A traditional Chinese medicine composition for treating cough of wind-phlegm lingering in the lung

Abstract

The present application relates to a kind of traditional Chinese medicine composition for treating wind phlegm type cough of lung, the pharmaceutical composition is composed of effective component and medically acceptable auxiliary materials, wherein the effective component is made of the following weight percentage of raw drug: jujube 12~18%, purple yam 12~18%, winter flower 12~18%, dried ginger 6~9%, mango kernel 20~28%, schizonepeta 12~18%, liquorice 6~9%.The effect of the pharmaceutical composition for treating wind phlegm type cough of lung is remarkable.

Description

Technical Field

[0001] This invention relates to medical preparations, specifically to pharmaceutical preparations containing undetermined structures derived from traditional Chinese medicine, which are suitable for treating cough of the wind-phlegm-lung type. Background Technology

[0002] Cough is one of the most common complaints among patients visiting the respiratory department. It is a normal protective response mechanism of the human body that helps clear secretions and foreign objects from the respiratory tract. According to the Chinese Medical Association's "Guidelines for the Diagnosis and Treatment of Cough (2015 Edition)," cough is usually classified into three categories based on the course of the disease: acute cough (<3 weeks), subacute cough (3-8 weeks), and chronic cough (>8 weeks).

[0003] Current epidemiological surveys both domestically and internationally show that the incidence of cough in the general population is over 10%, with approximately one-third of patients experiencing cough symptoms for more than three weeks. Subacute and chronic coughs account for a high proportion. This is particularly true in children, whose respiratory and immune systems are not yet fully developed, resulting in a persistently high incidence of chronic cough. In developed countries such as Europe and the United States, the prevalence of chronic cough in children can be as high as 22%. In my country, cough is the most common chief complaint and symptom in pediatric respiratory clinics. Statistics show that 70%-80% of children visiting the clinic daily present with fever accompanied by cough or simply with a cough.

[0004] Although this disease is often self-limiting, some patients experience persistent symptoms, and the longer the course of the disease, the greater the impact on their quality of life. For children, prolonged and severe coughing not only affects sleep, learning, and growth and development, but may also lead to a series of complications such as developmental delays, learning disabilities, insomnia, anxiety, and bedwetting, placing a heavy burden on families and society.

[0005] Currently, modern medicine treats cough primarily by addressing the underlying cause, followed by the use of antitussives and expectorants. Antitussives can be broadly categorized into central and peripheral types. Central antitussives include codeine, pholcodone, and dextromethorphan, while peripheral antitussives include noscapine and phenpropheniramine. Expectorants include ambroxol, guaiacol glycerol ether, and myrtle oil. Nebulized inhalation of corticosteroids, leukotriene receptor antagonists, or bronchodilators can also help reduce inflammation and suppress cough. It is important to note that medication use is more limited in children. Expectorants should be used with caution in children under 2 years of age, and central antitussives should also be used cautiously and only after thorough evaluation in children with severe coughs.

[0006] The "Guidelines for the Diagnosis and Management of Cough at the Primary Level in China (2024)" points out that Traditional Chinese Medicine (TCM) is one of the important means of treating cough. Especially in children with chronic cough, oral administration of TCM based on syndrome differentiation is a common treatment method. The "Expert Consensus on the Diagnosis and Treatment of Post-Infectious Cough in Children with Integrated Traditional Chinese and Western Medicine (2024 Edition)" proposes that "wind-evil invading the lungs, causing the lungs to lose their ability to disperse and purify, resulting in the accumulation of fluids into phlegm, and wind and phlegm binding and stagnating in the lungs" is one of the pathogenesis of cough. At the same time, the consensus classifies the TCM syndromes of children's cough into wind-phlegm lingering in the lungs, phlegm-heat retention, and liver fire invading the lungs, and recommends Zhike San combined with Er Chen Tang with modifications as a treatment for wind-phlegm lingering in the lungs syndrome.

[0007] However, the above prescriptions still have some shortcomings. The combination of Zhike San and Er Chen Tang recommended in the consensus is composed of "Platycodon grandiflorus, Schizonepeta tenuifolia, Aster tataricus, Stemona japonica, Cynanchum paniculatum, Citrus reticulata peel, Pinellia ternata, Poria cocos, Glycyrrhiza uralensis, Zingiber officinale, and Prunus mume". The ingredients are complicated and the proportion of expectorants is relatively large. It is more suitable for cough with phlegm-dampness syndrome, but it is not strong enough to warm the lungs and dispel wind. It does not fully meet the pathogenesis characteristics of cough with wind-phlegm lingering in the lungs in children.

[0008] Furthermore, patent application CN 104721798 A discloses a traditional Chinese medicine composition for relieving cough. This composition is made from 3-20 parts by weight of Aster tataricus, 3-20 parts by weight of Tussilago farfara, 3-20 parts by weight of dried ginger, 5-60 parts by weight of mango kernel, 3-20 parts by weight of Schizonepeta tenuifolia, and 3-20 parts by weight of Belamcanda chinensis. Although the above composition can treat acute and subacute cough and has the effect of warming the lungs, resolving phlegm, and relieving cough, its efficacy in dispelling wind and relieving cough is insufficient.

