Application of Keke preparation in preparation of medicine for treating cough and airway and lung tissue injury after influenza virus infection
By developing a traditional Chinese medicine composition Kemp preparation based on Maxing Shigan Decoction in "Treatise on Febrile Diseases", the treatment problems of coughing after influenza virus infection and airway and lung tissue damage have been solved, and good antiviral and symptom-improving effects have been achieved, and high safety and stability have been achieved.
Patent Information
- Application Number
- CN202411863011.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-17
- Publication Date
- 2025-05-16
AI Technical Summary
The prior art is difficult to effectively treat cough and airway and lung tissue damage after influenza virus infection, and common Western medicines have adverse reactions and drug resistance problems.
A traditional Chinese medicine composition, a preparation for cough-cough, was developed based on Maxing Shigan Decoction in "Treatise on Febrile Diseases", and added gypsum, raspberry seeds and poppy shells to form ephedra and bitter almonds as the monarch, platycodon and raspberry seeds as the minister, and licorice and poppy shells as the adjuvant drugs, which are used to treat cough after influenza virus infection and damage to the airway and lung tissue.
Keke preparations show good antiviral effects, which can improve cough symptoms after infection and damage to airway and lung tissues. Compared with the positive drug Asme group, Keke tablets have more stable antiviral, inhibit cough and improve inflammation in the body, and are basically non-toxic.
Smart Images
Figure CN120000718A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the field of new drug applications, and in particular to the application of a cough suppressant preparation in the preparation of drugs for treating cough and airway and lung tissue damage after influenza virus infection. Background Art
[0002] At present, the clinical manifestation frequency, misdiagnosis rate and morbidity of postinfectious cough (PIC) are high, which has brought a heavy economic burden to the society and led to a large waste of medical resources. Reports point out that when the cough frequency is too high and the duration is too long, patients may experience complications such as vomiting, rib fractures, urinary incontinence, syncope, muscle pain, fatigue and depression. Therefore, the development of effective PIC treatment drugs is very necessary to solve people's health problems and alleviate economic pressure.
[0003] At present, there are different methods for treating PIC in clinical practice. Commonly used Western medicine drugs include cephalosporins, macrolides, leukotriene receptor antagonists, budesonide, ipratropium bromide and ambroxol hydrochloride. Although these Western medicines can relieve symptoms, they impose a heavy economic burden on patients, and these drugs are often accompanied by adverse reactions, such as drowsiness, dizziness, loss of appetite, and viral resistance. The main reason for these shortcomings and limitations is the high cost of Western medicine research and development. Although these drugs can relieve the symptoms of post-infection cough caused by influenza virus, they cannot provide specific antiviral treatment for cough caused by influenza virus. Summary of the invention
[0004] In view of the deficiencies of the prior art, the purpose of the present invention is to provide a cough suppressant preparation for use in the preparation of a drug for treating cough and airway and lung tissue damage after influenza virus infection. The cough suppressant preparation has a good antiviral effect and can improve cough and airway and lung tissue damage after influenza virus infection.
[0005] In order to achieve this purpose, the present invention targets the TCM characterization of cough after infection caused by influenza virus, and develops a new application of a Chinese medicine composition, Keke preparation, for treating cough after influenza virus infection. The present invention also constructs an in vivo model of cough after infection in guinea pigs and combines it with in vitro antiviral experiments to study the efficacy and material basis of Keke preparation in cough after influenza virus infection. This study not only comprehensively evaluates the efficacy of Keke preparation in treating cough after influenza virus infection, but also further studies its material basis and potential pharmacological mechanism.
