CN114288306A - Application of composition in preparation of anti-bunyavirus or SARS-CoV-2 virus medicine - Google Patents
Application of composition in preparation of anti-bunyavirus or SARS-CoV-2 virus medicine Download PDFInfo
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- CN114288306A CN114288306A CN202210103516.3A CN202210103516A CN114288306A CN 114288306 A CN114288306 A CN 114288306A CN 202210103516 A CN202210103516 A CN 202210103516A CN 114288306 A CN114288306 A CN 114288306A
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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
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- 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
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Abstract
The application relates to the field of medicines, in particular to application of a composition in preparing a medicine for resisting bunyavirus or SARS-CoV-2 virus. The compositions provided herein are effective against bunyavirus as well as against SARS-CoV-2 virus.
Description
The application is filed on the 20 th 07 th 2021 year, the invention name is the application of toosendanin in preparing the medicine for preventing and/or treating bunyavirus and novel coronavirus infection diseases, and the divisional application of the invention patent application with the application number of 202110821896. X.
Technical Field
The application relates to the field of medicines, in particular to application of a composition in preparing a medicine for resisting bunyavirus or SARS-CoV-2 virus.
Background
Toosendanin (TSN) is a tetracyclic triterpene compound in cortex Meliae (Melia tosendan Sieb. et Zucc.), and is the main medicinal component of Meliaceae plants.
The structural formula of toosendanin is as follows:
toosendanin has various pharmacological activities, such as resisting botulinum neurotoxin, specifically acting on various ion channels of nerve and myocardial cells, and inhibiting insect feeding and larva development. In recent years, toosendanin has been reported to induce the death and apoptosis of tumor cells (such as liver cancer cells, nerve tumor cells and lymph cancer cells), which suggests that toosendanin has a good prospect in inhibiting the proliferation of tumor cells.
The application provides a new application of toosendanin.
Disclosure of Invention
The embodiment of the application aims to provide the application of the composition in preparing a medicine for resisting bunyavirus or SARS-CoV-2 virus, and aims to provide a new application of toosendanin.
The application provides an application of a composition in preparing a medicine for resisting bunyavirus, wherein the composition comprises toosendanin or pharmaceutically acceptable salts, solvates or hydrates thereof.
In some embodiments of the present application, the use of the composition in the preparation of a medicament against fever with thrombocytopenia syndrome virus.
In some embodiments of the present application, the use of a composition in the manufacture of a medicament for inhibiting transmission of a fever with thrombocytopenia syndrome virus.
In some embodiments of the present application, the use of the composition in the preparation of a medicament against rift valley fever.
In some embodiments of the present application, the use of a composition in the manufacture of a medicament for inhibiting the spread of anti-rift valley fever virus.
In some embodiments of the present application, the composition further comprises a pharmaceutically acceptable excipient.
The application also provides an application of a composition in preparing a medicine for resisting SARS-CoV-2 virus, wherein the composition comprises toosendanin or pharmaceutically acceptable salts, solvates or hydrates thereof.
In some embodiments of the present application, the composition is for use in the preparation of a medicament for inhibiting SARS-CoV-2 virus replication.
In some embodiments of the present application, the composition further comprises a pharmaceutically acceptable excipient.
In some embodiments of the present application, the composition is in the form of a gastrointestinal administration form or an injection.
The application of the composition provided by the embodiment of the application in preparing the anti-bunyavirus or SARS-CoV-2 virus medicament at least has the following beneficial effects:
toosendanin or a pharmaceutically acceptable salt, solvate or hydrate thereof is effective against bunyavirus and against SARS-CoV-2 virus; further, toosendanin or a pharmaceutically acceptable salt, solvate or hydrate thereof is effective against fever with thrombocytopenia syndrome virus and anti-rift valley fever virus.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings that are required to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present application and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained from the drawings without inventive effort.
FIG. 1 shows the results of cytotoxicity test of Toosendanin in Experimental example 1.
FIG. 2 shows the results of the measurement of antiviral activity of Toosendanin at different concentrations in Experimental example 2.
FIG. 3 shows the effect of Toosendanin on SFTSV replication in Experimental example 2.
FIG. 4 is a graph showing the effect of Toosendanin on RVFV replication in Experimental example 3.
FIG. 5 is a graph showing the effect of Toosendanin on SFTSV infected C57BL/6 mice in Experimental example 4.
FIG. 6 shows the effect of Toosendanin on the replication of SARS-CoV-2 in Experimental example 5.
The vehicle group in fig. 2, 4 and 5 is referred to as a vehicle control group.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions of the embodiments of the present application will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
The following is a detailed description of the use of the composition of the examples of the present application in the preparation of a medicament against bunyavirus or SARS-CoV-2 virus.
In a first aspect, the present 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 (Bunyavirus) is a 1-mesh virus with spherical, enveloped and segmented negative-strand RNA.
For example, the composition is used for preparing the anti-fever with thrombocytopenia syndrome virus medicine.
The febrile with Thrombocytopenia Syndrome Virus (SFTSV) belongs to the order Bunyavirales (Bunyavirales), the family Pheuiviridae, the genus Bandavirus, and is a novel Bunyavirus (Bunyaviras).
In some embodiments of the present application, use in the preparation of a medicament for inhibiting transmission of a fever with thrombocytopenia syndrome virus.
Ticks are known Natural hosts of SFTSV (Natural host), and tick bites are the major cause of SFTSV infection in humans. Toosendanin or pharmaceutically acceptable salts, solvates or hydrates thereof can inhibit transmission of fever with thrombocytopenia syndrome virus; thus having preventive and therapeutic effects.
