CN113288954A

CN113288954A - Traditional Chinese medicine composition, preparation method and application thereof

Info

Publication number: CN113288954A
Application number: CN202110773728.8A
Authority: CN
Inventors: 田纪祥; 赵晓昂; 金日显; 张东
Original assignee: Institute of Materia Medica of CAMS
Current assignee: Institute of Materia Medica of CAMS
Priority date: 2021-07-08
Filing date: 2021-07-08
Publication date: 2021-08-24

Abstract

The invention discloses a traditional Chinese medicine composition, a preparation method and application thereof, belonging to the technical field of traditional Chinese medicine preparation, wherein the traditional Chinese medicine composition comprises the following components in parts by weight: 1-5 parts of coptis chinensis, 1-5 parts of cicada slough, 2-7 parts of wild buckwheat rhizome, 2-7 parts of folium isatidis, 6-14 parts of clam shell, 6-14 parts of talc and 1-4 parts of liquorice.

Description

Traditional Chinese medicine composition, preparation method and application thereof

Technical Field

The invention relates to the technical field of traditional Chinese medicine preparation, and in particular relates to a traditional Chinese medicine composition, a preparation method and application thereof.

Background

Pharyngitis is an inflammation of the pharyngeal mucosa, submucosal tissue, and is often part of an upper respiratory infection. According to the difference of the length of the disease course and the nature of pathological changes, the disease is divided into two major categories, namely acute pharyngitis and chronic pharyngitis. The acute pharyngitis can be further divided into 3 types of acute simple pharyngitis, acute edematous pharyngitis and acute necrotizing pharyngitis, and the acute simple pharyngitis is the most common. Chronic pharyngitis is also classified as type 3: chronic simple pharyngitis, chronic hyperplastic (or hypertrophic) pharyngitis, and chronic dry (or atrophic) pharyngitis. The pathological changes of the three have obvious differences. The attack is caused by factors such as climatic season factors, adjacent organ diseases, pathogenic microorganisms, adjacent organ diseases, physical or chemical stimulation and the like.

Wherein, cold can cause pharyngeal mucosa vasoconstriction, phagocyte number reduction, local resistance decline, dryness can influence pharyngeal mucus secretion and cilium peristalsis, reduce the cleaning and humidifying function to the air, directly cause irritation and damage to pharyngeal mucosa, weather change is large in winter and spring, indoor air circulation is poor, and resistance decline and pathogenic microorganism invasion are easily caused; acute and chronic inflammations of adjacent organs such as nasal cavity, nasal sinuses, oral cavity, teeth, gum, larynx, trachea, bronchus, etc. can cause pharyngitis by invading pharynx along mucosa, submucosal tissue, local lymph and blood circulation, or repeatedly stimulating pharynx by inflammatory secretion, or being forced to breath by respiratory obstruction of nasal disease; allergic constitution or diseases of the whole body such as rheumatic fever, gout, diabetes, heart disease, anemia, nephritis, tracheitis, emphysema, bronchiectasis, tuberculosis, liver cirrhosis, malnutrition and constipation caused by digestive system diseases can cause general resistance reduction and pharyngeal blood circulation disorder, and pharyngitis is further caused; including bacteria, viruses, spirochetes, rickettsiae and the like, are main pathogenic factors of acute pharyngitis, can be directly derived from air and diet, and can also be indirectly derived from blood circulation and lymphatic circulation; if the speech is too much, people like to eat spicy food, hot food, excessive smoking and drinking, and air pollution such as chemical gas and dust, the mucous epithelium and glands of the pharynx can be damaged. The common factors inducing pharyngitis are the damage to local defense system, over fatigue, mental stress, insufficient sleep and the like.

Typical symptoms of pharyngitis include: pharyngalgia (72%), pharyngalgia (70%), hoarseness (65%), dry and itchy throat (50%), fever (45%), and the like, the main symptoms of acute pharyngolaryngitis are acute onset, dry and burning throat at the beginning, and further pain, and the pharyngalgia is more obvious when swallowing saliva than when eating, and can be accompanied by fever, headache, inappetence and soreness of limbs, invasion to the throat, hoarseness and cough. The main symptoms of chronic pharyngolaryngitis are pharyngeal discomfort, dryness, itching and swelling, a large amount of secretion, burning pain, easy dryness and nausea, foreign body sensation, failure of swallowing and failure of swallowing. The above symptoms are particularly aggravated by a little more speech, after eating pungent food, fatigue or weather changes. Repeated attacks of acute pharyngitis can cause chronic pharyngitis, which also manifests as acute pharyngitis symptoms in cases of exacerbations. The pharyngitis treatment is except for avoiding the tobacco and the dust, treating the nasal and sinus diseases, recovering the nasal respiration, dissuading the patients from long talk, clearing throat, strengthening physique, giving more vitamin-rich food and other etiological treatments. The main treatment is local treatment, such as medicine buccal tablet and gargle, medicine coating for diminishing inflammation and astringing, ultrasonic atomization and inhalation, local sealing therapy of mandibular angle or pharyngeal arch and pharyngeal wall, physiotherapy, and hyperplastic lymphoid tissue treatment methods including laser, freezing, electrocoagulation, electrocautery or chemical medicine cauterization.

In recent years, due to the influence of factors such as air pollution, climate and environment, the incidence rate of pharyngitis is rising, and becomes the most common disease in throat diseases, and the incidence rate reaches more than 50 percent and has a rising trend. Pharyngitis can bring obvious local discomfort to patients, affect work and rest, and cause general symptoms and other diseases to cause more lesions. Therefore, the development of the pharyngitis drug has great social significance and market demand.

In the medicine treatment, most of the existing western medicines for treating acute and chronic pharyngitis and laryngitis are antibacterial and anti-inflammatory medicines, and the existing western medicines are easy to generate drug resistance after being taken for a long time and have unsatisfactory effects. Compared with western medicines for resisting inflammation and relieving pain, the traditional Chinese medicine has certain advantages for treating pharyngitis all the time and is more popular with people. Traditional Chinese medicine treatment mainly aims at dispelling wind and clearing heat, moistening lung and promoting phlegm, but most of the traditional Chinese medicine treatment only can temporarily play a role in relieving pharyngitis uncomfortable symptoms, the pharyngitis can still recur frequently after the traditional Chinese medicine treatment is stopped, the pharyngitis can not be effectively treated and cured, and different medicines are required to be replaced according to the types of the pharyngitis. Therefore, there is a need for a Chinese medicinal composition for treating and curing pharyngitis.

Disclosure of Invention

The invention aims to provide a traditional Chinese medicine composition for treating pharyngitis, which is used for solving the problems in the prior art.

In order to achieve the purpose, the invention provides the following scheme:

the invention provides a traditional Chinese medicine composition which comprises the following raw materials in parts by weight: 1-5 parts of coptis chinensis, 1-5 parts of cicada slough, 2-7 parts of wild buckwheat rhizome, 2-7 parts of folium isatidis, 6-14 parts of clam shell, 6-14 parts of talc and 1-4 parts of liquorice.

Further, the raw materials comprise the following components in parts by weight: 2-4 parts of coptis chinensis, 2-4 parts of cicada slough, 3-5 parts of wild buckwheat rhizome, 3-5 parts of folium isatidis, 7-9 parts of clam shell, 7-9 parts of talc and 2-3 parts of liquorice.

Further, the raw materials comprise the following components in parts by weight: 3 parts of coptis chinensis, 3 parts of cicada slough, 4 parts of wild buckwheat rhizome, 4 parts of folium isatidis, 8 parts of clam shell, 8 parts of talc and 2 parts of liquorice.

Further, the traditional Chinese medicine composition is prepared into pills, granules, tablets, capsules or oral liquid.

Further, the traditional Chinese medicine composition is prepared into tablets. Each tablet of berberine is calculated by berberine hydrochloride (C)₂₀H₁₈ClNO₄) Not less than 9.00mg, glycyrrhizic acid (C)₄₂H₆₂O₁₆) Not less than 1.50 mg.

Further, the traditional Chinese medicine composition also comprises physiologically acceptable auxiliary materials.

