15 portions of nodakenin, 5 to 15 portions of French pinellia, 5 to 15 portions of pumise, 1 to 10 portions of cicada slough, 1 to 10 portions of tangerine or orange peel, 1 to 10 portions of alkanet and 1 to 10 portions of liquorice root.

Description

The medicine and the preparation method that are used for the treatment of children's exogenous cough

Technical field

The present invention relates to the medicine of a kind of exogenous cough of being used for the treatment of, particularly children's exogenous cough, and the method for preparing it.

Background technology

The exogenous cough is common, the multiple disease of TCM Pediatrics.The exogenous cough, the most common with wind and cold, wind heat syndrome.Because children's's physiological and pathological feature, the card of affection of exogenous wind-cold easily is converted into the cold transformation heat that becomes a common practice and is the wind-cold tightening the lung hot and suffocating card of lung meridian of holding concurrently.The compound Chinese medicinal preparation of this card of treatment is still rare at present.In the compound Chinese medicinal preparation of the children's exogenous cough of listing treatment at present, be that the adaptation patient is in the majority with the syndrome of wind-heat invading lung, wind-cold tightening the lung person takes second place, and serves as that the adaptation patient is still rare with the wind and cold heat-transformation clearly.

Therefore, market to the treatment exogenous cough, especially the compound Chinese medicinal preparation of children's exogenous cough (acute bronchitis) syndrome of heat transformed from wind-cold exists demand.

Summary of the invention

The purpose of this invention is to provide a kind of medicine, be mainly used in treatment exogenous cough, children's exogenous cough especially.

Medicine provided by the invention is made by following materials of weight proportions:

5～15 parts of Herba Schizonepetae, preferred 8～13 parts;

10～20 parts of Herba Ardisiae Japonicaes, preferred 14～22 parts;

5～15 parts in Herba Ephedrae (processed with honey), preferred 7～12 parts;

1～10 part of Semen Armeniacae Amarum, preferred 3～8 parts;

1～10 part of Radix Scutellariae, preferred 3～8 parts;

5～15 parts of Radix Peucedanis, preferred 7～12 parts;

5～15 parts of Rhizoma Pinelliae Preparatum, preferred 7～12 parts;

5～15 parts of Pumexes, preferred 7～14 parts;

1～10 part of Periostracum Cicadae, preferred 3～8 parts;

1～10 part of Pericarpium Citri Reticulatae, preferred 3～8 parts;

1～10 part of Radix Arnebiae (Radix Lithospermi), preferred 3～8 parts;

1～10 part in Radix Glycyrrhizae, preferred 3～8 parts.

Medicine provided by the invention can be made several formulations: any one in tablet, pill, capsule, capsule tablet, powder, granule, suspending agent, gel, the paste.

Another object of the present invention provides a kind of method for preparing said medicine.

The preparation manufacturing method for above mentioned medicine may further comprise the steps:

(1) takes by weighing Herba Ardisiae Japonicae, processed with honey Herba Ephedrae, Rhizoma Pinelliae Preparatum, Semen Armeniacae Amarum, Radix Peucedani, Radix Arnebiae (Radix Lithospermi) by weight, carry out percolation, collect the liquid of filtering, add inclusion agents, the vacuum concentration concentrated solution of winning;

(2) take by weighing Herba Schizonepetae, Pericarpium Citri Reticulatae by weight, use water distillation and extraction, collect volatile oil and filtrate;

(3) claim the medicinal residues of Radix Scutellariae, Pumex, Periostracum Cicadae, Radix Glycyrrhizae and step (1) and (2) gained to merge by weight, the heating decocting is carried suitable number of times and time, collects the water extract, filters, Herba Schizonepetae, Pericarpium Citri Reticulatae extract the filtrate merging of back gained in filtrate and the step (2), concentrate, get second concentrated solution, leave standstill a period of time then, filter, add inclusion agents in gained filtrate, reconcentration gets the 3rd concentrated solution;

(4) get described first concentrated solution and described the 3rd concentrated solution, mix, concentrate, drying is ground into fine powder;

(5) clathrate of forming by described volatile oil and inclusion agents by the proper proportion preparation;

(6) get step (4) fine powder (dry extract) that obtains and the clathrate that step (5) obtains, make suitable dosage form.

Clinical trial shows, medicine provided by the invention is to clinical common children's exogenous cough due to wind and cold, or has wind and cold not separate, and the gesture of heat-transformation is arranged, so that the strongly fragrant and symptoms including cough of heat wrapped by cold, rhinorrhea with clear discharge, and itching throat, or swallow red pharyngalgia thirstyly waits disease that better curative effect is arranged.

The specific embodiment

The exogenous cough of being used for the treatment of provided by the invention, the drug main that is particularly useful for treating children's exogenous cough will be made up of Herba Schizonepetae, Herba Ardisiae Japonicae, Herba Ephedrae (processed with honey), Semen Armeniacae Amarum, Radix Scutellariae, Radix Peucedani, Rhizoma Pinelliae Preparatum, Pumex, Periostracum Cicadae, Pericarpium Citri Reticulatae, Radix Arnebiae (Radix Lithospermi), Radix Glycyrrhizae.To be the inventor propose according to for many years traditional Chinese medical science working experience with to the deep understanding of theory of Chinese medical science this prescription.The function of each main component is as follows in the prescription:

Herba Schizonepetae: nature and flavor suffering, tepor.Return lung, Liver Channel.Function expelling pathogenic wind from the body surface, the rash skin ulcer that disappears, the parch to black hemostasis.Be used for the diseases caused by exogenous pathogenic factor exterior syndrome, cold, fever, headache, cough, laryngopharynx swelling and pain, measles without adequate eruption, the rubella pruritus, skin infection is told the nosebleed hematochezia from the beginning of exterior syndrome is arranged.

Herba Ardisiae Japonicae: have another name called Herba Ardisiae Japonicae (" Chinese medicine voluminous dictionary "), PINGDIMU (" China's book on Chinese herbal medicine ").Acrid in the mouth, little hardship, property is flat.Return lung, Liver Channel.The functionalization phlegm and relieving cough, dampness removing is invigorated blood circulation.Cure mainly new chronic cough and cough, abundant expectoration, sputum mixed with blood, rheumatism arthromyodynia, toxic swelling, dysentery, spitting of blood, haematemesis, impairment caused by overstrain etc.

Herba Ephedrae: acrid in the mouth, little hardship, warm in nature, go into lung, urinary bladder channel.The function expelling cold and relieving exterior syndrome, depressed lung-energy dispersing, diuretic.It is real to cure mainly the typhoid fever table, headache general pain, cough and asthma, edema due to wind pathogen edema, ailment said due to cold or exposure insensitive impediment etc." herbal classic ":: " delivering perspiration, the steam that uncharms, main cough with dyspnea ".

Semen Armeniacae Amarum: bitter in the mouth, slightly warm in nature, slightly poisonous, go into lung, large intestine channel.The function lowering the adverse-rising QI to resolve phlegm, relieving cough and asthma, loosening bowel to relieve constipation.Can be used for multiple cough and asthma disease.

Radix Scutellariae: nature and flavor hardship, cold.Return lung, stomach, gallbladder, large intestine channel.The function heat clearing and damp drying, eliminating fire and detoxication, cooling blood for hemostasis, heat extraction is antiabortive.Be used for the hygropyrexia heat-damp in summer, damp and hot painful abdominal mass is vexed, the jaundice dysentery, and the cough due to lung-heat, the calentura excessive thirst, carbuncle sore tumefacting virus, laryngopharynx swelling and pain, heat in blood is told nosebleed, excessive fetal movement due to heat in pregnancy.

Radix Peucedani: bitter in the mouth, suffering, cold nature.Return lung meridian.The function wind heat clearing away of loosing, lowering the adverse-rising QI to resolve phlegm.Be used for the cough due to pathogenic wind-heat, the expectorant dyspnea of heat type is full, and it is yellow thick to cough up phlegm.。

Rhizoma Pinelliae Preparatum: acrid in the mouth, warm in nature, go into spleen, stomach, lung meridian.Function drying dampness to eliminate phlegm, stopping nausea and vomiting by lowering the adverse flow of QI, dissolving lump and resolving mass.Cure mainly the damp-phlegm cold drink, cough with asthma abundant expectoration, chest and diaphragm distension, vomiting regurgitation, syncope due to accumulation of phlegm headache etc.

Pumex: salty in the mouth, cold in nature.Return lung, kidney channel.The function removing heat from the lung and dissipating phlegm, hard masses softening and resolving.Cure mainly phlegm-heat cough ， Gall tumor, skin ulcer is swollen.

Periostracum Cicadae: sweet in the mouth, salty, cool in nature, go into lung, Liver Channel.The function wind heat of loosing, lung qi dispersing is decided convulsion.Cure mainly affection due to external wind and heat, the cough hoarseness, measles is sent out not smooth thoroughly, rubella pruritus, pediatric epilepsy scared, tetanus etc.。

Pericarpium Citri Reticulatae: acrid in the mouth, hardship, warm in nature, go into spleen, lung meridian.The function regulating QI and lowering the adverseness whets the appetite in the accent, drying dampness to eliminate phlegm.The treatment distension and fullness of the chest and abdomen, anorexia, vomiting hiccup, cough with copious phlegm etc.Modern pharmacological research shows that the contained volatile oil of Pericarpium Citri Reticulatae has the zest phlegm-dispelling functions, and the Pericarpium Citri Reticulatae alcohol extract has strong relexation to guinea-pig isolated trachea.

Radix Arnebiae (Radix Lithospermi): bitter in the mouth, cold in nature.GUIXIN, Liver Channel.Function detoxifcation rash, removing heat from blood and promoting blood circulation.

Radix Glycyrrhizae: sweet in the mouth, property is flat, goes into the heart, lung, spleen, stomach warp.Function and middle emergency, nourishing the lung to arrest cough, detoxifcation, coordinating the actions of various ingredients in a prescription.

More than all medicines cooperate, reach the effect of expelling wind and cold, lung qi dispersing heat clearing away, expelling phlegm for arresting cough jointly, suit with the main syndrome etiology and pathogenesis.And this product is selected survival dose, considers to adapt to disease and children's's physiological and pathological feature everywhere.Card belongs to wind and cold bundle table, then looses based on Xin Wenbiao, but notices that children's is the body of young the moon, can not show to loose too, in order to avoid impairment of body fluid changes; Card is held concurrently hot and suffocating, and still when with Xuan Kaiwei master, to let out consumption clearly with bitter cold lighter though help, and makes a distinction between the important and the lesser one and does not have the side effect of imapirment of the spleen and stomach.This combination meets the main syndrome etiology and pathogenesis, takes into account children's's physiological and pathological feature.Its legislation, select medicine, prescription, meet Chinese medical theory and adapt to the etiology and pathogenesis of disease.

According to the exogenous cough of being used for the treatment of of the present invention, the medicine that is particularly useful for treating children's exogenous cough is to be made by the following weight proportion raw material:

5～15 parts of Herba Schizonepetae, preferred 8～13 parts;

10～20 parts of Herba Ardisiae Japonicaes, preferred 14～22 parts;

5～15 parts in Herba Ephedrae (processed with honey), preferred 7～12 parts;

1～10 part of Semen Armeniacae Amarum, preferred 3～8 parts;

1～10 part of Radix Scutellariae, preferred 3～8 parts;

5～15 parts of Radix Peucedanis, preferred 7～12 parts;

5～15 parts of Rhizoma Pinelliae Preparatum, preferred 7～12 parts;

5～15 parts of Pumexes, preferred 7～14 parts;

1～10 part of Periostracum Cicadae, preferred 3～8 parts;

1～10 part of Pericarpium Citri Reticulatae, preferred 3～8 parts;

1～10 part of Radix Arnebiae (Radix Lithospermi), preferred 3～8 parts;

1～10 part in Radix Glycyrrhizae, preferred 3～8 parts.

