A traditional Chinese medicine composition for clearing heat, dispersing stagnation, and detoxifying: its preparation, detection, and application.

Abstract

This invention discloses a traditional Chinese medicine composition for clearing heat and detoxifying, its preparation, detection, and application. The composition uses honeysuckle, forsythia, mulberry leaf, and chrysanthemum as the principal herbs; ephedra, bitter almond, platycodon, and bupleurum as the assistant herbs; and reed rhizome, loquat leaf, verbena, and codonopsis as adjuvant herbs. Licorice harmonizes the other herbs. Together, these herbs work to dispel wind and clear heat, promote lung function and relieve cough, and strengthen the body's resistance to pathogens. The extract of the above-mentioned traditional Chinese medicine composition is further prepared into a pharmaceutical composition using a pharmaceutically acceptable carrier. By analyzing the relationship between "drug-component-intersecting target," the mechanism of action between the above-mentioned traditional Chinese medicine composition and the common cold virus and the novel coronavirus was revealed. Simultaneously, a comprehensive quality control method was established to ensure that the traditional Chinese medicine composition achieves definite therapeutic effects.

Description

Technical Field

[0001] This invention relates to a traditional Chinese medicine composition for clearing heat and detoxifying, its preparation, detection, and application, specifically to a traditional Chinese medicine composition for clearing heat and wind, relieving cough and cough, and strengthening the body's resistance to pathogens, its preparation, detection, and application. Background Technology

[0002] Upper respiratory tract infection (URTI) refers to a general term for acute infectious inflammation of the pharynx, larynx, or nasal cavity. It is one of the most common infectious diseases of the respiratory tract, with 70-80% caused by viruses, mainly respiratory syncytial virus (RSV), rhinovirus, adenovirus, influenza viruses (A, B, and C), parainfluenza virus, and coronaviruses. Clinical manifestations include varying degrees of chills, fever, sore throat, nasal congestion, runny nose, sneezing, cough, fatigue, general muscle aches, pharyngeal congestion, and tonsillar enlargement. Influenza, an acute respiratory infection caused by the influenza virus, is also a highly contagious and rapidly spreading disease, mainly transmitted through airborne droplets, person-to-person contact, or contact with contaminated objects. Typical clinical symptoms include: sudden onset of high fever, body aches, significant fatigue, and mild respiratory symptoms. COVID-19 is a respiratory infection caused by a coronavirus. The main transmission routes are respiratory droplets and contact. Common signs of coronavirus infection include respiratory symptoms such as fever, cough, shortness of breath, and difficulty breathing. In more severe cases, infection can lead to pneumonia, severe acute respiratory syndrome, kidney failure, and even death. More than 95% of cases of the Omega-3 variant of COVID-19 are mild or asymptomatic. The clinical manifestations of mild COVID-19 are mainly due to external wind-cold, such as sore throat, cough, fever, nasal congestion, runny nose, and fatigue, with a red tongue, thin yellow coating, and floating pulse. Influenza and COVID-19 both belong to the category of upper respiratory tract infections.

[0003] Currently, Western medicine treats viral upper respiratory tract infections primarily with broad-spectrum antibiotics and comprehensive symptomatic treatment to relieve symptoms. However, the treatment effect is not ideal, long-term use has significant side effects, and it cannot cure the symptoms. Summary of the Invention

[0004] Purpose of the invention: In order to overcome the shortcomings of existing technologies, such as poor efficacy and inability to cure the disease, this invention aims to provide a traditional Chinese medicine composition and its composition with the effects of dispelling wind and clearing heat, ventilating the lungs and relieving cough, and supporting the body's resistance and eliminating pathogens. It also provides their preparation methods, detection methods and applications.

[0005] Technical solution: The traditional Chinese medicine composition for clearing heat and detoxifying described in this invention comprises, by weight, 6-12 parts of mulberry leaf, 6-12 parts of chrysanthemum, 6-12 parts of forsythia, 6-12 parts of honeysuckle, 3-7 parts of platycodon, 6-12 parts of bitter almond, 6-12 parts of honey loquat leaf, 2-5 parts of ephedra, 4-9 parts of bupleurum, 9-14 parts of verbena, 12-16 parts of reed rhizome, 6-12 parts of codonopsis, and 3-7 parts of licorice.

[0006] Furthermore, by weight, the ratio of mulberry leaves, chrysanthemum, forsythia, and honeysuckle is 1:1:1:1.

[0007] Furthermore, by weight, the above-mentioned traditional Chinese medicine composition comprises 10 parts of mulberry leaf, 10 parts of chrysanthemum, 10 parts of forsythia, 10 parts of honeysuckle, 6 parts of platycodon, 10 parts of bitter almond, 10 parts of honey loquat leaf, 3 parts of ephedra, 8 parts of bupleurum, 12 parts of verbena, 15 parts of reed rhizome, 10 parts of codonopsis, and 6 parts of licorice.

[0008] Furthermore, in the above-mentioned traditional Chinese medicine composition, the ephedra is raw ephedra and the licorice is raw licorice.

[0009] Traditional Chinese medicine (TCM) considers viral colds to be generally referred to as "common colds" or "wind-colds." Most colds are caused by insufficient vital energy (Qi) combined with external pathogenic factors (wind-evil) that invade the body. The primary location of the disease is in the lungs and wei qi (defensive Qi). The lungs govern the skin and hair; their dispersing function ensures that the body's essential substances reach the skin and hair. Since the skin and hair are the outermost layer of the body, they are most susceptible to pathogenic invasion. If the skin and hair are affected, the lungs lose their dispersing and descending function, leading to impaired lung Qi and a disruption of the lung's wei qi function, resulting in a common cold. The nose is the orifice of the lungs, and symptoms are characterized by nasal congestion, runny nose, aversion to wind, fever, and headache, occurring in all seasons. This disease is often diagnosed as wind-heat or wind-cold colds, with wind-heat colds being more common. If it is widespread, it is called "seasonal influenza."

[0010] The classic Chinese medicine text, *Huangdi Neijing*, contains numerous descriptions of symptoms similar to the common cold. For example, in *Suwen* (Plain Questions), "On the Circulation of Vital Energy," it states, "Due to exposure to wind and dew, chills and fever arise." The Song Dynasty scholar Zhao Ji, in his *Shengji Zonglu* (General Records of Sacred Relief), states, "Wind-heat is caused by wind-evil and heat. It lodges in the skin, hair, and blood vessels, and then enters the lung meridian. It causes a burning sensation in the head and face, skin pain, cough, dry throat, and dysfunction of the upper burner; hence it is called wind-heat." This further explains the pathogenesis of the common cold, stating that the pathogenic factor can invade the lung meridian, leading to dysfunction of the upper burner. The Qing Dynasty scholar Ye Tianshi, in his *External Febrile Diseases*, states, "Before the pathogenic factor has spread to the pericardium, it is still in the lungs. The lungs govern qi, and it is associated with the skin and hair; therefore, it is said to be on the exterior. When it is on the exterior, mild, pungent, and cooling remedies are initially used." The *Ling Shu* (Spiritual Pivot), in its chapter "The Origin of All Diseases," states: "Wind, rain, cold, and heat cannot harm a person without a deficiency; evil cannot harm a person alone. Those who suddenly encounter strong winds and torrential rains but do not fall ill are because they are not deficient; therefore, evil cannot harm a person alone. This must be due to the wind of deficiency and the body's own weakness, where both are combined, thus allowing the evil to invade the body." This indicates that the six pathogenic factors or seasonal viruses can invade the body and cause colds, and apart from the particularly strong pathogenic factors, it is always related to the imbalance of the body's vital energy. At the initial stage of infection, patients often exhibit underlying pathological characteristics such as weakened lung and defensive qi.

[0011] Based on the above TCM theory, "when pathogenic heat invades the body, it first attacks the lungs." Weakness in the lung's defensive qi is one of the internal causes of viral infections, including COVID-19. Therefore, strengthening the defensive qi and eliminating pathogenic factors are important aspects of preventing and treating this disease. The location of the common cold is in the lung's defensive qi; its basic pathogenesis is the invasion of external pathogens and dysfunction of the lung's defensive qi, leading to disharmony of the defensive qi and impaired lung function, resulting in cough. Therefore, clearing the lungs and relieving cough is another important aspect of treating this disease. As the first aspect of this invention, the TCM composition of this invention is based on the above analysis of the pathogenesis of viral respiratory infections and COVID-19. It is formulated according to the etiological characteristics of various diseases and based on syndrome differentiation and treatment. Furthermore, through network pharmacology, a "drug-component-intersecting target" relationship is constructed, verifying the mechanism of action of the TCM composition with the common cold virus and the novel coronavirus, forming a TCM composition with multiple effects.

