Composition with sore-throat relieving, cough relieving and antiviral effects as well as preparation method and application thereof

Abstract

The invention provides a composition with sore-throat relieving, cough relieving and antiviral effects as well as a preparation method and application thereof, and belongs to the technical field of pharmaceutical compositions. The composition provided by the invention comprises 11-20 parts of fermented soybean, 22-26 parts of medicated leaven, 1-3 parts of blackberry lily, 2-4 parts of platycodon grandiflorum, 5-8 parts of perilla leaves, 32-35 parts of fructus aurantii, 1-3 parts of coptis chinensis, 4-8 parts of trichosanthes kirilowii maxim, 5-9 parts of loquat leaves, 2-4 parts of scutellaria baicalensis, 1-2 parts of cape jasmine, 5-9 parts of mangnolia officinalis, 4-8 parts of radix bupleuri, 12-16 parts of rhizoma pinelliae preparata, 5-8 parts of radix curcumae, 12-18 parts of liquorice, 4-6 parts of fresh ginger, 32-36 parts of Chinese dates and 4-8 parts of poria cocos. On the basis of adjusting the monarch, minister, assistant and guide relationship, the composition disclosed by the invention is combined with the improvement of the preparation method, so that efficient extraction of components, synergistic enhancement of the drug effect and process optimization are realized, and finally the remarkable technical effects of relieving sore throat and cough, and resisting viruses and mycoplasma pneumoniae are achieved.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical compositions, and in particular relates to a composition having both throat-relieving, cough-relieving and antiviral effects, as well as a preparation method and application thereof. Background Art

[0002] Respiratory diseases are a type of disease with a high incidence rate worldwide, including pharyngitis, bronchitis, pneumonia, etc. Among them, respiratory diseases caused by viruses (such as influenza virus, respiratory syncytial virus) and Mycoplasma pneumoniae infections are highly contagious, have a prolonged course, and are prone to recurrence, posing a serious threat to public health.

[0003] Currently, treatments for these diseases primarily include chemical drugs and traditional Chinese medicines. While chemical drugs such as neuraminidase inhibitors (such as oseltamivir) and antibiotics like azithromycin have demonstrated efficacy against specific pathogens, they also have numerous limitations. For example, oseltamivir is only effective against influenza viruses, ineffective against other viruses and Mycoplasma pneumoniae, and long-term use can easily induce viral resistance. While antibiotics like azithromycin have inhibitory effects against Mycoplasma pneumoniae, they can easily cause adverse reactions such as gastrointestinal disturbances and liver damage. Furthermore, overuse can exacerbate bacterial resistance, making treatment more difficult.

[0004] Traditional Chinese medicine preparations offer the advantages of multiple targets and minimal side effects in the treatment of respiratory diseases. Some existing TCM compositions can achieve single efficacy, such as soothing and antitussive treatment or antiviral treatment. However, TCM compositions that simultaneously possess the triple efficacy of soothing and antitussive treatment, antiviral treatment, and anti-Mycoplasma pneumoniae treatment remain relatively scarce. The main technical difficulties lie in: first, the precise combination of active ingredients with different mechanisms of action is required to avoid mutual antagonism. For example, antiviral ingredients may compete with anti-Mycoplasma pneumoniae ingredients for dissolution. Second, the pathogenic mechanisms of respiratory viruses and Mycoplasma pneumoniae are complex. Viruses can invade cells through surface protein adsorption, while Mycoplasma pneumoniae relies on specific adhesion factors to adhere to the respiratory epithelium. Therefore, drugs must act simultaneously on multiple pathways to achieve synergistic treatment. Furthermore, the preparation process of existing TCM preparations often suffers from low extraction rates of active ingredients and poor stability, making it difficult to achieve the desired triple efficacy simultaneously. Therefore, developing a safe, efficient, and multi-functional TCM composition has become a pressing technical challenge. Summary of the Invention

[0005] In view of this, one of the objects of the present invention is to provide a composition having the effects of relieving throat and cough, antiviral and anti-Mycoplasma pneumoniae.

[0006] A second object of the present invention is to provide a method for preparing the above composition.

[0007] The third object of the present invention is to provide the use of the above composition in the preparation of anti-Mycoplasma pneumoniae products.

[0008] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions: The invention provides a composition with both soothing and antitussive effects and antiviral effects. The composition comprises the following raw materials in parts by weight: 11-20 parts of light fermented black beans, 22-26 parts of Shenqu, 1-3 parts of Belamcanda Root, 2-4 parts of Platycodon Grandiflorum, 5-8 parts of Perilla Leaf, 32-35 parts of Citrus Aurantium, 1-3 parts of Coptis Rhizome, 4-8 parts of Trichosanthes Fructus, 5-9 parts of Loquat Leaf, 2-4 parts of Scutellaria Baicalensis, 1-2 parts of Gardenia Fructus, 5-9 parts of Magnolia Bark, 4-8 parts of Bupleurum Root, 12-16 parts of Pinellia Tuber, 5-8 parts of Curcuma Radix, 12-18 parts of Licorice Root, 4-6 parts of Ginger, 32-36 parts of Jujube and 4-8 parts of Poria.

[0009] Preferably, the antiviral agent includes anti-respiratory syncytial virus or anti-influenza virus.

[0010] The present invention also provides a method for preparing the above composition, comprising the following steps: Mixing coptis root, scutellaria root, magnolia bark and ginger to obtain mixture 1, crushing the mixture 1 and mixing it with rice wine to moisten it, and steaming the moistened medicine for 30 minutes to 60 minutes to obtain a steamed product; Mix light fermented black beans, Shenqu, Belamcanda chinensis, Platycodon grandiflorum, Perilla frutescens leaves and Citrus aurantium to obtain mixture 2, mix mixture 2 with water, soak, and boil to obtain a decoction; Mix and grind Trichosanthes kirilowii, loquat leaves, bupleurum and Poria cocos to obtain a mixture 3, mix the mixture 3 with a 60% to 80% by volume ethanol solution, perform reflux extraction, and collect the extract; Gardenia jasminoides, pinellia tuber, turmeric, liquorice and jujube are mixed and crushed to obtain mixture 4, the mixture 4 is mixed and moistened with a percolation solvent, and then the percolation solvent is added for impregnation. After the impregnation is completed, percolation is started, and the percolation liquid is collected to a volume of 2 to 4 times the amount of mixture 4; the percolation solvent is an ethanol solution with a volume fraction of 45% to 60%; The decoction, the extract and the percolated liquid are mixed and concentrated to a relative density of 1.1 to 1.2 to obtain a concentrate; the concentrate is mixed with the steamed product to obtain the composition.

[0011] Preferably, the powder is sieved through a 20-40 mesh sieve after being crushed.

[0012] Preferably, the weight ratio of the mixture 1 to the rice wine is 10:2-3; and the steeping time is 2h-2.5h.

[0013] Preferably, the weight ratio of the mixture 2 to water is 1:7-10; the soaking time is 20 min-30 min; and the decocting time is 1 h-1.5 h.

[0014] Preferably, the weight ratio of the mixture 3 to the ethanol solution with a volume fraction of 60% to 80% is 1:6 to 8; the number of reflux extractions is 1 to 2 times, and the time for each reflux extraction is 0.5 h to 1 h.

[0015] Preferably, during wetting, the weight ratio of the mixture 4 to the diafiltration solvent is 1:2~3, and the wetting time is 1h~2h; during impregnation, the weight of the added diafiltration solvent is 4~6 times the weight of the mixture 4, and the impregnation time is 8h~10h; the diafiltration speed is 3~5mL per minute.

[0016] The present invention also provides the use of the above composition or the above preparation method in preparing an anti-Mycoplasma pneumoniae product.