In a kind of more preferably embodiment of the present invention, this medicine that is used for the exogenous cough is to be made by the following weight proportion raw material:

9 parts of 10.9 parts of Rhizoma Pinelliae Preparatum of Herba Schizonepetae

10.9 parts of 18 parts of Pumexes of Herba Ardisiae Japonicae

5.5 parts of 9 portions of Periostracum Cicadaes of Herba Ephedrae (processed with honey)

5.5 parts of 5.5 parts of Pericarpium Citri Reticulataes of Semen Armeniacae Amarum

5.5 parts of 5.5 portions of Radix Arnebiae (Radix Lithospermi)s of Radix Scutellariae

5.5 parts in 9 portions of Radix Glycyrrhizaes of Radix Peucedani

Raw material in the said medicine prescription is all medical herbs, and it can be the medical herbs finished product of selling in pharmacy, also can be the medical herbs of new results, for the latter's situation, generally new medical herbs will be dried or is dried, to remove moisture content substantially.

Medicine of the present invention can use separately, perhaps under suitable situation, is used in combination with other drug.

Medicine of the present invention can contain one or more pharmaceutical excipients (carrier).Excipient is added in the ingredient for multiple purpose.This ingredient can provide with unit dosage form easily, and is prepared by methods known in the art.Such method comprises that wherein carrier itself can comprise one or more auxiliary elements with extract component and the combined step of carrier.Usually, the preparation of this dosage form is by extract component and liquid-carrier or powder solid carrier or the combination of the two are mixed mutually, and this product then in case of necessity is shaped.

Pharmaceutical excipient is meant that for the useful excipient of pharmaceutical compositions, it is safe, nontoxic and desirable usually, and comprises the excipient that can be applicable to veterinary and human medicine.This class excipient can be solid, liquid, semisolid, or is gas under the situation that aerosol is formed.

Medicine of the present invention can give with the oral formulations form per os of doing.Medicine of the present invention can give with per os such as tablet, pill, capsule, capsule tablet, powder, granule, suspending agent, gel, pastes.Orally administered composition can comprise common carrier, excipient and diluent.The peroral dosage form of pharmaceutical composition of the present invention can prepare with technology known in the art, and contains the ingredient that effective dose is gone up in treatment.Determine by standard clinical techniques according to working doctor's judgement for the effective oral dose of the treatment of preparation of the present invention.For example, except that the information that medical science handbook and materia medica document provide, can help determine optimal dose with chemical examination in the external or body of knowing.

Preferably, this pharmaceutical preparation is the peroral dosage form of doing.Preferably, this pharmaceutical preparation is solid dosage forms.Preferably, this pharmaceutical preparation is tablet, pill, capsule, capsule tablet, powder and granule.In a specific embodiment of the present invention, this pharmaceutical preparation is granule.The dosage form of doing can comprise the pharmacy acceptable additive, as inclusion agents, excipient, carrier, diluent, stabilizing agent, plasticizer, binding agent, fluidizer, disintegrating agent, filler, lubricant, plasticiser, coloring agent, film former, flavour enhancer, sweeting agent, antiseptic, drug administration carrier (dosing vehicle) and aforementioned in any combination.These all are well-known to those skilled in the art, do not repeat them here.

The example of inclusion agents comprises alpha-cyclodextrin (α-CD), beta-schardinger dextrin-(β-CD) and N-LOK modified starch, preferred beta-schardinger dextrin-.The inclusion agents addition is about 1%～10% of full presciption medicine material total amount.

The example of sweeting agent comprises steviosin, stevioside, protein sugar, xylitol, high fructose, cyclamate etc.These sweeting agents can use separately or use with their two or more form of mixtures.Preferably sucrose or stevioside, the more preferably mixture of the two in the present invention.Can determine to select the consumption of sweeting agent according to the sugariness of dosage form and corresponding sweeting agent, usually 0.01%～0.6%, for example when sugariness was 200～300 times of sucrose, the pharmaceutical solutions consumption was generally 0.07%.

The example of binding agent comprises sodium carboxymethyl cellulose (CMC-Na), low-substituted hydroxypropyl cellulose (L-HPC), Polyethylene Glycol (PEG), polyvinylpyrrolidone (PVP) _K-30, preferably polyethylene ketopyrrolidine (PVP) _K-30

The selection of excipient and consumption by the preparation designer rule of thumb and consider that standard procedure and this area reference material are easy to determine.Solid drugs of the present invention comprises powder, granule, aggregation and compacting component.Preferred route of administering of the present invention is oral.This dosage can provide easily with unit dosage forms, and prepares with any well-known process of pharmaceutical field.Dosage form comprises solid dosage forms, as tablet, pill, powder, capsule, granule, capsule tablet, pouch agent, tablet and lozenge.The particularly preferred dosage form of the present invention is a granule.

The form of paste solution or freeze-dried powder is used for the gastrointestinal tract external administration.Can in powder, add suitable diluent or other pharmaceutical carrier before using and carry out reprovision.Liquid formulations normally cushions, etc. ooze, aqueous solution.The example of suitable diluent has standard normal isotonic saline solution, 5% D/W.This class prescription especially is fit to the gastrointestinal tract external administration, but also can be oral.Preferably add excipient, as polyvinylpyrrolidone, gelatin, oxidized cellulose, arabic gum, Polyethylene Glycol, mannitol, sodium chloride or sodium citrate.

Replacedly, these medicines can be encapsulated, tablet or prepare in Emulsion or syrup, be used for oral.In order to strengthen or stable medicine or help preparation of compositions, can add medicinal solid or liquid-carrier.Liquid-carrier comprises syrup, Oleum Arachidis hypogaeae semen, olive oil, glycerol, saline, alcohols or water.Solid carrier comprises starch, lactose, calcium sulfate, dihydrate, Gypsum Fibrosum powder, magnesium stearate or stearic acid, Pulvis Talci, pectin, arabic gum, agar or gelatin.Carrier also can comprise sustained-release materials, such as glycerol monostearate or the acid of glycerol distearyl, separately or with wax.The amount of solid carrier has nothing in common with each other,

The present invention also provides the method for the aforementioned medicine of preparation, may further comprise the steps:

(1) takes by weighing Herba Ardisiae Japonicae, processed with honey Herba Ephedrae, Rhizoma Pinelliae Preparatum, Semen Armeniacae Amarum, Radix Peucedani, Radix Arnebiae (Radix Lithospermi) by weight, carry out percolation, collect the liquid of filtering, add an amount of inclusion agents, the vacuum concentration concentrated solution of winning;

(2) take by weighing Herba Schizonepetae, Pericarpium Citri Reticulatae by weight, use the water distillation and extraction appropriate time, collect volatile oil and filtrate;

(3) claim the medicinal residues of Radix Scutellariae, Pumex, Periostracum Cicadae, Radix Glycyrrhizae and step (1) and (2) gained to merge by weight, the heating decocting is carried suitable number of times and time, collect the water extract, the filtrate after filtering and extracting with Herba Schizonepetae, Pericarpium Citri Reticulatae merges, and concentrates, get second concentrated solution, leave standstill a period of time then, filter, in filtrate, add inclusion agents, reconcentration gets the 3rd concentrated solution;

(4) get above-mentioned first concentrated solution and the 3rd concentrated solution, mix, concentrate, drying is ground into fine powder;

(5) clathrate of forming by above-mentioned volatile oil and inclusion agents by the proper proportion preparation;

(6) get step (4) fine powder (dry extract) that obtains and the clathrate that step (5) obtains, make suitable dosage form.

In Study on extraction, carried out extraction and the clathrate process condition thereof of Herba Schizonepetae, Pericarpium Citri Reticulatae volatile oil respectively, Six-element medicinal material extract process conditions such as Herba Ardisiae Japonicae preferred, the preferred research of medical material extraction process by water conditions such as Radix Scutellariae, Pumex, Periostracum Cicadae; Concentrate with drying process research in, carried out differently concentrating, drying mode is to the research of dried cream powder melting; In preparations shaping research, preparation prescription, moulding process etc. is studied.By above-mentioned research, the process conditions that are prepared as follows of this product have been determined.

In the step (1), the operation of most preferred percolation can be carried out according to the percolation under Chinese Pharmacopoeia version appendix IO fluid extract in 2000 and the extractum item,, makes solvent with 75% ethanol that is, flooded 24 hours, then in flow velocity (with the 1000g medical material) percolation of per minute 3～5ml.Yet, one of ordinary skill in the art will readily recognize that above condition is done some adjustment, can implement the present invention equally.For example solvent is done some variation, for example adopt the ethanol water of other percentage compositions.We find that in test the ethanol with 60～90% is available.In addition, adopt the methanol aqueous solution or the aqueous acetone solution of suitable concentration also can implement the present invention.In addition, can also be adjusted the time and the flow velocity of dipping.In step (1), concentrated suiting under reduced pressure carried out, and suitable temperature is 40～75 ℃, preferred 50～65 ℃.In the concentration technology research, test of many times show below 60 ℃ ,-0.08MPa vacuum and to be concentrated into relative density for example be that the operating condition of 1.15～1.20 (60 ℃ of heat are surveyed) is preferred.In this step, the inclusion agents addition can be 1～10% scope of full presciption medicine material total amount, preferably in 1.5～7% scope, in 2～5% scope, for example is 2.5% more preferably.

In the step (2), preferably before distillation extraction, be soaked in water a period of time for example 0.5 hour.The addition of water is 4～12 times for Herba Schizonepetae, Pericarpium Citri Reticulatae weight, preferred 6～10 times for example 8 times.Heating distillation extraction 4～10 hours, preferred 6～8 hours, for example 6 hours.

In the step (3), the heating decocting is carried for several times, found through experiments, and the heating decocting is carried three times and can fully be extracted, and amount of water is respectively 8 times, 6 times, 6 times of medical material amount, and extraction time was respectively 1.5 hours, 1 hour, 1 hour.Concentrate preferred 1.04～1.06 (60 ℃ of heat are surveyed) of the relative density of second concentrated solution that obtains with the filtrate merging back after Herba Schizonepetae, the Pericarpium Citri Reticulatae distillation.Time of repose is a few hours, and is preferred more than 12 hours.In this step, the inclusion agents addition preferably is about 2.5% of full presciption medicine material total amount.The relative density of the 3rd concentrated solution is preferably 1.23～1.25 (60 ℃ of heat are surveyed).In a specific embodiment, above-mentioned inclusion agents is a beta-schardinger dextrin-.

In the step (4), concentrated condition can be 1.30～1.32 (60 ℃ of heat are surveyed) for being concentrated into relative density at 60 ℃.The vacuum drying condition for example be 60 ℃ ,-0.08MPa.

In the step (5), can add the volatile oil that obtains in the step (2) of suitable proportion then, mix homogeneously earlier with inclusion agents The suitable solvent such as water furnishing pasty state.As preferably, can be with volatile oil elder generation wiring solution-forming, the alcoholic solution as 50% makes this solution and above-mentioned inclusion agents stick with paste the merging of dress thing then, grinds the suitable time.Final drying obtains volatile oil clathrate compound.In a specific embodiment, can water with beta-schardinger dextrin-furnishing pasty state, in mortar, grind evenly.According to inclusion agents: the volatile oil ratio is (5～15): 1, and took by weighing volatile oil, and used ethanol dilution in preferred 8: 1.Then this diluent and aforementioned paste dress thing are merged grinding.Final drying, pulverizing.Baking temperature is 40～60 ℃, preferred 50 ℃.

In step (6), medicine can be made in tablet, pill, capsule, capsule tablet, powder, granule, suspending agent, gel, the paste any one.For the present invention, preferably make tablet, powder, capsule, granule, more preferably powder, granule, most preferably granule.