[0009] Based on clinical observations, the inventors have found that many children with coughs often exhibit an irritating cough, sometimes accompanied by a small amount of thin, white sputum, in the later stages of their illness. This cough is frequently triggered or aggravated by exposure to cold or wind. The inventors believe this type of cough falls under the category of "wind-phlegm lingering in the lungs syndrome." This syndrome arises from external wind pathogens lingering in the lungs, causing impaired lung qi circulation, resulting in frequent and persistent coughs. The obstructed lung qi leads to the condensation of fluids in the upper jiao (upper burner) into phlegm, which is then expectorated as thin, white sputum. With prolonged illness, the wind pathogens and the phlegm in the lungs combine, further obstructing the defensive qi. Exposure to wind and cold triggers the internal pathogens, leading to recurrent coughs. Treatment for this condition should begin with warming the lungs, resolving phlegm, dispelling wind, and stopping the cough. Considering the physiological and pathological characteristics of children, the medication should not be too strong; it should warm the lungs and resolve phlegm without causing dryness or heat, and promote lung qi circulation and dispel wind without depleting qi. Summary of the Invention

[0010] The technical problem to be solved by the present invention is to provide a pharmaceutical composition for treating cough with phlegm lingering in the lungs. The pharmaceutical composition is simple and effective, and has a significant effect on treating cough with phlegm lingering in the lungs.

[0011] The technical solution of the present invention to solve the above problems is as follows:

[0012] A pharmaceutical composition for treating cough due to phlegm retention in the lungs, the pharmaceutical composition comprising an active ingredient and medically acceptable excipients, wherein the active ingredient is made from the following raw material in weight percentages:

[0013] Platycodon grandiflorus 12-18%, Aster tataricus 12-18%, Tussilago farfara 12-18%, Zingiber officinale 6-9%, Mango kernel 20-28%, Schizonepeta tenuifolia 12-18%, Glycyrrhiza uralensis 6-9%.

[0014] The optimal proportions of the active ingredients in the above pharmaceutical composition are derived from the following raw materials by weight percentage:

[0015] Platycodon grandiflorus 15.4%, Aster tataricus 15.4%, Tussilago farfara 15.4%, Zingiber officinale 7.7%, Mango kernel 23.0%, Schizonepeta tenuifolia 15.4%, Glycyrrhiza uralensis 7.7%.

[0016] The active ingredient of the above-mentioned pharmaceutical composition is obtained by the following method:

[0017] (1) Take the raw material and add 12 times the amount of water to decoct for 2 hours, filter, add 10 times the amount of water to the filter residue and decoct for 2 hours, filter, combine the two decoctions, and concentrate under reduced pressure to a solution with a relative density of 1 g / mL at 25°C.

[0018] (2) Take the solution obtained in step (1), add flavoring agent, sterilize, and the effective ingredients are obtained.

[0019] The pharmaceutical composition described in this invention is a conventional compound, granule, tablet or capsule.

[0020] This drug combination consists of seven ingredients: Platycodon grandiflorus, Aster tataricus, Coltsfoot flower, Dried ginger, Mango kernel, Schizonepeta tenuifolia, and Licorice. Among them, Aster tataricus has a strong ability to eliminate phlegm and stop cough, mainly by moistening the lungs and lowering lung qi. Platycodon grandiflorus is good at clearing the lungs and eliminating phlegm, mainly by clearing the lungs and dispelling pathogens. The two are used together as the chief ingredient, one lowering and the other ascending, which can clear the lungs and eliminate phlegm, and also lower qi and stop cough. Dried ginger is good at warming and transforming cold phlegm, and can warm the spleen yang to eliminate the source of phlegm. Coltsfoot flower eliminates phlegm and stops cough, and can moisten the lungs and expel phlegm. The two are used together as the assistant ingredient. Schizonepeta tenuifolia dispels wind and releases the exterior, and helps the chief ingredient to disperse residual pathogens in the lungs. Mango kernel clears heat and eliminates stagnation, and can help the assistant ingredient to transform phlegm in the middle jiao. The two are used together as the adjuvant ingredient. Licorice is the guiding ingredient. It can not only harmonize the various ingredients, but also support the efficacy of the medicine in the middle and upper jiao, and at the same time, it can also moderate the pungent and hot nature of dried ginger. This formula combines lung-clearing and lung-soothing effects, eliminating pathogenic factors without harming the body's vital energy, and warming the lungs without causing dryness or heat. It can resolve phlegm and stop coughing, clear away pathogenic factors and harmonize lung qi, thus achieving the combined effects of warming the lungs, dispelling wind, resolving phlegm, and stopping coughing.

[0021] The formulation of the pharmaceutical composition according to the present invention is improved from the formulation of the patent application with publication number CN 104721798 A, and the improvements lie in: 1. In terms of medicinal ingredients, Belamcandae Rhizoma is omitted, and Platycodi Radix and Glycyrrhizae Radix et Rhizoma are added; 2. In terms of prescription structure, the newly added Platycodi Radix is combined with Asteris Radix et Rhizoma as the monarch drug, and the original monarch drug Farfarae Flos is combined with Zingiberis Rhizoma as the minister drug. The above improvements can produce the following beneficial effects: First, Platycodi Radix has a light and ascending property, so it disperses the lung without purging the lung; Asteris Radix et Rhizoma is warm but not hot, and moistens the lung without dryness. The combination of the two as monarch drugs ensures proper dispersion and descent without damaging qi and yin, which conforms to the physiological characteristics of children with delicate zang-fu organs that are prone to cold and heat, and can better eliminate pathogenic qi in the lung; Second, the added Glycyrrhizae Radix et Rhizoma that moistens the lung and harmonizes medicinals can buffer the warm and dry property of medicinals such as Zingiberis Rhizoma, so as to enhance the antitussive effect of the entire pharmaceutical composition.

