Application of traditional Chinese medicine composition in preparation of medicine for treating influenza and influenza caused by viruses

Abstract

The invention discloses application of a traditional Chinese medicine composition in preparation of a medicine for treating influenza and influenza caused by viruses, the traditional Chinese medicine composition comprises the following traditional Chinese medicine raw materials in parts by weight: 5-35 parts of bupleurum chinense, 9-50 parts of scutellaria baicalensis, 9-50 parts of artemisia apiacea, 5-35 parts of red paeony root, 5-35 parts of moutan bark and 4-26 parts of perilla leaf. The traditional Chinese medicine is used for treating influenza caused by viruses and fever, hyperpyrexia, headache, body pain, nasal obstruction, yellow and turbid nasal discharge, swollen sore throat, cough, sticky or yellow phlegm and fatigue and weakness of the whole body caused by influenza, so that the cold and the influenza are effectively treated.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine technology, specifically relating to the application of a traditional Chinese medicine composition in the preparation of a drug for treating colds and influenza caused by viruses. Background Technology

[0002] The common cold is caused by the invasion of the human body by the six pathogenic factors and seasonal viruses.

[0003] The term "common cold" originates from the Northern Song Dynasty text *Ren Zhai Zhi Zhi Fang Qing Feng*, which, in its section on "Treatise on Common Cold Prescriptions," quotes *He Ji Ju Fang Shen Su Yin*, stating that it treats common cold with wind-heat, fever, headache, heavy cough, and thick, sticky nasal discharge. The *Su Wen Feng Lun* states, "Wind injures people, sometimes causing chills and fever." Zhang Zhongjing's *Shang Han Lun Bian Taiyang Bing Mai Zheng Bing Zhi* from the Han Dynasty laid the foundation for the diagnosis and treatment of common cold when discussing Taiyang disease. *Zhu Bing Yuan Hou Lun. Shi Qi Bing Zhu Hou* states, "Due to seasonal imbalances and irregular temperatures, people are susceptible to illness caused by adverse weather conditions, and these illnesses are easily transmitted." The common cold encompasses both the common cold (wind-cold) and influenza (seasonal influenza). Traditional Chinese medicine believes the cause of the common cold is the invasion of the body by the six pathogenic factors (wind, cold, dampness, heat, and dampness). Wind is often the first of the six pathogenic factors: wind combined with cold is called wind-cold syndrome, wind combined with heat is called wind-heat syndrome, and summer heat combined with dampness is called wind-summer-dampness syndrome. Seasonal influenza is epidemic and contagious. Significant fever and high fever symptoms indicate the presence of wind-heat pathogens, which cause heat to stagnate in the muscles and skin, disrupting the balance of the defensive qi and impairing the lung's function of clearing and purifying. Pathogenic factors are a major contributor to the common cold.

[0004] Traditional Chinese Medicine (TCM) Symptom Analysis: Wind-heat pathogens invade the body surface, causing a struggle between the pathogenic factors and the body's resistance, resulting in fever. Disruption of the opening and closing of the pores and imbalance of the body's defensive and nutritive qi lead to mild aversion to wind and cold, general malaise, and reduced sweating. Wind-heat pathogens deplete body fluids, causing thirst. Wind-heat pathogens ascend and easily attack the yang organs, causing headache and sore throat. Although initially superficial, the wind-heat pathogens rapidly transform. As they penetrate deeper, symptoms such as dry throat appear, indicating a progression to a semi-exterior, semi-interior condition. Wind-heat damages the lungs (fire overpowering metal), impairing the lung's function of dispersing and descending qi, resulting in severe cough and thirst. The wind-heat pathogens invading the lungs affect their dispersing and descending functions, causing cough. The heat depletes body fluids, forming phlegm, resulting in thick, sticky phlegm. The wind-heat pathogens affect the nasal passages, causing congestion and edema of the nasal mucosa, resulting in nasal congestion and thick, turbid nasal discharge. The wind-heat pathogens ascend and damage the throat, causing sore throat.

[0005] Influenza (flu) is an acute respiratory infectious disease caused by the influenza virus. It has a seasonal pattern of occurrence, but can also circulate year-round. The general population is susceptible, but pregnant women, infants, the elderly, and those with chronic underlying diseases are more likely to develop severe or critical cases, resulting in a higher mortality rate.

[0006] Influenza symptoms: It begins with fever, headache, muscle pain and general malaise. The body temperature reaches 39-40 degrees Celsius. There may be chills, shivering, and often accompanied by systemic symptoms such as muscle and joint pain, fatigue, and loss of appetite. Sore throat, dry cough, nasal congestion, runny nose, facial flushing, and conjunctival congestion may also occur.

[0007] The pathological changes are characterized by insufficient vital energy, weakened defensive function, and lax regulation of the lungs and wei qi. This allows pathogenic factors, including the six external evils and viral infections, to invade through the mouth, nose, skin, and hair, with the lungs and wei qi bearing the brunt. Symptoms of disharmony between the defensive qi and the lungs' inability to disperse and descend qi quickly appear. Alternatively, improper lifestyle, imbalances in temperature, and overwork can lead to weakened muscle defenses, making the body susceptible to external pathogens.

[0008] Traditional Chinese medicine emphasizes syndrome differentiation and treatment, adhering to the principle that "prescriptions are derived from treatment methods, and treatment methods are established according to syndromes." Different combinations of prescriptions, based on their own treatment principles, have different principal, assistant, adjuvant, and guide ingredients, and target different pathologies and mechanisms, resulting in significant differences.

[0009] Based on clinical practice and analysis of the symptoms of common cold (wind-heat attacking the exterior) and influenza, the clinical value of this invention is as follows: Function: Clears heat and detoxifies, expels pathogens from the exterior. Symptoms include: fever, high fever, headache, body aches, nasal congestion with thick yellow nasal discharge, sore throat, cough, sticky or yellow phlegm, and general fatigue.

[0010] Based on clinical practice, current treatments for upper respiratory tract infections target symptoms such as fever, cough, nasal congestion, dry throat, and sore throat. Western medicine treatments, including antipyretics, antivirals, and antibiotics, are often ineffective because wind-heat pathogens easily damage body fluids. A slight sweating after medication is recommended, as excessive sweating can further deplete body fluids and worsen the condition. Furthermore, the use of Western antipyretics, analgesics, and steroid antipyretics also leads to excessive sweating and depletion of body fluids. The antiviral drug prepared in this invention addresses this issue by promoting slight sweating, thus overcoming the shortcomings of Western antipyretics. Western antiviral treatments are slow to take effect and lack timely intervention. Moreover, the overuse of antibiotics, frequent changes in antibiotics, and the use of broad-spectrum antibiotics often suppress and kill normal flora, leading to fungal infections, increasing drug-resistant bacteria and hospital-acquired infections, and causing immunodeficiency in patients. Therefore, rational drug use is crucial for alleviating patients' clinical symptoms.

[0011] The composition of this invention combines the inheritance of traditional Chinese medicine theory with innovation supported by modern scientific and technological evidence. It is also supported by modern pharmacodynamic experimental data and verified by clinical applications. It cannot be easily obtained through conventional additions or subtractions of medicinal ingredients. Summary of the Invention

[0012] The technical problem to be solved by the present invention is to provide an application of a traditional Chinese medicine composition in the preparation of a drug for treating colds and influenza caused by viruses.

[0013] Traditional Chinese medicine (TCM) has unique characteristics and a rich clinical foundation in the prevention and treatment of colds and influenza. Based on TCM clinical analysis of the clinical characteristics of colds and influenza, combined with traditional TCM theory and the features of this invention, the clinical application of colds (wind-heat syndrome) and influenza is more precise and clinically valuable.

[0014] The application of this invention is based on traditional Chinese medicine theory and follows traditional Chinese medicine principles and practices. The application of this invention is analyzed as follows:

[0015] 1. Common cold, influenza. Clinically speaking, the main symptom is fever. The main treatment combination is Artemisia annua, Scutellaria baicalensis, and Bupleurum chinense, which have heat-clearing and antiviral properties.