In specific embodiments more of the present invention, this medicine is made into granule.In preparation, get the clathrate that fine powder that step (4) obtains and step (5) obtain in required ratio, add an amount of sweeting agent mixing, with binder solution system soft material, sieve, dry at a certain temperature, granulate promptly gets drug particles.But binder solution alcoholic solution form, concentration for example are 3%.Slot size for example is 10～12 orders.

In specific embodiments more of the present invention, this medicine is made into tablet.In preparation, get the clathrate that fine powder that step (4) obtains and step (5) obtain in required ratio, add the polyvinylpolypyrrolidone (PVPP) of an amount of microcrystalline Cellulose and 3%, mixing, 95% alcoholic solution system soft material with 2% 30 POVIDONE K 30 BP/USP 30 (PVP), pushed 16 eye mesh screens, 40～50 ℃ of dryings, granulate, the carboxymethyl starch sodium (CMS-Na) of adding 3% and 0.3% magnesium stearate mixing, compacting is wrapped film-coat in flakes, promptly gets tablet.

Traditional process for preparing medicine is followed in the medicine preparation, with regard to the preparation tablet, relates to grinding, mixing, granulating and compression (when in case of necessity); Or with regard to the preparation hard capsule, relate to grinding, mix and fill.When using liquid-carrier, pharmaceutical preparation is syrup, elixir, Emulsion or aqueous or non-aqueous suspension form.Can directly oral (p.o.) administration or be packed in the soft capsule for this liquid dosage form.

Preparation embodiment

Embodiment 1

Get 10 parts of Herba Ardisiae Japonicaes, 5 parts in processed with honey Herba Ephedrae, 5.2 parts of Rhizoma Pinelliae Preparatum, 1.1 parts of Semen Armeniacae Amarums, 5 parts of Radix Peucedanis, Radix Arnebiae (Radix Lithospermi) and (be weight portion for 1.1 parts, down together), according to the percolation under Chinese Pharmacopoeia version appendix IO fluid extract in 2000 and the extractum item, make solvent with 65% ethanol, flooded 36 hours, then in flow velocity (with the 1000g medical material) percolation of per minute 3～5ml, the collection liquid of filtering, add a certain amount of β-CD (be about full presciption medicine material total amount 1.5%), under 40～75 ℃ and decompression, be concentrated into relative density and be 1.15～1.20 (60 ℃ of heat are surveyed), standby.Get 5 parts of Herba Schizonepetae, 1.1 parts of Pericarpium Citri Reticulataes, add 5 times of water gagings and soaked 0.5 hour, heating distillation extraction 5 hours is collected volatile oil and filtrate, and is standby.Get 1.1 parts of Radix Scutellariaes, 5 parts of Pumexes, 1.1 parts of Periostracum Cicadaes, 1.1 parts in Radix Glycyrrhizae mentions that with above-mentioned alcohol the medicinal residues after the distillation merge, the heating decocting is carried three times, amount of water is respectively 8 times of medical material amount, 6 times, 6 times, extraction time was respectively 1.5 hours, 1 hour, 1 hour, collect three times the water extract, filter and and Herba Schizonepetae, filtrate after the Pericarpium Citri Reticulatae distillation merges, be concentrated into relative density and be about 1.04～1.06 (60 ℃ of heat are surveyed), standing over night (more than 12 hours), get supernatant liquid filtering, in filtrate, add a certain amount of β-CD (be about full presciption medicine material total amount 5%), be concentrated into relative density and be 1.23～1.25 (60 ℃ of heat are surveyed), standby.Get above-mentioned alcohol extraction and water extracting liquid, mix, continuing to be concentrated into relative density is 1.30～1.32 (60 ℃ of heat are surveyed), vacuum drying (60 ℃ ,-0.08MPa), be ground into fine powder, standby.Other gets beta-schardinger dextrin-and places mortar in right amount, adds 4 times of water gagings and grinds evenly, and (beta-schardinger dextrin-: it is an amount of that ratio volatile oil) is got volatile oil in 8: 1, be made into 50% volatile oil ethanol liquid earlier, slowly join then in the mortar, ground 2 hours, in 50 ℃ of dryings, pulverize, standby.Get the Benexate Hydrochloride of above-mentioned dry extract and volatile oil, add an amount of sucrose, steviosin mixing,, cross 10～12 mesh sieves with 3% 30 POVIDONE K 30 BP/USP, 30 alcoholic solution system soft materials, 40 ℃ of dryings, granulate gets granule.

Embodiment 2

Get 20 parts of Herba Ardisiae Japonicaes, 14.7 parts in processed with honey Herba Ephedrae, 14.7 parts of Rhizoma Pinelliae Preparatum, 10 parts of Semen Armeniacae Amarums, 14.7 parts of Radix Peucedanis, 10 parts of Radix Arnebiae (Radix Lithospermi)s, according to the percolation under Chinese Pharmacopoeia version appendix IO fluid extract in 2000 and the extractum item, make solvent with 90% ethanol, flooded 16 hours, then in flow velocity (with the 1000g medical material) percolation of per minute 2～7ml, the collection liquid of filtering, add 2.5% the β-CD be about full presciption medicine material total amount, vacuum concentration is (below 60 ℃,-0.08MPa) be 1.15～1.20 (60 ℃ of heat are surveyed) to relative density, standby.Get 14.7 parts of Herba Schizonepetae, 10 parts of Pericarpium Citri Reticulataes, add 12 times of water gagings and soaked 3 hours, heating distillation extraction 4 hours is collected volatile oil and filtrate, and is standby.Get 10 parts of Radix Scutellariaes, 14.7 parts of Pumexes, 10 parts of Periostracum Cicadaes, 10 parts in Radix Glycyrrhizae mentions that with above-mentioned alcohol the medicinal residues after the distillation merge, the heating decocting is carried three times, amount of water is respectively 8 times of medical material amount, 6 times, 6 times, extraction time was respectively 2 hours, 1.5 hour, 1 hour, collect three times the water extract, filter and and Herba Schizonepetae, filtrate after the Pericarpium Citri Reticulatae distillation merges, be concentrated into relative density and be about 1.04～1.06 (60 ℃ of heat are surveyed), standing over night (more than 12 hours), get supernatant liquid filtering, in filtrate, add a certain amount of β-CD (be about full presciption medicine material total amount 2.5%), be concentrated into relative density and be 1.23～1.25 (60 ℃ of heat are surveyed), standby.Get above-mentioned alcohol extraction and water extracting liquid, mix, continuing to be concentrated into relative density in 50-70 ℃ is 1.30～1.32 (60 ℃ of heat are surveyed), vacuum drying (60 ℃ ,-0.08MPa), be ground into fine powder, standby.Other gets beta-schardinger dextrin-and places mortar in right amount, adds 4 times of water gagings and grinds evenly, and (beta-schardinger dextrin-: it is an amount of that ratio volatile oil) is got volatile oil in 8: 1, be made into 50% volatile oil ethanol liquid earlier, slowly join then in the mortar, ground 2 hours, in 50 ℃ of dryings, pulverize, standby.Get the Benexate Hydrochloride of above-mentioned dry extract and volatile oil, add an amount of sucrose, steviosin mixing,, cross 10～12 mesh sieves with 3% 30 POVIDONE K 30 BP/USP, 30 alcoholic solution system soft materials, 40 ℃ of dryings, granulate gets granule.

Embodiment 3

Get 14 parts of Herba Ardisiae Japonicaes, 7.1 parts in processed with honey Herba Ephedrae, 7.1 parts of Rhizoma Pinelliae Preparatum, 3 parts of Semen Armeniacae Amarums, 7.1 parts of Radix Peucedanis, 3 parts of Radix Arnebiae (Radix Lithospermi)s, according to the percolation under Chinese Pharmacopoeia version appendix IO fluid extract in 2000 and the extractum item, make solvent with 75% ethanol, flooded 24 hours, then in flow velocity (with the 1000g medical material) percolation of per minute 3～5ml, the collection liquid of filtering, add a certain amount of β-CD (be about full presciption medicine material total amount 5%), vacuum concentration is (below 60 ℃,-0.08MPa) be 1.10～1.25 (60 ℃ of heat are surveyed) to relative density, standby.Get 8 parts of Herba Schizonepetae, 3 parts of Pericarpium Citri Reticulataes, add 8 times of water gagings and soaked 3 hours, heating distillation extraction 4 hours is collected volatile oil and filtrate, and is standby.Get 3 parts of Radix Scutellariaes, 7.1 parts of Pumexes, 3 parts of Periostracum Cicadaes, 3 parts in Radix Glycyrrhizae mentions that with above-mentioned alcohol the medicinal residues after the distillation merge, the heating decocting is carried three times, amount of water is respectively 10 times of medical material amount, 6 times, 6 times, extraction time was respectively 1.5 hours, 1 hour, 1 hour, collect three times the water extract, filter and and Herba Schizonepetae, filtrate after the Pericarpium Citri Reticulatae distillation merges, be concentrated into relative density and be about 1.04～1.06 (60 ℃ of heat are surveyed), standing over night (more than 12 hours), get supernatant liquid filtering, in filtrate, add a certain amount of β-CD (be about full presciption medicine material total amount 2.5%), be concentrated into relative density and be 1.23～1.25 (60 ℃ of heat are surveyed), standby.Get above-mentioned alcohol extraction and water extracting liquid, mix, continuing to be concentrated into relative density in 60 ℃ is 1.30～1.32 (60 ℃ of heat are surveyed), vacuum drying (60 ℃ ,-0.08MPa), be ground into fine powder, standby.Other gets beta-schardinger dextrin-and places mortar in right amount, adds 4 times of water gagings and grinds evenly, and (beta-schardinger dextrin-: it is an amount of that ratio volatile oil) is got volatile oil in 8: 1, be made into 50% volatile oil ethanol liquid earlier, slowly join then in the mortar, ground 2 hours, in 50 ℃ of dryings, pulverize, standby.Get the Benexate Hydrochloride of above-mentioned dry extract and volatile oil, add an amount of sucrose, steviosin mixing, with 3% 30 POVIDONE K 30 BP/USP ₃₀Alcoholic solution system soft material is crossed 10～12 mesh sieves, 40 ℃ of dryings, and granulate gets granule.

Embodiment 4

Get 19 parts of Herba Ardisiae Japonicaes, 12 parts in processed with honey Herba Ephedrae, 12 parts of Rhizoma Pinelliae Preparatum, 7.9 parts of Semen Armeniacae Amarums, 12 parts of Radix Peucedanis, 7.9 parts of Radix Arnebiae (Radix Lithospermi)s, according to the percolation under Chinese Pharmacopoeia version appendix IO fluid extract in 2000 and the extractum item, make solvent with 75% ethanol, flooded 24 hours, then in flow velocity (with the 1000g medical material) percolation of per minute 3～5ml, the collection liquid of filtering, add a certain amount of β-CD (be about full presciption medicine material total amount 2.5%), vacuum concentration is (below 60 ℃,-0.08MPa) be 1.15～1.20 (60 ℃ of heat are surveyed) to relative density, standby.Get 8 parts of Herba Schizonepetae, 7.9 parts of Pericarpium Citri Reticulataes, add 8 times of water gagings and soaked 0.5 hour, heating distillation extraction 6 hours is collected volatile oil and filtrate, and is standby.Get 7.9 parts of Radix Scutellariaes, 12 parts of Pumexes, 7.9 parts of Periostracum Cicadaes, 7.9 parts in Radix Glycyrrhizae mentions that with above-mentioned alcohol the medicinal residues after the distillation merge, the heating decocting is carried three times, amount of water is respectively 8 times of medical material amount, 6 times, 6 times, extraction time was respectively 1.5 hours, 1 hour, 1 hour, collect three times the water extract, filter and and Herba Schizonepetae, filtrate after the Pericarpium Citri Reticulatae distillation merges, be concentrated into relative density and be about 1.04～1.06 (60 ℃ of heat are surveyed), standing over night (more than 12 hours), get supernatant liquid filtering, in filtrate, add a certain amount of β-CD (be about full presciption medicine material total amount 2.5%), be concentrated into relative density and be 1.23～1.25 (60 ℃ of heat are surveyed), standby.Get above-mentioned alcohol extraction and water extracting liquid, mix, continuing to be concentrated into relative density in 60 ℃ is 1.30～1.32 (60 ℃ of heat are surveyed), vacuum drying (60 ℃ ,-0.08MPa), be ground into fine powder, standby.Other gets beta-schardinger dextrin-and places mortar in right amount, adds 4 times of water gagings and grinds evenly, and (beta-schardinger dextrin-: it is an amount of that ratio volatile oil) is got volatile oil in 8: 1, be made into 50% volatile oil ethanol liquid earlier, slowly join then in the mortar, ground 2 hours, in 50 ℃ of dryings, pulverize, standby.Get the Benexate Hydrochloride of above-mentioned dry extract and volatile oil, add an amount of sucrose, steviosin mixing, with 3% 30 POVIDONE K 30 BP/USP ₃₀Alcoholic solution system soft material is crossed 10～12 mesh sieves, 40 ℃ of dryings, and granulate gets granule.

