A traditional Chinese medicine composition, preparation, preparation method and application

Abstract

This invention belongs to the field of traditional Chinese medicine (TCM) technology, and particularly relates to a TCM composition, preparation, preparation method, and application. The TCM composition includes areca nut, magnolia bark, cardamom, anemarrhena rhizome, white peony root, scutellaria root, licorice root, honeysuckle flower, forsythia fruit, malt, hawthorn fruit, medicated leaven, and tangerine peel. Experimental results show that after administration of this TCM composition, the IL-10 content in the bronchoalveolar lavage fluid of mice significantly increased; the contents of IL-1β, TNF-α, and IL-6 significantly decreased. This indicates that the TCM composition can reduce lung tissue inflammatory damage by inhibiting the release of pro-inflammatory cytokines IL-1β, TNF-α, and IL-6, and promoting the content of the anti-inflammatory cytokine IL-10. This indicates that the TCM composition has a good therapeutic effect in antiviral inflammatory responses. The drug for antiviral inflammatory responses is a traditional Chinese medicine preparation, safe to use, and the preparation method is simple, rapid, efficient, and easy to operate. The prepared mixtures, granules, pills, tablets, capsules, etc., are easy to carry and convenient to take.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, and in particular to a traditional Chinese medicine composition, preparation, preparation method and application. Background Technology

[0002] Influenza is an acute respiratory infectious disease caused by influenza viruses (mainly classified as type A, B, and C). It is highly contagious and spreads through droplets and contact. Its main symptoms include sudden high fever (usually >38.5℃), muscle aches, fatigue, headache, cough, sore throat, and gastrointestinal symptoms (such as nausea and diarrhea) in some patients. Children, the elderly, patients with chronic diseases, and pregnant women are more prone to severe illness. Viral clearance mainly relies on the body's immune system; however, the inflammatory response triggered by these viruses can severely damage the body's vital functions. Therefore, antiviral treatment of inflammatory responses is of significant clinical importance. Traditional Chinese medicine considers influenza to fall under the category of "external febrile diseases," often caused by wind-heat or wind-cold pathogens invading the lungs and wei qi. Clinical manifestations include fever, cough, headache, and muscle aches. Traditional treatment emphasizes "relieving exterior pathogens, clearing heat and detoxifying, and relieving cough."

[0003] Currently, the main antiviral drugs used in clinical practice include neuraminidase inhibitors (oseltamivir, zanamivir, peramivir) and RNA polymerase inhibitors (baloxavir marboxil, favipiravir). These drugs generally suffer from problems such as easy development of drug resistance and significant side effects. Drugs used to treat antiviral-induced inflammatory responses mainly include nonsteroidal anti-inflammatory drugs (NSAIDs), such as ibuprofen, and glucocorticoids, such as hydrocortisone. While these anti-inflammatory drugs have achieved good clinical efficacy, their proven toxic side effects have brought challenges to clinical treatment.

[0004] Anti-inflammatory traditional Chinese medicines mainly include Lianhua Qingwen capsules and Jinhua Qinggan granules, but they also have problems such as slow onset of action and incomplete symptom relief.

[0005] Traditional Chinese medicine (TCM) compound formulas have a long history and unique advantages in the treatment of influenza, exerting antiviral, immune-regulating, and symptom-relieving effects through multiple components, targets, and pathways. However, current clinical applications of TCM for antiviral infection and inflammatory response still face challenges such as unstable efficacy and unclear mechanisms of action. Summary of the Invention

[0006] In view of this, the technical problem to be solved by the present invention is to provide a traditional Chinese medicine composition, a preparation thereof, a method for preparing the preparation, and its application in the preparation of a drug for antiviral infection and inflammatory response.

[0007] The first aspect of the present invention provides a traditional Chinese medicine composition comprising the following raw materials in parts by weight: 1-20 parts of areca nut, 1-20 parts of magnolia bark, 1-20 parts of cardamom, 1-20 parts of anemarrhena rhizome, 1-20 parts of white peony root, 1-20 parts of scutellaria root, 1-20 parts of licorice root, 1-20 parts of honeysuckle flower, 1-20 parts of forsythia fruit, 1-20 parts of malt, 1-20 parts of hawthorn fruit, 1-20 parts of medicated leaven, and 1-20 parts of dried tangerine peel.

[0008] Preferably, in some embodiments, the mass fractions of each component in the traditional Chinese medicine composition are: 9 parts areca nut, 9 parts magnolia bark, 6 parts cardamom, 9 parts anemarrhena rhizome, 9 parts white peony root, 9 parts scutellaria root, 6 parts licorice root, 9 parts honeysuckle, 9 parts forsythia fruit, 9 parts malt, 9 parts hawthorn, 9 parts medicated leaven, and 6 parts tangerine peel.