[0016] 2. From the perspective of differentiating syndromes based on the Wei Qi Ying Xue (defense, qi, nutritive, and blood) principles for the common cold and influenza: Heat pathogens and evil qi permeate the Qi, Ying, and blood levels. Clearing heat from the Qi, Ying, and blood levels simultaneously is crucial. When heat pathogens and evil qi ascend, they first attack the lungs, spreading from the interior to the exterior. Use peony bark and red peony root to clear heat, cool the blood, detoxify, and clear heat from the Ying and blood levels. Interrupt the development of the febrile disease, prevent its spread, and avoid worsening of the condition.

[0017] 3. Treating Pathogenic Factors in the Lesser Yang Channel: The *Suwen* states, "Ascending, descending, entering, and exiting—all things exist within this channel." The pivot, like the axis of a doorway, functions through the ascending, descending, entering, and exiting of Qi. Regulating Qi is key to the pivot, especially the Lesser Yang pivot. The Lesser Yang relies on the pivot for its function, indicating its crucial role in regulating the flow of Qi between the interior and exterior, and ensuring smooth Qi circulation. When pathogenic factors invade the Lesser Yang, its Qi circulation is impaired, leading to stagnation, heat, and fire. This invention addresses this by starting with the Lesser Yang: Artemisia annua, Scutellaria baicalensis, and Bupleurum chinense, which promote the flow of Qi, clear heat, moisten, and eliminate turbidity, thus clearing and facilitating the Lesser Yang.

[0018] 4. Viral infections fall under the categories of febrile diseases and epidemic diseases in Traditional Chinese Medicine (TCM). Based on the diagnostic system of Wei Qi Ying Xue and the San Jiao (Triple Burner), febrile diseases are a unique TCM infectious disease. Direct methods to kill the virus include: dispelling, resolving, eliminating, and dispersing. Indirect methods include raising Yang and dispersing heat, using sweet and warm herbs to clear heat, tonifying Qi and relieving exterior symptoms, and replenishing Qi and supporting detoxification. A holistic approach is adopted, focusing on eliminating pathogens and protecting Qi. Treatment is based on syndrome differentiation: seeking the cause to shorten the course of the disease; at different stages, medications for febrile diseases are used to inhibit and eliminate viruses, employing cooling and relieving exterior symptoms, clearing heat and detoxifying, clearing heat and cooling blood, and clearing heat and opening the orifices, all with a holistic protective awareness.

[0019] This invention applies the principles of "Wenbing Theory" and "Wei-Qi-Ying-Xue Differentiation," targeting the core pathogenesis of influenza: "wind-heat epidemic toxins attacking the exterior and accumulating internally in the lungs and stomach." It aims to cause the pathogenic toxins to penetrate deeper, stagnating and transforming into toxicity. Following the treatment principle of Ye Tianshi's "sweating is effective when the disease is at the Wei level, but clearing the Qi is necessary when it reaches the Qi level" (from *Wenre Lun*), it incorporates Wu Jutong's medical philosophy of "treating the upper burner like a feather, requiring gentleness to lift" (from *Wenbing Tiaobian*). The combined use of various herbs works synergistically to clear heat, detoxify, and promote the flow of Qi to the exterior. Furthermore, it heavily utilizes bitter and cold Scutellaria baicalensis and Artemisia annua for dispersion and clearing. These two herbs are the principal herbs.

[0020] In this formula, Scutellaria baicalensis and Artemisia annua are the principal herbs. Scutellaria baicalensis, bitter and cold in nature, enters the lung, gallbladder, and stomach meridians. The *Compendium of Materia Medica* states that it "treats phlegm-heat, benefits the chest and diaphragm, and treats pulmonary atrophy with cough and bloody pus." Zhang Yuansu's *Medical Origins* states that "Scutellaria baicalensis clears lung heat and treats damp-heat in the upper burner." Modern research shows that its active ingredient, baicalin, can significantly inhibit the activity of influenza virus neuraminidase (Wang Jinhua et al., 2015, *Chinese Journal of Traditional Chinese Medicine*). Artemisia annua, bitter and pungent in nature, is cold in nature. The *Essentials of Materia Medica* states that it "treats malaria with phlegm-cough, deficiency-heat, and night sweats." The Artemisia annua and turtle shell decoction in *Treatise on Febrile Diseases* uses Artemisia annua to expel heat. Its antiviral mechanism aligns with Wu Youke's treatment principle in *Treatise on Epidemic Febrile Diseases*, which emphasizes "expelling pathogens as the primary principle."

[0021] Bupleurum, Paeonia lactiflora, and Paeonia suffruticosa are the assistant herbs. Bupleurum is listed as a superior herb in the *Shennong Bencao Jing* (Shennong's Classic of Materia Medica), "treating stagnation of qi in the heart, abdomen, intestines, and stomach, food accumulation, and cold and heat evil qi." Its combination with Scutellaria baicalensis follows the principle of "harmonizing the lesser yang" in the *Shanghan Lun* (Treatise on Cold Damage), as Ke Qin stated: "Bupleurum soothes the wood element, Scutellaria baicalensis clears the metal element; this is a key remedy for the lesser yang" (*Shanghan Laisu Ji*). Paeonia lactiflora, according to *Yaopin Huayi* (Medical Properties Explained), "drains liver fire, eliminates blood stasis, and disperses stagnant blood." Combined with Bupleurum, it subtly aligns with Wang Qingren's Xuefu Zhuyu Tang (Blood-Stasis-Expelling Decoction) method of "regulating both qi and blood" (*Yilin Gaicuo*). Modern research confirms that its paeoniflorin can downregulate influenza virus-induced TNF-α overexpression (Li Chun et al., 2018, *Chinese Journal of Experimental Traditional Medical Formulae*). Paeonia suffruticosa, according to *Benjing Shuzheng* (Commentary on the Classic of Materia Medica), "treats cold and heat stroke, convulsions and epilepsy." Zhang Zhongjing's Shenqi Wan (Kidney Qi Pill) uses Paeonia suffruticosa to clear deficiency heat. This formula utilizes the property of "entering the blood and purging latent fire" (Zhang Xichun's "Medical Records of Integrating Chinese and Western Medicine") to prevent heat toxins from invading the blood.

[0022] Perilla leaf is used as an adjuvant herb. The *Compendium of Materia Medica* states that it "disperses cold qi, clears lung qi, and soothes the middle qi." This formula utilizes its three effects: ① As an adjuvant: preventing the bitter and cold herbs Scutellaria baicalensis and Artemisia annua from harming the stomach, reflecting the principle of "no harm if there is a cause" in the *Inner Canon of Medicine*; ② Guiding the medicine: its pungent and aromatic nature allows it to reach the exterior, guiding the medicine to the lung and wei levels, as Xu Lingtai stated: "All light and weak medicines can enter the lungs" (*On the Origin and Development of Medicine*); ③ Harmonizing the stomach: addressing the common "lung and stomach disease" symptoms of influenza, practicing Ye Tianshi's academic thought of "treating the middle to treat the upper disease."

[0023] The innovation of this invention, based on inheritance, lies in:

[0024] 1. The formula employs a comprehensive approach that does not simply relieve exterior symptoms, nor simply clear interior heat. It clears heat and detoxifies through dispersing and harmonizing, while avoiding the use of warming agents that could worsen the condition.

[0025] 2. Emphasize the use of heat-clearing and detoxifying drugs to eliminate the cause of the disease, thus avoiding the situation where the medicine is too weak to treat the severe illness. Also, avoid the indiscriminate use of bitter and cold drugs, which may trap the pathogenic factors or cause the body to cool down but the function to be difficult to recover, resulting in a prolonged and difficult-to-cure condition. Because the pathogenic factors are trapped inside and become stagnant and turn into toxins, the treatment adopts the method of pungent and dispersing, clearing and promoting circulation, cooling without stagnation, which reflects the principle of "the upper burner being as light as a feather".

