Application of vitex negundo prevention preparation in preparation of medicine for treating new crown sequelae and preparation method of vitex negundo prevention preparation

Abstract

The invention discloses application of a vitex negundo prevention preparation in preparation of a medicine for treating new crown sequelae and a preparation method of the vitex negundo prevention preparation, and belongs to the field of medicines. The negundo chastetree fruit prevention preparation is prepared from eleven raw medicinal materials including negundo chastetree fruit, divaricate saposhnikovia root, notopterygium root, radix angelicae pubescentis, radix bupleuri, radix peucedani, ligusticum wallichii, fructus aurantii, poria cocos, platycodon grandiflorum, liquorice and the like, and has the effects of inducing sweat, dispelling exogenous evils, dispelling wind and clearing damp. The negundo chastetree prevention preparation is an effective formula for treating new crown sequelae, can effectively treat viral myocarditis in the recovery period of new crown patients, and can reduce serum concentration of liver function markers ALT and AST and kidney function markers Cre and BUN, so that liver and kidney functions are effectively protected, inflammatory response of acute respiratory distress syndrome is effectively inhibited, diarrhea is effectively treated, and the curative effect is good. And the traditional Chinese medicine composition has an exact treatment effect on the new crown sequelae.

Description

Technical Field

[0001] The present invention relates to the application of Jingfang preparations and their preparation methods, specifically to the application of Jingfang preparations in the preparation of drugs for treating COVID-19 sequelae and their preparation methods, belonging to the medical field. Background Art

[0002] The most common clinical symptoms of patients infected with the novel coronavirus are fever, fatigue, and dry cough. A small number of patients are accompanied by upper respiratory and digestive tract symptoms such as nasal congestion, runny nose, and diarrhea. Severe cases usually develop dyspnea after 1 week, and in severe cases, it further progresses to acute respiratory distress syndrome, septic shock, refractory metabolic acidosis, and coagulation dysfunction. The novel coronavirus infection is a multi-system disease, and the affected areas of the disease in patients are not limited to the lungs, but also cause varying degrees of damage to the kidneys, liver, heart, brain, and nerves of patients. It is reported that most critically ill patients are accompanied by organ function damage, mostly including heart injury, acute kidney injury, liver dysfunction, and pneumothorax. So far, there is no specific effective drug for the novel coronavirus infection, and the main treatment methods are isolation treatment and symptomatic supportive treatment.

[0003] Although some discharged patients have negative nucleic acid tests, reduced or significantly absorbed pneumonia, and normal body temperature, they may still have symptoms such as fatigue, shortness of breath, decreased appetite, diarrhea, and anxiety. Some patients will also develop pulmonary fibrosis due to poor absorption of lung inflammation, which affects their quality of life, especially the prognosis of the elderly and those with chronic underlying diseases is poor. In addition, cured patients will also suffer from various organ damage sequelae caused by the novel coronavirus infection, such as heart injury, acute kidney injury, liver dysfunction, nervous system injury, and digestive system injury. According to a latest study released by the University Hospital of Frankfurt, the hearts of most people who have recovered from COVID-19 are damaged. Even if the COVID-19 test is negative, 78% of the participants still have heart problems, and the most common long-term damage factor is myocarditis. Therefore, the treatment of COVID-19 sequelae needs to attract more attention and emphasis.

[0004] Traditional Chinese medicine has played an important role in the diagnosis and treatment of this epidemic. Among them, the traditional Chinese medicine preparation Jingfang Keli has been recommended for the treatment of new coronavirus infections with the syndrome of damp-cold stagnation in the lungs in the prevention and control plans for new coronavirus infections in many provinces. The Jingfang preparation is derived from the ancient prescription Jingfang Baidu San, which is made from eleven raw medicinal materials including Schizonepeta tenuifolia Briq., Saposhnikovia divaricata (Turcz.) Schischk., Notopterygium incisum Ting ex H. T. Chang, Angelica pubescens Maxim. f. biserrata Shan et Yuan, Bupleurum chinense DC., Peucedanum praeruptorum Dunn, Ligusticum chuanxiong Hort., Aurantii Fructus Immaturus, Poria cocos (Schw.) Wolf, Platycodon grandiflorus (Jacq.) A. DC., and Glycyrrhiza uralensis Fisch. It has the effects of inducing sweating to relieve exterior syndrome, dispersing wind and removing dampness, and is clinically used to treat symptoms such as wind-cold cold, headache and body pain, aversion to cold without sweating, nasal congestion and runny nose, and cough with white phlegm. In the formula, the monarch drugs Schizonepeta tenuifolia Briq. and Saposhnikovia divaricata (Turcz.) Schischk. are pungent and warm in nature and can induce sweating to relieve exterior syndrome and disperse wind pathogens, removing the leading cause of various diseases; the minister drugs Notopterygium incisum Ting ex H. T. Chang and Angelica pubescens Maxim. f. biserrata Shan et Yuan are pungent and warm in nature and can disperse wind-cold-damp pathogens throughout the body. Ligusticum chuanxiong Hort. promotes blood circulation and dispels wind, and Bupleurum chinense DC. is pungent and disperses to relieve muscle soreness, assisting Notopterygium incisum Ting ex H. T. Chang and Angelica pubescens Maxim. f. biserrata Shan et Yuan to expel exogenous pathogens and relieve pain. Aurantii Fructus Immaturus lowers qi, Platycodon grandiflorus (Jacq.) A. DC. opens the lungs, Peucedanum praeruptorum Dunn reduces phlegm, and Poria cocos (Schw.) Wolf promotes diuresis, all of which are adjuvant drugs, and Glycyrrhiza uralensis Fisch. harmonizes all the drugs and serves as the guiding drug. When all the drugs are combined, sweating occurs and fever subsides, blood circulation is promoted and wind is eliminated and pain stops, the lungs function smoothly, phlegm and dampness are removed, cough stops, and all symptoms of the cold are eliminated and the disease is cured. In recent years, with the in-depth development of the research on traditional Chinese medicine preparations, more and more effects of Jingfang preparations have been discovered.

[0005] During the process of technological upgrading of the Jingfang preparation, the inventor found that the Jingfang preparation has an obvious effect on the treatment of sequelae of COVID-19. Pharmacodynamic experiments show that the Jingfang preparation is an effective formula for the treatment of sequelae of new coronavirus infection: it can effectively treat viral myocarditis, reduce the serum concentrations of liver function markers ALT and AST and renal function markers Cre and BUN, thereby effectively protecting liver and kidney functions, effectively inhibiting the inflammatory response of acute respiratory distress syndrome, and at the same time effectively treating diarrhea, treating both the symptoms and the root cause, and having a definite and significant therapeutic effect on the sequelae of COVID-19. Summary of the Invention

[0006] The purpose of the present invention is to provide an application of a Jingfang preparation in the preparation of a drug for treating sequelae of COVID-19. The prescription of the Jingfang preparation in the present invention is derived from the ancient prescription Jingfang Baidu San, and a series of Jingfang preparations such as Jingfang Keli and Jingfang tablets are prepared relying on existing technologies. The use described in the present invention was discovered during the clinical application of the company's drugs and was later verified by relevant pharmacodynamic experiments, having great commercial value.

[0007] The application of a Jingfang preparation in the preparation of a drug for treating sequelae of COVID-19, wherein the Jingfang preparation is made from the following traditional Chinese medicine components:

[0008]

[0009] Preferably:

[0010]

[0011] The test results show that Jingfang preparations can effectively treat viral myocarditis, reduce the serum concentrations of liver function markers ALT and AST and renal function markers Cre and BUN, thereby effectively protecting liver and kidney functions, effectively inhibiting the inflammatory response of acute respiratory distress syndrome, and at the same time effectively treating diarrhea, treating both the symptoms and the root causes, and having a definite and significant therapeutic effect on COVID-19 sequelae.

[0012] Another object of the present invention is to provide a traditional Chinese medicine oral preparation containing the above-mentioned traditional Chinese medicine composition and a preparation method thereof, and the traditional Chinese medicine oral preparation is one of granules, tablets, capsules and microcapsules.

[0013] The preparation method of the traditional Chinese medicine oral preparation includes the following steps:

[0014] A. The volatile oils of 7 herbs including Schizonepeta tenuifolia, Saposhnikovia divaricata, Notopterygium incisum, Angelica pubescens, Peucedanum praeruptorum, Ligusticum chuanxiong and Aurantii Fructus Immaturus are extracted respectively to obtain 7 kinds of volatile oils of herbs, and the residues after distillation, the aqueous solutions after distillation of Ligusticum chuanxiong and Aurantii Fructus Immaturus are reserved;

[0015] B. Take the volatile oils obtained in step A and respectively make inclusion compounds with β-cyclodextrin;

[0016] C. Prepare a 20-30% ethanol solution as a solvent with the aqueous solutions after distillation of Ligusticum chuanxiong and Aurantii Fructus Immaturus obtained in step A, and percolate the residues of Ligusticum chuanxiong and Aurantii Fructus Immaturus and Poria cocos obtained in step A to obtain a percolate for reserve;

[0017] D. Decoct the residues of Schizonepeta tenuifolia, Saposhnikovia divaricata, Notopterygium incisum, Angelica pubescens, Peucedanum praeruptorum obtained in step A and the remaining three herbs of Bupleurum chinense, Platycodon grandiflorum and Glycyrrhiza uralensis with water for 2-3 times, 1.5-2.5 hours each time, combine the decoctions, filter, and concentrate the filtrate to an extract with a relative density of 1.18-1.25 at 60-70 °C;

[0018] E. Combine the percolate obtained in step C and the extract obtained in step D, mix well, let stand, filter, and concentrate the filtrate to an extract with a relative density of 1.22-1.28 at 50-60 °C;

[0019] F. Take the extract obtained in step E and the β-cyclodextrin inclusion compound obtained in step B, and directly or add pharmaceutically acceptable excipients through conventional processes to prepare an oral pharmaceutical preparation.