[0009] More preferably, in some embodiments, the processing specifications of malt, hawthorn, and Shenqu in the traditional Chinese medicine composition are charred malt, charred hawthorn, and charred Shenqu.

[0010] A second aspect of the present invention provides a traditional Chinese medicine preparation, which is mainly made of the above-mentioned traditional Chinese medicine composition.

[0011] Preferably, in some embodiments, the traditional Chinese medicine preparation is made from a traditional Chinese medicine composition and pharmaceutically acceptable excipients.

[0012] Preferably, the dosage forms of the medicine of the present invention include, but are not limited to, decoctions, granules, capsules, tablets, mixtures, pills, tinctures, syrups or sprays.

[0013] In some embodiments provided by the present invention, the capsule is a hard capsule or a soft capsule.

[0014] In some embodiments provided by the present invention, the tablet is an oral tablet or a dental tablet.

[0015] Oral tablets refer to tablets intended for oral administration. Most of these tablets contain drugs that are absorbed through the gastrointestinal tract to exert their effects, while some contain drugs that exert their effects locally within the gastrointestinal tract. In some embodiments provided by this invention, the oral tablets are ordinary compressed tablets, dispersible tablets, effervescent tablets, chewable tablets, coated tablets, or sustained-release tablets.

[0016] Pharmaceutically acceptable excipients include one or more of the following: fruit powder, flavoring, sweetener, acidulant, filler, lubricant, preservative, suspending agent, food coloring, diluent, emulsifier, disintegrant, or plasticizer.

[0017] The third aspect of this invention provides a method for preparing the traditional Chinese medicine preparation provided in the second aspect, comprising: adding water to areca nut, magnolia bark, cardamom, anemarrhena rhizome, white peony root, scutellaria root, licorice root, honeysuckle flower, forsythia fruit, malt, hawthorn fruit, medicated leaven, and tangerine peel, and then filtering, concentrating, and drying the decoction to obtain the final product.

[0018] In some embodiments, the mass ratio of water to the herbal composition is (4-40):1 during decoction, and the decoction is performed 1-3 times, each time for 1-2 hours.

[0019] Preferably, areca nut, magnolia bark, cardamom, anemarrhena rhizome, white peony root, scutellaria root, licorice root, honeysuckle flower, forsythia fruit, malt, hawthorn fruit, medicated leaven, and tangerine peel are decocted twice with water, the first time with 11 times the amount of water and the second time with 9 times the amount of water, each decoction lasting 1.5 hours. The decoction is soaked for 30 minutes before the first decoction. The decoctions are combined, filtered, and the filtrate is concentrated under reduced pressure to an extract with a relative density of approximately 1.05–1.30 (50℃–60℃), followed by concentration and drying. More preferably, the density of the extract is 1.10.

[0020] More preferably, the drying is carried out by spray drying, which further includes the steps of adding maltodextrin, mixing, and forming granules.

[0021] Acute lung injury (ALI) is a diffuse inflammatory condition affecting the capillary epithelium and alveolar epithelium of the lungs. Clinical symptoms often include chest tightness and shortness of breath. If ALI is not controlled in time, it can develop into acute respiratory distress syndrome (ARDS), characterized by severe hypoxemia and diffuse alveolar damage, with a mortality rate as high as 40%.

[0022] Viral infection is a major cause of acute lung injury. Its pathogenesis is the result of the combined effects of the direct cytotoxic effects of the virus and the excessive immune response of the host ("cytokine storm"), ultimately leading to the disruption of the alveolar-capillary barrier, non-cardiac pulmonary edema, and severe respiratory failure.

[0023] Experiments in the embodiments of this invention show that the traditional Chinese medicine composition can be used to treat inflammatory responses to viral infections.

[0024] Furthermore, the present invention also provides a method for treating inflammatory responses to viral infections, comprising administering the traditional Chinese medicine composition of the present invention.

[0025] The administration methods of the traditional Chinese medicine composition include, but are not limited to, oral administration, spray inhalation, rectal administration, nasal administration, buccal administration, vaginal administration, and topical administration. Alternatively, it can be administered via non-enteric routes, such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal, or intracranial injection or infusion; or via an external implantation device. Preferably, the administration method is oral, intraperitoneal, or intravenous administration.

[0026] In this invention, the traditional Chinese medicine composition and other therapeutic agents may exist independently or in combination; this invention does not limit this.

[0027] In this invention, when the drug includes a traditional Chinese medicine composition and other therapeutic agents, two or more drugs may be administered simultaneously or sequentially, and this invention does not limit this.

