Traditional Chinese medicine for treating upper respiratory infection and preparation method thereof

Abstract

The invention discloses a traditional Chinese medicine for treating upper respiratory infection. The traditional Chinese medicine is prepared from five traditional Chinese medicinal materials including radix scutellariae, fructus forsythiae, radix isatidis, cortex phellodendri and semen oroxyli. The traditional Chinese medicine takes'exterior and interior relieving and lung and stomach simultaneous treatment 'as a formula thought, has the effects of dispelling wind, dissipating heat, clearing away heat and toxic materials, relieving sore-throat, diminishing swelling and the like, and is suitable for treating diseases caused by exogenous wind-heat or lung and stomach heat excess. A preclinical pharmacodynamic evaluation result shows that the traditional Chinese medicine disclosed by the invention has the effects of improving acute pharyngitis, resisting inflammation, easing pain, clearing heat and inhibiting bacteria, has a remarkable curative effect on acute upper respiratory infection and sphagitis, and can quickly relieve symptoms.

Description

Technical Field

[0001] The invention belongs to the field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine for upper respiratory tract infection and a preparation method thereof. Background Art

[0002] Qinqiaoyanshu Granules is a new type 1 Chinese medicine under development by Jianmin Pharmaceutical Group Co., Ltd., and is currently undergoing multicenter Phase II clinical studies. The prescription of this product is derived from the clinical experience of Nanjing Hospital of Traditional Chinese Medicine, and Jiangsu Yangtze River Pharmaceuticals later completed systematic preclinical pharmaceutical research and obtained implicit clinical trial approval from the State Food and Drug Administration.

[0003] Acute upper respiratory tract infection is a general term for acute inflammation of the nasal cavity, pharynx or larynx, including the common cold, viral pharyngitis, laryngitis, herpetic pharyngitis, pharyngoconjunctival fever, bacterial pharyngitis-tonsillitis, etc. Although most of these diseases are self-limiting, they have a high incidence rate. Among them, 70% to 80% are caused by viruses, including rhinovirus, coronavirus, adenovirus, influenza and parainfluenza virus, respiratory syncytial virus, echovirus, coxsackievirus, etc. Another 20% to 30% of upper respiratory infections are caused by bacteria. The elderly, the young, the weak, the immunocompromised or patients with chronic respiratory diseases are susceptible.

[0004] Acute upper respiratory tract infection belongs to the category of "exogenous febrile disease" in traditional Chinese medicine, which is equivalent to "cold", "acute throat paralysis", "acute throat aphonia", "acute milk moth" and other diseases in traditional Chinese medicine. Through long-term clinical research, Professor Zhou Zhongying of Nanjing University of Chinese Medicine found that except for a small number of cases of wind-cold attack, the key pathogenesis of most acute throat paralysis is "wind-heat attack, lung and stomach disease". Regarding the relationship between throat paralysis and lung and stomach, "Zhang's Medical Manual" once said that "the two orifices of the throat and pharynx are from the same abdomen, but they are transformed by different pathways. The throat is in front and is responsible for outflow, while the pharynx is in the back and is responsible for swallowing. The throat is hard and empty, connecting to the lungs, and is the path for air flow, which is responsible for outflow but not intake. The pharynx is soft and empty, connecting to the stomach, and is the path for food and drink, which is responsible for intake but not outflow." Among them, (1) the throat is a lung system. The hand Taiyin lung meridian enters the lungs and runs through the throat. The throat is the passage for lung qi. The lung is a metal organ, which is responsible for qi and breathing. The Book of Experience on Sores and Ulcers, Volume 1, says: “The throat responds to the weather and is the connection of the lungs”; (2) The pharynx is the connection of the stomach. The Book of Experience on Sores and Ulcers, Volume 1, says: “The throat responds to the weather and is the connection of the lungs”; (3) The pharynx is the connection of the stomach. The Book of Puji Fang, Volume 60, Chapter on Throat says: “The throat is born from the qi of the lungs and stomach. It is responsible for the ventilation of food and water and the path of stomach qi, so it is the connection of the stomach”.

[0005] According to the current European guidelines for the management and diagnosis and treatment of acute pharyngitis, bacterial infections of the pharynx can be complicated by acute rheumatic fever or acute glomerulonephritis. In order to prevent these complications, or if the patient wants to relieve pain as soon as possible, doctors often use antibiotics to treat them. Although in the treatment of acute pharyngitis, it is not recommended to use antibiotics in cases of mild pharyngitis, and it is recommended to use ibuprofen or paracetamol to relieve the symptoms of acute pharyngitis, the use of antibiotics by patients with acute pharyngitis is still very common. Traditional Chinese medicine has a long history of treating this disease, with diverse methods and rich experience. Many Chinese medicines for clearing heat and detoxifying have certain antibacterial effects and the effect of inactivating endotoxins. They can be called "broad-spectrum antibiotics" for the treatment of acute pharyngitis. They are effective, safe and reliable, not easy to resist, and do not disrupt the balance of the human body's internal environment and flora while killing bacteria. They also have the characteristics of diverse dosage forms and easy to carry and take. Summary of the invention

[0006] The purpose of the present invention is to provide a traditional Chinese medicine for upper respiratory tract infection and a preparation method thereof.