[0020] The preferred dosage form of the present invention is granules, and the preparation method of the granules includes the following steps:

[0021] A. The volatile oils of 7 herbs including Schizonepeta tenuifolia, Saposhnikovia divaricata, Notopterygium incisum, Angelica pubescens, Peucedanum praeruptorum, Ligusticum chuanxiong and Aurantii Fructus Immaturus are extracted respectively to obtain 7 kinds of volatile oils of herbs, and the residues after distillation, the aqueous solutions after distillation of Ligusticum chuanxiong and Aurantii Fructus Immaturus are reserved;

[0022] B. Take the volatile oils obtained in step A and respectively make inclusion compounds with β-cyclodextrin;

[0023] C. Prepare a 25% ethanol solution with the aqueous solution after distillation of the Chuanxiong and Zhike obtained in step A as the solvent, and perform percolation on the medicinal residues of Chuanxiong and Zhike and Poria cocos obtained in step A to obtain a percolate for standby;

[0024] D. Decoct the medicinal residues of Schizonepeta tenuifolia, Saposhnikovia divaricata, Notopterygium incisum, Angelica pubescens, and Peucedanum praeruptorum obtained in step A and the remaining three flavors of Bupleurum chinense, Platycodon grandiflorum, and Glycyrrhiza uralensis three times with water for 2 hours each time. Combine the decoctions, filter, and concentrate the filtrate to an extract with a relative density of 1.23 at 60 - 70 °C;

[0025] E. Combine the percolate obtained in step C and the extract obtained in step D, mix well, let stand, filter, and concentrate the filtrate to an extract with a relative density of 1.26 at 50 - 60 °C;

[0026] F. Take the extract obtained in step E, perform belt vacuum drying under the conditions of a vacuum degree of -0.08 MPa - -0.10 Mpa and a drying temperature of 63 °C, pulverize it into fine powder, sieve it, add the β - cyclodextrin clathrate obtained in step B, mix well to obtain the fine powder of Jingfang extract, add excipients with a weight ratio of sucrose powder: hydroxypropyl starch: mannitol = 3:2:0.5 according to the formulated amount, mix well, make into granules, dry, and size the granules to obtain the product.

[0027] Another preferred dosage form of the present invention is a pellet preparation. The preparation method of the microcapsule preparation includes the following steps:

[0028] A. Extract volatile oils from seven medicinal materials of Schizonepeta tenuifolia, Saposhnikovia divaricata, Notopterygium incisum, Angelica pubescens, Peucedanum praeruptorum, Chuanxiong, and Zhike respectively to obtain seven kinds of volatile oils from the medicinal materials. Reserve the medicinal residues after distillation and the aqueous solution after distillation of Chuanxiong and Zhike;

[0029] B. Take the volatile oils obtained in step A and respectively prepare clathrates with β - cyclodextrin;

[0030] C. Prepare a 28% ethanol solution with the aqueous solution after distillation of the Chuanxiong and Zhike obtained in step A as the solvent, and perform percolation on the medicinal residues of Chuanxiong and Zhike and Poria cocos obtained in step A to obtain a percolate for standby;

[0031] D. Decoct the medicinal residues of Schizonepeta tenuifolia, Saposhnikovia divaricata, Notopterygium incisum, Angelica pubescens, and Peucedanum praeruptorum obtained in step A and the remaining three flavors of Bupleurum chinense, Platycodon grandiflorum, and Glycyrrhiza uralensis twice with water for 2.5 hours each time. Combine the decoctions, filter, and concentrate the filtrate to an extract with a relative density of 1.22 at 60 - 70 °C;

[0032] E. Combine the percolate obtained in step C and the extract obtained in step D, mix well, let stand, filter, and concentrate the filtrate to an extract with a relative density of 1.25 at 50 - 60 °C;

[0033] F. Take the extract obtained in step E, add the cyclodextrin clathrate obtained in step B, mix well, make into granules, perform belt vacuum drying, pulverize to obtain the fine powder of Jingfang extract for standby;

[0034] G. Weigh the fine powder of Schizonepeta tenuifolia and Ledebouriella seseloides extract, capsule material, anti-adhesive agent and plasticizer according to the formula amount. Add the capsule material, anti-adhesive agent and plasticizer to purified water, heat and stir at 54 °C until dissolved, and prepare a capsule material solution with a mass fraction of 33%. Cool to room temperature, add the fine powder of Schizonepeta tenuifolia and Ledebouriella seseloides extract and emulsifier under stirring, and homogenize and emulsify to obtain an emulsion for standby.

[0035] H. Spray-dry the emulsion in step G under the conditions of an inlet air temperature of 165 °C, a spray pressure of 0.40 MPa, and a feeding rate of 21.5 ml / min, collect the microcapsules, and cool to obtain the product.

[0036] Preferably, the conditions for belt vacuum drying in step F are a vacuum degree of -0.08 MPa to -0.10 MPa and a drying temperature of 56 °C.

[0037] Preferably, the capsule material in step G is sodium caseinate:maltodextrin = 3:2, the anti-adhesive agent is stearyl alcohol:titanium dioxide = 3:1, and the plasticizer is polyethylene glycol:citric acid = 3:1.

[0038] Preferably, the emulsifier in step C is a composite emulsifier of sucrose fatty acid ester:soybean phospholipid = 8:5 by weight, and the dosage is 1.18% of the total formulation amount by mass fraction.

[0039] To verify the effect of the Schizonepeta tenuifolia and Ledebouriella seseloides preparation of the present invention in treating the sequelae of COVID-19, the inventor carried out corresponding animal experimental studies. It should be noted that the samples selected for the animal experimental studies are those obtained from the representative formula and its preparation method of the present invention. Due to space limitations, the experiments and results of the products obtained from other formulas and preparation methods included in the present invention are not listed one by one here.

[0040] Experimental Example 1 Effect of Schizonepeta tenuifolia and Ledebouriella seseloides Preparation on Myocarditis in Mice

[0041] 1 Materials

[0042] 1.1 Experimental animals and feed SPF-grade male Balb / c mice, 6 - 8 weeks old, 60 mice, weighing 16 - 18 g, provided by Lunan Pharmaceutical Group Co., Ltd., experimental animal license number: SYXK(Lu)20180008. The mice were adaptively fed in a clean-grade animal laboratory for 1 week before the experiment, separated by gender, at a room temperature of 20 - 25 °C, a relative humidity of 40% - 60%, natural light, and free access to food and water.

[0043] 1.2 Instruments, reagents and drugs: AG285 electronic analytical balance (Mettler-Toledo), SE-LECTRA-E fully automatic biochemical analyzer (VWR Scientific), Thermo Scientific Medifuge small benchtop centrifuge (Thermo Fisher Scientific); Eagle medium (Vicente Biotechnology Co., Ltd.), mouse TNF-α and cTnI enzyme-linked immunosorbent assay kits (Nanjing Jiancheng Bioengineering Institute); the test drug was Jingfang Granules in Example 5 (produced by Lunan Hepu Pharmaceutical Co., Ltd.), and the positive control drug was Ribavirin Granules (batch number 20190701, Chenxin Pharmaceutical Co., Ltd.).

[0044] 2 Methods

[0045] 2.1 Grouping and model establishment: 60 healthy male Balb / c mice were randomly divided into 6 groups, with 10 mice in each group, namely the normal control group, the model control group, the Ribavirin positive control group (referred to as the "positive control group"), the high-dose Jingfang Granules group (referred to as the "test high-dose group"), the medium-dose Jingfang Granules group (referred to as the "test medium-dose group") and the low-dose Jingfang Granules group (referred to as the "test low-dose group"). CVB3 virus was cultured using Eagle medium. Except for the normal control group, the mice in the other groups were intraperitoneally injected with 0.2 ml of 1×10 4 / ml CVB3 dilution, and the normal control group was intraperitoneally injected with 0.2 ml of Eagle medium.

[0046] 2.2 Drug administration: 2 hours after virus inoculation, gavage administration was carried out in each group. According to the dose conversion coefficient of different animals in the appendix of the "Guiding Principles for Clinical Research of New Chinese Medicines", the positive control group was given Ribavirin 0.10 g / kg by gavage, and the test high-dose group, the test medium-dose group and the test low-dose group were given Jingfang Granules 9 g / kg, 4.5 g / kg and 2.25 g / kg by gavage respectively, which were equivalent to 2 times, 1 times and 0.5 times the human equivalent dose. The administration volume was 10 ml / kg. The normal control group and the model control group were given an equal volume of normal saline by gavage, once a day for 7 consecutive days.