[0028] For example, the herbal composition in the drug is applied no longer than 6 hours before the application of other therapeutic agents, preferably no longer than 5 hours, more preferably no longer than 4 hours, even more preferably no longer than 3 hours, more preferably no longer than 2 hours and most preferably no longer than 1 hour and / or no longer than 6 hours after the application of other therapeutic agents, preferably no longer than 5 hours, more preferably no longer than 4 hours, even more preferably no longer than 3 hours, more preferably no longer than 2 hours and most preferably no longer than 1 hour.

[0029] In this invention, when the drug includes a traditional Chinese medicine composition and other therapeutic agents, the dosing frequencies of two or more drugs may be the same or different, and this invention does not limit this.

[0030] For example, the dosing frequency of two or more drugs in a drug is independently selected from 4 times / day, 3 times / day, 2 times / day or 1 time / day, wherein the dosing frequency of the traditional Chinese medicine composition is 4 times / day, 3 times / day, 2 times / day or 1 time / day.

[0031] In this invention, the drug comprises a traditional Chinese medicine composition, and the dosage administered is not less than the effective dose. If the drug comprises a traditional Chinese medicine composition and other therapeutic agents, the dosage of the traditional Chinese medicine composition is either less than or not less than the effective dose; the dosage of the other therapeutic agents is either less than or not less than the effective dose.

[0032] In this invention, the effective dose includes a "therapeutic effective dose" or a "preventive effective dose," which refers to an amount sufficient, within reasonable medical judgment, to treat or prevent a patient's disease while avoiding serious side effects with a sufficiently low risk-benefit ratio. The preventive or therapeutic effective dose of a drug will vary depending on factors such as the specific drug chosen (e.g., considering the drug's potency, effectiveness, and half-life), the chosen route of administration, the disease being prevented or treated, the severity of the disease, the patient's age, size, weight, and physical condition, the patient's medical history, the duration of prevention or treatment, the nature of concurrent therapies, and the desired preventive or therapeutic effect, but can still be routinely determined by those skilled in the art.

[0033] In this invention, prevention and / or treatment refer to measures targeting the inflammatory response to viral infection. This includes methods for preventing the occurrence of an inflammatory response to viral infection, the purpose of which is to prevent, mitigate (reduce) undesirable physiological changes or lesions in the treated subject. Beneficial or desired clinical outcomes include, but are not limited to, avoiding the occurrence of an inflammatory response to viral infection, reducing the symptoms of an inflammatory response to viral infection, weakening the degree of an inflammatory response to viral infection, stabilizing the inflammatory state of an inflammatory response to viral infection (i.e., not worsening), delaying or slowing the progression of an inflammatory response to viral infection, improving or alleviating the inflammatory state of an inflammatory response to viral infection, and remission (whether partial or complete remission), whether detectable or undetectable. Subjects requiring treatment include those who already have the condition or disease, those who are susceptible to the condition or disease, or those who intend to prevent the condition or disease.

[0034] In this invention, the target of prevention and control is humans or mammals, including bovines, equines, sheep, pigs, canines, felines, rodents, and primates.

[0035] Compared with the prior art, the present invention has at least the following beneficial effects: The traditional Chinese medicine composition of this invention includes areca nut, magnolia bark, cardamom, anemarrhena rhizome, white peony root, scutellaria root, licorice root, honeysuckle flower, forsythia fruit, malt, hawthorn fruit, medicated leaven, and tangerine peel. This invention found that administration of this traditional Chinese medicine composition significantly increased the IL-10 content in the bronchoalveolar lavage fluid of mice, while significantly decreasing the levels of IL-1β, TNF-α, and IL-6. This indicates that the traditional Chinese medicine composition can reduce lung tissue inflammatory damage by inhibiting the release of pro-inflammatory cytokines such as IL-1β, TNF-α, and IL-6, and promoting the content of the anti-inflammatory cytokine IL-10. This demonstrates that the traditional Chinese medicine composition has a good therapeutic effect in antiviral inflammatory responses. Moreover, this traditional Chinese medicine composition is an existing prepared Chinese medicine, safe to use, and its preparation method is simple, rapid, efficient, and easy to operate. The prepared decoctions, granules, capsules, tablets, mixtures, pills, tinctures, syrups, or sprays are easy to carry and convenient to take. Attached Figure Description

[0036] Figure 1 The image shows the (200×) results of the effects of LPS on the pathological characteristics of mouse lung tissue in Example 2 of this invention. Detailed Implementation

[0037] This invention provides traditional Chinese medicine compositions, preparations, preparation methods, and their application in the preparation of drugs for antiviral inflammatory responses. Those skilled in the art can refer to the content of this document and appropriately modify the process parameters to achieve the desired results. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in this invention. The methods and applications of this invention have been described through preferred embodiments. Those skilled in the art can clearly modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit, and scope of this invention to realize and apply the technology of this invention.