[0007] The traditional Chinese medicine provided by the invention is composed of five medicinal materials such as scutellaria baicalensis, forsythia suspensa, isatis root and the like, and the specific formula is as follows: 5-20 parts by weight of scutellaria baicalensis; 5-20 parts by weight of forsythia suspensa; 10-25 parts by weight of isatis root; 3-10 parts by weight of phellodendron chinense; and 2-10 parts by weight of oroxylum butterfly.

[0008] Preferably, the formula of the traditional Chinese medicine is as follows: 8-15 parts by weight of Scutellaria baicalensis; 8-15 parts by weight of Forsythia suspensa; 12-20 parts by weight of Isatis indigotica; 5-8 parts by weight of Phellodendron chinense; and 5-8 parts by weight of Oroxylum indica.

[0009] Optimally, the formula of the traditional Chinese medicine is as follows: 10 parts by weight of Scutellaria baicalensis; 12 parts by weight of Forsythia suspensa; 15 parts by weight of Isatis indigotica; 6 parts by weight of Phellodendron chinense; and 6 parts by weight of Oroxylum indica.

[0010] In the process of clinical treatment of acute upper respiratory tract infection, Professor Zhou Zhongying, a master of traditional Chinese medicine, is good at using "resolving both the exterior and interior, and treating the lungs and stomach at the same time" as the main formula compatibility idea. In the formula, Forsythia suspensa is pungent, bitter, slightly cold, clears heat and detoxifies, reduces swelling and disperses knots, and is good at dispersing wind-heat and relieving the exterior; Scutellaria baicalensis is bitter, cold, clears heat and dries dampness, purges fire and detoxifies, and is good at clearing and purging heat in the interior. The two medicines are used together to play the role of relieving the exterior and clearing the interior, and are the main medicine. Isatis root clears heat and detoxifies, relieves sore throat and reduces swelling, and is good at acute tonsillitis. The addition of Phellodendron chinense clears heat and dries dampness, purges fire and removes steaming, which not only strengthens the function of clearing heat and detoxification, but also is good at clearing latent heat. These two medicines are the minister medicines. Oroxylum indica is sweet, cool, and enters the lung, liver, and uterus meridians. It not only improves the function of clearing the lung and relieving sore throat, but also takes into account cough and phlegm, soothing the liver and stomach, and is an adjuvant medicine. The combination of these medicines can relieve both the exterior and interior, and treat both the lungs and stomach. It mainly focuses on dispersing wind and heat, clearing away heat and detoxifying, relieving sore throat and reducing swelling. It also has the functions of relieving cough and resolving phlegm, soothing the liver and harmonizing the stomach. It is suitable for patients with acute throat paralysis caused by wind-heat, and lung and stomach diseases.

[0011] Furthermore, the pharmacodynamics of traditional Chinese medicine was evaluated for acute pharyngitis, anti-inflammatory, analgesic, antipyretic and antibacterial effects. The results showed that Qinqiao Yanshu Granules have the effects of improving acute pharyngitis, anti-inflammatory, analgesic, antipyretic and antibacterial, providing a basis for its clinical use in upper respiratory tract infections and pharyngitis. The specific contents are as follows: 1. Alleviating effect on acute pharyngitis model in rats An acute pharyngitis model was induced in rats by 15% ammonia spray, and the effects of Qinqiao Yanshu Granules on the pathology of rat pharyngeal tissue, blood routine and the levels of inflammatory factors in serum were investigated. The results showed that Qinqiao Yanshu Granules can significantly improve the lesions such as hyperplasia, hypertrophy, submucosal edema, and inflammation of the rat throat tissue mucosa; the NEUT content was increased and the LYM content was decreased in the blood routine examination; the content of inflammatory factor IL-6 was reduced, indicating that Qinqiao Yanshu Granules has a relieving effect on acute pharyngitis.

[0012] 2. Anti-inflammatory Effects The permeability of the peritoneal capillaries of mice increased by intraperitoneal injection of 0.8% acetic acid, and Qinqiao Yanshu granules can significantly inhibit the increase in permeability of the peritoneal capillaries of mice caused by acetic acid; the ear swelling model of mice was induced by applying 2% croton oil, and Qinqiao Yanshu granules can significantly reduce the ear swelling value of mice caused by croton oil. The above results show that Qinqiao Yanshu granules have anti-inflammatory effects.

[0013] 3. Analgesic effect A pain model was induced in mice by intraperitoneal injection of 0.8% acetic acid. The number of body twistings in the Qinqiao Yanshu granules group was significantly lower than that in the model control group, indicating that Qinqiao Yanshu granules have an inhibitory effect on the pain response of mice induced by acetic acid.

[0014] 4. Antipyretic effect The body temperature of rats increased to varying degrees after subcutaneous injection of yeast solution. After oral administration of Qinqiao Yanshu granules, the elevated body temperature of rats was reduced to varying degrees compared with the model control group, indicating that Qinqiao Yanshu granules have an antipyretic effect.

[0015] 5. In vitro antibacterial effect In vitro antibacterial experiments showed that Qinqiaoyanshu granules exhibited antibacterial and bactericidal effects on Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus epidermidis.