The etiology and pathogenesis of the pharyngolaryngitis are complex in clinic, and the treatment methods of the pharyngolaryngitis are different from the traditional Chinese medicine based on syndrome differentiation, such as wind-cold type, wind-heat type, external-cold and internal-heat type, external-wind and internal-heat type, yin deficiency and fire excess type, spleen and kidney yang deficiency type, lung and defensive intrinsic heat type and the like. In clinical treatment methods, more heavy injections are used for dispelling wind and dissipating heat, nourishing yin and moistening dryness, relieving sore throat and eliminating phlegm, and the treatment formulas for pharyngitis caused by stasis and heat dissipation are fewer. The invention discloses a traditional Chinese medicine composition for treating acute pharyngitis or chronic pharyngitis, which is prepared from seven traditional Chinese medicines, coptis chinensis is taken as a monarch drug, the traditional Chinese medicines are used for clearing away heat and promoting diuresis, wild buckwheat is used for clearing away heat and toxic materials, promoting blood circulation and removing dampness, folium isatidis is used for clearing away heat and toxic materials, cooling blood and stopping bleeding, cicada slough is used for dispelling wind and removing heat and relieving sore throat, clam shells and talc are used for clearing away heat and promoting diuresis, pathogenic factors have the effect of removing obstruction, and liquorice is used for relieving spasm and pain, moistening lung and relieving cough and harmonizing all drugs.

The invention also provides a preparation method of the traditional Chinese medicine composition, which comprises the following steps: extracting Coptidis rhizoma and folium Isatidis with ethanol for 2-3 times, each time with 5-7 times of ethanol for 1-1.5 hr, mixing extractive solutions, recovering ethanol, and concentrating under reduced pressure to relative density of 1.15-1.25 (measured at 60 deg.C) to obtain ethanol extract; extracting periostracum Cicadae, rhizoma Fagopyri Dibotryis, Concha Meretricis Seu Cyclinae, Glycyrrhrizae radix and pulvis Talci with water for 2-3 times, adding 7-9 times of water for each time, extracting for 1-1.5 hr for each time, concentrating the supernatant under reduced pressure to relative density of 1.15-1.20 (measured at 60 deg.C) to obtain water extract, mixing the above ethanol extract and water extract, and vacuum drying.

The invention also provides a preparation method for preparing the traditional Chinese medicine composition into tablets, which comprises the following steps: extracting Coptidis rhizoma and folium Isatidis with ethanol for 2-3 times, each time with 5-7 times of ethanol for 1-1.5 hr, mixing extractive solutions, recovering ethanol, and concentrating under reduced pressure to relative density of 1.15-1.25 to obtain ethanol extract; adding water into periostracum Cicadae, rhizoma Fagopyri Dibotryis, Concha Meretricis Seu Cyclinae, Glycyrrhrizae radix and pulvis Talci, extracting for 2-3 times, adding 7-9 times of water for each time, extracting for 1-1.5 hr, concentrating the supernatant under reduced pressure to relative density of 1.15-1.20 to obtain water extract, mixing the ethanol extract and water extract, vacuum drying to obtain dry extract, pulverizing the dry extract, adding adjuvants, granulating, drying, grading, tabletting, film coating, and packaging.

The invention also provides application of the traditional Chinese medicine composition in preparing a medicine for treating pharyngitis.

Further, the traditional Chinese medicine composition is used for preparing a medicine for treating acute pharyngitis or chronic pharyngitis caused by lung and stomach intrinsic heat.

The invention discloses the following technical effects:

the invention takes coptis as monarch drug. Coptis chinensis: bitter, cold and non-toxic. It is also indicated for heart, spleen, stomach, liver, gallbladder and large intestine meridians, and can clear heat and dry dampness, purge fire and remove toxicity. Wild buckwheat rhizome: has the functions of clearing away heat and toxic material, promoting blood circulation to cure carbuncle, expelling pus and removing blood stasis, and has the efficacy of clearing away heat and dispersing the heat-toxin in the lung and stomach which is attacking the throat swelling and pain, no matter whether pus is formed or not. It can be used both internally and externally. Folium isatidis: bitter and cold. Such as the liver, heart and stomach meridians. Has effects in clearing away heat, cooling blood, removing toxic substances, clearing away heat and toxic materials from upper jiao, and can be used for treating pharyngolaryngitis, tonsillitis, parotitis, common cold, fever, gingival hemorrhage, etc. The three medicines are compatible and synergistic, and have the effects of clearing heat and removing toxicity, cooling blood, removing blood stasis, relieving swelling and pain for the syndrome of intrinsic heat in lung and stomach. The cicada slough is salty, sweet and cool. It enters lung and liver meridians. Has effects in dispelling pathogenic wind, removing heat, relieving sore throat, promoting eruption, eliminating nebula, and relieving spasm. Can be used for treating wind-heat type common cold, pharyngalgia, hoarseness, measles without adequate eruption, rubella pruritus, conjunctival congestion, nebula, convulsion, and tetanus. Has the functions of dispelling wind-heat, relieving sore throat, ventilating lung and producing sound. The cicada slough is mainly used for dispelling wind-heat evil, expelling pathogenic qi, and clearing heat to the outside, and is combined with three heat-clearing and detoxifying drugs, so that wind can be dispersed and heat can be dissipated to the outside, cold and cool drugs can not be blocked, a pathologist 'diathermy changes qi' method can be achieved deeply, and a better curative effect is achieved on cough caused by throat itching. The traditional Chinese medicine composition disclosed by the invention can clear away heat and toxic materials, eliminate dampness and relieve sore throat, and reduce swelling and relieve cough, is suitable for acute pharyngolaryngitis or acute attack of chronic pharyngitis caused by intrinsic heat in lung and stomach, is in accordance with basic pathogenesis, and accords with basic treatment method of the disease.

A clamshell: salty and slightly cold, can clear heat and resolve phlegm; soften hardness and dissipate nodulation. The clamshell shell in the present invention serves to eliminate symptoms of pharyngeal discomfort caused by stagnation of phlegm-heat. Talc, raw licorice: the talcum powder is matched with the raw liquorice, can clear away heat and toxic materials, eliminate dampness and relieve sore throat, and reduce swelling and relieve cough, is suitable for acute pharyngolaryngitis or acute attack of chronic pharyngitis caused by heat accumulated in lung and stomach, is matched with basic pathogenesis, and accords with basic treatment method of the disease.

After the product is taken, the improvement rates of symptoms such as pharyngalgia, pharynx itch, dry pharynx, cough and the like are all about 80%, the improvement rates of mucosal congestion, edema and pharyngeal wall lymph follicular hyperplasia are all about 70%, no obvious adverse reaction is found after the product is taken, and the product has high clinical development value.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed to be used in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without inventive exercise.

FIG. 1 shows the TLC identification results of Coptidis rhizoma of the product prepared in example 4, wherein the reference group, preparation (sample) and berberine hydrochloride (reference substance) are sequentially arranged from left to right;

FIG. 2 is the thin layer identification of Glycyrrhiza uralensis of the product prepared in example 4, wherein the preparation (sample), glycyrrhetinic acid (control) and negative control group are shown from left to right;

fig. 3 is the result of the thin-layer identification of isatis leaf of the product prepared in example 4, wherein the preparation (sample), indigo (control 1), indirubin (control 2) and negative control group are sequentially arranged from left to right;

FIG. 4 shows the change of each cell in the distal portion of the pituitary gland in the model control group and the bolus group after three months;

FIG. 5 shows the change of each layer of cells of cerebral cortex and nerve cells of brain tissue in the model control group and the large dose group after three months.

Detailed Description

Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.

It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Further, for numerical ranges in this disclosure, it is understood that each intervening value, between the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.

It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The description and examples are intended to be illustrative only.

As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.

Cicada slough: the invention relates to a peel shell which falls off when nymphs of cicada insects such as Cryptotympana pustulata Fabricius emerge.

Coptis chinensis: the invention relates to a dried rhizome of Coptis chinensis francis Franch of Ranunculaceae.

Berberine in the coptis has the functions of antibiosis, antivirus and immunoregulation, and is consistent with the functional indications of the preparation, so the berberine is taken as one of the content detection indexes of the traditional Chinese medicine composition.

It has no cross-resistance with penicillin and streptomycin.

The present invention can reduce the number of pili on the surface of the thallus, so that the bacteria can not be attached to human cells to play a therapeutic role. It also has effects on helicobacter, and can relieve gastritis, gastric ulcer and duodenal ulcer.