[0017] Beneficial effects of the present invention: The composition of the present invention adjusts the dosage ratio of each medicinal flavor, takes light fermented black beans and Shenqu as the main medicine, takes Belamcanda chinensis, Platycodon grandiflorum, Perilla leaf and Citrus aurantium as the ministerial medicine, takes Coptis chinensis, Scutellaria baicalensis, Gardenia jasminoides, Trichosanthes kirilowii, Loquat leaf, Magnolia officinalis, Bupleurum chinense, Pinellia ternata, Curcuma aromatica and Poria cocos as the adjuvant medicine, and takes Licorice, ginger and jujube as the guiding medicine. Through the rigorous compatibility of the main, minister, adjuvant and guiding medicines, the various medicinal materials work together to form an integrated treatment method of "dispelling exterior pathogens - regulating Qi - clearing away heat and detoxifying - strengthening the body and consolidating the foundation", and jointly achieves the effects of relieving throat and cough and antiviral, while taking into account multiple functions such as regulating the spleen and stomach, regulating Qi and resolving phlegm, clearing away heat and detoxifying, etc., can not only directly inhibit viruses and alleviate respiratory inflammation, but also indirectly fight diseases by regulating the body's immunity. Compared with drugs or ingredients with single functions, the composition of the present invention has the advantages of being more comprehensive and efficient in treating respiratory infectious diseases. Furthermore, the present composition focuses on holistically regulating the body's condition. Herbs such as Shenqu (Chinese Medicinal Herb), Poria cocos, and licorice strengthen the spleen and stomach, enhancing acquired health. Ginger and jujube harmonize the Ying and Wei systems, improving immune function. This holistic approach not only treats current ailments but also enhances the body's resistance to disease, reducing the recurrence of respiratory illnesses and achieving the dual effects of both treatment and prevention.

[0018] The composition of the present invention adjusts the relationship between monarch, minister, adjuvant and envoy, combines the improvement of the preparation method, and realizes efficient extraction of ingredients, synergistic enhancement of drug efficacy and process optimization through precise step-by-step processing according to the characteristics and efficacy differences of the medicinal materials, and ultimately achieves the remarkable technical effects of not only relieving throat and cough, but also resisting viruses and Mycoplasma pneumoniae. DETAILED DESCRIPTION

[0019] The invention provides a composition with both soothing and antitussive effects and antiviral effects. The composition comprises the following raw materials in parts by weight: 11-20 parts of light fermented black beans, 22-26 parts of Shenqu, 1-3 parts of Belamcanda Root, 2-4 parts of Platycodon Grandiflorum, 5-8 parts of Perilla Leaf, 32-35 parts of Citrus Aurantium, 1-3 parts of Coptis Rhizome, 4-8 parts of Trichosanthes Fructus, 5-9 parts of Loquat Leaf, 2-4 parts of Scutellaria Baicalensis, 1-2 parts of Gardenia Fructus, 5-9 parts of Magnolia Bark, 4-8 parts of Bupleurum Root, 12-16 parts of Pinellia Tuber, 5-8 parts of Curcuma Radix, 12-18 parts of Licorice Root, 4-6 parts of Ginger, 32-36 parts of Jujube and 4-8 parts of Poria.

[0020] In the present invention, the amount of the light fermented black beans is preferably 13-17 parts, more preferably 14-16 parts; the amount of the Shenqu is preferably 23-25 ​​parts, more preferably 24 parts; the amount of the Belamcanda Root is preferably 2 parts; the amount of the Platycodon Grandiflorum is preferably 3 parts; the amount of the Perilla Leaf is preferably 6-7 parts; the amount of the Citrus Aurantium is preferably 33-34 parts; the amount of the Coptis chinensis is preferably 2 parts; the amount of the Trichosanthes Knotweed is preferably 5-7 parts, more preferably 6 parts; the amount of the loquat leaf is preferably 6-8 parts, more preferably 7 parts; the amount of the Scutellaria baicalensis is preferably 10-15 parts. The dosage is preferably 3 parts; the dosage of the Magnolia officinalis is preferably 6 to 8 parts, more preferably 7 parts; the dosage of the Bupleurum is preferably 5 to 7 parts, more preferably 6 parts; the dosage of the Pinellia ternata is preferably 13 to 15 parts, more preferably 14 parts; the dosage of the Curcuma aromatica is preferably 6 to 7 parts; the dosage of the Licorice is preferably 14 to 17 parts, more preferably 15 to 16 parts; the dosage of the ginger is preferably 5 parts; the dosage of the jujube is preferably 33 to 35 parts, more preferably 34 parts; the dosage of the Poria cocos is preferably 5 to 7 parts, more preferably 6 parts. The present invention has no special restrictions on the specific sources of the above-mentioned raw materials, and conventional commercial products in this field can be used. In the present invention, the antiviral preferably includes anti-respiratory syncytial virus or anti-influenza virus, and the anti-influenza virus preferably includes anti-influenza A virus.

[0021] In this composition, light fermented black beans have the effects of dispelling exterior pathogens, dissipating heat, and alleviating restlessness. They can dispel external pathogens, helping to alleviate symptoms such as fever and sore throat caused by exogenous pathogens, laying the foundation for treating respiratory infections. Shenqu (Shenqu) promotes digestion and soothes the stomach, regulating spleen and stomach function and strengthening the body's vital energy, helping to improve immunity and indirectly assist in antiviral treatment and alleviate respiratory symptoms. These two ingredients (light fermented black beans and Shenqu) serve as the main ingredients, working together to alleviate exterior pathogens and strengthen the body's vital energy, achieving the core effects of soothing throat, relieving coughs, and combating viruses.

[0022] In the composition of the present invention, Belamcanda chinensis clears away heat and detoxifies, eliminates phlegm and relieves sore throat, has an inhibitory effect on respiratory viruses, can reduce inflammation in the throat, relieve cough symptoms, and directly assist the main drug in achieving the effect of relieving sore throat and cough.

[0023] Platycodon grandiflorum clears the lungs, relieves sore throats, and dispels phlegm and pus, promoting the expulsion of phlegm from the respiratory tract and alleviating lung burden. Its lung-clearing properties help restore the normal function of lung qi, synergizing with Belamcanda Root to enhance cough and expectorant efficacy while also promoting respiratory function recovery during the antiviral process. Perilla leaf dispels exterior cold, promotes qi circulation and soothes the stomach, complementing light fermented black beans in clearing exterior cold and enhancing antiviral efficacy. Furthermore, by promoting qi circulation and soothing the stomach, it synergizes with Shenqu to regulate the spleen and stomach, strengthening the body's overall disease resistance. Citrus aurantium fructus regulates qi, relieves fullness, relieves stagnation, and alleviates bloating, promoting the smooth flow of qi, aiding the circulation and distribution of the drug, enhancing the effects of the monarch and ministerial herbs, and alleviating symptoms such as cough and chest tightness caused by stagnant qi in the respiratory tract. These herbs (Belamcanda Root, Platycodon grandiflorum, Perilla leaf, and Citrus aurantium fructus) collectively serve as ministerial herbs, complementing the monarch herbs to enhance the core efficacy of soothing sore throat, cough, and antiviral efficacy.