[0038] The test materials used in this invention are all common commercial products and can be purchased on the market.

[0039] This invention proposes the application of a traditional Chinese medicine composition in the preparation of antiviral inflammatory response drugs.

[0040] The traditional Chinese medicine composition includes: 1-20 parts of areca nut, 1-20 parts of magnolia bark, 1-20 parts of cardamom, 1-20 parts of anemarrhena rhizome, 1-20 parts of white peony root, 1-20 parts of scutellaria root, 1-20 parts of licorice root, 1-20 parts of honeysuckle flower, 1-20 parts of forsythia fruit, 1-20 parts of malt, 1-20 parts of hawthorn fruit, 1-20 parts of medicated leaven, and 1-20 parts of dried tangerine peel.

[0041] The above-mentioned uses can be any method beneficial to improving the corresponding symptoms of the patient, including treatment or prevention. The compositions of the present invention can be directly ground into powder, or can be extracts obtained by conventional means in the art. The traditional Chinese medicines used in the compositions of the present invention can also be used by directly grinding into powder, extracts, or other processed forms.

[0042] Most preferably, the herbal composition includes: 9 parts areca nut, 9 parts magnolia bark, 6 parts cardamom, 9 parts anemarrhena rhizome, 9 parts white peony root, 9 parts scutellaria root, 6 parts licorice root, 9 parts honeysuckle flower, 9 parts forsythia fruit, 9 parts malt, 9 parts hawthorn fruit, 9 parts medicated leaven, and 6 parts dried tangerine peel. The processing specifications for malt, hawthorn fruit, and medicated leaven are charred malt, charred hawthorn fruit, and charred medicated leaven.

[0043] Specifically, antiviral inflammatory response drugs are selected from oral, injectable, or topical formulations.

[0044] Specifically, antiviral inflammatory response drugs are selected from decoctions, granules, capsules, tablets, mixtures, pills, tinctures, syrups, or sprays.

[0045] The above-mentioned composition of the present invention can be made by directly grinding the ingredients into powder using traditional methods, or by obtaining extracts through conventional means, so as to prepare different dosage forms, achieve better therapeutic effects, and be more conducive to the preparation of modern pharmaceutical dosage forms.

[0046] This invention also proposes a method for preparing any of the aforementioned traditional Chinese medicine compositions, the preparation method being as follows: Areca nut, magnolia bark, cardamom, anemarrhena rhizome, white peony root, scutellaria root, licorice root, honeysuckle flower, forsythia fruit, malt, hawthorn fruit, medicated leaven, and dried tangerine peel are decocted twice with water, 11 times the amount of water for the first decoction and 9 times the amount of water for the second decoction, each decoction lasting 1.5 hours. The herbs are soaked for 30 minutes before the first decoction. The decoctions are combined, filtered, and the filtrate is concentrated under reduced pressure to a relative density of approximately 1.10 (50℃~60℃). The filtrate is then spray-dried into a dry extract powder. Maltodextrin is added, mixed well, and granulated to obtain the final product.

[0047] The compound drug for antiviral infection and inflammatory response of this invention can significantly increase the IL-10 content in bronchoalveolar lavage fluid of mice and significantly decrease the content of IL-1β, TNF-α, and IL-6. This indicates that the traditional Chinese medicine composition can reduce lung tissue inflammatory response damage by inhibiting the release of pro-inflammatory cytokines IL-1β, TNF-α, and IL-6 and promoting the content of the anti-inflammatory cytokine IL-10. This demonstrates that the traditional Chinese medicine composition has a good therapeutic effect in the inflammatory response of antiviral infection. Moreover, this traditional Chinese medicine composition is an existing prepared Chinese medicine, safe to use, and its preparation method is simple, rapid, efficient, and easy to operate. The prepared mixture, tablets, capsules, micro-pellets, etc., are easy to carry and convenient to take.

[0048] Unless otherwise specified, all experiments in the following experiments were conducted under standard conditions or conditions recommended by the manufacturer. Active pharmaceutical ingredients (APIs) or excipients, as well as reagents or instruments whose manufacturers are not specified, are all commercially available products. Unless otherwise stated, all percentages, ratios, proportions, or parts are by weight.

[0049] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to this invention. The invention is further illustrated below with reference to embodiments.