[0006] The Keke preparation of the present invention is a traditional Chinese medicine composition, which is developed on the basis of the famous prescription of Ma Xing Shi Gan Tang for treating lung heat cough and asthma in "Treatise on Febrile Diseases" by the medical saint Zhang Zhongjing. The effect of Ma Xing Shi Gan Tang is pungent and cool, clearing the lungs and relieving asthma, and is used for exogenous wind evil and pathogenic heat blocking the lungs. The components of the Keke tablets include ephedra, platycodon, bitter almond, liquorice, gypsum, radish seeds and poppy shells, and gypsum, radish seeds and poppy shells are added to the original prescription. In the cough-killing tablets, ephedra and bitter almond are the main drugs. Ephedra can clear the lung qi and dispel cold, while bitter almond has the function of purging and descending qi, relieving cough and asthma. The two are used together to dispel wind and cold, clear the lung and relieve cough; Platycodon grandiflorum leads the medicine upward, and has the function of clearing the lung, removing phlegm and relieving cough, while radish seeds descend qi and resolve phlegm, and they are both ministerial drugs, assisting the main drug to strengthen the function of clearing the lung, removing phlegm and relieving cough; poppy shell has the function of astringing the lung and relieving cough, and ephedra is used to prevent excessive purgation, so it is an adjuvant. Poppy shell is combined with ephedra, one is astringent and the other is astringent, they can clear the obstruction of evil qi and restrain the dissipation of the lung; gypsum is bitter and cold, and a large amount of it can clear heat and purge fire, which can easily damage the spleen and stomach. A small amount used as an adjuvant can clear and relieve stagnant heat to prevent wind and cold, and phlegm from turning into heat for a long time; licorice can both moisten the lung and relieve cough, and can harmonize various drugs, so it is a guiding drug. The whole prescription is pungent, sweet, bitter and moist, with appropriate dispersing and suppressing, and moderate dispersing and astringing, which can dispel wind and cold, regulate qi and resolve phlegm, and restrain the lungs and relieve cough. Clinical data show that Keke tablets have the effects of dispersing lungs and clearing heat, relieving cough and relieving asthma. According to the "Pharmacopoeia of the People's Republic of China: 2020 Edition", the prescription of Keke preparation is: 360g of ephedra, 360g of poppy shell, 360g of licorice, 360g of bitter almond, 112.5g of radish seeds, 112.5g of platycodon, and 112.5g of gypsum. At present, the above-mentioned Keke preparations are mostly used as medicines for wind-heat, wind-cold, and wind-cold with phlegm and dampness cough, but there are no reports on the effect of Keke preparations in treating cough after influenza virus infection and airway and lung tissue damage.
[0007] Specifically, to achieve the purpose of the present invention, the present invention provides the use of a cough suppressant preparation in the preparation of a medicine for treating cough after influenza virus infection, wherein the cough suppressant preparation comprises the following components in parts by weight: 360 parts of ephedra, 360 parts of poppy shells, 360 parts of liquorice, 360 parts of bitter almonds, 112.5 parts of radish seeds, 112.5 parts of platycodon and 112.5 parts of gypsum.
[0008] To achieve the purpose of the present invention, the present invention also provides the use of a cough suppressant in the preparation of a drug for treating airway or lung tissue damage after influenza virus infection, wherein airway and lung tissue damage after influenza virus infection may be accompanied by cough symptoms or without cough symptoms, and the cough suppressant comprises the following components in parts by weight: 360 parts of ephedra, 360 parts of poppy shells, 360 parts of liquorice, 360 parts of bitter almonds, 112.5 parts of radish seeds, 112.5 parts of platycodon and 112.5 parts of gypsum.
[0009] In some embodiments of the present invention, the cough suppressant preparation is an oral dosage form.
[0010] In some embodiments of the present invention, the oral dosage form is a tablet, granule, powder, paste, capsule, pill or oral liquid preparation.
[0011] In some embodiments of the present invention, the cough suppressant preparation is a tablet, and the preparation method of the cough suppressant preparation is to take 360 parts of ephedra, 360 parts of poppy shell, 360 parts of licorice, 360 parts of bitter almond, 112.5 parts of radish seeds, 112.5 parts of platycodon and 112.5 parts of gypsum, crush the ephedra and poppy shell powder into fine powder respectively, sieve, and keep 165 parts of fine powder for standby use; adjust the pH value of the remaining ephedra and poppy shell coarse powder to 4.5-5.5 with an acidic solution, and fry Boil twice, each time for 2 hours, filter, adjust the pH value of the filtrate to 6.5-7.5 with alkaline solution, and keep the medicinal solution for later use; add liquorice, bitter almond, radish seed, platycodon and gypsum into water and boil twice, each time for 1 hour, filter, combine the filtrate with the above solution, and concentrate under reduced pressure to a clear paste with a relative density of 1.20-1.26, add fine powder of ephedra and poppy shell, mix well, dry, crush into fine powder, add auxiliary materials, mix well, granulate, dry, press into tablets, and coat with film.