[0022] It can be seen from the above analysis that the therapeutic method of the prescription of the present invention is changed from the original "warming the lung to dispel cold, resolving phlegm to stop cough" to "warming the lung to resolve phlegm, dispelling wind to stop cough", which enhances the effect of dispelling wind to stop cough, and can significantly improve the therapeutic effect on cough of wind-phlegm lingering in the lung type.

[0023] Pharmacodynamic Study (Animal Experiment)

[0024] 1 Drugs and Reagents:

[0025] Blank group: Cream base, purchased from Sigma-Aldrich (Shanghai) Trading Co., Ltd.

[0026] Patent formulation group: The formulation of Example 1 described below is a mixture.

[0027] Positive drug control group: Compound Methoxyphenamine Capsules, produced by Changxing Pharmaceutical Co., Ltd., with the approval number H20020393.

[0028] Control formulation group: The patented formulation of the patent publication No. CN 104721798 A (the specific dosage of the six medicinal ingredients is: 10 g of Asteris Radix et Rhizoma, 10 g of Farfarae Flos, 10 g of Zingiberis Rhizoma, 15 g of Mangiferae Semen, 10 g of Schizonepetae Spica, 10 g of Belamcandae Rhizoma), which is prepared into a mixture according to the process in Example 1 thereof.

[0029] Experimental animals: 30 SPF-grade C57BL / 6 mice, female, 6 weeks old, purchased from Guangdong Vital River Laboratory Animal Technology Co., Ltd. [SCXK (Guangdong) 2022-0063].

[0030] Main reagents and materials: Baisha tobacco (produced by Hunan Tobacco Industry Co., Ltd., tar content 10 mg, nicotine content 0.8 mg, carbon monoxide content 12 mg); polysorbate-80, LPS, capsaicin (natural) (Shanghai Yuanye Biotechnology Co., Ltd., S24644, S11060, B20693); D12492 high-fat diet (Research Diets, Inc., 60 kcal% Fat); IL-1β, CXCL1 / KC, TNF-α ELISA kits (China Lianke Biotechnology, EK201B, EK296, EK282); isopropanol, anhydrous ethanol (Sinopharm Chemical Reagent Co., Ltd., 80109218, 10009218).

[0031] 2 Experimental Methods

[0032] 2.1 Modeling of cough due to wind-phlegm lingering in the lungs

[0033] A mouse model of post-infectious cough with phlegm was established using a combination of fumigation, LPS nasal drops, capsaicin nebulization, wind stimulation, a high-fat diet, and cold stimulation with ice water. The modeling period was 14 days. The control group was fed routinely throughout the modeling process and did not receive the above stimulation. Fumigation stimulation was administered on days 1-7. Mice were placed in a self-made fumigation device and exposed to the smoke produced by 10 cigarettes of Baisha tobacco for 30 minutes each time, once daily for 7 consecutive days. LPS nasal drops were administered on days 8, 11, and 14. Mice were lightly anesthetized with isoflurane and then given LPS-PBS working solution via nasal drops. The concentration of the LPS working solution was 0.4 mg / mL, and the administration volume was calculated at 1 μL / g body weight, i.e., the total amount of nasal drops per mouse was determined based on the body weight on the day of administration. Capsaicin nebulization cough was induced on days 9, 10, 12, and 13. Mice were placed in a transparent nebulization chamber to receive capsaicin nebulization stimulation. The concentration of the capsaicin working solution was 1×10⁻⁶. -4 mol / L, 3 min each time, once daily. After nebulization, observe changes in respiration, sneezing, face scratching, oral and nasal secretions, and activity status of mice. On days 1, 3, 5, 7, 10, and 13, administer wind stimulation. Place mice in a ventilated cage and apply wind stimulation using an electric fan at a speed of approximately 5 m / s. The ambient temperature is (24±2)°C, and the relative humidity is 50%-55%, for 1.5 h each time, to simulate wind-induced cough and its aggravation upon exposure to wind.

[0034] From day 1 to day 14 of modeling, the patient was fed a high-fat diet continuously. On days 9, 10, 12, and 13, the patient was given continuous ice water for 8 hours of cold stimulation to help establish the manifestations of wind-phlegm retention in the lungs. The final modeling treatment was completed on the morning of day 14, and the model was evaluated in the afternoon.

[0035] 2.2 Animal grouping and administration methods

[0036] After 3 days of acclimatization, mice were divided into four groups according to the experimental design: blank group, model group, positive drug control group, control group, and patented formula group, with 6 mice in each group. Except for the blank group, all other groups underwent a combined model of post-infectious cough with phlegm lingering in the lungs.

[0037] Dosage calculation: Referring to the section on animal dosage and equivalent dose in *Methodology of Pharmacological Research of Traditional Chinese Medicine* (People's Medical Publishing House, 3rd edition, 2011, pp. 1261-1264), the equivalent dose for mice was determined to be approximately 12 times that for humans. The dosage for the experimental and control groups was 12g crude drug / kg; the positive control group was given compound methoxyphenamine capsules at a dosage of 1.2 capsules / kg. After model establishment and evaluation, gavage administration began once daily for 7 consecutive days, with the gavage volume calculated as 0.12 mL / 10g body weight. The blank group and model group were given an equal volume of physiological saline, the positive control group was given compound methoxyphenamine capsule suspension, and the control group and patent formula group were given the corresponding traditional Chinese medicine solutions.

[0038] Preparation of the patented drug formulation: Extracted according to the method in Example 1, and concentrated to a solution of 1.0 g / mL;

[0039] Preparation of drugs for the control group: The drug solutions used in the experimental and control groups were prepared by the following method: Take the drug, add 12 times the amount of water and decoct for 2 hours, filter, add 10 times the amount of water to the residue and decoct for 2 hours, filter, combine the decoctions, and concentrate under reduced pressure to a solution with a relative density of 1.0 g / mL at 25℃.