[0026] 3. Because wind-heat pathogens easily damage body fluids, the medication should only induce a slight sweat. Excessive sweating can further damage body fluids, leading to complications and worsening of the condition. Furthermore, the use of Western antipyretics and analgesics, as well as steroid antipyretics, can cause excessive sweating and damage body fluids. This formula induces a slight sweat, thus mitigating the shortcomings of Western medications.

[0027] 4. Wind-heat pathogens and influenza pathogens spread rapidly. Since the pathogen resides in the lungs and defensive qi, efforts should be made to cure it and prevent its further spread. Sufficient medication should be used to eliminate the pathogen. Based on antipyretic and detoxifying principles, peony bark and red peony root should be used to interrupt the spread of the pathogen.

[0028] Modern scientific research has also demonstrated the scientific validity of this invention through its effective material basis.

[0029] The Western medical pathogenesis of the common cold and influenza is as follows: weakened immune function and physiological structure are the internal conditions, while the virulence of the pathogen is the external condition leading to the onset of the disease. After the pathogen invades the respiratory tract, it multiplies locally, directly damaging the respiratory organs or releasing toxic substances. The body, due to the invasion of the pathogen, damages and induces immune responses and cytokines, causing pathological processes.

[0030] Modern scientific research has provided evidence for the antiviral effects of the medicinal ingredients used in this invention.

[0031] The main active components of Bupleurum chinense are saikosaponins and polysaccharides. Studies have shown that saikosaponins A / D can inhibit viral replication by blocking the nuclear export of influenza virus (H1N1) nucleoprotein (NP) (Li et al., 2017). Furthermore, polysaccharides can enhance the clearance capacity of macrophages against respiratory syncytial virus (RSV) by activating the TLR4 / NF-κB signaling pathway (Zhu et al., 2016). Clinical studies have shown that Bupleurum chinense injection combined with conventional treatment can significantly shorten the duration of fever in children with influenza (Zhang et al., 2019), suggesting its potential application value in antiviral therapy.

[0032] The antiviral activity of Scutellaria baicalensis is mainly attributed to baicalin and baicalein. Baicalin can inhibit the activity of the 3CL protease of SARS-CoV-2, thereby blocking viral replication (Liu et al., 2020). Baicalein has an inhibitory effect on the NS5 protein of dengue virus (DENV) (Chen et al., 2018). In clinical applications, baicalin injection can effectively relieve the pain symptoms of patients with herpes zoster (Wang et al., 2021), indicating its potential in the fight against herpesviruses.

[0033] Artemisinin and its derivatives (such as dihydroartemisinin) are the main active components of Artemisia annua. Studies have found that artemisinin can inhibit the replication of hepatitis B virus (HBV) by inducing the generation of reactive oxygen species (ROS) (Li et al., 2019), while dihydroartemisinin can block the UL12 nuclease activity of herpesvirus-1 (HSV-1) (Zhou et al., 2020). Clinical trials have shown that artesunate combined with interferon can improve the HBeAg seroconversion rate in the treatment of chronic hepatitis B (Yang et al., 2018), supporting its application in antiviral therapy.

[0034] The active components of Paeonia lactiflora include paeoniflorin and gallic acid. Studies have shown that paeoniflorin can alleviate influenza virus-induced lung inflammation by inhibiting the TLR7 / MyD88 signaling pathway (Ding et al., 2018). Furthermore, Paeonia lactiflora extract has a protective effect against Coxsackievirus B3 (CVB3)-induced myocarditis (Liu et al., 2017). In the treatment of COVID-19, compound formulas containing Paeonia lactiflora (such as Qingwen Baidu Decoction) can reduce the levels of inflammatory factors in patients (Xiong et al., 2020), suggesting its antiviral and immunomodulatory effects.

[0035] Paeonol and paeoniflorin are the main active components of peony bark. Paeonol can inhibit the activation of the NLRP3 inflammasome, thereby alleviating lung damage caused by influenza virus (Zhu et al., 2019). In vitro experiments have also found that peony bark extract has an inhibitory effect on HIV-1 integrase (Kong et al., 2016). Clinical studies have shown that paeonol ointment can be used as an adjunct treatment for postherpetic neuralgia (Li et al., 2020), indicating its potential value in anti-herpes virus activity.

[0036] The main active components of perilla, perillaldehyde and rosmarinic acid, exhibit significant antiviral effects. Perillaldehyde can inhibit influenza A virus (H5N1) infection by disrupting the viral envelope (Wu et al., 2018), while rosmarinic acid can block the NS2B-NS3 protease activity of Zika virus (ZIKV) (Dong et al., 2021). Clinical trials have confirmed that perilla leaf extract can shorten the duration of diarrhea in children with rotavirus enteritis (Chen et al., 2019), supporting its application in the treatment of gastrointestinal viral infections.

[0037] The aforementioned six traditional Chinese medicines and their active ingredients exert antiviral effects by directly inhibiting viral replication (e.g., baicalin targeting 3CL protease) or regulating host immune responses (e.g., Bupleurum polysaccharide activating the TLR4 pathway). Current research mainly focuses on pathogens such as influenza virus, herpesvirus, hepatitis virus, and SARS-CoV-2, and some components have entered the clinical validation stage.

[0038] The compositions of this invention have achieved good clinical and experimental results in clinical applications and modern pharmacodynamic studies.

[0039] To address the aforementioned technical problems, this invention provides an application of a traditional Chinese medicine composition in the preparation of a drug for treating colds and influenza caused by viruses. The weight ratio of the various components is as follows: Bupleurum chinense 5-35 parts, Scutellaria baicalensis 9-50 parts, Artemisia annua 9-50 parts, Paeonia lactiflora 5-35 parts, Paeonia suffruticosa 5-35 parts, and Perilla frutescens 4-26 parts.

[0040] In the aforementioned traditional Chinese medicine composition, the preferred weight ratio of each component is as follows: Bupleurum chinense 8-28 parts, Scutellaria baicalensis 12-40 parts, Artemisia annua 12-40 parts, Paeonia lactiflora 8-28 parts, Paeonia suffruticosa 8-28 parts, and Perilla frutescens 8-22 parts.

[0041] In the aforementioned traditional Chinese medicine composition, the preferred weight ratio of each component is as follows: Bupleurum chinense 10-25 parts, Scutellaria baicalensis 20-35 parts, Artemisia annua 20-35 parts, Paeonia lactiflora 10-25 parts, Paeonia suffruticosa 10-25 parts, and Perilla frutescens 12-20 parts.

[0042] The aforementioned traditional Chinese medicine composition can be prepared into any commonly used formulation.

[0043] The traditional Chinese medicine composition can be used to prepare preparations including tablets, capsules, granules, pills, mixtures, syrups, decoctions, powders, suppositories, gels, sprays, or injections.

[0044] The Chinese medicines described in this invention can be replaced by Chinese medicines with the same or similar effects, and these medicinal materials can all be processed in accordance with the "National Standards for Processing Chinese Medicines" or the "Dictionary of Chinese Materia Medica". For example, Scutellaria baicalensis can be replaced by Scutellaria baicalensis processed with wine, Bupleurum chinense can be replaced by Bupleurum chinense, and Paeonia lactiflora can be replaced by Paeonia veitchii. The efficacy of the replaced medicinal materials can be better than that of the original medicinal materials, but all of them are within the scope of this invention.

[0045] To address the aforementioned technical problems, this invention provides an application of a traditional Chinese medicine composition in the preparation of a drug for treating colds and influenza caused by viruses. The preparation method includes the following steps:

[0046] ① Weigh out the following raw materials according to the proportions: 5-35 parts of Bupleurum chinense, 9-50 parts of Scutellaria baicalensis, 9-50 parts of Artemisia annua, 5-35 parts of Paeonia lactiflora, 5-35 parts of Paeonia suffruticosa, and 4-26 parts of Perilla frutescens.

[0047] ② Take Bupleurum chinense, Scutellaria baicalensis, Artemisia annua, Paeonia lactiflora, Paeonia suffruticosa, and Perilla frutescens leaves, add 6 to 14 times the amount of raw herbs to water, heat and reflux for 0.5 to 5 hours, heat and reflux for 1 to 3 times, filter, combine the extracts, and concentrate under reduced pressure at 40 to 90℃ to a density of 1.01 to 1.20 for later use;

[0048] ③Dry the concentrated liquid from step ② at 50-90℃, pulverize the dried paste, add excipients, and prepare the formulation.

