A traditional Chinese medicine composition for treating influenza and its application

Abstract

The present invention provides a traditional Chinese medicine composition for treating influenza, comprising the following medicinal ingredients and dosages by weight: 8-12 parts of Puerariae Radix, 1-5 parts of roasted Ephedrae Herba, 18-22 parts of Gypsum Fibrosum, 3-7 parts of Apricot Kernel, 4-8 parts of Bupleurum Root, 4-8 parts of Notopterygium Incisum, 4-8 parts of Isatis Indigotica Folium, 4-8 parts of Liushenqu, and 1-5 parts of Licorice Root. The influenza presents as a syndrome of exterior cold and interior heat. This traditional Chinese medicine composition mitigates the effects of the medicinal properties, preventing the excessive warming and drying effects of Ephedrae Herba and the cooling and stomach-damaging effects of Gypsum Fibrosum. These herbs supplement the deficiencies of the monarch and ministerial herbs, enhancing the efficacy of this prescription in dispelling exterior cold and clearing interior heat. This prescription, when used in combination with these herbs, achieves the effects of dispelling exterior cold and clearing interior heat, dispelling exterior pathogens, and promoting the smooth flow of Qi and blood, ultimately achieving the desired effect. The entire prescription demonstrates rigorous formulation, appropriate compatibility, and clinical effectiveness.

Description

Technical Field

[0001] The present invention belongs to the field of traditional Chinese medicine, 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 (TCM), is caused by an epidemic pathogen. Its primary clinical manifestations include sudden onset of high fever, headache, body aches, fatigue, cough, sore throat, or abdominal pain, nausea, vomiting, and diarrhea. Influenza infection in children can be fatal, and the risk of death is significantly higher in children with underlying medical conditions than in healthy children. Therefore, early intervention, particularly Traditional Chinese Medicine interventions during the mild to moderate stages of the disease, is crucial for prognosis.

[0003] Influenza in children is primarily caused by influenza A and influenza B viruses. Children possess a unique physiological characteristic characterized by "constant excess of yang and deficiency of yin." The "Gezhi Yu Lun" (A Treatise on the Study of the Gezhi) states, "Under the age of sixteen, children possess a pure yang qi, which results in a high fever." Previous research has found that childhood influenza is most common after the end of October, when indoor temperatures in northern China are generally higher. Furthermore, with improved living conditions, parents are overly concerned with their children's diet, feeding them rich, greasy foods, even tonic foods, and often overdressing them when going out. This can lead to excess yang and excessive lung and stomach heat in children. With the arrival of winter, the cold weather often brings with it wind-cold pathogens. Children with a yang-heat constitution, when exposed to these pathogens, quickly experience internal heat, resulting in a syndrome of superficial cold and internal heat, characterized by fever, aversion to cold, headache, red and swollen throat, red tongue, and a thin, yellow coating.

[0004] Influenza in children often presents with an acute onset, is highly contagious, and spreads widely. Once an epidemic occurs, the consequences are severe and the impact is enormous. Western medicine should focus on vaccination and antiviral treatment. However, because influenza viruses are prone to mutation, protection through influenza vaccines requires annual vaccinations. Oseltamivir has a lower resistance rate than other anti-influenza drugs. As a neuraminidase inhibitor, it has a clear target, exhibits excellent anti-influenza effects, and can effectively reduce fever in a timely manner. However, children have different constitutions and exhibit varying clinical symptoms. Oseltamivir is not effective for these specific symptoms, and 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 increasingly refined, and the treatment of influenza has gradually matured. Traditional Chinese medicine is particularly effective in improving influenza symptoms and has fewer adverse reactions. Summary of the Invention

[0005] In view of the shortcomings of the existing technology, the purpose of the present invention is to provide a highly targeted and clinically effective traditional Chinese medicine composition for the treatment of influenza, which is of great significance, especially for influenza A and influenza B in children, which are 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, comprising the following medicinal ingredients in the following dosages by weight: 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 root, 4-8 parts of notopterygium wilfordii, 4-8 parts of isatis indigotica leaf, 4-8 parts of liushenqu, and 1-5 parts of liquorice.