[0040] 2.3 Sample Collection

[0041] Samples were collected after the last administration. Mice were anesthetized by intraperitoneal injection of sodium pentobarbital. After adequate anesthesia, blood was collected from the eyeballs. The blood samples were allowed to stand for 2 hours before centrifugation to separate the serum for subsequent detection of inflammatory factors. After blood collection, lung tissue was rapidly separated. The left lung was used to calculate the wet / dry weight ratio; the upper lobe of the right lung was fixed in 4% paraformaldehyde for subsequent HE staining and pathological evaluation; the remaining lung tissue was flash-frozen in liquid nitrogen and stored at -80°C.

[0042] 2.4 General condition and weight observation

[0043] During the experiment, the mice were observed daily for their mental state, activity level, coat luster, food and water intake, fecal characteristics, respiratory status, and mortality. Body weight was measured at baseline, at the end of modeling, and after the last drug administration. The rate of change during the modeling period, the rate of change during the drug administration period, and the overall rate of change were calculated to evaluate the effects of modeling and drug intervention on the overall condition of the mice.

[0044] 2.5 Post-infectious cough disease score

[0045] Post-infectious cough (PIC) behavioral evaluation was conducted after modeling and the last administration. The main symptom indicators of PIC cough were the number of coughs within 3 minutes after capsaicin nebulization-induced cough, cough latency, paroxysmal cough, choking or sneezing, and neck extension or shortness of breath-like movements. Each item was scored semi-quantitatively from 0 to 3 points, and the sum of the five items constituted the PIC cough symptom score, ranging from 0 to 15 points. An increase in the number of coughs within 3 minutes after induction, a shortened cough latency, and an elevated PIC cough symptom score indicated a worsening of post-infectious cough symptoms.

[0046] 2.5 Evaluation of the Wind-Phlegm-Lung Syndrome Model

[0047] The evaluation of the Wind-Phlegm-Lingering Lung syndrome includes two dimensions: Wind-Evil Latent in the Lung and Wind-Phlegm-Lingering. Manifestations of Wind-Evil Latent in the Lung include increased cough sensitivity after induction, itchy throat / immediate cough upon stimulation, and worsening of symptoms after exposure to wind, each scored from 0 to 3. Manifestations of Wind-Phlegm-Lingering include changes in body shape or weight, mental state, hair luster, activity level, behavioral response, food and water intake, diarrhea, and the color of the lips, ears, and nails, recorded on a 0 / 1 scale, with the number of positive items counted. The Wind-Phlegm-Lingering Lung syndrome score is the sum of the Wind-Evil Latent in the Lung score and the number of positive items in Wind-Phlegm-Lingering; a higher score indicates a more severe Wind-Phlegm-Lingering Lung syndrome.

[0048] 2.7 Lung W / D Measurement

[0049] Take tissue from the left lung, remove surface blood and fluid, and immediately weigh the lung wet weight (W). Then bake and dry it in an 80°C oven until constant weight. Take it out and weigh it again, record the dry weight (D), and calculate the lung wet / dry weight ratio (W / D).

[0050] 2.8 HE staining and pathological evaluation of lung tissue

[0051] Tissue from the right upper lobe of the lung, fixed in 4% paraformaldehyde, was routinely dehydrated, embedded in paraffin, sectioned, and stained with hematoxylin and eosin (HE). The sections were then scanned using a whole-section scanning system for digital pathological analysis. By combining low-power whole-section structure and multiple high-power representative fields, a semi-quantitative evaluation was performed on alveolar structural destruction, alveolar septal thickening, inflammatory cell infiltration, localized consolidation or exudation, and peri-airway / vascular inflammatory cell aggregation.

[0052] The HE score includes the ATS lung injury score and the airway / perivascular inflammation score. The ATS lung injury score reflects changes such as inflammatory cells, hyaline membranes, protein-like exudates, and thickening of alveolar septa in the alveolar space and interstitium; the airway / perivascular inflammation score is scored from 0 to 4 to evaluate the degree of perivascular infiltration of inflammatory cells.

[0053] 2.9 PAS staining and evaluation of airway mucus secretion

[0054] Paraffin sections of lung tissue were stained with periodic acid-Schiff (PAS) to observe airway epithelial mucus secretion and goblet cell proliferation. PAS-stained sections were analyzed using digital image analysis to identify effective tissue areas and purplish-red PAS-positive signals, and the proportion of PAS-positive areas was calculated. To facilitate inter-group comparisons, the proportion of PAS-positive areas was converted to a 0-4 score (PAS conversion score = PAS-positive area proportion / 2.5).

[0055] 2.10 Serum inflammatory factor detection

[0056] After the last administration, blood was collected from the eyes of mice in each group. After standing for 2 hours, serum was separated by centrifugation at 4°C and stored at -20°C. Serum levels of IL-1β, TNF-α, and CXCL1 / KC were detected using IL-1β, TNF-α, and CXCL1 / KC kits, strictly following the kit instructions.

[0057] 2.11 Statistical Methods

[0058] Quantitative data are expressed as mean ± standard deviation (SD). Two-tailed Welch t-tests were used for comparisons between groups, without correction for multiple comparisons.

[0059] 3 Results

[0060] 3.1 General condition and weight changes of mice in each group

[0061] During the experiment, the mice in the control group maintained stable mental state, activity level, food and water intake, and coat condition. There were no statistically significant differences in baseline body weight among the groups. After modeling, the body weight or rate of change during the modeling period decreased in the model group, positive control group, control group, and patented formulation group compared to the control group. After the last administration, there were no statistically significant differences in body weight among the groups, indicating that the overall body weight of each group tended to recover after intervention (see Table 1).