[0049] To address the aforementioned technical problems, the present invention provides a method for preparing a traditional Chinese medicine composition, the method comprising the following steps:

[0050] ① Weigh out the following raw materials according to the proportions: 5-35 parts of Bupleurum chinense, 9-50 parts of Scutellaria baicalensis, 9-50 parts of Artemisia annua, 5-35 parts of Paeonia lactiflora, 5-35 parts of Paeonia suffruticosa, and 4-26 parts of Perilla frutescens.

[0051] ② Take Bupleurum chinense, Scutellaria baicalensis, and Artemisia annua, add 6 to 14 times the amount of water or ethanol, heat and reflux for 0.5 to 5 hours, heat and reflux for 1 to 3 times, filter, combine the extracts, and concentrate under reduced pressure at 40 to 90℃ to a density of 1.01 to 1.20 for later use;

[0052] ③ Take peony bark, red peony root, and perilla leaf, add 6-14 times the amount of raw herbs to 50%-90% ethanol or water, heat and reflux for 0.5-5 hours, heat and reflux for 1-3 times, filter, combine the extracts, concentrate under reduced pressure at 40-90℃ to a density of 1.01-1.20, combine the concentrate from step ②, and continue to concentrate to a density of 1.05-1.15, for later use;

[0053] ④ Dry the concentrated liquid from step ③ at 50–90°C, pulverize the dried paste, add excipients, and prepare the formulation.

[0054] To address the aforementioned technical problems, the present invention also provides a method for preparing a traditional Chinese medicine composition, the method comprising the following steps:

[0055] ① Weigh out the following raw materials according to the proportions: 5-35 parts of Bupleurum chinense, 9-50 parts of Scutellaria baicalensis, 9-50 parts of Artemisia annua, 5-35 parts of Paeonia lactiflora, 5-35 parts of Paeonia suffruticosa, and 4-26 parts of Perilla frutescens.

[0056] ② Take Bupleurum chinense, Scutellaria baicalensis, Artemisia annua, Paeonia lactiflora, Paeonia suffruticosa, and Perilla frutescens leaves, add 6-12 times the amount of raw herbs to 50%-90% ethanol, heat and reflux for 1-3 hours, heat and reflux for 1-3 times, filter, combine the extracts, and concentrate under reduced pressure at 50-90℃ to a density of 1.05-1.20 for later use;

[0057] ③Dry the concentrated liquid from step ② at 50-90℃, pulverize the dried paste, add excipients, and prepare the formulation.

[0058] To solve the above-mentioned technical problems, the present invention also provides a method for preparing a traditional Chinese medicine composition, the method comprising the following steps:

[0059] ① Weigh out the following raw materials according to the proportions: 5-35 parts of Bupleurum chinense, 9-50 parts of Scutellaria baicalensis, 9-50 parts of Artemisia annua, 5-35 parts of Paeonia lactiflora, 5-35 parts of Paeonia suffruticosa, and 4-26 parts of Perilla frutescens.

[0060] ② Take Bupleurum chinense, Scutellaria baicalensis, and Paeonia lactiflora, grind them into powder, sift them to obtain medium powder, and set aside;

[0061] ③ Take peony bark, perilla leaf, and artemisia annua, add 6 to 14 times the amount of raw herbs to water, soak for 0.5 to 1 hour, heat and reflux for 1 to 3 hours, heat and reflux for 1 to 3 times, filter, combine the extracts, concentrate under reduced pressure at 50 to 90℃ to a density of 1.05 to 1.20, add the powder from step ②, stir evenly, and set aside.

[0062] ④ Dry the thick paste from step ③ at 50–90°C, pulverize the dried paste, add excipients, and prepare the formulation.

[0063] To address the aforementioned technical problems, the present invention also provides a method for preparing a traditional Chinese medicine composition, the method comprising the following steps:

[0064] ① Weigh out the following raw materials according to the proportions: 5-35 parts of Bupleurum chinense, 9-50 parts of Scutellaria baicalensis, 9-50 parts of Artemisia annua, 5-35 parts of Paeonia lactiflora, 5-35 parts of Paeonia suffruticosa, and 4-26 parts of Perilla frutescens.

[0065] ② Take Bupleurum chinense, Scutellaria baicalensis, Artemisia annua, Paeonia lactiflora, Paeonia suffruticosa, and Perilla frutescens leaves, grind them into powder, sift them, and set aside;

[0066] ③ Add the powder from step ② to the excipients to prepare the formulation. Attached Figure Description

[0067] Figure 1 This is a WB diagram of HA protein expression in the composition of the present invention.

[0068] Figure 2 This is a WB diagram of NP protein expression in the composition of the present invention.

[0069] Figure 3 This is a PCR diagram of NP gene expression in the composition of the present invention.

[0070] Figure 4 This is a line graph comparing rectal temperature of the control drug in the antipyretic test with that of the composition of the present invention. Detailed Implementation

[0071] The following examples are used to illustrate the preparation of the drug of the present invention, but they should not be construed as limiting the scope of the present invention.

[0072] Example 1

[0073] The application of a traditional Chinese medicine composition in the preparation of a drug for treating colds and influenza caused by viruses, wherein the raw materials, by weight, include 30 parts of Bupleurum chinense, 40 parts of Scutellaria baicalensis, 40 parts of Artemisia annua, 20 parts of Paeonia lactiflora, 20 parts of Paeonia suffruticosa, and 10 parts of Perilla frutescens.

[0074] Specific preparation steps:

[0075] ① Take Bupleurum chinense, Scutellaria baicalensis, and Artemisia annua, add 12 times the amount of raw herbs to water, reflux and extract for 2 hours, filter, and set aside the filtrate. Add 12 times the amount of raw herbs to 80% ethanol, reflux and extract for 2 hours, filter, combine the two extracts, and concentrate under reduced pressure at 60℃ to a density of 1.08, and set aside.

[0076] ② Take peony bark, red peony root, and perilla leaf, add 12 times the amount of raw herbs to 80% ethanol, reflux extract for 2 hours, filter, and set aside the filtrate. Add 12 times the amount of raw herbs to the residue, reflux extract for 2 hours, filter, combine the two extracts, concentrate under reduced pressure at 60℃ to a density of 1.08, combine the concentrate from step ①, and continue to concentrate to a density of 1.10, and set aside.

[0077] ③ The concentrate from step ② is dried at 70°C, the dried paste is pulverized and excipients are added to prepare capsules, thus obtaining the invention composition 1.

[0078] Example 2

[0079] The application of a traditional Chinese medicine composition in the preparation of a drug for treating colds and influenza caused by viruses, wherein the raw materials, by weight, include 25 parts of Bupleurum chinense, 35 parts of Scutellaria baicalensis, 40 parts of Artemisia annua, 15 parts of Paeonia lactiflora, 15 parts of Paeonia suffruticosa, and 18 parts of Perilla frutescens.

[0080] Specific preparation steps:

[0081] ① Take Bupleurum chinense, Scutellaria baicalensis, and Artemisia annua, add 12 times the amount of raw herbs to water, reflux and extract for 2 hours, filter, and set aside the filtrate. Add 12 times the amount of raw herbs to 80% ethanol, reflux and extract for 2 hours, filter, combine the two extracts, and concentrate under reduced pressure at 60℃ to a density of 1.08, and set aside.

[0082] ② Take peony bark, red peony root, and perilla leaf, add 12 times the amount of raw herbs to 80% ethanol, reflux extract for 2 hours, filter, and set aside the filtrate. Add 12 times the amount of raw herbs to the residue, reflux extract for 2 hours, filter, combine the two extracts, concentrate under reduced pressure at 60℃ to a density of 1.08, combine the concentrate from step ①, and continue to concentrate to a density of 1.10, and set aside.

