CN111150755B - Traditional Chinese medicine composition for preventing and treating viral diseases and application thereof - Google Patents
Traditional Chinese medicine composition for preventing and treating viral diseases and application thereof Download PDFInfo
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
- A61K36/532—Agastache, e.g. giant hyssop
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
- A61K36/282—Artemisia, e.g. wormwood or sagebrush
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
- A61K36/287—Chrysanthemum, e.g. daisy
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
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- A—HUMAN NECESSITIES
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P13/00—Drugs for disorders of the urinary system
- A61P13/12—Drugs for disorders of the urinary system of the kidneys
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/37—Extraction at elevated pressure or temperature, e.g. pressurized solvent extraction [PSE], supercritical carbon dioxide extraction or subcritical water extraction
Abstract
The invention discloses a new application of a traditional Chinese medicine composition in preparing a medicament for preventing and treating viral diseases. The invention carries out the research of the drug effect of the traditional Chinese medicine composition on resisting the common coronavirus and the novel coronavirus in vitro through experiments, and the result shows that the traditional Chinese medicine composition has certain inhibiting effect on the common coronavirus HcoV-229E and the novel coronavirus SARS-Cov-2 virus. The Chinese medicinal composition can inhibit the expression of cytokines such as TNF-alpha, IL-6 and the like generated by HcoV-229E, SARS-Cov-2 virus induction. Has certain therapeutic effect on inflammatory diseases induced by HcoV-229E, SARS-Cov-2 virus, such as pneumonia, nephritis, hepatitis and the like, and has good clinical application prospect.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicine preparations, and particularly relates to an application of a traditional Chinese medicine for treating viral diseases.
Background
Human coronavirus HCoV-229E belongs to alpha group positive strand RNA virus coronavirus, is a pathogen causing upper respiratory tract infection of human, and often causes common cold of human. Clinically, it causes gastrointestinal diseases and nervous system symptoms in addition to respiratory tract infection. It is reported that type I interferon alpha (IFN-alpha) has certain therapeutic effect on HCoV-229E, but IFN-alpha has certain adverse reaction.
Symptoms of pneumonia infection caused by novel coronavirus (SARS-CoV-2) comprise general symptoms, severe symptoms and atypical symptoms, wherein the general symptoms mainly comprise fever, hypodynamia, dry cough and gradual dyspnea, the severe symptoms comprise coagulation dysfunction, uncorrectable metabolic acidosis, septic shock, acute respiratory distress syndrome and the like, the atypical symptoms comprise anorexia and hypodynamia, mental retardation, nausea and vomiting, diarrhea and the like which take a digestive system as first symptoms, headache which takes nervous system symptoms as first symptoms, conjunctivitis which takes ophthalmic symptoms as first symptoms, cardiovascular symptoms, chest distress and palpitation which take only mild muscle soreness of limbs or waist and back.
The novel coronavirus is a highly infectious virus, and the transmission route is mainly through respiratory droplet transmission, contact transmission and the like. At present, no effective treatment medicine for the novel coronavirus SARS-CoV-2 is available on the market, which brings great harm to the life health of patients.
The traditional Chinese medicine has unique advantages in the aspect of antivirus, so that the research and the screening of the traditional Chinese medicine preparation with antivirus are necessary on the basis of the prior art, and the clinical indications of the traditional Chinese medicine preparation are enlarged.
Disclosure of Invention
The purpose of the invention is as follows: the invention aims to exert the advantages of the traditional Chinese medicine, screen out the traditional Chinese medicine preparation with antiviral effect, expand the clinical indications of the traditional Chinese medicine preparation, and provide the traditional Chinese medicine preparation with good prevention and treatment effects on coronavirus.
The technical scheme is as follows: in order to achieve the above purpose, the invention adopts the technical scheme that:
the application of the Chinese medicinal composition in preparing medicine for preventing and treating viral diseases is provided.
Application of the Chinese medicinal composition in preparing medicine for preventing and treating coronavirus diseases is provided.
The application of the Chinese medicinal composition in preparing medicine for preventing and treating common coronavirus HcoV-229E or novel coronavirus SARS-Cov-2 virus diseases is provided.
The application of the Chinese medicinal composition in preparing medicine for preventing and treating inflammation diseases caused by common coronavirus HcoV-229E and novel coronavirus SARS-Cov-2 virus is provided. Preferably, the inflammatory disease includes pneumonia, nephritis, hepatitis, gastroenteritis or neuritis.
In the application of the invention, the traditional Chinese medicine composition comprises wrinkled gianthyssop herb, wild chrysanthemum flower, Chinese mosla herb and sweet wormwood herb.