[0047] 2.3 Detection indexes and methods: 7 days after drug administration, blood was taken from the mouse eyeballs, centrifuged at 4000 r / min for 10 min, and the serum was separated. The levels of cTnI (cardiac troponin I) and TNF-α (tumor necrosis factor α) in the serum were measured using an ELISA kit.

[0048] 2.4 Statistical analysis: SPSS19.0 statistical software was used for analysis, and the experimental data were expressed in the form of "mean ± standard deviation ". One-way ANOVA was used for comparison among multiple groups, and P < 0.05 was considered statistically significant.

[0049] 3 Results

[0050] Compared with the normal control group, the levels of cTnI and TNF-α in the serum of mice in the model control group were significantly increased, and the differences were statistically significant (P < 0.05), indicating that the animal model established in this experiment was successful. Compared with the model control group, the levels of cTnI and TNF-α in the serum of mice in the positive control group and the high, medium, and low-dose experimental groups were significantly decreased, and the differences were statistically significant (P < 0.05). The above results suggest that Jingfang Granules can effectively treat viral myocarditis. The results are shown in Table 1.

[0051] Table 1 Effects of Jingfang Granules on viral myocarditis in mice ( n = 10)

[0052]

[0053]

[0054] Note: Compared with the normal control group: P < 0.05 is indicated by " △ "; compared with the model control group: P < 0.05 is indicated by "*".

[0055] Experimental Example 2 Effects of Jingfang Preparation on Acute Liver Injury in Mice

[0056] 1 Materials

[0057] 1.1 Experimental animals and feed Healthy KM mice, half male and half female, 60 in number, weighing (20 ± 2) g, provided by Lunan Pharmaceutical Group Co., Ltd., experimental animal license number: SYXK (Lu) 20180008. Before the experiment, they were adaptively fed in a clean-grade animal laboratory for 1 week, separated by gender, at room temperature of 20 - 25°C, relative humidity of 40% - 60%, natural light, and free access to food and water.

[0058] 1.2 Instruments, reagents and drugs Thermo Scientific Medifuge small bench-top centrifuge (Thermo Fisher Scientific), AG285 electronic analytical balance (Mettler-Toledo); mouse ALT and AST enzyme-linked immunosorbent assay kits (Sigma), carbon tetrachloride (Shanghai Aladdin Biochemical Technology Co., Ltd.); the test drug was Jingfang Granules in Example 5 (produced by Lunan Hopeful Pharmaceutical Co., Ltd.), and the positive control drug was bifendate (batch number: 19058003, Shandong Health Pharmaceutical Co., Ltd.).

[0059] 2 Methods

[0060] 2.1 Grouping and model establishment: 60 healthy KM mice were randomly divided into 6 groups, namely the normal control group, the model control group, the bifendate positive control group (referred to as the "positive control group"), the high-dose Jingfang Granule group (referred to as the "experimental high-dose group"), the medium-dose Jingfang Granule group (referred to as the "experimental medium-dose group"), and the low-dose Jingfang Granule group (referred to as the "experimental low-dose group"), with 10 mice in each group, half male and half female. Except for the normal control group, the mice in the other groups were intraperitoneally injected with 0.2% carbon tetrachloride at a dose of 0.1 ml / 10 g. After 24 hours, an acute liver injury model was established. The normal control group was intraperitoneally injected with an equal dose of peanut oil. 0.5 mL of blood samples were collected (5 mice were randomly selected from each group and blood was taken from the orbital cavity), and the ALT (alanine aminotransferase) and AST (aspartate aminotransferase) in the serum were measured. The levels in each model group were significantly higher than those in the normal control group, indicating that the model was successfully established.

[0061] 2.2 Drug administration: On the 3rd day after the successful establishment of the model, gavage administration was carried out in each group. According to the dose conversion coefficient of different animals in the appendix of the "Guiding Principles for Clinical Research of New Chinese Medicines", the positive control group was given bifendate by gavage at a dose of 0.15 g / kg, and the experimental high-dose group, the experimental medium-dose group, and the experimental low-dose group were given Jingfang Granule by gavage at doses of 9 g / kg, 4.5 g / kg, and 2.25 g / kg respectively, which were equivalent to 2 times, 1 time, and 0.5 times the equivalent human dose. The administration volume was 10 ml / kg. The model control group was given an equal volume of normal saline by gavage once a day for 8 consecutive days.

[0062] 2.3 Detection indexes and methods: 1 hour after drug administration on the 8th day, the mice were bled from the orbital cavity, and 1.5 mL of blood samples were taken into centrifuge tubes, allowed to stand for 30 min, and centrifuged at 10000 r / min for 20 min to separate the serum. The ALT and AST levels were measured according to the instructions of the ELISA kit.

[0063] 2.4 Statistical analysis: SPSS 19.0 statistical software was used for analysis. The experimental data were expressed in the form of "mean ± standard deviation ". One-way ANOVA was used for comparison among multiple groups, and P < 0.05 was considered to indicate statistically significant differences.

[0064] 3 Results

[0065] Compared with the normal control group, the levels of ALT and AST in the serum of the mice in the model control group were significantly increased, and the differences were statistically significant (P < 0.05), indicating that the model establishment in this experiment was successful. Compared with the model control group, the levels of ALT and AST in the serum of the mice in the positive control group and the experimental high-, medium-, and low-dose groups were significantly decreased, and the differences were statistically significant (P < 0.05). The above results suggest that Jingfang Granule has a protective effect on CCl4-induced liver injury. The results are shown in Table 2.

[0066] Table 2 Effects of Jingfang Granules on Liver Injury in Mice( n = 10)

[0067]

[0068] Note: Compared with the normal control group: P < 0.05 is indicated by " △ "; compared with the model control group: P < 0.05 is indicated by "*".

[0069] Experimental Example 3 Effects of Drugs on Acute Kidney Injury in Mice

[0070] 1 Materials

[0071] 1.1 Experimental Animals and Feed Healthy KM mice, half male and half female, 60 in number, weighing (20 ± 2) g, provided by Lunan Pharmaceutical Group Co., Ltd., Experimental Animal License No.: SYXK(Lu)20180008. They were adaptively fed in a clean-grade animal laboratory for 1 week before the experiment, separated by gender, at room temperature of 20 - 25 °C, relative humidity of 40% - 60%, natural light, and free access to food and water.

[0072] 1.2 Instruments, Reagents and Drugs AG285 type electronic analytical balance (Mettler-Toledo), SE-LECTRA-E type automatic biochemical analyzer (Rittner Science), Thermo Scientific Medifuge small bench centrifuge (Thermo Fisher); cisplatin (Sigma), sodium pentobarbital (Jinan Hongbo Chemical Co., Ltd.), creatinine assay kit (Sigma), urea nitrogen assay kit (Quanzhou Ruixin Biotechnology Co., Ltd.); the test drug was Jingfang Granules in Example 5 (produced by Lunan Houpu Pharmaceutical Co., Ltd.), and the positive control drug was resveratrol (batch number: 19067002, Guangdong Bide Biotechnology Co., Ltd.).

[0073] 2 Methods

[0074] 2.1 Grouping and Modeling Take 60 healthy KM mice and randomly divide them into 6 groups, namely normal control group, model control group, resveratrol positive control group (abbreviated as "positive control group"), high-dose Jingfang Granules group (abbreviated as "test high-dose group"), medium-dose Jingfang Granules group (abbreviated as "test medium-dose group") and low-dose Jingfang Granules group (abbreviated as "test low-dose group"), with 10 mice in each group, half male and half female. Except for the normal control group, the mice in the other groups were intraperitoneally injected with 20 mg / kg cisplatin on the first day of the experiment to establish an acute kidney injury model in mice, and the normal control group was intraperitoneally injected with an equal volume of normal saline. Collect 0.5 mL of blood samples (randomly select 5 mice from each group and take blood from the orbital cavity), measure Cre (creatinine) and BUN (urea nitrogen) in the serum. The levels in each model group were significantly higher than those in the normal control group, indicating that the modeling was successful.

[0075] 2.2 Administration and Modeling On the second day after successful modeling, gavage administration was performed on each group. The doses were converted according to the dose conversion coefficients of different animals in the appendix of the "Guiding Principles for Clinical Research of New Chinese Medicines". The positive control group was gavaged with resveratrol at 0.20 g / kg, and the high-dose, medium-dose, and low-dose experimental groups were gavaged with Jingfang Granules at 9 g / kg, 4.5 g / kg, and 2.25 g / kg, respectively, which were equivalent to 2 times, 1 time, and 0.5 times the equivalent human dose. The administration volume was 10 ml / kg. The model control group was gavaged with an equal volume of normal saline once a day for 7 consecutive days.

[0076] 2.3 Detection Indexes and Methods Mice in each group were fasted for 12 hours before the last gavage administration. One hour after the last gavage, the mice were anesthetized by intraperitoneal injection of pentobarbital sodium (60 mg / kg). Blood was collected by eye socket enucleation. After centrifugation at 3000 r / min for 10 min, the serum was separated, and the contents of serum creatinine (Cre) and blood urea nitrogen (BUN) were measured using an automatic biochemical analyzer.