[0012] Compared with the prior art, the present invention can achieve the following beneficial effects: the Keke preparation of the present invention is used to treat cough after infection, and has good in vitro antiviral activity, in vivo improvement of guinea pig cough symptoms and anti-inflammatory and antiviral efficacy. Combining in vitro cell experiments with in vivo guinea pig experiments, it was found that compared with the positive drug Asmi group, the Keke tablet intervention group had more stable in vivo antiviral, post-infection cough symptom inhibition, and tissue inflammation and damage efficacy, and was basically non-toxic. The new use of the Keke preparation of the present invention for treating cough after influenza virus infection has good social and economic benefits. BRIEF DESCRIPTION OF THE DRAWINGS
[0013] Figure 1 This is a statistical chart showing the effect of Keke tablets on MDCK cell viability.
[0014] Figure 2 Statistical chart showing that Keke tablets improve the cell viability of MDCK cells after influenza virus infection.
[0015] Figure 3 This is a plaque image showing that Keke tablets inhibit influenza virus replication in MDCK cells.
[0016] Figure 4 This is the cough waveform of guinea pigs treated with Keke tablets for cough following influenza virus infection.
[0017] Figure 5 This is a statistical chart of the efficacy of Keke tablets in treating cough in guinea pigs infected with influenza virus.
[0018] Figure 6 Mass spectrometry analysis of the serum material basis of Keke tablets in the treatment of post-infectious cough in guinea pigs.
[0019] Figure 7 This is a network pharmacology analysis of the serum material basis of Keke tablets.
[0020] Figure 8 The following are pathological images showing that Keke Tablets can improve bronchial (A) and lung tissue damage (B) in guinea pigs.
[0021] Fig. 9 The changes in body weight (A), lung index (B), spleen index (C) and bronchial index (D) in patients with post-infectious cough treated with Keke Tablets.
[0022] Fig.10 Heat map analysis of the effect of Shike Tablets on improving blood routine in bronchoalveolar lavage fluid of guinea pigs.
[0023] Fig.11 This is a statistical chart showing the effect of Keke tablets in inhibiting the secretion of pro-inflammatory factors in guinea pig lung tissue.
[0024] Fig.12 Micrograph (A) and viral load statistics (B) of the inhibition of viral load in guinea pig lung tissue after influenza virus infection by Shike Tablets.
[0025] Fig.13 This is the immunofluorescence image of influenza virus NP protein in guinea pig lung tissue after Keke tablets inhibited influenza virus infection.
[0026] The present invention is further described in detail below with reference to the accompanying drawings and specific implementation modes. DETAILED DESCRIPTION
[0027] The prescription of the cough-killing preparation used in the embodiment of the present invention is 360g of ephedra, 360g of poppy shell, 360g of liquorice, 360g of bitter almond, 112.5g of radish seed, 112.5g of platycodon and 112.5g of gypsum. The sources of the above Chinese medicinal materials are consistent with the Chinese Pharmacopoeia. The preparation method is as follows: take the above seven flavors, crush ephedra and poppy shell into fine powder, sieve, keep 165g of each fine powder for use; the remaining coarse powder is decocted twice with an acidic aqueous solution (adjust the pH value to about 5 with a 10% hydrochloric acid solution), each time for 2 hours, filtered, and the filtrate is adjusted to a pH value of 7 with a 10% sodium hydroxide solution, and the liquid is used for use; the remaining five flavors such as licorice are decocted twice with water, each time for 1 hour, filtered, the filtrate is combined with the above solution, and concentrated under reduced pressure to a clear paste with a relative density of 1.20-1.26 (60°C), add ephedra and poppy shell fine powder, mix, dry, crush into fine powder, add appropriate amount of auxiliary materials, mix, granulate, dry, press into 1000 tablets (specification 1) or 1500 tablets (specification 2), and film-coat. The following test is carried out with the specification 1 gram cough tablet.
[0028] Test example
[0029] Experiment 1 In vitro test of cell viability and antiviral efficacy of Keke tablets
[0030] To evaluate the in vitro effects of Keke tablets on cell viability and antiviral efficacy, the drug was ground and dissolved in DMSO solution to prepare Keke tablet solutions of different concentrations (200ug / ml, 150ug / ml, 75ug / ml, 50ug / ml, 37.5ug / ml, 25ug / ml, 10ug / ml, 5ug / ml, 2.5ug / ml, 1.25ug / ml, 0.625ug / ml). MDCK cells in the logarithmic growth phase were cultured at a rate of 1x10 4 The cells were inoculated at a density of 100 μg / mL in a 96-well cell culture plate. When the cell confluence reached 75%, the cells were given Keke tablets and incubated for 24 hours. The cell activity was detected by MTT (tetrazolium salt colorimetry). Figure 1 As shown, the results showed that Keke tablets had basically no cytotoxicity.