Preferably, the traditional Chinese medicine composition comprises 10-50 parts of agastache rugosus, 10-50 parts of wild chrysanthemum flower, 10-30 parts of elsholtzia and 10-50 parts of artemisia apiacea.
In a particularly preferred scheme, the traditional Chinese medicine composition comprises 40 parts of wrinkled gianthyssop herb, 40 parts of wild chrysanthemum flower, 24 parts of Chinese mosla herb and 40 parts of sweet wormwood herb.
The preparation method of the traditional Chinese medicine composition comprises the following steps: taking the agastache rugosus, the wild chrysanthemum, the elsholtzia and the sweet wormwood according to the weight proportion, extracting volatile oil by a steam distillation method, and storing in another container. Decocting the medicine residues with water (preferably decocting for 1-3 times and 1-2 hours each time), concentrating the decoction, standing for precipitation, filtering, and concentrating the filtrate into clear paste. Adding sucrose into the fluid extract, granulating, drying, adding the above volatile oil and oleum Menthae Dementholatum, mixing, adding pharmaceutically acceptable vehicle, and making into tablet, capsule, pill, oral liquid or mixture.
The xiangju cold treating granule is produced by leiying pharmaceutical industry group limited company and has the Chinese medicine standard Z20003016 and the medicinal ingredients comprise 40 parts of wrinkled gianthyssop, 40 parts of wild chrysanthemum flower, 24 parts of elsholtzia and 40 parts of sweet wormwood herb.
Has the advantages that: the invention carries out the in vitro safety of the traditional Chinese medicine composition and the pharmacodynamic study of the antibody in vitro common coronavirus and the novel coronavirus through experiments, and the result shows that the traditional Chinese medicine composition has certain inhibiting effect on the common coronavirus HcoV-229E and the novel coronavirus SARS-Cov-2 virus.
In addition, the invention carries out in-vitro infection induced inflammation experiments on the common coronavirus HcoV-229E and the novel coronavirus SARS-Cov-2 by the traditional Chinese medicine composition through experiments, and the result shows that after the HcoV-229E, SARS-Cov-2 virus is induced and intervened in HUH7 cells, the expression of cytokines such as TNF-alpha, IL-6, CCL-2/MCP-1, CXCL-10/IP-10 and the like in a virus group is obviously improved, and after the intervention of the traditional Chinese medicine composition is carried out for 24 hours, the concentration of each medicine has inhibition effect of different degrees, thereby confirming that the traditional Chinese medicine composition has the effect of obviously inhibiting the expression of the cytokines such as TNF-alpha, IL-6 and the like induced and generated by the HcoV-229E, SARS-Cov-2 virus. Has good therapeutic effect on various inflammatory diseases (such as pneumonia, nephritis, gastroenteritis, hepatitis, neuritis and the like) induced by HcoV-229E, SARS-Cov-2 virus, and has good clinical application prospect.
Drawings
FIG. 1 is a bar graph of the effect of a Chinese medicinal composition on HcoV-229E virus-induced inflammatory factors in vitro.
FIG. 2 is a bar graph of the effect of the Chinese medicinal composition on SARS-Cov-2 virus-induced inflammatory factors in vitro.
In fig. 1 and 2, the statistical significance (P < 0.01) and the statistical significance (P < 0.05) were shown in comparison with the model group.
Detailed Description
The present invention is further illustrated by the following examples, which are intended to be purely exemplary and are not intended to limit the scope of the invention, as various equivalent modifications of the invention will occur to those skilled in the art upon reading the present disclosure and fall within the scope of the appended claims.
EXAMPLE 1 preparation of Chinese medicinal preparation
Taking 40 parts of wrinkled gianthyssop herb, 40 parts of wild chrysanthemum flower, 24 parts of Chinese mosla herb and 40 parts of sweet wormwood herb according to the weight proportion, adding 8 times of water, extracting for 2 hours by a steam distillation method to obtain volatile oil, and storing in another container. Decocting the residue after steam distillation with 10 times of water for 2 times, each for 2 hr, mixing decoctions, concentrating, standing for precipitation, filtering, and concentrating the filtrate to obtain fluid extract. Adding sucrose into the fluid extract, granulating, drying, adding the above volatile oil and oleum Menthae Dementholatum, and mixing to obtain extract.
Mixing the above extracts with pharmaceutically acceptable carrier, and making into tablet, capsule, pill, oral liquid or mixture.