Embodiment 5

Get 18 parts of Herba Ardisiae Japonicaes, 9 parts in processed with honey Herba Ephedrae, 9 parts of Rhizoma Pinelliae Preparatum, 5.5 parts of Semen Armeniacae Amarums, 9 parts of Radix Peucedanis, 5.5 parts of Radix Arnebiae (Radix Lithospermi)s, according to the percolation under Chinese Pharmacopoeia version appendix IO fluid extract in 2000 and the extractum item, make solvent with 75% ethanol, flooded 24 hours, then in flow velocity (with the 1000g medical material) percolation of per minute 3～5ml, the collection liquid of filtering, add a certain amount of β-CD (be about full presciption medicine material total amount 2.5%), vacuum concentration is (below 60 ℃,-0.08MPa) be 1.15～1.20 (60 ℃ of heat are surveyed) to relative density, standby.Get 10.9 parts of Herba Schizonepetae, 5.5 parts of Pericarpium Citri Reticulataes, add 8 times of water gagings and soaked 0.5 hour, heating distillation extraction 6 hours is collected volatile oil and filtrate, and is standby.Get 5.5 parts of Radix Scutellariaes, 10.9 parts of Pumexes, 5.5 parts of Periostracum Cicadaes, 5.5 parts in Radix Glycyrrhizae mentions that with above-mentioned alcohol the medicinal residues after the distillation merge, the heating decocting is carried three times, amount of water is respectively 8 times of medical material amount, 6 times, 6 times, extraction time was respectively 1.5 hours, 1 hour, 1 hour, collect three times the water extract, filter and and Herba Schizonepetae, filtrate after the Pericarpium Citri Reticulatae distillation merges, be concentrated into relative density and be about 1.04～1.06 (60 ℃ of heat are surveyed), standing over night (more than 12 hours), get supernatant liquid filtering, in filtrate, add a certain amount of β-CD (be about full presciption medicine material total amount 2.5%), be concentrated into relative density and be 1.23～1.25 (60 ℃ of heat are surveyed), standby.Get above-mentioned alcohol extraction and water extracting liquid, mix, continuing to be concentrated into relative density in 60 ℃ is 1.30～1.32 (60 ℃ of heat are surveyed), vacuum drying (60 ℃ ,-0.08MPa), be ground into fine powder, standby.Other gets beta-schardinger dextrin-and places mortar in right amount, adds 4 times of water gagings and grinds evenly, and (beta-schardinger dextrin-: it is an amount of that ratio volatile oil) is got volatile oil in 8: 1, be made into 50% volatile oil ethanol liquid earlier, slowly join then in the mortar, ground 2 hours, in 50 ℃ of dryings, pulverize, standby.Get the Benexate Hydrochloride of above-mentioned dry extract and volatile oil, add an amount of sucrose, steviosin mixing, with 3% 30 POVIDONE K 30 BP/USP ₃₀Alcoholic solution system soft material is crossed 10～12 mesh sieves, 40 ℃ of dryings, and granulate gets granule.

Embodiment 6

The preparation process of the Benexate Hydrochloride of dry extract and volatile oil and embodiment 1 are identical.Be prepared into tablet only by the following method: the Benexate Hydrochloride of getting dry extract and volatile oil, the polyvinylpolypyrrolidone (PVPP) that adds fine powder weight 3 times of amounts microcrystalline Cellulose and 3%, mixing, 95% alcoholic solution system soft material with 2% 30 POVIDONE K 30 BP/USP 30 (PVP) pushed 16 eye mesh screens, 40～50 ℃ of dryings, granulate, the carboxymethyl starch sodium (CMS-Na) of adding 3% and 0.3% magnesium stearate mixing are pressed into 1000 (0.40g/ sheets), the bag film-coat promptly gets tablet.

Embodiment 7

The preparation process of the Benexate Hydrochloride of dry extract and volatile oil and embodiment 2 are identical.Be prepared into tablet only by the following method: the Benexate Hydrochloride of getting dry extract and volatile oil, the polyvinylpolypyrrolidone (PVPP) that adds fine powder weight 2 times of amounts microcrystalline Cellulose and 3%, mixing, 95% alcoholic solution system soft material with 2% 30 POVIDONE K 30 BP/USP 30 (PVP) pushed 16 eye mesh screens, 40～50 ℃ of dryings, granulate, the carboxymethyl starch sodium (CMS-Na) of adding 3% and 0.3% magnesium stearate mixing, compacting is (0.40g/ sheet) in flakes, the bag film-coat promptly gets tablet.

Embodiment 8

Step is identical with embodiment 1 with prescription, only uses sodium carboxymethyl cellulose (CMC-Na) to replace polyvinylpyrrolidone (PVP) _K-30, with alpha-cyclodextrin (α-CD) replace beta-schardinger dextrin-, with xylitol place of sucrose, steviosin.

Embodiment 9

Step is identical with embodiment 2 with prescription, only uses low-substituted hydroxypropyl cellulose (L-HPC) to replace polyvinylpyrrolidone (PVP) _K-30, replace beta-schardinger dextrin-with the N-LOK modified starch, with protein sugar place of sucrose, steviosin.

Embodiment 10

Step is identical with embodiment 5 with prescription, only uses Polyethylene Glycol (PEG) to replace polyvinylpyrrolidone (PVP) _K-30, with alpha-cyclodextrin (α-CD) replace beta-schardinger dextrin-, with high fructose place of sucrose, steviosin.

Pharmacodynamics test

Test material

1. medicine and reagent

1.1 be subjected to the reagent thing:

Chaste tree Fructus Pruni cough-stopping granule: it is a kind of product of the present invention, and its preparation method such as embodiment 5 are disclosed.Specification: the every 1ml of extractum contains the 4.25g crude drug.Clinical plan consumption 55g crude drug/day.

1.2 control drug

Cough due to wind and cold's electuary: main component has Pericarpium Citri Reticulatae, Rhizoma Zingiberis Recens, Rhizoma Pinelliae Preparatum, Pericarpium Citri Reticulatae Viride, Semen Armeniacae Amarum etc.Function warming lung for dispelling cold, expelling phlegm for arresting cough.Be used for affection of exogenous wind-cold, headache nasal obstruction, cough with copious phlegm, asthma uncomfortable in chest etc.Content: 5g (patent medicine)/bag, clinical consumption is 1 bag/time, 2 times/day, produce by Tai Ji group Sichuan Province Mianyang pharmaceutical factory, authentication code: ZZ-3282-defends in the river No. (1994) 010286, the accurate word of medicine, lot number: 0100105001.

Aspirin Enteric-coated Tablets: the long celebrating in Changchun Pharmaceutical (group) company limited product batch number: 20000104 specifications: 25mg/ sheet.

Peace is than first capsule: Hainan SANYE pharmaceutical factory lot number: 010822 specification: 2.5g/ grain.

Gentamycin 80,000 u/ml, lot number: north, 2001080 Hubei Province pharmaceutical factory.

Ampicillin 1g/ props up, lot number: 10012205 Huabei Pharmaceutic Co., Ltd.

1.3 test reagent:

Histamine phosphate: the beautiful pearl east wind in Shanghai Bioisystech Co., Ltd lot number: 020106.

Acecoline: Shanghai reagent three factory's lot numbers: 960508.

PertussisDiphtheriaTetanus triple vaccine: Hunan Hygienic ﹠ Epidemic Prevention Station provides lot number: 010719.

Cyclophosphamide: specification 20mg/ props up, Hualian Pharmaceutical Co., Ltd., Shanghai's lot number: 20010713.

Hydrocortisone: specification 25mg/ bottle, Shanghai Xinyi Pharmaceutical Factory lot number: 20001011.

Heparin sodium injection: specification 2ml/ props up, Wanbang Biochemically Pharmaceutical Co Ltd, Xuzhou's lot number: 0103103.

1.4 experiment Strain

Influenza virus A-prime Mus lung adapted strain FM1 is available from Inst. of Viruses, China Preventive Medicine Science Academy.

Influenza virus A 3 types, Coxsackie virus B 4 types (CoxB4) Parainfluenza type 1 viruss (HVJ), respiratory syncytial virus (RSV), adenovirus type III (Ad3) and rhinovirus 14 types.Respectively available from Virology Inst., Chinese Academy of Preventive Medical Science and institute of pediatrics.

1.5 test bacterial strain

Type strain: staphylococcus aureus (25923), Diplococcus pneumoniae (31002), Hemolytic streptococcus (32213), hemophilus influenza (58529), hemophilus influenza (58601) is not drawn mucositis bacterium (29103), and above bacterial strain is provided by Hunan Hygienic ﹠ Epidemic Prevention Station.

Clinical strain: staphylococcus aureus, Diplococcus pneumoniae, hemophilus influenza provide by microorganism teaching and research room of Hunan College of Traditional Chinese Medicine.

2. laboratory animal:

2.1 source and kind:

Kunming mouse, cleaning level, body weight 13～17g, male and female dual-purpose, the quality certification number: moving word: 20-011 number of Hunan doctor.

The Wistar rat meets the cleaning grade standard, body weight: 45～85g, and the male and female dual-purpose, the quality certification number: moving word: 20-022 number of Hunan doctor, above animal is provided by the laboratory animal department of the Chinese Academy of Sciences of Hunan Medical University.

Antiviral is tested used animal: Kunming mouse, body weight 14 ± 1g, male and female dual-purpose.The quality certification number is provided by laboratory animal room of Beijing Biological Product Inst.: the capital is moving is betrothed to (1999) No. 012.

3. test apparatus:

TP-1000 type electronic balance: Hunan instrument balance equipment factory, SE2020 type electronic balance: Shanghai Ao Hao company, L80-2 centrifugal precipitation mechanism: the Shanghai medical apparatus and instruments factory of making a leapleap forward, 721 type spectrophotometers: Shanghai analytical tool three factories, ultrasound atomizer, the S-888E type, Chinese-foreign joint Nanjing Dao Fen Electronics Co., Ltd. produces.

4. statistical method:

Measurement data with

Expression, the neat person of variance is with variance analysis and t check analysis in groups: the heterogeneity of variance person is with the rank test analysis.Enumeration data is with X ²Check analysis.

5. dosage setting:

The clinical children's's usage and dosage of chaste tree Fructus Pruni cough-stopping granule: warm boiled water, every day 3 times, each 4g, symphysis medicine 55g.Children's's body weight is calculated with 35kg, and children's's consumption every day is 1.57g crude drug/kg.Build coefficient conversion method by laboratory animal and people's clinical medicine dose is calculated, $d_B=d_A\×\frac{R_B}{R_A}\×\frac{\sqrt[3]{W_A}}{\sqrt[3]{W_B}}$ (R _Mice=0.06, R _Rat=0.09, R _Rabbit=0.093, R _{Cavia porcellus}=0.099R _{The people}=0.11).According to experimental result, with 0.5,1,2 times of amount of animal equivalent basic, normal, high dosage as large and small Mus, Cavia porcellus; With 1,2,4 times of amounts of animal equivalent basic, normal, high dosage as rabbit.Then the low dosage of laboratory animal, middle dosage, high dose are:

6. route of administration:

Gastric infusion, consistent with the clinical application approach of intending recommending.