Wild buckwheat rhizome: the invention relates to dried rhizome of Fagopyrum dibotrys (D.Don) Hara of Polygonaceae.

Folium isatidis: the present invention relates to a dried leaf of Isatis indigotica fort. belonging to the family Cruciferae.

The indigo and indirubin in the folium isatidis have the functions of resisting inflammation and relieving pain, and are consistent with the main functions of the product.

A clamshell: the invention relates to a shell of Cyclina sinensis Gmelin of the family Veneridae.

Chemical components: containing calcium carbonate, chitin, etc.

Talc: the present invention is a silicate mineral talc group talc, mainly containing magnesium silicate hydrate [ Mg ]₃(Si₄O₁₀)(OH)₂]. After the excavation, silt and miscellaneous stones are removed.

Chemical components: hydrous magnesium silicate.

Licorice root: the invention relates to dried roots and rhizomes of Glycyrrhiza uralensis Fisch.

Glycyrrhizic acid can be used for producing double salt or compound preparation with various alkaloids, antibiotics, amino acids, etc., and has effects of coordinating, solubilizing, increasing drug stability, improving bioavailability and reducing toxic and side effects. Many metal salts of glycyrrhizic acid can be properly absorbed by human body, and are not easy to cause accumulation and poisoning of elements. Glycyrrhizic acid and glycyrrhetinic acid in the liquorice have the functions of anti-inflammation, antibiosis, antivirus, antianaphylaxis and immunoregulation, and are consistent with the functional indications of the product, so the glycyrrhizic acid is taken as one of the content detection indexes of the product.

Example 1

The coptis and the indigowoad leaf are extracted by alcohol, and other five medicines such as cicada slough and the like are extracted by water: weighing 45g of coptis chinensis and 60g of folium isatidis according to the formula, adding 70% ethanol in an amount which is 6 times that of the coptis chinensis and the folium isatidis, extracting for 2 times, filtering ethanol liquid, and drying in vacuum. 45g of cicada slough, 60g of wild buckwheat rhizome, 120g of clam shell, 120g of talcum and 30g of liquorice, 8 times of water is added for extraction for 2 times, and the water extract is concentrated and dried in vacuum. Record as sample # 1.

Example 2

The coptis, the wild buckwheat and the indigowoad leaf are extracted by alcohol, and other four medicines such as cicada slough and the like are extracted by water: 30g of cicada slough, 80g of sea clam, 80g of talcum and 20g of liquorice, adding 8 times of water for extracting for 2 times, concentrating the extracting solution and drying in vacuum. 30g of coptis chinensis, 40g of wild buckwheat rhizome and 40g of folium isatidis, extracting with 6 times of 70% ethanol for 2 times, concentrating the ethanol solution, and drying in vacuum. Record as sample # 2.

Example 3

Seven medicines such as coptis are extracted with water: 30g of cicada slough, 80g of sea clam, 80g of talcum, 20g of liquorice, 30g of coptis chinensis, 40g of wild buckwheat rhizome and 40g of dyers woad leaf, adding 8 times of water to extract for 2 times, concentrating the extract and drying in vacuum. Record as 3# sample.

Examples 1-3 are the selection of extraction process route with pharmacodynamics and effective components of all the chemical components contained in the product as indexes, and the results are as follows:

the content of the index components berberine, glycyrrhizic acid and liquiritin is respectively measured by the samples, and the result shows that the index component content of the sample No. 1 and the index component content of the sample No. 2 are higher. The content of the No. 3 sample is low, and the analysis reason may be that in the decocting process, berberine and glycyrrhizic acid are combined into salt, which affects the dissolution of the effective components.

Comparison of drug effects and effective ingredients: the same dosage of different process extracts are used for testing mice, and the inhibition effect of the different process extracts of the pharyngitis prescription on the inflammatory granulation hyperplasia, namely whether the tested medicine has the anti-inflammatory and detumescent effects or not, is observed through a rat implantation cotton ball granuloma test, and the process route is selected. Experiments show that the positive drugs, the No. 1 and the No. 2 samples have obvious effects on the inhibition of the rat granuloma, and the water extract group has certain improvement on the indexes, but has no statistical significance; the sample No. 1 has more obvious effect and presents a certain dose-effect relationship, the result shows that the tested medicine has more obvious anti-inflammatory and detumescence effects, and the process route is more reasonable.

And (4) conclusion: by analyzing the comprehensive chemical components and pharmacodynamic experiment results, the 1# sample has higher content of index components including berberine, glycyrrhizic acid and liquiritin. Meanwhile, the pharmacodynamic result shows that the 1# sample has obvious pharmacodynamic result. Therefore, the process route was selected for sample # 1: ethanol extraction of the coptis chinensis and the folium isatidis, water extraction of the rest cicada slough and other medicines, and orthogonal optimization is carried out on the ethanol extraction process of the coptis chinensis and the folium isatidis and the water extraction process of the rest 5 medicines according to the results of the examples 1-3. The preferred optimal conditions are: extracting Coptidis rhizoma and folium Isatidis with 70% ethanol for 2 times, and adding 6 times of ethanol each time. The 5 kinds of herbs such as periostracum Cicadae, etc. preferably have the result of 2 times of water extraction, 8 times of water extraction each time, and 1 hour each time. The Chinese medicinal composition has the advantages that the daily dose of the Chinese medicinal composition is 32g, the yield of the extract is about 7 percent after extraction, and the daily dry extract dose is about 3g and basically meets the dose required by tablets. The water extract is directly dried without alcohol precipitation, which accords with the clinical medication characteristics and can fully ensure the drug effect. Meanwhile, the formula contains more mineral medicines, amino acid and other components, and calcium and magnesium ions in the mineral medicines such as talc, clam shells and the like are combined with effective components in the traditional Chinese medicine to form salts after the extracting solution is concentrated. In research, a large amount of effective components can be lost in the alcohol precipitation process. Therefore, the water extract of the method is not precipitated by alcohol. The selection of the drying method directly influences the content of the effective components, the currently common drying methods comprise vacuum drying and spray drying, and the individual components sensitive to heat can also be selected to be freeze-dried. In the concentration process, a large amount of effective components are precipitated in the concentration process due to the existence of calcium and magnesium ions, and a spray head is blocked in the spray drying process, so the spray drying is not selected in the method. In conclusion, the purification and drying process of the product selects the water extract to be concentrated and then directly dried in vacuum.

The product is taken about 2.4g of dry extract after extraction, and the extract is sticky, so the auxiliary materials are selected to have certain moisture-proof effect, good compressibility and economy. Through preliminary test, the auxiliary materials are calcium hydrophosphate and a small amount of magnesium stearate is added, so that the viscosity of the traditional Chinese medicine extract can be overcome, and the forming property and the compressibility are good. Selecting molding conditions: after the auxiliary materials are determined, the granulation and forming processes are studied. The proportion of the main drug and the auxiliary materials is determined, and the granules are prepared by 95 percent ethanol. To reduce the dose, auxiliary materials are supposed to be adopted: main drugs 1: 2, each tablet is prepared into 0.6g according to the conversion of the dose, 6 tablets need to be taken daily, and the dose is moderate. Example 4 is provided according to the above requirements.

Example 4

The present embodiment provides a Chinese medicinal composition, which comprises: 500g of cicada slough, 500g of coptis chinensis, 666.66g of wild buckwheat rhizome, 666.66g of dyers woad leaf, 1333.33g of clamshell, 1333.33g of talc and 333.33g of liquorice.

The preparation method of the traditional Chinese medicine composition comprises the following steps: the seven medicines are extracted twice by adding 70 percent ethanol into coptis and dyers woad leaf, each time is added with 6 times of ethanol, each time is extracted for 1 hour, the extracting solutions are combined, the ethanol is recovered, and the pressure reduction concentration is carried out until the relative density is 1.15-1.25 (measured at 60 ℃). Extracting 5 kinds of medicinal materials such as periostracum Cicadae with water for 2 times, 8 times of water for each time, 1 hr for each time, concentrating the supernatant under reduced pressure to relative density of 1.15-1.20 (measured at 60 deg.C), mixing with alcoholic extract such as Coptidis rhizoma, and vacuum drying. Pulverizing the dry extract, adding 170g of calcium hydrogen phosphate, 3g of magnesium stearate and 95% ethanol, granulating, drying, grading, tabletting, coating with film coat, making into 1000 tablets, and packaging. The product is a film tablet, and the content is brown yellow to brown granules.

