Xiaoqinglong decoction, Xiaoqinglong granules and preparation method thereof

Abstract

The invention discloses a Xiaoqinglong decoction and Xiaoqinglong granules prepared from kadsura longepedunculata, a bronchial asthma model is established by using a mouse, and the pharmacological action of kadsura longepedunculata on treating bronchial asthma is determined to be superior to that of Similarly, a bronchial asthma model is established by a mouse, and the pharmacological action of the Xiaoqinglong decoction and Xiaoqinglong granules obtained after optimization is superior to that of the original formula. The combined optimization method is also suitable for formula optimization of a classic famous formula containing schisandra chinensis and mainly used for treating bronchial asthma.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical preparations, and in particular to a Xiaoqinglong decoction, Xiaoqinglong granules and a preparation method thereof. Background Art

[0002] Bronchial asthma is a common, clinically heterogeneous chronic inflammatory disease of the airways, with a global incidence of about 9% and increasing year by year. Its main characteristics are bronchial inflammation, bronchial wall remodeling, and bronchial hyperresponsiveness to environmental stimuli. The main clinical symptoms are shortness of breath, repeated wheezing, cough, chest tightness, etc. It can also affect diseases such as diabetes, hypertension, coronary heart disease, and chronic kidney disease, seriously affecting the quality of life of patients. In order to avoid the adverse reactions caused by the long-term use of macrolide antibiotics, glucocorticoids and other drugs during its treatment, the use of the traditional Chinese medicine Xiaoqinglong Decoction is considered to be a good strategy for the treatment of bronchial asthma.

[0003] Xiaoqinglong Decoction comes from Zhang Zhongjing's Treatise on Febrile Diseases in the Eastern Han Dynasty. It is composed of eight herbs: ephedra, cinnamon twig, peony root, asarum, dried ginger, schisandra, roasted liquorice, and pinellia. It has the effects of dispelling cold, warming the lungs and eliminating fluid. It is a classic prescription for treating "external cold and internal fluid". Modern pharmacological studies have found that Xiaoqinglong Decoction has anti-allergic, anti-inflammatory, antitussive and antiasthmatic effects. It is mostly used in clinical treatment of respiratory diseases such as chronic bronchitis, bronchial asthma, and chronic obstructive pulmonary disease.

[0004] After combing through the prescription composition of Xiaoqinglong Decoction in the herbal medicines of various dynasties (see Table 1), it was found that all the prescriptions contained Schisandra chinensis (northern Schisandra chinensis). The 2000 edition of the Pharmacopoeia of the People's Republic of China first included Schisandra chinensis (Turcz.) Baill. and Schisandra sphenanthera Rehd. et Wils., which are derived from the Magnoliaceae plant, as two varieties, and called them "Schisandra chinensis" and "Southern Schisandra chinensis" respectively. However, the description of their efficacy is the same as "astringent, invigorating qi and promoting fluid, tonifying the kidney and calming the mind", which has been continued until the current edition of the Pharmacopoeia of the People's Republic of China. The existing Xiaoqinglong Decoction or granules all use Schisandra chinensis, and there is no research and application of Southern Schisandra chinensis in respiratory diseases. Summary of the invention

[0005] In order to solve at least one aspect of the above-mentioned problems and defects existing in the prior art, the present invention provides a Xiaoqinglong decoction, Xiaoqinglong granules and a preparation method thereof, which can prepare a prescription with better efficacy for treating bronchial asthma.

[0006] The first aspect of the present invention provides a Xiaoqinglong decoction, which is prepared with the following Chinese medicinal raw materials: ephedra, cinnamon twig, white peony root, dried ginger, asarum, roasted liquorice, and pinellia, and also includes Schisandra chinensis, with a weight ratio of 2:2:2:2:1:2:3:2.

[0007] The second aspect of the present invention provides a method for preparing Xiaoqinglong granules, wherein the Xiaoqinglong granules are prepared from the following Chinese medicinal raw materials: ephedra, cinnamon twig, white peony root, dried ginger, asarum, roasted liquorice, pinellia tuber, and Schisandra chinensis, with a weight ratio of 2:2:2:2:1:2:3:2;

[0008] The preparation method comprises:

[0009] (1) steam distilling the three medicinal materials of Asarum, Cinnamomum cassia and Schisandra chinensis together to extract a mixture of volatile oils from the three materials, and collecting the aqueous solution after distillation in another device;

[0010] The obtained medicinal residues, the white peony root, ephedra, and licorice root are decocted in water for 2-3 times, each time for 2-4 hours, the decoctions are combined, filtered, the filtrate is combined with the distilled aqueous solution, and concentrated to obtain a first solution;

[0011] The pinellia tuber and dried ginger are crushed into coarse powder, and 60-90% ethanol is used as a solvent. After soaking, the percolation is performed, and the percolation liquid is collected, and the ethanol is recovered and concentrated to a suitable amount, and then combined with the first solution, and the filtrate is allowed to stand, filtered, and the filtrate is concentrated to a suitable amount (e.g., to 1000 mL). After concentration, a second solution is obtained, which is used for standby;

[0012] (2) The second solution is concentrated, spray-dried, an appropriate amount of lactose is added and mixed, a mixture of volatile oils of asarum, cassia twig and Schisandra chinensis is sprayed, and then mixed to obtain Xiaoqinglong granules.

[0013] According to the method for preparing Xiaoqinglong granules according to the second aspect, the method further comprises:

[0014] The second solution is concentrated, an appropriate amount of sucrose is added, mixed evenly to form granules, and dried, and a mixture of volatile oils of asarum, cassia twig and Schisandra chinensis is sprayed on and mixed evenly to form Xiaoqinglong granules.