[0008] Preferably, the medicinal ingredients and 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 root, 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:

[0010] This disease is often caused by wind-cold binding the exterior and internal heat accumulating. This disease presents numerous clinical manifestations, so the fundamental principle of clinical treatment should be to dispel cold and clear lung heat. Pueraria root is sweet, pungent, and cooling. It enters the lung, spleen, and stomach meridians; it dispels external pathogens, clears muscle heat, and allows the clear yang to ascend, moistening the tendons and veins. Ephedra is pungent, slightly bitter, and warming. It enters the lung and bladder meridians. It opens the closed skin and fur externally and restores the lung's normal function of clearing and descending. Their combined effects are mutually beneficial, so Pueraria root and ephedra are both monarch herbs. Gypsum is pungent and sweet, dispersing its pungent properties while its sweet properties regulate the middle, thus clearing lung heat and relieving cough and asthma. When combined with ephedra, one is cold and the other warm. Ephedra absorbs the coldness of gypsum, mitigating its warming and drying properties, while gypsum draws ephedra inward, slowing its diaphoretic effects. Apricot kernel is bitter and slightly warming, thus descending lung qi, which, when reduced, dissipates phlegm and stops cough. Paired with ephedra, ephedra promotes lung qi, while apricot kernel promotes and descends lung qi. The two herbs, one promoting and the other descending, complement each other, perfectly suiting the lung's function. Notopterygium wilfordii is pungent, bitter, and warm. It enters the lung and bladder meridians, boasting potent properties, promoting dispersing and relieving exterior qi, while also removing dampness and relieving pain. Bupleurum chinense is pungent, bitter, and slightly cold. It enters the liver, gallbladder, spleen, and lung meridians, dispersing wind-cold and effectively reducing fever. Isatis indigotica is bitter and cold. It enters the heart and stomach meridians. It enters the qi, clearing heat and detoxifying, and enters the blood, cooling blood and relieving sore throat. Liushenqu is sweet, pungent, and warm. It enters the spleen and stomach meridians. It promotes digestion, strengthens the spleen and stimulates appetite, and has a slight effect on relieving exterior symptoms and reducing fever, making it particularly suitable for exogenous symptoms combined with food stagnation. Licorice root is sweet and neutral. It enters the heart, lung, spleen, and stomach meridians. Its mild properties prevent ephedra from being too warm and dry, while also preventing gypsum from being too cold and damaging to the stomach. These herbs complement the deficiencies of the principal and auxiliary herbs, enhancing the formula's efficacy in dispelling exterior cold and clearing interior heat. This combination of herbs works together to dispel exterior cold and clear interior heat, eliminating pathogenic factors and promoting the smooth flow of Qi and blood, ultimately achieving the full therapeutic effect. The formula is meticulously formulated and appropriately combined, ensuring its proven efficacy in clinical practice.

[0011] The ratio of the formula of the present invention is also very important, and it has been proven to be effective within the dosage range of the formula 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 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: Add water to cover the API after filtering out the liquid in step S3, and repeat step S3 to filter out the second liquid;

[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 superficial cold and internal heat, or the influenza is influenza A or influenza B. Preferably, the influenza A is HIV-1 influenza A.

[0022] Preferably, the Chinese medicine composition is used in the preparation of a medicine for treating influenza or a cold 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. 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 symptoms is 91.52%. No complications, severe diseases or critical diseases occur.