Selection of coating conditions in this example: 3 batches of samples are prepared by selecting a coating material of Beijing Yingmao pharmaceutical Co Ltd according to the parameters provided by the Co, and the samples have the advantages of bright and uniform appearance and qualified disintegration. The coating conditions were as follows: the solid content of the coating liquid is 15 percent. Air inlet temperature: about 75 ℃, and the flow rate of the feed liquid is 150 mL/min. Solid content increasing: 2 to 3 percent.

In the product of the embodiment, the coptis chinensis is identified by thin-layer chromatography. The following identification methods were used: TLC identification of rhizoma Coptidis in the preparation. Berberine hydrochloride is used as reference substance. Grinding 5 tablets, adding 20ml of chloroform 0.2g, ultrasonically extracting for 10 minutes, filtering, and concentrating the filtrate to 1ml to obtain a test solution; taking berberine hydrochloride reference substance, adding chloroform to make into solution containing 0.5mg per 1ml as reference substance solution. According to a test of thin layer chromatography (appendix VI B of the first part of Chinese pharmacopoeia 2005), sucking 1 μ L of each of the two solutions, respectively dropping the two solutions on the same silica gel G thin layer plate, placing the solution in a developing tank pre-saturated with ammonia vapor by using benzene-ethyl acetate-isopropanol-methanol-ammonia water (6: 3: 1.5: 1.5: 0.5) as a developing agent, developing, taking out, drying in the air, and inspecting under an ultraviolet lamp (365 nm). In the chromatogram of the test solution, the same yellow fluorescent spot appears at the position corresponding to the chromatogram of the reference solution, the negative reference solution has no interference, the result is shown in figure 1, and the result shows that the spot in the thin-layer chromatogram is clear, the separation effect is good, the reproducibility is good, and the specificity is strong.

Folium Isatidis thin layer chromatography identification in the product. The identification method comprises the following steps: glycyrrhetinic acid was used as a control. Grinding 5 tablets of the product, adding 2ml of hydrochloric acid and 25ml of chloroform into 1.2g of the ground product; heating and refluxing for extraction for 1 hour, filtering, and concentrating the filtrate to 2ml to obtain a sample solution; glycyrrhetinic acid control solution is prepared by adding chloroform to obtain solution containing 0.5mg per 1 ml. Performing thin layer chromatography (appendix VI B of the first part of Chinese pharmacopoeia 2005), sucking 1 μ L of control solution and 2 μ L of sample solution, respectively dropping on the same silica gel G thin layer plate, developing with petroleum ether (60-90 deg.C) -benzene-ethyl acetate-glacial acetic acid (10: 20: 7: 0.5) as developing agent, taking out, air drying, spraying with phosphomolybdic acid ethanol solution, and heating at 105 deg.C until the spots are clearly developed. The spot with the same color appears on the corresponding position of the chromatogram of the test solution and the chromatogram of the reference solution, the negative reference solution has no interference, the result is shown in figure 2, and the figure 2 shows that the spot in the thin-layer chromatogram is clear, the separation effect is good, the reproducibility is good, and the specificity is strong. The thin-layer identification reference substance of the liquorice in the Chinese pharmacopoeia is glycyrrhizic acid, the thin-layer identification of the glycyrrhizic acid is selected at first in the experiment, but the interference of other components in the formula is large, the glycyrrhizic acid cannot be identified, and the liquorice is finally determined to be hydrolyzed into the glycyrrhetinic acid through a plurality of experiments, the polarity of the glycyrrhetinic acid is small, the interference is avoided, and the thin-layer identification reference substance of the liquorice in the formula can be used.

The liquorice in the product is identified by thin-layer chromatography. The identification method comprises the following steps: indigo and indirubin were used as control. Taking 5 tablets of the product, grinding, taking 1.5g, adding 30ml of chloroform, carrying out ultrasonic extraction for 20 minutes, filtering, and concentrating the filtrate to 1ml to be used as a test solution. And additionally, adding chlorine into the indirubin and the indigo reference substance to imitate a reference substance saturated solution as a reference substance solution. Performing thin layer chromatography (appendix VI B of the first part of Chinese pharmacopoeia 2005) by sucking 2 μ L of indirubin control solution, 10 μ L of indigo control solution and 3 μ L of sample solution, respectively dropping on the same silica gel G thin layer plate, developing with benzene-chloroform-acetone (5: 4: 1) as developing agent, taking out, and air drying. In the chromatogram of the test solution, the same blue spot and light purple spot appear at the corresponding positions of the chromatogram of the reference solution, and the negative reference solution has no interference. The result is shown in figure 3, and the result in figure 3 shows that the spots in the thin-layer chromatography are clear, the separation effect is good, the reproducibility is good, and the specificity is strong. In the Chinese pharmacopoeia, thin-layer identification reference substances of indigotin and indirubin are used as comparison products, the concentration of the thin-layer identification reference substances is 1mg/ml, but in the actual operation, the obtained indigotin and indirubin reference substances are found to have solubility which is not too good and is less than 1mg/ml, and through repeated grope, the saturated solution of the indigotin and the indirubin is finally determined to be used as the reference substance solution.

The detection is carried out according to the tablet inspection item in the first appendix of edition 2005 of Chinese pharmacopoeia, and the content difference, disintegration and microbial limit inspection of 3 batches of samples all accord with the regulations.

Measuring berberine hydrochloride and glycyrrhizic acid content by high performance liquid chromatography, selecting extraction conditions, performing system applicability test, examining linear relationship, stability, precision, repeatability, recovery rate, etc., establishing content measuring method for berberine hydrochloride and glycyrrhizic acid, and determining content limit of berberine hydrochloride and glycyrrhizic acid according to content measuring results of 3 batches of samples, i.e. each tablet of the product contains berberine calculated by berberine hydrochloride (C)₂₀H₁₈ClNO₄) Not less than 9.00mg, and glycyrrhizic acid (C)₄₂H₆₂O₁₆) Not less than 1.50 mg.

The result shows that the method has accurate content measurement and good repeatability.

The stability test of the product: and (3) packaging by adopting a polyethylene bottle, and inspecting the change condition of each index of the sample under the conditions of an accelerated test and a normal-temperature test. The sample of the example 4 is measured three times, and is subjected to an accelerated test for 6 months and a long-term sample retention test for 6 months, and the characteristics, the inspection, the identification, the content measurement and the microbial limit of the sample are subjected to investigation tests, and the test results show that the product has no obvious change in each index and good stability when being examined under the test conditions. According to the accelerated investigation result and the room temperature sample retention investigation result, the effective period of the traditional Chinese medicine composition is preliminarily determined to be 1.5 years.

Through process verification, quality research and stability investigation of 3 batches of samples, the production process is reasonable, the quality is controllable, and the properties are stable.

Pharmacological toxicology (taking the sample prepared in example 4 as an example)

1.1 major pharmacodynamic Studies

1.1.1 Effect of this product on animal model of acute pharyngitis in rats (Ammonia water spray)

Test materials:

(1) animal(s) production

Wiset rats, male, 190-220 g, purchased from Beijing Wintolite laboratory animal technologies, Inc. License number: SCXK (Jing) 2007 + 0001; the animal is bred in the animal center (in a barrier environment) tested by the institute of traditional Chinese medicine of the Chinese academy of traditional Chinese medicine, and the license number is as follows: SYXK (Kd) 2005-0028; qing II mouse material, license number: SCXK (Jing) 2005-.

(2) Drugs and agents

The tested drugs are: example 4 a product was prepared.

Honeysuckle and scutellaria granules, batch number: 090301, it is produced by Jiangxi Jimin trusted pharmaceutical Co.

Reagent: ammonia water: the Beijing chemical plant with the concentration of 25 percent to 28 percent is produced, and the batch number is as follows: 20081016.

formaldehyde solution: chemical reagent of national drug group, batch number: 20070905.

(3) apparatus and instrument

Type of throat sprayer: l56- -03, manufactured by Jiading district medical facilities, Shanghai City. Shanghai food and medicine monitor 2005 (Standard) No. 1541353.

Type of precision analytical balance: 13P110S

Blood cell counter model: MEK-6318K manufactured by Japan optoelectronics industry zhu-ji.