[0083] ③ The concentrate from step ② is dried at 70°C, the dried paste is pulverized and then excipients are added to prepare granules, thus obtaining the invention composition 2.

[0084] Example 3

[0085] The application of a traditional Chinese medicine composition in the preparation of a drug for treating colds and influenza caused by viruses, wherein the raw materials, by weight, include 20 parts of Bupleurum chinense, 30 parts of Scutellaria baicalensis, 30 parts of Artemisia annua, 20 parts of Paeonia lactiflora, 20 parts of Paeonia suffruticosa, and 18 parts of Perilla frutescens.

[0086] Specific preparation steps:

[0087] ① Take Bupleurum chinense, Scutellaria baicalensis, Artemisia annua, Paeonia lactiflora, Paeonia suffruticosa, and Perilla frutescens leaves, add 12 times the amount of raw herbs to water, reflux and extract for 2 hours, filter, and set aside the filtrate. Add 12 times the amount of raw herbs to the residue, reflux and extract for 2 hours, filter, combine the two extracts, and concentrate under reduced pressure at 60℃ to a density of 1.08, and set aside.

[0088] ② The concentrate is dried at 60°C, the dried paste is pulverized and excipients are added to prepare capsules, thus obtaining the invention composition 3.

[0089] Example 4

[0090] The application of a traditional Chinese medicine composition in the preparation of a drug for treating colds and influenza caused by viruses, wherein the raw materials, by weight, include 15 parts of Bupleurum chinense, 35 parts of Scutellaria baicalensis, 30 parts of Artemisia annua, 30 parts of Paeonia lactiflora, 20 parts of Paeonia suffruticosa, and 10 parts of Perilla frutescens.

[0091] Specific preparation steps:

[0092] ① Take Bupleurum chinense, Scutellaria baicalensis, Artemisia annua, Paeonia lactiflora, Paeonia suffruticosa, and Perilla frutescens leaves, add 12 times the amount of raw herbs to water, reflux and extract for 2 hours, filter, and set aside the filtrate. Add 12 times the amount of raw herbs to the residue, reflux and extract for 2 hours, filter, combine the two extracts, and concentrate under reduced pressure at 60℃ to a density of 1.08, and set aside.

[0093] ② The concentrate is dried at 60℃, the dried paste is pulverized and then excipients are added to prepare granules.

[0094] Example 5

[0095] The application of a traditional Chinese medicine composition in the preparation of a drug for treating colds and influenza caused by viruses, wherein the raw materials, by weight, include 18 parts of Bupleurum chinense, 30 parts of Scutellaria baicalensis, 20 parts of Artemisia annua, 20 parts of Paeonia lactiflora, 30 parts of Paeonia suffruticosa, and 15 parts of Perilla frutescens.

[0096] Specific preparation steps:

[0097] ① Take Bupleurum chinense, Scutellaria baicalensis, and Artemisia annua, add 10 times the amount of raw herbs to 60% ethanol, reflux extract for 1.5 hours, filter, and set aside the filtrate. Add 10 times the amount of raw herbs to 60% ethanol, reflux extract for 1.5 hours, filter, combine the two extracts, and concentrate under reduced pressure at 60℃ to a density of 1.06, and set aside.

[0098] ② Take peony bark, red peony root, and perilla leaf, add 10 times the amount of raw herbs to water, reflux and extract for 1.5 hours, filter, and set aside the filtrate. Add 10 times the amount of raw herbs to the residue, reflux and extract for 1.5 hours, filter, combine the two extracts, and concentrate under reduced pressure at 70℃ to a density of 1.06. Combine the concentrate from step ① and continue to concentrate to a density of 1.09, and set aside.

[0099] ③Dry the concentrated liquid from step ② at 60℃, pulverize the dried paste, add excipients, and prepare capsules.

[0100] Example 6

[0101] The application of a traditional Chinese medicine composition in the preparation of a drug for treating colds and influenza caused by viruses, wherein the raw materials, by weight, include 15 parts of Bupleurum chinense, 25 parts of Scutellaria baicalensis, 35 parts of Artemisia annua, 15 parts of Paeonia lactiflora, 20 parts of Paeonia suffruticosa, and 20 parts of Perilla frutescens.

[0102] Specific preparation steps:

[0103] ① Take Bupleurum chinense, Scutellaria baicalensis, and Artemisia annua, add 10 times the amount of raw herbs to 60% ethanol, reflux extract for 1.5 hours, filter, and set aside the filtrate. Add 10 times the amount of raw herbs to 60% ethanol, reflux extract for 1.5 hours, filter, combine the two extracts, and concentrate under reduced pressure at 60℃ to a density of 1.06, and set aside.

[0104] ② Take peony bark, red peony root, and perilla leaf, add 10 times the amount of raw herbs to water, reflux and extract for 1.5 hours, filter, and set aside the filtrate. Add 10 times the amount of raw herbs to the residue, reflux and extract for 1.5 hours, filter, combine the two extracts, and concentrate under reduced pressure at 70℃ to a density of 1.06. Combine the concentrate from step ① and continue to concentrate to a density of 1.09, and set aside.

[0105] ③Dry the concentrated liquid from step ② at 60℃, pulverize the dried paste, add excipients, and prepare granules.

[0106] Example 7

[0107] The application of a traditional Chinese medicine composition in the preparation of a drug for treating colds and influenza caused by viruses, wherein the raw materials, by weight, include 30 parts of Bupleurum chinense, 30 parts of Scutellaria baicalensis, 30 parts of Artemisia annua, 25 parts of Paeonia lactiflora, 25 parts of Paeonia suffruticosa, and 15 parts of Perilla frutescens.

[0108] Specific preparation steps:

[0109] ① Take Bupleurum chinense, Scutellaria baicalensis, and Artemisia annua, add 10 times the amount of raw herbs to 50% ethanol, reflux extract for 1.5 hours, filter, and set aside the filtrate. Add 10 times the amount of raw herbs to 50% ethanol, reflux extract for 1 hour, filter, combine the two extracts, and concentrate under reduced pressure at 60℃ to a density of 1.05, and set aside.

[0110] ② Take peony bark, red peony root, and perilla leaf, add 10 times the amount of raw herbs to water, reflux and extract for 1.5 hours, filter, and set aside the filtrate. Add 10 times the amount of raw herbs to the residue, reflux and extract for 1 hour, filter, combine the two extracts, and concentrate under reduced pressure at 60℃ to a density of 1.04. Combine the concentrate from step ① and continue to concentrate to a density of 1.09, and set aside.

[0111] ③Dry the concentrated liquid from step ② at 60℃, pulverize the dried paste, add excipients, and prepare capsules.

[0112] Example 8

[0113] The application of a traditional Chinese medicine composition in the preparation of a drug for treating colds and influenza caused by viruses, wherein the raw materials, by weight, include 20 parts of Bupleurum chinense, 30 parts of Scutellaria baicalensis, 30 parts of Artemisia annua, 20 parts of Paeonia lactiflora, 20 parts of Paeonia suffruticosa, and 15 parts of Perilla frutescens.

[0114] Specific preparation steps:

[0115] ① Take Bupleurum chinense, Scutellaria baicalensis, and Artemisia annua, add 10 times the amount of raw herbs to 70% ethanol, reflux extract for 1.5 hours, filter, and set aside the filtrate. Add 10 times the amount of raw herbs to 70% ethanol, reflux extract for 1 hour, filter, combine the two extracts, and concentrate under reduced pressure at 60℃ to a density of 1.05, and set aside.

[0116] ② Take peony bark, red peony root, and perilla leaf, add 10 times the amount of raw herbs to water, reflux and extract for 1.5 hours, filter, and set aside the filtrate. Add 10 times the amount of raw herbs to the residue, reflux and extract for 1 hour, filter, combine the two extracts, and concentrate under reduced pressure at 60℃ to a density of 1.04. Combine the concentrate from step ① and continue to concentrate to a density of 1.09, and set aside.

[0117] ③Dry the concentrated liquid from step ② at 60℃, pulverize the dried paste, add excipients, and prepare granules.