[0077] 2.4 Statistical Analysis SPSS 19.0 statistical software was used for analysis. The experimental data were expressed in the form of "mean ± standard deviation ". One-way analysis of variance was used for comparison among multiple groups, and P < 0.05 indicated that the difference was statistically significant.

[0078] 3 Results

[0079] Compared with the normal control group, the levels of Cre and BUN in the serum of mice in the model control group were significantly increased, and the differences were statistically significant (P < 0.05), indicating that the modeling of this experiment was successful. Compared with the model control group, the levels of Cre and BUN in the serum of mice in the positive control group and the high-, medium-, and low-dose experimental groups were significantly decreased, and the differences were statistically significant (P < 0.05). The above results suggest that Jingfang Granules have a protective effect on cisplatin-induced kidney injury. The results are shown in Table 3.

[0080] Table 3 Effects of Jingfang Granules on Kidney Injury in Mice ( n = 10)

[0081]

[0082]

[0083] Note: Compared with the normal control group: P < 0.05 is indicated by " △ "; compared with the model control group: P < 0.05 is indicated by "*".

[0084] Experimental Example 4 Effects of the Drug on Acute Respiratory Distress Syndrome (ARDS) in Rats

[0085] 1 Materials

[0086] 1.1 Experimental animals and feed: 60 healthy male SD rats, weighing (200±20) g, were provided by Lunan Pharmaceutical Group Co., Ltd. The experimental animal license number was SYXK (Lu) 20180008. One week before the experiment, they were adaptively fed in a clean-grade animal laboratory, separated by gender, at a room temperature of 20-25°C, a relative humidity of 40%-60%, natural light, and free access to food and water.

[0087] 1.2 Instruments, reagents and drugs: Thermo Scientific Medifuge small bench-top centrifuge (Thermo Fisher Scientific), -80°C low-temperature refrigerator (Qingdao Haier Special Electric Appliance Co., Ltd.), AG285 electronic analytical balance (Mettler-Toledo); ELISA kits for rat TNF-α and IL-1β (R&D), lipopolysaccharide LPS (Sigma), sodium pentobarbital (Jinan Hongbo Chemical Co., Ltd.); the test drug was Jingfang Granules in Example 5 (produced by Lunan Houpu Pharmaceutical Co., Ltd.), and the positive control drug was ambroxol hydrochloride (batch number: 19049001, Shanghai Boehringer Ingelheim Pharmaceuticals Co., Ltd.).

[0088] 2 Methods

[0089] 2.1 Grouping and modeling: 60 healthy male SD rats were randomly divided into 6 groups, with 10 rats in each group, namely the normal control group, the model control group, the ambroxol hydrochloride positive control group (referred to as the "positive control group"), the high-dose Jingfang Granules group (referred to as the "test high-dose group"), the medium-dose Jingfang Granules group (referred to as the "test medium-dose group"), and the low-dose Jingfang Granules group (referred to as the "test low-dose group"). Except for the normal control group, the rats in the other groups were instilled with 7.5 mg / kg of LPS (prepared with sterile normal saline) through the trachea, and then the rats were turned over to make the liquid evenly distributed in the two lungs of the rats to construct an ARDS model. The rats in the normal control group were instilled with an equal volume of sterile normal saline through the trachea. If the rats showed symptoms such as listlessness, less movement, loss of appetite, rapid breathing, and diarrhea, it was proved that the modeling was successful.

[0090] 2.2 Drug administration: 6 hours after modeling, according to the dose conversion coefficient of different animals in the appendix of the "Guiding Principles for Clinical Research of New Chinese Medicines", the rats in the positive control group were intraperitoneally injected with 0.04 g / kg of ambroxol hydrochloride injection, and the rats in the test high-dose group, test medium-dose group, and test low-dose group were respectively administered Jingfang Granules at 9 g / kg, 4.5 g / kg, and 2.25 g / kg, which was equivalent to 2 times, 1 time, and 0.5 times the equivalent human dose. The administration volume was 10 ml / kg. The normal control group and the model control group were given an equal volume of normal saline by gavage once a day for 21 consecutive days.

[0091] 2.3 Detection indicators and methods After the last administration to rats, sodium pentobarbital at a dose of 30 mg / kg was intraperitoneally injected. After anesthesia, tracheotomy was performed on the rats, a hollow catheter was inserted, the right bronchus was ligated, and the left lung was lavaged with 5 ml of cold saline through the left bronchus, repeated 3 times, with a recovery rate greater than 90%. The collected BALF (bronchoalveolar lavage fluid) was centrifuged at 3000 r / min for 10 min at 4°C, and the supernatant was stored in an ultra-low temperature refrigerator at -80°C. The contents of TNF-α and IL-1β were detected using TNF-α and IL-1β ELISA kits, and the operation was carried out strictly according to the kit instructions.

[0092] 2.4 Statistical analysis SPSS 19.0 statistical software was used for analysis. The experimental data were expressed in the form of "mean ± standard deviation ". One-way ANOVA was used for comparison among multiple groups, and P < 0.05 indicated that the difference was statistically significant.

[0093] 3 Results

[0094] Compared with the normal control group, the levels of TNF-α and IL-1β in the BALF of the model control group rats were significantly increased, and the differences were all statistically significant (P < 0.05), indicating that the model establishment in this experiment was successful. Compared with the model control group, the levels of TNF-α and IL-1β in the sera of the positive control group and the high, medium, and low-dose experimental groups rats were significantly decreased, and the differences were all statistically significant (P < 0.05). The above results suggest that Jingfang granules have a certain inhibitory effect on the inflammatory response of ARDS. The results are shown in Table 4.

[0095] Table 4 Effects of Jingfang granules on acute respiratory distress syndrome in rats ( n = 10)

[0096]

[0097] Note: Compared with the normal control group: P < 0.05 is indicated by " △ "; compared with the model control group: P < 0.05 is indicated by "*".

[0098] Experimental Example 5 Effects of the drug on diarrhea in mice

[0099] 1 Materials

[0100] 1.1 Experimental animals and feed SPF-grade KM mice, 60 in total, half male and half female, with a body weight of (20 ± 2) g, were provided by Lunan Pharmaceutical Group Co., Ltd. The experimental animal license number was: SYXK (Lu) 20180008. Before the experiment, the mice were adaptively fed in a clean-grade animal laboratory for 1 week, separated by gender, at a room temperature of 20 - 25°C, a relative humidity of 40% - 60%, natural light, and free access to food and water.

[0101] 1.2 Instruments, reagents and drugs: Automatic colony counter (Shanghai Haiheng Electromechanical Instrument Co., Ltd.), AG285 electronic analytical balance (Mettler-Toledo), Thermo Scientific Medifuge small bench centrifuge (Thermo Fisher Scientific), -80 °C low-temperature refrigerator (Qingdao Haier Special Electric Appliance Co., Ltd.), Bacterial incubator (Shanghai Jinghong Experimental Equipment Co., Ltd.), Anaerobic incubator (Shanghai Xinyi Instrument Co., Ltd.), Selective medium (Qingdao Haibo Biotech Co., Ltd.); Mouse diamine oxidase (DAO) ELISA kit (Xiamen Huijia Biotech Co., Ltd.), D-lactic acid detection and analysis kit (colorimetric method) (Wuhan Aimeijie Technology Co., Ltd.), SIgA kit (Institute of Isotopes, China Institute of Atomic Energy), Lincomycin (Shandong Ruiyang Pharmaceutical Co., Ltd.), PBS (Thermo Fisher Scientific); The test drug is Jingfang Granules in Example 5 (produced by Lunan Houpu Pharmaceutical Co., Ltd.), and the positive control drug is Bifid triple viable bacteria tablets (trade name Peifukang, batch number: 19075002, Shanghai Xinyi Pharmaceutical Factory Co., Ltd.).

[0102] 2 Methods

[0103] 2.1 Grouping and modeling: Take 60 SPF-grade KM mice. Among them, 50 mice were given intragastric administration of lincomycin hydrochloride at 0.4 mL / time, twice a day for 4 consecutive days. Observe that the mice showed listlessness, reduced activity, weight loss, slightly red perianal area, and diarrhea; at the same time, observe the defecation of the mice. The appearance of yellow-brown, loose and unformed feces, and the decrease in the number of Bifidobacterium and Lactobacillus in the fresh feces culture indicate that the modeling is successful. The 50 successfully modeled mice were randomly divided into a model control group, a Bifid triple viable bacteria tablets control group (referred to as the "positive control group"), a high-dose Jingfang Granules group (referred to as the "test high-dose group"), a medium-dose Jingfang Granules group (referred to as the "test medium-dose group") and a low-dose Jingfang Granules group (referred to as the "test low-dose group"), with 10 mice in each group, half male and half female. The remaining 10 mice were given intragastric administration of an equal volume of normal saline and set as the normal control group.