[0024] In the composition of the present invention, Rhizoma Coptidis, Radix Scutellariae, and Fructus Gardeniae are all heat-clearing and detoxifying drugs, which assist the monarch and minister drugs in enhancing the antiviral and anti-inflammatory effects, while clearing away heat and toxins in the body to prevent the condition from worsening. Fructus Trichosanthis clears heat and eliminates phlegm, relieves chest tightness and disperses stagnation, and Fructus Loquatis clears the lungs and relieves cough. Both can resolve phlegm and relieve cough, alleviate the symptoms of phlegm-heat in the lungs, and assist the monarch and minister drugs in relieving respiratory symptoms, and are particularly suitable for coughs accompanied by thick sputum. Magnolia officinalis can dry out dampness and eliminate phlegm, and relieve fullness. Together with Fructus Citrus Aurantii, it enhances the effect of regulating qi, regulates the respiratory qi mechanism, and simultaneously eliminates the symptoms such as chest tightness and fullness caused by phlegm-dampness obstruction, and assists the monarch and minister drugs in playing a role. Radix Bupleuri harmonizes the exterior and interior, soothes the liver and raises yang, can guide the various drugs into the meridians, harmonize the human qi mechanism, regulate the body's stress response during the antiviral process, enhance the overall therapeutic effect, and can also assist Fructus Sojae to resolve the exterior and dissipate heat. Pinellia ternata clears dampness and resolves phlegm, reduces adverse reactions and stops vomiting. It can eliminate phlegm and dampness in the respiratory tract, relieving symptoms such as cough and nausea. Its expectorant effect synergizes with other antitussive herbs, while also calming adverse reactions and soothing the stomach, alleviating gastrointestinal discomfort caused by illness or medication, complementing the main and auxiliary herbs. Curcuma zedoaria promotes blood circulation and relieves pain, promotes qi and relieves depression, and clears the heart and cools the blood. It can improve local circulation in the respiratory tract and help resolve inflammation. Its qi-promoting and depression-relieving effects synergize with those of Citrus aurantium and Magnolia officinalis, regulating qi and enhancing the overall therapeutic effect. It also assists the main and auxiliary herbs in clearing heat and toxins from the body. Poria cocos has a sweet and mild flavor and a neutral nature. It promotes diuresis and eliminates dampness, strengthens the spleen and calms the mind, and complements the main and auxiliary herbs in prescriptions in multiple ways. These herbs (Coptis chinensis, Scutellaria baicalensis, Gardenia jasminoides, Trichosanthes kirilowii, Loquat leaf, Magnolia officinalis, Bupleurum chinense, Pinellia ternata, Curcuma zedoaria, and Poria cocos) serve as adjuvants, coordinating the main and auxiliary herbs and balancing their multiple functions.

[0025] In the composition of the present invention, licorice nourishes the spleen and replenishes qi, clears away heat and detoxifies, and harmonizes the various medicinal ingredients. It not only replenishes qi and strengthens the body's immunity, but also harmonizes the properties and flavors of the various medicinal ingredients in the prescription, alleviating any potential bias between the medicinal ingredients, making the overall efficacy of the prescription more gentle and lasting. It also guides the medicinal ingredients directly to the affected area, better exerting their sore throat, cough, and antiviral effects. Ginger dispels cold, warms the middle and stops vomiting, while jujube nourishes the middle and replenishes qi, nourishes blood, and calms the mind. The combination of the two harmonizes the Ying and Wei, strengthens the spleen and stomach, promotes drug absorption and metabolism, and reduces gastrointestinal irritation from other drugs. The licorice harmonizes the various medicinal ingredients, allowing the prescription's efficacy to be better exerted. These medicinal ingredients (licorice, ginger, and jujube) collectively serve as guiding ingredients, harmonizing the various medicinal ingredients and guiding them to their meridians.

[0026] The present invention also provides a preparation method of the above-mentioned composition, comprising the following steps: mixing coptis root, scutellaria baicalensis, magnolia bark and ginger to obtain mixture 1, crushing the mixture 1 and mixing it with rice wine to moisten it, steaming the moistened medicine for 30 minutes to 60 minutes to obtain a steamed product; mixing light fermented black beans, Shenqu, Belamcanda Root, Platycodon grandiflorum, Perilla Leaf and Citrus aurantium to obtain mixture 2, mixing the mixture 2 with water, soaking it, decocting it and taking the decoction; mixing and crushing trichosanthes root, loquat leaf, bupleurum root and Poria cocos to obtain mixture 3, mixing the mixture 3 with 60% to 80% by volume of The steps of: mixing the mixture with an ethanol solution, performing reflux extraction, and collecting the extract; mixing and grinding gardenia, pinellia tuber, turmeric, liquorice, and jujube to obtain a mixture 4; mixing the mixture 4 with a percolation solvent to moisten the mixture; then adding the percolation solvent for impregnation; starting percolation after the impregnation, and collecting the percolation liquid to a volume 2 to 4 times that of the mixture 4; the percolation solvent is an ethanol solution with a volume fraction of 45% to 60%; mixing the decoction, the extract, and the percolation liquid, and concentrating the mixture to a relative density of 1.1 to 1.2 to obtain a concentrate; and mixing the concentrate with the steamed product to obtain the composition.

[0027] The present invention adopts differentiated extraction methods based on the compositional characteristics of different medicinal materials, significantly improving the yield of effective ingredients. Coptis chinensis, Scutellaria baicalensis, Magnolia officinalis, and ginger are mixed with rice wine and steamed. The rice wine promotes the dissolution of ingredients such as alkaloids and flavonoids. The steaming process also mitigates the bitter and cold nature of Coptis chinensis, reducing gastrointestinal irritation and preventing high temperatures from damaging heat-sensitive ingredients. Soaking and decocting herbs such as light fermented black beans and Shenqu effectively extracts water-soluble ingredients such as polysaccharides and glycosides while retaining their efficacy in dispelling cold. Refluxing and extracting herbs such as Trichosanthes kirilowii and loquat leaves with a 60% to 80% ethanol solution fully dissolves fat-soluble ingredients such as volatile oils and organic acids, enhancing their cough-relieving and expectorant effects. Percolating herbs such as Gardenia jasminoides and Pinellia ternata with a 45% to 60% ethanol solution not only extracts effective ingredients such as alkaloids and glycosides, but also reduces the dissolution of impurities such as mucus. The above-mentioned targeted extraction strategy designed by the present invention ensures that the effective ingredients of each medicinal material are retained to the maximum extent, laying the foundation for exerting comprehensive medicinal effects.

[0028] The present invention adopts a process of mixed concentration after step-by-step treatment, so that the components obtained by different extraction processes can work together to give full play to the advantages of compound compatibility. The yellow wine components in the steamed product can promote the penetration and absorption of the components in other extraction processes; the polysaccharides and glycosides in the decoction are combined with the fat-soluble components in the alcohol extract to enhance the antiviral and anti-inflammatory effects; the alkaloids and other components in the percolation liquid work together with other components to further enhance the anti-Mycoplasma pneumoniae and antitussive and expectorant effects. The components extracted in each step are fully integrated during the concentration process to form a multi-component, multi-target synergistic system. Compared with a single extraction method, the comprehensive therapeutic effect of the composition in relieving throat and cough, antiviral and anti-Mycoplasma pneumoniae is significantly enhanced.

[0029] In the present invention, after the pulverization process, it is preferably passed through a 20-40 mesh sieve. The present invention does not particularly limit the specific method of pulverization, and conventional pulverization methods in the art can be used. In the present invention, the alcohol content of the rice wine is preferably 14%-18%, and the weight ratio of mixture 1 to rice wine is preferably 10:2-3; the steaming time is preferably 2h-2.5h, more preferably 2.2h-2.3h, and the steaming time is preferably 40min-50min, more preferably 42min-46min.

[0030] In the present invention, the weight ratio of the mixture 2 to water is preferably 1:7-10, more preferably 1:8-9; the soaking time is preferably 20 min-30 min, more preferably 23 min-27 min; the decocting time is preferably 1 h-1.5 h, more preferably 1.2 h-1.4 h.