[0118] Example 9

[0119] The application of a traditional Chinese medicine composition in the preparation of a medicine for treating colds and influenza caused by viruses, wherein the raw materials, by weight, include 30 parts of Bupleurum chinense, 40 parts of Scutellaria baicalensis, 40 parts of Artemisia annua, 30 parts of Paeonia lactiflora, 30 parts of Paeonia suffruticosa, and 20 parts of Perilla frutescens.

[0120] Specific preparation steps:

[0121] ① Take Bupleurum chinense, Scutellaria baicalensis, Artemisia annua, Paeonia lactiflora, Paeonia suffruticosa, and Perilla frutescens leaves, add 10 times the amount of raw herbs to 80% ethanol, reflux extract for 1.5 hours, filter, and set aside the filtrate. Add 10 times the amount of raw herbs to the residue to 80% ethanol, reflux extract for 1.5 hours, filter, combine the two extracts, and concentrate under reduced pressure at 60℃ to a density of 1.10, and set aside.

[0122] ② The concentrated liquid from step ① is dried at 60℃, the dried paste is pulverized and excipients are added to prepare capsules.

[0123] Example 10

[0124] The application of a traditional Chinese medicine composition in the preparation of a drug for treating colds and influenza caused by viruses, wherein the raw materials, by weight, include 32 parts of Bupleurum chinense, 48 parts of Scutellaria baicalensis, 45 parts of Artemisia annua, 28 parts of Paeonia lactiflora, 28 parts of Paeonia suffruticosa, and 18 parts of Perilla frutescens.

[0125] Specific preparation steps:

[0126] ① Take Bupleurum chinense, Scutellaria baicalensis, Artemisia annua, Paeonia lactiflora, Paeonia suffruticosa, and Perilla frutescens leaves, grind them into powder, sift them, and set aside;

[0127] ② Add the powder from step ① to excipients to prepare capsules.

[0128] Example 11

[0129] The application of a traditional Chinese medicine composition in the preparation of a drug for treating colds and influenza caused by viruses, wherein the raw materials, by weight, include 20 parts of Bupleurum chinense, 30 parts of Scutellaria baicalensis, 40 parts of Artemisia annua, 15 parts of Paeonia lactiflora, 18 parts of Paeonia suffruticosa, and 12 parts of Perilla frutescens.

[0130] Specific preparation steps:

[0131] ① Take Bupleurum chinense, Scutellaria baicalensis, and Paeonia lactiflora, grind them into powder, sift them to obtain medium powder, and set aside;

[0132] ② Take peony bark, perilla leaf, and artemisia annua, add 6-14 times the amount of raw herbs to water, soak for 0.5-1 hour, heat and reflux for 1-3 hours, heat and reflux for 1-3 times, filter, combine the extracts, concentrate under reduced pressure at 50-90℃ to a density of 1.05-1.20, add the powder from step ①, stir evenly, and set aside.

[0133] ③Dry the thick paste from step ② at 60℃, pulverize the dried paste, add excipients, and prepare granules.

[0134] Comparative Example 1

[0135] The traditional Chinese medicine composition comprises, by weight, 30 parts of Bupleurum chinense, 40 parts of Scutellaria baicalensis, 40 parts of Artemisia annua, 40 parts of Isatis tinctoria, 30 parts of Pueraria lobata, 30 parts of Paeonia lactiflora, 30 parts of Paeonia suffruticosa, 20 parts of Ligusticum chuanxiong, and 20 parts of Perilla frutescens.

[0136] Specific preparation steps:

[0137] ① Take Bupleurum chinense, Scutellaria baicalensis, Artemisia annua, Isatis tinctoria, and Ligusticum chuanxiong. Add 10 times the amount of raw herbs in 70% ethanol and reflux for 1.5 hours. Filter and set aside the filtrate. Add 10 times the amount of raw herbs in 70% ethanol to the residue and reflux for 1 hour. Filter and combine the two extracts. Concentrate under reduced pressure at 60℃ to a density of 1.05 and set aside.

[0138] ② Take kudzu root, peony bark, red peony root, and perilla leaf, add 10 times the amount of raw herbs to water, reflux and extract for 1.5 hours, filter, and set aside the filtrate. Add 10 times the amount of raw herbs to the residue, reflux and extract for 1 hour, filter, combine the two extracts, and concentrate under reduced pressure at 60℃ to a density of 1.04. Combine the concentrate from step ① and continue to concentrate to a density of 1.09, and set aside.

[0139] ③ The concentrate from step ② is dried at 60°C, the dried paste is pulverized and then excipients are added to prepare granules, thus obtaining comparative composition 1.

[0140] Comparative Example 2

[0141] The traditional Chinese medicine composition comprises, by weight, 40 parts of Scutellaria baicalensis, 40 parts of Artemisia annua, 30 parts of Paeonia suffruticosa, and 20 parts of Perilla frutescens.

[0142] Specific preparation steps:

[0143] ① Take Scutellaria baicalensis and Artemisia annua, add 10 times the amount of raw herbs in 70% ethanol, reflux extract for 1.5 hours, filter, and set aside the filtrate. Add 10 times the amount of raw herbs in 70% ethanol to the residue, reflux extract for 1 hour, filter, combine the two extracts, and concentrate under reduced pressure at 60℃ to a density of 1.05, and set aside.

[0144] ② Take peony bark and perilla leaves, add 10 times the amount of raw herbs to water, reflux and extract for 1.5 hours, filter, and set aside the filtrate. Add 10 times the amount of raw herbs to the residue, reflux and extract for 1 hour, filter, combine the two extracts, concentrate under reduced pressure at 60℃ to a density of 1.04, combine the concentrate from step ①, and continue to concentrate to a density of 1.09, and set aside.

[0145] ③ The concentrate from step ② is dried at 60°C, the dried paste is pulverized and then excipients are added to prepare granules, thus obtaining comparative composition 2.

[0146] Comparative Example 3

[0147] The traditional Chinese medicine composition comprises, by weight, 30 parts of Bupleurum chinense, 40 parts of Scutellaria baicalensis, 40 parts of Isatis indigotica, 30 parts of Pueraria lobata, 30 parts of Paeonia lactiflora, 30 parts of Paeonia suffruticosa, 20 parts of Ligusticum chuanxiong, and 20 parts of Perilla frutescens.

[0148] Specific preparation steps:

[0149] ① Take Bupleurum chinense, Scutellaria baicalensis, Isatis indigotica, and Ligusticum chuanxiong, add 10 times the amount of raw herbs in 70% ethanol, reflux extract for 1.5 hours, filter, and set aside the filtrate. Add 10 times the amount of raw herbs in 70% ethanol to the residue, reflux extract for 1 hour, filter, combine the two extracts, and concentrate under reduced pressure at 60℃ to a density of 1.05, and set aside.

[0150] ② Take peony bark, red peony root, and perilla leaf, add 10 times the amount of raw herbs to water, reflux and extract for 1.5 hours, filter, and set aside the filtrate. Add 10 times the amount of raw herbs to the residue, reflux and extract for 1 hour, filter, combine the two extracts, and concentrate under reduced pressure at 60℃ to a density of 1.04. Combine the concentrate from step ① and continue to concentrate to a density of 1.09, and set aside.

[0151] ③ The concentrate from step ② is dried at 60°C, the dried paste is pulverized and then excipients are added to prepare granules, thus obtaining comparative composition 3.

[0152] Pharmacodynamic study of the traditional Chinese medicine composition of this invention:

[0153] Antiviral efficacy test of the traditional Chinese medicine composition of this invention:

[0154] 1. Materials

[0155] 1.1 Drugs and Reagents

[0156] Test drugs: (Inventive composition 1, Inventive composition 2, Comparative composition 1, Comparative composition 2)

[0157] Reagents:

[0158] RPMI 1640 medium (powder), Gibco;

[0159] Fetal bovine serum (FBS), BioInd;

[0160] WBRIPA cell lysis buffer, Beyotime Biotechnology Co., Ltd.

[0161] Phosphatase inhibitor, MCE Corporation;

[0162] Protease inhibitor mixture, Beijing Solarbio Science & Technology Co., Ltd.

