Traditional Chinese medicine composition for treating influenza and application thereof

Abstract

The invention provides a traditional Chinese medicine composition for treating influenza. The traditional Chinese medicine composition is prepared from the following medicinal components in parts by weight: 8-12 parts of radix puerariae, 1-5 parts of roasted herba ephedrae, 18-22 parts of gypsum, 3-7 parts of semen armeniacae amarae, 4-8 parts of radix bupleuri, 4-8 parts of notopterygium root, 4-8 parts of folium isatidis, 4-8 parts of medicated leaven and 1-5 parts of liquorice. The flu is exterior cold and interior heat syndrome. The traditional Chinese medicine composition is capable of alleviating drug properties, preventing excessive warm-dryness of ephedra and preventing cold-cold stomach injury of gypsum. The medicines are used for supplementing the deficiency of monarch and ministerial drugs, so that the effects of relieving exterior syndrome, dispelling cold and clearing internal heat are enhanced. According to the formula, all the medicines are combined to achieve the effects of relieving exterior syndrome, dispelling cold and clearing internal heat, so that evil is removed, exterior syndrome is eliminated, qi and blood are smooth, and the whole function is achieved. Generally, the prescription is rigorous, the compatibility is proper, and the traditional Chinese medicine is clinically effective.

Description

Technical Field

[0001] The invention belongs to the field of traditional Chinese medicines, and specifically provides a traditional Chinese medicine composition for treating influenza. Background Art

[0002] Influenza (flu), also known as seasonal cold in traditional Chinese medicine, is caused by epidemic pathogens, with sudden high fever, headache, body aches, fatigue, cough, sore throat, or abdominal pain, nausea, vomiting and diarrhea as the main clinical manifestations. Influenza infection in children can lead to death, and the risk of death in children with underlying diseases is significantly higher than that of healthy children. Therefore, early intervention of this disease, especially the use of traditional Chinese medicine intervention measures in the mild and moderate stages, is of great significance to the prognosis of the disease.

[0003] Influenza in children is mainly caused by influenza A virus and influenza B virus. Children have the special physiological characteristics of "excessive yang and insufficient yin". "Gezhi Yu Lun" once recorded: "Children before the age of sixteen are born with pure yang qi, which is hot". In previous studies, we found that influenza in children often occurs after the end of October. At this time, the indoor temperature in the north is generally high. Due to the improvement of living conditions, parents pay too much attention to their children's diet, feed them with rich and greasy food, even nourishing food, and often wear too warm clothes when going out, which can easily cause the children to have excess yang and excessive heat in the lungs and stomach. After the arrival of winter, the climate is cold, and the seasonal evils and epidemic toxins are often accompanied by wind and cold evils. After the children with yang-heat constitution are affected by the seasonal evils and epidemic toxins and wind and cold evils, they will turn into heat in a short time, forming a cold on the surface and heat on the inside, which is manifested as fever, aversion to cold, headache, red and swollen throat, red tongue, thin yellow fur, etc.

[0004] Influenza in children often has an acute onset, is highly contagious, and has a wide range of transmission. Once it becomes prevalent, the consequences are serious and the impact is huge. Western medicine should provide more vaccinations and antiviral treatments. However, since influenza viruses are prone to mutation, if you want to be protected by influenza vaccines, you need to get new vaccines every year. Oseltamivir has less drug resistance than other anti-influenza drugs. As a neuraminidase inhibitor, it has a clear target, has excellent anti-influenza effects, and can effectively reduce fever in a timely manner. However, children have different physical constitutions and their clinical symptoms are also different. Oseltamivir cannot work on the above specific symptoms. There are also adverse reactions represented by gastrointestinal symptoms. With the continuous development of history, the understanding of influenza in traditional Chinese medicine has become more and more perfect, and the treatment of influenza has gradually matured. The improvement of traditional Chinese medicine symptoms of influenza is particularly prominent, and there are fewer adverse reactions. Summary of the invention

[0005] In view of the deficiencies in the prior art, the present invention aims to provide a Chinese medicine composition for treating influenza with strong pertinence and clinical effectiveness, which is of great significance, especially for influenza A and influenza B in children, which is also called superficial cold and internal heat syndrome in traditional Chinese medicine.

[0006] The technical solution of the present invention is:

[0007] A traditional Chinese medicine composition for treating influenza, the drug components and dosages by weight are: 8-12 parts of kudzu root, 1-5 parts of ephedra, 18-22 parts of gypsum, 3-7 parts of apricot kernel, 4-8 parts of bupleurum, 4-8 parts of notopterygium, 4-8 parts of isatis indigotica, 4-8 parts of liushenqu and 1-5 parts of liquorice.

[0008] Preferably, the drug components and their dosages by weight are: 10 parts of Pueraria root, 3 parts of Ephedra, 20 parts of Gypsum, 5 parts of Apricot kernel, 6 parts of Bupleurum, 5 parts of Notopterygium wilfordii, 6 parts of Isatis indigotica, 6 parts of Liushenqu, and 3 parts of Licorice.

[0009] Prescription of the present invention is explained as follows:

[0010] This disease is mostly caused by wind-cold binding the surface and internal heat accumulation. This disease has many clinical syndromes, so the basic treatment principle for clinical treatment should be to dispel the cold and clear the lung heat. Pueraria root is sweet, pungent and cool. It enters the lung, spleen and stomach meridians; it dispels the surface evil, clears away muscle heat, and clears the yang to raise the body fluids and moisten the tendons and veins. Ephedra is pungent, slightly bitter and warm. It enters the lung and bladder meridians. It opens the closed skin and fur outside and restores the normal state of lung metal to clear and descend inside. The two are used together, so Pueraria root and ephedra are both monarch drugs. Gypsum tastes pungent and sweet. Pungent can disperse and sweet can regulate the middle, so it can clear lung heat and stop cough and asthma. When used with ephedra, one is cold and the other is warm. Ephedra gets the coldness of gypsum and slows down its warm and dry nature. Gypsum leads ephedra into the interior and slows down the sweating effect. Apricot kernel is bitter and slightly warm, so it can reduce and release lung qi, and when qi is reduced, phlegm will be eliminated and cough will be stopped. Paired with ephedra, ephedra promotes the flow of lung qi, while apricot kernel promotes and descends lung qi. The two herbs, one promotes and the other descends, complement each other and are in line with the lung's function. Qianghuo, pungent, bitter, and warm. It enters the lung and bladder meridians. This product has a strong medicinal property and can both disperse and release the exterior, and can also remove dampness and relieve pain. Bupleurum, pungent, bitter, and slightly cold. It enters the liver, gallbladder, spleen, and lung meridians, and can both disperse wind and cold and reduce fever. Isatis indigotica, bitter, and cold. It enters the heart and stomach meridians. It can enter the qi division to clear heat and detoxify, and enter the blood division to cool blood and relieve sore throat. Liushenqu, sweet, pungent, and warm. It enters the spleen and stomach meridians. This product can disperse and digest food, strengthen the spleen and stimulate appetite, and can also slightly relieve the exterior and reduce fever, so it is particularly suitable for exogenous symptoms and food stagnation. Licorice, sweet and neutral. It enters the heart, lung, spleen, and stomach meridians. This product moderates the medicinal property, preventing ephedra from being too warm and dry, and preventing gypsum from being too cold and damaging the stomach. The above medicines make up for the deficiency of the monarch and minister, and strengthen the efficacy of this prescription in relieving the exterior and dispersing cold and clearing the interior heat. This prescription is used in combination with other medicines to achieve the effect of relieving the exterior and dispersing cold, clearing the interior heat, so that the evil is removed from the exterior, and the qi and blood flow smoothly, thus achieving the full effect. Looking at the whole prescription, the formula is rigorous and the compatibility is appropriate, and it is definitely effective in clinical use.