Pathology-related apparatus: full-automatic hydroextractor: shandon Excelsior ES, uk; a paraffin embedding machine: shandon histoentre 3, uk; rotating type slicing machine: shandon Finesse 325, uk; full-automatic dyeing machine: shandon variostatin Gemini, uk; microscope and image analysis system: olympus BX51, japan.

The method comprises the following steps:

(1) dose setting:

the tested drugs are: a human. The dosage is as follows: 32g crude drug/60 Kg body weight/day, i.e. 0.037g ointment/Kg body weight/day. The large dose is 0.444g extract/Kg (about 12 times of the clinical dosage), the medium dose is 0.222g extract/Kg (about 6 times of the clinical dosage), and the small dose is 0.111g extract/Kg (about 3 times of the clinical dosage).

Positive drugs: the dose of Yinhuang granules 0.8g/kg is 6 times of the clinical dose of human.

(2) Route of administration and volume:

all the drugs are orally administrated by gavage, 10ml/kg body weight, and distilled water with the same volume is given to a blank control group and a model control group.

(3) Test method

Grouping administration: the 58 Wiset rats are randomly divided into a blank control group, a model control group, a positive medicine group and a traditional Chinese medicine composition high, medium and low dose group, and each group except the blank control group comprises 10 animals. The drug administration is started on the day of modeling, and the traditional Chinese medicine composition is administered in a high, medium and low dose group and a honeysuckle and yellow granule group by intragastric administration once/day. The administration was carried out for a total of six days.

Molding: the animals except the blank control group were divided into groups and sprayed with 4.2% ammonia water for 1 time, and 3 pressure per time. Spraying was continued for 3 days. The blank control group was sprayed with drinking water to the pharynx by the same method as each model group.

Animals in group 7d were bled retroorbitally for hemogram determination, and sacrificed immediately to remove pharyngeal mucosa (mounted on cardboard with a pin) for pathological sectioning.

Note: the death of 2 animals resulted from the molding process (one each for model control and positive drug)

And (3) data statistics:

the hemogram of each animal is expressed by the mean coefficient plus or minus standard deviation, the tested drug group and the positive control group are compared with the model control group, and the result is statistically analyzed by a single variance of a sps software.

Semi-quantitative counting method: pathological changes are classified into commonality changes and subtotal changes (i.e., deep ulceration, focal necrosis in the deep layer of the posterior pharyngeal wall, lymphadenectasis, and prominence of the pharyngeal wall, myxoglandular cyst formation.) and one change per animal, one + for 1, 2 for + for 2, 3 for + for 3, 4 for + for 4, and 5 for + for 5.

The results are shown in tables 1-3.

Table 1: influence of traditional Chinese medicine composition on hemogram of rat acute pharyngitis model

The results show that: the product has no obvious influence on hemogram of model animal.

Pathological observation results:

the rats (8) in the placebo group showed no detrimental changes in the posterior pharyngeal wall structure. After three months, the cells on the distal part of the pituitary gland of the model control group and the large dose group are not abnormal (figure 4); after three months, nerve cells at all levels of brain tissues of the model control group are not degenerated, the structure is normal, cells at each layer of cerebral cortex of the large-dose group are not degenerated, and interstitial substance is not infiltrated (figure 5); three sections of the rear pharyngeal wall, namely the nasopharynx, the oropharynx and the laryngopharynx, all show normal structures. The nasopharynx and the laryngopharynx are covered with pseudostratified ciliated columnar epithelium, and the oropharynx is covered with non-cornified stratified squamous epithelium. A certain number of mucous glands are distributed in the inherent layer, and the submucosa is thinner and is not clear with the boundary of the inherent layer.

Model group rats (9) showed significant lesion changes in the posterior pharyngeal wall structure. The common change is two-fold, namely that all rats find a plurality of superficial ulcers (necrosis and abscission of the epithelium 1/3) on the mucous epithelium of the pharyngeal posterior wall, the tissues around the ulcers swell, blood vessels of the lamina propria and the submucosa expand, congestion and inflammatory cell infiltration are caused, and the distance between the epithelium and the muscular layer is increased. Secondly, all rats find that the oral pharynx proportion of the epithelium on the posterior pharyngeal wall is increased and certain degree of cornification phenomenon appears. Third, all rats found hyperplasia of mucous glands under the epithelium of the posterior pharyngeal wall, and the hyperplasia of mucous glands could extend to the muscular layer. The specific changes include: deep ulceration, focal necrosis in the deep layer of the posterior pharyngeal wall, enlargement of lymph follicles and protrusion of the pharyngeal wall, and formation of mucous gland cysts.

The positive drug control group rats (9) showed no significant damaging changes in pharyngeal posterior wall structure. No obvious pathological changes are seen in the three sections of nasopharynx, oropharynx and laryngopharynx of the posterior pharyngeal wall. The nasopharynx and the laryngopharynx were covered with pseudostratified ciliated columnar epithelia, and 4 animals had a superficial ulcer of mucosal epithelium; the oropharynx of most animals is covered with a keratinized stratified squamous epithelium. The mucous glands in the lamina propria extend to the muscular layer in many places. One lymph follicular enlargement and protruding pharyngeal wall appear in 1 animal, and focal necrosis in the deep layer of the pharyngeal posterior wall appears in 2 animals, and pathological changes such as deep ulcer and mucocele formation do not appear.

The posterior pharyngeal wall structures of the rats (10) in the high dose group showed some lesion changes, but lesion changes were significantly reduced compared to the rats in the model group; compared with the positive drug rats, the pathological changes of the two groups are similar. No obvious pathological changes are seen in the three sections of nasopharynx, oropharynx and laryngopharynx of the posterior pharyngeal wall. The nasopharynx and the laryngopharynx were covered with pseudostratified ciliated columnar epithelia, and 4 animals had a superficial ulcer of mucosal epithelium; the oropharynx of most animals is covered with a keratinized stratified squamous epithelium. The mucous glands in the lamina propria extend to the muscular layer in many places. 1 animal has deep ulcer without pathological changes such as deep focal necrosis of pharyngeal posterior wall, lymphangioleiomyomatosis, pharyngeal wall protrusion, and mucinous cyst formation.

The pharyngeal posterior wall structure of the medium-dose group rats (10) showed slight damaging changes, which were significantly reduced compared to the model group rats; compared with the positive drug rats, the pathological changes of the medium dose group are heavier. Three segments of the posterior pharyngeal wall, the nasopharynx, the oropharynx and the laryngopharynx all showed slight pathological changes. The nasopharynx and the laryngopharynx are covered with pseudostratified ciliated columnar epithelia, and superficial ulcer of mucosal epithelium at 1 part (or 2 parts) appears in 6 animals; the oropharynx of 9 animals was covered with keratinized stratified squamous epithelium. The mucous glands in the lamina propria extend to the muscular layer in many places. The ulcer of the posterior pharyngeal wall of 2 animals is deepened, the ulcer of the posterior pharyngeal wall of three sections of 1 animal is found, and the pathological changes of focal necrosis of the deep posterior pharyngeal wall, enlargement of lymph follicles, protrusion of the pharyngeal wall, formation of mucous gland cyst and the like are not seen.

The pharyngeal posterior wall structure of the small-dose group of rats (10) shows more obvious damage change, and compared with the model group of rats, the damage change is reduced to a certain extent; compared with positive rats, the pathological changes of the small-dose group are obviously heavier. Three sections of the rear pharyngeal wall, namely the nasopharynx, the oropharynx and the laryngopharynx, show obvious pathological changes. The nasopharynx part and the laryngopharynx part are covered with pseudo-stratified ciliated columnar epithelia, and superficial ulcer of mucosal epithelium at 2 places (or more than 2 places) appears in 7 animals; the oropharynx of 9 animals was covered with keratinized stratified squamous epithelium. The mucous glands in the lamina propria extend to the muscular layer in many places. The pathological changes of deep ulcer of 4 animals, ulcer found in three sections of the posterior pharyngeal wall of 2 animals, focal necrosis of deep layer of the posterior pharyngeal wall of 1 animal, lymphadenectasis and protrusion of pharyngeal wall of 3 animals, mucocele formation of 1 animal and the like.