[0062] Table 1 Comparison of body weight and body weight change rate among different groups of mice

[0063] index Blank group Model group Positive drug control group control group Patent Group Baseline body weight (g) 20.88±0.51 21.77±1.17 22.08±1.49 21.72±0.88 21.45±0.90 Post-modeling body weight (g) 23.70±0.80 <![CDATA[22.03±0.54 ## ]]> <![CDATA[20.82±0.25 ### ]]> <![CDATA[22.47±0.96 # ]]> <![CDATA[21.17±2.14 # ]]> Last weight (g) 23.52±0.26 23.58±0.69 23.23±1.95 23.13±1.05 23.08±1.18

[0064] Note: Data are expressed as mean ± SD, and n=6 for each group. # , ## , ### These represent P<0.05, P<0.01, and P<0.001 compared to the control group, respectively.

[0065] 3.2 Coughing behavior and lung-wind syndrome scores of mice in each group after infection

[0066] At T1, compared with the control group, the number of coughs within 3 minutes was significantly increased, the cough latency was significantly shortened, and the PIC cough symptom score and the wind-phlegm-lung syndrome score were significantly increased in all groups, indicating that the mouse model of post-infectious cough with wind-phlegm-lung syndrome was successfully established. Before entering the drug administration phase, each model group had formed relatively consistent disease and syndrome manifestations (see Table 2).

[0067] At the time of the last administration (T2), the model group maintained a high cough response and syndrome score. Compared with the model group, the cough latency was significantly prolonged in the positive drug control group, the control group, and the patent formula group, and the PIC cough symptom score and the wind-phlegm-lung syndrome score were significantly reduced. Among them, the number of coughs in the patent formula group was significantly lower than that in the model group (P=0.0170), and the number of coughs in the 3 minutes, the cough latency, the PIC cough symptom score, and the wind-phlegm-lung syndrome score were all improved compared with the control group. The difference in the wind-phlegm-lung syndrome score between the groups was statistically significant (P<0.0001).

[0068] Table 2 Comparison of cough behavior indicators and lung-wind syndrome scores in mice after infection among different groups

[0069] time index Blank group Model group Positive drug control group control group Patent Group T1 Number of coughs (3 minutes) 4.83±2.86 <![CDATA[21.33±10.09 ## ]]> <![CDATA[19.83±7.99 ## ]]> <![CDATA[17.33±8.45 # ]]> <![CDATA[18.33±10.01 # ]]> T1 Cough incubation period (s) 111.83±27.45 <![CDATA[19.50±12.91 ### ]]> <![CDATA[31.67±10.01 ### ]]> <![CDATA[27.67±11.25 ### ]]> <![CDATA[34.50±8.38 ### ]]> T1 PIC Cough Symptomatic Score 1.00±1.10 <![CDATA[11.67±1.21 ### ]]> <![CDATA[10.67±1.21 ### ]]> <![CDATA[10.50±1.38 ### ]]> <![CDATA[10.83±0.75 ### ]]> T1 Wind-phlegm lingering in the lungs syndrome scoring 2.33±1.63 <![CDATA[11.50±2.43 ### ]]> <![CDATA[12.17±2.14 ### ]]> <![CDATA[12.50±0.55 ### ]]> <![CDATA[12.83±0.75 ### ]]> T2 Number of coughs (3 minutes) 3.50±3.15 <![CDATA[14.67±7.61 # ]]> <![CDATA[6.83±1.17 # ]]> <![CDATA[9.33±1.86 ## ]]> <![CDATA[3.83±1.17 *▲▲▲ ]]> T2 Cough incubation period (s) 122.17±16.15 <![CDATA[20.00±8.83 ### ]]> <![CDATA[89.67±10.17 ##*** ]]> <![CDATA[65.50±9.12 ###*** ]]> <![CDATA[121.67±8.96 ***▲▲▲ ]]> T2 PIC Cough Symptomatic Score 0.50±0.55 <![CDATA[11.00±1.41 ### ]]> <![CDATA[4.17±0.75 ###*** ]]> <![CDATA[5.83±0.75 ###*** ]]> <![CDATA[3.17±0.75 ###***▲▲▲ ]]> T2 Wind-phlegm lingering in the lungs syndrome scoring 1.33±1.03 <![CDATA[12.50±1.22 ### ]]> <![CDATA[6.00±0.63 ###*** ]]> <![CDATA[7.17±0.75 ###*** ]]> <![CDATA[4.17±0.75 ###***▲▲▲ ]]>

[0070] Note: Data are expressed as mean ± SD, and n=6 for each group. T1 is the time from the end of modeling to before drug administration; T2 is the time from the last drug administration. # , ## , ### These represent P<0.05, P<0.01, and P<0.001 compared to the control group, respectively. * , ** , *** These represent P<0.05, P<0.01, and P<0.001 compared to the model group, respectively. ▲ , ▲▲ , ▲▲▲ The values ​​represent P<0.05, P<0.01, and P<0.001, respectively, when comparing the patent formulation with the control formulation.

[0071] 3.3 Lung wet / dry weight and wet / dry ratio of mice in each group

[0072] Compared with the blank group (see Table 3), the lung wet weight and lung dry weight of the model group were increased, indicating that the lung tissue weight and inflammation-related changes were increased in the model group. Compared with the model group, the lung wet weight and wet / dry ratio of the positive drug control group, the control group, and the patented formula group were all decreased; among them, the lung wet weight and wet / dry ratio of the patented formula group decreased more significantly and were both lower than those of the control group (all P<0.0001).

