CN114288306B - Use of composition in preparing medicine for inhibiting SARS-CoV-2 virus replication - Google Patents
Use of composition in preparing medicine for inhibiting SARS-CoV-2 virus replication Download PDFInfo
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- CN114288306B CN114288306B CN202210103516.3A CN202210103516A CN114288306B CN 114288306 B CN114288306 B CN 114288306B CN 202210103516 A CN202210103516 A CN 202210103516A CN 114288306 B CN114288306 B CN 114288306B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/58—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin
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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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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The application relates to the field of medicines, in particular to application of a composition in preparing medicines for resisting bunyavirus or SARS-CoV-2 virus. The compositions provided herein are effective against bunyaviruses and against SARS-CoV-2 virus.
Description
The application relates to application of toosendanin in preparation of medicines for preventing and/or treating bunyavirus and novel coronavirus infection diseases, and divisional application of patent application with application number of 202110821896.X, which are submitted by 20 months of 2021 and 20 months of application.
Technical Field
The application relates to the field of medicines, in particular to application of a composition in preparing medicines for resisting bunyavirus or SARS-CoV-2 virus.
Background
Toosendanin (TSN) is a tetracyclic triterpene compound from cortex Meliae (Melia toosendan Sieb. Et Zucc.) and is a main medicinal component of Meliaceae plants.
The structural formula of toosendanin is as follows:
toosendanin has various pharmacological activities, such as anti-botulinum neurotoxin, specific to various ion channels of nerve and cardiomyocytes and inhibiting insect feeding and larval development. In recent years, toosendanin has been reported to induce death and apoptosis of tumor cells (such as liver cancer cells, nerve tumor cells and lymphoma cells), which suggests that toosendanin has good prospect in inhibiting proliferation of tumor cells.
The application provides a new application of toosendanin.
Disclosure of Invention
The embodiment of the application aims to provide an application of a composition in preparing a medicine for resisting bunyavirus or SARS-CoV-2 virus, which aims to provide a new application of toosendanin.
The application provides an application of a composition in preparing a bunyavirus-resistant medicament, wherein the composition comprises toosendanin or pharmaceutically acceptable salt, solvate or hydrate thereof.
In some embodiments of the application, the use of a composition in the manufacture of a medicament for the treatment of fever with thrombocytopenia syndrome virus.
In some embodiments of the application, the use of a composition in the manufacture of a medicament for inhibiting the transmission of fever with thrombocytopenia syndrome virus.
In some embodiments of the application, the use of a composition in the manufacture of a medicament against a rift valley fever virus.
In some embodiments of the application, the use of a composition in the manufacture of a medicament for inhibiting the transmission of anti-split valley fever virus.
In some embodiments of the application, the composition further comprises a pharmaceutically acceptable adjuvant.
The application also provides an application of the composition in preparing anti-SARS-CoV-2 virus medicines, wherein the composition comprises toosendanin or pharmaceutically acceptable salts, solvates or hydrates thereof.
In some embodiments of the application, the use of the composition in the manufacture of a medicament for inhibiting SARS-CoV-2 viral replication.
In some embodiments of the application, the composition further comprises a pharmaceutically acceptable adjuvant.
In some embodiments of the application, the dosage form of the composition is that of the drug, and the dosage form is that of a gastrointestinal administration or injection.
The application of the composition provided by the embodiment of the application in preparing the anti-bunyavirus or SARS-CoV-2 virus medicament has at least the following beneficial effects:
toosendanin or its pharmaceutically acceptable salts, solvates or hydrates are effective against bunyavirus and SARS-CoV-2 virus; further, toosendanin or its pharmaceutically acceptable salts, solvates or hydrates are effective against fever with thrombocytopenia syndrome virus, and anti-split valley fever virus.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings that are needed in the embodiments will be briefly described below, it being understood that the following drawings only illustrate some embodiments of the present application and therefore should not be considered as limiting the scope, and other related drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 shows the cytotoxicity test results of Toosendanin in Experimental example 1.