[0104] 2.2 Administration: Starting from the 5th day, intragastric administration was carried out in each group. According to the dose conversion coefficient of different animals in the appendix of the "Guiding Principles for Clinical Research of New Chinese Medicines", the positive control group was given Bifid triple viable bacteria tablets at 0.2 g / kg, and the test high-dose group, test medium-dose group and test low-dose group were given Jingfang Granules at 9 g / kg, 4.5 g / kg and 2.25 g / kg respectively, which is equivalent to 2 times, 1 time and 0.5 times the equivalent human dose. The administration volume was 10 ml / kg. The normal control group and the model control group were given intragastric administration of an equal volume of normal saline once a day for 14 consecutive days.

[0105] 2.3 Detection indexes and methods

[0106] 2.3.1 Intestinal flora detection After continuous administration for 14 days, the mice were sacrificed by cervical dislocation. 100 μg of cecal contents of each group of mice were collected aseptically, added with 800 μL of diluent and mixed well, and then diluted again. A suspension with a dilution of 1×10 -3 -1×10 -8 was inoculated into a selective medium and cultured in an incubator at 37°C and an anaerobic incubator for 24 h. Enterobacteria, enterococci, bifidobacteria, and lactobacilli were identified by colony morphology, biochemical reactions, and Gram staining. The identified bacteria were counted, and the logarithmic value was used to represent the number of dominant viable bacteria (lgCFU / g).

[0107] 2.3.2 Determination of diamine oxidase and D-lactic acid content After continuous administration for 14 days, the mice were sacrificed by cervical dislocation. 5 mL of blood was taken from the heart, centrifuged at 3000 r / min for 10 min, and the serum was taken to detect the contents of serum diamine oxidase (DAO) and D-lactic acid.

[0108] 2.3.3 Detection of intestinal mucosal SIgA (secretory immunoglobulin A) A 15-cm intestinal segment starting from 10 cm below the pylorus and extending towards the ileocecal region was taken, laid flat on filter paper, cut along the longitudinal section, and the surface substances of the intestinal mucosa were washed clean with sterile PBS buffer. Finally, a part of the mucus was gently scraped from the surface of the intestinal mucosa with a glass slide and placed in a 1.5-mL sterile EP tube for freezing and waiting for testing. According to the kit instructions, the biotin-avidin labeled enzyme-linked immunosorbent assay was used to analyze the SIgA level in the intestinal mucosa.

[0109] 2.4 Statistical analysis SPSS 19.0 statistical software was used for analysis, and the experimental data were expressed in the form of "mean ± standard deviation ". One-way ANOVA was used for comparison among multiple groups, and P < 0.05 was considered statistically significant.

[0110] 3 Results

[0111] 3.1 Comparison of intestinal flora Compared with the normal control group, the number of lactobacilli and bifidobacteria in the model control group decreased, while the number of enterobacteria and enterococci increased, and the differences were all statistically significant (P < 0.05). Compared with the model control group, the number of lactobacilli and bifidobacteria in the positive control group and the high, medium, and low-dose groups of the test increased, while the number of enterobacteria and enterococci decreased, and the differences were all statistically significant (P < 0.05). The results are shown in Table 5. The above results suggest that Jingfang Granule has a protective and reparative effect on the bacterial membrane barrier of the intestinal mucosa in mice with antibiotic-associated diarrhea.

[0112] 3.2 Determination of serum DAO and D-lactic acid levels Compared with the normal control group, the serum DAO and D-lactic acid levels in the model control group of mice were significantly increased, and the differences were statistically significant (P < 0.05). Compared with the model control group, the serum DAO and D-lactic acid levels in the positive control group and the high, medium, and low-dose experimental groups of mice were significantly decreased, and the differences were statistically significant (P < 0.05). The above results suggest that Jingfang Granule can effectively improve the intestinal barrier function of mice with antibiotic-associated diarrhea.

[0113] 3.3 Determination of intestinal mucus SIgA level Compared with the normal control group, the intestinal mucosal SIgA level in the model control group of mice was significantly decreased, and the difference was statistically significant (P < 0.05). Compared with the model control group, the intestinal mucosal SIgA levels in the positive control group and the high, medium, and low-dose experimental groups of mice were significantly increased, and the differences were statistically significant (P < 0.05). The above results suggest that Jingfang Granule has a positive promoting effect on improving the intestinal immune function of mice with antibiotic-associated diarrhea. The results are shown in Table 6.

[0114] In summary, Jingfang Granule can effectively treat antibiotic-associated diarrhea.

[0115] Table 5 Effects of Jingfang Granule on the intestinal flora quantity of mice ( n = 10)

[0116]

[0117] Note: Compared with the normal control group: P < 0.05 is indicated by " △ "; compared with the model control group: P < 0.05 is indicated by "*".

[0118] Table 6 Effects of Jingfang Granule on the serum levels of DAO, D-lactic acid and intestinal mucosal SIgA in mice ( n = 10)

[0119]

[0120] Note: Compared with the normal control group: P < 0.05 is indicated by " △ "; compared with the model control group: P < 0.05 is indicated by "*".

[0121] The above experimental results show that Jingfang Granule can effectively treat viral myocarditis, reduce the serum concentrations of liver function markers ALT and AST and renal function markers Cre and BUN, thus effectively protecting the liver and kidney functions, effectively inhibiting the inflammatory response of acute respiratory distress syndrome, and at the same time effectively treating diarrhea, treating both the symptoms and the root causes, and having a definite and significant therapeutic effect on the sequelae of COVID-19. Detailed implementation methods

[0122] The present invention will be further illustrated by specific embodiments below. However, those skilled in the art should understand that the embodiments do not limit the present invention in any way.

[0123] Example 1 Preparation of Jingfang Granules

[0124]

[0125] A. The volatile oils of seven medicinal materials, namely Schizonepeta tenuifolia Briq., Saposhnikovia divaricata (Turcz.) Schischk., Notopterygium incisum Ting ex H. T. Chang, Angelica pubescens Maxim. f. biserrata Shan et Yuan, Peucedanum praeruptorum Dunn, Ligusticum chuanxiong Hort. and Aurantii Fructus Immaturus, were extracted separately to obtain seven kinds of volatile oils of medicinal materials. The residues after distillation and the aqueous solutions after distillation of Ligusticum chuanxiong Hort. and Aurantii Fructus Immaturus were reserved for later use.

[0126] B. The volatile oils obtained in step A were separately made into inclusion compounds by adding β - cyclodextrin.

[0127] C. The aqueous solutions after distillation of Ligusticum chuanxiong Hort. and Aurantii Fructus Immaturus obtained in step A were made into a 27% ethanol solution as a solvent, and percolation was carried out on the residues of Ligusticum chuanxiong Hort., Aurantii Fructus Immaturus and Poria cocos Wolf obtained in step A to obtain a percolate for later use.

[0128] D. The residues of Schizonepeta tenuifolia Briq., Saposhnikovia divaricata (Turcz.) Schischk., Notopterygium incisum Ting ex H. T. Chang, Angelica pubescens Maxim. f. biserrata Shan et Yuan, and Peucedanum praeruptorum Dunn obtained in step A and the other three medicinal materials, namely Bupleurum chinense DC., Platycodon grandiflorum (Jacq.) A. DC. and Glycyrrhiza uralensis Fisch., were decocted with water three times, each time for 2 hours. The decoctions were combined, filtered, and the filtrate was concentrated to an extract with a relative density of 1.24 at 60 - 70 °C.

[0129] E. The percolate obtained in step C and the extract obtained in step D were combined, mixed evenly, allowed to stand, filtered, and the filtrate was concentrated to an extract with a relative density of 1.24 at 50 - 60 °C.

[0130] F. The extract obtained in step E was taken, and under the conditions of a vacuum degree of - 0.08 MPa - - 0.10 Mpa and a drying temperature of 63 °C, it was dried by belt vacuum drying, crushed into fine powder, sieved, and the β - cyclodextrin inclusion compound obtained in step B was added and mixed evenly to obtain the fine powder of Jingfang extract. The formulated amount of sucrose powder, hydroxypropyl starch, and mannitol (weight ratio 4:2:1) was added, mixed evenly, made into granules, dried, and sized to make 10 kg, thus obtaining the product.

[0131] Example 2 Preparation of Jingfang Tablets

[0132]

[0133] A. The volatile oils of seven medicinal materials, namely Schizonepeta tenuifolia Briq., Saposhnikovia divaricata (Turcz.) Schischk., Notopterygium incisum Ting ex H. T. Chang, Angelica pubescens Maxim. f. biserrata Shan et Yuan, Peucedanum praeruptorum Dunn, Ligusticum chuanxiong Hort. and Aurantii Fructus Immaturus, were extracted separately to obtain seven kinds of volatile oils of medicinal materials. The residues after distillation and the aqueous solutions after distillation of Ligusticum chuanxiong Hort. and Aurantii Fructus Immaturus were reserved for later use.

[0134] B. The volatile oils obtained in step A were separately made into inclusion compounds by adding β - cyclodextrin.