[0031] On this basis, MDCK cells were infected with 1MOI A / PR / 8 virus and given different concentrations of Keke tablets (5ug / ml, 10ug / ml, 12.5ug / ml, 20ug / ml, 25ug / ml, 40ug / ml, 50ug / ml, 75ug / ml). After 48 hours, the cell activity was detected by MTT. Figure 2 As shown, the results showed that Keke tablets had good in vitro antiviral effects, especially at 40ug / ml.
[0032] Test of the antiviral efficacy of 2g cough tablets
[0033] MDCK cells were infected with 1MOI A / PR / 8 virus and given different concentrations of KKT tablets for intervention. The concentrations of 10ug / ml (KKT-L+A / PR / 8), 20ug / ml (KKT-M+A / PR / 8), and 40ug / ml (KKT-H+A / PR / 8) were selected for plaque assay to further confirm the antiviral efficacy of KKT tablets. Figure 3 As shown, the results show that Keke tablets have good efficacy in inhibiting influenza virus replication.
[0034] Trial on the efficacy of 3g cough tablets in treating cough after influenza virus infection
[0035] To further confirm the efficacy of Keke tablets in treating cough after influenza virus infection, 48 Hartley female guinea pigs were selected and randomly divided into seven groups after 3 days of adaptive feeding: blank control group, model group, positive drug ASM group (ASM, dose of 21.62 mg / kg), Keke tablet low-dose group (dose of 41.85 mg / kg), Keke tablet medium-dose group (dose of 83.7 mg / kg), Keke tablet high-dose group (dose of 167.4 mg / kg), 8 in each group. After grinding the drug, the drug suspension was prepared with 0.5% CMC-Na solution for use. Guinea pigs in all groups except the blank control group were infected with 2LD50A / PR / 8 virus drops to construct a cough model after influenza virus infection. The body weight of the guinea pigs was recorded within 2 hours after infection and the drug intervention was carried out for 4 consecutive days. After the last drug administration, the guinea pigs were placed in a WBP plethysmograph and a 0.4 M citric acid solution was nebulized and inhaled for 6 minutes. The waveform was analyzed according to the respiratory signal detected by the instrument, and waveforms ≥ 20 were recorded as one cough. The number of coughs of the guinea pigs within 6 minutes was calculated. Figure 4 and 5 As shown, the analysis results show that Keke tablets have a good effect in inhibiting cough after guinea pig infection. In the figure, control / ctrl is a blank control group, A / PR / 8 is a model group, ASM+A / PR / 8 is a positive drug Asmi group, KKT-L+A / PR / 8 Keke tablets low-dose group, KKT-M+A / PR / 8 Keke tablets medium-dose group, and KKT-H+A / PR / 8 Keke tablets high-dose group.
[0036] Experimental 4-gram cough tablets: basic analysis of drug efficacy
[0037] At the same time, in order to further analyze the pharmacodynamic basis of the present invention, 500 μL serum of guinea pigs was collected, 1 ml of methanol was added to precipitate protein, vortexed to mix, centrifuged at 10000 rpm for 10 min at 4°C, the supernatant was taken, and nitrogen was blown to dryness at room temperature, 100 μL of methanol was re-dissolved, vortexed and ultrasonicated, and centrifuged at 10000 rpm for 10 min at 4°C, the supernatant was taken, and UPLC-Q-Orbitrap HRMS analysis was performed to confirm the chemical composition of the Keke tablet sample through retention time, accurate mass number, secondary mass spectrometry and other information. Since no component was detected in the serum of the model group (blank), the result of detection in the drug-administered serum was judged to be the drug migration component in the serum. Figure 6 As shown in the figure, the results show that the main material basis of Keke tablets in treating post-infectious cough is ephedrine, golden corydaline, isophoroside, papaverine and other substances. The network pharmacology analysis of its main material basis and post-infectious cough was performed, such as Figure 7 As shown, the analysis results show that Keke tablets can treat post-infectious cough from multiple levels.