Method and result

[primary symptom research]

One, to the influence of acute bronchitis rat model

(1) modeling experimental technique:

1. modeling and grouping:

Select 60 ♀ ♂ half and half of juvenile Wistar rats of body weight 55～85g, be divided into 6 groups at random, (1) normal group; (2) model group; (3), (4), (5) are the high, medium and low dosage group of chaste tree Fructus Pruni cough-stopping granule; (6) cough due to wind and cold's electuary group is put into the airtight glass container of 500ml with (2), (3), (4), (5), (6) group rat, and the burning sootiness of 50g Medicated cigarette end feeds O simultaneously ₂(contain 5%CO ₂), continue 1h, every day 1 time, continuous fume smoking 21 days, simultaneously nose drip Nai Shi mucositis bacterium (300,000,000/ml) 5,1 time weekly, totally 3 times.

2. medication:

With modeling administration simultaneously.Model group, normal control group gavage normal saline 0.5ml/ days respectively, continuous 21 days.

(2) detect index and result:

1. pathological examination

After the modeling 21 days, 1h after last 1 administration puts to death rat, and broken end is got blood, and to add anticoagulant heparin standby, is used for bactericidal assay and leukocyte trendization function test in the cell.After rat execution, immediately the trachea of each treated animal, bronchus, related lung tissue are fixed in 10% formalin solution in the lump, behind the 72h, take out trachea, bronchial tissue's piece, embedded section, HE dyeing is observed in microscopically.The results are shown in Table 1,2 and pathological examination report.

Table 1 chaste tree Fructus Pruni cough-stopping granule is to the reparation result of acute bronchitis rat model pathology damage

Through X ²Relatively; Compare with the normal control group ^{* *}P＜0.01; Compare △ △ P＜0.05 with model group

Table 2 chaste tree Fructus Pruni cough-stopping granule is to the integral result of acute bronchitis rat model pathology damage

Compare with the normal control group ^{* *}P＜0.01; Compare △ △ P＜0.05, △ △ △ P＜0.01 with model group

The result: by table 1, table 2 as seen, high, medium and low dosage group of chaste tree Fructus Pruni cough-stopping granule and model group relatively have significant difference, and prompting chaste tree Fructus Pruni cough-stopping granule has repair and alleviates its damaging action the pathology damage of acute bronchitis rat model.

2. bactericidal assay in the cell

Experimental technique

(1) rat that will test modeling is put to death, and broken end is got blood and added heparin anti-coagulating, isolates leukocyte Hank ' s liquid and is mixed with 5 * 10 ⁶/ ml cell suspension.

(2) opsonin: use 8ml Hank ' s liquid to add the fresh Mus serum of 2ml and be made into 20% fresh serum opsonin.

(3) antibacterial: staphylococcus aureus, microorganism teaching and research room in the court's keeps bacterial strain, and the depletion Staphylococcus aureus is mixed with 1 * 10 with it ⁸/ ml bacterial suspension.

(4) by following operation

(5) calculate:

The results are shown in Table 3.

Table 3 chaste tree Fructus Pruni cough-stopping granule bacteri ocidal test of leukocyte result

Compare with normal blank group: ^*P＞0.05; ^*P＜0.05; ^{* *}P＜0.01

Compare with model control group: △ P＞0.05; △ △ P＜0.05; △ △ △ P＜0.01

The result: by table 3 as seen, when 60min and 90min, the high, medium and low dosage of chaste tree Fructus Pruni cough-stopping granule relatively has significant difference with model control group, and prompting chaste tree Fructus Pruni cough-stopping granule can strengthen bactericidal action in the modeling rat cell.

3. leukocyte trendization function test (agarose Plating)

Method: rat is put to death after will testing modeling, and broken end is got blood and added anticoagulant heparin.Isolate leukocyte, be mixed with 2.5 * 10 with RPMI-1640 liquid ⁷/ ml cell suspension.Prepare the agarose plate by the agarose Plating, every plate is made a call to 24 holes, and pitch-row is 2.4mm, and a sample is measured in every row 3 holes.Interstitial hole adds leukocyte suspension 5 μ l, and outer hole adds escherichia coli 24h liquid culture filtrate 5 μ l, adds RPMI-1640 culture fluid 5 μ l with the hole and compares.After above-mentioned plate added a cover, put into warm box, put 37 ℃ of incubation 3h.Take out the every ware in back and add 3ml methanol 30min processing, continue to add 47% neutral formalin solution 3ml and fix, take out complete agar layer, the cell that is adhered fixed at the plate bottom dyes with Wright's stain, and washing is waited to do.Measure the distance that interstitial hole is divided a word with a hyphen at the end of a line to both sides with 400 times of biological microscope micrometers.The distance of dividing a word with a hyphen at the end of a line with the outside hole of interstitial hole cell (chemotactic factor) is A, and the distance of dividing a word with a hyphen at the end of a line to endoporus (culture fluid contrast) is B, is calculated as follows movement index, the results are shown in Table 4.

Table 4 chaste tree Fructus Pruni cough-stopping granule leukocyte trendization function test result

Compare with normal blank group, ^{* *}P＜0.01

Compare △ P＞0.05, △ △ P＜0.05, △ △ △ P＜0.01 with model group

The result: by table 4 as seen, chaste tree Fructus Pruni cough-relieving height, middle dosage group and model control group relatively have significant difference, and prompting chaste tree Fructus Pruni cough-stopping granule can increase the leukocyte movement index of modeling rat, improves leukocyte trendization function.

Two, antiinflammatory experiment

(1) influence of xylol induced mice auricle edema

Method: get 70 of body weight 13～17g male mice in kunming, be divided into 7 groups at random, normal blank group, model group, chaste tree Fructus Pruni cough-relieving height, in, low dose group, the aspirin group, cough due to wind and cold's electuary group, 10 every group, wherein normal blank group, model group gives distilled water, and the gastric infusion group is pressed table 5 gastric infusion respectively, every day 1 time, continuous 6d, behind time administration 1h, every mice left side ear front-back is not coated with the 0.05ml caused by dimethylbenzene xylene and swells, and auris dextra is not coated with to compare, behind the 30min, put to death animal, cut a left side, auris dextra is laid round auricle at same position respectively with 9mm diameter card punch, weigh with scale, as the swelling degree, data the results are shown in Table 5 through t check in groups with left and right sides ear difference.

The influence of table 5 chaste tree Fructus Pruni cough-stopping granule xylol induced mice auricle edema

Annotate: compare △ P＞0.05, △ △ P＜0.05, △ △ △ P＜0.01 with model control group; Compare with the blank group ^{* *}P＜0.01

The result: each dosage group of chaste tree Fructus Pruni cough-stopping granule and model group more all have significant difference, and prompting chaste tree Fructus Pruni cough-stopping granule xylol causes mice auricle swelling and has obvious inhibitory action.

(2) rat skin lower injection agar is caused the influence of granuloma hypertrophy.

Method: get 60 of body weight 55～85g male Wistar rats, be divided into 6 groups at random, 10 every group, under the etherization state, injection 2% agar solution under the Mus dorsal line, every 1ml presses administration every day of table 6 dosage or distilled water, after the administration 15 days, the jugular vein sacrificed by exsanguination is dissected, and separates the granuloma agar block, takes by weighing weight in wet base, data the results are shown in Table 6 through t check in groups.

Table 6 chaste tree Fructus Pruni cough-stopping granule causes the outgrowth influence of granuloma to rat skin lower injection agar

Annotate: compare with model control group: △ △ P＜0.05; △ △ △ P＜0.01

The result: high, medium and low dosage group of chaste tree Fructus Pruni cough-relieving and model control group relatively have significant difference, and prompting chaste tree Fructus Pruni cough-stopping granule has obvious inhibitory action to granuloma hypertrophy due to the subcutaneous injection agar.

Three, antitussive experiment

(1) strong aqua ammonia is caused the influence of mouse cough.

Method:, find out the minimum spray time (adopting the ultrasonic atomizatio method) that ammonia causes mice 100% cough and get 50 of body weight 13～17g Kunming mouses, ♀ ♂ half and half by prerun.Be divided into model control group at random by body weight and sex.The high, medium and low dosage group of chaste tree Fructus Pruni cough-relieving, cough due to wind and cold's electuary group, is pressed table 7 dosage gastric infusion or distilled water respectively by 10 every group, administration every day 1 time, continuous 3d, 1h after the last administration puts into mice the bell jar of volume 4L respectively, spraying into 27% ammonia 8 seconds stimulates mice to cause to cough (is the index of coughing to draw the abdomen, to face upward head, dehisce or cough), observe the cough number of times in mouse cough incubation period and the 3min, data the results are shown in Table 7 through t check in groups.

Table 7 chaste tree Fructus Pruni cough-stopping granule causes the influence of mouse cough to strong aqua ammonia

Annotate: compare with model control group: △ P＞0.05; △ △ P＜0.05; △ △ △ P＜0.01

The result: chaste tree Fructus Pruni cough-stopping granule height, in two dosage groups and matched group relatively, cough number of times and significant differences is all arranged incubation period, there were significant differences aspect the cough number of times for low dose group, and prompting chaste tree Fructus Pruni cough-stopping granule causes mouse cough to strong aqua ammonia and has obvious inhibitory action.

(2) to SO ₂The influence of induced mice cough

Method: get 50 of body weight 13～17g Kunming mouses.Be divided into model control group at random by body weight and sex, the high, medium and low dosage group of chaste tree Fructus Pruni cough-relieving, cough due to wind and cold's electuary group, 10 every group, matched group gavages distilled water, other each group is pressed table 8 dosage gastric infusion, every day 1 time, successive administration 3d, 1h after the last administration, mice is put into the wide mouthed bottle of 500ml, feed SO ₂6ml draws and coughs, and observes cough number of times in the cough latent period of record mice also and the 3min, and data are through t check in groups.The results are shown in Table 8.

Table 8 chaste tree Fructus Pruni cough-stopping granule is to the influence of SO2 induced mice cough

Annotate: compare with matched group: △ P＞0.05; △ △ P＜0.05; △ △ △ P＜0.01

The result: relatively cough number of times and significant difference is all arranged incubation period of chaste tree Fructus Pruni cough-stopping granule height, middle dosage group and matched group, prompting chaste tree Fructus Pruni cough-stopping granule is to SO ₂Cause and coughed obvious inhibitory action.

[inferior disease research]

One, the experiment of reducing phlegm

(1) to the phenol red influence of mice trachea excretion

Method: 1. get 60 of body weight 13～17g Kunming mouses, ♀ ♂ half and half, be divided into the blank group at random by body weight and sex, chaste tree Fructus Pruni cough-relieving height, in, low dose group, cough due to wind and cold's electuary group, every group 10, press table 9 dosage administration or distilled water respectively, successive administration 3 days behind the last administration 1h, is pressed the dosage of 0.1mg/10g, give the phenol red solution of every Mus ip0.5%, behind the 30min, put to death animal, peel off the trachea surrounding tissue, cut from thyroid cartilage to one section trachea of trachea bifurcation, put into the test tube that fills the 2ml normal saline, add 0.1ml1NaOH solution (1mol/L) again, survey the OD value in the 546nm place with 721 type spectrophotometers.

2. with the phenol red compound concentration of analytical pure the solution of 100 μ g/ml, 10 μ g/ml, 5 μ g/ml, 3 μ g/ml, 1 μ g/ml, 0.7 μ g/ml, 0.5 μ g/ml, 0.3 μ g/ml, 0.1 μ g/ml, measure its OD value, calculating its standard curve regression equation is y=0.1368+7.4673X (r=0.9988).