[0163] Skim milk powder, Yili Industrial Group Co., Ltd.

[0164] Prestained Protein Maker, Thermo Fisher Scientific, Inc.

[0165] ECL luminescent fluid, Perkin Elmer;

[0166] PVDF membrane, Millipore;

[0167] DEPC, Sigma-Aldrich, Inc. (USA)

[0168] TRNzol Universal Reagent, Tiangen Biotech (Beijing) Co., Ltd.;

[0169] Isopropanol, Tianjin Bodi Chemical Co., Ltd.;

[0170] Chloroform, Tianjin Beifang Tianyi Chemical Reagent Factory;

[0171] PrimeScript™ RT reagent Kit with gDNA Eraser, TaKaRa Corporation, Japan;

[0172] PowerUPTMSYBRTM Green Master Mix, Applied Biosystems;

[0173] Anhydrous ethanol, Tianjin Yongda Chemical Reagent Co., Ltd.

[0174] Rabbit anti-GAPDH primary antibody, Wuhan Sanying Biotechnology Co., Ltd.

[0175] Mouse anti-NP primary antibody, Beijing Yiqiao Shenzhou Technology Co., Ltd.;

[0176] Mouse anti-HA primary antibody, Beijing Yiqiao Shenzhou Technology Co., Ltd.;

[0177] Primers, Sangon Biotech (Shanghai) Co., Ltd.;

[0178] 1.2 Cell types:

[0179] A549 cells, virus strains: H1N1, H3N2

[0180] instrument:

[0181] Electric thermostatic CO2 cell incubator, Thermo Scientific, USA;

[0182] Pipettes, Eppendorf GmbH, Germany;

[0183] Electronic balance, Ohaus Instruments (Changzhou) Co., Ltd.;

[0184] Ultrasonic crusher (VCX800 model), SONICS, USA;

[0185] Tabletop high-speed refrigerated centrifuge (model 23R), Shanghai Lishen Company;

[0186] 7500 Real-Time PCR System, Applied Biosystems, USA;

[0187] PCR thermal cycler (S1000TM model), BIO-RAD Corporation, USA;

[0188] Protein blotting device, BIO-RAD Corporation, USA;

[0189] Basic electrophoresis apparatus power supply, BIO-RAD Corporation, USA;

[0190] ECL chemiluminescence imaging system, Syngene Corporation, USA;

[0191] 2. Method:

[0192] Using A549 cells as host cells, after 2 hours of viral infection (to complete the adsorption process), the culture medium was replaced with a drug-containing medium (final concentration of 2.5 g / L). After 36 hours of treatment, the antiviral effect was measured.

[0193] 2.1 Viral infection of A549 cells

[0194] ① The day before infection, A549 was inoculated into a 12-well plate and incubated overnight at 37°C in a 5% CO2 incubator.

[0195] ② Discard the supernatant, wash the cells 1-2 times with PBS, add 1 mL of RPMI 1640 containing 2% FBS and double antibodies to each well, add the virus to the cells, and incubate at 34℃ and 5% CO2 for 2-3 hours, shaking the plate every 15-20 minutes during this period to ensure full infection;

[0196] ③ After adsorption is complete, add 1 mL of RPMI 1640 containing 2% FBS and double antibiotics to each well, and continue to incubate in a constant temperature incubator of 34℃ and 5% CO2 for 36 h;

[0197] ④ After culturing for 36 hours, the culture medium was replaced with RPMI 1640 containing 2% FBS, double antibiotics and traditional Chinese medicine extracts (Inventory Composition 1, Inventory Composition 2, Comparative Example 1, Comparative Example 2), and cultured in a constant temperature incubator at 34℃ until the detection time point.

[0198] 2.2 Grouping and Dosing

[0199] Weigh 0.625g of the extract, dissolve it in 250ml of 2% 1640 medium, filter it through a 0.45um filter membrane, and add it directly to the cells. The final drug concentration is 2.5g / L.

[0200] Western blot was used to detect GAPD and NP proteins. qRT-PCR was used to detect the NP gene and ACTIN.

[0201] 2.3 Experimental results are attached to the instruction manual. Figure 1 , Figure 2 and Figure 3

[0202] Conclusion 1: All four formulations can significantly inhibit H3N2 and HA protein expression levels, suggesting that the four traditional Chinese medicines can inhibit H3N2 replication. Among them, the inhibitory effects of Invented Composition 1 and Invented Composition 2 are the most significant, and the antiviral effects are the best.

[0203] Conclusion 2: Composition 1 and Composition 2 of the present invention can significantly inhibit the expression levels of H1N1 NP gene and protein, suggesting that the four traditional Chinese medicines can inhibit H1N1 replication.

[0204] Antipyretic test of the traditional Chinese medicine composition of this invention:

[0205] 1. Test Methods

[0206] 1.1 Adaptation period: SPF-grade male SD rats, weighing 160-180g, were acclimatized for 4 days and their rectal temperature was measured twice a day for 3 days.

[0207] 1.2 Before the experiment: Fasting was allowed for 6 hours before the experiment, but water was allowed. Rectal temperature was measured 3 times (1 time / 30min) before the experiment and the average body temperature was calculated. The group was enrolled if △T≤0.5℃ and 36.6~38.4℃.

[0208] 1.3 Modeling: 20% yeast (10mg / kg) was injected subcutaneously into the back. For the first 4 hours after modeling, the temperature was measured every 30 minutes for a total of 8 times. Patients whose body temperature rose by more than 0.8℃ were selected for the group.

[0209] 1.4 Gavage: After successful modeling, rats were randomly divided into four groups of 50 (n=10 per group). The groups included a model group, a positive control group, a low-dose administration group (0.5 times the human dose), a medium-dose administration group (human dose), and a high-dose administration group (twice the human dose). The model group received the corresponding volume of physiological saline, the positive control group received the corresponding volume of Qingkailing granules and comparative composition 1 and comparative composition 2, and the administration groups received the corresponding concentration of composition 1 of this invention. Administration was performed three times daily via gavage, with rectal temperature measured every 2 hours until 24 hours after administration, for a total of 12 times.

[0210] 1.5 Data Processing: Experimental data were expressed as mean values. SPSS 27.0 software was used for statistical analysis of the experimental data. One-way ANOVA was used for comparisons among multiple groups. A p-value less than 0.05 was considered statistically significant.

[0211] 2. The test results are shown in Tables 1 and 2, and are attached to the instruction manual. Figure 4 .

[0212] Table 1. Results of changes in body temperature over time

[0213] 0h 2h 4h 6h 8h 10h 12h 14h 16h 18h 20h 22h 24h Model 39.4 39.3 38.9 39.2 39.0 38.8 38.8 38.7 39.0 38.9 38.8 38.8 38.7 Positive control 39.2 39.3 38.8 38.7 38.8 38.8 38.7 38.5 38.5 38.3 38.1 38.3 38.1 low dose 39.2 39.4 39.0 38.9 38.5 38.9 38.8 38.7 38.6 38.4 38.1 38.3 38.1 medium dose 39.5 39.0 38.9 38.5 38.0 38.1 38.0 38.0 38.2 38.2 38.3 38.0 37.5 High dose 39.8 39.1 39.2 38.5 38.2 37.9 38.1 38.0 38.3 38.1 38.1 37.9 37.6 Comparative Example 1 39.6 39.7 39.3 39.1 39.1 38.8 38.9 38.7 38.6 38.4 38.5 38.4 38.4 Comparative Example 2 39.3 39.6 39.3 39.2 38.6 38.9 38.8 38.6 38.6 38.4 38.3 38.3 38.3