[0012] On the one hand, in the traditional Chinese medicine composition of this invention, honeysuckle, forsythia, mulberry leaf, and chrysanthemum are the principal herbs, possessing both the effects of clearing heat and dispelling pathogens with their pungent and cooling properties, as well as the functions of dispelling foulness and detoxifying with their aromatic properties. Mulberry leaf is good at traversing the lung channels, clearing lung heat and stopping cough. Raw ephedra, roasted bitter apricot kernel, and platycodon root clear lung qi and stop cough, while bupleurum root is used for exogenous fever, exerting the function of relieving exterior symptoms and reducing fever. These four herbs together serve as assistant herbs. Reed rhizome clears heat, generates fluids, and quenches thirst; honey loquat leaf clears the lungs and stops cough; verbena clears heat and detoxifies; and codonopsis root strengthens the spleen and aids digestion to assist the qi of the spleen and lungs, supporting the body's resistance and eliminating pathogens. These herbs together serve as adjuvant herbs. Raw licorice root, unprocessed, is used for its heat-clearing and detoxifying properties, working synergistically with raw ephedra, roasted bitter apricot kernel, and platycodon root to soothe the throat and stop cough, harmonizing the other herbs. These herbs together achieve the functions of dispelling wind and clearing heat, clearing the lungs and stopping cough, and supporting the body's resistance and eliminating pathogens.

[0013] On the other hand, the specific effective components of each medicinal herb in the above-mentioned traditional Chinese medicine composition are as follows:

[0014] Mulberry leaves contain chemical components including flavonoids, organic acids and similar compounds, polyphenols, polysaccharides, alkaloids, volatile oils, amino acids and vitamins. They have antipyretic, analgesic, anti-inflammatory, hypoglycemic, anti-atherosclerotic, tumor-inhibiting and anti-aging effects.

[0015] Chrysanthemum contains chemical components including flavonoids, volatile oils, organic acids, anthraquinones, phenylpropanoids, triterpenes and steroids, sugars and glycosides. It has effects on the cardiovascular system, protecting the nerves and liver, antioxidation, regulating the body's immunity, antibacterial, antiviral, anti-inflammatory, antitumor and hypoglycemic effects.

[0016] Forsythia contains chemical components including phenylethyl glycosides, lignins, terpenes and volatile oils, flavonoids, phenolic acids and their derivatives, sterols and alkaloids, and has antibacterial, antiviral, antipyretic and analgesic, antioxidant, hepatoprotective and immunomodulatory effects.

[0017] Honeysuckle contains organic acids, flavonoids, iridoids, triterpenoid saponins, and volatile oils. It has anti-inflammatory, lipid-lowering, blood sugar-lowering, antioxidant, antiviral, hepatoprotective, choleretic, and immune-enhancing effects.

[0018] Platycodon grandiflorus contains chemical components including saponins, volatile oils, flavonoids, fatty acids, amino acids, vitamins, polysaccharides, and inorganic elements. It has anti-inflammatory, antitumor, pancreatic exocrine gland secretion-promoting, lipid-lowering, antitussive, expectorant, and antioxidant effects.

[0019] Bitter almonds contain chemical components including cyanides, fats, volatile oils, amino acids, and vitamins. They have antitussive, antiasthmatic, anti-inflammatory, analgesic, antitumor, antioxidant, and anti-organ fibrosis effects, as well as effects on the immune system and cardiovascular diseases.

[0020] Loquat leaves contain chemical components including flavonoids, triterpenoids, organic acids, volatile oils, and inorganic elements. They have anti-inflammatory, expectorant, antitussive, anti-pulmonary fibrosis, antioxidant, liver-protective, and antiemetic effects.

[0021] Raw ephedra contains chemical components including alkaloids, flavonoids, volatile oils, organic acids, polysaccharides, and tannins. It has antitussive, antiasthmatic, diaphoretic, diuretic, anti-allergic, antioxidant, and blood pressure and smooth muscle effects.

[0022] Bupleurum chinense contains chemical components including saponins, volatile oils, flavonoids, polysaccharides, and coumarins, and has antidepressant, anti-inflammatory, hepatoprotective, antitumor, and cardioprotective and renal-protective effects.

[0023] Verbena officinalis contains chemical components including flavonoids, iridoids, triterpenoids and diterpenoids, phenylethanol glycosides, sterols and volatile chemical components, and has antibacterial, antiviral, immunomodulatory, antitumor and neuroprotective effects.

[0024] Reed rhizome contains polysaccharides, steroids, anthraquinones, flavonoids, volatile components, small molecules, and alkaloids. It has hepatoprotective, antioxidant, antitumor, lipid metabolism-improving, and liver and kidney-protective effects.

[0025] Codonopsis pilosula, a herb containing cyclic peptides, sugars, trace elements, glycosides, amino acids, phospholipids, fatty acids, oils, volatile oils, and sterols, has immunomodulatory, hypoglycemic, myocardial protective, anti-stress, anti-inflammatory, anti-tumor, and antitussive effects.

[0026] Raw licorice contains chemical components including triterpenoid saponins, flavonoids, coumarins, and polysaccharides. It has antioxidant, antitumor, antibacterial, neuroprotective, anti-inflammatory, antidepressant, cardiovascular protective, and immunomodulatory effects.

[0027] The analysis and research on the mechanism of action of the above-mentioned active ingredients and viral targets show that the synergistic effect of each active ingredient can effectively target the common cold virus and the novel coronavirus, achieving a definite therapeutic effect.

[0028] In summary, the traditional Chinese medicine composition of the present invention has a precise target mechanism, a precise mechanism of action, and a precise therapeutic effect, both macroscopically and microscopically.

[0029] As a second aspect of the present invention, the pharmaceutical composition of the present invention comprises an extract of the above-mentioned traditional Chinese medicine composition and a pharmaceutically acceptable carrier; its formulation includes granules, capsules or tablets.

[0030] As a third aspect of the present invention, the method for preparing the above-mentioned pharmaceutical composition includes the following steps:

[0031] (1) Combined extraction: Take mulberry leaves, chrysanthemum, forsythia, honeysuckle, platycodon, bitter almond, honey loquat leaves, ephedra, bupleurum, verbena, reed rhizome, codonopsis and licorice by weight, combine them and extract with water, filter the obtained extract, concentrate and centrifuge to obtain the centrifuged solution;

[0032] (2) Prepare a pharmaceutical composition by adding pharmaceutically acceptable excipients.

[0033] Furthermore, the specific operation of step (1) is as follows:

[0034] Take mulberry leaves, chrysanthemum, forsythia, honeysuckle, platycodon, bitter almond, honey-processed loquat leaves, raw ephedra, bupleurum, verbena, reed rhizome, codonopsis, and raw licorice root by weight. Add 6 times the amount of water to the total herbal composition and decoct to extract. Soak for 30 minutes, decoct for 20 minutes, and filter to obtain filtrate one and residue. Add 5 times the amount of water to the residue again, decoct for 20 minutes, and filter to obtain filtrate two. Combine filtrate one and filtrate two, concentrate under reduced pressure, concentrate at 60℃ to a relative density of 1.20, and then centrifuge using a straight tube high-speed centrifuge to obtain the centrifuged solution.

[0035] Furthermore, when the pharmaceutical composition prepared in step (2) is in the form of granules, dextrin is taken and placed in a fluidized bed. When the material temperature rises, the centrifuged solution obtained in step (1) is started to be spray-dried (e.g., the liquid feed rate is 80-150 r / min) to obtain granules.

[0036] The specific steps are as follows:

[0037] Dextrin was placed in a fluidized bed, and the inlet air temperature was controlled at 100℃. When the dextrin temperature reached 70℃, the solution was centrifuged and spray-dried. The addition rate of the extract was controlled at 80-150 r / min and the atomization pressure was 0.2 MPa externally and 0.15 MPa internally. After the spray drying was completed, the mixture was dried at 70℃ for 1 hour to produce 1000 g of the pharmaceutical composition granules.

[0038] As a fourth aspect of the present invention, the quality detection method of the above-mentioned pharmaceutical composition includes thin-layer chromatography detection and high-performance liquid chromatography detection.

[0039] On the one hand, the above-mentioned thin-layer chromatography detection includes the detection of ephedra, licorice, forsythia, and codonopsis. Among them, the thin-layer chromatography developing solvent for ephedra is dichloromethane-methanol-concentrated ammonia solution with a volume ratio of 20:5:0.5; the thin-layer chromatography developing solvent for licorice is petroleum ether-toluene-ethyl acetate-glacial acetic acid solution with a volume ratio of 10:20:7:0.5; the thin-layer chromatography developing solvent for forsythia is chloroform-methanol solution with a volume ratio of 49:1; and the thin-layer chromatography developing solvent for codonopsis is n-butanol-glacial acetic acid-water solution with a volume ratio of 4:1:1.

[0040] Specifically, the thin-layer chromatography detection procedure for ephedra is as follows:

[0041] (1) Preparation of test solution: Dissolve 10g of drug composition in water, adjust pH to 11-12 with sodium hydroxide test solution, extract twice with dichloromethane, 20mL each time, combine the dichloromethane extracts, evaporate to dryness, add 2mL of methanol to dissolve the residue to obtain the test solution.

[0042] (2) Preparation of negative test sample: Prepare a negative test sample without ephedrine drug composition according to step (1);

[0043] (3) Preparation of reference solution: Take Ephedra reference powder, add 50 mL of water and reflux for 1 hour, cool, and filter to obtain Ephedra reference solution.