TABLE 2 evaluation chart of pathological changes of pharyngeal backwall of model animal

The results show that: the degree of injury of tissues of the posterior pharyngeal wall is counted and scored according to the commonalities, deep ulcer, focal necrosis, lymphadenectasis and colistinosis. The results show that the tissue of the pharyngeal posterior wall of the model rat is seriously damaged compared with the blank group; compared with a model group, the injury of the pharyngeal posterior wall of each dose group of the product is obviously relieved, which shows that the product has a protective effect on the pharyngeal posterior wall tissue injury of an acute pharyngitis animal model.

Table 3 shows the effect of the composition on pathological changes of pharyngeal backwall tissue of rats with acute pharyngitis

The ammonia water stimulates the acute pharyngitis animal model is after stimulating the pharyngeal mucosa through the local spray of high concentration, pharyngeal congestion and swelling, form acute inflammation, its morbidity, pathological manifestation and pharyngitis are similar clinically, there are many superficial ulcers in pharyngeal wall mucosa epithelium after the model is made, the tissue swelling around the ulcer, inherent layer and submucosal layer blood vessel expansion congestion, inflammatory cell infiltration, the distance between epithelium and muscular layer increases, pharyngeal wall epithelium oropharyngeal portion proportion increases, appear certain degree of cornification phenomenon and pharyngeal wall epithelium mucus gland hyperplasia simultaneously behind the pharynx, the mucus gland of hyperplasia can expand to the muscular layer. By observing the effect of the product on acute inflammation, the product can obviously relieve local damage of the pharyngeal posterior wall and has obvious protective effect on congestion, swelling, ulcer and necrosis of pharyngeal mucosa.

1.1.2 Effect of this product on anti-inflammatory and detumescence effects of rats (Cotton ball granuloma test)

Test materials:

(1) animal(s) production

SD rats, male, 120-140 g, purchased from Beijing Wittiulihua laboratory animal technologies, Inc. License number: SCXK (Jing) 2007 + 0001; the animal is bred in the animal center (in a barrier environment) tested by the institute of traditional Chinese medicine of the Chinese academy of traditional Chinese medicine, and the license number is as follows: SYXK (Kd) 2005-0028. Qing II mouse material, license number: SCXK (Jing) 2005-.

(2) Drugs and agents

The tested drugs are:

example 4 the resulting extract was prepared.

Aspirin enteric-coated tablets (white tablets), size: 25 mg/tablet, manufactured by Beijing Pacific pharmaceutical Co., Ltd., lot number: 070901.

ether, purchased from national pharmaceutical group chemical agents limited, lot No.: 20080311.

(3) apparatus and instrument

(1) Surgical forceps, scissors, hemostats, sutures.

(2) Electric heating portable pressure steam sterilizer: the model is as follows: YXQ SG41 and 280 from Shanghai Nuclear Instrument plant.

(3) Electric heating constant temperature incubator: medical instrument factories of yellow stone city in Hubei province.

(4) Precision analytical balance: model 13P110S

The method comprises the following steps:

(1) dose setting: the dosage for human use is as follows: 32g crude drug/60 Kg body weight/day, i.e. 0.037g ointment/Kg body weight/day. The large dose is 0.444g extract/Kg (about 12 times of the clinical dosage), the medium dose is 0.222g extract/Kg (about 6 times of the clinical dosage), and the small dose is 0.111g extract/Kg (about 3 times of the clinical dosage).

(2) Route of administration and volume:

all the drugs were orally administered by gavage, 10ml/kg body weight, and the model control group was given the same volume of distilled water.

(3) Test method

On the day of the experiment, animals were anesthetized with ether, a small opening was cut in the chest, and 2 sterilized cotton balls (20 mg. + -. 1mg per cotton ball, autoclaved) were implanted under the skin of the rat's bilateral axillae, respectively, to perform molding. After 2 hours, anesthetizing animals are in a clear state, dividing rats into 5 groups according to weight, namely, a model group, an aspirin group of 250mg/kg, a traditional Chinese medicine composition high, medium and low dose group, and all the groups except a model control group of 11 animals are 12 animals, performing intragastric administration according to the weight, feeding distilled water to the model group once a day for 10 days continuously, performing cervical dislocation and sacrifice on the 11 th day, taking out cotton balls, placing the cotton balls in an incubator at 60 ℃ for 12 hours, taking out the cotton balls, weighing, and subtracting the weight of the original cotton balls to obtain the net weight of the granulation.

Data statistics

The net granulation weight value and the granulation coefficient of each animal are expressed by mean coefficient +/-standard deviation, the tested drug group and the positive control group are compared with the model control group, and the result is statistically analyzed by the spss.

The results are shown in Table 4.

TABLE 4 influence of the Chinese medicinal composition on the anti-inflammatory and repercussive action of rats (Cotton ball granuloma test)

Note: compared to the model group, 1) P < 0.05; 2) p is less than 0.01; 3) p is less than 0.001

The results show that: after the operation is carried out for 10 days, the positive medicine group, the traditional Chinese medicine composition with large dosage and the medium dosage group can reduce the weight of the cotton ball granulation compared with the model group, and has the function of reducing the cotton ball granulation coefficient.

The pathogenesis of pharyngitis caused by upper respiratory tract infection is related to chronic nonspecific inflammation, infection, environment, physicochemical factors and the like, and an implanted cotton ball granuloma rat model is a common model for researching nonspecific inflammation. Research results show that the product can reduce the weight of cotton ball granulation, has a function of reducing the cotton ball granulation coefficient, and prompts that the product has a certain anti-inflammatory effect.

1.1.3 Experimental observation of the product on irritable cough of mice caused by sulfur dioxide

Test materials:

(1) animal(s) production

KM mice, male, 18-22g in weight, supplied by Beijing Wittiulihua laboratory animal technology Co., Ltd., certification number: SCXK (Jing) 2007 + 0001; the animal is bred in the animal center (shielded environment) of the institute of traditional Chinese medicine of the Chinese academy of traditional Chinese medicine, and the license number is as follows: SYXK (Kd) 2005-0028; qing II mouse material, license number: SCXK (Jing) 2005-.

(2) Drugs and chemical agents

The tested drugs are: the Chinese medicinal composition extract is brown yellow powder, is provided by the preparation room, and has the following batch number: 090305.

positive control drug: ramus syrup (brown solution), specification: 100 ml/bottle, manufactured by tai chi group Chongqing Fuling pharmaceutical Co., Ltd, batch number: 08121535.

chemical reagents: sodium sulfite was analytically pure, purchased from beijing chemicals, lot number: 940723, respectively; sulfuric acid (analytically pure) was purchased from Beijing Chemicals II. Batch number: 901120.

(3) instrument for measuring the position of a moving object

250ml side-mouth flask, like a leather hose, a bladder, a separating funnel, a piston, etc.

The method comprises the following steps:

(1) dose setting

The tested drugs are: the dosage for human is 32g crude drug/60 Kg body weight/day, i.e. 0.5g crude drug/Kg body weight/day. The large dose is 0.74g extract/Kg (about 20 times of the clinical dosage), the medium dose is 0.37g extract/Kg (about 10 times of the clinical dosage), and the small dose is 0.185g extract/Kg (about 5 times of the clinical dosage).

Positive drugs: acute bronchitis syrup, 7.5ml/kg, (about 10 times the amount used in clinical patients).

(2) Route of administration and volume

All the drugs were orally administered by gavage, 20ml/kg body weight, and the model group was given the same volume of distilled water.

(3) Preparation of sulfur dioxide gas

A250 ml flask with a side opening is used, the side opening is connected with a bladder by a rubber tube, and sodium sulfite is placed in the flask. A separating funnel is plugged in the flask stopper, and concentrated sulfuric acid is put in the separating funnel. And opening a piston of the separating funnel, and dropping concentrated sulfuric acid to generate sulfur dioxide gas in the bottle, and storing the gas in the bladder for later use.

(5) Test method

The mice are randomly divided into 5 groups, namely a model group, a traditional Chinese medicine composition group of 0.185, 0.37, 0.74g/kg and an acute bronchitis syrup group of 7.5 ml/kg; the liquid medicine is respectively infused according to the weight of 20ml/kg, and the control group is infused with distilled water. The preparation is administered once daily for 5 days. 1 hour after the last administration, the mice were placed in a sealed bell jar (5000ml) and 15ml SO2 gas was injected. Immediately, the time was counted, stimulated for 2 minutes, and the number of coughs in mice was recorded for each period of 0-5, 6-10, 11-15, 16-20 minutes and the total number of coughs was calculated.