[0016] The present invention further provides a method for preparing the traditional Chinese medicine, which comprises three steps of water decoction extraction, alcohol precipitation and preparation molding, wherein the purpose of alcohol precipitation is to reduce the content of "impurities" in traditional Chinese medicine such as pectin, starch, tannin, protein, polysaccharide, etc., so as to enrich the active ingredients to improve the efficacy, and the specific steps are as follows: a. Boil Scutellaria baicalensis, Forsythia suspensa, Isatis indigotica, Phellodendron chinense and Oroxylum indica in water; b. Combine the decoction and filter, then concentrate to a relative density of 1.1-1.2, add ethanol after the concentrate is cooled to make the alcohol content reach 30-60%, let stand for 20-50 hours, separate the supernatant and concentrate into a clear paste; c. Add auxiliary materials to the clear paste and make it into a medicine suitable for clinical use.

[0017] Preferably, the decoction is performed 1-3 times, the amount of water added for each decoction is 5-15 times the total weight of the raw materials, and the decoction time for each decoction is 0.5-3 hours.

[0018] Further preferably, the decoction is performed twice, the amount of water added in the first decoction is 12 times the total weight of the raw materials, and the decoction time is 2 hours, and the amount of water added in the second decoction is 10 times the total weight of the raw materials, and the decoction time is 1 hour.

[0019] Preferably, in step b, the relative density is 1.12, the alcohol content is 40%, and the standing time is 24 hours.

[0020] For those skilled in the art, it is a conventional method to make a Chinese medicine paste into a medicament suitable for clinical use. The medicament described in the present invention includes any dosage form suitable for clinical use, such as tablets, granules, capsules, pills, pastes, mixtures, syrups, etc. The preferred dosage form of the present invention is granules, tablets, capsules or oral liquids, which have the advantages of low production cost and good compliance. The auxiliary materials used to prepare these dosage forms can be a combination of one or more of these pharmaceutical auxiliary materials such as sucrose, starch, dextrin, sorbitol, mannitol, lactose, carboxymethyl cellulose, water, flavoring agents, preservatives, etc.

[0021] The alcohol content involved in the present invention refers to the volume content. For example, an alcohol content of 40% means that 40 ml of ethanol is contained in every 100 ml of the liquid medicine.

[0022] The beneficial effects of the present invention are: The Chinese medicine of the present invention has significant curative effect on acute upper respiratory tract infection and pharyngitis, can quickly relieve symptoms, and relieve the pain of patients, and has no toxic side effects and high safety. The Chinese medicine raw materials are easy to obtain, the preparation process is simple, the production cost is low, it is conducive to reducing the burden on doctors and patients, and it also has the advantages of convenient administration, high compliance, etc. DETAILED DESCRIPTION

[0023] The present invention is further described in detail below in conjunction with specific embodiments.

[0024] Example 1 Prescription: Scutellaria baicalensis 277.78g; Forsythia suspensa 333.33g; Isatis indigotica 416.67g; Phellodendron chinense 166.67g; Oroxylum indica 166.67g Preparation method: The above 5 ingredients (Scutellaria baicalensis is added in boiling water) are decocted twice. The first time, 12 times the amount of water is added to the entire prescription and decocted for 2 hours. The second time, 10 times the amount of water is added to the entire prescription and decocted for 1 hour. The decoctions are combined, filtered, and the filtrate is concentrated into a clear paste. Finally, sucralose and maltodextrin are added to the clear paste in appropriate amounts, and granules are made and dried.

[0025] Properties: This product is yellow-brown to brown granules; bitter and slightly sweet.

[0026] Functions and indications: dispel wind and heat, clear away heat and detoxify, relieve sore throat and reduce swelling. Suitable for dry throat, sore throat, fever, yellow sputum caused by exogenous wind-heat, excessive heat in the lungs and stomach; acute and chronic pharyngitis, acute laryngitis, acute tonsillitis, upper respiratory tract infection with the above symptoms.

[0027] Usage and dosage: Take with boiled water. 1 bag at a time, 3 times a day. Specification: Each 1g is equivalent to 1.36g of medicinal material.

[0028] Example 2 Prescription: Same as Example 1.

[0029] Preparation method: The above 5 ingredients (Scutellaria baicalensis in boiling water) are boiled three times with water. The first time, 10 times the amount of water is added and boiled for 1 hour. The second time, 8 times the amount of water is added and boiled for 1 hour. The third time, 5 times the amount of water is added and boiled for 0.5 hours. Combine the decoctions, filter, and concentrate the filtrate into a clear paste. Finally, add appropriate amounts of sucralose and maltodextrin to the clear paste, make granules, and dry them.

[0030] Example 3 Prescription: Same as Example 1.

[0031] Preparation method: add 20 times the water to the above 5 ingredients (add scutellaria baicalensis in boiling water) and boil once for 3 hours, filter and concentrate the filtrate into a clear paste. Finally, add appropriate amount of sucralose and maltodextrin to the clear paste, make granules, and dry to obtain the product.

[0032] Example 4 Prescription: Same as Example 1.

[0033] Preparation method: The above 5 ingredients (Scutellaria baicalensis is added in boiling water) are decocted twice. The first time, 12 times the amount of water in the entire prescription is added and decocted for 2 hours. The second time, 10 times the amount of water in the entire prescription is added and decocted for 1 hour. The decoctions are combined, filtered, and the filtrate is concentrated to a relative density of 1.16 (55±5℃). After the concentrated solution is cooled, ethanol is added to make the alcohol content reach 60%. After standing for 48 hours, the supernatant is separated and concentrated into a clear paste. Finally, sucralose and maltodextrin are added to the clear paste, and granules are made and dried.

[0034] Example 5 Prescription: Same as Example 1.