[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, significantly reducing the viral load in the lung tissue of mice and significantly increasing the sIgA level in the upper respiratory tract lavage fluid; significantly reducing the levels of TNF-α, IL-2, IL-6, and IFN-γ in the serum of mice; significantly reducing the lung index and lung tissue lesion scores of each group of mice; and significantly reducing 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 directed to the syndrome of cold in the exterior and heat in the interior. This Chinese medicine composition has a mild medicinal property, preventing the excessive warming and drying of Ephedra, and preventing the coldness and cooling of Gypsum from damaging the stomach. The above medicines make up for the deficiency of monarch and minister, and strengthen the efficacy of this prescription in dispelling cold and clearing heat in the interior. This prescription is compatible with various medicines and plays the role of dispelling cold and clearing heat in the interior, so that the pathogenic factors are removed from the exterior and the qi and blood flow smoothly, and the efficacy is fully achieved. 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 Chinese medicine composition of the present invention is more reasonable in composition and has better efficacy, which proves that a synergistic effect has been generated 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 examples 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, comprising the following medicinal ingredients and dosages by weight: 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 wilfordii, 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 in water for 15 minutes, where the volume mass ratio of water to Chinese medicinal materials is 2 ml:1 g;

[0035] S3: first boil with high heat, then simmer for 25 minutes, and filter the liquid;

[0036] S4: Add water to cover the API after filtering out the liquid in step S3, and repeat step S3 to filter out the second liquid;

[0037] S5: The medicinal liquid filtered out in step S3 is mixed with the second medicinal liquid filtered out in step S4, and the mixture is concentrated to 50 mL to obtain the desired Chinese medicine composition.

[0038] Comparative Example 1

[0039] A traditional Chinese medicine composition comprises the following medicinal ingredients and dosages by weight: 3 parts of roasted ephedra, 20 parts of gypsum, 5 parts of bitter almond, 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 comprises the following medicinal ingredients and dosages by weight: 10 parts of kudzu root, 6 parts of bupleurum root, 5 parts of notopterygium wilfordii, 6 parts of isatis indigotica leaf, 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, comprising the following medicinal ingredients and dosages by weight: 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 wilfordii, 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 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 with high heat, then simmer for 20 minutes, and filter the liquid;

[0050] S4: Add water to cover the API after filtering out the liquid in step S3, and repeat step S3 to filter out the second liquid;

[0051] S5: The medicinal liquid filtered out in step S3 is mixed with the second medicinal liquid filtered out in step S4, and concentrated 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 conducted in accordance with national GLP standards to observe the acute toxic reactions and deaths in SD rats after oral administration of the drug solution of Example 1 three times within 24 hours, providing reference data for repeated dose toxicity testing.

[0054] The test selected 40 SPF-grade SD rats that passed the quarantine, half male and half female, and raised them in 475mm×350mm×200mm cages, with 5 rats in each cage. They were raised in accordance with the international (GB14925-2010) SPF-grade experimental animal environmental conditions, and the experimental animals were quarantined and acclimatized 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 male and half female, weighing 180.3-203.8g, were selected and randomly divided into 2 groups according to sex and 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 for more than 12 hours but not water, and then given pure water and Example 1 by oral gavage at 20 mL / kg. The animals were given 3 times on the day of administration (6 hours between administrations). Within 0 to 4 hours after each administration, the poisoning symptoms and characteristics, the occurrence of toxic reactions and the recovery time, as well as the mortality of the animals in each group were closely observed and recorded. The animals were then observed twice a day, once in the morning and once in the afternoon, for 14 consecutive days. The animals were weighed before administration on the day of administration and on the 4th, 7th, 10th, and 14th day after administration, and the weight changes and mortality of the animals were recorded.

[0055] Test results: Effects on general activity, poisoning symptoms, and mortality in experimental animals: Within 0 to 4 hours after each oral gavage administration, the rats in the blank control group and Example 1 group showed no significant abnormalities in their locomotor activity, mental state, or diet, and no related toxic reactions or deaths were observed. Continuous observation for 14 days after administration revealed no significant abnormalities in their locomotor activity, mental state, or diet, and no related toxic reactions or deaths were observed in the rats in the blank control group and Example 1 group.