[0011] The ratio of the formulation of the present invention is also very important, and it has been proven to be effective within the dosage range of the formulation of the present invention.

[0012] The preparation method of the Chinese medicine composition of the present invention comprises:

[0013] S1: Mix the Chinese medicinal materials uniformly according to the ratio of components and contents;

[0014] S2: Place the mixed Chinese medicinal materials in a casserole and soak them in water for 10-30 minutes, wherein the volume mass ratio of water to Chinese medicinal materials is 2-5 ml: 1 g;

[0015] S3: first boil over high heat, then simmer over low heat for 10-30 minutes, and the filtered medicinal liquid is the desired Chinese medicine composition.

[0016] Preferably, the preparation method further comprises:

[0017] S4: adding water to cover the API after filtering out the drug solution in step S3, and repeating step S3 to filter out the drug solution for the second time;

[0018] S5: Mix the medicinal liquid filtered out in step S3 with the second medicinal liquid filtered out in step S4 to obtain the desired Chinese medicine composition.

[0019] Preferably, the Chinese medicine composition is prepared into any one of the dosage forms such as tablets, granules, pills, capsules, decoctions and oral liquids.

[0020] The present invention also provides the use of the traditional Chinese medicine composition in preparing medicines for treating influenza or common cold.

[0021] The influenza is a syndrome of cold on the surface and heat on the inside, or the influenza is influenza A or influenza B. Preferably, the influenza A is HINI influenza A.

[0022] Preferably, the Chinese medicine composition is used in the preparation of medicines for treating influenza or colds in children.

[0023] More preferably, the child is a child aged 2-14 years old.

[0024] Compared with the prior art, the advantages of the present invention are:

[0025] 1. The clinical observation data of the present invention proves that the negative rate of nucleic acid PCR detection in the test group after 5 days of treatment with the Chinese medicine composition of the present invention is 89.83%, and the cure rate of traditional Chinese medicine syndrome is 91.52%, without the occurrence of complications, severe and critical diseases.

[0026] 2. The pharmacodynamic study data of the present invention prove that the Chinese medicine composition of the present invention has a good therapeutic effect, which can significantly reduce the viral load in the lung tissue of mice and significantly increase the sIgA level in the upper respiratory tract lavage fluid; significantly reduce the levels of TNF-α, IL-2, IL-6, and IFN-γ in the serum of mice; significantly reduce the lung index and lung tissue lesion scores of each group of mice; and significantly reduce the relative expression of TNFR, NF-κB, and JNK1 proteins in the lung tissue of mice.

[0027] 3. The repeated administration toxicity test and single administration toxicity test of the Chinese medicine composition of the present invention in rats by oral gavage for 3 months showed that the Chinese medicine composition of the present invention is highly safe and has no toxic side effects.

[0028] 4. The Chinese medicine composition of the present invention is mainly aimed at the syndrome of coldness in the exterior and heat in the interior. This Chinese medicine composition alleviates the medicinal properties, prevents ephedra from being too warm and dry, and prevents gypsum from being cold and damaging the stomach. The above medicines make up for the deficiencies of the monarch and minister, and strengthen the efficacy of this prescription in relieving the exterior and dispelling cold and clearing the interior heat. This prescription is used in combination with various medicines to play the role of relieving the exterior and dispelling cold, and clearing the interior heat, so that the evil is removed from the exterior, and the qi and blood are smooth and the whole work is collected. Looking at the whole prescription, the prescription is rigorous, the compatibility is appropriate, and it is clinically effective. Compared with the formula of the comparative example, the prescription and compatibility of the Chinese medicine composition of the present invention are more reasonable, and the curative effect is better, which proves that the synergistic effect is produced between the prescription drugs of the present invention.

[0029] The detailed structure of the present invention is further described below in conjunction with specific embodiments. The weight parts in the embodiments are dosage ratios and can be mass units such as "mg", "g" or "kg". DETAILED DESCRIPTION

[0030] Example 1

[0031] A traditional Chinese medicine composition for treating influenza, the drug components and dosages by weight of which are: 10 parts of kudzu root, 3 parts of roasted ephedra, 20 parts of gypsum, 5 parts of bitter almond, 6 parts of bupleurum, 5 parts of notopterygium, 6 parts of isatis indigotica, 6 parts of liushenqu and 3 parts of liquorice.

[0032] The preparation method is:

[0033] S1: Mix the Chinese medicinal materials uniformly according to the ratio of components and contents;

[0034] S2: Place the mixed Chinese medicinal materials in a casserole and soak them in water for 15 minutes, wherein the volume mass ratio of water to Chinese medicinal materials is 2 ml:1 g;

[0035] S3: first boil over high heat, then simmer over low heat for 25 min, and filter out the liquid medicine;

[0036] S4: adding water to cover the API after filtering out the drug solution in step S3, and repeating step S3 to filter out the drug solution for the second time;

[0037] S5: Mix the medicinal liquid filtered out in step S3 with the second medicinal liquid filtered out in step S4, and concentrate to 50 mL to obtain the desired Chinese medicine composition.

[0038] Comparative Example 1

[0039] A traditional Chinese medicine composition, the drug components and dosages by weight are: 3 parts of roasted ephedra, 20 parts of gypsum, 5 parts of bitter almonds, 6 parts of bupleurum, and 3 parts of liquorice.

[0040] The preparation method is the same as that in Example 1.

[0041] Comparative Example 2

[0042] A traditional Chinese medicine composition, the drug components and dosages by weight are: 10 parts of kudzu root, 6 parts of bupleurum, 5 parts of notopterygium, 6 parts of isatis indigotica, 6 parts of liushenqu and 3 parts of liquorice.

[0043] The preparation method is the same as that in Example 1.

[0044] Example 2

[0045] A traditional Chinese medicine composition for treating influenza, the drug components and dosages by weight of which are: 9 parts of kudzu root, 3 parts of roasted ephedra, 18 parts of gypsum, 4 parts of bitter almond, 5 parts of bupleurum, 4 parts of notopterygium, 5 parts of isatis indigotica, 5 parts of liushenqu and 3 parts of liquorice.

[0046] The preparation method is:

[0047] S1: Mix the Chinese medicinal materials uniformly according to the ratio of components and contents;

[0048] S2: Place the mixed Chinese medicinal materials in a casserole and soak them in water for 20 minutes, wherein the volume mass ratio of water to Chinese medicinal materials is 4 ml:1 g;

[0049] S3: first boil over high heat, then simmer over low heat for 20 min, and filter out the liquid medicine;

[0050] S4: adding water to cover the API after filtering out the drug solution in step S3, and repeating step S3 to filter out the drug solution for the second time;

[0051] S5: Mix the medicinal liquid filtered out in step S3 with the second medicinal liquid filtered out in step S4, and concentrate them to 50-60 mL to obtain the desired Chinese medicine composition.

[0052] Single-dose toxicity test of Example 1 by oral administration in rats

[0053] This study was carried out in accordance with the national GLP standard requirements, and the acute toxic reactions and deaths of SD rats after oral administration of the drug solution of Example 1 three times within 24 hours were observed, so as to provide reference data for repeated dosing toxicity tests.