[0073] Table 3 Comparison of wet and dry lung weight and wet / dry ratio in mice of different groups

[0074] index Blank group Model group Positive drug control group control group Patent Group Lung wet weight (g) 0.067±0.005 <![CDATA[0.075±0.005 # ]]> <![CDATA[0.065±0.001 ** ]]> <![CDATA[0.063±0.001 ** ]]> <![CDATA[0.056±0.001 ##***▲▲▲ ]]> Lung dry weight (g) 0.013±0.001 <![CDATA[0.015±0.002 # ]]> <![CDATA[0.014±0.001 ### ]]> <![CDATA[0.014±0.001 ### ]]> <![CDATA[0.014±0.000 ##▲ ]]> Wet / dry ratio 5.31±0.29 4.94±0.35 <![CDATA[4.49±0.06 ###* ]]> <![CDATA[4.39±0.05 ###* ]]> <![CDATA[3.99±0.05 ###***▲▲▲ ]]>

[0075] Note: Data are expressed as mean ± SD, with n=6 for each group. The wet / dry ratio is the ratio of lung wet weight to lung dry weight. # , ## , ### These represent P<0.05, P<0.01, and P<0.001 compared to the control group, respectively. * , ** , *** These represent P<0.05, P<0.01, and P<0.001 compared to the model group, respectively. ▲ , ▲▲ , ▲▲▲ The values ​​represent P<0.05, P<0.01, and P<0.001, respectively, when comparing the patent formulation with the control formulation.

[0076] 3.4 HE staining pathological results of lung tissue from each group of mice

[0077] HE staining results showed that ( Figure 1 In the blank control group, the alveolar structure was clear, the alveolar septa were thin, and no extensive consolidation or significant perivascular inflammation was observed. The model group showed more severe lung lesions, mainly manifested as thickened alveolar septa, localized alveolar cavity compression or structural disorder, increased inflammatory cell infiltration, and peri-aqueous inflammatory cell aggregation in the airways or blood vessels. Compared with the model group, the lung lesions in the positive control group, the control group, and the patented formula group were all reduced to varying degrees.

[0078] Semi-quantitative scoring (see Table 4) showed that the ATS lung injury score, airway / perivascular inflammation score, and combined HE score in the model group were significantly higher than those in the control group (all P < 0.0001). The scores in all treatment groups were lower than those in the model group. The ATS lung injury score in the patented formula group was lower than that in the control formula group (P < 0.001), and the combined HE score was also lower (P = 0.0364); the airway / perivascular inflammation score showed a decreasing trend compared to the control formula group (P = 0.0846).

[0079] Table 4 Comparison of HE pathological semi-quantitative scores of lung tissue from different groups of mice

[0080] index Blank group Model group Positive drug control group control group Patent Group ATS lung injury score 0.15±0.07 <![CDATA[0.65±0.07 ### ]]> <![CDATA[0.37±0.05 ###*** ]]> <![CDATA[0.49±0.04 ###** ]]> <![CDATA[0.31±0.07 ##***▲▲▲ ]]> Airway / perivascular inflammation score 0.72±0.26 <![CDATA[3.13±0.37 ### ]]> <![CDATA[2.17±0.25 ###*** ]]> <![CDATA[2.61±0.19 ###* ]]> <![CDATA[2.07±0.61 ##** ]]> HE combined score 0.87±0.32 <![CDATA[3.78±0.45 ### ]]> <![CDATA[2.54±0.29 ###*** ]]> <![CDATA[3.10±0.22 ###* ]]> <![CDATA[2.38±0.62 ###**▲ ]]>

[0081] Note: Data are expressed as mean ± SD, and n=6 for each group. The combined HE score is a composite index combining the ATS lung injury score and the airway / perivascular inflammation score. # , ## , ### These represent P<0.05, P<0.01, and P<0.001 compared to the control group, respectively. * , ** , ***These represent P<0.05, P<0.01, and P<0.001 compared to the model group, respectively. ▲ , ▲▲ , ▲▲▲ The values ​​represent P<0.05, P<0.01, and P<0.001, respectively, when comparing the patent formulation with the control formulation.

[0082] 3.5 Results of PAS staining and airway mucus secretion in mice of each group

[0083] PAS staining results showed that ( Figure 2 The PAS positive signal in the lung tissue of the model group was significantly enhanced compared to the control group, indicating an increase in airway mucus secretion and mucus-related positive components after modeling. Quantitative results showed that the proportion of PAS positive area and the PAS conversion score in the model group were significantly higher than those in the control group (both P=0.0033). Compared with the model group, the PAS index was reduced in both the control group and the patented formula group, with a larger decrease in the patented formula group; the proportion of PAS positive area in the patented formula group was lower than that in the control group (P=0.0306), and the PAS conversion score was also lower in the patented formula group (P=0.0310).