[0214] Table 2. Results of P-value variation over time

[0215] p-value 0h 2h 4h 6h 8h 10h 12h 14h 16h 18h 20h 22h 24h Model / Positive 0.537 1.000 0.712 0.215 0.264 0.788 0.260 0.258 0.074 0.138 0.050 0.048 0.014 Model / Low 0.539 0.535 0.902 0.133 0.140 0.711 0.082 0.610 0.086 0.133 0.086 0.064 0.014 Model / Middle 0.455 0.211 1.000 0.014 0.013 0.014 0.014 0.014 0.014 0.033 0.049 0.012 0.004 Model / High 0.108 0.618 0.108 0.018 0.014 0.014 0.036 0.014 0.014 0.035 0.019 0.014 0.009 Model / Comparative Example 1 0.543 0.189 0.097 0.723 0.689 0.788 0.356 0.611 0.086 0.133 0.137 0.106 0.218 Model / Comparative Example 2 0.457 0.215 0.097 1.000 0.188 0.711 0.585 0.476 0.094 0.133 0.052 0.059 0.176 Positive / Low 0.616 0.916 0.343 0.671 0.207 0.429 0.390 0.334 0.595 0.750 0.753 0.459 0.915 Positive / Medium 0.415 0.248 0.832 0.465 0.008 0.007 0.009 0.011 0.344 0.527 0.249 0.130 0.034 Positive / High 0.214 0.459 0.059 0.401 0.012 0.007 0.012 0.011 0.596 0.596 0.833 0.141 0.008 Positive / Positive Example 1 0.495 0.154 0.047 0.201 0.258 0.908 0.696 0.334 0.595 0.747 0.111 0.325 0.269 Positive / Positive Ratio 2 0.388 0.266 0.048 0.135 0.199 0.776 0.893 0.432 0.564 0.761 0.248 0.461 0.313

[0216] 3. Experimental conclusions: Compared with the rectal temperature of the model group rats, the antipyretic effects of the positive control group and the low-dose group of the composition of the present invention were comparable, while the antipyretic effects of the medium-dose group and the high-dose group of the composition of the present invention were comparable. After 3 oral boluses and 24 hours later, the body temperature showed a significant downward trend. The experimental data show that the traditional Chinese medicine composition of the present invention can play a significant antipyretic role.

[0217] Typical clinical applications of the traditional Chinese medicine composition of this invention:

[0218] Case 1: Ms. Feng, female, 73 years old. July 9, 2024. Visited and was hospitalized at Ping An Hospital.

[0219] Chief complaint: Nasal congestion, runny nose, sneezing, severe sore throat for 4 days, fatigue, accompanied by fever for 2 days, with a highest temperature of 38.4 degrees Celsius. Physical examination: The patient is alert and speaks fluently, with fair mental status. Pharyngeal congestion, slightly coarse breath sounds in both lungs, no abnormalities in the heart or abdomen, and no edema in the lower extremities.

[0220]

[0221] Diagnosis: Wind-heat. Treatment principle: Clear heat and detoxify, expel pathogens and release the exterior.

[0222] The patient took 2 granules of the composition of this invention for 3 days, twice a day, 15g each time. On February 22, the body temperature was 37.2-37.5 degrees Celsius, and the sore throat and runny nose improved. The patient continued to take the granules for 5 days, twice a day, 10g each time. The body temperature returned to normal, all symptoms improved, and the patient was discharged from the hospital.

[0223] Case 2: Kang, female, 27 years old, visited Ping An Hospital on June 5, 2025.

[0224] Chief complaint: Nasal congestion, runny nose, and severe dry and sore throat for one day, accompanied by a mild cough. Fever of 38.4°C.

[0225] Physical examination: The patient was alert and articulate, with fair mental status. There was significant pharyngeal congestion, slightly coarse breath sounds in both lungs, negative findings in the heart and abdomen, and no edema in the lower extremities.

[0226]

[0227] Diagnosis: Wind-heat. Treatment principle: Clear heat and detoxify, expel pathogens and release the exterior.

[0228] The patient took 2 granules of the composition of this invention for 3 days, twice a day, 15g each time. On June 8th, the body temperature was 37.2-37.5 degrees Celsius, and the sore throat and cough improved. The patient continued to take the granules for 5 days, twice a day, 10g each time. Follow-up visit showed that the body temperature was normal and all symptoms had improved.

[0229] Case 3: Hou, male, 68 years old. He visited Ping An Hospital on May 9, 2024.

[0230] The patient complained of dry throat, sore throat, and fatigue. Occasional cough, but no sputum, chest tightness, or shortness of breath. Physical examination revealed a pale tongue with a white coating, a weak pulse, blood pressure of 150 / 89 mmHg, no facial edema, pharyngeal congestion, no tonsillar enlargement, coarse breath sounds in both lungs without any dry or wet rales, a heart rate of 90 beats / min with a regular rhythm, and no murmurs heard in any of the valvular auscultation areas. The patient also had a fever, with a temperature of 38.3℃.

[0231]

[0232] Diagnosis: Wind-heat. Treatment principle: Clear heat and detoxify, expel pathogens and release the exterior.

[0233] The patient took 2 granules of the composition of this invention for 7 days, twice a day, 10g of granules each time. A follow-up visit on July 16, 2024, showed normal body temperature and improved symptoms.

[0234] Case 4: Chen, female, 76 years old, was admitted to Ping An Hospital on March 12, 2025.

[0235] The patient presented with intermittent fever, poor appetite, nausea, vomiting, cough, sputum production, nasal congestion, and elevated body temperature, reaching a maximum of 38.2℃. The tongue coating was thin and white, and the pulse was wiry.

[0236]

[0237] Diagnosis: Wind-heat syndrome. Differential diagnosis: Clear heat and detoxify, expel pathogens and release the exterior.

[0238] The patient took two granules of the composition of this invention twice a day, 20g each time. Two hours after administration, the body temperature began to decrease, and returned to normal three days later, on March 15th. Nasal congestion and cough with phlegm improved.

[0239] Continue taking the compound granules twice a day, 10g each time. On March 18, all symptoms improved and the patient was discharged with a normal body temperature.

[0240] Case 5: Wang, male, 59 years old. He visited Ping An Hospital on December 5, 2024.

[0241] The patient's mental state, appetite, and sleep are normal. They experience intermittent nasal congestion, runny nose, sore throat, cough, and fatigue. Their body temperature is 38.5℃.

[0242] Physical examination: Red tongue with thin yellow coating, wiry pulse.

[0243]

[0244]

[0245] Diagnosis: Wind-Heat Syndrome. Treatment Principle: Clear heat and detoxify, expel pathogens and release the exterior.

[0246] The patient took two granules of the composition of this invention twice a day, 20g each time. After taking the medication, the body temperature began to decrease after 4 hours. On the morning of December 6th, the body temperature was 37.8℃. Two days later, the body temperature returned to normal, below 37.2℃, and the nasal congestion and cough improved.

[0247] Continue taking the compound granules twice a day, 10g each time. A follow-up visit on December 20th showed improvement in all symptoms and normal body temperature.

Claims

1. The application of a traditional Chinese medicine composition in the preparation of medicines for treating colds and influenza caused by viruses, characterized in that, The weight ratio of the Chinese medicinal materials is as follows: Bupleurum chinense 5-35 parts, Scutellaria baicalensis 9-50 parts, Artemisia annua 9-50 parts, Paeonia lactiflora 5-35 parts, Paeonia suffruticosa 5-35 parts, and Perilla frutescens 4-26 parts.

2. The application according to claim 1, characterized in that, The weight ratio of the Chinese medicinal materials is as follows: Bupleurum chinense 8-28 parts, Scutellaria baicalensis 12-40 parts, Artemisia annua 12-40 parts, Paeonia lactiflora 8-28 parts, Paeonia suffruticosa 8-28 parts, and Perilla frutescens 8-22 parts.

3. The application according to claim 1, characterized in that, The weight ratio of the Chinese medicinal materials is as follows: Bupleurum chinense 10-25 parts, Scutellaria baicalensis 20-35 parts, Artemisia annua 20-35 parts, Paeonia lactiflora 10-25 parts, Paeonia suffruticosa 10-25 parts, and Perilla frutescens 12-20 parts.

4. The application according to any one of claims 1 to 3, characterized in that, The traditional Chinese medicine composition may be prepared into any commonly used preparation, such preparation as tablets, capsules, granules, pills, mixtures, syrups, decoctions, powders, suppositories, gels, sprays, or injections.

Images