[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 day after administration. There was no statistically significant difference in the body weight of the experimental animals in the Example 1 group compared with the blank control group during the same period, and the weight gain was within the normal growth range, indicating that oral administration of Example 1 to SD rats had no significant effect on the weight gain 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 doses). No 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] Three-month repeated administration toxicity test of Example 1 in rats by oral administration

[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 quarantine were selected, half male and half female, weighing 178.5-224.2 g, and measuring 475×350×200 mm. 3 The rats were raised in cages, with 5 rats per cage. The animals were raised in accordance with the national standard (GB14925-2010) SPF-level experimental animal environmental conditions, and the animals were quarantined and acclimatized for 7 days. They were randomly divided into 4 groups according to gender and weight, namely the blank control group, the low-dose group, the medium-dose group and the high-dose group of Example 1 (10, 20, 40g slices / kg), 20 animals in each group, and each group was gavage-administered at a volume of 15mL / 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.

[0064] Test results:

[0065] General Clinical Observations: During the dosing and recovery observation periods, the animals in each dose group of Example 1 were compared with the blank control group during the same period. No abnormalities related to drug toxicity were observed in the animals' appearance, behavioral activity, cavitary secretions and excretions, or general condition. During the trial, all animals were euthanized as planned, and no animals died abnormally during the trial.

[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 indicators 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 indices 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 the various dosage groups 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 the dose groups 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] Pharmacodynamic 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), which was 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: 13-15 g; (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), and interleukin 6 (IL-6) ELISA kits (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 N-terminal kinase (JNK) (Affinity), sodium pentobarbital (Shengxing Biotechnology Co., Ltd.); 4% paraformaldehyde (Wuhan Boster Bioengineering Co., Ltd.); HE staining solution kit (Wuhan Sevier Biotechnology Co., Ltd.).

[0102] 3 Grouping and modeling

[0103] Sixty 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 anesthesia with 1% pentobarbital, 25 μL of PBS was dripped into each nostril of each mouse in the blank group. 25 μL of the diluted virus solution was dripped into each nostril of each mouse in the remaining groups, with an infection dose of 20 mice at a median lethal dose (LD50).

[0104] 4. Dosage Method

[0105] Each group began dosing on the day of infection. The mice in Example 1 and Comparative Examples 1 and 2 were dosed at a clinically equivalent dose in mice (adult dose x 0.0026 x 50 = 8.19 g crude drug / kg) based on body surface area. The oseltamivir phosphate group was dosed at 27.5 mg / kg, converted to an adult clinical equivalent dose. The blank and model groups were administered distilled water, and mice in each group were gavaged once daily for three consecutive days at a dose of 0.2 mL / 10 g.

[0106] 5 Observation indicators and detection methods

[0107] ELISA was used to detect the levels 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 with 0.3% sodium pentobarbital (0.2 ml / 10 g) intraperitoneally. Blood was collected from the mouse eyeballs and placed in EP tubes. After standing at 4°C for 30 minutes, the blood was centrifuged at 4°C, 3000 r / min (centrifugal radius 14 cm) for 15 minutes. The serum was collected and stored at -80°C until testing. The mouse trachea was exposed by thoracotomy. 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 three times. The upper respiratory tract lavage fluid was collected and then centrifuged at 4°C, 3000 r / min (centrifugal radius 14 cm) for 15 minutes. The supernatant was collected and stored at -80°C until testing. The levels of sIgA in upper respiratory tract lavage fluid and serum IFN-γ, TNF-α, IL-2, and IL-6 were detected strictly according to the ELISA kit instructions.

[0108] RT-qPCR was used to assess lung viral load. 15-20 mg of right lung tissue from mice was ground and centrifuged at 4°C, 3000 rpm (centrifuge radius, 14 cm), for 15 minutes. The supernatant was collected as a lung tissue homogenate. Total RNA was extracted using TRIzol and the protocol was followed according to the instructions of the influenza A virus nucleic acid detection kit. PCR amplification was performed using target gene primers (parameters: 48°C for 5 minutes, 94°C for 2 minutes, 94°C for 10 seconds, 55°C for 35 seconds, 40 cycles). PR8 mRNA levels were measured, and copy number was calculated using a scanning method based on a standard curve.