[0135] C. Prepare a 24% ethanol solution with the aqueous solution after distillation of the Chuanxiong and Zhike obtained in step A as the solvent, and perform percolation on the medicinal residues of Chuanxiong and Zhike and Poria cocos obtained in step A to obtain a percolate for standby;

[0136] D. Decoct the medicinal residues of Schizonepeta tenuifolia, Saposhnikovia divaricata, Notopterygium incisum, Angelica pubescens, and Peucedanum praeruptorum obtained in step A and the other three herbs of Bupleurum chinense, Platycodon grandiflorum, and Glycyrrhiza uralensis twice with water, each time for 1.5 hours. Combine the decoctions, filter, and concentrate the filtrate to an extract with a relative density of 1.25 at 60 - 70 °C;

[0137] E. Combine the percolate obtained in step C and the extract obtained in step D, mix well, let stand, filter, and concentrate the filtrate to an extract with a relative density of 1.28 at 50 - 60 °C;

[0138] F. Take the extract obtained in step E, perform belt vacuum drying under the conditions of a vacuum degree of -0.08 MPa - -0.10 Mpa and a drying temperature of 63 °C, crush it into fine powder, sieve it, add the β - cyclodextrin clathrate obtained in step B, mix well to obtain the fine powder of Jingfang extract. Add the formulated amount of microcrystalline cellulose, low - substituted hydroxypropyl cellulose, and mannitol (weight ratio 4:3:1), mix well to make coarse granules, dry, crush, sieve, make granules, dry at low temperature, size, add 0.2% magnesium stearate and 0.1% talc powder, mix well, and press into 10,000 tablets to obtain.

[0139] Example 3 Preparation of Jingfang Microcapsules

[0140]

[0141] A. Extract the volatile oils from 7 herbs of Schizonepeta tenuifolia, Saposhnikovia divaricata, Notopterygium incisum, Angelica pubescens, Peucedanum praeruptorum, Chuanxiong, and Zhike respectively to obtain 7 kinds of volatile oils from the herbs. Reserve the medicinal residues after distillation and the aqueous solution after distillation of Chuanxiong and Zhike;

[0142] B. Take the volatile oils obtained in step A and respectively make clathrates with β - cyclodextrin;

[0143] C. Prepare a 29% ethanol solution with the aqueous solution after distillation of the Chuanxiong and Zhike obtained in step A as the solvent, and perform percolation on the medicinal residues of Chuanxiong and Zhike and Poria cocos obtained in step A to obtain a percolate for standby;

[0144] D. Decoct the medicinal residues of Schizonepeta tenuifolia, Saposhnikovia divaricata, Notopterygium incisum, Angelica pubescens, and Peucedanum praeruptorum obtained in step A and the other three herbs of Bupleurum chinense, Platycodon grandiflorum, and Glycyrrhiza uralensis twice with water, each time for 2.5 hours. Combine the decoctions, filter, and concentrate the filtrate to an extract with a relative density of 1.23 at 60 - 70 °C;

[0145] E. Combine the percolate obtained in step C and the extract obtained in step D, mix well, let stand, filter, and concentrate the filtrate to an extract with a relative density of 1.27 at 50 - 60 °C;

[0146] F. Take the extract obtained in step E, add the cyclodextrin clathrate obtained in step B, mix well, make into granules, and carry out belt vacuum drying under the conditions of a vacuum degree of -0.08 MPa to -0.10 MPa and a drying temperature of 55 °C, then pulverize to obtain the fine powder of Schizonepeta tenuifolia and Ledebouriella seseloides extract for standby;

[0147] G. Weigh the fine powder of Schizonepeta tenuifolia and Ledebouriella seseloides extract obtained in step F according to the formula amount, use the mixture of sodium caseinate:maltodextrin = 5:3 as the capsule material, octadecanol:titanium dioxide = 2:1 as the anti-sticking agent, and polyethylene glycol:citric acid = 4:1 as the plasticizer. Add the above capsule material, anti-sticking agent and plasticizer to purified water, heat and stir at 52 °C until dissolved, and prepare a capsule material solution with a mass fraction of 33.5%. Cool to room temperature, add the fine powder of Schizonepeta tenuifolia and Ledebouriella seseloides extract under stirring, add a compound emulsifier of 1.17% of the total formulation amount of sucrose fatty acid ester:soybean phospholipid = 8:5, and carry out homogenization emulsification to obtain an emulsion for standby;

[0148] H. Spray dry the emulsion in step G under the conditions of an inlet air temperature of 166 °C, a spray pressure of 0.40 MPa, and a feed rate of 22.5 ml / min, collect the microcapsules, and cool to obtain the product.

[0149] Preparation of Schizonepeta tenuifolia and Ledebouriella seseloides Capsules in Example 4

[0150]

[0151] A. Extract the volatile oils from 7 kinds of medicinal materials including Schizonepeta tenuifolia, Ledebouriella seseloides, Notopterygium incisum, Angelica pubescens, Peucedanum praeruptorum, Ligusticum chuanxiong and Aurantii Fructus Immaturus respectively to obtain 7 kinds of volatile oils from medicinal materials, and reserve the residues after distillation and the aqueous solutions after distillation of Ligusticum chuanxiong and Aurantii Fructus Immaturus;

[0152] B. Take the volatile oils obtained in step A and respectively add β-cyclodextrin to make clathrates;

[0153] C. Prepare a 29% ethanol solution as the solvent with the aqueous solutions after distillation of Ligusticum chuanxiong and Aurantii Fructus Immaturus obtained in step A, and carry out percolation on the residues of Ligusticum chuanxiong and Aurantii Fructus Immaturus and Poria cocos obtained in step A to obtain a percolate for standby;

[0154] D. Decoct the residues of Schizonepeta tenuifolia, Ledebouriella seseloides, Notopterygium incisum, Angelica pubescens and Peucedanum praeruptorum obtained in step A and the other three kinds of medicinal materials including Bupleurum chinense, Platycodon grandiflorum and Glycyrrhiza uralensis with water for 3 times, each time for 1.5 hours, combine the decoction liquids, filter, and concentrate the filtrate to an extract with a relative density of 1.20 at 60 - 70 °C;

[0155] E. Combine the percolate obtained in step C and the extract obtained in step D, mix well, let stand, filter, and concentrate the filtrate to an extract with a relative density of 1.24 at 50 - 60 °C;

[0156] F. Take the extract obtained in step E, and perform belt vacuum drying under the conditions of a vacuum degree of -0.08 MPa to -0.10 MPa and a drying temperature of 63 °C. Crush it into fine powder, sieve it, add the β-cyclodextrin clathrate obtained in step B, mix well, and obtain the fine powder of Schizonepeta tenuifolia and Ledebouriella seseloides extract. Add the formulated amount of starch and microcrystalline cellulose (weight ratio 6:1), mix well, granulate, dry, size, fill, polish in a polishing machine, and remove the damaged capsules to obtain the product.

[0157] Preparation of Schizonepeta tenuifolia and Ledebouriella seseloides Granules in Example 5

[0158]

[0159] A. Extract the volatile oils from 7 kinds of medicinal materials including Schizonepeta tenuifolia, Ledebouriella seseloides, Notopterygium incisum, Angelica pubescens, Peucedanum praeruptorum, Ligusticum chuanxiong and Aurantii Fructus Immaturus respectively to obtain 7 kinds of volatile oils from medicinal materials. The residues after distillation and the aqueous solutions after distillation of Ligusticum chuanxiong and Aurantii Fructus Immaturus are reserved for use.

[0160] B. Take the volatile oils obtained in step A and prepare clathrates with β-cyclodextrin respectively.

[0161] C. Prepare a 25% ethanol solution as a solvent from the aqueous solutions after distillation of Ligusticum chuanxiong and Aurantii Fructus Immaturus obtained in step A, and perform percolation on the residues of Ligusticum chuanxiong and Aurantii Fructus Immaturus and Poria cocos obtained in step A to obtain a percolate for reserve.

[0162] D. Decoct the residues of Schizonepeta tenuifolia, Ledebouriella seseloides, Notopterygium incisum, Angelica pubescens and Peucedanum praeruptorum obtained in step A and the other three kinds of medicinal materials including Bupleurum chinense, Platycodon grandiflorum and Glycyrrhiza uralensis with water for 3 times, 2 hours each time. Combine the decoction liquids, filter, and concentrate the filtrate to an extract with a relative density of 1.23 at 60 - 70 °C.

[0163] E. Combine the percolate obtained in step C and the extract obtained in step D, mix well, let stand, filter, and concentrate the filtrate to an extract with a relative density of 1.26 at 50 - 60 °C.

[0164] F. Take the extract obtained in step E, and perform belt vacuum drying under the conditions of a vacuum degree of -0.08 MPa to -0.10 MPa and a drying temperature of 63 °C. Crush it into fine powder, sieve it, add the β-cyclodextrin clathrate obtained in step B, mix well, and obtain the fine powder of Schizonepeta tenuifolia and Ledebouriella seseloides extract. Add the excipients with a formulated weight ratio of sucrose powder: hydroxypropyl starch: mannitol = 3:1:0.5, mix well, make into granules, dry, size, and obtain the product.