FIG. 2 shows the results of the antiviral activity assay of Toosendanin at various concentrations in Experimental example 2.
FIG. 3 shows the effect of Toosendanin on SFTSV replication in Experimental example 2.
Figure 4 shows the effect of tosendanine on RVFV replication in experimental example 3.
FIG. 5 shows the effect of Toosendanin on SFTSV infected C57BL/6 mice in experimental example 4.
FIG. 6 shows the effect of Toosendanin on SARS-CoV-2 replication in experimental example 5.
The vehicle group in fig. 2, 4 and 5 refers to the vehicle control group.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present application more clear, the technical solutions of the embodiments of the present application will be clearly and completely described below. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
The use of the composition according to the present application in the preparation of a medicament against bunyavirus or SARS-CoV-2 virus is described in detail below.
In a first aspect, the application provides a use of: use of a composition comprising toosendanin or a pharmaceutically acceptable salt, solvate or hydrate thereof in the manufacture of a medicament against bunyavirus.
Bunyavirus (bunyavir) is a 1-mesh virus with spherical, enveloped and segmented negative-strand RNA.
For example, the use of the above composition in the preparation of a medicament against fever with thrombocytopenia syndrome virus.
Fever with thrombocytopenia syndrome virus (Severe Fever with Thrombocytopenia Syndrome Virus, SFTSV) belongs to the order bunyaviridae (Bunyavirales), the family of Phenuiviridae, genus Bandavirus, is a novel Bunyavirus (Bunyavirus).
In some embodiments of the application, the use in the preparation of a medicament for inhibiting transmission of febrile with thrombocytopenia syndrome virus.
Ticks are a known Natural host (Natural host) for SFTSVs, and tick stings are the main cause of infection of humans with SFTSVs. Toosendanin or pharmaceutically acceptable salts, solvates or hydrates thereof can inhibit the transmission of fever with thrombocytopenia syndrome virus; therefore, has the effects of prevention and treatment.
For example, the use of the above composition in the preparation of a medicament against a split valley fever virus. Setaria virus (Rift Valley fever virus, RVFV) belongs to the family Phenuiviridae, genus Phlebovirus, a highly pathogenic bunyavirus. RVFV is transmitted by mosquito vectors and can infect animals and humans.
Toosendanin or its pharmaceutically acceptable salts, solvates or hydrates can be used for preparing medicines for inhibiting transmission of anti-crack valley fever virus.
In some embodiments, the above composition further comprises a pharmaceutically acceptable adjuvant.
In a second aspect, the application also provides a use of: use of a composition comprising toosendanin or a pharmaceutically acceptable salt, solvate or hydrate thereof for the manufacture of a medicament against SARS-CoV-2 virus.
The novel coronavirus (SARS-CoV-2) belongs to the family Coronaviridae (Coronaviridae), genus beta coronavirus (Betacorovirus).
In an embodiment of the present application, toosendanin or a pharmaceutically acceptable salt, solvate or hydrate thereof has therapeutic effect on novel coronavirus (SARS-CoV-2).
In some embodiments of the application, toosendanin or a pharmaceutically acceptable salt, solvate or hydrate thereof may inhibit SARS-CoV-2 virus replication; thus, the use of the composition in the manufacture of a medicament for inhibiting SARS-CoV-2 viral replication.
Further, the composition also comprises pharmaceutically acceptable auxiliary materials.
In the above-described first and second aspects, the pharmaceutical may be in the form of a tablet, capsule, granule, pill, powder, paste, powder or oral liquid.