3. according to standard curve the OD value that records is converted into phenol red content, data the results are shown in Table 9 through t check in groups.

Table 9 chaste tree Fructus Pruni cough-stopping granule is to the phenol red influence of mice trachea excretion

Annotate: compare with model control group: △ △ P＜0.05; △ △ △ P＜0.01

The result: chaste tree Fructus Pruni cough-stopping granule height, middle dosage group and blank group relatively have significant difference, and prompting chaste tree Fructus Pruni cough-stopping granule can promote the phenol red effect of mice trachea excretion.

(2) to the influence of rat capillary glass-tube expectoration amount

Method: get 60 of body weight 55～85gWistar rats, ♀ ♂ half and half, be divided into the blank group at random by body weight and sex, chaste tree Fructus Pruni cough-relieving height, in, low dose group, cough due to wind and cold's electuary group, every group 10, water 10h is can't help in first fasting before the experiment, behind urethane ammonium 1g/kg intraperitoneal injection of anesthesia, it is fixing to face upward the position, cut off neck middle part skin and isolate trachea, on the thyroid cartilage lower edge between two cartilaginous rings with sharp-pointed entry needle Jianzha County one aperture, insert one of capillary glass tube then, make capillary tube just contact the trachea lower surface, make it to draw the sputum at trachea rear portion, the normal secretory volume of 2h before the record administration, after pressing table 10 dosage gastric infusion, observe the 2h secretory volume again.Data the results are shown in Table 10 through t check in groups.

Table 10 chaste tree Fructus Pruni cough-stopping granule is to the influence of rat capillary glass-tube expectoration amount

Annotate: compare with model control group: △ △ P＜0.05; △ △ △ P＜0.01

The result: three dosage groups of chaste tree Fructus Pruni cough-stopping granule and blank group are relatively, its difference has the significance meaning, and after the administration of high, medium and low three dosage groups average per hour secretory volume be before the administration more than 200%, illustrate that chaste tree Fructus Pruni cough-stopping granule has the effect of increase rat capillary glass-tube expectoration amount.

Two, the experiment of relievining asthma

Influence to histamine phosphate asthmatic model Cavia porcellus

Method: getting body weight is some of 120～180g Cavia porcelluss childhood, put into spray tank, spray into the equivalent mixed liquor of 2% acetylcholine and 0.4% histamine with ultrasound atomizer, each spray 10～15s, observe the asthma reaction of Cavia porcellus, select 50 of eligibles (asthma incubation period be no more than 150s be qualified), male and female are regardless of, be divided into 5 groups at random, be respectively 1. model blank group; 2. cough due to wind and cold's electuary group; 3.～5. the high, medium and low dosage group of chaste tree Fructus Pruni cough-relieving.After pressing randomly assigne each treated animal being pressed table 11 dosage gastric infusion 15min, repeat administration once again, behind the administration 30min, put into spray tank, similarity condition when selecting preliminary election for use sprays into the histamine phosphate mixed liquor, the asthma incubation period (have a convulsion from beginning to be sprayed to, the time till falling is the asthma incubation period of Cavia porcellus) of record Cavia porcellus.

Table 11 chaste tree Fructus Pruni cough-stopping granule causes the influence of asthmatic guinea pigs to histamine phosphate

Annotate: compare with matched group: ^*P＜0.05; ^*P＜0.01

The result: table 11 shows: in the cough-relieving of chaste tree Fructus Pruni, low dosage and blank group relatively, Cavia porcellus is lured breathe heavily significant prolongation incubation period, show that chaste tree Fructus Pruni cough-stopping granule can suppress the asthma that histamine phosphate causes Cavia porcellus.

[etiological study]

One, antiviral experiment

(1) material and reagent

Be subjected to the reagent thing: chaste tree Fructus Pruni cough-stopping granule, lot number: 200110901, provide by the Hunan Inst of Traditional Chinese Medicine.

Positive control drug: virazole: Hubei Province's institute of Pharmaceutical Industry is produced, and is crude drug, lot number: 950708, and mice is pressed clinical equivalent dosage (0.07g/kg/d) and irritates hello administration during experiment.

Cough due to wind and cold's electuary: Sichuan Province sheep pharmaceutical factory of Tai Ji group produces, lot number: 0100105001.Mice is pressed clinical equivalent dosage (2.10g/kg/d) and irritates hello administration during experiment.

Virus: influenza virus A-prime Mus lung adapted strain FM1, be used for the interior resisting virus experiment, available from Inst. of Viruses, China Preventive Medicine Science Academy ,-60 ℃ of cryogenic refrigerators are preserved standby.

Influenza virus A 3 types, Coxsackie virus B 4 types (CoxB4) Parainfluenza type 1 viruss (HVJ), respiratory syncytial virus (RSV), adenovirus type III (Ad3) and rhinovirus 14 types.Available from Virology Inst., Chinese Academy of Preventive Medical Science and institute of pediatrics, after going down to posterity, uses this laboratory respectively.

Cell: people's laryngeal carcinoma passage cell HEP-2 strain, available from Ministry of Public Health pharmaceutical biological product calibrating institute.After going down to posterity, uses this laboratory.

Cell culture fluid: Eagle ' the s MEM (a Japan day water Pharmaceutical Co., Ltd produces) that contains 10% calf serum, 0.29mg glutamine/ml, 100u/ml penicillin and streptomycin.

Cell maintenance medium: except that contained calf serum is 2%, all same culture fluid of other content.

Animal: Kunming mouse, body weight 14 ± 1g, male and female dual-purpose.By laboratory animal room of Beijing Biological Product Inst., the quality certification number: the capital is moving is betrothed to (1999) No. 012.

Key instrument: CO2 incubator, the loose Co., Ltd. of Japanese Yamato product.Inverted microscope, Japanese Olympus product.

(2) method and result

1. interior resisting virus experiment

(1) to the inhibitory action of mice influenza virus property pneumonia

Method: get 70 of mices, by body weight be divided into 7 groups at random, be respectively intact animal's matched group, viral infection matched group, three dosage groups of administration, virazole group, cough due to wind and cold's electuary group.Except that the normal control group, mice is slightly anaesthetized with ether, with 15 LD ₅₀Influenza virus drop nose infects, every 0.05ml.Feed administration from infecting to begin the previous day to irritate, every day 2 times, each 0.5ml, continuous 5 days, matched group under equal conditions distilled water filling was fed.Dissected after taking by weighing the mice body weight on the 6th day, the perusal pulmonary lesion, the degree of record pulmonary liver sample consolidation is won full lung and is weighed, and calculates the lung exponential quantity one by one, and obtains lung index suppression ratio.

Lung index=[heavy (the g)/body weight (g) of lung] * 100%

Statistical procedures is carried out in the t check between employing group as a result.See Table 12

Table 12 medicine is to the inhibitory action of mice influenza virus property pneumonia

Compare with the normal control group: ##P＜0.01

Compare with the virus control group: ^*P＜0.01, ^*P＜0.05

The result: table 12 result shows: be subjected to the height of reagent thing, the lung exponential quantity of middle dosage treated animal to be starkly lower than the virus control group, with the virus control group significant difference (P＜0.05) is arranged relatively, show that medicine all has the obvious suppression effect to the mice pneumonia that influenza virus causes when this dosage, suppression ratio is 18.08%.

(2) to the protective effect of influenza virus induced mice death

Method: get 120 of mices, be divided into 6 groups at random by body weight.Be respectively three dosage groups of chaste tree Fructus Pruni cough-stopping granule, virazole (0.07g/kg/d) group, cough due to wind and cold's electuary group (2.10g/kg/d) and viral infection matched group.Each organize mice with the slight anesthesia of ether after, with 2 LD ₅₀Influenza virus drop nose infects, every 0.05ml.Begin gastric infusion the previous day from infecting, every day 2 times, each 0.5ml, continuous 7 days.Accumulative total infects the death toll of back mice, calculates mortality rate, protective rate and increase in life span.The result adopts card side and T check to carry out statistical procedures.

Table 13 chaste tree Fructus Pruni cough-stopping granule causes the protective effect of dead mouse to influenza virus

Compare with the infection matched group: ^*P＜0.01, ^*P＜0.05

Table 14 chaste tree Fructus Pruni cough-stopping granule causes the protective effect of dead mouse to influenza virus

Compare with the infection matched group: ^*P＜0.01, ^*P＜0.05

The result: table 13,14 results show: in back 14 days of the mouse infection virus; the death toll of chaste tree Fructus Pruni cough-stopping granule 22.8,11.4g/kg/d dosage group mice infects matched group and obviously reduces; 11.4g/kg/d dosage group and matched group relatively have significant difference (P＜0.05), protective rate is 38.89%.The average survival natural law of mice infects matched group and obviously prolongs, and with matched group significant difference (P＜0.05) is arranged relatively, and rate elongation is 15% and 16.5%.Show that chaste tree Fructus Pruni cough-stopping granule causes dead mouse to influenza infection the certain protection effect is arranged when this dosage.

2. the extracorporeal antivirus effect of chaste tree Fructus Pruni cough-stopping granule experiment

(1) to the toxicity test of Hep-2 cultured cell

Method: medicinal liquid Eagle ' s culture fluid is done 1: 20-1: 1280 doubling dilutions, be added in the HEP-2 Tissue Culture Plate that grows up to monolayer, the 100ul/ hole, each dilution factor medicinal liquid is done 3 multiple holes, establishes the normal cell contrast simultaneously.Culture plate is put 37 ℃ of 5%CO ₂Cultivated in the incubator four days, observation of cell growing state under the every day inverted microscope determines that the minimum extension rate of obvious regression does not appear in cell, postpones during experiment and accomplishes minimum effective drug concentration (maximum dilution multiple).Calculate 50% toxic concentration (TC by the Reed-Muench method ₅₀) and maximal non-toxic concentration (TC ₀).

Table 15 chaste tree Fructus Pruni cough-stopping granule is to the TC of cultured cell ₀TC ₅₀(the mg crude drug/ml)

(2) to the influence of pathological changes caused by virus effect

Get the culture plate that grows up to cell monolayer, outwell culture fluid, inoculation 100TCID ₅₀Different virus liquid 50ul, put 37 ℃ of 5%CO ₂Absorption was outwelled viral liquid after 1 hour in the incubator, after keeping liquid and wash cell face 2 times with the Eagle ' s that does not contain calf serum, added corresponding dilution medicinal liquid 100ul/ hole.Establish virus control, positive control drug and normal cell contrast simultaneously.The final concentration of virazole adopts TC ₅₀1/4.Put 37 ℃ of 5%CO ₂Cultivate in the incubator, observation of cell pathological changes under the every day inverted microscope, when virus control group cell becomes ++ ++ the time record experimental result.Cytopathy is judged by six grade standards, and is calculated 50% valid density (EC by Reed-Muench ₅₀) and therapeutic index (TI).

One: the cell growth is normal, and no pathological changes occurs;

±: cytopathy is less than 10% of whole monolayer;

1: cytopathy accounts for below 25% of whole cell monolayer;

2: cytopathy accounts for below 50% of whole cell monolayer;

3: cytopathy accounts for below 75% of whole cell monolayer;

4: cytopathy accounts for more than 75% of whole cell monolayer;

Therapeutic index (TI)=TC ₅₀/ EC ₅₀

(3) result

Table 16 chaste tree Fructus Pruni granule is to the influence of pathological changes caused by virus

Annotate: "-" represents acellular pathological changes.

The result shows in the table: chaste tree Fructus Pruni cough-stopping granule when experiment in vitro, influenza virus, HVJ, RSV, CoxB ₄, Ad ₃Reach rhinoviral cytopathy and all alleviate to some extent, wherein HVJ, RSV, CoxB than the virus control group ₄And rhinovirus has significant difference (P＜0.05), EC ₅₀Be 0.4mg/ml and 3.4 with IT.Other viral not statistically significants.