[0054] The test selected 40 SPF-grade SD rats that passed the quarantine, half of which were male and half were female, and were raised in 475mm×350mm×200mm cages, with 5 rats in each cage. The animals were raised in accordance with the international (GB14925-2010) SPF-grade experimental animal environmental conditions, and the experimental animals were quarantined and adapted to the environment for 5 days. This experiment observed the acute toxicity test reaction of Example 1 by oral gavage in SD rats. 40 SD rats, half of which were male and half were female, and their body weight was 180.3-203.8g. They were randomly divided into 2 groups according to gender and body weight, namely, a blank control group and Example 1 group (63g decoction piece / kg), with 20 rats in each group. Before the experiment, the animals were fasted but not watered for more than 12 hours, and then 20 mL / kg of pure water and Example 1 were given by oral gavage, respectively. The animals were given 3 times on the day of administration (6 hours between two administrations), and the poisoning manifestations and characteristics, the occurrence and recovery time of toxic reactions, and the death of each group of experimental animals were closely observed and recorded within 0 to 4 hours after each administration. Then, the animals were observed twice a day, once in the morning and once in the afternoon, for 14 consecutive days. The experimental animals were weighed before administration on the day of administration and on the 4th, 7th, 10th, and 14th days after administration, and the weight changes and deaths of the experimental animals were recorded.

[0055] Test results: Effects on general activity, poisoning symptoms and death of experimental animals: Within 0 to 4 hours after each oral gavage, the rats in the blank control group and Example 1 group had no obvious abnormalities in their autonomous activity, mental state and diet, and no related toxic reactions or death of experimental animals were observed. After continuous observation for 14 days after administration, the rats in the blank control group and Example 1 group had no obvious abnormalities in their autonomous activity, mental state and diet, and no related toxic reactions or death of experimental animals were observed.

[0056] Effect on body weight: The experimental animals were weighed before administration on the day of administration, and on the 4th, 7th, 10th and 14th days after administration. There was no statistically significant difference in the body weight of the experimental animals between the Example 1 group and the blank control group during the same period, and the body weight growth was within the normal growth range, indicating that oral administration of Example 1 to SD rats had no significant effect on the body weight growth of the rats

[0057] At the end of the experiment, gross anatomy of SD rats was observed with the naked eye: no obvious abnormalities were found on the surface and cross-section of each organ.

[0058] Conclusion: Under the experimental conditions, rats were orally administered Example 1 at a volume of 20 mL / kg, three times a day (with a 6-hour interval between two administrations), and no related toxic reactions or deaths were observed in the experimental animals.

[0059] Table 1 Effect of Example 1 on the body weight of SD rats (n=20, )

[0060]

[0061] Example 1 Oral gavage toxicity test for three months in rats

[0062] Objective: This study was carried out in accordance with the national GLP standards to observe the toxic reactions of SD rats after oral administration of different doses of Example 1, to predict the possible clinical adverse reactions caused by Example 1, and to provide a reference for the monitoring and rescue measures of toxic reactions in clinical trials, clinical trials and clinical drug use.

[0063] Test method: 80 SD rats that passed the quarantine were selected, half male and half female, weighing 178.5-224.2g, 475×350×200mm 3 Cages were kept, 5 rats per cage. The animals were kept in accordance with the national standard (GB14925-2010) SPF-grade experimental animal environmental conditions, and the animals were quarantined and acclimatized for 7 days. The animals were randomly divided into 4 groups according to gender and weight, namely, a blank control group, low-, medium- and high-dose groups (10, 20, 40 g of slices / kg) of Example 1, 20 animals in each group, and each group was gavaged at a volume of 15 mL / kg for 3 consecutive months, and a 2-week drug withdrawal recovery period. At the end of the drug administration period (the 13th weekend), 40 rats were dissected as planned, and at the end of the recovery period (the 15th weekend), 40 rats were dissected as planned, half of them were male and half were female. The inspection items include: general clinical observation; body weight and food intake determination; hematological indicators, blood biochemistry, coagulation, and organ coefficient determination and inspection.

[0064] Test results:

[0065] General clinical observation: During the observation period of administration and recovery, the animals' appearance signs, behavioral activities, secretions and excretions of various cavities, and general conditions of the animals in each dose group of Example 1 were compared with the blank control group during the same period, and no abnormalities related to drug toxicity were found. During the test, all animals were euthanized as planned, and no animals died abnormally during the test.

[0066] Body weight: Compared with the blank control group during the same period, no abnormalities of toxicological significance were found in the body weights of the various dosage groups in Example 1.

[0067] Food intake: Compared with the blank control group during the same period, the average food intake of each dosage group in Example 1 showed no abnormality of toxicological significance.

[0068] Routine hematological examination: Compared with the blank control group during the same period, no abnormalities of toxicological significance were found in the hematological indicators of each dosage group in Example 1.

[0069] Blood biochemical examination: Compared with the blank control group during the same period, no abnormalities of toxicological significance were found in the blood biochemical indices of each dosage group in Example 1.

[0070] Coagulation test: compared with the blank control group during the same period, no abnormalities of toxicological significance were found in the coagulation indexes of each dosage group in Example 1.

[0071] Organ coefficients: Compared with the blank control group during the same period, no abnormalities of toxicological significance were found in the organ coefficients of each dosage group in Example 1.

[0072] Conclusion: Under the experimental conditions, no obvious toxic reaction was observed in SD rats after oral administration of Example 1 for three months.

[0073] Table 2 Effect of Example 1 on animal body weight (g, )

[0074]

[0075]

[0076] Note: Compared with the blank control group, there was no statistical difference in each dosage group of Example 1.

[0077] Table 3 Effect of Example 1 on the average food intake of animals (g / animal / day, )

[0078]

[0079] Note: Compared with the blank control group, there was no statistical difference in each dosage group of Example 1.

[0080] Table 4 Effect of Example 1 on Animal Hematology

[0081]

[0082] Note: Compared with the blank control group, * P<0.05.

[0083] Table 5 Effect of Example 1 on Animal Blood Biochemical Indexes

[0084]

[0085]

[0086] Note: Compared with the blank control group, * P<0.05.

[0087] Table 6 Effect of Example 1 on Animal Coagulation

[0088]

[0089] Note: Compared with the blank control group, there was no statistical difference in each dosage group of Example 1

[0090] Table 7 Effect of Example 1 on Animal Organ Coefficients ( mg / g)

[0091]

[0092]

[0093] Note: Compared with the blank control group, * P<0.05.

[0094] Pharmacodynamics study data of the formulation of Example 1

[0095] Materials and Methods

[0096] 1 Test materials

[0097] Test substance (test sample): Chinese medicine composition of Example 1 and Comparative Example 1 and Comparative Example 2, clinical intended dosage: 63g slices / day, clinical intended route: oral, 25g once (equivalent to 31.5g slices), twice a day, clinical intended course of treatment: 5 days, positive control group, name: Oseltamivir phosphate capsules, specification: 75mg / capsule, manufacturer: Hoff-Mer-Roche GmbH, Basel, Switzerland

[0098] The virus strain was influenza A virus mouse lung-adapted strain A / PR / 8 / 34 (H1N1) (abbreviated as PR8), provided by the Institute of Viral Diseases, Chinese Center for Disease Control and Prevention, and stored at -80°C for future use.