[0015] According to the method for preparing Xiaoqinglong granules of the second aspect, the obtained medicinal residues are decocted with the white peony root, ephedra, and licorice root in water twice, the first time for 2 hours and the second time for 1.5 hours;

[0016] The pinellia tuber and dried ginger are crushed into coarse powder, and 70% ethanol is used as a solvent, and then percolation is performed after immersion.

[0017] According to the method for preparing Xiaoqinglong granules described in the second aspect, the time for distilling and extracting volatile oil is 6-10 hours, preferably 8 hours.

[0018] The method for preparing Xiaoqinglong granules according to the second aspect is characterized in that the volatile oil mixture at least includes the characteristic components of Schisandra chinensis var. aestivum, Schisandrin A and Schisandrin A.

[0019] The third aspect of the present invention further provides Xiaoqinglong granules, which are prepared according to the method for preparing Xiaoqinglong granules described in the second aspect.

[0020] The fourth aspect of the present invention also provides the use of Schisandra chinensis, Schisandra chinensis extract, the Xiaoqinglong decoction described in the first aspect or the Xiaoqinglong granules described in the third aspect in the preparation of drugs for treating respiratory diseases.

[0021] The fifth aspect of the present invention also provides the use of Schisandra chinensis, Schisandra chinensis extract, the Xiaoqinglong decoction described in the first aspect, or the Xiaoqinglong granules described in the third aspect in the preparation of drugs for inhibiting novel coronavirus infection.

[0022] The sixth aspect of the present invention also provides the use of Schisandra chinensis and Schisandra chinensis extract in a traditional Chinese medicine compound preparation for treating respiratory system diseases.

[0023] The applicant determined through qualitative and quantitative studies that there are significant differences in the chemical components of Schisandra chinensis (northern Schisandrae Chinensis) and Schisandra chinensis chinensis; a bronchial asthma model was established with mice, and it was determined that the pharmacological effect of Schisandra chinensis chinensis chinensis in treating bronchial asthma is better than that of Schisandra chinensis chinensis. Similarly, a bronchial asthma model was established with mice, and it was found that the pharmacological effects of Xiaoqinglong Decoction and Xiaoqinglong Granules after replacement were better than the original prescription. This combination optimization method is also applicable to the formulation optimization of the classic prescription containing Schisandra chinensis for treating bronchial asthma. That is, Schisandra chinensis chinensis chinensis, Schisandra chinensis chinensis extract, Xiaoqinglong Decoction prepared with Schisandra chinensis chinensis extract, or Xiaoqinglong Granules prepared with Schisandra chinensis chinensis extract all have good application effects in the preparation of drugs for the treatment of respiratory diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] These and / or other aspects and advantages of the present invention will become apparent and readily understood from the following description of the preferred embodiments in conjunction with the accompanying drawings, in which:

[0025] Figure 1 UPLC-Q-TOF-MS total ion chromatogram of the extracts of Schisandra chinensis (a) and Schisandra chinensis chinensis (b) provided in Example 1 of the present invention;

[0026] Figure 2 GC-MS total ion chromatograms of the volatile oils of Schisandra chinensis (a) and Schisandra chinensis chinensis (b) provided in Example 1 of the present invention;

[0027] Figure 3 A comparison chart of the levels of inflammatory factors in the serum of mice in the experimental groups provided in Example 2 of the present invention;

[0028] Figure 4 This is an H&E staining image of mouse lung tissue provided in Example 2 of the present invention;

[0029] Figure 5 A comparison chart of the contents of various factors in mouse serum provided in Example 3 of the present invention;

[0030] Figure 6 A comparison chart of the contents of various factors in mouse lung tissue provided in Example 3 of the present invention;

[0031] Figure 7 This is the H&E staining picture of mouse lung tissue provided in Example 3 of the present invention. DETAILED DESCRIPTION

[0032] The technical solution of the present invention is further specifically described below by examples and in conjunction with the accompanying drawings. In the specification, the same or similar reference numerals indicate the same or similar components. The following description of the embodiments of the present invention with reference to the accompanying drawings is intended to explain the overall inventive concept of the present invention and should not be construed as a limitation of the present invention.

[0033] Xiaoqinglong Decoction comes from Zhang Zhongjing's Treatise on Febrile Diseases in the Eastern Han Dynasty. It is composed of eight herbs: ephedra, cinnamon twig, peony root, asarum, dried ginger, schisandra, roasted liquorice, and pinellia. It has the effects of dispelling cold, warming the lungs and eliminating fluid. It is a classic prescription for treating "external cold and internal fluid". Modern pharmacological studies have found that Xiaoqinglong Decoction has anti-allergic, anti-inflammatory, antitussive and antiasthmatic effects. It is mostly used in clinical treatment of respiratory diseases such as chronic bronchitis, bronchial asthma, and chronic obstructive pulmonary disease.

[0034] After reviewing the composition of Xiaoqinglong Decoction in the herbal books of all dynasties (see Table 1), it was found that Schisandra chinensis was always included in the prescription.

[0035] Table 1 The prescriptions of Xiaoqinglong Decoction in ancient herbal medicine

[0036]

[0037]

[0038]

[0039] Schisandra chinensis was first recorded in Er Ya and was listed as a top grade in Shen Nong's Herbal Classic. Before the Ming Dynasty, Schisandra chinensis (i.e. northern Schisandra chinensis) was the main herb, and there was no clear distinction between the effects of southern Schisandra chinensis and Schisandra chinensis. Since 1963, Schisandra chinensis has been included in the Pharmacopoeia of the People's Republic of China. The 2000 edition of the Pharmacopoeia of the People's Republic of China first included Schisandra chinensis (Turcz.) Baill. and Schisandra sphenanthera Rehd. et Wils. from the Magnoliaceae plant as two separate varieties, respectively called "Schisandra chinensis" and "Southern Schisandra chinensis", but the description of the effects of both is the same as "astringent, invigorating Qi and promoting fluid, tonifying the kidney and calming the mind", which has continued to the current edition of the Pharmacopoeia of the People's Republic of China.