[0035] Preparation method: The above 5 ingredients (Scutellaria baicalensis is added in boiling water) are decocted twice. The first time, 12 times the amount of water in the entire prescription is added and decocted for 2 hours. The second time, 10 times the amount of water in the entire prescription is added and decocted for 1 hour. The decoctions are combined, filtered, and the filtrate is concentrated to a relative density of 1.12 (55±5℃). After the concentrated solution is cooled, ethanol is added to make the alcohol content reach 40%. After standing for 24 hours, the supernatant is separated and concentrated into a clear paste. Finally, sucralose and maltodextrin are added in appropriate amounts to the clear paste, and granules are made and dried to obtain the product.

[0036] In order to evaluate the extraction effect, we took a small amount of the clear paste prepared in Examples 1-5 before adding the auxiliary materials for granulation, dried it under reduced pressure to constant weight, and then used HPLC to detect the content of the active ingredients baicalin and oroxylin B in the dry paste. The specific method of content determination is as follows: (1) Preparation of reference solution: Take an appropriate amount of baicalin reference substance, accurately weigh it, and add methanol to make a solution containing 1.0 mg of baicalin per 1 ml; take an appropriate amount of oroxylin B reference substance, accurately weigh it, and add 50% methanol to make a solution containing 0.2 mg of oroxylin B per 1 ml; accurately transfer 5 ml of baicalin reference solution and 2 ml of oroxylin B reference solution into the same 10 ml volumetric flask, add methanol to dilute to the scale, and mix well to obtain (0.5 mg of baicalin and 40 μg of oroxylin B per 1 ml).

[0037] (2) Preparation of test solution: Take an appropriate amount of dry paste, grind it into fine powder, take about 1.0 g, weigh it accurately, place it in a stoppered conical flask, accurately add 50 ml of 70% methanol, stopper it tightly, weigh it accurately, treat it with ultrasound for 20 minutes, cool it, weigh it again, make up the lost weight with 70% methanol, shake it well, filter it, and take the filtrate to obtain the solution.

[0038] (3) Accurately pipette 5 μl of the reference solution and the test solution, respectively, and inject them into the liquid chromatograph for determination. Chromatographic conditions and system suitability test: octadecylsilane bonded silica gel as filler; acetonitrile as mobile phase A, 0.2% phosphoric acid solution as mobile phase B, gradient elution; flow rate of 1.0 ml per minute; detection wavelength of 320 nm. The theoretical plate number calculated based on the baicalin peak should not be less than 10,000.

[0039] The content determination results of Examples 1-5 are shown in Table 1 below.

[0040] Table 1 Contents of baicalin and oroxylin B in dry extract

[0041] From the comparison results of Examples 1-3, it can be seen that the number, time and amount of water added for decoction have a great influence on the content of active ingredients, especially the content of orchidin B in Example 1 is significantly higher than that in Examples 2 and 3. Compared with Example 1, the contents of baicalin and orchidin B in Examples 4 and 5 are significantly increased, indicating that after alcohol precipitation treatment, the "impurities" in the dry paste are reduced and the active ingredients are effectively retained, which is beneficial to improving the efficacy of traditional Chinese medicine.

[0042] Example 6 Efficacy Verification Test This embodiment is excerpted from the registration materials of clinical approval certificate of new Chinese medicine submitted to the State Food and Drug Administration and approved.

[0043] 1. Test materials 1.1 Drugs under investigation Qinqiaoyanshu granules dry paste powder is provided by Jiangsu Longfengtang Chinese Medicine Co., Ltd. of Yangtze River Pharmaceutical Group, batch number: 21011411, and each gram of dry paste powder contains 4.13g of raw medicine. 1.2 Positive control drug Dexamethasone acetate tablets, Zhejiang Xianjun Pharmaceutical Co., Ltd., batch number: 200421. Ibuprofen granules, Harbin Pharmaceutical Group Shiyitang Pharmaceutical Factory, batch numbers: 1906607, 2003611. Aspirin effervescent tablets, AstraZeneca Pharmaceuticals Co., Ltd., batch number: 1908221.

[0044] 1.3 Dosage design The proposed clinical dosage of Qinqiaoyanshu Granules is 49 g of crude drug per day. Calculated based on the equivalent dose ratio of animal to human surface area, the clinical equivalent dose for rats is 4.41 g of crude drug / kg, which is set as the medium dose. The high, medium and low doses for rats are 8.82, 4.41 and 2.21 g of crude drug / kg, respectively; the clinical equivalent dose for mice is 6.37 g of crude drug / kg, and the experimental doses for mice are 25.48, 12.74, 6.37 and 3.19 g of crude drug / kg.

[0045] Dexamethasone acetate tablets, the clinical daily dose is calculated as 16 tablets, the specification is 0.75 mg per tablet, and the equivalent dose is converted based on the ratio of animal surface area to human surface area, and the twice of the clinical equivalent dose in rats is 2.16 mg / kg. Ibuprofen granules, the clinical daily dose is calculated as 0.8 grams, and the equivalent dose is converted based on the ratio of animal surface area to human surface area, and the clinical equivalent dose in mice is 0.10 g / kg, and the twice of the clinical equivalent dose in mice is 0.21 g / kg. Aspirin effervescent tablets, the clinical daily dose is calculated as 2.0 grams, and the equivalent dose is converted based on the ratio of animal surface area to human surface area, and the clinical equivalent dose in rats is 0.18 g / kg.