[0099] Experimental animals: 60 SPF-grade ICR female mice, weight range: 13-15g; (Hunan Slake Jingda Experimental Animal Co., Ltd.)

[0100] 2 Main reagents

[0101] Isoflurane (Hebei Yipin Pharmaceutical Co., Ltd.); influenza A virus nucleic acid detection kit (Shanghai Zhijiang Biotechnology Co., Ltd.); secretory immunoglobulin A (sIgA), tumor necrosis factor α (TNF-α), interferon γ (IFN-γ), interleukin 2 (IL-2), interleukin 6 (IL-6) ELISA detection kit (Wuhan Cloud Clone Technology Co., Ltd.); TRIzol reagent (Invitrogen); anti-human tumor necrosis factor receptor-1 (TNFR1) (Affinity), nuclear transcription factor-κB (NF-κB) (Affinity), Jun amino-terminal kinase (JNK) (Affinity), sodium pentobarbital (Shengxing Biotechnology Co., Ltd.); 4% paraformaldehyde (Wuhan Boster Bioengineering Co., Ltd.); HE staining solution set (Wuhan Sevier Biotechnology Co., Ltd.).

[0102] 3 Grouping and modeling

[0103] 60 mice were randomly divided into a blank group, a model group, Example 1, Comparative Examples 1 and 2, and a positive control group, with 10 mice in each group. After 1% pentobarbital anesthesia, 25 μL PBS was dripped into each nostril of each mouse in the blank group, and 25 μL virus solution was diluted and dripped into each nostril of each mouse in the remaining groups, with an infection dose of 20 half lethal (LD50).

[0104] 4. Dosage Method

[0105] Each group started to be medicated on the day of infection. The dosage of mice in Example 1, Comparative Examples 1 and Comparative Examples 2 was converted into a clinical equivalent dose for mice according to the body surface area = adult dosage × 0.0026 × 50 = 8.19 g crude drug / kg, and the dosage of mice in the oseltamivir phosphate group was converted into 27.5 mg / kg according to the adult clinical equivalent dose. The blank group and the model group were given distilled water, and each group of mice was gavaged at 0.2 mL / 10 g, once a day, for 3 consecutive days.

[0106] 5 Observation indicators and detection methods

[0107] ELISA method was used to detect the content of sIgA in upper respiratory tract lavage fluid and serum IFN-γ, TNF-α, IL-2, and IL-6. On the 9th day of the experiment, mice were anesthetized by intraperitoneal injection of 0.3% sodium pentobarbital (0.2 ml / 10 g), and blood was collected from the mouse eyeballs. The blood was placed in an EP tube and allowed to stand at 4°C for 30 minutes. The blood was centrifuged at 4°C and 3000 r / min (centrifugal radius 14 cm) for 15 minutes, and the serum was collected and stored at -80°C for testing. The mouse trachea was exposed by thoracotomy, and 1 ml of PBS buffer was injected into the mouse respiratory tract from the upper end of the trachea. The nasopharynx was repeatedly rinsed upward for 3 times, and the upper respiratory tract lavage fluid was collected. Subsequently, the blood was centrifuged at 4°C and 3000 r / min (centrifugal radius 14 cm) for 15 minutes, and the supernatant was collected and stored at -80°C for testing. Strictly follow the steps in the ELISA kit instructions to detect the levels of sIgA in upper respiratory tract lavage fluid and serum IFN-γ, TNF-α, IL-2, and IL-6.

[0108] RT-qPCR method for detecting viral load in lung tissue 15-20 mg of right lobe lung tissue of mice was taken, ground and centrifuged at 4°C, 3000r / min (centrifugal radius 14cm) for 15min, and the supernatant was collected as lung tissue homogenate. Total RNA was extracted using TRIzol and operated according to the instructions of the influenza A virus nucleic acid detection kit. PCR amplification was performed with target gene primers (parameters: 48°C 5min, 94°C 2min, 94°C 10s, 55°C 35s, 40 cycles), PR8 mRNA level was detected, and the copy number was calculated according to the standard curve using the scanning method.

[0109] HE staining to observe the pathological changes of lung tissue The left lobe lung tissue of mice was fixed in 4% paraformaldehyde, dehydrated by gradient alcohol, transparentized by xylene, embedded in paraffin, sliced, and stained with HE, and then read under an optical microscope. The lung tissue lesion score was comprehensively evaluated according to the degree of lung parenchymal lesions, interstitial lungs, and peribronchial inflammation. Healthy and lesion-free was scored as 0 points, mild interstitial pneumonia (accounting for less than 25% of the lung), moderate damage (accounting for 25% to 50% of the lung), severe damage (accounting for 51% to 75% of the lung), and very severe damage (accounting for more than 75% of the lung) were scored as 1, 2, 3, and 4 points, respectively.

[0110] The relative protein expression of TNFR1, NF-κB and JNK1 in lung tissue was determined by Western blotting, lung tissue protein extraction, BCA protein quantification, protein balancing, deformation, gel preparation, sample loading and electrophoresis, membrane transfer, membrane washing, blocking and primary antibody incubation, secondary antibody incubation, and chemiluminescence development.

[0111] 6 Data processing and statistical analysis

[0112] SPSS 16.0 software was used for statistical analysis, and the experimental data were expressed as mean ± standard deviation. Indicates that normality and variance homogeneity tests were performed when multiple groups were compared with homogeneity of variance. If normality was met (P>0.05), one-way analysis of variance (ANOVA) was used for statistical analysis, and LSD+Dunnet (homogeneous variance) or Tamhane's T2 (uneven variance) was selected for comparative analysis according to the variance homogeneity. If normality was not met (P<0.05), Kruskal-Wallis test was used, and Mann-Whitney test was used for pairwise comparison analysis. The statistical results were tested with α=0.05 as the test limit, where P<0.05 indicated statistical significance, and its biological significance was considered during evaluation.

[0113] Table 8 Comparison of viral load in lung tissue and sIgA level in upper respiratory tract lavage fluid of mice in each group ( n=10)

[0114]

[0115] Note: Compared with the blank control group, "**" indicates P < 0.01; compared with the model control group, Δ " indicates P<0.05, "ΔΔ" indicates P<0.01; compared with comparative example 1, "##" indicates P<0.01; compared with comparative example 2, "▲▲" indicates P<0.01

[0116] Compared with the blank control group, the viral load in the lung tissue of mice in the model group was significantly increased, and the sIgA level in the upper respiratory tract lavage fluid was significantly reduced (P < 0.01). Compared with the model group, the viral load in the lung tissue of mice in the comparative example 1 group, the comparative example 2 group, the embodiment 1 group and the positive control group was significantly reduced (P < 0.05 or P < 0.01), and the sIgA level in the upper respiratory tract lavage fluid was significantly increased (P < 0.05 or P < 0.01); compared with comparative examples 1 and 2, the viral load in the lung tissue of mice in embodiment 1 was significantly reduced, and the sIgA level in the upper respiratory tract lavage fluid was significantly increased, and there were statistical differences (P < 0.01); compared with the positive control group, there was no significant difference in embodiment 1, and there was no statistical difference (P> 0.05).