[0050] <Example 1>: Preparation of Traditional Chinese Medicine Compositions and Their Preparations Areca nut (9 parts), Magnolia officinalis (9 parts), Amomum tsao-ko (6 parts), Anemarrhena asphodeloides (9 parts), Paeonia lactiflora (9 parts), Scutellaria baicalensis (9 parts), Glycyrrhiza uralensis (6 parts), Lonicera japonica (9 parts), Forsythia suspensa (9 parts), roasted malt (9 parts), roasted hawthorn (9 parts), roasted Shenqu (9 parts), and dried tangerine peel (6 parts). These nine ingredients are decocted twice with water. The first decoction uses 11 times the amount of water, and the second uses 9 times the amount of water. Each decoction lasts 1.5 hours. The ingredients are soaked for 30 minutes before the first decoction. The decoctions are combined, filtered, and the filtrate is concentrated under reduced pressure to a relative density of approximately 1.10 (50℃~60℃). The filtrate is then spray-dried into a dry extract powder to obtain the intermediate of the composition. An appropriate amount of maltodextrin is added, mixed well, and granulated to obtain the final product.

[0051] <Comparative Example 1> 6 parts of Areca catechu L., 3 parts of Magnolia officinalis Rehd. et Wils., 1.5 parts of Tsaoko fruit, 3 parts of Anemarrhena asphodeloides Bunge, 3 parts of Paeonia lactiflora Pall., 3 parts of Scutellaria baicalensis Georgi, and 1.5 parts of Glycyrrhiza uralensis Fisch. There are seven herbs in total. Decoct twice with water. Add 11 times the amount of water for the first time and 9 times the amount of water for the second time. Decoct for 1.5 hours each time. Soak for 30 minutes before the first decoction. Combine the decoction, filter, and concentrate the filtrate under reduced pressure to a relative density of about 1.10 (50 °C - 60 °C), and spray dry to obtain dry extract powder, getting the composition intermediate. Add an appropriate amount of maltodextrin, mix evenly, and make granules to obtain the product.

[0052] Experimental Example 2: Pharmacological Test Pharmacodynamic Study of the Test Substance on LPS-Induced Acute Lung Injury in Mice 1 Experimental Materials 1.1 Animals SPF-grade C57BL / 6J mice, weighing 18 - 25 g, 60 in total, with half males and half females, sourced from Jinan Pengyue Experimental Animal Breeding Co., Ltd. Housed in the animal facilities of Stander Kechuang Pharmaceutical Technology (Qingdao) Co., Ltd. (Drug Safety Evaluation Center) (Experimental Animal Use License No.: SYXK (Lu) 2024 0016), controlled room temperature: 20 - 26 °C, controlled humidity: 30% - 70%, lighting time: 12 hours of light and 12 hours of darkness, feeding method: cage feeding, 2 - 5 mice per cage.

[0053] 1.2 Drugs Table 1. Table of Test Substances

[0054] 1.3 Instruments Table 2. Table of Instrument Information

[0055] 2 Dosage Design In this experiment, the high dose was set at 30 g crude drug / kg (about 2 times the clinical equivalent dose); with a group distance of 2 times, the medium dose was set at 15 g crude drug / kg (equivalent to the human clinical equivalent dose), and the low dose was set at 7.5 g crude drug / kg (equivalent to 1 / 2 times the human clinical equivalent dose).

[0056] High-dose solution of the test substance (3 g crude drug / mL): Weigh the dry extract powder of the test substance and place it in a centrifuge tube, add a certain volume of 0.9% sodium chloride injection solution and stir well to obtain.

[0057] Medium-dose solution of the test substance (1.5 g crude drug / mL): Take a certain volume of the high-dose solution and place it in a centrifuge tube, add an equal volume of 0.9% sodium chloride injection solution and shake well to obtain.

[0058] Low-dose solution of the test substance (0.75 g crude drug / mL): Take a certain volume of medium-dose solution and place it in a centrifuge tube. Add an equal volume of 0.9% sodium chloride injection and shake thoroughly to mix.

[0059] High-dose comparative solution (3g crude drug / mL): Weigh an appropriate amount of the comparative substance dry extract powder and place it in a centrifuge tube. Add a certain volume of 0.9% sodium chloride injection and stir thoroughly to obtain the solution.

[0060] 3 Experimental Methods 3.1 Trial grouping and dosing design Eighty-four qualified C57BL / 6 mice were selected for the experiment and randomly divided into four groups (n=14 each, half male and half female) according to weight and sex: a normal control group, a model control group, low-, medium-, and high-dose test substance groups, and a control group. The low-, medium-, and high-dose test substance groups and the control group were administered different concentrations of the dry extract powder by gavage, while the normal control group and the model control group were administered 0.9% sodium chloride injection by gavage. The dosage was 10 mL / kg. Administered once daily for three consecutive days. Details of the experimental groups and administration are shown in Table 3 below.