As an exemplary, the pharmaceutically acceptable excipients are one or more of water, ethanol, starch, sugar powder, dextrin, lactose, compressible starch, microcrystalline cellulose, inorganic salts, mannitol, starch slurry, sodium carboxymethyl cellulose, methyl cellulose, ethyl cellulose, hydroxypropyl methylcellulose, sodium carboxymethyl cellulose, CMS-Na, L-HPC, crosslinked PVP, CCNa, magnesium stearate, micro powder silica gel, talc, hydrogenated vegetable oil, PEG, magnesium lauryl sulfate, lactose, corn starch, calcium carbonate, calcium phosphate, sodium dodecyl sulfate, tween-80, boric acid, stearic acid, sodium chloride, sodium benzoate, sodium acetate, polyethylene glycol, liquid paraffin, calcium hydrogen phosphate, calcium dihydrogen phosphate, lactose, pregelatinized starch, povidone, citric acid, polysorbate 80, paraffin, hydrogenated vegetable oil, glycine, CAP, AEA, acrylic resin, cyclodextrin, acacia, gelatin, and sodium alginate.
The subject to which the pharmaceutical compositions of the present application are administered is not limited to humans, but may be, for example, other mammals. Any method of administration may be administered, for example, orally or non-orally. Parenteral methods of administration may be intravenous, arterial, bone marrow, dura mater, heart, transdermal, subcutaneous, luminal, nasal, intestinal, topical, sublingual, or rectal.
Further, pharmaceutically acceptable excipients may be selected according to the dosage form.
It should be further noted that, in addition to the auxiliary materials, other pharmaceutically acceptable active ingredients may be included in the composition of the present application.
Alternatively, the preparation method based on toosendanin can also comprise pharmaceutically acceptable impurities.
For example, the composition may be an extract of Melia azedarach (Melia toosendan).
The term "pharmaceutically acceptable" refers to a nontoxic composition which is physiologically acceptable and does not inhibit the action of the active ingredient after administration to a human, and does not usually cause allergic reactions such as gastrointestinal disorders and vertigo or the like.
The pharmaceutically acceptable salts are preferably pharmaceutically acceptable acid addition salts formed with free acid. The free acid is an organic acid and an inorganic acid which can be used all together. Furthermore, in addition to the organic acid, citric acid, acetic acid, lactic acid, tartaric acid, maleic acid, fumaric acid, formic acid, propionic acid, oxalic acid, trifluoroacetic acid, benzoic acid, gluconic acid, methanesulfonic acid, glycolic acid, succinic acid, 4-toluenesulfonic acid, carbonic acid and aspartic acid can be used. The inorganic acid may be hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid.
The features and properties of the present application are described in further detail below with reference to experimental examples and examples.
Experimental example 1
Cytotoxicity detection of Toosendanin (Toosendanin)
In HUVECs cells, the cytotoxicity of Toosendanin was examined. HUVECs cells were cultured at 1X 10 4 Cells/well were seeded in 96-well plate cell culture plates at 37℃with 5% CO 2 Cultures were incubated in the incubator for 12-16h, treated with 0.2. Mu.M, 0.62. Mu.M, 1.85. Mu.M, 5.6. Mu.M, 16.7. Mu.M, 50. Mu.M, 150. Mu.M and 450. Mu.M Toosendanin, three duplicate wells per group, and the control group was added with the same amount of dimethyl sulfoxide (DMSO). After 24h of drug action, 10 μl of CCK8 was added, incubated at 37deg.C for 1h, and OD was detected 450nm Analyzing the cell activity; the detection results are shown in FIG. 1.
Wherein CCK8 is a kit for cell activity assay comprising in solutionIs a water-soluble tetrazolium salt, which can be oxidized and reduced into orange-yellow water-soluble Formazan (Formazan) by intracellular dehydrogenase, and the generation amount of the Formazan is in direct proportion to the number of living cells.
Tested as shown in FIG. 1, toosendanin's CC 50 Greater than 450 μm. In the subsequent examples, toosendanin was used at a concentration of 1-40. Mu.M, within a safe and nontoxic range.