[0165] Preparation of Schizonepeta tenuifolia and Ledebouriella seseloides Capsules in Example 6

[0166]

[0167] A. Extract the volatile oils from 7 kinds of medicinal materials including Schizonepeta tenuifolia, Ledebouriella seseloides, Notopterygium incisum, Angelica pubescens, Peucedanum praeruptorum, Ligusticum chuanxiong and Aurantii Fructus Immaturus respectively to obtain 7 kinds of volatile oils from medicinal materials. The residues after distillation and the aqueous solutions after distillation of Ligusticum chuanxiong and Aurantii Fructus Immaturus are reserved for use.

[0168] B. Take the volatile oils obtained in step A and prepare inclusion complexes with β-cyclodextrin respectively;

[0169] C. Prepare a 26% ethanol solution with the aqueous solution after distillation of the Chuanxiong and Zhike obtained in step A as the solvent, and percolate the medicinal residues of Chuanxiong and Zhike and Poria cocos obtained in step A to obtain a percolate for standby;

[0170] D. Decoct the medicinal residues of Schizonepeta tenuifolia, Saposhnikovia divaricata, Notopterygium incisum, Angelica pubescens and Peucedanum praeruptorum obtained in step A and the remaining three herbs of Bupleurum chinense, Platycodon grandiflorum and Glycyrrhiza uralensis twice with water for 2 hours each time. Combine the decoction liquids, filter, and concentrate the filtrate to an extract with a relative density of 1.19 at 60 - 70 °C;

[0171] E. Combine the percolate obtained in step C and the extract obtained in step D, mix well, let stand, filter, and concentrate the filtrate to an extract with a relative density of 1.23 at 50 - 60 °C;

[0172] F. Take the extract obtained in step E, perform belt vacuum drying under the conditions of a vacuum degree of -0.08 MPa - -0.10 Mpa and a drying temperature of 63 °C, pulverize it into fine powder, sieve it, add the β-cyclodextrin inclusion complex obtained in step B, mix well to obtain the fine powder of Jingfang extract, add the formulated amount of starch, microcrystalline silica, and low-substituted hydroxypropyl cellulose (weight ratio 5:2:2), mix well, granulate, dry, size the granules, fill them into capsules, polish them in a polishing machine, and remove the damaged capsules to obtain the product.

[0173] Preparation of Jingfang Capsules in Example 7

[0174]

[0175] A. Extract volatile oils from 7 herbs of Schizonepeta tenuifolia, Saposhnikovia divaricata, Notopterygium incisum, Angelica pubescens, Peucedanum praeruptorum, Chuanxiong and Zhike respectively to obtain 7 kinds of volatile oils from the herbs. Reserve the medicinal residues after distillation and the aqueous solution after distillation of Chuanxiong and Zhike;

[0176] B. Take the volatile oils obtained in step A and prepare inclusion complexes with β-cyclodextrin respectively;

[0177] C. Prepare a 30% ethanol solution with the aqueous solution after distillation of the Chuanxiong and Zhike obtained in step A as the solvent, and percolate the medicinal residues of Chuanxiong and Zhike and Poria cocos obtained in step A to obtain a percolate for standby;

[0178] D. Decoct the medicinal residues of Schizonepeta tenuifolia, Saposhnikovia divaricata, Notopterygium incisum, Angelica pubescens and Peucedanum praeruptorum obtained in step A and the remaining three herbs of Bupleurum chinense, Platycodon grandiflorum and Glycyrrhiza uralensis twice with water for 1.5 hours each time. Combine the decoction liquids, filter, and concentrate the filtrate to an extract with a relative density of 1.19 at 60 - 70 °C;

[0179] E. Combine the percolate obtained in step C and the extract obtained in step D, mix well, let stand, filter, and concentrate the filtrate to an extract with a relative density of 1.25 at 50 - 60 °C;

[0180] F. Take the extract obtained in step E, and perform belt vacuum drying under the conditions of a vacuum degree of -0.08 MPa to -0.10 MPa and a drying temperature of 63°C. Crush it into fine powder, sieve it, add the β-cyclodextrin clathrate obtained in step B, mix well, and you will get the fine powder of Schizonepeta tenuifolia and Ledebouriella seseloides extract. Add the formulated amount of starch and microcrystalline silica (weight ratio 4:1), mix well, granulate, dry, size the granules, fill them into capsules, polish them in a polishing machine, and remove the damaged capsules to obtain the product.

[0181] Preparation of Schizonepeta tenuifolia and Ledebouriella seseloides Tablets in Example 8

[0182]

[0183] A. Extract the volatile oils from the seven medicinal materials of Schizonepeta tenuifolia, Ledebouriella seseloides, Notopterygium incisum, Angelica pubescens, Peucedanum praeruptorum, Ligusticum chuanxiong, and Aurantii Fructus Immaturus respectively to obtain seven kinds of volatile oils from the medicinal materials. The residues after distillation and the aqueous solutions after distillation of Ligusticum chuanxiong and Aurantii Fructus Immaturus are reserved for later use.

[0184] B. Take the volatile oils obtained in step A and prepare clathrates with β-cyclodextrin respectively.

[0185] C. Prepare a 22% ethanol solution with the aqueous solution after distillation of Ligusticum chuanxiong and Aurantii Fructus Immaturus obtained in step A as the solvent, and perform percolation on the residues of Ligusticum chuanxiong and Aurantii Fructus Immaturus and Poria cocos obtained in step A to obtain a percolate for later use.

[0186] D. Decoct the residues of Schizonepeta tenuifolia, Ledebouriella seseloides, Notopterygium incisum, Angelica pubescens, and Peucedanum praeruptorum obtained in step A and the other three medicinal materials of Bupleurum chinense, Platycodon grandiflorum, and Glycyrrhiza uralensis with water twice, each time for 2.5 hours. Combine the decoction liquids, filter, and concentrate the filtrate to an extract with a relative density of 1.24 at 60 - 70°C.

[0187] E. Combine the percolate obtained in step C and the extract obtained in step D, mix well, let it stand, filter, and concentrate the filtrate to an extract with a relative density of 1.22 at 50 - 60°C.

[0188] F. Take the extract obtained in step E, and perform belt vacuum drying under the conditions of a vacuum degree of -0.08 MPa to -0.10 MPa and a drying temperature of 63°C. Crush it into fine powder, sieve it, add the β-cyclodextrin clathrate obtained in step B, mix well, and you will get the fine powder of Schizonepeta tenuifolia and Ledebouriella seseloides extract. Add the formulated amount of starch, dextrin, and sucrose (weight ratio 3:2:1), mix well, make into coarse granules, dry, crush, sieve, make granules, dry at low temperature, size the granules, add 0.3% magnesium stearate, mix well, and press into 10,000 tablets to obtain the product.

[0189] Preparation of Schizonepeta tenuifolia and Ledebouriella seseloides Granules in Example 9

[0190]

[0191] A. The volatile oils of 7 medicinal materials, namely Schizonepeta tenuifolia Briq., Saposhnikovia divaricata (Turcz.) Schischk., Notopterygium incisum Ting ex H. T. Chang, Angelica pubescens Maxim. f. biserrata Shan et Yuan, Peucedanum praeruptorum Dunn, Ligusticum chuanxiong Hort. and Aurantii Fructus Immaturus, are extracted respectively to obtain 7 kinds of volatile oils of medicinal materials. The residues after distillation and the aqueous solutions after distillation of Ligusticum chuanxiong Hort. and Aurantii Fructus Immaturus are reserved for later use.

[0192] B. Take the volatile oils obtained in step A and prepare inclusion complexes with β-cyclodextrin respectively.

[0193] C. Prepare a 20% ethanol solution with the aqueous solutions after distillation of Ligusticum chuanxiong Hort. and Aurantii Fructus Immaturus obtained in step A as the solvent, and percolate the residues of Ligusticum chuanxiong Hort., Aurantii Fructus Immaturus and Poria cocos Wolf obtained in step A to obtain a percolate for later use.

[0194] D. Decoct the residues of Schizonepeta tenuifolia Briq., Saposhnikovia divaricata (Turcz.) Schischk., Notopterygium incisum Ting ex H. T. Chang, Angelica pubescens Maxim. f. biserrata Shan et Yuan and Peucedanum praeruptorum Dunn obtained in step A together with the remaining three medicinal materials, namely Bupleuri Radix, Platycodonis Radix and Glycyrrhizae Radix, 3 times with water for 1.5 hours each time. Combine the decoctions, filter, and concentrate the filtrate to an extract with a relative density of 1.18 at 60 - 70 °C.

[0195] E. Combine the percolate obtained in step C and the extract obtained in step D, mix well, let stand, filter, and concentrate the filtrate to an extract with a relative density of 1.26 at 50 - 60 °C.

[0196] F. Take the extract obtained in step E, perform belt vacuum drying under the conditions of a vacuum degree of -0.08 MPa - -0.10 Mpa and a drying temperature of 63 °C, pulverize it into fine powder, sieve it, add the β-cyclodextrin inclusion complex obtained in step B, mix well to obtain the fine powder of Jingfang extract. Add the formulated amount of sucrose powder and dextrin (weight ratio 5:2), mix well, make into granules, dry, size the granules, and make 10 kg to obtain the product.

Claims

1. Use of Jingfang preparation in the preparation of a drug for treating sequelae of COVID-19, wherein the sequelae of COVID-19 is acute liver injury.

2. The application according to claim 1, wherein The Jingfang preparation is one of granules, tablets, capsules and microcapsules.