[0040] The applicant found that the existing Xiaoqinglong decoction or granules all use Schisandra chinensis, and there is no research and application of Schisandra chinensis chinensis in respiratory diseases.

[0041] The applicant found through qualitative and quantitative studies that there are significant differences in the chemical components of Schisandra chinensis (northern Schisandrae Chinensis) and Schisandra chinensis chinensis; a bronchial asthma model was established with mice, and it was found that the pharmacological effect of southern Schisandrae Chinensis in treating bronchial asthma is better than that of Schisandra chinensis. In order to enhance the pharmacological effect of the classic prescription Xiaoqinglong Decoction, the "Schisandra chinensis" used in the prescription was replaced with an equal dose of southern Schisandrae Chinensis, and all the medicinal materials in the prescription were inspected according to the content determination method under the corresponding medicinal materials in the current edition of the "Pharmacopoeia of the People's Republic of China", and all met the requirements of the pharmacopoeia and were qualified products. Similarly, a bronchial asthma model was established with mice, and it was found that the pharmacological effect of Xiaoqinglong Decoction after the replacement was better than the original prescription. This combination optimization method is also applicable to the optimization of the classic prescription containing Schisandrae Chinensis for the treatment of bronchial asthma, or other Chinese medicine compound prescriptions containing Schisandrae Chinensis.

[0042] Specifically, the present application discloses a Xiaoqinglong decoction, which is prepared with the following Chinese medicinal raw materials: ephedra, cinnamon twig, white peony root, dried ginger, asarum, roasted licorice, French pinellia, and also includes Schisandra chinensis, with a weight ratio of 2:2:2:2:1:2:3:2.

[0043] In a specific embodiment, the Xiaoqinglong decoction is prepared with the following minimum weight ratio of Chinese medicinal raw materials: 154g of ephedra, 154g of cinnamon twig, 154g of white peony root, 154g of dried ginger, 77g of asarum, 154g of roasted licorice, 231g of pinellia, and also includes 154g of Schisandra chinensis.

[0044] The present application also discloses a method for preparing Xiaoqinglong granules, wherein the Xiaoqinglong granules are prepared from the following Chinese medicinal raw materials: ephedra, cinnamon twig, white peony root, dried ginger, asarum, roasted licorice, pinellia tuber, and southern schisandra, with a weight ratio of 2:2:2:2:1:2:3:2; in a specific embodiment, the Xiaoqinglong granules are prepared from the following Chinese medicinal raw materials in the minimum weight ratio: ephedra 154g, cinnamon twig 154g, white peony root 154g, dried ginger 154g, asarum 77g, roasted licorice 154g, pinellia tuber 231g, and southern schisandra 154g;

[0045] The preparation method comprises:

[0046] (1) steam distilling Asarum, Cinnamomum cassiae and Schisandra chinensis together to extract a mixture of volatile oils from the three, and collecting the distilled aqueous solution in another device;

[0047] The obtained medicinal residues, the white peony root, ephedra, and licorice root are decocted in water for 2-3 times, each time for 2-4 hours, the decoctions are combined, filtered, the filtrate is combined with the distilled aqueous solution, and concentrated to obtain a first solution;

[0048] The pinellia tuber and dried ginger are crushed into coarse powder, and 60-90% ethanol is used as a solvent. After soaking, percolation is performed, and the percolation liquid is collected, and the ethanol is recovered and concentrated to an appropriate amount, and then combined with the first solution, allowed to stand, filtered, and the filtrate is concentrated to an appropriate amount (such as concentrated to 1000 mL) to obtain a second solution for standby use;

[0049] (2) The second solution is concentrated, spray-dried, an appropriate amount of lactose is added and mixed, a mixture of volatile oils of asarum, cassia twig and Schisandra chinensis is sprayed, and then mixed to obtain Xiaoqinglong granules.

[0050] In a specific embodiment, the method further comprises:

[0051] The second solution is concentrated, an appropriate amount of sucrose is added, mixed evenly to form granules, and dried, and a mixture of volatile oils extracted by distillation of asarum, cassia twig and Schisandra chinensis is sprayed on, mixed evenly to form Xiaoqinglong granules.

[0052] In another specific embodiment, when preparing Xiaoqinglong granules, the obtained medicinal residues and the white peony root, ephedra, and licorice are decocted in water twice, the first time for 2 hours and the second time for 1.5 hours;

[0053] The pinellia tuber and dried ginger are crushed into coarse powder, and 70% ethanol is used as a solvent, and then percolation is performed after immersion.

[0054] The time for extracting the volatile oil by distillation is 6-10 hours, preferably 8 hours.

[0055] The volatile oil mixture of Asarum, Cinnamon Twig and Schisandra chinensis contains at least the characteristic components of Schisandra chinensis, namely, anwuzhisu, schisandrin A and schisandrin A.

[0056] The present application also discloses a Xiaoqinglong granule, which is prepared by the aforementioned method for preparing Xiaoqinglong granules.

[0057] In order to determine the performance differences between Schisandra chinensis and Schisandra chinensis chinensis, the present application conducts relevant qualitative and quantitative analysis through specific examples.