[0046] 1.4 Animals SD rats, SPF grade, weighing 180-200g; ICR mice, SPF grade, weighing 18-20g; half male and half female, provided by Beijing Weitonglihua Laboratory Animal Technology Co., Ltd., Animal Production License No.: SCXK (Beijing) 2016-0006. They were kept in a barrier environment, and the Laboratory Animal Use License No.: SYXK (Beijing) 2019-0003.

[0047] 1.5 Dosage and route of administration The dosage for rats was 10 mL / kg, and that for mice was 20 mL / kg, and the drug was administered by intragastric gavage.

[0048] 1.6 Bacteria Pseudomonas aeruginosa, strain number: 10104, batch number: 2a28-2; Staphylococcus aureus, strain number: 26003, batch number: 9a3-1; Escherichia coli, strain number: 44102, batch number: 3a33-1; purchased from China Food and Drug Administration Center for Medical Bacteria. Staphylococcus epidermidis, strain number: 12228, purchased from Zhengzhou Jingsiwei Chemical Co., Ltd.

[0049] 1.7 Reagents Ammonia water, Beijing Chemical Plant, batch number: 20180808; IL-6 ELISA Kit, WUHAN HUAMEI BIOTECH., LTD, batch number: E13037834; TNF-α ELISA Kit, WUHAN HUAMEI BIOTECH.,LTD, batch number: E14037853; Evansland, Solarbio, batch number: NO.619N041; acetic acid (glacial acetic acid), Beijing Chemical Plant, batch number: 20170401; sodium chloride injection, Shijiazhuang Siyao Co., Ltd., batch number: 2008183203; croton oil, Beijing Coupling Technology Co., Ltd., batch number: 150318; anhydrous ethanol, Sinopharm Chemical Reagent Co., Ltd., batch number: 20181009; ether, Sinopharm Chemical Reagent Co., Ltd., batch number: 20181210; high-activity dry yeast, Angel Yeast Co., Ltd., production date: 2021 / 01 / 12; nutrient agar medium, Beijing Aoboxing Biotechnology Co., Ltd., batch number: 20190418; nutrient broth medium, Beijing Aoboxing Biotechnology Co., Ltd., batch number: 20190516.

[0050] 2. Test methods and results 2.1 Effects on the rat acute pharyngitis model 2.1.1 Test methods Healthy SD rats, SPF grade, weighing 180-200g were selected and randomly divided into normal control group and modeling group according to their weight. The modeling group was sprayed with 15% ammonia water on the pharynx, once in the morning and afternoon, 3 presses each time with a laryngeal sprayer, for 3 consecutive days, and the normal control group was sprayed with the same amount of pure water. On the 4th day, the modeling group was randomly divided into model control group, high-, medium- and low-dose groups of Qinqiao Yanshu granules (8.82, 4.41, 2.21 g crude drug / kg) and positive control drug dexamethasone acetate tablets group according to their weight, with 10 rats in each group, half male and half female. Each group was gavaged and administered with equal volume of pure water (10mL / kg body weight) by gavage, once a day, for 5 consecutive days. One hour after the last administration, the rats were anesthetized, and EDTAK2 anticoagulated blood and non-anticoagulated blood were collected. The anticoagulated blood was used for white blood cell count and classification. The non-anticoagulated blood was centrifuged to separate the serum, and the ELISA kit was used to determine the levels of inflammatory factors IL-6 and TNF-α in the serum. The pharynx was taken and fixed in 10% formalin solution for pathological histological examination.

[0051] 2.1.2 Test results 2.1.2.1 Results of rat pharyngeal histopathological examination The rats in the normal control group had no hyperkeratosis in the pharyngeal mucosa, no hyperplasia or thickening of the mucosal epithelial cells, and no apoptosis of the cells; there was no edema or inflammation in the submucosal muscle tissue, no hypertrophy of the glands, and the tissue structure was normal. The rats in the model control group had hyperkeratosis in the pharyngeal mucosa, obvious hyperplasia and enlargement of the mucosal epithelial cells, and apoptotic cells; there was inflammatory exudation in the submucosal layer, mainly lymphocytes, denucleated mast cells, a small number of eosinophils and neutrophilic lobed nuclear cells; the submucosal lamina propria tissue was congested and edematous, and the glands were swollen; there was edema in the submucosal muscle layer, the muscle cells were broken, and there was a small amount of inflammation around them.

[0052] The lesions of mucosal hyperplasia, hypertrophy, submucosal edema, inflammation and other lesions in the throat tissue of rats in the high, medium and low dose groups of Qinqiao Yanshu Granule and the positive drug dexamethasone acetate tablets group were alleviated to varying degrees compared with the model control group. The results of the rank sum test showed that the high dose group of Qinqiao Yanshu Granule and the dexamethasone acetate tablets group could significantly alleviate the pharyngeal lesions, which were significantly different from the model control group. The results are shown in Table 2.