[0044] (4) Thin-layer chromatography test: Take 10 μL of each of the above three solutions and spot them on the same silica gel G thin-layer plate. Use dichloromethane-methanol-concentrated ammonia solution (volume ratio of 20:5:0.5) as the developing solvent, develop, remove, air dry, spray with 2% ninhydrin solution, and heat at 105℃ until the spots are clearly visible.

[0045] The specific procedures for the thin-layer chromatography detection of the above-mentioned licorice are as follows:

[0046] (1) Preparation of test solution: Dissolve 10g of drug composition in 2mL hydrochloric acid, heat in water bath at 100℃ for 1 hour, remove, cool, filter, extract the filtrate with water-saturated n-butanol 3 times, 30mL each time, combine the n-butanol extracts, evaporate to dryness in water bath, add 2ml methanol to dissolve the residue to obtain the test solution.

[0047] (2) Preparation of negative test sample: Prepare a negative test sample without licorice drug composition according to step (1);

[0048] (3) Preparation of reference solution: Take licorice reference powder, add 100 mL of water and reflux for 30 minutes, filter, add 3 mL of hydrochloric acid to the filtrate, heat in a water bath at 100℃ for 1 hour, take it out, cool, filter, extract the filtrate with water-saturated n-butanol 3 times, 30 mL each time, combine the n-butanol extracts, evaporate to dryness, add 2 mL of methanol to dissolve the residue, and use it as licorice reference solution;

[0049] (4) Thin-layer chromatography test: Take 10 μL of each of the above three solutions and spot them on the same silica gel G thin-layer plate. Use petroleum ether (60-90℃)-toluene-ethyl acetate-glacial acetic acid (volume ratio of 10:20:7:0.5) as the developing solvent, develop, remove, air dry, spray with 10% sulfuric acid ethanol solution, and heat at 105℃ until the spots are clearly visible.

[0050] The specific procedures for the thin-layer chromatography detection of Forsythia suspensa are as follows:

[0051] (1) Preparation of test solution: Dissolve the drug composition in 30 mL of methanol, sonicate for 30 min, filter, evaporate the filtrate to dryness, dissolve the residue in 1 mL of methanol to obtain the test solution;

[0052] (2) Preparation of negative test solution: Take the drug composition that does not contain Forsythia and prepare the negative test solution according to the test solution preparation method;

[0053] (3) Preparation of reference medicinal material solution: Take 3g of Forsythia suspensa reference medicinal material powder, add 10mL of methanol, and prepare the reference medicinal material solution according to the preparation method of the test sample solution;

[0054] (4) Thin-layer chromatography test: Take 20 μL of the above three solutions and spot them on the same silica gel G thin-layer plate. Use chloroform-methanol with a volume ratio of 49:1 as the developing solvent, develop, remove, air dry, spray with 10% sulfuric acid ethanol solution, bake at 105℃ for 5 minutes, place in ultraviolet 366nm, and develop color.

[0055] The specific procedures for the thin-layer chromatography detection of the above-mentioned Codonopsis pilosula are as follows:

[0056] (1) Preparation of test solution: Dissolve the drug composition in 30 mL of methanol, wet soak, sonicate for 30 min, filter, evaporate to dryness, dissolve the residue in 1 mL of methanol to obtain the test solution;

[0057] (2) Preparation of negative test solution: Take the drug composition that does not contain Codonopsis pilosula and prepare a negative test solution according to the test solution preparation method;

[0058] (3) Preparation of reference medicinal material solution: Take 3g of reference medicinal material powder, add 10mL of methanol, and prepare the reference medicinal material solution according to the preparation method of the test sample solution;

[0059] (4) Thin-layer chromatography test: Take 5 μL of the above three solutions and spot them on the same silica gel G thin-layer plate. Use n-butanol-glacial acetic acid-water as the developing solvent with a volume ratio of 4:1:1, pre-saturate for 15 minutes, develop, remove, air dry, spray with 0.2% ninhydrin solution, and heat at 105℃ until the spots are clearly visible.

[0060] On the other hand, the above-mentioned high-performance liquid chromatography detection method includes the following steps:

[0061] (1) Preparation of reference solution: Take ephedrine hydrochloride standard and pseudoephedrine hydrochloride standard and prepare reference solution;

[0062] (2) Preparation of test solution: Dissolve the drug composition in water, add sodium hydroxide solution and sodium chloride, distill, collect the distillate, add hydrochloric acid solution beforehand, collect, add water to make up to volume, shake well, filter, and take the filtrate to obtain the test solution; take the drug composition that does not contain ephedrine and prepare the negative test solution according to the test solution preparation method;

[0063] (3) Plotting the standard curve: Take the reference standard, mix and dilute it to prepare reference standard solutions of different concentrations, and plot the standard curve with concentration as the abscissa x and peak area as the ordinate y. The regression equation of ephedrine hydrochloride is y = 9353.6x - 4.2432, and the regression equation of pseudoephedrine hydrochloride is y = 10876x - 29.694.

[0064] (4) Detection: Take the reference solution from step (1) and the test solution from step (2) and inject them into the high performance liquid chromatograph. The chromatographic conditions are phenyl column, column temperature: 40℃, detection wavelength: 210nm, elution with acetonitrile as mobile phase A and 0.2% phosphoric acid aqueous solution as mobile phase B. The elution program is as follows: 0-30min, A:B is maintained at 3:97; 30-35min, A:B is converted to 95:5; 35-45min, A:B is maintained at 95:5; 45-50min, A:B is converted to 3:97.

[0065] (5) Calculate the content: Based on the standard curve equation in step (3), calculate the content of ephedrine and pseudoephedrine in the test solution.

[0066] Furthermore, in the above-mentioned high-performance liquid chromatography (HPLC) detection method, the detection linear range of ephedrine hydrochloride is 0.0544–15.7286 mg / mL, and the detection linear range of pseudoephedrine hydrochloride is 0.0798–11.945 mg / mL.

[0067] The specific operation of step (1) above is as follows:

[0068] (1) Preparation of reference solution: Accurately weigh 0.0308 g of 100% pure ephedrine hydrochloride standard and 0.0238 g of 99.8% pure pseudoephedrine hydrochloride standard, and dissolve them in anhydrous methanol to prepare a reference solution containing 3.08 mg / mL ephedrine hydrochloride and 2.38 mg / mL pseudoephedrine hydrochloride;

[0069] The specific steps for step (2) are as follows:

[0070] (2) Preparation of test solution: Dissolve 10g of drug composition in 100mL of water, add 120mL of 20% sodium hydroxide solution and 7.5g of sodium chloride, distill, collect the distillate, add 5mL of 0.5mol / L hydrochloric acid solution beforehand, collect 120mL, add water to make up to volume, shake well, filter through a 0.45μm microporous membrane, and take the filtrate to obtain the test solution; take the drug composition that does not contain ephedrine and prepare the negative test solution according to the test solution preparation method;

[0071] Specifically, in step (4) above, the chromatographic conditions of the high performance liquid chromatograph are as follows: column temperature: 40℃, detection wavelength: 210nm, injection volume: 10μL, theoretical plate number of ephedrine hydrochloride is higher than 3000, the phenyl column is an Agilent Zorbax SB-Phenyl column (250nm×4.6mm, 5μm), and the flow rate is 1mL / min.

[0072] As the fifth aspect of this invention, the above-mentioned traditional Chinese medicine composition and its pharmaceutical composition can be prepared as a drug for treating mild COVID-19 and epidemic viral colds, especially suitable for people with wind-heat attacking the lungs, wind-cold transforming into heat, lung qi deficiency or insufficient vital energy. The main symptoms are sore throat, cough, fever, nasal congestion, runny nose, fatigue, etc. caused by exogenous wind evil, with red tongue, thin yellow coating and floating pulse.

[0073] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages:

[0074] (1) The Chinese medicine composition targets multiple pathogenesis, differentiates syndromes and treats accordingly, and has a reasonable prescription. It has the effects of dispelling wind and clearing heat, ventilating the lungs and relieving cough, and supporting the body's resistance and eliminating pathogens. It is used for mild COVID-19 and influenza. Clinical trials have shown that the medication is safe and has no obvious adverse reactions.