[0058] Example 1: Chemical Difference Analysis of Schisandrae Chinensis and Schisandrae Chinensis

[0059] 1.1 Medicinal materials:

[0060] 15 batches of Schisandrae Chinensis were collected, including 3 batches from Heilongjiang, 5 batches from Liaoning, 4 batches from Jilin, 2 batches from Inner Mongolia, and 1 batch from Shandong; 15 batches of Schisandrae Chinensis South were collected, including 8 batches from Shaanxi, 3 batches from Shanxi, 2 batches from Henan, 1 batch from Anhui, and 1 batch from Yunnan. They were crushed separately, passed through a No. 3 sieve, sealed in plastic bags, and stored in electronic moisture-proof cabinets.

[0061] 1.2 Sample preparation:

[0062] Take about 0.25g of medicinal powder respectively, weigh accurately, put it in a 20mL stoppered conical bottle, add 10mL of anhydrous ethanol. Schisandra chinensis and Schisandrae Chinensis chinensis are ultrasonically treated (power 250W, frequency 30kHz) for 30min, taken out, cooled, added with extraction solvent to make up, shake well, filter, take the filtrate, pass through a 0.22μm microporous filter head, and use it for UPLC-Q-TOF / MS mass spectrometry analysis.

[0063] Then take 20g of Schisandra chinensis and Schisandra chinensis powder respectively in a 500mL round-bottom flask, add 300mL of distilled water and zeolite, and extract for 6h according to the steam distillation method of the four general rules in the 2020 edition of the "Pharmacopoeia of the People's Republic of China". Add ethyl acetate as the receiving solvent in the volatile oil extractor. Anhydrous sodium sulfate is used as the dehydrating agent for the extract. During the extraction process, the solution in the flask is always kept in a slightly boiling state. After the extraction is completed, cool it, collect the volatile oil, dilute it 4 times with ethyl acetate, and use it for GC-MS gas chromatography mass spectrometry analysis.

[0064] This application uses UPLC-Q-TOF / MS to identify the components of Schisandra chinensis and Schisandra chinensis samples. Since lignans have better ionization effect in positive ion mode and no response in negative ion mode, this application uses positive ion mode for detection. According to the retention time of the reference substance, the primary and secondary mass spectrometry data, combined with the reference literature and the precise molecular mass of the components in the self-built database of UNIFI software, they are identified. The total ion flow diagram of the extracts of Schisandra chinensis and Schisandra chinensis is shown in Figure 1 (a) Figure 1 (b), the specific composition information is shown in Table 2.

[0065] A total of 31 biphenylcyclooctenes were deduced and identified from Schisandra chinensis (Schisandra chinensis). All 15 batches of Schisandra chinensis ethanol extracts from different origins contained these 31 lignans.

[0066] A total of 40 biphenylcyclooctene-type lignans were inferred and identified from Schisandra chinensis, including 5 tetrahydrofuran-type lignans, 2 diarylbutane-type lignans, and 2 cycloatun-type triterpenes. The chemical components of 15 batches of Schisandra chinensis ethanol extracts from different origins all contained these 40 lignan compounds.

[0067] The analysis of this application found that in terms of types, the composition of lignans in Schisandra chinensis is more diverse, involving diarylbutanes, tetrahydrofurans, and biphenylcyclooctadiene lignans. In terms of content, Schisandra chinensis and Schisandra chinensis have 12 common components, among which the content of schisandrin A in Schisandra chinensis is significantly higher than that in Schisandra chinensis, while the content of schisandrin C, schisandrin A, schisandrin B, and schisandrin A in Schisandra chinensis is significantly higher than that in Schisandra chinensis, and ansphenanthene is a unique component of Schisandra chinensis.

[0068]

[0069]

[0070]

[0071] The GC-MS total ion chromatograms of the volatile oils of Schisandra chinensis and Schisandra chinensis are shown in Figure 2 (a) and Figure 2 (b), the analysis of volatile components of Schisandra chinensis is shown in Table 3, and the analysis of volatile components of Schisandra chinensis is shown in Table 4. As can be seen from the figure and table, there are great differences in the volatile components of Schisandra chinensis and Schisandra chinensis. 37 volatile components were identified from the volatile oil of Schisandra chinensis, including 8 monoterpenes, 20 sesquiterpenes, and 9 aliphatic components, including olefins, alkynes, ketones, fatty acids and their esters. Among them, ylangene has the highest content. 34 volatile components were identified from the volatile oil of Schisandra chinensis, including 3 monoterpenes, 19 sesquiterpenes, and 12 aliphatic components. The specific component information is shown in Tables 3 and 4. Among them, tricyclo[2.2.1.0(2,6)]heptane,1,7-dimethyl-7-(4-methyl-3-pentenyl)-,(-)- has the highest content. Among them, there are 9 common volatile components in Schisandra chinensis and Schisandra chinensis, namely α-pinene, β-pinene, ylang-ylang, Bicyclo[5.2.0]nonane,2-methylene-4,8,8-trimethyl-4-viny, α-amorphene, γ-ylang-ylang, trans-α-bergamotol, trans-nerolidol, and cubediol.

[0072]

[0073]

[0074]

[0075] Example 2: Pharmacological Experiment

[0076] 2.1 Experimental Materials

[0077] 2.1.1 Experimental animals

[0078] SPF BALB / c mice, female, 8-10 weeks old, weighing 20±2g, were purchased from Beijing Sibeifu Biotechnology Co., Ltd., and the experimental animal production license number is SCXK(Beijing)2019-0010. Before the formal experiment, the mice were adaptively raised in a suitable environment for 7 days; lighting conditions: 12h light and dark alternation; temperature 24±2℃; relative humidity 40-60%.