Two, bacteriostatic experiment

(1) test material

1. bacterial strain

Type strain: staphylococcus aureus (25923), Diplococcus pneumoniae (31002), beta hemolytic streptococcus (32213), hemophilus influenza (58529), hemophilus influenza (58601) is not drawn mucositis bacterium (29103), and above bacterial strain is provided by Hunan Hygienic ﹠ Epidemic Prevention Station.

Clinical strain: staphylococcus aureus, Diplococcus pneumoniae, hemophilus influenza provides by microorganism teaching and research room of Hunan College of Traditional Chinese Medicine.

2. culture medium: Carnis Bovis seu Bubali cream soup agar culture medium; 5% Sanguis Leporis seu oryctolagi agar culture medium.

3. reagent: chaste tree Fructus Pruni cough-relieving extractum 4.25g crude drug/ml.Lot number: 200110901, the unit of providing: Hunan Inst of Traditional Chinese Medicine.

4. control drug:

Gentamycin 80,000 u/ml, lot number: 2001080, north, Hubei Province pharmaceutical factory.

Ampicillin 1g/ props up, lot number: 10012205, and Huabei Pharmaceutic Co., Ltd.

(2) method and result

1. bacteriostatic test in the body---to the protective effect of bacterial infection induced mice death

Method: with staphylococcus aureus, Diplococcus pneumoniae, the hemophilus influenza clinical strain is inoculated in the Carnis Bovis seu Bubali cream soup body culture medium, and put 37 ℃ of incubators and cultivated 8 hours, be 10 with the normal saline dilution ^-1, 10 ^-2, 10 ^-3, 10 ^-4The bacterium liquid of concentration is with the bacterium liquid difference lumbar injection infecting mouse of variable concentrations, every Mus 1ml, every group 10, observe the death condition of animal in 5 days, measure the MLD that test strain causes mice 90%-100% death, the MLD of bacterial strain is respectively: golden Portugal bacterium is 1 * 10 ⁸Cfu/ml, Diplococcus pneumoniae is 1.5 * 10 ⁸Cfu/ml, hemophilus influenza is 1.0 * 10 ⁹Cfu/ml.Get 360 of body weight 13～17g Kunming mouses, male and female half and half are divided into 18 groups at random, 20 every group, press table 17 dosage gastric infusion, matched group is given the isometric(al) normal saline, and administration is after 3 days, in the MLD1ml of the 4th day lumbar injection infecting mouse, continued administration again 3 days, observed and recorded infects the death toll of respectively organizing mice in one week of back, calculates mortality rate, passes through X with mortality rate ²Carry out statistical.

Table 17 chaste tree Fructus Pruni cough-stopping granule infects the protective effect of induced mice death to golden Portugal bacterium and Diplococcus pneumoniae

Annotate: compare △ △ P＜0.05 △ △ △ P＜0.01 with matched group

The result: result of the test shows, chaste tree Fructus Pruni cough-relieving high dose group infects the back mortality of mice to golden Portugal bacterium, hemophilus influenza clinical strain relatively has significant difference with matched group, in the cough-relieving of chaste tree Fructus Pruni, low dosage infects afterwards to the Diplococcus pneumoniae clinical strain that mortality of mice and matched group relatively have significant difference.

2. dull and stereotyped punch method

The plate (plain agar flat board/blood plate) of making even, with each the bacterial standard strain of inoculating loop picking, with method of scoring bacterium liquid evenly is coated with each media surface, cultivate 24h with the card punch punching (equidistant 4 holes/flat board) of diameter 6mm then, whether observe has inhibition zone to occur, by quantizing antibacterial circle diameter with this preliminary power of judging the trial drug bacteriostasis, result such as following table:

Table 18 chaste tree Fructus Pruni cough-stopping granule bacteriostatic test result

(golden Portugal bacterium, Diplococcus pneumoniae, group B streptococcus, do not draw the mucositis bacterium to make positive control drug, hemophilus influenza, hemophilus influenza make positive control drug) with gentamycin with ampicillin.

3. test tube method is surveyed the MIC and the MBC of medicine

By tube dilution method with culture medium with drug dilution become 1: 1,1: 2,1: 4,1: 8,1: 16,1: 32,1: 64,1: 128, every pipe 1ml adds 3 * 10 to every pipe ^-3The fresh bacterium liquid 0.1ml of cfu/ml dilution sets up the bacterial strain contrast simultaneously, the culture medium contrast.Staphylococcus aureus, Diplococcus pneumoniae, group B streptococcus are made positive control drug with ampicillin, and hemophilus influenza, hemophilus influenza make positive controls with gentamycin.Put 37 ℃ and cultivate 24h, observe the bacterial growth situation.Not show muddiness, the reagent concentration of integral asepsis growth is the minimum inhibitory concentration of medicine to this bacterial strain, selecting in the MIC test the effective inoculating loop of culture medium of asepsis growth gets-encircles and be inoculated on the agar culture medium that does not contain reagent, putting 37 ℃ of incubators cultivated 18 hours, observe the bacterial growth situation, the concentration of integral asepsis growth is the minimum bactericidal concentration (MBC) of medicine to this bacterial strain.

The results are shown in Table 19,20.

Bacteriostatic test (MIC) result of table 19 chaste tree Fructus Pruni cough-stopping granule

Bacteriostatic test (MBC) result of table 20 chaste tree Fructus Pruni cough-stopping granule

The result: result of the test shows, chaste tree Fructus Pruni cough-stopping granule to staphylococcus aureus, beta hemolytic streptococcus, Diplococcus pneumoniae, do not draw the mucositis bacterium that in various degree external bacteriostasis is all arranged, wherein to a little less than beta hemolytic streptococcus and the Diplococcus pneumoniae effect.

[support study dies]

One, immunologic function test

(1) to the influence of immunologic hypofunction mice carbon clearance

Method: get 60 of male mice in kunming childhood, be divided into 6 groups at random, the normal control group is irritated the stomach distilled water every day, other respectively organize every day SC hydrocortisone 50mg/kg once, cause immunologic hypofunction, and give the cough-relieving of chaste tree Fructus Pruni high, medium and low dosage respectively, cough due to wind and cold's electuary or distilled water, continuous 7 days, behind last administration 1h, every caudal vein injection india ink 10ml/kg is when 2min, 12min, get blood 20ul from the eyeground vein clump respectively, be added to 2ml0.1%Na ₂CO ₃Shake up in the solution, measure absorbance at the 680nm place in 721 type spectrophotometers, with the mice sacrificed by decapitation of having got behind the blood, its liver and spleen taken out weigh, index is cleaned up in calculating and index is cleaned up in correction.

Table 21 chaste tree Fructus Pruni cough-stopping granule is to the influence of immunologic hypofunction mice carbon clearance

Annotate: compare with the blank group: ^{* *}P＜0.01

Compare with model group: △ P＞0.05; △ △ P＜0.05; △ △ △ P＜0.01

The result: table 21 shows, high, medium and low dosage group of chaste tree Fructus Pruni cough-stopping granule and model control group relatively, its difference has the highly significant meaning, prompting chaste tree Fructus Pruni cough-stopping granule is to the be significantly improved effect of specific immunity of immunologic hypofunction mice due to the hydrocortisone.

(2) to the influence of immunologic hypofunction mice serum hemolysin

Method: get 60 of Kunming mouses childhood, ♀ ♂ half and half, be divided into 6 groups at random by body weight and sex, every group 10, blank group and model group are irritated the stomach distilled water, and all the other respectively organize the stomach of 0.2ml/10g filling by measure chaste tree Fructus Pruni cough-relieving height, in, low dosage, corresponding medicinal liquids such as cough due to wind and cold's electuary, continuous 7 days, except that the blank group, all the other respectively organize mice by the 40mg/kgSC cyclophosphamide for three days on end, cause the immunologic hypofunction model, every Mus lumbar injection 5% chicken erythrocyte suspension 0.2m1 on the 1st, behind the last administration 1h, get blood in the eyeground vein clump, separation of serum, with normal saline serum is diluted 100 times, get dilute serum 1ml, with 5% chicken erythrocyte suspension 0.5ml, 10% guinea pig serum 0.5ml mixes, incubation 30min in 37 ℃ of calorstats, put stopped reaction in 0 ℃ of refrigerator, centrifugal, get supernatant in 721 type spectrophotometer 540nm place colorimetrics, measure absorbance, other establishes the blank pipe zeroing that does not add mice serum, and data are through t check in groups.

Table 22 chaste tree Fructus Pruni cough-stopping granule is to the influence of immunologic hypofunction mice serum hemolysin

Compare with normal blank group ^{* *}P＜0.01; Compare △ P＞0.05 with model control group; △ △ P＜0.05; △ △ △ P＜0.01

The result: table 22 shows that each dosage group of chaste tree Fructus Pruni cough-stopping granule and model control group compare, and its difference has significant difference, and prompting chaste tree Fructus Pruni cough-stopping granule can significantly improve caused by cyclophosphamide immunologic hypofunction mice and induce the hemolysin ability.

Two, analgesic experiment

Three vaccines are caused the refrigeration function of fever in rabbits

Method: get some of body weight 1～1.5kg healthy rabbits, laboratory temperature was controlled at about 25 ℃, in preceding 3 days of test, survey normal anus temperature every day 2 times, choose surplus the rabbit 40 that body temperature change is not higher than 0.3 ℃, ear vein injection triple vaccine 2ml/kg body weight causes the hyperpyrexia pathological model.Survey body temperature behind the 1h, the person chooses 40 experimental rabbits and is divided into matched group at random as laboratory animal, the high, medium and low dosage group of chaste tree Fructus Pruni cough-relieving, cough due to wind and cold's electuary group more than 0.5 ℃ to raise.Give being subjected to the reagent thing to irritate stomach respectively, matched group ig distilled water wherein, 1h, 2h, 3h, 4h measure body temperature behind the medicine.The results are shown in Table 23.

Table 23 chaste tree Fructus Pruni cough-stopping granule is to the influence of fever in rabbits body temperature due to the triple vaccine

Annotate: compare with matched group: △ △ △ P＜0.01; △ △ P＜0.05

The result: each dosage group of chaste tree Fructus Pruni cough-stopping granule and matched group relatively have significant difference, and prompting chaste tree Fructus Pruni cough-stopping granule can suppress the rising of rabbit body temperature due to the triple vaccine, has clearing away heat and expelling pathogen in the exterior effect definitely.

Chaste tree Fructus Pruni cough-stopping granule shows the experimental studies results of acute bronchitis rat model: chaste tree Fructus Pruni cough-stopping granule has repair to the pathology damage of acute bronchitis rat model, and can strengthen bactericidal action in the modeling rat cell, can increase the leukocyte movement index of modeling rat, improve leukocyte trendization function.

By the antiinflammatory experiment as seen, chaste tree Fructus Pruni cough-stopping granule xylol induced mice auricle edema, rat skin lower injection agar granuloma is formed with stronger inhibitory action, has certain antiinflammatory action.

By the antitussive experiment as seen, chaste tree Fructus Pruni cough-stopping granule is to strong aqua ammonia and SO ₂Draw to cough and all can prolong cough latent period and reduce cough number of times in the 3min, have certain antitussive action.

By reducing phlegm experiment as seen, chaste tree Fructus Pruni cough-stopping granule can promote the phenol red excretion of mice trachea and increase rat capillary glass-tube expectoration amount, certain resolve phlegm effect is arranged.

By relievining asthma experiment as seen, chaste tree Fructus Pruni cough-stopping granule can make Cavia porcellus lure to breathe heavily significant prolongation incubation period, show that chaste tree Fructus Pruni cough-stopping granule can suppress the asthma that histamine phosphate causes Cavia porcellus.