[0075] (2) A connection between “drug-component-intersection target” was established, clarifying the mechanism of action of traditional Chinese medicine composition on influenza and COVID-19, with definite curative effect;

[0076] (3) The drug composition formulation is flexible, allowing for personalized medication, and the preparation method is simple, which is conducive to large-scale preparation;

[0077] (4) By combining thin-layer chromatography with high-performance liquid chromatography, a comprehensive quality control method for both qualitative and quantitative analysis was established. The detection method is precise and simple to operate and has the advantages of strong specificity, high sensitivity, high accuracy, wide durability, high precision, good stability and high identification efficiency. It is beneficial to control the quality of drug compositions and ensure their safety and stability. Attached Figure Description

[0078] Figure 1 This is an intersection diagram of the effective ingredient targets of the Qingxuan Jiedu Granules of this invention and COVID-19-related targets;

[0079] Figure 2 This is a drug-component-target network diagram;

[0080] Figure 3 PPI network diagram;

[0081] Figure 4 GO analysis diagram;

[0082] Figure 5 A graph showing KEGG pathway enrichment analysis;

[0083] Figure 6 Docking diagram of EGFR-β-sitosterol molecules;

[0084] Figure 7 Docking diagram of IL6-β-sitosterol molecules;

[0085] Figure 8 This is a docking diagram of TNF-β-sitosterol molecules;

[0086] Figure 9 Diagram of TNF-kaempferol molecular docking;

[0087] Figure 10 The images show thin-layer chromatograms of raw ephedra, where a: thin-layer chromatogram of raw ephedra, b: thin-layer chromatogram at 10℃, c: thin-layer chromatogram at 22℃, d: thin-layer chromatogram at 30% humidity, e: thin-layer chromatogram at 60% humidity, f: G-plate manufacturer is Qingdao Ocean, g: G-plate manufacturer is Yantai Huanghai;

[0088] Figure 11The images show thin-layer chromatograms of raw licorice, where a: thin-layer chromatogram of raw licorice, b: thin-layer chromatogram at 10℃, c: thin-layer chromatogram at 25℃, d: thin-layer chromatogram at 30% humidity, e: thin-layer chromatogram at 60% humidity, f: G-plate manufacturer is Qingdao Ocean, g: G-plate manufacturer is Yantai Huanghai;

[0089] Figure 12 The images show thin-layer chromatograms of Forsythia suspensa, where a: Forsythia suspensa thin-layer chromatogram, b: thin-layer chromatogram at 10℃, c: thin-layer chromatogram at 23℃, d: thin-layer chromatogram at 30% humidity, e: thin-layer chromatogram at 60% humidity, f: G board manufacturer is Qingdao Ocean, g: G board manufacturer is Yantai Huanghai.

[0090] Figure 13 The images show thin-layer chromatograms of Codonopsis pilosula, where a: Codonopsis pilosula thin-layer chromatogram, b: thin-layer chromatogram at 10℃, c: thin-layer chromatogram at 23℃, d: thin-layer chromatogram at 30% humidity, e: thin-layer chromatogram at 60% humidity, f: G plate manufacturer is Qingdao Ocean, g: G plate manufacturer is Yantai Huanghai;

[0091] Figure 14 This is the high-performance liquid chromatogram of the test solution. Detailed Implementation

[0092] The present invention will be further described below with reference to specific embodiments.

[0093] Example 1: Preparation of Qingxuan Jiedu Granules

[0094] 1. The raw materials and standards of the medicinal materials used in the traditional Chinese medicine composition of this invention.

[0095] Mulberry leaves (Sangye MORI FOLIUM) are the dried leaves of the mulberry tree (Morus alba L.), a plant in the Moraceae family. They are harvested after the first frost, impurities are removed, and they are sun-dried. They comply with all relevant provisions of the Pharmacopoeia of the People's Republic of China, Volume 1, page 310, under the section on Mulberry Leaves - Prepared Slices.

[0096] Chrysanthemum (Juhua CHRYSANTHEMI FLOS) is the dried capitulum of Chrysanthemum morifolium Ramat., a plant in the Asteraceae family. It is harvested in batches from September to November when the flowers are in full bloom, and then dried in the shade, baked, or steamed and then sun-dried. It conforms to all relevant provisions under the Chrysanthemum - Prepared Slices section on page 323 of Part I of the 2020 edition of the Pharmacopoeia of the People's Republic of China.

[0097] Forsythia fructus (Lianqiao FORSYTHIAE FRUCTUS) is the dried fruit of Forsythia suspensa (Thunb.) Vahl, a plant in the Oleaceae family. It conforms to all relevant provisions of the 2020 edition of the Pharmacopoeia of the People's Republic of China, Volume I, page 177, under the section on Forsythia fructus processed products.

[0098] Honeysuckle (Jinyinhua LONICERAE JAPONICAE FLOS) is the dried flower bud or newly opened flower of *Lonicera japonica* Thunb., a plant in the Caprifoliaceae family. It is harvested in early summer before the flowers open and then dried. It conforms to all relevant regulations under the Honeysuckle - Prepared Slices section on page 230 of Part I of the 2020 edition of the *Pharmacopoeia of the People's Republic of China*.

[0099] Platycodon grandiflorum (Jacq.) A.DC., a plant in the Campanulaceae family, is the dried root of the plant. It is harvested in spring and autumn, washed, and the fibrous roots are removed. While still fresh, the outer skin is peeled off, or left on, and then dried. It conforms to the relevant provisions of the Platycodon grandiflorum-processed medicinal materials section on page 289 of Part I of the 2020 edition of the Pharmacopoeia of the People's Republic of China.

[0100] Bitter almonds (Kuxingren ARMENIACAE SEMEN AMARUM) are the dried, mature seeds of *Prunus armeniaca* L. var. *ansu* Maxim., a plant in the Rosaceae family. Mature fruits are harvested in summer, the pulp and shell are removed, and the seeds are extracted and dried. They comply with all relevant regulations for bitter almonds (processed medicinal materials) on page 210 of Part I of the 2020 edition of the *Pharmacopoeia of the People's Republic of China*.

[0101] Reed rhizome (Lugen PHRAGMITIS RHIZOMA) is the fresh or dried rhizome of the grass Phragmites communis Trin. It can be harvested year-round. Remove the buds, fibrous roots, and membranous leaves, and use it fresh or dried. It conforms to the relevant provisions of the Reed Rhizome - Prepared Slices section on page 171 of Part I of the 2020 edition of the Pharmacopoeia of the People's Republic of China.

[0102] Honey loquat leaves (Pipaye ERIOBOTRYAE FOLIUM) are the dried leaves of Eriobotrya japonica (Thunb.) Lindl., a plant in the Rosaceae family. They can be harvested year-round. After being sun-dried to about 70-80% dryness, they are bundled into small bunches and sun-dried further. They comply with all relevant regulations for honey loquat leaves under the Loquat Leaf - Prepared Slices section on page 213 of Part I of the 2020 edition of the Pharmacopoeia of the People's Republic of China.

[0103] Raw ephedra (Mahuang EPHEDRAE HERBA) is the dried herbaceous stem of *Ephedra equisetina* Bge., a plant in the Ephedraceae family. The green herbaceous stems are harvested in autumn and sun-dried. It conforms to all relevant provisions of the *Pharmacopoeia of the People's Republic of China*, 2020 edition, Part I, page 333, under the section on Ephedra-processed Herbs.

[0104] Chaihu BUPLEURI RADIX is the dried root of Bupleurum chinense DC., a plant belonging to the Apiaceae family. It is harvested in spring and autumn, with stems, leaves, and soil removed before drying. It conforms to all relevant provisions under the "Chaihu BUPLEURI RADIX - Processed Roots" section on page 293 of Part I of the 2020 edition of the Pharmacopoeia of the People's Republic of China.

[0105] Verbena officinalis (Mabiancao VERBENAE HERBA) is the dried aerial part of the plant Verbena officinalis L., belonging to the Verbenaceae family. It is harvested during the flowering period from June to August, impurities are removed, and it is sun-dried. It conforms to the relevant provisions of the Verbena officinalis-processed herbs section on page 53 of Part I of the 2020 edition of the Pharmacopoeia of the People's Republic of China.

[0106] Taizishen (Pseudostellaria heterophylla (Miq.) Paxex Paxet Hoffm.), a plant in the Caryophyllaceae family, is the dried tuberous root. It is harvested in summer when most of the stems and leaves have withered. The roots are washed, the fibrous roots are removed, and the roots are briefly blanched in boiling water before being sun-dried or directly sun-dried. It conforms to the relevant provisions of the 2020 edition of the Pharmacopoeia of the People's Republic of China, Volume I, page 69, under the section on Taizishen - Processed Herbs.

[0107] Raw licorice (Gancao RADIX ET RHIZOMA GLYCYRRHIZAE) is the dried root and rhizome of Glycyrrhiza uralensis Fisch., a plant belonging to the Fabaceae family. It is harvested in spring and autumn, the fibrous roots are removed, and it is sun-dried. It conforms to all relevant provisions of the 2020 edition of the Pharmacopoeia of the People's Republic of China, Volume I, page 88, under the section on licorice tablets and processed medicinal materials.

[0108] The herbal composition for clearing heat and detoxifying contains, by weight, 10 parts mulberry leaf, 10 parts chrysanthemum, 10 parts forsythia, 10 parts honeysuckle, 6 parts platycodon, 10 parts bitter almond, 10 parts honey loquat leaf, 3 parts raw ephedra, 8 parts bupleurum, 12 parts verbena, 15 parts reed rhizome, 10 parts codonopsis, and 6 parts raw licorice.

[0109] 2. Particle preparation

[0110] The preparation method of the pharmaceutical composition granules is as follows:

[0111] Weigh all raw materials according to their mass proportions, mix them, add 6 times the amount of water to the total herbal composition, soak for 30 minutes, decoct for 20 minutes, filter, and obtain filtrate one and residue; add 5 times the amount of water to the residue again, decoct for 20 minutes, filter, and obtain filtrate two; combine filtrate one and filtrate two, concentrate under reduced pressure, concentrate at 60℃ to a relative density of 1.20, and then centrifuge using a straight tube high-speed centrifuge to obtain the centrifuged solution; place dextrin in a fluidized bed, control the inlet air temperature at 100℃, and when the dextrin temperature rises to 70℃, start feeding the centrifuged solution for spray drying, control the extraction rate of the extract at 80-150 r / min and the atomization pressure at 0.2 MPa externally and 0.15 MPa internally, and after the spray drying is completed, continue drying at 70℃ for 1 hour to produce a total of 1000g, which is the Qingxuan Jiedu Granules.