[0079] 2.1.2 Preparation of drug solution

[0080] Sensitization solution: ovalbumin (OVA) and aluminum hydroxide (Al(OH)3) were added to physiological saline to prepare a suspension containing 500 μg OVA and 20 mg Al(OH)3 per 1 mL.

[0081] Preparation of Xiaoqinglong Granules (prepared by the pharmacopoeia method): Xiaoqinglong Granules were prepared into high-dose 0.8 g / mL and low-dose 0.4 g / mL solutions.

[0082] Preparation of the medicinal liquid of Xiaoqinglong Decoction containing southern and northern Schisandra chinensis: decoct ephedra, cinnamon twig, white peony root, dried ginger, liquorice, and Schisandra chinensis (or southern Schisandra chinensis) in water twice (add 8 times the water for the first time and extract for 60 minutes; add 6 times the water for the second time and extract for 40 minutes), filter, combine the two filtrates, and concentrate to a high dose equivalent to 0.8 g / mL of the crude drug and a low dose equivalent to 0.4 g / mL of the crude drug.

[0083] Preparation of Schisandra chinensis (Schisandra chinensis) medicinal material extract: Weigh 30g of Schisandra chinensis and Schisandra chinensis respectively, decoct twice in water (add 10 times the water for the first time and extract for 60min; add 8 times the water for the second time and extract for 40min), filter, combine the two filtrates, and concentrate to a high dose equivalent to 0.6g / mL of the crude drug and a low dose equivalent to 0.3g / mL of the crude drug.

[0084] 2.2 Experimental methods

[0085] 2.2.1 Grouping

[0086] The mice were divided into 13 groups, 8 mice in each group, including blank (Control) group, model (OVA) group, dexamethasone positive drug (DXMS, 10 mg / kg) group, high-dose Xiaoqinglong decoction containing Schisandra chinensis (XN-H, 8 g / kg) group, low-dose Xiaoqinglong decoction containing Schisandra chinensis (XN-L, 4 g / kg) group, high-dose Xiaoqinglong decoction containing South Schisandra chinensis (XS-H, 8 g / kg) group, low-dose Xiaoqinglong decoction containing South Schisandra chinensis (XS-L, 4 g / kg) group, high-dose Schisandra chinensis (NG-H, 6 g / kg) group, low-dose Schisandra chinensis (NG-L, 3 g / kg) group, high-dose South Schisandra chinensis (SG-H, 6 g / kg) group, low-dose South Schisandra chinensis (SG-L, 3 g / kg) group, high-dose Xiaoqinglong granule group (XG-H, 8 g / kg) group, and low-dose Xiaoqinglong granule group (XG-L, 4 g / kg).

[0087] 2.2.2 Construction of bronchitis asthma model and drug administration

[0088] On the 1st, 8th and 15th days, mice in other groups except the blank group were intraperitoneally injected with 0.2 mL of sensitizing solution, and the blank group was intraperitoneally injected with an equal amount of saline. From 21st to 27th days, mice in other groups except the control group were challenged with 3% OVA solution by aerosol for 7 consecutive days (30min / 10mL / time), and the blank group was aerosolized with an equal amount of saline. 1 hour before each aerosolization, each dosing group was gavaged with 0.2 mL of the drug according to the dose, and the blank group and the model group were gavaged with an equal amount of distilled water. 24 hours after the last aerosolization, the mice were eyeballed and blood was collected, and then the mice were killed and lung tissues were collected.

[0089] 2.2.3 Enzyme-linked immunosorbent assay

[0090] ELISA kits were used to detect the levels of proinflammatory cytokines (IL-4, IL-5, IL-13, TNF-α) in mouse serum.

[0091] 2.2.4 Pathological observation of lung tissue

[0092] 2.2.5 Data processing

[0093] All data were analyzed using IBM SPSS Statistics 25.0 software and the results are presented as mean ± standard deviation. The comparison among the groups was performed by one-way analysis of variance (ANOVA), and the LSD method was used for testing when the variances were equal, and the Tamhane T2 method was used for testing when the variances were unequal.

[0094] 2.3 Experimental Results

[0095] 2.3.1 Levels of inflammatory factors in mouse serum

[0096] A large number of inflammatory cytokines are often produced during the onset of bronchial asthma, and inflammatory factors can further promote respiratory system damage. By measuring the concentrations of inflammatory factors IL-4, IL-5, IL-13, and TNF-α in the serum of each group of mice, the effect of each drug on the expression level of each inflammatory factor in the OVA-induced bronchial asthma model was observed. Figure 3 The comparison chart of inflammatory factors in the serum of mice in the experimental groups is as follows: A: IL-4 content in different groups; B: IL-5 content in different groups; C: IL-13 content in different groups; D: TNF-α content in different groups. Compared with the Control group, ###p<0.001; compared with the Model group, *p<0.05, **p<0.01, ***p<0.001.

[0097] Depend on Figure 3 The results showed that compared with the blank group (Control), the contents of the four inflammatory factors in the model group were significantly increased (p<0.001), indicating that the bronchial asthma model was successfully established. Compared with the model group (OVA), the levels of inflammatory factors in other drug-administered groups showed a downward trend, among which the contents of each factor in the dexamethasone positive drug (DXMS) group were significantly reduced (p<0.001), close to the blank group.

[0098] Compared with the two types of Xiaoqinglong Decoctions, the levels of various inflammatory factors in the Xiaoqinglong Decoction containing southern Schisandra chinensis were lower than those in the Xiaoqinglong Decoction containing Schisandra chinensis. Among them, the levels of IL-4, IL-5, IL-13, and TNF-α in the Xiaoqinglong Decoction containing southern Schisandra chinensis were significantly lower than those in the high-dose Xiaoqinglong Decoction containing Schisandra chinensis group (p<0.05). The results indicate that the effect of Xiaoqinglong Decoction containing southern Schisandra chinensis in treating bronchial asthma is better than that of Xiaoqinglong Decoction containing northern Schisandra chinensis.