[0109] HE staining was used to observe lung tissue pathological changes. Left lobe lung tissue was fixed in 4% paraformaldehyde, dehydrated with graded alcohol, transparentized with xylene, embedded in paraffin, sectioned, and stained with HE. The slides were then examined under a light microscope. Lung tissue lesions were scored based on the degree of lung parenchymal lesions, interstitial lung inflammation, and peribronchial inflammation. A score of 0 indicated healthy lungs without lesions, while scores of 1, 2, 3, and 4 indicated mild interstitial pneumonia (<25% of the lung), moderate damage (25%-50% of the lung), severe damage (51%-75% of the lung), and very severe damage (>75% of the lung), respectively, were assigned.

[0110] The relative protein expression of TNFR1, NF-κB, and JNK1 in lung tissue was determined by Western blotting, which included 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. Normality and homogeneity of variance tests were performed when comparing multiple groups. If normality was met (P>0.05), statistical analysis was performed using one-way analysis of variance (ANOVA). Depending on the homogeneity of variance, LSD+Dunnet (for homogeneous variances) or Tamhane's T2 (for unequal variances) were used for comparative analysis. If normality was not met (P<0.05), the Kruskal-Wallis test was used, and pairwise comparisons were performed using the Mann-Whitney test. Statistical results were tested with an α=0.05 threshold, with P<0.05 indicating statistical significance. 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 decreased (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 both were statistically significant (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 showed that under the experimental conditions of this invention, 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 mice of each group ( 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 the comparative example 1 group, the comparative example 2 group, the example 1 group, and the positive control group were significantly decreased (P < 0.05 or P < 0.01); compared with comparative example 1 and comparative example 2, there were statistical differences (P < 0.05 or P < 0.01); compared with the positive control group, there was no significant difference in example 1, no statistical difference (P > 0.05).

[0122] Experimental Conclusion

[0123] The experimental results showed that under the experimental conditions of the present invention, the serum TNF-α, IL-2, IL-6 and IFN-γ levels 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 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 the 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 the mice in the comparative example 1 group, the comparative example 2 group, the example 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 example 1 (P < 0.05 or P < 0.01); compared with the positive control group, there was no significant difference in example 1, no statistical difference (P > 0.05).

[0129] Experimental Conclusion

[0130] The experimental results showed that under the experimental conditions, the lung index and lung tissue lesion score of mice in Example 1, Comparative Examples 1 and 2 groups and the positive control were significantly reduced (P < 0.05 or P < 0.01), indicating that Example 1 and Comparative Examples 1 and 2 had 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 had 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 levels of TNFR, NF-κB, and JNK1 proteins in the lung tissues of the model group mice were significantly increased (P < 0.01). Compared with the model group, the relative expression levels of TNFR, NF-κB, and JNK1 proteins in the lung tissues of the comparative example 1 group, the comparative example 2 group, the example 1 group, and the positive control group were significantly decreased (P < 0.05 or P < 0.01); there was a statistical difference between Example 1 and comparative examples 1 and 2 (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 showed that under the experimental conditions, the relative expression levels of TNFR, NF-κB, and JNK1 proteins in the lung tissue 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 had 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 Examples 1 and 2, Example 1 had a better effect.

[0137] Clinical observation data

[0138] Experimental 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 efficacy 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 enrolled 120 children with influenza and randomly divided them into a control group and an observation group, 60 cases each. The observation group consisted of 32 males and 28 females, aged 2 to 13 years (mean, 5.67±1.63) years; the duration of illness ranged from 1 to 5 days (mean, 3.02±1.03) days. The control group consisted of 33 males and 27 females, aged 2 to 14 years (mean, 5.58±1.90) years; the duration of illness ranged from 1 to 6 days (mean, 3.15±1.05) days. Comparison of the general data between the two groups showed no statistically significant differences (P>0.05), indicating comparability.

[0141] Diagnostic criteria

[0142] (1) Western medicine diagnostic standards

[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)." Diagnosis of influenza in children is primarily based on epidemiological history, clinical manifestations, and etiological testing.