[0053] Table 2 Statistical table of pathological observation of pharyngeal tissue under microscope in rats with acute pharyngitis model

[0054] Note: Rank sum test was used to compare each group with the model control group. Appendix: Microscopic Observation Standards “-”: There was no congestion or edema in the rat pharyngeal mucosa, no thickening of the mucosal epithelium, no hypertrophy of the submucosal glands, and the tissue structure was normal. “+”: There was no hyperplasia or edema in the rat pharyngeal mucosal epithelium, no thickening of the mucosal epithelium, mild congestion under the mucosal epithelium, and no inflammation. “++”: There is mild localized hyperplasia of the rat pharyngeal mucosal epithelium, congestion under the mucosal epithelium, and a small amount of inflammatory exudate. “+++”: The rat pharyngeal mucosal epithelium is hyperkeratotic and segmental, the mucosal epithelial cells are hyperplastic and thickened, the subepithelial muscle layer is swollen, there is muscle tissue rupture, and there is a small amount of inflammation. “++++”: The rat pharyngeal mucosal epithelium is hyperkeratotic, with epithelial hyperplasia and spike formation. There is inflammatory exudate under the mucosal epithelium, glandular hypertrophy, mainly lymphocytes, a small number of lobed nuclear cells, and a wide area of ​​lesions. 2.1.2.2 Effect on WBC count and classification Compared with the model control group, the NEUT (%) (neutrophil percentage) of the high- and medium-dose Qinqiao Yanshu granules groups and the dexamethasone acetate tablets group was significantly increased, and the LYM (%) (lymphocyte percentage) was significantly decreased, which was significantly different from the model control group. The results are shown in Table 3.

[0055] Table 3 Effects of Qinqiaoyanshu Granules on the white blood cell count and classification in rats with acute pharyngitis model (X ± SD)

[0056] Note: *P<0.05, **P<0.01 compared with the model control group.

[0057] 2.1.2.3 Effects on the levels of inflammatory factors in rat serum The IL-6 content in the serum of the model control group was significantly higher than that of the normal group (P<0.01). The IL-6 content in the serum of the high-dose Qinqiaoyanshu granules (8.82 g crude drug / kg) and dexamethasone acetate tablets groups was significantly lower than that of the model control group, which was significantly different from the model control group. The results are shown in Table 4. Table 4 Effects of Qinqiaoyanshu Granules on inflammatory factors in rat acute pharyngitis model (X ± SD)

[0058] Note: Compared with the model control group, *P<0.05, **P<0.01.

[0059] 2.2 Anti-inflammatory experiments 2.2.1 Effects on acetic acid-induced increase in peritoneal capillary permeability in mice 2.2.1.1 Test methods Fifty healthy ICR mice, SPF grade, weighing 18-20g, were randomly divided into 5 groups according to body weight, including model control group, three dose groups of Qinqiaoyanshu granules (12.74, 6.37, 3.18 g crude drug / kg) and positive control ibuprofen granules group, 10 mice in each group, half male and half female. Each group was given corresponding drugs by gavage, the model control group was given the same volume of pure water (0.2mL / 10g body weight) by gavage once a day, the ibuprofen granules group started to be given drugs on the second day, and the drugs were given for 2 consecutive days, and the other groups were given drugs for 3 consecutive days. One hour after the last dose, 0.5% Evans blue solution (0.1 mL / 10 g) was injected through the tail vein. Ten minutes later, 0.8% acetic acid solution (0.1 mL / 10 g) was injected intraperitoneally. Ten minutes later, 5 mL of normal saline injection was injected intraperitoneally. The mice were killed, the abdomen was gently rubbed 50 times, the skin was cut open, the peritoneal fluid was aspirated with a pipette, centrifuged, and the supernatant was measured for absorbance at a wavelength of 590 nm. The t-test was used for comparison among the groups, and the inhibition rate was calculated.

[0060]

[0061] 2.2.1.2 Test results The tested doses of Qinqiaoyanshu granules (12.74, 6.37, 3.18 g crude drug / kg) showed an inhibitory trend on the increase in peritoneal capillary permeability induced by acetic acid in mice, and the absorbance of peritoneal fluid was lower than that of the model control group. The results are shown in Table 5. Table 5 Effect of Qinqiaoyanshu Granules on increased permeability of peritoneal capillaries (X ± SD) (administration for 3 days)

[0062] Note: Compared with the model control group, *P<0.05, **P<0.01.

[0063] 2.2.2 Effects of croton oil on ear swelling in mice 2.2.2.1 Test methods Fifty healthy ICR mice, SPF grade, weighing 18-20g, were randomly divided into 5 groups according to body weight, including model control group, three dose groups of Qinqiaoyanshu granules (25.48, 12.74, 6.37 g crude drug / kg) and positive control drug ibuprofen granules group, with 10 mice in each group, half male and half female. Each group was given the corresponding drug by gavage, the model control group was given the same volume of pure water (0.2mL / 10g body weight) by gavage once a day, the ibuprofen granules group started to be given on the 4th day, and continued to be given for 3 days, and the other groups continued to be given for 6 days. After the last administration, 70μL / mouse of 3% croton oil was applied to the front and back of the right ear, and the left ear was used as a control. The mice were killed 4 hours after inflammation, and the two ears were cut along the auricle baseline. The ear pieces were punched out at the same position with an 8mm diameter ear puncher, and the weight difference between the left and right ear pieces (i.e., ear swelling value) was calculated by electronic balance. The t test was used for inter-group comparison, and the inhibition rate was calculated.

[0064]

[0065] 2.2.2.2 Test results The doses of 25.48, 12.74, and 6.37 g crude drug / kg of Qinqiaoyanshu Granules and the positive drug ibuprofen granules can significantly reduce the ear swelling value of mice induced by croton oil, which is significantly different from the model control group, indicating that Qinqiaoyanshu Granules have anti-inflammatory effects. The results are shown in Table 6.