[0099] Compared with the Schisandra chinensis group, the levels of various inflammatory factors in the Schisandra chinensis group were lower than those in the Schisandra chinensis group; among them, the levels of IL-5 and TNF-α in the Schisandra chinensis group were significantly lower than those in the Schisandra chinensis group (p<0.05); compared with the low-dose Schisandra chinensis group, the levels of IL-4 and IL-13 in the Schisandra chinensis group were significantly lower (p<0.05).

[0100] In general, compared with the Schisandra group, the levels of various inflammatory factors in the serum of mice in the Schisandra group were generally lower than those in the Schisandra group, and there were certain differences. Therefore, it is believed that the effect of Schisandra in treating bronchial asthma is better than that of Schisandra; at the same time, compared with the group containing Xiaoqinglong decoction containing Schisandra, the levels of various inflammatory factors in the group containing Xiaoqinglong decoction containing Schisandra were generally lower than those in the group containing Xiaoqinglong decoction containing Schisandra, and there were differences. Therefore, it is believed that Xiaoqinglong decoction containing Schisandra has a better therapeutic effect on bronchial asthma. The specific levels of inflammatory factors are shown in Table 5.

[0101] Table 5 Content of each factor in mouse serum / pg·mL -1 ( n=4)

[0102]

[0103] Note: Compared with the Control group, ### p<0.001; compared with the Model group, * p<0.05, ** p<0.01, *** p<0.001.

[0104] Figure 4 These are H&E staining images of mouse lung tissue, specifically magnified 200×; A. Control group; B. Model group; C. DXMS group; D. XN-H group; E. XN-L group; F. XS-H group; G. XS-L group; H. NG-H group; I. NG-L group; J. SG-H group; K. SG-L group.

[0105] Depend on Figure 4 The results showed that the alveolar septa of the blank group (Control) were normal, with no obvious inflammatory cell infiltration. The alveolar structure of the model group (OVA) was severely damaged, with alveolar fusion, large-area thickening or collapse of the alveolar wall, and a large number of inflammatory cells infiltrating and agglomerating. The alveolar structure of the positive drug group (DXMS) was relatively clear, with no obvious thickening of the alveolar septa, and a small amount of inflammatory cell infiltration.

[0106] Compared with the model group, all drug-treated groups had a reversal effect on OVA-induced bronchial asthma, among which the inflammatory infiltration of lung tissue in the group containing Schisandra chinensis Xiaoqinglong Decoction and the group containing Schisandra chinensis Nanqi group was lighter than that in the group containing Schisandra chinensis Nanqi group.

[0107] Compared with the group containing Schisandra chinensis Xiaoqinglong Decoction and the group containing Schisandra chinensis Xiaoqinglong Decoction, the group containing Schisandra chinensis Xiaoqinglong Decoction had less inflammatory cell infiltration and clearer alveolar structure. The lung condition of the high-dose group containing Schisandra chinensis Xiaoqinglong Decoction was close to that of the positive drug group, and there was no obvious thickening of the alveolar septum.

[0108] Compared with the Schisandra chinensis group, the Schisandra chinensis group had lighter inflammatory infiltration of lung tissue and damage of alveolar structure, indicating that Schisandra chinensis has better therapeutic effect on bronchial asthma than Schisandra chinensis; compared with the Schisandra chinensis Xiaoqinglong Decoction group, the Schisandra chinensis Xiaoqinglong Decoction group had more complete alveolar structure and no large-area thickening of alveolar septa, indicating that the Schisandra chinensis Xiaoqinglong Decoction group has better therapeutic effect on bronchial asthma.

[0109] Example 3: Pharmacological Experiment

[0110] 3.1 Experimental Materials

[0111] 3.1.1 Experimental animals

[0112] SPF BALB / c mice, female, 8-10 weeks old, weighing 20±2g, were purchased from Beijing Sibeifu Biotechnology Co., Ltd., and the experimental animal production license number is SCXK(Beijing)2019-0010. Before the formal experiment, the mice were adaptively raised in a suitable environment for 7 days; lighting conditions: 12h light and dark alternation; temperature 24±2℃; relative humidity 40-60%.

[0113] 3.1.2 Preparation of drug solution

[0114] Sensitization solution: OVA and Al(OH)3 were added to normal saline to prepare a suspension containing 500 μg OVA and 20 mg Al(OH)3 per 1 mL.

[0115] Preparation of optimized Xiaoqinglong granule liquid medicine: extract volatile oil from asarum, cinnamon twig, and Schisandra chinensis (or southern Schisandra chinensis), collect the volatile oil, and collect the distilled aqueous solution in another device. Decoction ephedra, dried ginger, white peony root, roasted licorice, and French pinellia together twice (add 10 times water for the first time and extract for 60 minutes; add 6 times water for the second time and extract for 40 minutes), filter, combine the two filtrates and the aqueous solution obtained after steam distillation, concentrate to the equivalent of 0.6g / mL of crude drug, and mix in volatile oil to obtain liquid medicine.

[0116] 3.2 Experimental methods

[0117] 3.2.1 Grouping

[0118] The mice were divided into 5 groups, with 8 mice in each group, namely blank group (Control), model group (Model), dexamethasone positive drug group (DXMS, 10 mg / kg), group containing Schisandra chinensis Xiaoqinglong Granules (XN, 6 g / kg), and group containing Schisandra chinensis Xiaoqinglong Granules (XS, 6 g / kg).