[0031] In the present invention, when the mixture 3 is refluxed and extracted with an ethanol solution, the volume fraction of the ethanol solution is preferably 65% ​​to 75%, more preferably 70%; the weight ratio of the mixture 3 to the ethanol solution with a volume fraction of 60% to 80% is preferably 1:6 to 8, more preferably 1:7; the number of reflux extractions is preferably 1 to 2 times, and the time for each reflux extraction is preferably 0.5h to 1h, more preferably 0.7h to 0.9h.

[0032] In the present invention, the diafiltration solvent is preferably an ethanol solution with a volume fraction of 50% to 55%, more preferably an ethanol solution with a volume fraction of 52% to 54%; during wetting, the weight ratio of the mixture 4 to the diafiltration solvent is preferably 1:2 to 3, and the wetting time is preferably 1h to 2h, more preferably 1.2h to 1.8h; during impregnation, the weight of the added diafiltration solvent is preferably 4 to 6 times the weight of the mixture 4, more preferably 5 times the weight of the mixture 4, and the impregnation time is preferably 8h to 10h, more preferably 9h; the diafiltration speed is preferably 3 to 5mL per minute, more preferably 4mL per minute.

[0033] The present invention has no particular limitation on the concentration method, and conventional concentration methods in the art may be used. In the present invention, the relative density refers to the relative density at 60°C.

[0034] The preparation method provided by the present invention effectively ensures the quality uniformity and stability of the composition by controlling key process parameters. The steaming time is set to 30 to 60 minutes, which not only ensures the full dissolution of the medicinal ingredients but also avoids decomposition of the ingredients due to excessive steaming. Parameters such as the soaking and boiling time during the water extraction process, the ethanol concentration and reflux time during the alcohol extraction process, and the solvent concentration and collection volume during the filtration process are all scientifically designed to reduce the impact of process fluctuations on the ingredient content. Furthermore, the step-by-step processing avoids mutual interference between medicinal materials of different properties during the same extraction process (for example, the mucilage of jujube may affect the dissolution of other medicinal ingredients), effectively improving the stability of the composition and providing reliable quality assurance for industrial production.

[0035] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0036] In the following examples, unless otherwise specified, all methods are conventional.

[0037] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0038] The quantitative data in the following examples are expressed as average values.

[0039] Example 1 A composition with both soothing and antitussive effects and antiviral effects is prepared from the following raw materials in parts by weight: 11 parts of light fermented black beans, 22 parts of Shenqu, 1 part of Belamcanda chinensis, 2 parts of Platycodon grandiflorum, 5 parts of Perilla frutescens leaves, 32 parts of Citrus aurantium, 1 part of Coptis chinensis, 4 parts of Trichosanthes kirilowii, 5 parts of Loquat leaves, 2 parts of Scutellaria baicalensis, 1 part of Gardenia jasminoides, 5 parts of Magnolia officinalis, 4 parts of Bupleurum chinense, 12 parts of Pinellia ternata, 5 parts of Curcuma aromatica, 12 parts of Licorice root, 4 parts of ginger, 32 parts of jujube, and 4 parts of Poria cocos.

[0040] The preparation method is: Mix coptis root, scutellaria root, magnolia bark and ginger to obtain mixture 1, crush the mixture 1 and pass it through a 20-mesh sieve, take the sieve residue and mix it with yellow rice wine with an alcohol content of 14% and let it steep for 2 hours, the weight ratio of mixture 1 to yellow rice wine being 10:2, and steam the steeped medicine for 30 minutes to obtain a steamed product; Mix light fermented black beans, Shenqu, Belamcanda chinensis, Platycodon grandiflorum, Perilla frutescens leaves, and Citrus aurantium to obtain mixture 2, mix mixture 2 with water, and soak for 20 minutes, with the weight ratio of mixture 2 to water being 1:7, and boil over high heat for 1 hour to obtain the decoction; Mix Trichosanthes kirilowii, loquat leaves, bupleurum and Poria cocos, grind them and pass them through a 20-mesh sieve to obtain a mixture 3, mix the mixture 3 with a 60% by volume ethanol solution, and the weight ratio of the mixture 3 to the 60% by volume ethanol solution is 1:6. Perform reflux extraction twice, the first reflux extraction time is 0.5 h, and the second reflux extraction time is 0.5 h, and collect the extract; Gardenia, Pinellia ternata, Curcuma aromatica, Licorice root and jujube are mixed and crushed through a 20-mesh sieve to obtain a mixture 4. The mixture 4 is mixed and moistened with a percolation solvent for 1 hour, and the weight ratio of the mixture 4 to the percolation solvent is 1:2. After the wetting is completed, absorbent cotton is spread on the bottom of the percolation cylinder to prevent the powder from leaking out. The moistened powder is loaded into the percolation cylinder and compacted. A percolation solvent 4 times the weight of the mixture 4 is added from the top of the percolation cylinder. At the same time, the bottom outlet is opened to exhaust the air in the cylinder, and the outlet is closed. The mixture 4 is immersed in the percolation solvent for 8 hours. After the immersion is completed, the outlet is opened and percolation is started. The percolation speed is controlled to 3 mL per minute, and the percolation liquid is collected to a volume twice the amount of the mixture 4. The percolation solvent is an ethanol solution with a volume fraction of 45%; The decoction, the extract and the percolate are mixed, and concentrated under reduced pressure (the vacuum degree of the reduced pressure concentration is -0.07 MPa and the temperature is 40°C) to a relative density of 1.1 (measured at 60°C) to obtain a concentrate; the concentrate is mixed with the steamed product to obtain the composition.

[0041] Example 2 A composition with both soothing and antitussive effects and antiviral effects is prepared from the following raw materials in parts by weight: 20 parts of light fermented black beans, 26 parts of Shenqu, 3 parts of Belamcanda chinensis, 4 parts of Platycodon grandiflorum, 8 parts of Perilla frutescens leaves, 35 parts of Citrus aurantium, 3 parts of Coptis chinensis, 8 parts of Trichosanthes kirilowii, 9 parts of Loquat leaves, 4 parts of Scutellaria baicalensis, 2 parts of Gardenia jasminoides, 9 parts of Magnolia officinalis, 8 parts of Bupleurum chinense, 16 parts of Pinellia ternata, 8 parts of Curcuma aromatica, 18 parts of Licorice root, 6 parts of ginger, 36 parts of jujube, and 8 parts of Poria cocos.

[0042] The preparation method is: Mix coptis root, scutellaria root, magnolia bark and ginger to obtain mixture 1, crush the mixture 1 and pass it through a 40-mesh sieve, take the sieve residue and mix it with yellow rice wine with an alcohol content of 18% and steep it for 2.5 hours, the weight ratio of mixture 1 to yellow rice wine being 10:3, and steam the steeped medicine for 60 minutes to obtain a steamed product; Mix light fermented black beans, Shenqu, Belamcanda chinensis, Platycodon grandiflorum, Perilla frutescens leaves, and Citrus aurantium to obtain a mixture 2, mix the mixture 2 with water, and soak for 30 minutes, with the weight ratio of the mixture 2 to water being 1:10, and boil over high heat for 1.5 hours to obtain the decoction; Mix Trichosanthes kirilowii, loquat leaves, bupleurum and Poria cocos, grind them and pass them through a 40-mesh sieve to obtain a mixture 3, mix the mixture 3 with an 80% by volume ethanol solution, and the weight ratio of the mixture 3 to the 80% by volume ethanol solution is 1:8, perform reflux extraction once, and collect the extract for 1 hour; Gardenia, Pinellia ternata, Curcuma aromatica, Licorice root and jujube are mixed and crushed through a 40-mesh sieve to obtain a mixture 4. The mixture 4 is mixed and moistened with a percolation solvent for 2 hours, and the weight ratio of the mixture 4 to the percolation solvent is 1:3; after the wetting is completed, absorbent cotton is spread on the bottom of the percolation cylinder to prevent the powder from leaking out, the moistened powder is loaded into the percolation cylinder and compacted, and a percolation solvent 6 times the weight of the mixture 4 is added from the top of the percolation cylinder. At the same time, the bottom outlet is opened to exhaust the air in the cylinder, and the outlet is closed. The mixture 4 is immersed in the percolation solvent for 10 hours. After the immersion is completed, the outlet is opened and percolation is started. The percolation speed is controlled to 5 mL per minute, and the percolation liquid is collected to a volume 4 times the amount of the mixture 4; the percolation solvent is an ethanol solution with a volume fraction of 60%; The decoction, the extract and the percolate are mixed, and concentrated under reduced pressure (the vacuum degree of the reduced pressure concentration is -0.07 MPa and the temperature is 40°C) to a relative density of 1.2 (measured at 60°C) to obtain a concentrate; the concentrate is mixed with the steamed product to obtain the composition.