[0066] Table 6 Effect of Qinqiaoyanshu Granules on Croton Oil-induced Ear Swelling in Mice (X ± SD)

[0067] Note: Compared with the model control group, *P<0.05, **P<0.01.

[0068] 2.3 Analgesia experiment 2.3.1 Test methods Fifty healthy SPF ICR mice weighing 18-22g were randomly divided into model control group, three dose groups of Qinqiaoyanshu granules (12.74, 6.37, 3.18g crude drug / kg), and positive control ibuprofen granules group, with 10 mice in each group, half male and half female. Each group was gavaged, and the model control group was gavaged with an equal volume of purified water once a day. The ibuprofen granules group started to be administered on the third day and continued for 3 days. The rest of the groups were administered for 5 consecutive days. One hour after the last administration, 0.8% acetic acid 0.1ml / 10g was injected intraperitoneally, and the typical number of twisting of mice (extending hind limbs, abdominal contraction and twisting body) within 20min was observed. The t test was used for inter-group comparison, and the inhibition rate was calculated.

[0069]

[0070] 2.3.2 Test results The number of twistings of Qinqiaoyanshu Granules (12.74 g crude drug / kg) and positive drug ibuprofen granules was significantly lower than that of the model control group, and there was a significant difference compared with the model control group, indicating that Qinqiaoyanshu Granules has an inhibitory effect on the pain response of mice induced by acetic acid. The results are shown in Table 7. Table 7 Effects of Qinqiaoyanshu Granules on pain response of mice induced by acetic acid (X ± SD) (administered for 6 days)

[0071] Note: Compared with the model control group, **P<0.01, *P<0.05 2.4 Antipyretic experiment 2.4.1 Test methods Healthy SD rats, SPF grade, weighing 180-200g were selected. The rectal temperature of the animals was measured once a day for 4 days before the experiment. The animals were fasted for 10 hours before the experiment, and the rectal temperature of the animals was measured twice in the morning of the experiment as the basic rectal temperature. 20% yeast solution was injected subcutaneously on the back to induce fever (2mL / 100g). The rectal temperature was measured 5 hours later, and those with an increase of more than 0.8℃ were used for the experiment. They were randomly divided into model control group, high-, medium-, and low-dose groups of Qinqiaoyanshu granules (8.82, 4.41, and 2.21 g crude drug / kg) and positive control aspirin effervescent tablets group, with 10 rats in each group, half male and half female. Each group was gavaged with drugs, and the model control group was gavaged with an equal volume of pure water (10mL / kg body weight). The rectal temperature was measured once 1, 2, 3, and 4 hours after the drug, and the difference between the rectal temperature measured at different times and the basic rectal temperature was used as an indicator of body temperature change, and statistical analysis was performed using t-test.

[0072] 2.4.2 Test results The results showed that the body temperature of rats increased to varying degrees after subcutaneous injection of yeast solution, and the increase value above 0.8℃ was selected for the experiment. After intragastric administration, the increase in body temperature of rats in each administration group was reduced to varying degrees compared with the model control group. The high-dose group of Qinqiaoyanshu granules (8.82 g crude drug / kg) could significantly inhibit the increase in body temperature of rats caused by yeast 2-4 hours after administration, which was significantly different from the model control group. The positive control drug aspirin effervescent tablets could significantly inhibit the increase in body temperature of rats caused by yeast within 4 hours after administration, indicating that Qinqiaoyanshu granules have an antipyretic effect. The results are shown in Table 8.

[0073] Table 8 Effect of Qinqiaoyanshu Granules on yeast-induced fever in rats (X ± SD)

[0074] Note: Compared with the model control group, *P<0.05, **P<0.01.

[0075] 2.5 In vitro antibacterial experiment 2.5.1 Test method: Liquid dilution method The day before the test, each bacterium was inoculated in broth medium for 16 hours overnight. During the test, several 96-well cell culture plates were taken and 100 μL / well of broth medium was added first. 100 μL / well of drug sample was added to the first row of each plate, mixed, 100 μL was pipetted down to the second row, and mixed and moved down in the same way, and repeated until the eighth dilution, mixed, and discarded 100 μL / well. The concentration of the first row was 500 mg / mL, the second row was 250 mg / mL, and the concentrations of the third, fourth, fifth, sixth, seventh, and eighth rows were 125, 62.5, 31.2, 15.6, 7.8, and 3.9 mg / mL, respectively. The enriched bacteria were turbidized into McFarland No. 1 tube (300 million bacteria / mL), and then diluted 100 times, 10 μL of each bacterium was pipetted into each row, and four columns were added for each bacterium. Positive control and culture medium control were performed in parallel. Add 100 μL / well of broth to each plate. Incubate each plate at 37℃ for 24 hours, observe the results under a microscope, and record the minimum inhibitory concentration (MIC). Take 5 μL from each MIC well and transfer it to an agar plate. Incubate at 37℃ for 24 hours, and observe whether there is bacterial growth to determine the minimum bactericidal concentration (MBC).

[0076] 2.5.2 Test results In vitro antibacterial experiments showed that the extract of Qinqiaoyanshu granules showed antibacterial and bactericidal effects on standard strains of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus epidermidis. The results are shown in Table 9.