[0112] Example 2: Network pharmacology study of Qingxuan Jiedu granules

[0113] 1. Materials and Methods

[0114] 1.1 Collection of active ingredients of Qingxuan Jiedu Granules and their COVID-19-related targets

[0115] The effective components of each herb in the Qingxuan Jiedu Granules prescription (containing mulberry leaf, chrysanthemum, forsythia, honeysuckle, platycodon, bitter almond, loquat leaf, raw ephedra, bupleurum, verbena, reed rhizome, codonopsis, and raw licorice) were retrieved from the TCMSP database (https: / / old.tcmsp-e.com / tcmsp.php). Screening criteria were set as follows: oral bioavailability (OB) ≥ 30% and drug-likeness (DL) ≥ 0.18. Literature was also reviewed to supplement the list with compounds that did not meet the screening criteria but were major active ingredients of the aforementioned herbal medicines or had reported anti-COVID-19, antiviral, or anti-inflammatory effects, and these were considered as effective active ingredients. Furthermore, the TCMSP database was used to collect targets for these effective active ingredients. In this study, components without targets in the TCMSP were imported into the Pubchem database (https: / / pubchem.ncbi.nlm.nih.gov / ) to retrieve their SMILES numbers. These SMILES numbers were then imported into the SwissTargetPrediction database (http: / / swisstargetprediction.ch / ), with "homo sapiens" selected as the research species. Targets with a probability greater than 0 were chosen as the effective active ingredient targets. Disease-related targets were searched in the GeneCards database (https: / / www.genecards.org / ) using "COVID-19" as the keyword. Online analysis of the intersection between the effective ingredient targets of Qingxuan Jiedu Granules and COVID-19-related targets was conducted using the Venny website (https: / / bioinfogp.cnb.csic.es / tools / venny / ) to identify potential targets for treating COVID-19 with Qingxuan Jiedu Granules.

[0116] 1.2 Construction of Drug-Component-Intersection Target Network

[0117] The Chinese herbal medicines and components of Qingxuan Jiedu Granules, as well as the intersection target data, were imported into Cytoscape 3.6.1 software to construct a "component-intersection target" network.

[0118] 1.3 Construction of PPI Network for Intersecting Targets

[0119] Import the intersection target points into the string database (https: / / cn.string-db.org / ), limit the species to "human", set the confidence level to high confidence (0.700) as the indicator, and leave other parameters at default to construct a PPI network. Import the results into Cytoscape 3.6.1 software and use its Network Analyzer function for visualization analysis.

[0120] 1.4 GO (Gene Ontology) analysis and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway enrichment analysis

[0121] Intersecting targets were imported into the Metascape (http: / / metascape.org / ) database for GeneOntology (GO) analysis and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway enrichment analysis. The species was limited to "Homo sapiens," and a p-value of <0.01 was set to obtain the results of biological process (BP), molecular function (MF), cellular component (CC), and signaling pathway enrichment analysis (KEGG). The results were then visualized using the Bioinformatics (http: / / www.bioinformatics.com.cn / ) website.

[0122] 1.5 Component-Target Molecular Docking

[0123] Based on the constructed network and literature review, the core proteins TNF, EGFR, IL6, AR, and STATE3, as well as key components quercetin, luteolin, kaempferol, beta-sitosterol, and troxerutin were screened. The core proteins were retrieved from the PDB database (https: / / www.rcsb.org / ), and their protein structure PDB files were downloaded. The key components of Qingxuan Jiedu Granules were retrieved from the TCMSP database (https: / / old.tcmsp-e.com / tcmsp.php), and their 3D structure mol2 files were downloaded. In PyMOL software, the core proteins TNF, EGFR, IL6, AR, and STATE3 underwent dehydration and hydrogenation operations. Then, the key components of Qingxuan Jiedu Granules and the core proteins were preprocessed using the molecular docking software AutoDock, including charge calculation and conformation recognition, and the format was converted to pdbqt format before docking. The docking results were then analyzed, and binding energies <0 kJ·mol⁻¹ were selected. -1 Furthermore, the resulting complexes exhibit a high number of hydrogen bonds. Finally, the molecular docking results were visualized using PyMOL software.

[0124] 2. Results

[0125] 2.1 Collection of effective active ingredients and COVID-19-related targets

[0126] Based on the TCMSP database and literature, a total of 232 effective active ingredients of Qingxuan Jiedu Granules were retrieved, corresponding to 621 target sites. Based on the GeneCards database, a total of 4600 targets related to COVID-19 were retrieved, and online analysis using the Venny website yielded 210 overlapping targets, such as... Figure 1 .

[0127] 2.2 Construction of Drug-Component-Intersection Target Network

[0128] A network diagram of the "drug-component-intersecting target" relationship was constructed using Cytoscape 3.6.1 software. (See...) Figure 2A total of 167 components are associated with the intersection target points. The yellow nodes in the network represent the 13 traditional Chinese medicines in Qingxuan Jiedu Granules. The diamond-shaped nodes represent the 167 effective active ingredients in Qingxuan Jiedu Granules that are associated with the intersection target points. The blue diamond-shaped nodes surrounding each yellow node (medicine) represent the unique components of each traditional Chinese medicine, while the red diamond-shaped nodes represent components shared by two or more traditional Chinese medicines. The blue circular nodes represent the intersection target points. The lines connecting two nodes indicate their interrelationships, and the size of the node indicates its correlation with other nodes; a larger node indicates more connections to other nodes. The higher the degree value of a node in the network relationship, the greater its influence within the network. The top ten active ingredients with the highest degree values ​​are quercetin, luteolin, kaempferol, beta-sitosterol, troxerutin, (-)-epigallocatechin-3-gallate, naringenin, isorhamnetin, vesitol, and stigmasterol, suggesting these components may be key ingredients in Qingxuan Jiedu Granules' treatment of COVID-19. Within its network system, the top ten intersection target points with the highest degree values ​​are AR, PRKACA, F10, GSK3B, DPP4, F2, ACHE, NOS3, CASP3, and RELA, suggesting these targets may be potential targets for the therapeutic effect of Qingxuan Jiedu Granules on COVID-19.

[0129] 2.3 Construction of PPI network for intersection target points

[0130] The PPI network of intersection target points constructed using the string database and Cytoscape 3.6.1 software is as follows: Figure 3 As shown in the PPI network diagram, after removing free nodes, there are 196 nodes, each representing a potential target. The connections between nodes represent protein-protein interactions. The node size and color are adjusted with the degree value; the higher the degree value, the larger the node and the warmer the color. The connections between nodes are adjusted with the strength of the interaction between the two nodes; the stronger the interaction, the thicker the connection and the cooler the color. The top ten targets with the highest degree values ​​are STAT3, SRC, IL6, HSP90AA1, TP53, TNF, EGFR, AKT1, MAPK3, and CTNNB1, indicating that these targets may be potential targets for the therapeutic effect of Qingxuan Jiedu Granules on COVID-19.

[0131] 2.4 GO (Gene Ontology) analysis and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway enrichment analysis

[0132] GO functional enrichment analysis yielded 2220 biological process (BP) entries, 130 cellular composition (CC) entries, and 207 molecular function (MF) entries. Biological process analysis showed that the targets of Qingxuan Jiedu granules in treating COVID-19 are mainly related to cellular responses to nitrogen compounds, positive regulation of inflammatory cell migration, cellular responses to cytokine stimulation, responses to bacterial-derived molecules, positive regulation of responses to external stimuli, positive regulation of protein phosphorylation, cell activation, protein phosphorylation, and cellular responses to chemical stress. Cellular composition analysis indicated that Qingxuan Jiedu granules may act on membrane rafts, receptor complexes, lateral membranes, perinuclear regions of the cytoplasm, vesicles, cell apex, lumen, endocytic vesicles, transcriptional regulatory complexes, and the cell body in treating COVID-19. Molecular functional analysis suggests that the therapeutic effects of Qingxuan Jiedu granules on COVID-19 may be primarily related to protein serine / threonine / tyrosine kinase activity, cytokine receptor binding, kinase binding, protein tyrosine kinase activity, protein domain-specific binding, cell adhesion molecule binding, RNA polymerase II-specific DNA binding, transcription factor binding, scaffold protein binding, phosphatase binding, and sulfur compound binding. (See [link to relevant documentation]). Figure 4 .