3. The application according to claim 2, wherein The oral pharmaceutical preparation is made from the following traditional Chinese medicine components: Schizonepeta tenuifolia Briq. 97 parts by weight Saposhnikovia divaricata (Turcz.) Schischk. 97 parts by weight Notopterygium incisum Ting ex Hsiao & K. C. Hsia 97 parts by weight Angelica pubescens Maxim. f. biserrata Shan & Yuan 97 parts by weight Bupleurum chinense DC. 97 parts by weight Peucedanum praeruptorum Dunn 97 parts by weight Ligusticum chuanxiong Hort. 97 parts by weight Aurantii Fructus Immaturus 97 parts by weight Poria cocos (Schw.) Wolf 97 parts by weight Platycodon grandiflorus (Jacq.) A. DC. 97 parts by weight Glycyrrhiza uralensis Fisch. 32.4 parts by weight.

4. The application according to claim 2 or 3, characterized in that, The preparation method of the oral pharmaceutical preparation comprises the following steps: A. Schizonepeta tenuifolia Briq., Saposhnikovia divaricata (Turcz.) Schischk., Notopterygium incisum Ting ex Hsiao & K. C. Hsia, Angelica pubescens Maxim. f. biserrata Shan & Yuan, Peucedanum praeruptorum Dunn, Ligusticum chuanxiong Hort. and Aurantii Fructus Immaturus, 7 kinds of medicinal materials are respectively extracted to obtain volatile oils, and 7 kinds of volatile oils of medicinal materials are obtained respectively. The residues after distillation and the aqueous solutions after distillation of Ligusticum chuanxiong Hort. and Aurantii Fructus Immaturus are reserved for later use; B. Take the volatile oils obtained in step A, and respectively add β-cyclodextrin to make inclusion compounds; C. Prepare a 20-30% ethanol solution as a solvent with the aqueous solutions after distillation of Ligusticum chuanxiong Hort. and Aurantii Fructus Immaturus obtained in step A, and percolate the residues of Ligusticum chuanxiong Hort., Aurantii Fructus Immaturus and Poria cocos obtained in step A to obtain a percolate for later use; D. Decoct the residues of Schizonepeta tenuifolia Briq., Saposhnikovia divaricata (Turcz.) Schischk., Notopterygium incisum Ting ex Hsiao & K. C. Hsia, Angelica pubescens Maxim. f. biserrata Shan & Yuan and Peucedanum praeruptorum Dunn obtained in step A together with the remaining three kinds of medicinal materials, Bupleurum chinense DC., Platycodon grandiflorus (Jacq.) A. DC. and Glycyrrhiza uralensis Fisch., 2-3 times with water, each time for 1.5-2.5 hours, combine the decoction liquids, filter, and concentrate the filtrate to an extract with a relative density of 1.18-1.25 at 60-70 °C; E. Combine the percolate obtained in step C and the extract obtained in step D, mix well, let stand, filter, and concentrate the filtrate to an extract with a relative density of 1.22-1.28 at 50-60 °C; F. Take the extract obtained in step E and the β-cyclodextrin inclusion compound obtained in step B, and directly or add pharmaceutically acceptable excipients through conventional processes to make an oral pharmaceutical preparation.

5. The application according to claim 4, wherein The preparation method of the granules comprises the following steps: A. Schizonepeta tenuifolia Briq., Saposhnikovia divaricata (Turcz.) Schischk., Notopterygium incisum Ting ex Hsiao & K. C. Hsia, Angelica pubescens Maxim. f. biserrata Shan & Yuan, Peucedanum praeruptorum Dunn, Ligusticum chuanxiong Hort. and Aurantii Fructus Immaturus, 7 kinds of medicinal materials are respectively extracted to obtain volatile oils, and 7 kinds of volatile oils of medicinal materials are obtained respectively. The residues after distillation and the aqueous solutions after distillation of Ligusticum chuanxiong Hort. and Aurantii Fructus Immaturus are reserved for later use; B. Take the volatile oils obtained in step A, and respectively add β-cyclodextrin to make inclusion compounds; C. Prepare a 25% ethanol solution as a solvent with the aqueous solutions after distillation of Ligusticum chuanxiong Hort. and Aurantii Fructus Immaturus obtained in step A, and percolate the residues of Ligusticum chuanxiong Hort., Aurantii Fructus Immaturus and Poria cocos obtained in step A to obtain a percolate for later use; D. Decoct the residues of Schizonepeta tenuifolia Briq., Saposhnikovia divaricata (Turcz.) Schischk., Notopterygium incisum Ting ex Hsiao & K. C. Hsia, Angelica pubescens Maxim. f. biserrata Shan & Yuan and Peucedanum praeruptorum Dunn obtained in step A together with the remaining three kinds of medicinal materials, Bupleurum chinense DC., Platycodon grandiflorus (Jacq.) A. DC. and Glycyrrhiza uralensis Fisch., 3 times with water, each time for 2 hours, combine the decoction liquids, filter, and concentrate the filtrate to an extract with a relative density of 1.23 at 60-70 °C; E. Combine the percolate obtained in step C and the extract obtained in step D, mix well, let stand, filter, and concentrate the filtrate to an extract with a relative density of 1.26 at 50-60 °C; F. Take the extract obtained in step E, and perform belt vacuum drying under the conditions of a vacuum degree of -0.08 MPa to -0.10 MPa and a drying temperature of 63°C. Crush it into fine powder, sieve it, add the β-cyclodextrin clathrate obtained in step B, mix well, and you will get the fine powder of Jingfang extract. Add the excipients with a weight ratio of sucrose powder: hydroxypropyl starch: mannitol = 3:2:0.5 according to the formula amount, mix well, make granules, dry them, and size them to obtain the product.

6. The application according to claim 4, wherein The preparation method of the microcapsules includes the following steps: A. Extract the volatile oils from 7 herbs including Schizonepeta tenuifolia Briq., Saposhnikovia divaricata (Turcz.) Schischk., Notopterygium incisum Ting ex H. T. Chang, Angelica pubescens Maxim. f. biserrata Shan et Yuan, Peucedanum praeruptorum Dunn, Ligusticum chuanxiong Hort. and Aurantii Fructus Immaturus respectively to obtain 7 kinds of volatile oils from the herbs. Reserve the residues after distillation and the aqueous solutions after distillation of Ligusticum chuanxiong Hort. and Aurantii Fructus Immaturus. B. Take the volatile oils obtained in step A and prepare clathrates with β-cyclodextrin respectively. C. Prepare a 28% ethanol solution with the aqueous solutions after distillation of Ligusticum chuanxiong Hort. and Aurantii Fructus Immaturus obtained in step A as the solvent, and perform percolation on the residues of Ligusticum chuanxiong Hort., Aurantii Fructus Immaturus and Poria cocos obtained in step A to obtain the percolate for standby. D. Decoct the residues of Schizonepeta tenuifolia Briq., Saposhnikovia divaricata (Turcz.) Schischk., Notopterygium incisum Ting ex H. T. Chang, Angelica pubescens Maxim. f. biserrata Shan et Yuan and Peucedanum praeruptorum Dunn obtained in step A together with the other three herbs including Bupleurum chinense DC., Platycodon grandiflorum (Jacq.) A. DC. and Glycyrrhiza uralensis Fisch. twice with water for 2.5 hours each time. Combine the decoction liquids, filter, and concentrate the filtrate to an extract with a relative density of 1.22 at 60 - 70°C. E. Combine the percolate obtained in step C and the extract obtained in step D, mix well, let it stand, filter, and concentrate the filtrate to an extract with a relative density of 1.25 at 50 - 60°C. F. Take the extract obtained in step E, add the cyclodextrin clathrate obtained in step B, mix well, make granules, perform belt vacuum drying, and crush to obtain the fine powder of Jingfang extract for standby. G. Weigh the fine powder of Jingfang extract, capsule material, anti-adhesive agent and plasticizer according to the formula amount. Add the capsule material, anti-adhesive agent and plasticizer to purified water, heat and stir at 54°C to dissolve them, and prepare a capsule material solution with a mass fraction of 33%. Cool it to room temperature, add the fine powder of Jingfang extract and emulsifier under stirring, and perform homogeneous emulsification to obtain an emulsion for standby. H. Spray-dry the emulsion in step G under the conditions of an inlet air temperature of 165°C, a spray pressure of 0.40 MPa, and a feeding rate of 21.5 ml / min. Collect the microcapsules and cool them to obtain the product.

7. The application according to claim 6, wherein The conditions for belt vacuum drying in step F are a vacuum degree of -0.08 MPa to -0.10 MPa and a drying temperature of 56°C.

8. The application according to claim 6, characterized in that The capsule material in step G is sodium caseinate: maltodextrin = 3:2, the anti-adhesive agent is stearyl alcohol: titanium dioxide = 3:1, and the plasticizer is polyethylene glycol: citric acid = 3:

1.

9. The application according to claim 6, wherein The emulsifier in step C is a composite emulsifier of sucrose fatty acid ester: soybean phospholipid = 8:5 by weight, and the dosage is 1.18% of the total amount of the preparation formula by mass fraction.

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