Data statistics

The mean, SD, and one-way analysis of variance with the model groups using SPSS software were calculated for each group and the results are shown in Table 5.

Table 5: chinese medicinal composition for treating SO₂Effect of cough-inducing mice (X + -SD)

Note: 1) represents P <0.05 compared to the model group; 2) p <0.01

The results show that: the high and low dosage groups of the product can obviously prolong the cough incubation period of model animals, and can reduce the cough frequency in the time period of 0 second to 5 seconds, and the high and medium dosage groups of the product can obviously reduce the total cough frequency of the model animals.

The sulfur dioxide method is a common method for observing the cough relieving effect of the medicine, and patients with pharyngitis are basically accompanied by obvious symptoms of throat itching and dry cough in clinic.

1.1.4 the product is used for in vivo antivirus test of mice

Test materials:

(1) animal(s) production

ICR mice, male, 12 + -2 g, purchased from Peking Wittisley laboratory animals technologies, Inc. License number: SCXK (Jing) 2006-; the animal is bred in the animal center (in a barrier environment) tested by the institute of traditional Chinese medicine of the Chinese academy of traditional Chinese medicine, and the license number is as follows: SYXK (Kd) 2005-0028; qing II mouse material, license number: SCXK (Jing) 2005-.

(2) Drugs and agents

The tested drugs are:

the product of example 4.

Shuanghuanglian oral liquid, Sanjing pharmaceutical company production lot number: 09092541.

virus: influenza virus subtype A, mouse lung adapted strain, purchased from viral institute of preventive medicine academy of sciences of China, and stored in refrigerator at-70 deg.C in our institute for use.

Ether Beijing chemical plant batch number: 090304.

(3) instruments and apparatus:

1ml of syringe, surgical forceps, scissors and weighing paper.

Type of precision analytical balance: 13P110S

The method comprises the following steps:

(1) dose setting:

Positive drugs: the dosage of the Shuanghuanglian oral liquid for adults is 1ml/kg/d, and the dosage of the Shuanghuanglian oral liquid for human clinical use is 10 times that of the Shuanghuanglian oral liquid for human clinical use, and is 10 ml/kg/d.

(2) Route of administration and volume:

both the oral gavage administration and the oral gavage administration are carried out, and 20mL/kg body weight, and the same volume of distilled water is given to a blank control group and a model control group.

(3) The test method comprises the following steps:

grouping administration: the 72 ICR mice were randomly divided into a blank control group, a model control group, a Shuanghuanglian oral liquid group, and a high, medium and low dosage group of the traditional Chinese medicine composition, and each group was 12 animals. The administration was performed by gavage for 5 days 1 time per day, and the amount of distilled water was the same for the normal control group and the infected group.

Molding: on day 2 of administration, mice in each group were lightly anesthetized with ether except for the normal control group, and then nasally infected with 15 LD50 influenza viruses, each 0.05 ml. The normal control group is 0.05ml of distilled water for each nose.

After the model is made, the drug is continuously administered, weighing is carried out on the sixth day, then the drug is killed, and the whole lung is dissected and taken out for weighing. And calculating the lung index and the inhibition rate.

Lung index ═ lung weight (g)/body weight (g) ] x 100

And (3) data statistics:

the lung indexes of each group of animals are expressed by mean coefficients plus or minus standard deviations, the tested drug group and the positive control group are compared with the model control group, and the results are subjected to statistical analysis by a single-way variance of a sps software. The results are shown in Table 6.

TABLE 6 inhibition of influenza virus pneumonia in mice (X + -SD)

Note: 1) p is less than 0.05; 2) p is less than 0.01

The results show that: the lung index of model animals can be obviously reduced by each dose group of the product.

The test shows that the product can obviously reduce the virus invasion to the respiratory system of the model animal, and has the function of directly inhibiting the virus to a certain extent, which indicates that the product has a certain antiviral function.

1.2 toxicology Studies

The product is six kinds of Chinese medicine, and is prepared through acute toxicity test and long-term toxicity test according to medicine registering regulation, with clinical application period of 5-15 days, so that the animal long-term toxicity test is selected as 90 days.

1.2.1 acute toxicity test

The test is used for carrying out acute toxicity test research of the product, and observing the acute toxicity reaction condition of a mouse in an observation period of 14 days after the mouse is administered with the drugs with the maximum concentration and the maximum volume twice in one day, and mainly observing the appearance, behavior, respiratory state, body position and posture, reaction to stimulation, toxic reaction, death condition and the like; the body weight and the food intake are measured in the observation period, and compared between the control group and the tested drug group with the same sex, so that reference is provided for drug development and clinical test.

Within 14 days of observation, the animals survived, and no obvious abnormality was found in the appearance, behavior, respiratory state, posture and posture, response to stimulation, and gross autopsy of the mice. After the mice are given with 24.0g of the dry paste powder/kgBW solution, the food intake on the day of administration is obviously lower than that of the same sex of a control group, but the food intake is not different from that of the same sex of the control group from day 4 to day 14; the body weights of mice in day 4, day 7 and day 14 administration groups were not significantly different from those of mice in the same sex control group.

The maximum dosage of the extract powder/kgBW in 1 day of the mouse is 24g, which is equivalent to 648 times of the clinical human dosage (0.037g of extract/kgBW). No obvious toxic reaction is seen.

1.2.2 Long-term toxicity test in rats

The purpose of the experiment is to observe the possible toxic reaction and the nature and degree of the toxic reaction, the development and the recovery condition of the toxic reaction when the SD rat is repeatedly administrated after 90 days; determining a toxic dose effect relationship; determining the reversibility of the target organ of toxic effect and its damage; the nontoxic reaction dose is determined, and reference data is provided for dose design of clinical safe medication and clinical toxic and side effect monitoring.

The method comprises the following steps: SD rats 80 in half male and female. Divided into three dosage groups of low, medium and high dosage, the dosage is respectively 6.25, 12.5, 25 g.kg^-1(crude drug) corresponding to the clinical human-intended dose (0.53g crude drug/kg)^-1) 12, 24 and 48 times of the total amount of the water, and a control group is additionally arranged and drinking water is given. The volume of each group is 1.0ml 100g^-1The daily dose is divided into 1 time for input. The administration is 6 days per week, no administration is given on weekdays, and the administration is continued for 90 days. The recovery period was 4 weeks. The detection indexes comprise animal general conditions, body weight, food intake, blood biochemistry and pathology related examination and the like.

As a result: no obvious abnormality is observed beside the cage during the administration period. The food intake of animals in each group increases with the weight gain, and the administration group has no obvious difference compared with the control group. There was no significant change in the hematological index and plasma prothrombin time between twelve weeks of administration and four weeks of withdrawal compared to the control. Biochemical index thirteen biochemical indexes and three electrolytes in twelve-week groups of administration have no obvious change compared with a control group. Thirteen biochemical indexes and three electrolytes of each group of animals have no obvious difference after stopping the medicine for four weeks. Twelve weeks after administration and four weeks after withdrawal, the urine eight detection results and organ coefficients of each group have no obvious change compared with the control group. Histopathological examination results suggest: at the end of administration and recovery period, compared with control group, the tested organs of the Chinese medicinal composition have no toxic pathological changes related to the tested object.

And (4) conclusion: under the test condition, the SD rat intragastric administration traditional Chinese medicine composition is comprehensively judged to have toxicity tests repeatedly administered for 90 days by 6.25g of crude drug/kg, 12.5g of crude drug/kg and 25g of crude drug/kg (equal to 12, 24 and 48 times of clinical dosage) to animals under the common conditions of weight increase, food intake, hematology detection, blood biochemical indexes, urine detection and histopathology detection, and has no obvious abnormality compared with a control group, so that the SD rat intragastric administration traditional Chinese medicine composition has no obvious toxic reaction and has higher safety in clinical application.

Clinical data (taking the product prepared in example 4)

According to the formula and the form of decoction, 48 chronic pharyngitis acute attack stages or acute pharyngitis subjects with the age of 18-65 years and without serious complications of gravity, brain, liver, kidney and the like are observed in the department of clinic of traditional Chinese medicine of Beijing Hojingtang, the self contrast design is adopted, the subjects stop taking other throat-clearing medicines and health care products in the process of trial eating, the formula is continuously taken for 5-15 days, the symptoms are observed according to pharyngalgia, pharynx itch, dry throat, dry cough and the like, the integral is calculated according to the severity of the symptoms, and the symptom improvement rate is counted; the pharynx is examined for signs such as pharyngeal mucosa congestion, mucosa edema, and pharyngeal posterior wall lymph follicular hyperplasia, and the improvement rate is calculated according to the light, middle and heavy grades of examination results.