[0119] 3.2.2 Model construction and drug administration of bronchial asthma

[0120] On the 1st, 8th and 15th days, mice in other groups except the blank group were intraperitoneally injected with 0.2 mL of sensitizing solution, and the blank group was intraperitoneally injected with an equal amount of saline. From 21st to 27th days, mice in other groups except the control group were challenged with 3% OVA solution by aerosol for 7 consecutive days (30min / 10mL / time), and the blank group was aerosolized with an equal amount of saline. 1 hour before each aerosolization, each dosing group was gavaged with 0.2 mL of the drug according to the dose, and the blank group and the model group were gavaged with an equal amount of distilled water. 24 hours after the last aerosolization, the mice were eyeballed and blood was collected, and then the mice were killed and lung tissues were collected.

[0121] 3.2.3 Enzyme-linked immunosorbent assay

[0122] ELISA kits were used to detect the levels of proinflammatory cytokines (IL-4, IL-5, IL-13, TNF-α) in mouse serum.

[0123] 3.2.4 Pathological observation of lung tissue

[0124] 3.2.5 Data processing

[0125] All data were analyzed using IBM SPSS Statistics 25.0 software and the results are presented as mean ± standard deviation. The comparison among the groups was performed by one-way analysis of variance (ANOVA), and the LSD method was used for testing when the variances were equal, and the Tamhane T2 method was used for testing when the variances were unequal.

[0126] 3.3 Experimental Results

[0127] 3.3.1 Levels of sexual dysfunction factors in mouse serum

[0128] Substitute the OD value of the sample into the regression equation of the standard curve to calculate the content of each factor in the sample. The specific data are shown in Tables 6 and 7 below.

[0129] The results show that the content of each inflammatory factor in the serum of mice in the model group was significantly higher than that in the blank group (p<0.001), and the levels of inflammatory factors in other groups tended to decrease compared with the model group. Among them, the levels of IL-4, IL-13, and TNF-α in each administration group were significantly lower than those in the model group (p<0.001 or p<0.01). However, although the IL-5 level in the XN group showed a downward trend compared with the model group, there was no significant difference. The levels of IL-4, IL-5, and IL-13 in the serum of the XS group were lower than those in the XN group, and were significant (p<0.01 or p<0.05). Among them, the levels of inflammatory factors IL-5 and TNF-α in the serum of the XS group were lower than those in the DXMS group.

[0130] The results of the visualization are as follows Figure 5 As shown, specifically A: IL-4 content in different groups; B: IL-5 content in different groups; C: IL-13 content in different groups; D: TNF-α content in different groups. Compared with the Control group, ###p<0.001; compared with the Model group, ***p<0.001, **p<0.01, *p<0.05; compared with the XN group, △△p<0.01, △p<0.05.

[0131] In the lung tissue of mice, the levels of various inflammatory factors in the model group were significantly higher than those in the blank group (p<0.001). Compared with the model group, the levels of IL-4, IL-5, and IL-13 in each drug-treated group were significantly reduced (p<0.001). Among them, the TNF-α content in the XS group was lower than that in the XN group, and it was significant (p<0.05). The visualization results are shown in Figure 6 As shown, specifically A: IL-4 content in different groups; B: IL-5 content in different groups; C: IL-13 content in different groups; D: TNF-α content in different groups. Compared with the Control group, ###p<0.001; compared with the Model group, ***p<0.001, **p<0.01, *p<0.05; compared with the XN group, △△p<0.01, △p<0.05.

[0132] Table 6 Content of each factor in mouse serum / pg·mL -1 ( n=6)

[0133]

[0134] Note: Compared with the Control group, ###p<0.001; compared with the Model group, ***p<0.001, **p<0.01, *p<0.05; compared with the XN group, △△p<0.01, △p<0.05.

[0135] Table 7 Content of each factor in mouse lung tissue / pg·mL -1 ( n=6)

[0136]

[0137] Note: Compared with the Control group, ###p<0.001; compared with the Model group, ***p<0.001, **p<0.01, *p<0.05; compared with the XN group, △p<0.05.

[0138] In summary, the two types of Xiaoqinglong granules prepared by the optimized process have good therapeutic effects on OVA-induced bronchial asthma mice, and the therapeutic effect of Xiaoqinglong containing Schisandra chinensis chinensis on bronchial asthma mice is better than that of Xiaoqinglong containing Schisandra chinensis chinensis.

[0139] H&E staining images were obtained from Figure 7As shown, specifically magnified 100×; A. Control group; B. Model group; C. DXMS group; D. XN group; E. XS group. The alveolar septa in the Control group were normal, with no obvious inflammatory cell infiltration. The alveolar structure in the Model group was severely damaged, the alveoli were fused, the alveolar wall was thickened in a large area, and a large number of inflammatory cells were infiltrated around the bronchial wall. Compared with the Model group, each drug-treated group had a reversal effect on OVA-induced bronchial asthma. The alveolar structure in the DXMS group was clearer, and the alveolar septa were not obviously thickened, which was similar to the Control group. The lung tissue structures of the XN group and the XS group were significantly improved compared with the Model group. Among them, the lung tissue structure of the XS group was clearer, there was no large-area alveolar wall thickening, and the inflammatory infiltration was milder, close to the DXMS group. In summary, it is shown that the liquid containing Schisandra chinensis and Xiaoqinglong granules in Schisandra chinensis has a therapeutic effect on OVA-induced bronchial asthma mice, and the liquid containing Xiaoqinglong granules in Schisandra chinensis has a better therapeutic effect.