[0043] Example 3 A composition with both soothing and antitussive effects and antiviral properties is prepared from the following raw materials in parts by weight: 16 parts of light fermented black beans, 24 parts of Shenqu, 2 parts of Belamcanda chinensis, 3 parts of Platycodon grandiflorum, 7 parts of Perilla frutescens leaves, 33 parts of Citrus aurantium, 2 parts of Coptis chinensis, 6 parts of Trichosanthes kirilowii, 7 parts of Loquat leaves, 3 parts of Scutellaria baicalensis, 1 part of Gardenia jasminoides, 7 parts of Magnolia officinalis, 6 parts of Bupleurum chinense, 14 parts of Pinellia ternata, 7 parts of Curcuma aromatica, 15 parts of Licorice root, 5 parts of ginger, 34 parts of jujube, and 6 parts of Poria cocos.

[0044] The preparation method is: Mix coptis root, scutellaria root, magnolia bark and ginger to obtain mixture 1, crush the mixture 1 and pass it through a 30-mesh sieve, take the sieve residue and mix it with yellow rice wine with an alcohol content of 16% and steep it for 2.2 hours, the weight ratio of mixture 1 to yellow rice wine being 10:3, and steam the steeped medicine for 44 minutes to obtain a steamed product; Mix light fermented black beans, Shenqu, Belamcanda chinensis, Platycodon grandiflorum, Perilla frutescens leaves, and Citrus aurantium to obtain mixture 2, mix mixture 2 with water, and soak for 25 minutes, with the weight ratio of mixture 2 to water being 1:8, and boil over high heat for 1.3 hours to obtain the decoction; Mix Trichosanthes kirilowii, loquat leaves, bupleurum and Poria cocos, grind them and pass them through a 30-mesh sieve to obtain a mixture 3, mix the mixture 3 with a 70% by volume ethanol solution, and the weight ratio of the mixture 3 to the 70% by volume ethanol solution is 1:7. Perform reflux extraction twice, the first reflux extraction time is 0.8 h, and the second reflux extraction time is 0.8 h, and collect the extract; Gardenia, Pinellia ternata, Curcuma aromatica, Licorice root and jujube are mixed and crushed through a 30-mesh sieve to obtain a mixture 4. The mixture 4 is mixed and moistened with a percolation solvent for 1.5 hours, and the weight ratio of the mixture 4 to the percolation solvent is 1:2. After the wetting is completed, absorbent cotton is spread on the bottom of the percolation cylinder to prevent the powder from leaking out. The moistened powder is loaded into the percolation cylinder and compacted. A percolation solvent 5 times the weight of the mixture 4 is added from the top of the percolation cylinder. At the same time, the bottom outlet is opened to exhaust the air in the cylinder, and the outlet is closed. The mixture 4 is immersed in the percolation solvent for 9 hours. After the immersion is completed, the outlet is opened and percolation is started. The percolation speed is controlled to 4 mL per minute, and the percolation liquid is collected to a volume 3 times the amount of the mixture 4. The percolation solvent is a 50% by volume ethanol solution. The decoction, the extract and the percolate are mixed, and concentrated under reduced pressure (the vacuum degree of the reduced pressure concentration is -0.07 MPa and the temperature is 40°C) to a relative density of 1.1 (measured at 60°C) to obtain a concentrate; the concentrate is mixed with the steamed product to obtain the composition.

[0045] Comparative Example 1 A throat-clearing and cough-relieving granule is prepared by the following steps: Step 1: Take 15 parts of bupleurum, 9 parts of scutellaria, 9 parts of pinellia, 6 parts of belamcanda, 10 parts of platycodon, 6 parts of gardenia, 6 parts of light fermented black beans, 15 parts of aurantium, 12 parts of loquat leaves, 15 parts of trichosanthes, 3 parts of coptis, 10 parts of shenqu, 15 parts of perilla leaves, 15 parts of magnolia bark, 15 parts of poria, 20 parts of turmeric, 10 parts of liquorice, 15 parts of ginger, and 10 parts of jujube, a total of 19 medicinal materials, and pass them through a 10-mesh sieve respectively.

[0046] Step 2: Take the medicinal materials sieved in step 1, and except Shenqu and Poria, add 10 times the amount of water to the remaining 17 medicinal materials, heat and reflux and extract once, and the extraction time is 1.5 hours.

[0047] Step 3: Add Shenqu and Poria to the extract of step 2, and then reflux and extract once for 1.5 hours.

[0048] Step 4: Filter the extract, combine the filtrates and concentrate them into an extract (relative density 1.25).

[0049] Step 5: Wet granulation: Add auxiliary materials in a ratio of 1:2, wherein the auxiliary materials are an appropriate amount of low-viscosity dextrin, and make granules and dry them at a drying temperature of 60°C.

[0050] Comparative Example 2 The difference from Example 3 is that the amount of light fermented black beans is 25 parts, the amount of Shenqu is 30 parts, and the rest are the same as Example 3.

[0051] Comparative Example 3 The difference from Example 3 is that mixture 1 is not steamed with rice wine, but is directly decocted with water. Specifically, mixture 1 and mixture 2 are mixed to obtain a composite mixture; then 8 times the weight of water of the composite mixture is added and soaked for 25 minutes. After the soaking is completed, the mixture is decocted over high heat for 1.3 hours to obtain the decoction; the rest is the same as in Example 3.

[0052] Comparative Example 4 The difference from Example 3 is that the volume fraction of the ethanol solution used for reflux extraction of mixture 3 is 50%, the diafiltration solvent is an ethanol solution with a volume fraction of 70%, and the rest is the same as Example 3.

[0053] Example 4 Verification of throat-relieving and cough-relieving effects Healthy adult mice (weighing 18-22 g) were selected and maintained in an experimental animal housing environment at a temperature of (22 ± 2)°C, a humidity of 50%-60%, and a 12-hour day and night light cycle. The animals were acclimated for 5 days before the experiment. The animals were randomly divided into 9 groups, 10 mice each, including a blank control group, a positive drug group, Example 1 group, Example 2 group, Example 3 group, Comparative Example 1 group, Comparative Example 2 group, Comparative Example 3 group, and Comparative Example 4 group.

[0054] All groups of mice were placed in a sealed glass container and sprayed with 25% ammonia water at a constant rate of 2 mL / min using an ultrasonic nebulizer for 30 seconds. After the spray was completed, a stopwatch was immediately started and the number of coughs within 5 minutes was recorded. Three consecutive abdominal muscle contractions accompanied by a typical puffing sound were considered an effective cough. If the number of coughs reached 10 or more within 5 minutes, the ammonia cough model was successfully established.