[0061] Table 3. Trial grouping and dosing design

[0062] 3.2 Model Establishment Thirty minutes after the last administration, mice in both the model control group and the treatment group were anesthetized with an intraperitoneal injection of 1% sodium pentobarbital (50 mg / kg). The mice were placed abdomen-up, with their heads fixed to a self-made cardboard. After shaving the neck hair, the surgical site was disinfected with iodine, and a longitudinal incision was made in the neck to expose the trachea. Approximately 50 μL of the prepared LPS solution (4 mg / ml) was drawn into a 0.3 mL syringe, and the needle was inserted at approximately a 45° angle to the tracheal wall. The entire LPS solution was slowly dripped into the trachea, and the mouse was immediately held upright for approximately 1 minute before the wound was sutured. The normal control group received an equal volume of 0.9% sodium chloride injection intratracheally.

[0063] 4 indicators to observe 4.1 Detection of cytokine content in bronchoalveolar lavage fluid Twenty-four hours after modeling, the first eight mice in each group (half male and half female) were euthanized by cervical dislocation. The thoracic cavity was opened, the trachea was exposed and separated, and an oblique incision was made. A self-made flat-tipped needle was inserted into the trachea and fixed with surgical sutures (to prevent air leakage). Approximately 0.5 mL of 0.9% sodium chloride injection solution was drawn into a 1 mL syringe and used to lavage the lungs. This was repeated three times each time, and the lavage was repeated three times. The bronchoalveolar lavage fluid (BALF) obtained was combined and centrifuged at 1500 rpm and 4°C for 10 min. The supernatant was collected and stored at -20°C or -80°C for later use. The levels of IL-1β, TNF-α, IL-6, and IL-10 in the BALF were determined by ELISA.

[0064] 4.2 Pathological examination 4.2.1 Lung wet weight and lung wet / dry weight ratio Twenty-four hours after modeling, six mice in each group were euthanized by cervical dislocation. The thoracic cavity was opened, and lung tissue was harvested. The condition of the lung lesions was observed visually, and the wet weight was measured. The left lung was ligated and removed. After blotting the tissue fluid and blood on the surface of the removed left lung with filter paper, the wet weight of the left lung was measured, and it was placed in a 60°C constant temperature incubator to dry for 72 hours. After drying, the lung was removed and the dry weight was measured to determine the wet / dry weight ratio of the lung and assess the degree of pulmonary edema.

[0065] 4.2.2 Histopathological examination The remaining right lung was perfused and preserved with neutral formalin solution, then routinely dehydrated, embedded, sectioned, and stained with hematoxylin and eosin (HE). The pathological changes, such as inflammatory cell infiltration and tissue lesions, were observed under a microscope.

[0066] 5. Data Statistical Analysis Experimental data were analyzed using SPSS and expressed as mean ± standard deviation. The data were first tested for homogeneity of variance using Levene's test. If the variances were homogeneous (p>0.05), one-way ANOVA was performed. If the variances were unequal (p≤0.05), Dunnett's T3 test was used to compare between groups.

[0067] 6. Test Results 6.1 Effects on cytokine levels in mouse bronchoalveolar lavage fluid Compared with the normal control group, the levels of IL-1β, TNF-α, and IL-6 in the bronchoalveolar lavage fluid of mice in the model control group were significantly increased (P<0.05). Compared with the model control group, the levels of IL-1, TNF-α, and IL-6 in the bronchoalveolar lavage fluid of mice in the low-, medium-, and high-dose groups of the test substance and the control group were all significantly decreased (P<0.05). See Table 4 below for details.

[0068] Table 4. Results of Cytokine Level Detection

[0069] Note: Compared with the blank control group, ** indicates P<0.01; compared with the model control group, & indicates P<0.05, && indicates P<0.01.

[0070] Compared with the blank control group, the levels of IL-1β, TNF-α, and IL-6 in the bronchoalveolar lavage fluid of mice in the model control group were significantly increased (P<0.01), while the level of IL-10 was significantly decreased (P<0.01).

[0071] Compared with the model control group, the IL-10 content in the bronchoalveolar lavage fluid of mice in the low, medium and high dose groups of the test substance was significantly increased (P<0.01) and showed a certain dose dependence; the IL-1β, TNF-α and IL-6 content in the bronchoalveolar lavage fluid of mice in the medium and high dose groups of the test substance was significantly decreased (P<0.05) and showed a certain dose dependence.

[0072] In the control group of mice, the levels of IL-1β, TNF-α, and IL-6 in bronchoalveolar lavage fluid were significantly decreased (P<0.05), while the level of IL-10 was significantly increased (P<0.05). The control group, as a high-dose administration, showed effects comparable to the low-dose test substance group.