[0038] Experimental study on the efficacy of 5g cough tablets in treating airway and lung tissue damage after influenza virus infection
[0039] To further confirm the efficacy of the present invention, BALF fluid and bronchial and lung samples of guinea pigs were collected for further analysis. The results showed that Keke Tablets have the effects of inhibiting the enrichment of guinea pig airway immune cells, improving bronchial and lung tissue inflammatory infiltration, and inhibiting the influenza virus load in lung tissue, thereby playing the role of treating cough after influenza virus infection. Figure 8 As shown in the figure, the effect of Keke tablets on respiratory pathology after guinea pig influenza virus infection was tested. The results showed that Keke tablets have a good effect in improving airway and lung tissue damage after guinea pig influenza virus infection. Fig. 9 As shown in the figure, the effect of Keke tablets on the weight, lung index, bronchial index and spleen index of guinea pigs infected with influenza virus was tested. The results showed that Keke tablets had a good improvement effect on the weight loss, lung tissue and bronchial damage and inflammation of guinea pigs caused by influenza virus infection, and the improvement effect was better than that of the positive drug Asmi. Fig.10 and 11 As shown in the figure, the effect of Keke tablets on the enrichment of immune cells and the secretion of proinflammatory factors in the lung tissue of guinea pigs after influenza virus infection was tested. The results showed that Keke tablets had a good regulatory effect on the classification and enrichment of immune cells in the bronchoalveolar lavage fluid of guinea pigs caused by influenza virus infection, and at the same time, it could effectively improve the secretion of proinflammatory factors in the lung tissue, and the improvement effect was comparable to that of the positive drug Asami. Fig.12 and 13 As shown, the effect of Keke tablets on the viral load of lung tissue in guinea pigs after influenza virus infection was tested. The results showed that Keke tablets had a good inhibitory effect on the virus in the lung tissue of guinea pigs caused by influenza virus infection, and its inhibitory effect was better than that of the positive drug group.
[0040] Finally, it should be emphasized that the above is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.
Claims
1. The use of a cough-killing preparation in the preparation of a medicine for treating cough after influenza virus infection, characterized in that The cough suppressant preparation comprises the following components in parts by weight: 360 parts of ephedra, 360 parts of poppy shells, 360 parts of liquorice, 360 parts of bitter almonds, 112.5 parts of radish seeds, 112.5 parts of platycodon grandiflorum and 112.5 parts of gypsum.
2. The use of a Keke preparation in the preparation of a drug for treating airway or lung tissue damage after influenza virus infection, characterized in that The cough suppressant preparation comprises the following components in parts by weight: 360 parts of ephedra, 360 parts of poppy shells, 360 parts of liquorice, 360 parts of bitter almonds, 112.5 parts of radish seeds, 112.5 parts of platycodon grandiflorum and 112.5 parts of gypsum.
3. The use according to claim 1 or 2, characterized in that The cough suppressing preparation is an oral dosage form.
4. The use according to claim 1 or 2, characterized in that The oral dosage form is tablets, granules, powders, pastes, capsules, pills or oral liquid preparations.
5. The use according to claim 1 or 2, characterized in that The cough suppressing preparation is a tablet. The preparation method of the cough suppressing preparation is as follows: 360 parts of ephedra, 360 parts of poppy shell, 360 parts of liquorice, 360 parts of bitter almond, 112.5 parts of radish seeds, 112.5 parts of platycodon and 112.5 parts of gypsum are taken, and the ephedra and poppy shell powder are crushed into fine powder respectively, sieved, and 165 parts of each fine powder are retained for standby use; the remaining coarse powder of ephedra and poppy shell is adjusted to pH 4.5-5.5 with an acidic solution and decocted twice, each time 2 hours, filter, adjust the pH value of the filtrate to 6.5-7.5 with alkaline solution, and keep the medicinal solution for later use; add liquorice, bitter almond, radish seed, platycodon and gypsum into water and boil twice, each time for 1 hour, filter, combine the filtrate with the above solution, and concentrate under reduced pressure to a clear paste with a relative density of 1.20-1.26, add fine powder of ephedra and poppy shell, mix well, dry, crush into fine powder, add auxiliary materials, mix well, granulate, dry, press into tablets, and wrap with film coating.