[0117] Experimental conclusion

[0118] The experimental results show that: under the experimental conditions, the viral load / copy number of Example 1, Comparative Example 1 and Comparative Example 2 groups and the positive control were significantly reduced (P<0.05 or P<0.01), and the sIgA in the upper respiratory tract lavage fluid was significantly increased (P<0.05 or P<0.01), indicating that Example 1, Comparative Example 1 and Comparative Example 2 have a significant improvement effect on the viral load in the lung tissue of mice and the sIgA level in the upper respiratory tract lavage fluid, and compared with Comparative Example 1 and Comparative Example 2, Example 1 has a better effect.

[0119] Table 9 Comparison of serum TNF-α, IL-2, IL-6, and IFN-γ levels in each group of mice ( n=10)

[0120]

[0121] Note: Compared with the blank control group, "**" indicates P<0.01; compared with the model control group, "Δ" indicates P<0.05, "ΔΔ" indicates P<0.01; compared with comparative example 1, "#" indicates P<0.05, "##" indicates P<0.01; compared with comparative example 2, "▲" indicates P<0.05, "▲▲" indicates P<0.01 Compared with the blank control group, the serum TNF-α, IL-2, IL-6, and IFN-γ levels of mice in the model group were significantly increased (P<0.01). Compared with the model group, the serum TNF-α, IL-2, IL-6, and IFN-γ levels of mice in comparative example 1, comparative example 2, example 1, and positive control group were significantly reduced (P<0.05 or P<0.01); compared with comparative example 1 and comparative example 2, example 1 had statistical differences (P<0.05 or P<0.01); compared with the positive control group, there was no significant difference in example 1, and there was no statistical difference (P>0.05).

[0122] Experimental conclusion

[0123] The experimental results show that: under the experimental conditions, in Example 1, Comparative Examples 1 and 2, and the positive control, the serum TNF-α, IL-2, IL-6, and IFN-γ levels of mice in each group were significantly reduced (P<0.05 or P<0.01), indicating that Example 1, Comparative Examples 1 and 2 have a significant improvement effect on the serum TNF-α, IL-2, IL-6, and IFN-γ content indicators of mice, and compared with Comparative Examples 1 and 2, Example 1 has a better effect.

[0124] Table 10 Comparison of lung index and lung tissue lesion scores of mice in each group ( n=10)

[0125]

[0126]

[0127] Note: Compared with the blank control group, "**" indicates P < 0.01; compared with the model control group, "Δ" indicates P < 0.05, "ΔΔ" indicates P < 0.01; compared with comparative example 1, "##" indicates P < 0.01; compared with comparative example 2, "▲▲" indicates P < 0.01

[0128] Compared with the blank control group, the lung index and lung tissue lesion score of mice in the model group were significantly increased (P < 0.01). Compared with the model group, the lung index and lung tissue lesion score of mice in the comparative example 1 group, the comparative example 2 group, the embodiment 1 group and the positive control group were significantly reduced (P < 0.05 or P < 0.01); compared with the comparative example 1 and the comparative example 2, there was a statistical difference in embodiment 1 (P < 0.05 or P < 0.01); compared with the positive control group, there was no significant difference in embodiment 1, and there was no statistical difference (P> 0.05).

[0129] Experimental conclusion

[0130] The experimental results show that: under the experimental conditions, the lung index and lung tissue lesion score of mice in Example 1, Comparative Examples 1 and 2 and the positive control were significantly reduced (P<0.05 or P<0.01), indicating that Example 1, Comparative Examples 1 and 2 have a significant improvement effect on the lung index and lung tissue lesion score of mice, and compared with Comparative Examples 1 and 2, Example 1 has a better effect.

[0131] Table 11 Comparison of relative expression levels of TNFR, NF-κB and JNK1 proteins in lung tissues of mice in each group ( n=10)

[0132]

[0133] Note: Compared with the blank control group, "**" indicates P < 0.01; compared with the model control group, "Δ" indicates P < 0.05, "ΔΔ" indicates P < 0.01; compared with comparative example 1, "#" indicates P < 0.05; compared with comparative example 2, "▲" indicates P < 0.05

[0134] Compared with the blank control group, the relative expression of TNFR, NF-κB, and JNK1 proteins in the lung tissue of the model group mice was significantly increased (P < 0.01). Compared with the model group, the relative expression of TNFR, NF-κB, and JNK1 proteins in the lung tissue of the comparative example 1 group, the comparative example 2 group, the example 1 group, and the positive control group was significantly reduced (P < 0.05 or P < 0.01); compared with the comparative example 1 and the comparative example 2, there was a statistical difference in example 1 (P < 0.05); compared with the positive control group, there was no significant difference in example 1, and there was no statistical difference (P> 0.05).

[0135] Experimental conclusion

[0136] The experimental results show that: under the experimental conditions, the relative expression levels of TNFR, NF-κB, and JNK1 proteins in mouse lung tissue in Example 1, Comparative Example 1, and Comparative Example 2 groups and the positive control were significantly reduced (P<0.05 or P<0.01), indicating that Example 1, Comparative Example 1, and Comparative Example 2 have a significant improvement effect on the relative expression levels of TNFR, NF-κB, and JNK1 proteins in mouse serum and mouse lung tissue, and compared with Comparative Example 1 and Comparative Example 2, Example 1 has a better effect.

[0137] Clinical observation data

[0138] Test title: Clinical observation on the treatment of influenza (exterior cold and interior heat syndrome) in children with "Xiao'er Qingjie Fang" (the Chinese medicine composition of the present invention).

[0139] Purpose of the experiment: To evaluate the effectiveness and safety of "Xiao'er Qingjie Fang" (the Chinese medicine composition of the present invention) in treating influenza (syndrome of cold on the surface and heat inside) in children.

[0140] General data: This study selected 120 children with influenza as research subjects and randomly divided them into a control group and an observation group, 60 cases in each group. The observation group included 32 males and 28 females, aged 2 to 13 years old, with an average age of (5.67±1.63) years old; the course of disease was 1 to 5 days, with an average of (3.02±1.03) days. The control group included 33 males and 27 females, aged 2 to 14 years old, with an average age of (5.58±1.90) years old; the course of disease was 1 to 6 days, with an average of (3.15±1.05) days. The general data of the two groups were compared, and there was no significant difference between the two groups (P>0.05), which was comparable.

[0141] Diagnostic criteria

[0142] 1. Western medicine diagnostic criteria

[0143] Refer to the clinical diagnostic criteria for influenza in children in the Expert Consensus on Diagnosis and Treatment of Influenza in Children (2020 Edition). The diagnosis of influenza in children is mainly based on epidemiological history, clinical manifestations and etiological examination.

[0144] (1) Influenza-like illness: During the influenza epidemic season, the following symptoms occur: ① Fever, body temperature ≥ 38°C; ② Accompanied by cough and / or sore throat.

[0145] (2) Clinically diagnosed cases: patients with the above-mentioned clinical manifestations of influenza and an epidemiological history (close contact with suspected or confirmed influenza patients without effective personal protection within 7 days before onset of illness, or being one of the participants in a cluster of influenza-like cases, or having clear evidence of infection to others) and other diseases that cause influenza-like symptoms are excluded.

[0146] (3) Confirmed cases: Clinically diagnosed influenza cases with one or more of the following positive etiological test results: ① positive influenza virus nucleic acid test; ② positive influenza antigen test; ③ positive influenza virus isolation and culture; ④ influenza virus-specific IgG antibody levels in both acute and convalescent serum samples increased by 4 times or more.