[0133] KEGG pathway enrichment analysis yielded 207 pathways, which were then sorted by their Log(q-value) values ​​from smallest to largest (smaller Log(q-value) values ​​indicate greater correlation). The top 20 pathways were selected. See [link to relevant documentation]. Figure 5 Enrichment studies revealed that the mechanism by which Qingxuan Jiedu Granules treat COVID-19 is mainly related to cancer pathways, lipids and atherosclerosis, Kaposi's sarcoma-associated herpesvirus infection, hepatitis C, human cytomegalovirus infection, prostate cancer, the AGE-RAGE signaling pathway in diabetic complications, hepatitis B, proteoglycans in cancer, influenza A, fluid shear stress and atherosclerosis, the P3K-Akt signaling pathway, the HIF-1 signaling pathway, coronavirus disease (COVID-19), measles, human papillomavirus infection, EGFR tyrosine kinase inhibitor resistance, the IL-17 signaling pathway, apoptosis, and alcoholic liver disease.

[0134] 2.5 Composition-Target Molecular Docking

[0135] In molecular docking, the lower the energy of receptor-ligand binding, the greater the likelihood of spontaneous binding. Molecular docking results showed that five key components of Qingxuan Jiedu granules—quercetin, luteolin, kaempferol, beta-sitosterol, and troxerutin—had binding energies <0 kJ·mol⁻¹ to key COVID-19-related proteins TNF, STATE3, IL6, EGFR, and AR. -1 This indicates that the key components of Qingxuan Jiedu Granules have good binding activity with proteins that regulate COVID-19. The molecular docking results are shown in Table 1. Figure 6 , Figure 7 , Figure 8 , Figure 9 .

[0136] Table 1 Molecular docking results

[0137]

[0138]

[0139] In summary, this invention demonstrates, through the construction of a "drug-component-intersection target" relationship and the analysis of the mechanism of action of each effective component with viral targets, that the synergistic effect of each effective component can effectively target the common cold virus and the novel coronavirus, thus making the target of action of the traditional Chinese medicine composition precise, the mechanism of action precise, and the therapeutic effect precise.

[0140] Example 3: Pharmacological Study of Qingxuan Jiedu Granules

[0141] 1. Clinical application of Qingxuan Jiedu Granules - Treatment of mild cases of novel coronavirus

[0142] 1.1 Inclusion criteria

[0143] (1) Clinical manifestations: one of the following symptoms: sore throat, cough, fever, nasal congestion, runny nose, fatigue, etc.

[0144] (2) Body temperature <39℃;

[0145] (3) Blood routine test: WBC ≤ 10 × 10 12 / L, N≤0.75;

[0146] (4) Age 18 to 80 years old, gender not limited.

[0147] 1.2 Exclusion Criteria

[0148] (1) Upper respiratory tract bacterial infection;

[0149] (2) Pneumonia;

[0150] (3) Breastfeeding, pregnancy, or desire to become pregnant;

[0151] (4) Patients with abdominal pain or diarrhea.

[0152] 1.3 Usage and Dosage

[0153] (1) One packet of Chinese herbal decoction, taken orally twice daily;

[0154] (2) Treatment course: 3 to 5 days.

[0155] 1.4 Observation Indicators

[0156] (1) Observation of therapeutic effect: symptoms such as sore throat, cough, fever, nasal congestion, runny nose, and fatigue disappeared, improved, did not improve, or worsened;

[0157] (2) Adverse events: diarrhea, palpitations, etc.

[0158] 1.5 Follow-up

[0159] Follow-up methods: Hospital visits or WeChat follow-up.

[0160] Follow-up period: Days 1, 2, 3, 4, and 5 of medication.

[0161] 1.6 Follow-up Results

[0162] The follow-up results of one case are shown in Table 2.

[0163] Table 2 Clinical observation and follow-up table of Qingxuan Jiedu formula

[0164]

[0165] 1.7 Research Results

[0166] A total of 77 patients participated in the clinical efficacy observation of Qingxuan Jiedu Granules. Among them, 2 patients refused to take the Chinese medicine after enrollment, and 2 patients stopped taking the Chinese medicine due to diarrhea. In fact, 73 patients completed the clinical observation. All of them were mild cases of novel coronavirus infection. Among them, there were 44 male patients and 29 female patients. The average age was 44 years old, the youngest age was 19 years old, and the oldest age was 79 years old. The follow-up form of one patient is shown in Table 2.

[0167] Regarding efficacy: The effects of Qingxuan Jiedu Granules on improving patients' sore throat, runny nose, cough, nasal congestion, fatigue, and body temperature were observed. Of the 25 patients with sore throat, 19 experienced relief after treatment, 5 showed improvement, and 1 showed no improvement, resulting in a symptom relief rate of 96.00%. Of the 22 patients with runny nose, 16 experienced relief after treatment, 6 showed improvement, resulting in a symptom relief rate of 100%. Of the 55 patients with cough, 19 experienced relief after treatment, 34 showed improvement, and 2 showed no improvement, resulting in a symptom relief rate of 96.36%. Of the 17 patients with nasal congestion, 10 experienced relief after treatment, 6 showed improvement, and 1 showed no improvement, resulting in a symptom relief rate of 94.11%. Of the 12 patients with fatigue, 4 experienced relief after treatment, 7 showed improvement, and 1 showed no improvement, resulting in a symptom relief rate of 91.67%. Of the 7 patients with fever, 6 had their temperatures return to normal after medication, while 1 still had a low-grade fever.

[0168] Regarding safety: During the study, two patients experienced diarrhea after taking the medication and subsequently refused to continue taking it. Nineteen patients have not yet completed clinical observation, which will be completed in the future.

[0169] 2. Clinical observation of Qingxuan Jiedu formula for the treatment of viral colds

[0170] 2.1 Inclusion criteria are the same as 1.1

[0171] 2.2 Exclusion criteria are the same as 1.2

[0172] 2.3 Usage and dosage are the same as 1.3

[0173] 2.4 The observation indicators are the same as in 1.4.

[0174] 2.5 Follow-up is the same as 1.5

[0175] 2.6 Follow-up Results

[0176] The follow-up results of 45 cases are shown in Table 3.

[0177] Table 3 Clinical observation and follow-up table of Qingxuan Jiedu formula

[0178]

[0179]

[0180]

[0181] 2.7 Research Results

[0182] A total of 45 patients participated in the clinical efficacy observation of Qingxuan Jiedu Granules. All of them were patients with viral colds, including 31 male patients and 14 female patients. The average age was 29 years old, the youngest age was 17 years old, and the oldest age was 58 years old. The follow-up data of 45 cases are shown in Table 3.

[0183] Regarding efficacy: The effects of Qingxuan Jiedu granules on improving patients' sore throat, runny nose, cough, and body temperature were observed. A total of 45 people used Qingxuan Jiedu granules, of whom 30 / 45 had fever, 17 / 45 had sore throat, 6 / 45 had runny nose, and 14 / 45 had cough. 16 / 45 used it in combination with antipyretics, and 19 / 45 used it in combination with antibiotics. All patients with symptoms such as sore throat, runny nose, and cough (except for one patient who improved after 72 hours) showed improvement within 48 hours after taking the medication. Of the 30 patients with fever, 27 / 30 recovered to normal within 24 hours, and 2 patients had their body temperature return to normal after 48 hours.

[0184] Example 4: Quality Study of Qingxuan Jiedu Granules - Thin-Layer Chromatography Detection

[0185] 1. Thin-layer chromatography detection of raw ephedra

[0186] (1) Preparation of test solution

[0187] Dissolve 10g of Qingxuan Jiedu granules in water, adjust the pH to 11-12 with sodium hydroxide test solution, extract twice with dichloromethane, 20mL each time, combine the dichloromethane extracts, evaporate to dryness, dissolve the residue in 2mL of methanol to obtain the test solution.

[0188] (2) Preparation of negative test samples

[0189] Prepare a negative test sample containing a drug composition that does not contain raw ephedra according to step (1).

[0190] (3) Preparation of reference solution

[0191] Take raw ephedra reference powder, add 50 mL of water and reflux for 1 hour, cool, filter, and use as raw ephedra reference solution.

[0192] (4) Thin-layer chromatography test

[0193] Take 10 μL of each of the above three solutions and spot them separately on the same silica gel G thin-layer plate. Use dichloromethane-methanol-concentrated ammonia solution (volume ratio of 20:5:0.5) as the developing solvent, develop, remove, air dry, spray with 2% ninhydrin solution, and heat at 105℃ until the spots are clearly visible. In the chromatogram of the test sample, the same purplish-red spots appear at the corresponding positions as in the chromatogram of the reference medicinal material. No interference was observed in the negative test.

[0194] (5) Durability testing

[0195] Different temperatures: The thin-layer chromatography (TLC) development of raw ephedra was tested at 10℃ and 22℃ using the above method. Figure 10 The results showed no significant difference in the chromatograms at the two temperatures. Therefore, this method is suitable for thin-layer chromatographic identification of raw ephedra in Qingxuan Jiedu granules at different temperatures. Figure 10 (b) Figure 10 (c), where Figure 10 In (b), 1 represents the ephedra reference material. 2, 3, and 4 represent different batches of test samples. Figure 10 In (c), 1 is the ephedra reference material. 2, 3, and 4 are different batches of test samples.