[0084] Table 5 Comparison of PAS-positive area ratio and converted score among mice in each group

[0085] index Blank group Model group Positive drug control group control group Patent Group Percentage of PAS-positive areas (%) 2.67±1.08 <![CDATA[6.84±2.14 ## ]]> <![CDATA[5.92±2.33 # ]]> <![CDATA[4.52±0.75 ##* ]]> <![CDATA[2.18±1.94 **▲ ]]> PAS conversion score 1.07±0.43 <![CDATA[2.73±0.85 ## ]]> <![CDATA[2.37±0.93 # ]]> <![CDATA[1.81±0.30 ##* ]]> <![CDATA[0.87±0.78 **▲ ]]>

[0086] Note: Data are expressed as mean ± SD, and n=6 for each group. PAS conversion score = PAS positive area ratio / 2.5, with a maximum score of 4. # , ## , ### These represent P<0.05, P<0.01, and P<0.001 compared to the control group, respectively. * , ** , *** These represent P<0.05, P<0.01, and P<0.001 compared to the model group, respectively. ▲ , ▲▲ , ▲▲▲ The values ​​represent P<0.05, P<0.01, and P<0.001, respectively, when comparing the patent formulation with the control formulation.

[0087] 3.6 Results of serum inflammatory factor detection in each group of mice

[0088] ELISA results (see Table 5) showed that, compared with the blank group, serum IL-1β, TNF-α, and CXCL1 / KC levels in the model group were significantly increased, indicating a significant inflammatory response in mice with post-infection cough and phlegm-related lung syndrome. Compared with the model group, the levels of the three inflammatory factors were decreased in the positive drug control group, the control group, and the patented formula group. The levels of IL-1β, TNF-α, and CXCL1 / KC in the patented formula group were all lower than those in the control group, and the differences were statistically significant, indicating that the patented formula had a more prominent inhibitory effect on the increase of inflammatory factors.

[0089] Table 6 Comparison of serum inflammatory factor levels in mice of different groups

[0090] index Blank group Model group Drug group control group Patent Group IL-1β (pg / mL) 22.82±2.78 <![CDATA[36.18±4.16 ### ]]> <![CDATA[28.11±3.39 #** ]]> <![CDATA[28.60±3.20 ##** ]]> <![CDATA[24.33±0.33 ***▲ ]]> TNF-α (pg / mL) 239.94±27.42 <![CDATA[473.55±45.77 ### ]]> <![CDATA[346.21±33.77 ###*** ]]> <![CDATA[345.50±28.45 ###*** ]]> <![CDATA[311.88±1.63 ##***▲ ]]> CXCL1 / KC (pg / mL) 27.66±3.50 <![CDATA[58.97±6.30 ### ]]> <![CDATA[39.45±4.49 ###*** ]]> <![CDATA[40.50±4.50 ###*** ]]> <![CDATA[34.47±0.50 ##***▲ ]]>

[0091] Note: Data are expressed as mean ± SD, and n=6 for each group. # , ## , ### These represent P<0.05, P<0.01, and P<0.001 compared to the control group, respectively. * , ** , *** These represent P<0.05, P<0.01, and P<0.001 compared to the model group, respectively. ▲ , ▲▲ , ▲▲▲ The values ​​represent P<0.05, P<0.01, and P<0.001, respectively, when comparing the patent formulation with the control formulation.

[0092] 4. Conclusion

[0093] Post-infectious cough is characterized by prolonged course, airway hyperresponsiveness, and persistent inflammation. In Traditional Chinese Medicine (TCM), its pathogenesis can be categorized under external pathogenic factors, wind-dampness lurking in the lungs, and phlegm-dampness obstruction. This study established a mouse model of post-infectious cough with wind-phlegm lingering in the lungs using a combination of factors including fumigation, LPS nasal drops, capsaicin nebulization, wind stimulation, a high-fat diet, and cold water. Results showed that at T1, except for the control group, all groups experienced increased cough frequency and shortened cough latency within 3 minutes, along with elevated PIC cough symptom scores and wind-phlegm lingering in the lungs syndrome scores. Simultaneously, the model group showed elevated lung wet weight, lung dry weight, HE / PAS pathological scores, and serum levels of IL-1β, TNF-α, and CXCL1 / KC. These results confirm the successful establishment of the model from behavioral, symptom, histopathological, and inflammatory factor levels perspectives.

[0094] After intervention with the patented formula, the number of coughs in mice at T2 level decreased, the cough latency period was prolonged, and the PIC cough symptom score and the wind-phlegm-lung syndrome score decreased, indicating that the patented formula can improve post-infectious cough symptoms and alleviate wind-phlegm-lung syndrome. The number of coughs at T2 level, cough latency period, PIC cough symptom score, and wind-phlegm-lung syndrome score in the patented formula group were all improved compared with the control group, indicating that the patented formula is more effective than the patented formula in CN 104721798 B in improving post-infectious cough with wind-phlegm-lung syndrome.

[0095] Lung tissue weighing and morphological results further support the comprehensive intervention effect of the patented formula. The patented formula can reduce lung wet weight and wet / dry ratio, and both lung wet weight and wet / dry ratio are lower than those in the control group. HE score showed that the model group had more severe lung tissue inflammation damage. The patented formula can reduce ATS lung injury score, airway / perivascular inflammation score, and combined HE score, with ATS lung injury score and combined HE score being lower than those in the control group. PAS results showed that the model group had enhanced airway mucus-related positive signal. The patented formula can reduce the proportion of PAS positive area and PAS converted score, both of which are lower than those in the control group, and the differences are statistically significant, indicating that it has a significant ameliorative effect on the increased airway mucus secretion caused by phlegm dampness.

[0096] The results of inflammatory factor analysis showed that IL-1β, TNF-α, and CXCL1 / KC were significantly elevated in the model group. After intervention with the patented formula, all three indicators were lower than those in the model group and lower than those in the control group. CXCL1 / KC is closely related to neutrophil chemotaxis, and IL-1β and TNF-α can amplify local inflammatory responses and promote increased airway sensitivity. The decrease of the above factors showed good biological consistency with prolonged cough latency, reduced syndrome scores, and reduced lung tissue inflammation.