[0147] (4) Severe cases: Influenza cases are classified as severe cases if they present with one or more of the following: ① Difficulty breathing and / or increased respiratory rate: >30 times / min for children over 5 years old; >40 times / min for 1-5 years old; >50 times / min for 2-12 months old; >60 times / min for newborns up to 2 months old; ② Mental changes: slow reaction, lethargy, restlessness, convulsions, etc.; ③ Severe vomiting, diarrhea, and signs of dehydration; ④ Oliguria: urine volume <0.8 mL / (kg·h) for children, or daily urine volume <200 mL / m for infants 2 , preschool children <300 mL / m 2 , school-age children <400 mL / m 2 , children over 14 years old <17mL / h, or acute renal failure occurs; ⑤ Pneumonia; ⑥ Significant aggravation of the original underlying disease; ⑦ Other clinical conditions requiring hospitalization.

[0148] (5) Critical cases: patients with any of the following conditions: ① respiratory failure; ② acute necrotizing encephalopathy (ANE); ③ septic shock; ④ multiple organ failure; ⑤ other serious clinical conditions requiring intensive care.

[0149] (II) TCM syndrome differentiation criteria (syndrome of cold on the surface and heat inside)

[0150] Refer to the "Influenza Diagnosis and Treatment Guidelines 2020 Edition"

[0151] Main symptoms: ① fever, ② chills (or clear runny nose);

[0152] Secondary symptoms: ① Headache, ② Body aches, ③ Sore throat, ④ Nasal congestion, ⑤ Clear runny nose, ⑥ Thirst.

[0153] Tongue and pulse: red tongue, thin or yellow coating, rapid pulse.

[0154] The child must meet all the main symptom requirements and meet 2 or more secondary symptom requirements. Combined with the tongue and pulse, the syndrome of superficial cold and internal heat can be diagnosed.

[0155] 1. Criteria for inclusion of cases

[0156] (1) Meet the clinical diagnostic criteria for influenza in children in the "Expert Consensus on Diagnosis and Treatment of Influenza in Children (2020 Edition)";

[0157] (2) Meet the TCM criteria for pediatric influenza with cold on the surface and heat inside;

[0158] (3) A rapid viral antigen test result of a nasopharyngeal swab or oropharyngeal swab is positive, or an influenza virus nucleic acid test is positive within 24 hours;

[0159] (4) body temperature ≤ 38°C (axillary temperature) < 39.5°C at the time of consultation;

[0160] (5) The duration of illness was ≤ 48 hours (the duration of illness was defined as the time from onset to the current visit, and the onset time referred to the time when any one of the 10 influenza symptoms and signs, including fever, headache, muscle and joint pain, chills, shivering, fatigue, cough, sore throat, nasal congestion, and runny nose, appeared);

[0161] (6) Aged between 2 and 13 years old (<14 years old).

[0162] (II) Exclusion criteria

[0163] (1) Allergy to the experimental drug or its components or allergic constitution (allergy to two or more foods or drugs);

[0164] (2) Suffering from pharyngitis, herpetic pharyngitis, suppurative tonsillitis, etc.;

[0165] (3) Influenza complications such as sinusitis, otitis media, bronchitis, pneumonia, etc. have occurred;

[0166] (4) severe or critical cases of influenza;

[0167] (5) Children with spleen and stomach deficiency and coldness;

[0168] (6) The total white blood cell count is greater than the upper limit of the reference value (ULN), and the researchers determine that the child is suspected of having bacterial infection;

[0169] (7) Use of antiviral drugs, or Chinese or Western medicines for the treatment of influenza within 48 hours before this visit;

[0170] (8) Currently receiving systemic steroid therapy or other immunosuppressive therapy;

[0171] (9) History of epilepsy or febrile convulsions, or recurrent respiratory tract infections;

[0172] (10) Patients with severe malnutrition, rickets, and severe primary diseases of the heart, brain, liver, kidney, and hematopoietic systems;

[0173] The researcher decided to withdraw

[0174] 1) Serious adverse events occur and the trial should be stopped according to the doctor's judgment;

[0175] 2) If the child's body temperature (axillary temperature) is greater than 38.5°C, the researcher may add antipyretic and analgesic drugs (such as acetaminophen, ibuprofen) according to the situation to protect the subject. If the high fever persists, such as the fever symptoms are not relieved for more than 24 hours, it is recommended that the subject withdraw from the trial and be treated as an ineffective case;

[0176] 3) During the trial, the subjects have other diseases that affect the judgment of efficacy and safety;

[0177] 4) Subjects with poor compliance (compliance with trial medication <80%, or >120%), or who automatically change medications midway or add Chinese and Western medicines prohibited by this protocol;

[0178] 5) After taking the medicine, if the subject's condition worsens or complications occur, such as pneumonia, the medicine should be stopped and effective treatment measures should be taken. The subject should complete various laboratory tests, withdraw from the trial, and be treated as an ineffective case;

[0179] 6) Patients who are found to have seriously violated the inclusion or exclusion criteria, or who have not taken medication after inclusion despite meeting the inclusion criteria, or who have no follow-up records;

[0180] 7) Those whose viral nucleic acid test result is negative by RT-PCR method.

[0181] Subjects voluntarily withdraw

[0182] 1) For whatever reason, the subject is unwilling or unable to continue the clinical trial and requests to withdraw from the trial to the attending physician, thereby terminating the trial;

[0183] 2) Subjects who did not explicitly request to withdraw from the trial but no longer accepted medication and testing and were lost to follow-up.

[0184] The completion of the planned follow-up of the last case marks the end of a clinical trial.

[0185] Trial Drugs

[0186] The Xiaoer Qingjie Recipe (the Chinese medicine composition of the present invention) is composed of 9 herbs: Pueraria root, roasted ephedra, gypsum, bitter almond, bupleurum, notopterygium root, isatis indigotica, Liushenqu and liquorice.

[0187] Table 12 Medicinal material standards and prescription quantities (g)

[0188]

[0189] The preparation method is as follows: prepare according to the method of Example 1 to obtain the Chinese medicine composition decoction of the present invention.

[0190] Positive control group, name: Oseltamivir phosphate granules, specification: 15 mg (calculated as oseltamivir), production unit: Yichang East Sunshine Changjiang Pharmaceutical Co., Ltd.

[0191] Treatment options

[0192] 1. Usage and dosage of the Chinese medicine composition of the present invention: According to the Chinese medicine decoction, for children under 6 years old (2-5 years old), 1 / 2 of the original prescription amount is administered twice a day for 5 consecutive days. For children 6 years old and above (6-13 years old), the original prescription amount is administered twice a day for 5 consecutive days.

[0193] Control group medication:

[0194] For patients with body weight less than or equal to 15 kg, take 30 mg each time, twice a day for 5 consecutive days;

[0195] For patients weighing less than or equal to 15 kg and greater than or equal to 23 kg, take 45 mg each time, twice a day for 5 consecutive days;

[0196] For patients with a body weight of 23 kg or less and 40 kg or more, take 60 mg twice a day for 5 consecutive days.

[0197] For patients with body weight greater than or equal to 40 kg, take 75 mg each time, twice a day for 5 consecutive days.

[0198] 2. Research cycle

[0199] Treatment period (5 days) + follow-up period (2 days).

[0200] 3. Provisions on combined medication

[0201] (1) If the child's body temperature (axillary temperature) is >38.5℃, the researcher may add antipyretic and analgesic drugs (such as acetaminophen, ibuprofen) according to the situation to protect the subject. If the high fever persists, such as the fever symptoms are not relieved for more than 24 hours, the subject is advised to withdraw from the trial.