Example 2 in vitro antiviral Activity test of the Chinese medicinal composition against the common coronavirus HcoV-229E and 2019 novel coronavirus SARS-Cov-2
1. Experimental methods
1.1 VERO E6 cell (African green monkey kidney cell line) toxicity test (MTT method)
The monolayer cells of the 96-well plate were washed 1 time with PBS, the supernatant was discarded, and 100. mu.L of the 2-fold gradient diluted herbal composition was added to each well. Adding equal volume of culture medium into each well of blank control group and normal cell group, at 34 deg.C and 5% CO2Culturing under the condition for 2-5 days. Subsequently 20. mu.L of MTT solution at a concentration of 5mg/mL was added per well and incubation was continued for 4 h. The supernatant was discarded, 100. mu.L of DMSO was added to each well, and the mixture was shaken at a low speed for 5min to completely melt the crystals. And measuring the light absorption value on an enzyme-linked immunosorbent assay (ELISA) at 490nm, and calculating the inhibition rate.
The half-Toxic Concentration (TC) of the drug was calculated by Prism non-regression analysis software50)。
1.2 antiviral efficacy test (MTT method)
The common coronavirus HcoV-229E and the novel coronavirus SARS-Cov-2 are clinical isolates of the first hospital affiliated to Guangzhou medical university.
The monolayer cells on the 96-well plate were washed 1 time with PBS, 100. mu.L/well of 100TCID50 diluted virus (containing 1.5. mu.g/mL TPCK trypsin) was added, and the mixture was incubated at 37 ℃ with 5% CO2Incubating in incubator for 2h, discarding virus solution, adding 2 times of gradient diluted extract of the Chinese medicinal composition of example 1, setting 4 multiple wells for each concentration, taking maximum nontoxic concentration as initial concentration of the medicine, and incubating at 34 deg.C and 5% CO2Incubate in incubator for 2 d. Cytopathic effect (CPE) was recorded daily. The OD was determined by staining with MTT (same as in step 1.1). The effective half concentration (IC) of the drug was calculated using Prism non-regression analysis software50) Calculating selection index SI, SI ═ TC50/IC50). [ refer to "pharmacological Experimental methodology, Xutaiyun Suo Shu (eds.), therapeutic index (SI)>1 denotes effective]。
2. Results of the experiment
2.1, the in vitro safety of the traditional Chinese medicine composition and the pharmacodynamic research results of the antibody against the common coronavirus and the novel coronavirus show that the traditional Chinese medicine composition has obvious inhibiting effect on the common coronavirus HcoV-229E and the novel coronavirus SARS-Cov-2 virus. The half-effective concentration IC50 values and Selection Index (SI) are shown in Table 1 below.
TABLE 1 inhibitory Effect of Chinese medicinal composition on Hcov-229E and SARS-Cov-2 Virus in vitro (MTT method)
The in vitro safety of the Chinese medicinal composition and the inhibition effect of the antibody of the common coronavirus and the novel coronavirus are determined by the antiviral efficacy test (MTT method).
Example 3 pharmaceutical effect of Chinese medicinal composition on inducing inflammation by infection of common coronavirus Hcov-229E and 2019 novel coronavirus SARS-Cov-2 in vitro
1. Experimental methods
1.1, real-time fluorescent quantitative PCR detection of the influence of the traditional Chinese medicine composition on the HcoV-229E, SARS-Cov-2 virus-induced inflammatory factor
1.1.1 preparation of cell samples
HUH7 cells (human liver cancer cells) with good growth state were collected, digested, adjusted for cell density, and inoculated into 6-well plates to obtain a cell density of 4X 105Culturing each cell/mL for 18-24 hr, removing culture medium after the adherent growth is full, washing with PBS for 2 times, adding virus-containing serum-free culture medium, 37 deg.C, and 5% CO2Culturing for 2h, discarding virus solution, washing with PBS for 2 times, adding extracts of the Chinese medicinal composition of example 1 at different concentrations, and testing to obtain three dosage groups of the extract of Chinese medicinal composition, including high dosage group (2 μ g/mL), medium dosage group (1 μ g/mL), and low dosage group (0.5 μ g/mL), virus control group, and normal control group, at 37 deg.C and 5% CO2The culture was continued for 24 h. The cell supernatant was discarded, the cells were washed twice with PBS, 1 mL/sample of Trizol solution was added, and the cells were incubated at room temperature for 5 min.
1.1.2 extraction of RNA
Adding chloroform 200 μ L into cells and cell supernatant respectively, shaking for 15s, incubating at room temperature for 5min, centrifuging at 4 deg.C and 10000rpm for 10 min; after centrifugation, the supernatant was transferred to a new 1.5mL EP tube, 500. mu.L of isopropanol was added, incubation was carried out for 10min,centrifuging at 4 deg.C and 10000rpm for 10 min; centrifuging, adding 1000 μ L75% ethanol, mixing, centrifuging at 4 deg.C and 10000rpm for 10 min; discarding the supernatant, standing at room temperature for 5min, adding RNase-free ddH2Dissolving RNA and performing reverse transcription to obtain cDNA; the total RNA concentration (ng/. mu.L) was determined, the concentration of total RNA extracted was calculated, and the volume of total RNA required was calculated.