[0055] After the modeling was successful, mice in each group were taken out and gavage-administered the corresponding group of drugs, wherein the blank control group was gavage-administered with normal saline at a dose of 15 mg / kg; the positive drug group was gavage-administered with dextromethorphan at a dose of 15 mg / kg; the Example 1 group was gavage-administered with the composition obtained in Example 1 at a dose of 10 mg / kg; the Example 2 group was gavage-administered with the composition obtained in Example 2 at a dose of 10 mg / kg; the Example 3 group was gavage-administered with the composition obtained in Example 3 at a dose of 10 mg / kg; the comparative example 1 group was gavage-administered with the composition obtained in comparative example 1 at a dose of 10 mg / kg; the comparative example 2 group was gavage-administered with the composition obtained in comparative example 2 at a dose of 10 mg / kg; the comparative example 3 group was gavage-administered with the composition obtained in comparative example 3 at a dose of 10 mg / kg; the comparative example 4 group was gavage-administered with the composition obtained in comparative example 4 at a dose of 10 mg / kg. Each group was gavage-treated once daily for three consecutive days. One hour after the last dose, coughing was induced again using the same method used for modeling. The time from exposure to ammonia vapor to the first cough (cough latency) and the number of coughs within 5 minutes were recorded. Cough suppression rate was calculated as follows: Cough suppression rate (%) = [(average number of coughs in the blank control group - average number of coughs in the experimental group) / average number of coughs in the blank control group] × 100%. Finally, blood was collected from each mouse, centrifuged, and serum was separated for assay of cytokine IL-6 content. The results are shown in Table 1. This demonstrates that the composition of the present invention has significant throat-clearing and cough-relieving effects and can reduce the levels of inflammatory factors.

[0056] Table 1 Verification results of the throat-relieving and cough-relieving effects

[0057] Example 5 Verification of anti-respiratory syncytial virus effect 1. Test preparation: Cell line: Human airway epithelial cell line BEAS-2B (ATCC CRL-9609) was used. After thawing, cells were cultured in DMEM (Gibco) supplemented with 10% fetal bovine serum (Gibco) and 1% double-streptomycin (100 U / mL penicillin and 100 μg / mL streptomycin, Gibco) at 37°C in a 5% CO2 incubator. When cells reached the logarithmic growth phase (80%-90% confluency), they were digested and passaged using 0.25% trypsin-EDTA (Gibco) for subsequent experiments.

[0058] Virus strain: Respiratory syncytial virus A2 (RSV A2) was selected. After virus recovery, it was amplified in Vero cells. Before use, the endpoint dilution method (TCID 50 ) Determine the virus titer: dilute the virus 10-fold (10 -1 ~10 -10), were inoculated into Vero cells in 96-well plates (100 μL per well, 8 replicates per dilution), and after 72 hours of culture, the cytopathic effect was observed and the TCID was calculated using the Reed-Muench method. 50 Ensure that the virus concentration used in each experiment is 1×10 6 TCID 50 / mL.

[0059] Preliminary toxicity test of the compositions obtained in Examples 1 to 3 and the compositions obtained in Comparative Examples 1 to 4 on the human airway epithelial cell line BEAS-2B: The composition obtained in Example 1 was diluted into solutions with different concentrations (100%, 50%, 25%, 12.5%, 6.25%, and 3.125%) using culture medium. The compositions obtained in Examples 2-3 and Comparative Examples 1-4 were diluted into the above six concentration gradient solutions according to the above steps. A blank control group (containing only cells and culture medium) was also set up. BEAS-2B cells in the logarithmic growth phase were trypsinized and the cell concentration was adjusted to 5×10 4 cells / mL, seeded into 96-well plates, 100 μL per well, and cultured in a 37°C, 5% CO2 incubator for 24 h to allow the cells to adhere to the plate and grow to 80%-90% confluency.

[0060] Dosing treatment: the original culture medium in the 96-well plate was discarded, each well was washed twice with PBS, and 100 μL of the corresponding concentration of the composition solution was added to each composition concentration group, and 100 μL of normal culture medium was added to the blank control group.

[0061] Culture and Detection: The 96-well plate was placed in a 37°C, 5% CO2 incubator for continued incubation. At 24, 48, and 72 hours, 10 μL of CCK-8 reagent was added to each well. After an additional 2 hours of incubation, the absorbance at 450 nm (OD value) was measured using a microplate reader. Cell viability was calculated as follows: Cell viability (%) = (OD value of experimental group / OD value of blank control group) × 100%.

[0062] The results showed that the cell viability of each composition group at various time points (24, 48, and 72 hours) was ≥80% across multiple concentration gradients, and the cells exhibited normal morphology (observed under an inverted microscope, the cells were polygonal or spindle-shaped, adherent and well-grown, with no obvious signs of cell rounding, shrinkage, or shedding). This indicates that the compositions had no significant toxicity to BEAS-2B cells within these concentration ranges. Therefore, they were added directly to the antiviral cell experiments described below.

[0063] 2. Medication and viral infection Cell inoculation: BEAS-2B cells in the logarithmic growth phase were trypsinized and the cell concentration was adjusted to 5×10 4 cells / mL, seeded into 96-well plates (100 μL per well), and cultured in a 37°C, 5% CO2 incubator for 24 hours until the cells adhered to the wall and grew to 80%-90% confluency.

[0064] Virus infection and drug administration: Aspirate the original culture medium in the 96-well plate and wash each well twice with PBS, 100 μL each time (96-well plate). Gently shake the culture plate to remove residual serum.

[0065] The trial groups were: Blank control group: only BEAS-2B cells and normal culture medium were added without any treatment.

[0066] Virus control group: RSV A2 strain was inoculated (MOI=1) without adding any drugs.

[0067] Positive drug control group: After inoculation with RSV A2 strain, ribavirin at a concentration of 10 μg / mL was added.

[0068] Group 1 of Example 1: After inoculation with RSV A2 strain, the sterilized composition obtained in Example 1 was added at a concentration of 5 μg / mL.

[0069] Group 2 of Example 2: After inoculation with RSV A2 strain, the sterilized composition obtained in Example 2 was added at a concentration of 5 μg / mL.

[0070] Group 3 of Example: After inoculation with RSV A2 strain, the sterilized composition obtained in Example 3 was added at a concentration of 5 μg / mL.

[0071] Comparative Example 1 group: After inoculation with RSV A2 strain, the sterilized composition obtained in Comparative Example 1 was added at a concentration of 5 μg / mL.

[0072] Comparative Example 2 group: After inoculation with RSV A2 strain, the sterilized composition obtained in Comparative Example 2 at a concentration of 5 μg / mL was added.

[0073] Comparative Example 3 group: After inoculation with RSV A2 strain, the sterilized composition obtained in Comparative Example 3 at a concentration of 5 μg / mL was added.

[0074] Comparative Example 4 group: After inoculation with RSV A2 strain, the sterilized composition obtained in Comparative Example 4 was added at a concentration of 5 μg / mL.

[0075] 100 μL of serum-free DMEM medium containing RSV A2 strain (virus concentration of 1×10 6TCID 50 / mL, MOI=1), 100 μL serum-free DMEM medium was added to the blank control group and incubated in a 37°C, 5% CO2 incubator for 2 hours. During this period, the culture plate was gently shaken every 30 minutes to allow the virus to fully adsorb on the cell surface.

[0076] After the incubation, the virus solution was discarded, and the culture medium containing the corresponding drug was added to each well of each drug treatment group (100 μL / well). The blank control group and the virus control group were added with normal culture medium, and the culture was continued for 48 hours (preliminary experiments determined that the virus-induced cytopathic effect was significant during this time period).