[0202] (2) Except for the experimental drugs and the prescribed emergency drugs, the use of other Chinese and Western medicines with similar efficacy or for the treatment of influenza, as well as other treatments related to the treatment of this disease (such as massage, acupuncture, etc.) is prohibited during the observation period;

[0203] (3) All concomitant medications of the subjects should be recorded in the research medical records, including the generic name, time of use, dosage, and reason for use, and a determination should be made as to whether they affect the efficacy of the trial medication.

[0204] Safety evaluation

[0205] Safety evaluation indicators and observation time points:

[0206] (1) Possible clinical adverse events / adverse reactions should be observed at any time after taking the medicine.

[0207] (2) Vital signs, such as blood pressure, respiration, body temperature (axillary temperature), heart rate, etc., are measured before and after treatment.

[0208] (3) Laboratory examination: routine blood test (WBC, RBC, HGB, PLT, NEUT, LYM), routine urine test (WBC, RBC, PH, PRO, KET, GLU), liver function test (ALT, AST, TBIL, ALP, γ-GGT), and renal function test (Cr, Urea / BUN);

[0209] (4) 12-lead electrocardiogram.

[0210] The incidence of clinical adverse events / adverse reactions was the main safety evaluation indicator.

[0211] Filter indicators

[0212] C-reactive protein (CRP), chest CT / X-ray. Whether to conduct the examination is determined by the researcher.

[0213] Diagnostic indicators

[0214] Throat swab test

[0215] 1) The first step for the enrollment is to select the rapid antigen detection method (colloidal gold method) for screening;

[0216] 2) For confirmed diagnosis, choose viral nucleic acid testing method.

[0217] Effectiveness evaluation

[0218] Observation indicators

[0219] (1) Demographic indicators: gender, age, height, weight, etc.

[0220] (2) General clinical information: personal history, medical history, physical examination, comorbidities and medications, start date of the trial, etc.

[0221] (3) Efficacy indicators:

[0222] ① Complete fever reduction time (hours). After the first treatment, the axillary temperature was measured every 6 hours and evaluated after 5 days of treatment.

[0223] ② Clinical recovery time (days): After the first treatment, CARIFS score was performed every 24 hours and evaluated after 5 days of treatment;

[0224] ③ The efficacy of TCM syndrome (cure rate) was scored once at baseline and 5 days after treatment, and evaluated after 5 days of treatment.

[0225] ④ The incidence of complications / severe / critical influenza was calculated and evaluated based on the occurrence of complications after 5 days of treatment;

[0226] ⑤ Virus conversion rate: influenza virus testing by throat swab at baseline and 5 days after treatment, and evaluation after 5 days of treatment.

[0227] Efficacy evaluation criteria

[0228] ① Definition of complete fever reduction: After treatment, the axillary temperature drops to 37.2℃ or below and is maintained for more than 24 hours.

[0229] ② Definition of clinical recovery: After treatment, fever completely subsides, and the score of the "symptom dimension" of each item in CARIFS is 0 or 1 point, and it lasts for more than 24 hours.

[0230] ③Definition of influenza complications: including sinusitis, otitis media, bronchitis and pneumonia, or hospitalization due to influenza.

[0231] ④ TCM syndrome efficacy evaluation criteria

[0232] Clinical recovery: symptoms basically disappear, and the TCM syndrome score decrease rate is ≥95%;

[0233] Markedly effective: symptoms were significantly improved, with a decrease rate of TCM syndrome score of ≥70% and <95%;

[0234] Effective: Symptoms are improved, and the TCM syndrome score decreases by ≥30% and <70%;

[0235] Ineffective: No improvement or aggravation of symptoms, and the TCM syndrome score reduction rate is <30%.

[0236] Note: The calculation formula (nimodipine method) is: [(score before treatment - score after treatment) / score before treatment] × 100%.

[0237] Quantitative Grading Standard of TCM Syndrome

[0238] It is formulated with reference to the "Technical Guidelines for the Design and Evaluation of Clinical Trials of Traditional Chinese Medicines for Common Pediatric Diseases - Influenza" and the "Guidelines for Clinical Research of New Traditional Chinese Medicines (Trial)".

[0239] Table 13 Quantitative standard for grading symptoms of TCM syndrome of cold on the surface and heat on the inside

[0240]

[0241] Note: Symptoms that cannot be clearly expressed due to age restrictions may not be evaluated.

[0242] Statistical analysis

[0243] 1. Definition of Dataset

[0244] Full Analysis Set (FAS): including all subjects who were randomly enrolled, took the drug at least once, and had at least one visit record. The full analysis data set was used for intention-to-treat (ITT) analysis. For missing values ​​of the primary efficacy evaluation indicators, the method of carrying the most recent observation data to the final result of the trial (LOCF method) was used.

[0245] Per-protocol set (PPS): includes all subjects who comply with the trial protocol, have no missing baseline variables, can measure the main variables, and have no major violations of the trial protocol.

[0246] Safety Set (SS): actual data of patients who have received at least one treatment and have safety indicators recorded. Data of patients who have withdrawn from treatment will not be transferred.

[0247] The FAS and PPS data sets were used for efficacy evaluation, and the SS data set was used for safety evaluation.

[0248] 2. Handling of missing values

[0249] For missing values ​​of efficacy indicators, the method of carrying forward the most recent observation data to the final results of the trial (LOCF method) is adopted. For missing values ​​of safety indicators, no carry-over is made.

[0250] 3. General Principles of Statistics

[0251] Quantitative data, describe the number of cases, mean, standard deviation, minimum, median, maximum, upper quartile (Q1), lower quartile (Q3), 95% confidence interval (95% CI). The comparison between treatment groups was performed using t-test. If the influence of covariates is considered, covariance analysis was used.

[0252] For time series data, the median survival time was described and the Kaplan-Meier curve was drawn. The Log-rank test was used to compare the difference in median survival time between the two groups. If the influence of the center or other factors was taken into account, Cox regression analysis was used.

[0253] Qualitative data described the number of cases and percentages of each category. The chi-square test and Fisher's exact probability method were used to compare the count data between treatment groups; the Wilcoxon rank sum test was used to compare the rank data between treatment groups. If the influence of the center or other factors was taken into account, the CMH chi-square test or logistic regression was used.

[0254] SAS v9.4 software was used for statistical analysis. Two-sided test was used, α = 0.05, and the confidence level of all confidence intervals was 95%.

[0255] 4. Multiplicity Adjustment

[0256] This study was a phase II dose-finding study and multiplicity adjustment was not considered.

[0257] For details of the specific analysis plan, please see the Statistical Analysis Plan.

[0258] Statistical results

[0259] 1. Study enrollment

[0260] A total of 120 subjects were enrolled in this trial, with 60 subjects in the test group and 60 subjects in the control group. 118 subjects entered FAS, including 59 subjects in the test group and 59 subjects in the control group; 104 subjects entered PPS, including 52 subjects in the test group and 52 subjects in the control group; 118 subjects entered SS, including 59 subjects in the test group and 59 subjects in the control group.

[0261] There was no statistically significant difference in the number of PPS subjects included in each group.