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

[0145] (2) Clinically diagnosed cases: patients with the above-mentioned clinical manifestations of influenza, 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 patients 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 double serum samples from the acute phase and the recovery phase 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 breaths / min for children over 5 years old; >40 breaths / min for 1-5 years old; >50 breaths / min for 2-12 months old; >60 breaths / min for newborns up to 2 months old; ② Altered mental status: slow reaction, drowsiness, agitation, convulsions, etc.; ③ Severe vomiting, diarrhea, and signs of dehydration; ④ Oliguria: urine output <0.8 mL / (kg·h) for children, or <200 mL / m2 for infants and young children. 2 , preschool children <300 mL / m 2 , school-age children <400 mL / m 2 ⑤ Children over 14 years old have a blood pressure of <17 mL / 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] (2) TCM syndrome differentiation criteria (exterior cold and interior heat syndrome)

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

[0151] Main symptoms: ① fever, ② aversion to cold (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 fur, rapid pulse.

[0154] The main symptoms of the child must all be met, and two or more secondary symptoms must be met. 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 superficial cold and internal heat syndrome;

[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) duration of illness ≤ 48 hours (the duration of illness is defined as the time from onset to the current visit, and the onset time refers 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, appears);

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

[0162] (2) 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 pharyngoconjunctival fever, 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 medicine for treating 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 those with 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 do not subside 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 subject suffers from other diseases that affect the judgment of efficacy and safety;

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

[0178] 5) If the subject's condition worsens or complications develop, such as pneumonia, after medication, the medication should be stopped and effective treatment measures should be taken. The subject should complete all laboratory tests, withdraw from the trial, and be treated as an ineffective case;

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

[0180] 7) Those whose viral nucleic acid test results were 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 received medication or 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 (a Chinese medicine composition of the present invention) is composed of 9 herbs: Pueraria root, roasted ephedra, gypsum, bitter almond, bupleurum, notopterygium root, isatis indigotica leaf, 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 weighing less than or equal to 15 kg, take 30 mg 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 weighing less than or equal to 23 kg and greater than or equal to 40 kg, take 60 mg each time, 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°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 if the fever symptoms do not subside for more than 24 hours, the subject is advised to withdraw from the trial.

[0202] (2) Except for the experimental drugs and 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 used by the subjects should be recorded in the research medical records, including the generic name, time of use, dosage, and reason for use of the concomitant medications, and a determination should be made as to whether they affect the efficacy of the trial medications.

[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] Screening indicators

[0212] C-reactive protein (CRP) and chest CT / X-ray were performed at the investigator's discretion.

[0213] Diagnostic indicators

[0214] Throat swab test

[0215] 1) First, rapid antigen detection method (colloidal gold method) is selected for screening;

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

[0217] Effectiveness evaluation

[0218] Observation indicators

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

[0220] (2) General clinical information: personal history, past medical history, physical examination, concurrent diseases and medications, trial start date, etc.

[0221] (3) Efficacy indicators:

[0222] ① Complete fever reduction time (hours). After the first treatment, 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 symptoms (cure and significant improvement rate) was scored at baseline and 5 days after treatment, and evaluated 5 days after 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 was performed on throat swabs at baseline and 5 days after treatment, and the evaluation was performed 5 days after 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, 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] Clinically cured: symptoms have basically disappeared, and the TCM syndrome score has decreased by ≥95%;

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

[0234] Effective: Symptoms improved, with a decrease rate of TCM syndrome score ≥30% and <70%;

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

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

[0237] Quantitative Grading Standards for Traditional Chinese Medicine Syndrome

[0238] This study was developed 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 Implementation).