Experimental example 2
Detection of antiviral Activity of Toosendanin (Toosendanin) on SFTSV
Vero cells according to 2X 10 5 Cells/well were seeded in 24-well plate cell culture plates at 37℃with 5% CO 2 The cells were incubated in the incubator for 12-16h, treated with Toosendanin at concentrations of 5. Mu.M, 10. Mu.M, 20. Mu.M, 40. Mu.M, and the negative control was added with the same volume of DMSO. After 1h incubation, infection with SFTSV, 2h incubation at 37℃and replacement of DMEM medium with 2% FBS, 24h later, the effect of Toosendanin on inhibiting viral replication was examined by titer, and as shown in FIG. 2, the compound Toosendanin showed significant inhibition of SFTSV replication at different concentrations. IC for further inhibiting SFTSV in Vero cells by Toosendanin 50 Detection is carried out, and the inhibition curve of the SFTSV is shown in figure 3; the circles in FIG. 3 represent the inhibition rate of the drug to inhibit viral replication, the triangles represent cytotoxicity, and as can be seen from FIG. 3, tosendanin significantly inhibited SFTSV activity, calculated as IC 50 1.2. Mu.M. CC in Vero cells 50 Greater than 250 mu M, SI index greater than 208.3。
The toosendanin has the function of inhibiting SFTSV activity, and can be used for preparing medicines for resisting fever with thrombocytopenia syndrome virus.
Experimental example 3
Effect of Toosendanin (Toosendanin) on RVFV replication
HUVECs cells were cultured at 2X 10 5 Cells/well were seeded in 24-well plate cell culture plates at 37℃with 5% CO 2 Cultures were performed in incubators for 12-16h at concentrations of 0.2. Mu.M, 0.62. Mu.M, 1.85. Mu.M, 5.6. Mu.M, 16.7. Mu.M, 50. Mu.M, and the negative control was added to the same volume of DMSO. After 1h incubation, infection with Rift Valley Fever Virus (RVFV) belonging to the genus sand fly virus, after 2h incubation at 37 ℃, DMEM medium with 2% fbs was replaced, and after 24h, viral gene copy number in the supernatant was detected by real-time fluorescent quantitative PCR (qPCR) method, as shown in fig. 4; as can be seen from fig. 4, the compound tosendanin has a significant inhibitory effect on replication of the Rift Valley Fever Virus (RVFV) at different concentrations; indicating that toosendanin has the function of inhibiting the replication of the valley fever virus.
Experimental example 4
Effect of Toosendanin (Toosendanin) on SFTSV infected C57BL/6 mice
The immune system of the C57BL/6 mouse is sound, the genetic background is clean, and the C57BL/6 mouse can be used for drug protection evaluation experiments. The mice were all kept in SPF environment and the mice were divided into 2 groups: (1) Sftsv+veccle group (n=5), (2) sftsv+tosendanin group (n=5). The mice were infected by intraperitoneal injection with a toxin receiving amount of 100. Mu.L (10 5 FFU/only), mock group was an equal volume of DMEM medium. The dosage of Toosendanin was 1mg/kg/d by intraperitoneal injection, and the administration was started 3 days before infection, 1 time a day for 6 days. The whole experimental procedure strictly complies with guidelines of the national institutes of health, as set forth in the guidelines of the operation of the institutes of animal protection and use. The daily monitoring indicators of the mice included: bow back, hair standing, motility, stress, weight, etc. The mice were dissected on day 3 of SFTSV infection and spleens were taken to test their viral load. The results are shown in FIG. 5, where the spleen virus titer of mice in the administered group was significantly reduced compared to the control group. It was suggested that Toosendanin could be displayed at the mouse levelInhibiting replication of the SFTSV.
TSN in fig. 5 represents the sftsv+tosendanin group.
Experimental example 5
Effect of Toosendanin (Toosendanin) on SARS-CoV-2 replication
IC for inhibiting SARS-CoV-2 by Toosendanin in Vero-E6 cell 50 And CC 50 The results of the detection are shown in the inhibition curve of SARS-CoV-2 in FIG. 6, the square block in FIG. 6 shows the inhibition ratio of the drug to inhibit virus replication, the round dot shows cytotoxicity, and Toosendanin significantly inhibits the activity of SARS-CoV-2, and its IC is calculated 50 0.24. Mu.M. CC in Vero-E6 cells 50 Greater than 450 μm, SI index greater than 1875.