[0077] After the incubation period, 10 μL of CCK-8 reagent was added to each well. The culture plate was gently shaken to evenly distribute the reagent, and the cells were incubated for another 2 hours in a 37°C, 5% CO2 incubator. The absorbance (OD) at 450 nm was measured using a microplate reader, and cell viability was calculated as follows: cell viability (%) = (OD value of experimental group / OD value of blank control group) × 100%. This was used to evaluate the protective effect of the traditional Chinese medicine composition on cell viability after viral infection. The results are shown in Table 2.

[0078] Table 2 Verification results of anti-respiratory syncytial virus effect

[0079] Example 6 Verification of anti-influenza A virus strain A / PR / 8 / 34 / (H1N1) activity 1. Test preparation: Cell line: MDCK cells (ATCC CCL-34) were revived and cultured in MEM medium supplemented with 10% fetal bovine serum and 1% double-antibody (penicillin 100 U / mL, streptomycin 100 μg / mL) in a 37°C, 5% CO2 incubator. When the cells reached the logarithmic growth phase (cell confluence reached 80%-90%), they were digested with 0.25% trypsin-EDTA and passaged for subsequent experiments.

[0080] Virus strain: Influenza A virus A / PR / 8 / 34 / (H1N1) strain was amplified in SPF chicken embryos: the virus was inoculated into the allantoic cavity of 10-11 day old chicken embryos, incubated at 35℃ for 48-72 hours, and then the allantoic fluid was collected. 50 The virus titer was determined by PCR and the concentration was adjusted to 1×10 6 TCID 50 / mL to ensure consistent virus concentration in the experiments.

[0081] Preliminary toxicity test on MDCK cells of the compositions obtained in Examples 1 to 3 and the compositions obtained in Comparative Examples 1 to 4: MDCK cells were cultured at 5×104 Cells were seeded in 96-well plates at a density of 100 μL per well at 100 cells / mL and cultured for 24 hours until 80% confluency was achieved. The original culture medium was aspirated and the experimental groups were treated with culture medium containing different concentrations of the traditional Chinese medicine composition (100 μL / well). The culture medium for these different concentrations of the traditional Chinese medicine composition was prepared by using MEM medium to prepare a series of concentrations of 100, 50, 25, 12.5, 6.25, and 3.125 mg / mL, followed by sterilization by filtration through a 0.22 μm filter. A blank control group was treated with normal culture medium. Six replicate wells were set up for each group.

[0082] After culturing for 24, 48, and 72 hours, 10 μL of CCK-8 reagent was added to each well. The absorbance at 450 nm was measured after 2 hours of incubation, and the cell viability was calculated: cell viability (%) = (OD value of experimental group / OD value of blank control group) × 100%.

[0083] The results showed that the cell viability in each concentration group was ≥80%, with no concentration-dependent decrease. Therefore, it was directly added in the antiviral cell experiment described below.

[0084] 2. Medication and viral infection Cell inoculation: Digest the MDCK cells in the logarithmic growth phase and adjust the concentration to 5×10 4 cells / mL, seeded in 96-well plates (100 μL per well) for CCK-8 assays; seeded in 96-well plates (500 μL per well) for other assays. Incubate at 37°C, 5% CO₂ for 24 hours until cells adhere to the plate and reach 80%–90% confluency.

[0085] Virus infection and drug administration: Aspirate and discard the culture medium in the wells, wash twice with PBS to remove residual serum.

[0086] Experimental groups: Blank control group: Contains only MDCK cells and normal culture medium, without virus infection and drug treatment.

[0087] Virus control group: Inoculated with influenza A virus A / PR / 8 / 34 / (H1N1) strain (MOI=1) without adding any drugs.

[0088] Positive drug control group: After inoculation with H1N1 virus, oseltamivir at a concentration of 1 μM was added.

[0089] Group 1 of Example: After inoculation with H1N1 virus, the sterilized composition obtained in Example 1 was added at a concentration of 1 μg / mL.

[0090] Group 2 of Example: After inoculation with the H1N1 virus, the sterilized composition obtained in Example 2 was added at a concentration of 1 μg / mL.

[0091] Group 3 of Example: After inoculation with H1N1 virus, the sterilized composition obtained in Example 3 was added at a concentration of 1 μg / mL.

[0092] Comparative Example 1 group: After inoculation with H1N1 virus, the sterilized composition obtained in Comparative Example 1 was added at a concentration of 1 μg / mL.

[0093] Comparative Example 2 group: After inoculation with H1N1 virus, the sterilized composition obtained in Comparative Example 2 was added at a concentration of 1 μg / mL.

[0094] Comparative Example 3 group: After inoculation with H1N1 virus, the sterilized composition obtained in Comparative Example 3 was added at a concentration of 1 μg / mL.

[0095] Comparative Example 4 group: After inoculation with H1N1 virus, the sterilized composition obtained in Comparative Example 4 was added at a concentration of 1 μg / mL.

[0096] 100 μL of serum-free MEM medium containing virus was added to each well of the virus control group and each drug-treated group (MOI=1), and serum-free MEM medium was added to the blank control group. The cells were incubated at 37°C and 5% CO2 for 1.5 hours. During this period, the culture plate was gently shaken every 30 minutes to promote virus adsorption.

[0097] After the incubation, the virus solution was discarded, and the culture medium containing the corresponding drug (100 μL / well) was added to each drug treatment group, and normal culture medium was added to the blank control group and the virus control group. The culture was continued for 24 hours (preliminary experiments determined that the virus-induced cytopathic effect was significant during this time period).

[0098] After the incubation period, 10 μL of CCK-8 reagent was added to each well of the 96-well plate and incubated at 37°C for 2 hours. The OD value at 450 nm was measured with a microplate reader, and the cell viability was calculated as follows: cell viability (%) = (OD value of experimental group / OD value of blank control group) × 100%. The results are shown in Table 3.

[0099] Table 3 Results of validation of anti-influenza A virus strain A / PR / 8 / 34 / (H1N1)

[0100] Example 7 Verification of anti-Mycoplasma pneumoniae effect 1. Test preparation: A549 cell culture: After thawing, A549 cells (ATCC CCL-185) were placed in F12K medium containing 10% fetal bovine serum and 1% double-antibody (penicillin 100 U / mL, streptomycin 100 μg / mL) and cultured in a 37°C, 5% CO2 incubator. When the cells were in the logarithmic growth phase and the confluence reached 80%-90%, they were digested and passaged with 0.25% trypsin-EDTA and the cell density was adjusted to 5×10 4 cells / mL for future use.

[0101] Culture of Mycoplasma pneumoniae ATCC 29342: Inoculate the strain into SP4 liquid medium (containing 10% horse serum and 0.5% yeast extract) and incubate at 37°C. Observe for color changes (phenol red indicator) every 2–3 days. Verify the purity of the culture using PCR. Adjust the bacterial concentration by serial dilution before use. The optimal multiplicity of infection (MOI = 10) is determined through preliminary experiments.

[0102] Preliminary toxicity test of the compositions obtained in Examples 1 to 3 and the compositions obtained in Comparative Examples 1 to 4 on A549 cells: A series of concentration gradient solutions of each composition were prepared using F12K medium: 1000 μg / mL, 500 μg / mL, 250 μg / mL, 125 μg / mL, 62.5 μg / mL, and 31.25 μg / mL. The solutions were sterilized by filtration with a 0.22 μm filter membrane and set aside.