[0239] Table 13 Quantitative standard for grading symptoms of TCM syndrome of cold in the exterior and heat in the interior

[0240]

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

[0242] Statistical analysis

[0243] 1. Definition of the dataset

[0244] The Full Analysis Set (FAS) included all subjects who were randomized, received at least one dose, and had at least one visit. The intention-to-treat (ITT) analysis was performed using the FAS. For missing values ​​in the primary efficacy outcome measures, the most recent observation carried forward to the final trial outcome (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): The actual data of patients who have received at least one treatment and have safety indicators recorded. Data of patients who have withdrawn 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 Missing Values

[0249] For missing values ​​of efficacy indicators, the most recent observation data will be carried forward to the final trial results (LOCF method). Missing values ​​of safety indicators will not be carried forward.

[0250] 3. General Principles of Statistics

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

[0252] For time series data, median survival time was described and Kaplan-Meier curves were plotted. The log-rank test was used to compare the difference in median survival time between the two groups. Cox regression analysis was used to consider the influence of center or other factors.

[0253] Qualitative data should describe the number and percentage of cases in each category. For enumeration data, the chi-square test and Fisher's exact test were used for comparisons between treatment groups. For ranked data, the Wilcoxon rank-sum test was used for comparisons between treatment groups. If the influence of center or other factors was considered, the CMH chi-square test or logistic regression was used.

[0254] Statistical analysis was performed using SAS v9.4 software. Two-sided tests were used, with α = 0.05. All confidence intervals were calculated at a 95% confidence level.

[0255] 4. Multiplicity Adjustment

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

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

[0258] Statistical results

[0259] (1) Study enrollment

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

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

[0262] (2) Baseline comparability analysis: FAS and PPS analysis

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

[0264] There were no statistically significant differences in the efficacy-related indicators, including the baseline total score of TCM syndromes and individual symptom scores, between the two groups. The conclusions of FAS and PPS were consistent.

[0265] (3) 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 it takes for flu symptoms to resolve in the two groups showed a median of 48 hours in the test group and 80 hours in the control group. Log-Rank comparisons between the two groups showed statistically significant differences. The conclusions of FAS and PPS were consistent.

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

[0272]

[0273] 2. Time to completely reduce fever (hours)

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

[0275] Table 15 Comparison of complete fever reduction time

[0276]

[0277] 3. Therapeutic effect of TCM syndromes

[0278] Excluding the interaction between center and group, the FAS analysis of the covariance of the TCM syndrome score reduction at the endpoint, using the baseline TCM syndrome score as a covariate, showed no statistically significant differences between the two groups in the TCM syndrome score reduction at the endpoint. Specifically, the Lsmean of the TCM syndrome score reduction at the endpoint was 9.3119 points in the experimental group and 9.0953 points in the control group (P = 0.3497).

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

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

[0281] Using the FAS dataset, the inter-group comparison of the disappearance time of fever, headache and body pain, sore throat, runny nose, cough, fatigue, and thirst showed that the experimental group had a shorter disappearance time than the control group, with a statistically significant difference (P < 0.05). The inter-group comparison of the disappearance rate of fever and cough, using the FAS dataset, showed that the disappearance rate of the above individual symptoms was higher in the experimental group than in the control group, with a statistically significant difference (P < 0.05).

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

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

[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 embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any technician familiar with the technical field, within the technical scope disclosed by the present invention, who makes equivalent replacements or changes based on the technical solutions and concepts of the present invention, should be covered by the scope of protection of the claims of the present invention.

Claims

1. A Chinese medicine composition for treating influenza, characterized in that: The medicinal ingredients 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 wilfordii, 4-8 parts of isatis indigotica, 4-8 parts of liushenqu, and 1-5 parts of liquorice. The influenza is a syndrome of cold on the surface and heat on the inside, or the influenza is influenza A or influenza B.

2. The Chinese medicine composition for treating influenza according to claim 1, characterized in that: The medicinal ingredients and dosage by weight are as follows: 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 wilfordii, 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 ratio of components and contents; S2: The mixed Chinese herbal raw materials are placed in a casserole and soaked in water for 10-30min, wherein the volume mass ratio of water to Chinese herbal 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 further comprises: 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 traditional 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 superficial cold and internal heat, 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 to 14 years old.

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