[0097] Compared with the patent formula in CN 104721798 A, the patent formula shows more significant improvements in cough behavior, syndrome score, lung wet / dry weight, HE / PAS pathological indicators, and serum inflammatory factors. The patent formula, based on herbs such as Aster tataricus, Tussilago farfara, dried ginger, and Schizonepeta tenuifolia, is combined with Platycodon grandiflorus, Glycyrrhiza uralensis, and Mango kernel. The formula aims to simultaneously promote lung function, soothe the throat, resolve phlegm, relieve cough, harmonize the properties of other herbs, and improve local inflammatory damage in the lungs. Attached Figure Description

[0098] Figure 1 A panoramic view of HE staining of lung tissue from mice in each group;

[0099] Figure 2 PAS staining was performed on mice in each group. Detailed Implementation

[0100] Example 1 (Mixed Solution)

[0101] 1. Prescription:

[0102] Platycodon grandiflorus 100g, Aster tataricus 100g, Tussilago farfara 100g, Dried ginger 50g, Mango kernel 150g, Schizonepeta tenuifolia 100g, Licorice root 50g.

[0103] 2. Preparation method:

[0104] (1) Take the drug and add 12 times the amount of water to decoct for 2 hours, filter, add 10 times the amount of water to the residue and decoct for 2 hours, filter, combine the decoctions, and concentrate under reduced pressure to a solution with a relative density of 1 g / mL at 25°C.

[0105] (2) Take the solution obtained in step (1), add flavoring agent, sterilize, and dispense into 100mL / bottle.

[0106] Example 2 (Granules)

[0107] 1. Prescription:

[0108] Platycodon grandiflorus 117g, Aster tataricus 113g, Tussilago farfara 100g, Zingiber officinale 47g, Mango kernel 120g, Schizonepeta tenuifolia 100g, Glycyrrhiza uralensis 53g.

[0109] 2. Preparation method:

[0110] (1) Take the medicine and add 12 times the amount of water to decoct for 2 hours, filter, add 10 times the amount of water to the residue and decoct for 2 hours, filter, combine the decoctions, and concentrate under reduced pressure to a thick paste with a relative density of 1.30 at 25°C.

[0111] (2) Take the thick paste obtained in step (1), vacuum dry it to obtain dry paste powder, add excipients to granulate, then dry, granulate and package into 10g / packet.

[0112] Example 3 (Mixed Solution)

[0113] 1. Prescription:

[0114] Platycodon grandiflorus 110g, Aster tataricus 116g, Tussilago farfara 110g, Zingiber officinale 52g, Mango kernel 136g, Schizonepeta tenuifolia 80g, Glycyrrhiza uralensis 46g.

[0115] 2. Preparation method:

[0116] Same as Example 1.

[0117] Example 4 (Granules)

[0118] 1. Prescription:

[0119] Platycodon grandiflorus 90g, Aster tataricus 96g, Tussilago farfara 109g, Dried ginger 58g, Mango kernel 161g, Schizonepeta tenuifolia 96g, Licorice root 40g.

[0120] 2. Preparation method:

[0121] Same as Example 2.

[0122] Example 5 (Mixed Solution)

[0123] 1. Prescription:

[0124] Platycodon grandiflorus 80g, Aster tataricus 92g, Tussilago farfara 112g, Zingiber officinale 59g, Mango kernel 162g, Schizonepeta tenuifolia 99g, Glycyrrhiza uralensis 46g.

[0125] 2. Preparation method:

[0126] Same as Example 1.

[0127] Example 6 (Capsule Formulation)

[0128] The granules prepared according to the prescription in Example 1 were filled into gelatin capsules to form capsules.

[0129] Example 7 (tablet)

[0130] The granules prepared according to the prescription in Example 1 were mixed with sugar powder, and 95% ethanol was used as a wetting agent to form a soft mass. The mass was then granulated through a 16-mesh sieve, dried, and compressed into tablets using conventional methods.

Claims

1. A pharmaceutical composition for treating cough due to phlegm retention in the lungs, the pharmaceutical composition comprising an active ingredient and medically acceptable excipients, wherein said active ingredient is made from the following raw material in weight percentages: Platycodon grandiflorus 12-18%, Aster tataricus 12-18%, Tussilago farfara 12-18%, Zingiber officinale 6-9%, Mango kernel 20-28%, Schizonepeta tenuifolia 12-18%, Glycyrrhiza uralensis 6-9%.

2. The pharmaceutical composition for treating cough due to phlegm retention in the lungs according to claim 1, characterized in that, The active ingredient is made from the following raw materials by weight percentage: Platycodon grandiflorus 15.4%, Aster tataricus 15.4%, Tussilago farfara 15.4%, Zingiber officinale 7.7%, Mango kernel 23.0%, Schizonepeta tenuifolia 15.4%, Glycyrrhiza uralensis 7.7%.

3. A pharmaceutical composition for treating cough of the lung-type with phlegm retention according to claim 1 or 2, characterized in that, The active ingredient is obtained by the following method: (1) Take the raw material and add 12 times the amount of water to decoct for 2 hours, filter, add 10 times the amount of water to the filter residue and decoct for 2 hours, filter, combine the two decoctions, and concentrate under reduced pressure to a solution with a relative density of 1 g / mL at 25°C. (2) Take the solution obtained in step (1), add flavoring agent, sterilize, and the effective ingredients are obtained.

4. The pharmaceutical composition for treating cough due to phlegm retention in the lungs according to claim 3, characterized in that, The traditional Chinese medicine composition is in the form of a mixture, granules, tablets, or capsules.

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