[0196] Different humidity levels: The feasibility of thin-layer chromatography (TLC) identification of raw ephedra in Qingxuan Jiedu granules was tested at 30% and 60% humidity. Results showed no significant difference in chromatograms between the two humidity levels, with clear TLC identification points in both cases. Therefore, this method is suitable for TLC identification of raw ephedra under different humidity conditions. (See attached image) Figure 10 (d) Figure 10 (e), its Figure 10 In (d), 1 is the raw ephedra reference material, and 2, 3, and 4 are different batches of test samples; Figure 10 In (e), 1 is raw ephedra reference material, and 2, 3, and 4 are different batches of test samples.

[0197] Thin-layer chromatography plates from different manufacturers: Development was performed using Qingdao Marine silica gel G plates (batch number: 20200809) and Yantai Huanghai thin-layer chromatography silica gel G plates (batch number: 20210109). No significant differences were observed in the results, indicating that this method is well-suited to different manufacturers and brands of thin-layer chromatography plates. (See [link to relevant documentation]). Figure 10 (f), 10(g), Figure 10 (f) and 10(g) 1 is raw ephedra reference material, and 2, 3 and 4 are different batches of test samples.

[0198] 2. Thin-layer chromatography detection of raw licorice

[0199] (1) Preparation of test solution

[0200] Dissolve 10g of Qingxuan Jiedu Granules in 2ml of hydrochloric acid, heat in a water bath at 100℃ for 1 hour, remove, cool, filter, extract the filtrate three times with 30mL of water-saturated n-butanol each time, combine the n-butanol extracts, evaporate to dryness in a water bath, dissolve the residue in 2mL of methanol to obtain the test solution.

[0201] (2) Preparation of negative test samples

[0202] Prepare a negative test sample that does not contain raw licorice drug composition according to step (1).

[0203] (3) Preparation of reference solution

[0204] Take raw licorice reference material powder, add 100mL of water and reflux for 30 minutes, filter, add 3mL of hydrochloric acid to the filtrate, heat in a water bath at 100℃ for 1 hour, remove, cool, filter, extract the filtrate three times with 30mL of water-saturated n-butanol each time, combine the n-butanol extracts, evaporate to dryness, add 2mL of methanol to dissolve the residue, and use as the raw licorice reference material solution.

[0205] (4) Thin-layer chromatography test

[0206] Take 10 μL of each of the above three solutions and spot them separately on the same silica gel G thin-layer plate. Use petroleum ether (60-90℃)-toluene-ethyl acetate-glacial acetic acid (volume ratio of 10:20:7:0.5) as the developing solvent, develop, remove, air dry, spray with 10% sulfuric acid ethanol solution, and heat at 105℃ until the spots are clearly visible. In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatogram of the reference medicinal material. No interference was observed in the negative test.

[0207] (5) Durability testing

[0208] Different temperatures: The development of licorice root under thin-layer chromatography (TLC) at 4℃ and 25℃ was tested using the above method. Figure 11 The results showed no significant difference in the chromatograms at the two temperatures; therefore, this method is suitable for thin-layer chromatography identification of raw licorice in Qingxuan Jiedu granules at different temperatures. Figure 11 (b) Figure 11 (c), its Figure 11 (b) Figure 11 In (c), 1 is raw licorice reference material, and 2, 3, and 4 are different batches of test samples.

[0209] Different humidity levels: The feasibility of a thin-layer chromatography (TLC) method for identifying raw licorice in Qingxuan Jiedu granules under 30% and 60% humidity was tested. Results showed no significant difference in the chromatograms under both humidity levels, with clear and distinct TLC identification points. Therefore, this method is suitable for the TLC identification of raw licorice under different humidity levels. (See [link to relevant documentation]). Figure 11 (d) Figure 11 (e), its Figure 11 (d) Figure 11 In (e), 1 is the licorice reference material, and 2, 3, and 4 are different batches of test samples.

[0210] Thin-layer chromatography plates from different manufacturers: Development was performed using Qingdao Marine silica gel G plates (batch number: 20200809) and Sinopharm Chemical Research thin-layer chromatography silica gel G plates (batch number: 20210109). No significant differences were observed in the results, indicating that this method is well-suited to different manufacturers and brands of thin-layer chromatography plates. (See [link to relevant documentation]). Figure 11 (f) Figure 11 (g), where Figure 11 (f) Figure 11In (g), 1 is the licorice reference material, and 2, 3, and 4 are different batches of tested materials.

[0211] 3. Thin-layer chromatography detection of Forsythia

[0212] (1) Preparation of test solution

[0213] Dissolve the drug composition in 30 mL of methanol, sonicate for 30 min, filter, evaporate the filtrate to dryness, and dissolve the residue in 1 mL of methanol to obtain the test solution.

[0214] (2) Preparation of negative test solution

[0215] Take a drug composition that does not contain Forsythia and prepare a negative test solution according to the test solution preparation method.

[0216] (3) Preparation of control herbal solution

[0217] Take 3g of Forsythia suspensa reference herb powder, add 10mL of methanol, and prepare the reference herb solution according to the method for preparing the test solution.

[0218] (4) Thin-layer chromatography test

[0219] Take 20 μL of each of the above three solutions and spot them on the same silica gel G thin-layer plate. Use chloroform-methanol (volume ratio 49:1) as the developing solvent, develop, remove, air dry, spray with 10% sulfuric acid ethanol solution, bake at 105℃ for 5 minutes, place under UV 366nm, and develop color. In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatogram of the reference medicinal material, and no interference is observed in the negative test.

[0220] (5) Durability testing

[0221] Different temperatures: The development of Forsythia suspensa thin-layer identification using the above method was tested at 10℃ and 23℃ respectively. Figure 12 The chromatograms at the two temperatures showed no significant difference, therefore this method is suitable for thin-layer chromatographic identification of Forsythia suspensa in Qingxuan Jiedu Granules at different temperatures. (See...) Figure 12 (b) Figure 12 (c), where Figure 12 In (b), 1 represents the Forsythia suspensa control material. 2, 3, and 4 represent different batches of test samples. Figure 12 In (c), 1 is the Forsythia suspensa control material. 2, 3, and 4 are different batches of test samples.

[0222] Different humidity levels: The feasibility of the thin-layer chromatography (TLC) method for identifying Forsythia suspensa in Qingxuan Jiedu granules was tested at 30% and 60% humidity. The results are shown in the figure below. There was no significant difference in the chromatograms under the two humidity levels, and both showed clear TLC identification points. Therefore, this method is suitable for the TLC identification of Forsythia suspensa under different humidity levels. Figure 12 (d) Figure 12 (e), its Figure 12In (d), 1 represents a negative result, 2 represents the Forsythia suspensa control material, and 3, 4, and 5 represent different batches of test samples. Figure 12 In (e), 1 represents a negative result, 2 represents the Forsythia suspensa control material, and 3, 4, and 5 represent different batches of test samples.

[0223] Thin-layer chromatography plates from different manufacturers: Development was performed using Qingdao Marine silica gel G plates (batch number: 20200809) and Yantai Huanghai thin-layer chromatography silica gel G plates (batch number: 20210109). No significant differences were observed in the results, indicating that this method is well-suited to different manufacturers and brands of thin-layer chromatography plates. (See [link to relevant documentation]). Figure 12 (f) Figure 12 (g), where Figure 12 (f) Figure 12 In (g), 1 represents a negative result, 2 represents the Forsythia suspensa control material, and 3, 4, and 5 represent different batches of test samples.

[0224] 4. Thin-layer chromatography detection of Codonopsis pilosula

[0225] (1) Preparation of test solution

[0226] Dissolve the drug composition in 30 mL of methanol, wet soak, sonicate for 30 min, filter, evaporate to dryness, and dissolve the residue in 1 mL of methanol to obtain the test solution.

[0227] (2) Preparation of negative test solution

[0228] Take a drug composition that does not contain Codonopsis pilosula and prepare a negative test solution according to the test solution preparation method.

[0229] (3) Preparation of control herbal solution

[0230] Take 3g of reference medicinal material powder, add 10mL of methanol, and prepare the reference medicinal material solution according to the method for preparing the test solution.

[0231] (4) Thin-layer chromatography test

[0232] Take 5 μL of each of the above three solutions and spot them on the same silica gel G thin-layer plate. Use n-butanol-glacial acetic acid-water as the developing solvent (volume ratio 4:1:1) for 15 minutes to pre-saturate the plate, develop the solution, remove the plate, air dry it, spray it with 0.2% ninhydrin solution, and heat it at 105℃ until the spots are clearly visible. In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatogram of the reference medicinal material. No interference was observed in the negative test.

[0233] (5) Durability testing

[0234] Different temperatures: The development of the Pseudostellaria heterophylla thin-layer chromatography (TLC) for identification using the above method was tested at 10℃ and 23℃ respectively. For example... Figure 13 The chromatograms at the two temperatures showed no significant difference, therefore this method is suitable for thin-layer chromatographic identification of Codonopsis pilosula in Qingxuan Jiedu Granules at different temperatures. (See...) Figure 13 (b) Figure 13 (c), where Figure 13 In (b), 1 represents the reference material of Codonopsis pilosula, and 2, 3, and 4 represent different batches of test samples. Figure 13 In (c), 1 is the reference material of Codonopsis pilosula, and 2, 3, and 4 are different batches of test samples.