[0077] Table 9 Antibacterial test results of Qinqiaoyanshu granules

[0078] In summary, the pharmacodynamic experiment of Qinqiao Yanshu Granules showed that Qinqiao Yanshu Granules have the effects of improving acute pharyngitis, anti-inflammatory, analgesic, antipyretic, and antibacterial. It is intended to be used for acute upper respiratory tract infection, with acute pharyngitis, acute laryngitis, acute tonsillitis, or colds accompanied by fever, sore throat, yellow sputum, etc. as the main manifestations, as well as acute exacerbation of chronic pharyngitis. This study provides a pharmacodynamic test basis for its clinical application.

[0079] Example 7 From May to July 2020, 30 patients with acute upper respiratory tract infection were treated in Nanjing Hospital of Traditional Chinese Medicine. Among them, 17 patients were treated with 3-day Qinqiao Yanshu granules, and 13 patients were treated with 3-day Qinqiao oral liquid (National Medicine Standard Z10970135). The average age of patients in the Qinqiao Yanshu granule group was 31.6 years old, including 9 male patients (52.9%) and 8 female patients (47.1%); the average age of patients in the Qinqiao oral liquid group was 29.5 years old, including 7 male patients (53.8%) and 6 female patients (46.2%).

[0080] The symptoms and signs of the patients before and after treatment were evaluated, among which fever, sore throat, and red and swollen throat were the main symptoms, which were scored as 0, 2, 4, and 6 points as no symptoms, mild, moderate, and severe, respectively; cough, thick sputum, thirst, dry stool, yellow urine, and thick and greasy tongue coating were scored as 0, 1, 2, and 3 points as no symptoms, mild, moderate, and severe, respectively.

[0081] Table 10 Symptom scores before and after treatment

[0082] The above results show that Qinqiao Yanshufang has a significant effect on improving the symptoms and signs of the throat of acute upper respiratory tract infection, and its efficacy in improving the total score of symptoms is better than that of the similar Chinese medicine Qinqiao oral liquid that has been marketed. No adverse events were reported in the 30 subjects enrolled during the trial.

[0083] Example 8 A traditional Chinese medicine for upper respiratory tract infection, the prescription and process are as follows: Prescription: 450g of Scutellaria baicalensis, 450g of Forsythia suspensa, 300g of Isatis indigotica, 150g of Phellodendron chinense, and 150g of Oroxylum bungeana. The preparation method is the same as that of Example 5.

[0084] Example 9 A traditional Chinese medicine for upper respiratory tract infection, the prescription and process are as follows: Prescription: Scutellaria baicalensis 240g; Forsythia suspensa 160g; Isatis root 240g; Phellodendron chinense 160g; Oroxylum indica 160g The preparation method is the same as that of Example 5.

Claims

1. A Chinese medicine for upper respiratory tract infection, characterized in that The invention is prepared from the following raw materials in parts by weight: 5-20 parts of scutellaria baicalensis, 5-20 parts of forsythia suspensa, 10-25 parts of isatis root, 3-10 parts of phellodendron amurense and 2-10 parts of oroxylum bungeana.

2. The Chinese medicine according to claim 1, characterized in that The invention is prepared from the following raw materials in parts by weight: 8-15 parts of scutellaria baicalensis, 8-15 parts of forsythia suspensa, 12-20 parts of isatis root, 5-8 parts of phellodendron amurense and 5-8 parts of oroxylum bungeana.

3. The Chinese medicine according to claim 1, characterized in that The invention is prepared from the following raw materials in parts by weight: 10 parts of scutellaria baicalensis, 12 parts of forsythia suspensa, 15 parts of isatis root, 6 parts of phellodendron amurense and 6 parts of oroxylum butterfly.

4. A method for preparing the Chinese medicine according to any one of claims 1 to 3, characterized in that: The following steps are involved: a. Boil Scutellaria baicalensis, Forsythia suspensa, Isatis indigotica, Phellodendron chinense and Oroxylum indica in water; b. Combine the decoction and filter, then concentrate to a relative density of 1.1-1.2, add ethanol after the concentrate is cooled to make the alcohol content reach 30-60%, let stand for 20-50 hours, separate the supernatant and concentrate into a clear paste; c. Add auxiliary materials to the clear paste and make it into a medicine suitable for clinical use.

5. The method according to claim 4, characterized in that: The decoction is performed 1-3 times, the amount of water added for each decoction is 5-15 times the total weight of the raw materials, and the decoction time for each decoction is 0.5-3 hours.

6. The method according to claim 5, characterized in that: The decoction is performed twice, the amount of water added in the first decoction is 12 times the total weight of the raw materials, and the decoction time is 2 hours, and the amount of water added in the second decoction is 10 times the total weight of the raw materials, and the decoction time is 1 hour.

7. The method according to claim 4, characterized in that: The relative density is 1.12, the alcohol content is 40%, and the standing time is 24 hours.

8. The method according to claim 4, characterized in that: The medicine is in the form of granules, tablets, capsules or oral liquids, the auxiliary materials include sweeteners and excipients, the sweetener is sucralose, and the excipient is maltodextrin.

9. Use of the Chinese medicine according to any one of claims 1 to 3 in the preparation of a medicament for treating diseases caused by exogenous wind-heat or excessive heat in the lung and stomach, wherein the diseases include dry throat, sore throat, fever, yellow sputum, acute pharyngitis, acute laryngitis, acute tonsillitis or upper respiratory tract infection.

10. The use according to claim 9, characterized in that The drug has at least one of the following pharmacological effects: anti-inflammatory, analgesic, antipyretic, and antibacterial.

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