The resisiting influenza virus experimental result shows in the chaste tree Fructus Pruni cough-stopping granule body: medicine has the obvious suppression effect 22.8, during 11.4g/kg/d dosage to the mice influenzal pneumonia; When 11.4g/kg/d dosage influenza virus being caused dead mouse has significant protective effect, and protective rate is 38.89%, and increase in life span is 16.5%%.

The experiment of chaste tree Fructus Pruni cough-stopping granule extracorporeal antivirus effect shows: chaste tree Fructus Pruni cough-stopping granule has certain inhibitory action to influenza virus, HVJ, RSV, CoxB4, Ad3 and rhinovirus.Chaste tree Fructus Pruni cough-stopping granule is when experiment in vitro, influenza virus, HVJ, RSV, CoxB4, Ad3 and rhinoviral cytopathy are all alleviated to some extent than the virus control group, wherein HVJ, RSV, CoxB4 and rhinovirus and virus control group are relatively, significant difference (P＜0.05) is arranged, and EC50 and TI are 0.4mg/ml and 3.4.

By bacteriostatic experiment as seen: chaste tree Fructus Pruni cough-stopping granule to staphylococcus aureus, Diplococcus pneumoniae, do not draw mucositis bacterium, Hemolytic streptococcus etc. that in various degree external bacteriostasis is arranged, golden Portugal bacterium, Diplococcus pneumoniae, the death of hemophilus influenza infecting mouse are had protective effect in various degree.Chaste tree Fructus Pruni cough-stopping granule has certain bacteriostasis.

By immunization experiment as seen, chaste tree Fructus Pruni cough-stopping granule can improve the carbon clearance index of immunologic hypofunction mice and induce the hemolysin ability, the improve effect of immunity of tool.

By analgesic experiment as seen, chaste tree Fructus Pruni cough-stopping granule can obviously suppress the rising of rabbit body temperature due to the triple vaccine, has certain refrigeration function.

In a word, by to research and antiinflammatory, the cough-relieving of acute bronchitis rat model influence, reduce phlegm, relieving asthma, antiviral, test such as antibiotic, immune, analgesic, the result shows: medicine provided by the invention has repair to the trachea pathology damage of acute bronchitis rat model, and can strengthen bactericidal action in the model rat cell, strengthen model rat leukocyte chemotactic function; Granuloma due to the xylol induced mice auricle edema, rat skin lower injection agar is formed with stronger inhibitory action, has certain antiinflammatory action; To strong aqua ammonia and SO ₂Draw and cough the equal number of times that can prolong cough latent period and reduce cough in the 3min, have certain antitussive action; Can promote the phenol red excretion amount of mice trachea and increase rat capillary glass-tube expectoration amount, have certain resolve phlegm effect; The energy inhibition has antiasthmatic effect preferably by the Cavia porcellus asthma that histamine phosphate causes; Medicine provided by the invention is external to parainfluenza virus I type (HVT), respiratory syncytial virus, Coxsackie virus B 4 type (CoxB ₄) and rhinovirus 14 types all demonstrate certain inhibitory action; In the inherent institute of the medicine body provided by the invention amount of reagent scope mice influenzal pneumonia is had obvious inhibitory action, middle dosage causes dead mouse to influenza virus significant protective effect; To staphylococcus aureus, streptococcus pneumoniae, do not draw mucositis bacterium, Hemolytic streptococcus etc. that certain external bacteriostasis is arranged, to golden Portugal bacterium, Diplococcus pneumoniae, the death of hemophilus influenza infecting mouse have protective effect in various degree, have certain bacteriostasis; Can improve the carbon clearance index of immunologic hypofunction mice and induce the hemolysin ability, the tool immunization that improves; Fever in rabbits due to the triple vaccine there is tangible refrigeration function.The result shows that medicine provided by the invention has the acute bronchitis of improvement symptom and antiinflammatory, cough-relieving, eliminates the phlegm, relievings asthma, antiviral, antibiotic, enhance immunity, refrigeration function.

Toxicological study

1. acute toxicity test

Children's chaste tree Fructus Pruni cough-stopping granule Cmax is 4.25g/ml, when amassing and give mice 3 times/24h with Cmax, maximum filling body of stomach, this moment, the cumulative maximum dosage reached 510.0g/Kg, be equivalent to 324.84 times of clinical consumption, animal does not see the overt toxicity effect, and the result shows that the disposable medication of this medicine is than safety.

2. long term toxicity test

Children's chaste tree Fructus Pruni cough-stopping granule 20g/kg, 40g/kg, three dosage continuous irrigations of 80g/kg stomach, 8 weeks of rat, this moment, basic, normal, high dosage was equivalent to 12.75,25.5,51 times of clinical dosage respectively, phase of administration as a result and restorative observation period (2 week) compare with the blank group, the general situation of rat, body weight gain, hematology and blood parameters, main organs coefficient, perusal and the equal no significant difference of mirror undertissue morphological observation or unusual show that it is safe using in the children's chaste tree Fructus Pruni cough-stopping granule clinical application scope.

Can't decide toxic traditional Chinese medicine material and incompatibility in the drug prescription provided by the invention, clinical preceding toxicological study proof this product drug safety is better.

It will be recognized by those skilled in the art, perhaps just can determine the equivalence replacement of a lot of specific embodiment of the invention described herein according to conventional experience.The disclosure content of the publication that the application quotes also all is incorporated herein with reform, with as a reference.Yet should be understood that scope of the present invention is not limited to the above-mentioned specific embodiment.Particularly point out at this, although only listed two kinds of preparations (granule and tablet) of medicine of the present invention in specific embodiment, it will be understood by those skilled in the art that the present invention is not limited thereto, other dosage forms of mentioning herein can be implemented fully.Therefore the present invention also can implement not according to these specific descriptions, and still within the scope of appended claim.

Claims (12)

1. preparation of drug combination method that is used for the treatment of children's exogenous cough, the ingredient of described compositions is made by the following weight proportion raw material:

5～15 parts of Herba Schizonepetae

10～20 parts of Herba Ardisiae Japonicaes

5～15 parts in processed with honey Herba Ephedrae

1～10 part of Semen Armeniacae Amarum

1～10 part of Radix Scutellariae

5～15 parts of Radix Peucedanis

5～15 parts of Rhizoma Pinelliae Preparatum

5～15 parts of Pumexes

1～10 part of Periostracum Cicadae

1～10 part of Pericarpium Citri Reticulatae

1～10 part of Radix Arnebiae (Radix Lithospermi)

1～10 part in Radix Glycyrrhizae

Described method comprises:

(1) takes by weighing Herba Ardisiae Japonicae, processed with honey Herba Ephedrae, Rhizoma Pinelliae Preparatum, Semen Armeniacae Amarum, Radix Peucedani, Radix Arnebiae (Radix Lithospermi) by weight, carry out percolation, collect the liquid of filtering, add inclusion agents, the vacuum concentration concentrated solution of winning;

(2) take by weighing Herba Schizonepetae, Pericarpium Citri Reticulatae by weight, use water distillation and extraction, collect volatile oil and filtrate;

(3) claim the medicinal residues of Radix Scutellariae, Pumex, Periostracum Cicadae, Radix Glycyrrhizae and step (1) and (2) gained to merge by weight, the heating decocting is carried, and collects the water extract, filters, Herba Schizonepetae, Pericarpium Citri Reticulatae extract the filtrate merging of back gained in filtrate and the step (2), concentrate, get second concentrated solution, leave standstill then, get supernatant liquid filtering, add inclusion agents in gained filtrate, reconcentration gets the 3rd concentrated solution;

(4) get described first concentrated solution and described the 3rd concentrated solution, mix, concentrate, drying is ground into fine powder;

(5) clathrate of forming by described volatile oil and inclusion agents by the proper proportion preparation;

(6) get the clathrate that fine powder that step (4) obtains and step (5) obtain, make suitable dosage form.

2. method according to claim 1, wherein, the ingredient of described compositions is made by the following weight proportion raw material:

8～13 parts of Herba Schizonepetae

15～22 parts of Herba Ardisiae Japonicaes

7～12 parts in processed with honey Herba Ephedrae

3～8 parts of Semen Armeniacae Amarums

3～8 parts of Radix Scutellariaes

7～12 parts of Radix Peucedanis

7～12 parts of Rhizoma Pinelliae Preparatum

7～14 parts of Pumexes

3～8 parts of Periostracum Cicadaes

3～8 parts of Pericarpium Citri Reticulataes

3～8 parts of Radix Arnebiae (Radix Lithospermi)s

3～8 parts in Radix Glycyrrhizae.

3. method according to claim 2, wherein, the ingredient of described compositions is made by the following weight proportion raw material:

10.9 parts of Herba Schizonepetae

18 parts of Herba Ardisiae Japonicaes

9 parts in processed with honey Herba Ephedrae

5.5 parts of Semen Armeniacae Amarums

5.5 parts of Radix Scutellariaes

9 parts of Radix Peucedanis

9 parts of Rhizoma Pinelliae Preparatum

10.9 parts of Pumexes

5.5 parts of Periostracum Cicadaes

5.5 parts of Pericarpium Citri Reticulataes

5.5 parts of Radix Arnebiae (Radix Lithospermi)s

5.5 parts in Radix Glycyrrhizae.

4. method according to claim 1, wherein, the suitable dosage form described in the step (6) is any one in tablet, pill, capsule, capsule tablet, powder, granule, suspending agent, gel, the paste.

5. method according to claim 4, wherein, described suitable dosage form is tablet or granule.

6. method according to claim 1, wherein, in described step (6), further add and be selected from diluent, stabilizing agent, plasticizer, binding agent, fluidizer, disintegrating agent, filler, lubricant, plasticiser, one or more adjuvants and/or additive in coloring agent, film former, flavour enhancer, sweeting agent, the antiseptic.

7. method according to claim 1, wherein said step (1) is carried out as follows:

Take by weighing Herba Ardisiae Japonicae, processed with honey Herba Ephedrae, Rhizoma Pinelliae Preparatum, Semen Armeniacae Amarum, Radix Peucedani, Radix Arnebiae (Radix Lithospermi) by weight, make solvent with 60～90% ethanol, flooded 16～36 hours, then based on the flow velocity percolation of 1000g medical material with per minute 2～7ml, the collection liquid of filtering, 1～5% the described inclusion agents that adds full presciption medicine material total amount, concentrate 40～75 ℃ and decompression lower part described first concentrated solution.

8. method according to claim 1, wherein said step (2) is carried out as follows:

Take by weighing Herba Schizonepetae, Pericarpium Citri Reticulatae by weight, steeped 0.5-3 hour with the water logging that 4-12 doubly measures, distillation extraction 4～10 hours is collected described volatile oil and filtrate.

9. method according to claim 1, wherein said step (3) is carried out as follows:

Claim the medicinal residues of Radix Scutellariae, Pumex, Periostracum Cicadae, Radix Glycyrrhizae and step (1) and (2) gained to merge by weight, add hot water that medical material amount 4-10 doubly measures and fry in shallow oil and carry 1-3 time, the time is 1-5 hour, collects the water extract, filter and with step (2) in filtrate behind the Herba Schizonepetae, Pericarpium Citri Reticulatae distillation merge, partial concentration obtains described second concentrated solution, leaves standstill then, filter, add an amount of beta-schardinger dextrin-in gained filtrate, reconcentration gets described the 3rd concentrated solution.

10. method according to claim 1, wherein said step (4) is carried out as follows:

Get described first concentrated solution and the 3rd concentrated solution, mix, concentrate at 50-70 ℃, drying is ground into fine powder.

11. method according to claim 1, wherein said step (5) is carried out as follows:

Earlier described inclusion agents is used The suitable solvent furnishing inclusion agents pastel, add the described volatile oil that obtains in the step (2) of suitable proportion then, mix homogeneously makes gained solution and described inclusion agents stick with paste the dress thing then and merges, grind the suitable time, final drying obtains volatile oil clathrate compound.

12. method according to claim 11, wherein, described volatile oil ethanol dilution.