1.1.3 reverse transcription of sample RNA
The RNA is subjected to genome-removing DNA reaction before reverse transcription, and the reaction system is shown as the following table:
| reagent | Amount of the composition used |
| 5×gDNA Eraser Buffer | 2.0μL |
| gDNA Eraser | 1.0μL |
| Total RNA | 1.0μL |
| RNase Free dH2O | upto10μL |
Reaction conditions are as follows: 42 ℃ for 2min
Storing at 4 deg.C
Reverse transcription of mRNA into cDNA; the reverse transcription reaction system is configured as the following table:
the reverse transcription reaction conditions were as follows:
37℃ 15min
85℃ 5s
4℃ ∞
1.1.4 real-time fluorescent quantitative PCR detection
The reaction system is as follows:
the PCR reaction conditions are as follows:
1.1.5 data processing
And (3) performing data analysis on the Ct value detected by each group by adopting ABI7500 software, and performing relative quantification by adopting a delta Ct method.
2. Results of the experiment
The real-time fluorescence quantitative PCR detection of the effect of the traditional Chinese medicine composition on the HcoV-229E, SARS-Cov-2 virus-induced inflammatory factor shows the effect of the traditional Chinese medicine composition on the HcoV-229E virus-induced inflammatory factor in vitro in FIG. 1 and the effect of the traditional Chinese medicine composition on the SARS-Cov-2 virus-induced inflammatory factor in vitro in FIG. 2. Experimental results show that after the induction intervention of the HcoV-229E, SARS-Cov-2 virus in HUH7 cells, the expression of cytokines such as TNF-alpha, IL-6, CCL-2/MCP-1, CXCL-10/IP-10 and the like in a virus group is remarkably increased, and after the intervention of the traditional Chinese medicine composition for 24 hours, the traditional Chinese medicine composition at each concentration has the effect of inhibiting the expression of the cytokines at different degrees, so that the traditional Chinese medicine composition is proved to have the effects of inhibiting the expression of the cytokines such as TNF-alpha, IL-6 and the like generated by the induction of the HcoV-229E, SARS-Cov-2 virus and play a role in treating inflammatory diseases such as pneumonia and nephritis and the like caused by the HcoV-229E, SARS-Cov-2 virus.
Of course, the foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (4)
1. The application of a Chinese medicinal composition comprising herba Agastaches, flos Chrysanthemi Indici, herba Moslae and herba Artemisiae Annuae in preparing medicine for preventing and treating severe acute respiratory syndrome coronavirus 2 virus and inflammation diseases caused by the same;
the traditional Chinese medicine composition comprises 10-50 parts of wrinkled gianthyssop herb, 10-50 parts of wild chrysanthemum flower, 10-30 parts of elsholtzia and 10-50 parts of sweet wormwood herb.
2. The use of claim 1, wherein the inflammatory disease comprises a disease selected from the group consisting of pneumonia, nephritis, hepatitis, gastroenteritis, and neuritis.
3. The use of claim 2, wherein the Chinese medicinal composition comprises 40 parts of wrinkled gianthyssop herb, 40 parts of wild chrysanthemum flower, 24 parts of elsholtzia herb and 40 parts of sweet wormwood herb.
4. The use of claim 1, wherein the preparation method of the Chinese medicinal composition comprises:
(1) taking agastache rugosus, wild chrysanthemum, elsholtzia and sweet wormwood according to the weight proportion, extracting volatile oil by a steam distillation method, and collecting the volatile oil;
(2) decocting the residue with water, concentrating the decoction, standing for precipitation, filtering, and concentrating the filtrate to obtain fluid extract;
(3) adding sucrose into the fluid extract obtained in step (2) to prepare granules, drying, adding the volatile oil and the peppermint oil obtained in step (1), mixing uniformly, and preparing the mixture with a pharmaceutically acceptable carrier into tablets, capsules, pills, oral liquid or mixtures.
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Denomination of invention: Traditional Chinese medicine composition for preventing and treating viral diseases and its application Effective date of registration: 20221019 Granted publication date: 20220412 Pledgee: Shanghai Bank Co.,Ltd. Suzhou Branch Pledgor: LEIYUNSHANG PHARMACEUTICAL GROUP Co.,Ltd. Registration number: Y2022320010593 |
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