[0140] The applicant determined through qualitative and quantitative studies that there are significant differences in the chemical components of Schisandra chinensis (northern Schisandrae Chinensis) and Schisandra chinensis chinensis; a bronchial asthma model was established with mice, and it was determined that the pharmacological effect of Schisandra chinensis chinensis chinensis in treating bronchial asthma is better than that of Schisandra chinensis chinensis. Similarly, a bronchial asthma model was established with mice, and it was found that the pharmacological effects of Xiaoqinglong Decoction and Xiaoqinglong Granules after replacement were better than the original prescription. This combination optimization method is also applicable to the formulation optimization of the classic prescription containing Schisandra chinensis for treating bronchial asthma. That is, Schisandra chinensis chinensis chinensis, extract of Schisandra chinensis chinensis chinensis, Xiaoqinglong Decoction prepared with extract of Schisandra chinensis chinensis chinensis, or Xiaoqinglong Granules prepared with extract of Schisandra chinensis chinensis chinensis chinensis have a good application effect in the preparation of drugs for treating respiratory diseases.

[0141] Schisandra chinensis, Schisandra chinensis extract, Xiaoqinglong decoction prepared with Schisandra chinensis extract, or Xiaoqinglong granules prepared with Schisandra chinensis extract can be used in the preparation of drugs for inhibiting novel coronavirus infection.

[0142] Furthermore, the application of Schisandra chinensis and Schisandra chinensis extract in traditional Chinese medicine compound preparations for treating respiratory diseases, such as chronic bronchitis, bronchial asthma, chronic obstructive pulmonary disease and other respiratory diseases, if the original traditional Chinese medicine compound preparation includes Schisandra chinensis, the optimized compound preparation can be obtained by replacing Schisandra chinensis with Schisandra chinensis and Schisandra chinensis extract to optimize the preparation process, so as to achieve better therapeutic effects.

[0143] Although some embodiments of the present general inventive concept have been shown and described, it will be appreciated by those skilled in the art that changes may be made to these embodiments without departing from the principles and spirit of the present general inventive concept, the scope of which is defined by the claims and their equivalents.

Claims

1. A Xiaoqinglong decoction, which is prepared from the following Chinese medicinal raw materials: ephedra, cinnamon twig, white peony root, dried ginger, asarum, roasted liquorice, and pinellia tuber, characterized in that: It also includes Schisandra chinensis chinensis, with a weight ratio of 2:2:2:2:1:2:3:

2.

2. A method for preparing Xiaoqinglong granules, wherein the Xiaoqinglong granules are prepared from the following Chinese medicinal raw materials: ephedra, cinnamon twig, white peony root, dried ginger, asarum, roasted liquorice, pinellia tuber, and southern schisandra chinensis, with a weight ratio of 2:2:2:2:1:2:3:2; It is characterized in that The preparation method comprises: (1) steam distilling the three medicinal materials of Asarum, Cinnamomum cassia and Schisandra chinensis together to extract a mixture of volatile oils from the three materials, and collecting the aqueous solution after distillation in another device; The obtained medicinal residues, the white peony root, ephedra, and licorice root are decocted in water for 2-3 times, each time for 2-4 hours, the decoctions are combined, filtered, the filtrate is combined with the distilled aqueous solution, and concentrated to obtain a first solution; The pinellia tuber and dried ginger are crushed into coarse powder, 60-90% ethanol is used as a solvent, and then percolation is performed after immersion, and the percolation liquid is collected, and the ethanol is recovered and concentrated to a suitable amount, and then combined with the first solution, allowed to stand, filtered, and the filtrate is concentrated to obtain a second solution for standby use; (2) The second solution is concentrated, spray-dried, an appropriate amount of lactose is added and mixed, a mixture of volatile oils of asarum, cassia twig and Schisandra chinensis is sprayed, and then mixed to obtain Xiaoqinglong granules.

3. The method for preparing Xiaoqinglong granules according to claim 2, characterized in that: The method further comprises: The second solution is concentrated, an appropriate amount of sucrose is added, mixed evenly to form granules, and dried, and a mixture of volatile oils of asarum, cassia twig and Schisandra chinensis is sprayed on and mixed evenly to form Xiaoqinglong granules.

4. The method for preparing Xiaoqinglong granules according to claim 2, characterized in that: The obtained medicinal residues, the white peony root, ephedra, and licorice root are decocted in water for 2 times, the first time for 2 hours and the second time for 1.5 hours; The pinellia tuber and dried ginger are crushed into coarse powder, and 70% ethanol is used as a solvent, and then percolation is performed after immersion.

5. The method for preparing Xiaoqinglong granules according to claim 2, characterized in that: The time for extracting the volatile oil by distillation is 6-10 hours, preferably 8 hours.

6. The method for preparing Xiaoqinglong granules according to claim 2, characterized in that: The volatile oil mixture at least includes the characteristic components of Schisandra chinensis chinensis, Schisandrin A and Schisandrin A.

7. A Xiaoqinglong granule prepared according to the method for preparing Xiaoqinglong granules according to any one of claims 2 to 5.

8. Use of Schisandra chinensis, Schisandra chinensis extract, the Xiaoqinglong decoction described in claim 1, and the Xiaoqinglong granules described in claim 7 in the preparation of medicines for treating respiratory diseases.

9. Use of Schisandra chinensis, Schisandra chinensis extract, the Xiaoqinglong decoction described in claim 1, or the Xiaoqinglong granules described in claim 7 in the preparation of drugs for inhibiting novel coronavirus infection.

10. Application of Schisandra chinensis and Schisandra chinensis extracts in traditional Chinese medicine compound preparations for the treatment of respiratory diseases.

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