[0073] As can be seen from the above, intratracheal instillation of LPS can induce a large release of pro-inflammatory cytokines in lung tissue. However, administration of both the test substance and the control substance can inhibit the release of pro-inflammatory cytokines in lung tissue and promote the release of anti-inflammatory cytokines. Furthermore, the inhibitory and promoting effects of the test substance on pro-inflammatory and anti-inflammatory cytokines in bronchoalveolar lavage fluid are dose-dependent.

[0074] The test substance was more effective than the control substance.

[0075] 6.2 Pathological examination 6.2.1 Lung wet weight and lung wet / dry weight ratio Lung wet weight: Compared with the normal control group, the lung wet weight of the model control group was significantly increased (p<0.05); compared with the model control group, the lung wet weight of the medium-dose group, the high-dose group and the control group of the test substance were all significantly decreased (p<0.05).

[0076] Lung wet / dry weight ratio: Compared with the normal control group, the lung wet / dry weight ratio of the model control group was significantly increased (p<0.05); compared with the model control group, the lung wet weight of the low-dose group, medium-dose group, high-dose group and control group of the test substance was significantly decreased (p<0.05).

[0077] In summary, except for the low dose of the test substance which failed to effectively reduce lung wet weight, the other dose groups of the test substance effectively reduced lung wet weight and lung wet / dry weight ratio, suggesting that the dry extract powder of the test substance can reduce lung tissue edema.

[0078] The control group of the comparative substance showed a similar reduction in lung wet weight and lung wet / dry weight ratio as the medium-dose group of the test substance.

[0079] Table 5. Results of lung wet weight and lung wet / dry weight ratio

[0080] 6.2.2 Histopathological examination The lungs of each group were observed under a microscope; photographs are shown below. Figure 1 . Figure 1 In the diagram, a~f represent the normal control group, the model control group, the control group of the reference substance, the low-dose group of the test substance, the medium-dose group of the test substance, and the high-dose group of the test substance, respectively.

[0081] from Figure 1 It can be seen that, under a microscope, the lungs of the normal control group animals showed clear and intact lobular structures, full alveoli, and no abnormalities in the pleura covering the lung surface. No abnormalities were found in the structures of bronchi, alveolar ducts, alveoli, etc., and the interstitium around the bronchi and blood vessels in the lung parenchyma. Compared with the normal control group, under a microscope, the lungs of the model control group animals showed alveolar and interstitial inflammatory cell infiltration, accompanied by hemorrhage and edema, with 1 / 6 of them being mild and 5 / 6 being moderate.

[0082] Compared with the model control group, microscopic observation of the lungs of animals in the low-dose, medium-dose, and high-dose groups of the test substance and the control group also revealed alveolar and interstitial inflammatory cell infiltration, accompanied by hemorrhage and edema. In the low-dose group of the test substance, 2 / 6 animals had mild symptoms, 1 / 6 animals had mild symptoms, and 3 / 6 animals had moderate symptoms. In the medium-dose group of the test substance, 3 / 6 animals had mild symptoms, 2 / 6 animals had mild symptoms, and 1 / 6 animals had moderate symptoms. In the high-dose group of the test substance, 3 / 6 animals had mild symptoms and 3 / 6 animals had mild symptoms. In the control group of the control substance, 2 / 6 animals had mild symptoms, 1 / 6 animals had mild symptoms, and 2 / 6 animals had moderate symptoms.

[0083] In summary, except for the normal control group, all other groups of animals showed alveolar / interstitial inflammatory cell infiltration in their lungs, with or without hemorrhage / edema. Compared with the model control group, the severity of lung lesions was reduced to varying degrees in the low / medium / high dose groups of the test substance and the control group of the comparative substance, with the high dose group of the test substance showing the greatest reduction; the reduction in the control group of the comparative substance was less than that in the high dose group of the test substance.

[0084] Table 6. Results of lesion severity

[0085] 7 Conclusions The experimental results showed that 24 hours after intratracheal instillation of LPS, the levels of IL-1β, TNF-α, and IL-6 in the bronchoalveolar lavage fluid of the model control group animals were significantly increased, as were the lung wet weight and lung wet / dry weight ratio. Mild to moderate inflammatory cell infiltration was observed in the alveoli and interstitium, accompanied by hemorrhage and edema. This indicates that intratracheal instillation of 5 mg / kg LPS induced significant pulmonary edema and a severe inflammatory response, successfully replicating the ALI animal model.

[0086] After intervention with the test substance dry extract powder, low, medium, and high doses of the test substance significantly increased the IL-10 content in the bronchoalveolar lavage fluid of mice; medium and high doses of the test substance significantly decreased the IL-1β, TNF-α, and IL-6 content in the bronchoalveolar lavage fluid of mice. This suggests that the test substance can reduce lung tissue inflammatory damage by inhibiting the release of pro-inflammatory cytokines IL-1β, TNF-α, and IL-6 and promoting the level of the anti-inflammatory cytokine IL-10. Compared with the test substance, the control substance showed a reduced effect in inhibiting the inflammatory response.