[0103] A549 cells in the logarithmic growth phase were digested with 0.25% trypsin-EDTA and the concentration was adjusted to 5×10 4 Cells were seeded in 96-well plates at 100 μL per well at 37°C, 5% CO₂ incubator. After cells adhered for 24 hours, the culture medium was aspirated and the cells were washed twice with PBS. Each experimental group received the corresponding treatment solution, while the blank control group received normal culture medium and continued to be cultured at 37°C, 5% CO₂. At 24, 48, and 72 hours, 10 μL of CCK-8 reagent was added to each well. After an additional 2 hours of incubation, the absorbance at 450 nm (OD) was measured using a microplate reader. Cell viability was calculated as follows: cell viability (%) = (OD value of experimental group / OD value of blank control group) × 100%. Results showed that cell viability was ≥80% in all concentration groups, with no concentration-dependent decrease. Therefore, the cells were added directly in the following anti-Mycoplasma pneumoniae cell assay.

[0104] 2. Medication and infection Cell seeding: A549 cells of adjusted concentration were seeded into a 96-well plate (100 μL per well for CCK-8 assay) and cultured at 37°C, 5% CO2 for 24 hours until the cells adhered to the plate and reached 80%-90% confluency.

[0105] Infection and drug administration: Aspirate and discard the culture medium in the wells, and wash twice with PBS.

[0106] Experimental groups: Blank control group: only containing A549 cells and normal F12K culture medium.

[0107] Mycoplasma pneumoniae infection control group: inoculated with Mycoplasma pneumoniae ATCC 29342 (MOI=10), without adding drugs.

[0108] Positive drug control group: After inoculation with Mycoplasma pneumoniae ATCC 29342 (MOI = 10), azithromycin with 1 μM of virus was added.

[0109] Group 1 of Example: After inoculation with Mycoplasma pneumoniae ATCC 29342 (MOI=10), the sterilized composition obtained in Example 1 was added at a concentration of 0.5 μg / mL.

[0110] Group 2 of Example 2: After inoculation with Mycoplasma pneumoniae ATCC 29342 (MOI=10), the sterilized composition obtained in Example 2 was added at a concentration of 0.5 μg / mL.

[0111] Group 3 of Example: After inoculation with Mycoplasma pneumoniae ATCC 29342 (MOI=10), the sterilized composition obtained in Example 3 was added at a concentration of 0.5 μg / mL.

[0112] Comparative Example 1 group: After inoculating Mycoplasma pneumoniae ATCC 29342 (MOI=10), the sterilized composition obtained in Comparative Example 1 was added at a concentration of 0.5 μg / mL.

[0113] Comparative Example 2 group: After inoculating Mycoplasma pneumoniae ATCC 29342 (MOI=10), the sterilized composition obtained in Comparative Example 2 was added at a concentration of 0.5 μg / mL.

[0114] Comparative Example 3 group: After inoculating Mycoplasma pneumoniae ATCC 29342 (MOI=10), the sterilized composition obtained in Comparative Example 3 was added at a concentration of 0.5 μg / mL.

[0115] Comparative Example 4 group: After inoculating Mycoplasma pneumoniae ATCC 29342 (MOI=10), the sterilized composition obtained in Comparative Example 4 was added at a concentration of 0.5 μg / mL.

[0116] 100 μL of culture medium containing M. pneumoniae (MOI = 10) was added to each well of the M. pneumoniae control group and each drug-treated group. Normal culture medium was added to the blank control group and incubated at 37°C, 5% CO₂ for 48 hours. After incubation, 100 μL of culture medium containing the corresponding drug was added to each well of each drug-treated group. Normal culture medium was added to the blank control group and the M. pneumoniae control group, and culture continued for 48 hours. After the incubation period, 10 μL of CCK-8 reagent was added to each well of the 96-well plate. The plates were incubated at 37°C for 2 hours. The OD value at 450 nm was measured with a microplate reader, and cell viability was calculated as follows: Cell viability (%) = (OD value of experimental group / OD value of blank control group) × 100%. The results are shown in Table 4.

[0117] Table 4 Verification results of anti-Mycoplasma pneumoniae effect

[0118] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A composition having both throat-relieving and cough-relieving and antiviral effects, characterized in that: The invention comprises the following raw materials in parts by weight: 11-20 parts of light fermented black beans, 22-26 parts of Shenqu, 1-3 parts of Belamcanda chinensis, 2-4 parts of Platycodon grandiflorum, 5-8 parts of Perilla frutescens leaves, 32-35 parts of Citrus aurantium, 1-3 parts of Coptis chinensis, 4-8 parts of Trichosanthes kirilowii, 5-9 parts of Loquat leaves, 2-4 parts of Scutellaria baicalensis, 1-2 parts of Gardenia jasminoides, 5-9 parts of Magnolia officinalis, 4-8 parts of Bupleurum chinense, 12-16 parts of Pinellia ternata, 5-8 parts of Curcuma aromatica, 12-18 parts of Licorice root, 4-6 parts of Ginger, 32-36 parts of Jujube and 4-8 parts of Poria cocos.

2. The composition according to claim 1, characterized in that The antiviral agent includes anti-respiratory syncytial virus or anti-influenza virus.

3. The method for preparing the composition according to claim 1 or 2, characterized in that: The steps include: Mixing coptis root, scutellaria root, magnolia bark and ginger to obtain mixture 1, crushing the mixture 1 and mixing it with rice wine to moisten it, and steaming the moistened medicine for 30 minutes to 60 minutes to obtain a steamed product; Mix light fermented black beans, Shenqu, Belamcanda chinensis, Platycodon grandiflorum, Perilla frutescens leaves and Citrus aurantium to obtain mixture 2, mix mixture 2 with water, soak, and boil to obtain a decoction; Mix and grind Trichosanthes kirilowii, loquat leaves, bupleurum and Poria cocos to obtain a mixture 3, mix the mixture 3 with a 60% to 80% by volume ethanol solution, perform reflux extraction, and collect the extract; Gardenia jasminoides, pinellia tuber, turmeric, liquorice and jujube are mixed and crushed to obtain mixture 4, the mixture 4 is mixed and moistened with a percolation solvent, and then the percolation solvent is added for impregnation. After the impregnation is completed, percolation is started, and the percolation liquid is collected to a volume of 2 to 4 times the amount of mixture 4; the percolation solvent is an ethanol solution with a volume fraction of 45% to 60%; The decoction, the extract and the percolated liquid are mixed and concentrated to a relative density of 1.1 to 1.2 to obtain a concentrate; the concentrate is mixed with the steamed product to obtain the composition.

4. The preparation method according to claim 3, characterized in that After crushing, pass through a 20-40 mesh sieve.

5. The preparation method according to claim 3, characterized in that The weight ratio of the mixture 1 to the rice wine is 10:2-3; and the steeping time is 2h-2.5h.

6. The preparation method according to claim 3, characterized in that The weight ratio of the mixture 2 to water is 1:7-10; the soaking time is 20 min-30 min; and the decoction time is 1 h-1.5 h.

7. The preparation method according to claim 3, characterized in that The weight ratio of the mixture 3 to the ethanol solution with a volume fraction of 60% to 80% is 1:6 to 8; the number of reflux extractions is 1 to 2 times, and the time for each reflux extraction is 0.5 h to 1 h.

8. The preparation method according to claim 3, characterized in that During wetting, the weight ratio of mixture 4 to diafiltration solvent is 1:2~3, and the wetting time is 1h~2h; during impregnation, the weight of the added diafiltration solvent is 4~6 times the weight of mixture 4, and the impregnation time is 8h~10h; the diafiltration speed is 3~5mL per minute.

9. Use of the composition according to claim 1 or 2 or the preparation method according to any one of claims 3 to 8 in the preparation of an anti-Mycoplasma pneumoniae product.

Images