[0262] (II) Baseline comparability analysis: FAS and PPS analysis

[0263] For all subjects included in the FAS and PPS analysis, except for the PPS analysis of height and weight, there were no statistically significant differences in the baseline demographic data (sex, ethnicity, age group, age, height, weight) and disease-related conditions (current course of disease, nucleic acid PCR test results, pre-treatment comorbidities, past medical history, vaccination history, allergy history, pre-treatment comorbidities) between the groups, and they were comparable. Among them, the number of positive cases (%) of nucleic acid PCR test was 53 (89.83%) in the test group and 54 (91.53%) in the control group during FAS analysis, and 52 (100.0%) in the test group and 52 (100.0%) in the control group during PPS analysis. The current course of disease of the enrolled subjects was ≤24 hours.

[0264] There was no significant difference in the efficacy-related indicators, the total score of TCM syndrome and the single symptom score at baseline between the two groups. The conclusions of FAS and PPS were consistent.

[0265] (III) Compliance evaluation: FAS and PPS analysis

[0266] The FAS (PPS) analysis of compliance (%) showed that the test group was 95.208±8.257 (95.877±7.439) and the control group was 94.003±10.171 (95.952±8.048). There was no statistically significant difference between the two groups, and the conclusions of FAS and PPS were consistent.

[0267] The FAS (PPS) analysis of compliance (grade) showed no statistically significant difference. The conclusions of FAS and PPS were consistent.

[0268] (IV) Effectiveness evaluation: FAS and PPS analysis

[0269] 1. Flu symptom relief time (h)

[0270] FAS (PPS) analysis of the time of influenza symptom relief in the two groups showed that the median time of influenza symptom relief in the test group was 48 hours and that in the control group was 80 hours. The Log-Rank comparison between the two groups showed that the differences were statistically significant. The conclusions of FAS and PPS were consistent.

[0271] Table 14 Comparison of influenza symptom relief time between groups

[0272]

[0273] 2. Complete fever reduction time (hours)

[0274] The FAS (PPS) analysis of the time to complete fever reduction in the two groups showed that the median time for the test group was 48 hours and that for the control group was 80 hours. The time to complete fever reduction in the test group was less than that in the control group. The Log-Rank comparison between the two groups showed that the differences were statistically significant. The conclusions of FAS and PPS were consistent.

[0275] Table 15 Comparison of complete cooling time

[0276]

[0277] 3. TCM syndrome efficacy

[0278] The results of the FAS analysis of the covariance analysis of the TCM syndrome score at the end of the trial, excluding the interaction between the center and the group, and taking the TCM syndrome score at the baseline as the covariate, showed that there was no statistically significant difference between the groups in the decrease of the TCM syndrome score at the end of the trial compared with the baseline. Among them, the FAS analysis of the decrease of the TCM syndrome score at the end of the trial compared with the baseline was 9.3119 points in the trial group and 9.0953 points in the control group (P=0.3497).

[0279] According to FAS statistics, the cure rates of TCM syndrome at the end point (treatment day 05) of the trial group and the control group were 91.52% and 88.14%, respectively, and there was no significant difference between the two groups (P=0.4429).

[0280] 4. Disappearance time and disappearance rate of single symptom

[0281] According to the FAS set statistics, the disappearance time of fever, headache and body pain, red and swollen throat, runny nose, cough, fatigue and thirst was shorter in the experimental group than in the control group, and the difference was statistically significant (P < 0.05). The disappearance rate of the above single symptoms was higher in the experimental group than in the control group, and the difference was statistically significant (P < 0.05).

[0282] 5. Efficacy results - incidence of complications, severe and critical illnesses

[0283] Table 16 Statistics of the incidence of complications, severe and critical illnesses

[0284]

[0285] No complications, severe or critical illness occurred on treatment day 03, treatment day 05 and during the follow-up period.

[0286] 6. Virus negative conversion rate

[0287] Table 17 Comparison of influenza virus antigen detection in each group after treatment (FAS)

[0288]

[0289] Note: 1, positive-negative; 2, positive-positive; 3, positive-unchecked; 4, negative-negative; 5, negative-positive; 6, negative-unchecked.

[0290] According to the FAS set analysis, the influenza virus antigen detection kit (Shanghai Zhijiang Biotechnology Co., Ltd.) was used to detect influenza A and B viruses. The nucleic acid antigen negative rate on day 5 of treatment was 89.83% in the test group and 79.66% in the control group. There was no significant difference between the two groups (P=0.1725). According to the FAS set analysis, the influenza virus antigen negative conversion rate before and after treatment was 81.36% in the test group and 76.27% in the control group (P=0.4223), showing a trend of the test group>control group, but the comparison between the two groups did not indicate a significant difference.

[0291] 7. Safety evaluation: No obvious adverse events occurred during the trial, and the trial was safe and reliable.

[0292] In summary, in terms of the time for relief of influenza symptoms and the time for complete fever reduction, the test group was less than the control group, and the differences between the groups were statistically significant.

[0293] The above is a specific implementation mode of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field, within the technical scope disclosed by the present invention, can make equivalent replacements or changes according to the technical solutions and concepts of the present invention, which should be covered by the protection scope of the claims of the present invention.

Claims

1. A Chinese medicine composition for treating influenza, characterized in that: The drug components and dosage by weight are: 8-12 parts of kudzu root, 1-5 parts of ephedra, 18-22 parts of gypsum, 3-7 parts of apricot kernel, 4-8 parts of bupleurum, 4-8 parts of notopterygium, 4-8 parts of isatis indigotica, 4-8 parts of liushenqu and 1-5 parts of liquorice.

2. The Chinese medicine composition for treating influenza according to claim 1, characterized in that: The drug components and dosages by weight are: 10 parts of kudzu root, 3 parts of ephedra, 20 parts of gypsum, 5 parts of apricot kernel, 6 parts of bupleurum, 5 parts of notopterygium, 6 parts of isatis indigotica, 6 parts of liushenqu and 3 parts of liquorice.

3. The Chinese medicine composition for treating influenza according to claim 1 or 2, characterized in that: The preparation method comprises: S1: Mix the Chinese medicinal materials uniformly according to the proportion of components and contents; S2: Place the mixed Chinese medicine raw materials in a casserole and soak in water for 10-30min, where the volume mass ratio of water to Chinese medicine raw materials is 2-5ml:1g; S3: first boil over high heat, then simmer over low heat for 10-30 minutes, and the filtered medicinal liquid is the desired Chinese medicine composition.

4. The Chinese medicine composition for treating influenza according to claim 1 or 2, characterized in that: The preparation method also includes: S4: add water to cover the bulk drug after filtering out the liquid in step S3, repeat step S3 to filter out the liquid for the second time; S5: Mix the medicinal liquid filtered out in step S3 with the second medicinal liquid filtered out in step S4 to obtain the desired Chinese medicine composition.

5. The Chinese medicine composition for treating influenza according to claim 1 or 2, characterized in that: The traditional Chinese medicine composition is prepared into any one of tablets, granules, pills, capsules, decoctions and oral liquids.

6. Use of the Chinese medicine composition according to any one of claims 1 to 5 in the preparation of medicines for treating influenza or common cold.

7. The use according to claim 6, wherein the influenza is a syndrome of cold on the surface and heat on the inside, or the influenza is influenza A or influenza B.

8. The use according to claim 6, wherein the Chinese medicine composition is used in the preparation of medicines for treating influenza or common colds in children.

9. The use according to claim 8, wherein the child is a child aged 2-14 years old.

Images