[0087] Except for the low dose of the test substance, which failed to effectively reduce lung wet weight, all other dose groups of the test substance effectively reduced lung wet weight and the lung wet / dry weight ratio, suggesting that the dry extract of the test substance can reduce pulmonary edema. The severity of lung lesions was reduced to varying degrees in the low / medium / high dose groups of the test substance, with the high dose group showing the greatest reduction. The control substance showed a reduced effect compared to the test substance.

[0088] In summary, intratracheal instillation of LPS induces a large release of pro-inflammatory cytokines from lung tissue, while administration of the test substance inhibits the release of pro-inflammatory cytokines and promotes the release of anti-inflammatory cytokines. Furthermore, the inhibitory and promoting effects of the test substance on pro-inflammatory and anti-inflammatory cytokines in bronchoalveolar lavage fluid are dose-dependent. The comparative substance showed a reduced effect compared to the test substance.

[0089] The above are merely preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A traditional Chinese medicine composition, characterized in that, It is made from the following raw materials in parts by weight: 1-20 parts areca nut, 1-20 parts magnolia bark, 1-20 parts cardamom, 1-20 parts anemarrhena rhizome, 1-20 parts white peony root, 1-20 parts scutellaria root, 1-20 parts licorice root, 1-20 parts honeysuckle, 1-20 parts forsythia fruit, 1-20 parts malt, 1-20 parts hawthorn, 1-20 parts medicated leaven, and 1-20 parts dried tangerine peel.

2. The traditional Chinese medicine composition according to claim 1, characterized in that: in, The traditional Chinese medicine composition is made from the following raw materials in parts by weight: 9 parts areca nut, 9 parts magnolia bark, 6 parts cardamom, 9 parts anemarrhena rhizome, 9 parts white peony root, 9 parts scutellaria root, 6 parts licorice root, 9 parts honeysuckle flower, 9 parts forsythia fruit, 9 parts malt, 9 parts hawthorn fruit, 9 parts medicated leaven, and 6 parts dried tangerine peel.

3. The traditional Chinese medicine composition according to claim 1, characterized in that: in, In the aforementioned traditional Chinese medicine composition, the processing specifications for malt, hawthorn, and Shenqu (a type of medicinal fermentation) are charred malt, charred hawthorn, and charred Shenqu.

4. A traditional Chinese medicine preparation, characterized in that, It is mainly made from the traditional Chinese medicine composition according to any one of claims 1 to 3.

5. The traditional Chinese medicine preparation according to claim 4, characterized in that: in, The traditional Chinese medicine preparation is made from the traditional Chinese medicine composition and pharmaceutically acceptable excipients.

6. The traditional Chinese medicine preparation according to claim 4, characterized in that: in, The dosage form of the traditional Chinese medicine preparation is any one of the following: decoction, granules, capsules, tablets, compound preparations, pills, tinctures, syrups, or sprays.

7. A method for preparing a traditional Chinese medicine preparation as described in any one of claims 4 to 6, characterized in that: Areca nut, magnolia bark, cardamom, anemarrhena rhizome, white peony root, scutellaria root, licorice root, honeysuckle flower, forsythia fruit, malt, hawthorn fruit, medicated leaven, and dried tangerine peel are decocted in water. The decoction is then filtered, concentrated, and dried to obtain the final product. During decoction, the mass ratio of water to the herbal medicine composition is (4-40):1; Boil 1 to 3 times, 1 to 2 hours each time.

8. The method for preparing the traditional Chinese medicine preparation according to claim 7, characterized in that: in, Areca nut, magnolia bark, cardamom, anemarrhena rhizome, white peony root, scutellaria root, licorice root, honeysuckle flower, forsythia fruit, malt, hawthorn fruit, medicated leaven, and tangerine peel were decocted twice with water, 11 times the amount of water for the first decoction and 9 times the amount of water for the second decoction. Each decoction lasted 1.5 hours. The ingredients were soaked for 30 minutes before the first decoction. The decoctions were combined, filtered, and the filtrate was concentrated under reduced pressure to an extract with a relative density of 1.05 to 1.30 at 50℃ to 60℃. The extract was then concentrated and dried.

9. The method for preparing a traditional Chinese medicine preparation according to claim 8, characterized in that: in, The drying process employs spray drying, and subsequently includes the addition of maltodextrin, mixing, and granulation.

10. The use of the traditional Chinese medicine composition according to any one of claims 1 to 3 or the traditional Chinese medicine preparation according to any one of claims 4 to 6 in the preparation of a medicament for antiviral infection and inflammatory response.

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