[0235] Different humidity levels: The feasibility of thin-layer chromatography (TLC) identification of Codonopsis pilosula in Qingxuan Jiedu granules was tested at 30% and 60% humidity. The results are shown in the figure below. There was no significant difference in the chromatograms under the two humidity levels, and both showed clear TLC identification points. Therefore, this method is suitable for TLC identification of Codonopsis pilosula under different humidity levels. Figure 13 (d) Figure 13 (e), its Figure 13 In (d), 1 represents a negative result, 2 represents the reference material of Codonopsis pilosula, and 3, 4, and 5 represent different batches of test samples; Figure 13 In (e), 1 represents a negative result, 2 represents the reference material of Codonopsis pilosula, and 3, 4, and 5 represent different batches of test samples.

[0236] Thin-layer chromatography plates from different manufacturers: Development was performed using Qingdao Marine silica gel G plates (batch number: 20200809) and Yantai Huanghai thin-layer chromatography silica gel G plates (batch number: 20210109). No significant differences were observed in the results, indicating that this method is well-suited to different manufacturers and brands of thin-layer chromatography plates. (See [link to relevant documentation]). Figure 13 (f) Figure 13 (g), Figure 13 (f) Figure 13 In (g), 1 represents a negative result, 2 represents the reference material of Codonopsis pilosula, and 3, 4, and 5 represent different batches of test samples.

[0237] Example 5: Quality Study of Qingxuan Jiedu Granules - High Performance Liquid Chromatography

[0238] 1. Chromatographic conditions for high performance liquid chromatography (HPLC)

[0239] An Agilent Zorbax SB-Phenyl column (250 nm × 4.6 mm, 5 μm) was used for elution with acetonitrile as mobile phase A and 0.2% phosphoric acid aqueous solution as mobile phase B. The column temperature was 40 °C, the detection wavelength was 210 nm, the injection volume was 10 μL, and the theoretical plate number was set to be higher than 3000 for ephedrine hydrochloride. The elution program is shown in Table 4.

[0240] Table 4 Elution Procedure

[0241] time min Mobile phase A% Mobile phase B% Flow rate mL / min 0 3 97 1 30 3 97 1 35 95 5 1 45 95 5 1 50 3 97 1

[0242] 2. Preparation of reference solution

[0243] Accurately weigh 0.0308 g of 100% pure ephedrine hydrochloride standard and 0.0238 g of 99.8% pure pseudoephedrine hydrochloride standard, and dissolve them in anhydrous methanol to prepare reference solutions containing 3.08 mg / mL ephedrine hydrochloride and 2.38 mg / mL pseudoephedrine hydrochloride.

[0244] 3. Preparation of the test solution

[0245] Dissolve 10g of the drug composition in 100mL of water, add 120mL of 20% sodium hydroxide solution and 7.5g of sodium chloride, distill, collect the distillate, add 5mL of 0.5mol / L hydrochloric acid solution beforehand, collect 120mL, add water to make up to volume, shake well, filter through a 0.45μm microporous membrane, and take the filtrate to obtain the test solution; take the drug composition that does not contain ephedrine, and prepare the negative test solution according to the test solution preparation method;

[0246] 4. System adaptability test

[0247] Accurately pipette 10 μL each of the reference solution and the test solution into the liquid chromatograph and determine their absorption. The test solution and the reference solution show significant absorption at the same position. The theoretical plate number, calculated based on the paeoniflorin peak, is greater than or equal to 3000.

[0248] Take appropriate amounts of reference standards, mix well, and dilute 2, 4, 8, 16, 88, 176, and 704 times to prepare reference standard solutions of different concentrations. Determine the concentrations under the chromatographic conditions described above. Plot standard curves with concentration on the x-axis and peak area on the y-axis. The regression equations for ephedrine hydrochloride and pseudoephedrine hydrochloride are y = 9353.6x - 4.2432 (r = 0.9997, n = 8), and for pseudoephedrine hydrochloride, y = 10876x - 29.694 (r = 0.9997, n = 8). The results show that ephedrine hydrochloride and pseudoephedrine hydrochloride exhibit good linearity in the ranges of 0.0544–15.7286 mg / mL and 0.0798–11.945 mg / mL, respectively.

[0249] 5. Sample content determination

[0250] Three batches of Qingxuan Jiedu granules, prepared according to the method in Example 1, were precisely measured in two portions. Qingxuan Jiedu solutions were prepared according to the above requirements, and 10 μL of each solution was injected under the above chromatographic conditions for determination and calculation. The results of the determination of ephedrine and pseudoephedrine content in the three batches of Qingxuan Jiedu granules are shown in Table 5. The experimental results show good reproducibility between batches.

[0251] Table 5 Content Results

[0252] batch Ephedrine hydrochloride (mg / mL) Pseudoephedrine hydrochloride (mg / mL) Total amount (mg / mL) 1 0.1154 0.1464 0.2617 2 0.1162 0.1469 0.2631 3 0.1132 0.1517 0.2649

Claims

1. A traditional Chinese medicine composition for treating mild cases of COVID-19 or influenza, characterized in that, By weight, it is made from 10 parts mulberry leaves, 10 parts chrysanthemum, 10 parts forsythia, 10 parts honeysuckle, 6 parts platycodon, 10 parts bitter almond, 10 parts honey loquat leaves, 3 parts ephedra, 8 parts bupleurum, 12 parts verbena, 15 parts reed rhizome, 10 parts codonopsis, and 6 parts licorice.

2. A pharmaceutical composition, characterized in that, The pharmaceutical composition is made from an extract of the traditional Chinese medicine composition of claim 1 and a pharmaceutically acceptable carrier, wherein the pharmaceutical composition is in the form of granules, capsules or tablets.

3. A method for preparing the pharmaceutical composition according to claim 2, characterized in that, Includes the following steps: (1) Combined extraction: Take mulberry leaves, chrysanthemum, forsythia, honeysuckle, platycodon, bitter almond, honey loquat leaf, ephedra, bupleurum, verbena, reed rhizome, codonopsis and licorice by weight, combine them and extract with water, filter the obtained extract, concentrate and centrifuge to obtain the centrifuged solution; (2) Prepare a pharmaceutical composition by adding a pharmaceutically acceptable carrier.

4. The preparation method according to claim 3, characterized in that, When the drug composition prepared in step (2) is granules, dextrin is placed in a fluidized bed. When the material temperature rises, the centrifuged solution obtained in step (1) is spray-dried to obtain granules.

5. A method for quality testing of the pharmaceutical composition according to claim 2, characterized in that, The assay includes thin-layer chromatography (TLC) detection of ephedra, licorice, forsythia, and codonopsis. Specifically, the TLC developing solvent for ephedra is dichloromethane-methanol-concentrated ammonia solution (v / v ratio 20:5:0.5); for licorice, it is petroleum ether-toluene-ethyl acetate-glacial acetic acid solution (v / v ratio 10:20:7:0.5); for forsythia, it is chloroform-methanol solution (v / v ratio 49:1); and for codonopsis, it is n-butanol-glacial acetic acid-water solution (v / v ratio 4:1:1).

6. The method for quality testing of the pharmaceutical composition according to claim 5, characterized in that, It also includes high-performance liquid chromatography (HPLC) detection, with the specific steps as follows: (1) Preparation of reference solution: Take ephedrine hydrochloride standard and pseudoephedrine hydrochloride standard and prepare reference solution; (2) Preparation of test solution: Dissolve the drug composition in water, add sodium hydroxide solution and sodium chloride, distill, collect the distillate, add hydrochloric acid solution beforehand, collect, add water to make up to volume, shake well, filter, and take the filtrate to obtain the test solution; take the drug composition that does not contain ephedrine and prepare the negative test solution according to the test solution preparation method; (3) Plotting the standard curve: Take the reference standard, mix and dilute it to prepare reference standard solutions of different concentrations, and plot the standard curve with concentration as the abscissa x and peak area as the ordinate y. The regression equation of ephedrine hydrochloride is y=9353.6x-4.2432, and the regression equation of pseudoephedrine hydrochloride is y=10876x-29.

694. (4) Detection: The reference solution prepared in step (1) and the test solution prepared in step (2) were respectively injected into the high performance liquid chromatograph. The chromatographic conditions were phenyl column, column temperature: 40℃, detection wavelength: 210nm, acetonitrile as mobile phase A and 0.2% phosphoric acid aqueous solution as mobile phase B for elution. The elution program was as follows: 0~30min, A:B maintained at 3:97, 30~35min, A:B converted to 95:5, 35~45min, A:B maintained at 95:5, 45~50min, A:B converted to 3:

97. (5) Calculate the content: Based on the standard curve equation in step (3), calculate the content of ephedrine and pseudoephedrine in the test solution.

7. The method for quality testing of the pharmaceutical composition according to claim 6, characterized in that, In step (3), the detection linear range of ephedrine hydrochloride is 0.0544~15.7286 mg / mL, and the detection linear range of pseudoephedrine hydrochloride is 0.0798~11.945 mg / mL.

8. The use of the traditional Chinese medicine composition of claim 1 or the pharmaceutical composition of claim 2 in the preparation of a drug for treating mild COVID-19 or influenza.

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