The toosendanin can inhibit SARS-CoV-2 virus replication, and can be used for preparing anti-SARS-CoV-2 virus medicine.
In the above experimental examples 1-5, the present application provides a pharmaceutical composition comprising toosendanin or a pharmaceutically acceptable salt, solvate or hydrate thereof.
The medicine can be used for resisting bunyavirus, fever with thrombocytopenia syndrome virus, anti-crack-resistant valley fever virus and SARS-CoV-2 virus.
Taken together, it can be seen that toosendanin or its pharmaceutically acceptable salts, solvates or hydrates are effective against bunyavirus, fever with thrombocytopenia syndrome virus, anti-split valley fever virus and SARS-CoV-2 virus.
The above description is only of the preferred embodiments of the present application and is not intended to limit the present application, but various modifications and variations can be made to the present application by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the protection scope of the present application.
Claims (3)
1. The application of a composition in preparing a medicine for inhibiting SARS-CoV-2 virus replication is characterized in that the active ingredient of the composition is toosendanin or pharmaceutically acceptable salts thereof.
2. The use of claim 1, the composition further comprising a pharmaceutically acceptable adjuvant.
3. The use according to claim 1, wherein the composition is in the form of a gastrointestinal tract administration or injection.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202210103516.3A CN114288306B (en) | 2021-07-20 | 2021-07-20 | Use of composition in preparing medicine for inhibiting SARS-CoV-2 virus replication |
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| CN202110821896.XA CN113350361B (en) | 2021-07-20 | 2021-07-20 | Application of composition in preparation of anti-bunyavirus or SARS-CoV-2 virus medicine |
| CN202210103516.3A CN114288306B (en) | 2021-07-20 | 2021-07-20 | Use of composition in preparing medicine for inhibiting SARS-CoV-2 virus replication |
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| CN202110821896.XA Division CN113350361B (en) | 2021-07-20 | 2021-07-20 | Application of composition in preparation of anti-bunyavirus or SARS-CoV-2 virus medicine |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101991583A (en) * | 2009-08-25 | 2011-03-30 | 财团法人工业技术研究院 | Anti-hepatitis C composition and method for preparing drugs for inhibiting hepatitis C virus or treating hepatitis C |
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| CN1548146A (en) * | 2003-05-20 | 2004-11-24 | 刘树林 | Chinese medicine for preventing and treating virus disease |
| US8802662B2 (en) * | 2009-06-16 | 2014-08-12 | Industrial Technology Research Institute | Anti-hepatitis C composition and method for preparing drug for inhibiting hepatitis C viruses or treating hepatitis C |
| CN103450324A (en) * | 2013-09-17 | 2013-12-18 | 南京通泽农业科技有限公司 | Method for extracting toosendanin from cortex meliae |
| CN111166862A (en) * | 2020-02-24 | 2020-05-19 | 陈文才 | Laggera pterodonta composition for treating new coronary pneumonia and application thereof |
| CN111449093A (en) * | 2020-05-20 | 2020-07-28 | 商磊 | Natural essential oil composition with antibacterial and disinfecting effects and application thereof |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101991583A (en) * | 2009-08-25 | 2011-03-30 | 财团法人工业技术研究院 | Anti-hepatitis C composition and method for preparing drugs for inhibiting hepatitis C virus or treating hepatitis C |
Non-Patent Citations (1)
| Title |
|---|
| Young-Hee Jin ；等.Toosendanin From Melia Fructus Suppresses Influenza A Virus Infection by Altering Nuclear Localization of Viral Polymerase PA Protein.《Front Pharmacol》.2019,(第10期),1025. * |
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| CN113350361A (en) | 2021-09-07 |
| CN113350361B (en) | 2022-03-08 |
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