1. Basic data: selecting 48 cases of acute and chronic pharyngitis subjects with age of 18-65 years and without serious complications of gravity center, brain, liver, kidney, etc., and adopting self-control design. During the course of eating trial, the subject stops taking other throat-clearing medicines and health products.

2. Inclusion criteria were: the medical history comprises repeated attack history of symptoms such as pharyngalgia, pharynx itch, dry pharynx, dry cough, foreign body sensation, and speaking aggravation, and is diagnosed as acute attack stage of chronic pharyngitis or acute pharyngitis. And (4) checking: edema of pharyngeal mucosa, congestion of mucosa, hyperplasia of lymph follicles in posterior pharyngeal wall, etc.

3. Exclusion criteria: the inspection or the prior medical record data proves that the diseases are caused by certain occult lesions of nasopharynx, oropharynx, larynx, nose, larynx, esophagus, neck and the whole body; those under 18 years old or over 65 years old, pregnant or lactating women and those allergic to the product; patients with serious primary diseases such as cardiovascular, cerebrovascular, liver, kidney and hemopoietic system and mental disease are combined.

4. The evaluation standard of the curative effect is as follows: (1) and (3) observation of symptoms: detailed inquiry of medical history, major pharyngeal symptoms: the integral is calculated according to the weight of symptoms, the change of the integral and the improvement rate of the symptoms are counted, and the treatment effect judgment standard of acute pharyngitis or acute attack of chronic pharyngitis is healed: the syndrome and the check integral value are reduced to 0. The effect is shown: the main symptom and the inspection integral value are reduced by more than or equal to 2/3. The improvement is as follows: the main symptoms and the inspection integral value are reduced by more than or equal to 1/3 to < 2/3. And (4) invalidation: the reduction of the chief complaint and the examination integral value is less than 1/3. (2) Pharyngeal examination: congestion of pharyngeal mucosa, edema of mucosa, and hyperplasia of lymph follicles in posterior pharyngeal wall. And classifying the light, medium and heavy results into I, II and III grades according to the examination results, respectively recording the change of the physical signs before and after the administration, and calculating the integral and improvement rate of the physical signs. (3) And (4) safety observation: blood pressure, body temperature, pulse. (4) Efficacy determination criteria: the effect is shown: the symptoms disappear or are obviously relieved, and the pharyngeal physical signs are relieved at level II;

the method has the following advantages: the symptoms are relieved, and the pharyngeal physical signs are relieved at the level I; and (4) invalidation: no obvious change in symptoms and signs.

The treatment method comprises the following steps:

the decoction is provided by the clinic part, two bags are provided every day, 150 ml/bag, 5 days are a treatment course, and at most, three treatment courses are not exceeded.

Therapeutic results

A total of 48 subjects were observed, 17 males, 31 females, age 18 years minimum, 62 years maximum, average 45.5 years. The shortest disease course is 3 days, the longest disease course is 15 days, and the average disease course is 9 days. The administration time was 23 cases within 5 days and 25 cases within 15 days. The results are shown in tables 7 to 11. 48 subjects, 17 of the males, 31 of the females, aged 18 years minimum, 62 years maximum, on average 45.5 years. The shortest disease course is 9 days, the longest disease course is 15 days, and the average disease course is 9 days. The administration time was 23 cases within 5 days and 25 cases within 15 days. The observation of efficacy shows that the traditional Chinese medicine composition has 25 (51.50%), 14 (29.10%), 9 (18.75%) and 80.60% of total effective rate. The improvement rates of symptoms such as pharyngalgia, pharynx itch, dry pharynx, cough and the like are all about 80%, the improvement rates of mucosal congestion, edema and pharyngeal wall lymph follicular hyperplasia are all about 70%, and no obvious adverse reaction is found after the medicine is taken.

TABLE 7 general data comparison of pre-test groups

Control between groups: p > 0.05

TABLE 8 improvement of the chief complaints

TABLE 9 pharyngeal physical signs improvement

TABLE 10 other systemic symptoms

Table 11 observation of efficacy (average effective rate)

Adverse reactions: no adverse reaction is found in 48 patients after taking the medicine.

Comprehensive analysis and evaluation

(1) Quality controllability: the pharmaceutical research of the product shows that the preparation process is stable and the quality is controllable.

(2) The safety of the product is proved by toxicological test research that the product has no obvious toxic reaction and has a larger safety range in clinical application.

(3) The effective product has obvious functions of relieving cough, resisting inflammation, relieving throat back wall local damage, protecting throat mucosa from congestion, swelling, ulcer and necrosis, and resisting virus, and can eliminate pharyngitis local inflammation, relieve local pathological damage, reduce pathogenic factor invasion, and enhance the disease resistance of the body, thereby playing a certain role in treatment.

In conclusion, the product is safe and effective, and has stable process and controllable quality.

The above-described embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solutions of the present invention can be made by those skilled in the art without departing from the spirit of the present invention, and the technical solutions of the present invention are within the scope of the present invention defined by the claims.

Claims

1. The traditional Chinese medicine composition is characterized by comprising the following raw materials in parts by weight: 1-5 parts of coptis chinensis, 1-5 parts of cicada slough, 2-7 parts of wild buckwheat rhizome, 2-7 parts of folium isatidis, 6-14 parts of clam shell, 6-14 parts of talc and 1-4 parts of liquorice.

2. The traditional Chinese medicine composition according to claim 1, wherein the raw materials comprise the following components in parts by weight: 2-4 parts of coptis chinensis, 2-4 parts of cicada slough, 3-5 parts of wild buckwheat rhizome, 3-5 parts of folium isatidis, 7-9 parts of clam shell, 7-9 parts of talc and 2-3 parts of liquorice.

3. The traditional Chinese medicine composition according to claim 2, wherein the traditional Chinese medicine composition is prepared into pills, granules, tablets, capsules or oral liquid.

4. The Chinese medicinal composition according to claim 3, wherein the Chinese medicinal composition is prepared into tablets.

5. The method for preparing the traditional Chinese medicine composition of any one of claims 1-2, which is characterized by comprising the following steps: extracting Coptidis rhizoma and folium Isatidis with ethanol for 2-3 times, each time with 5-7 times of ethanol for 1-1.5 hr, mixing extractive solutions, recovering ethanol, and concentrating under reduced pressure to relative density of 1.15-1.25 to obtain ethanol extract; extracting periostracum Cicadae, rhizoma Fagopyri Dibotryis, Concha Meretricis Seu Cyclinae, Glycyrrhrizae radix and pulvis Talci with water for 2-3 times, adding 7-9 times of water for each time, extracting for 1-1.5 hr for each time, concentrating the supernatant under reduced pressure to relative density of 1.15-1.20 to obtain water extract, mixing the ethanol extract and water extract, and vacuum drying.

6. The preparation method of the traditional Chinese medicine composition of claim 4, which is characterized by comprising the following steps: extracting Coptidis rhizoma and folium Isatidis with ethanol for 2-3 times, each time with 5-7 times of ethanol for 1-1.5 hr, mixing extractive solutions, recovering ethanol, and concentrating under reduced pressure to relative density of 1.15-1.25 to obtain ethanol extract; adding water into periostracum Cicadae, rhizoma Fagopyri Dibotryis, Concha Meretricis Seu Cyclinae, Glycyrrhrizae radix and pulvis Talci, extracting for 2-3 times, adding 7-9 times of water for each time, extracting for 1-1.5 hr, concentrating the supernatant under reduced pressure to relative density of 1.15-1.20 to obtain water extract, mixing the ethanol extract and water extract, vacuum drying to obtain dry extract, pulverizing the dry extract, adding adjuvants, granulating, drying, grading, tabletting, film coating, and packaging.

7. Use of the Chinese medicinal composition of any one of claims 1-4 in the preparation of a medicament for treating pharyngitis.

8. The use of claim 7, wherein the Chinese medicinal composition is used for preparing a medicament for treating acute pharyngitis or chronic pharyngitis caused by accumulated heat in lung and stomach.