For example, the composition is used for preparing a medicament for resisting rift valley fever virus. Rift Valley Fever Virus (RVFV) belongs to the family Phenguiviridae, belongs to the genus Phlebovirus, and is a highly pathogenic bunyavirus. RVFV is transmitted through mosquito vectors and can infect animals and humans.
Toosendanin or pharmaceutically acceptable salts, solvates or hydrates thereof can be used for preparing medicines for inhibiting the transmission of anti-Valley fever virus.
In some embodiments, the composition further comprises a pharmaceutically acceptable excipient.
In a second aspect, the present application also 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 SARS-CoV-2 virus.
The novel coronavirus (SARS-CoV-2) belongs to the family Coronaviridae (Coronaviridae), the genus Betacoronavirus (Betacoronavirus).
In the examples of the present application, azadirachtin or a pharmaceutically acceptable salt, solvate or hydrate thereof has a therapeutic effect on a novel coronavirus (SARS-CoV-2).
In some embodiments of the present application, azadirachtin, or a pharmaceutically acceptable salt, solvate or hydrate thereof, inhibits SARS-CoV-2 virus replication; therefore, the composition is used for preparing the medicine for inhibiting the SARS-CoV-2 virus replication.
Further, the composition also comprises pharmaceutically acceptable auxiliary materials.
In the use of the first and second aspects, the pharmaceutical formulation may be a tablet, capsule, granule, pill, powder, paste, powder, or oral liquid.
Illustratively, the above pharmaceutically acceptable excipients are water, ethanol, starch, powdered sugar, dextrin, lactose, compressible starch, microcrystalline cellulose, inorganic salts, mannitol, starch slurry, sodium carboxymethylcellulose, methylcellulose, ethylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, CMS-Na, L-HPC, crosslinked PVP, CCNa, magnesium stearate, aerosil, talc, hydrogenated vegetable oil, PEG, magnesium lauryl sulfate, lactose, corn starch, calcium carbonate, calcium phosphate, sodium lauryl 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, One or more of cyclodextrin, gum arabic, gelatin, and sodium alginate.
The subject to which the drug prepared by the composition of the present application is administered is not limited to humans, and may be, for example, other mammals. Any method of administration may be, for example, oral or non-oral. Non-oral administration may be intravenous, arterial, bone marrow, dura mater, cardiac, transdermal, subcutaneous, buccal, nasal, enteral, topical, sublingual or rectal.
Further, pharmaceutically acceptable excipients may be selected according to the dosage form.
It is further noted that the composition of the present application may further comprise other pharmaceutically acceptable active ingredients in addition to the adjuvants.
Alternatively, the pharmaceutical composition may further comprise pharmaceutically acceptable impurities based on the preparation method of toosendanin.
For example, the composition may be an extract of Melia azadirachta (Melia azedarach, Melia tosendan).
The term "pharmaceutically acceptable" as used herein means a physiologically acceptable nontoxic composition which does not inhibit the action of the active ingredient and does not usually cause allergic reactions such as gastrointestinal disorders and vertigo or reactions similar thereto after administration to humans.
The pharmaceutically acceptable salts are preferably pharmaceutically acceptable acid addition salts formed with free acids (free acids). The free acid can be organic acid and inorganic acid. In addition to the organic acids, 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, in addition, hydrochloric acid, bromic 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 assay for Toosendanin (Toosendanin)
In HUVECs, Toosendanin cytotoxicity was examined. HUVECs cell line 1X 104Cells/well were seeded in 96-well plate cell culture plates at 37 ℃ with 5% CO2Incubate for 12-16h in an incubator, treat 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 of Toosendanin, three duplicate wells per group, and add the same amount of dimethyl sulfoxide (DMSO) to the control group. Adding 10 μ l CCK8 after 24h drug action, incubating at 37 deg.C for 1h, and detecting OD450nmAnalyzing the activity of the cells; the results of the detection are shown in FIG. 1.
Wherein CCK8 is a kit for cell activity assay, which is contained in a solution
Is 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, the CC of Toosendanin50Greater than 450. mu.M. In the examples that follow, Toosendanin is used at a concentration of 1-40. mu.M, within the safe and non-toxic range.
Experimental example 2
Detection of antiviral Activity of Toosendanin on SFTSV
Vero cells were grown at 2X 105Cells/well were seeded in 24-well plates at 37 ℃ with 5% CO2Incubate for 12-16h in an incubator, treat with Toosendanin at 5. mu.M, 10. mu.M, 20. mu.M, 40. mu.M, and add the same volume of DMSO as the negative control. After incubation for 1h, SFTSV is used for infection, after incubation for 2h at 37 ℃, a DMEM medium with a culture medium of 2% FBS is replaced, and after 24h, the effect of Toosendanin on inhibiting virus replication is detected through titer, and the result is shown in figure 2, and the compound Toosendanin has a remarkable inhibiting effect on SFTSV replication under different concentration conditions. IC for further inhibiting SFTSV on Toosendanin in Vero cells50The detection is carried out, and the inhibition curve of the SFTSV is shown in figure 3; the round dots in FIG. 3 indicate the inhibition rate of the drug against viral replication, and the triangles indicate cytotoxicity As can be seen from FIG. 3, Toosendanin significantly inhibits the activity of SFTSV, and the IC is calculated50At 1.2. mu.M. CC in Vero cells50Greater than 250 μ M, and SI index greater than 208.3.
The toosendanin has the effect of inhibiting the activity of SFTSV and can be used for preparing the medicine for resisting fever with thrombocytopenia syndrome virus.
Experimental example 3
Effect of Toosendanin on RVFV replication
HUVECs cells were 2X 105Cells/well were seeded in 24-well plates at 37 ℃ with 5% CO2Incubate for 12-16h in an incubator, treat with Toosendanin 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 add the same volume of DMSO as the negative control. After 1h of incubation, infecting with Rift Valley Fever Virus (RVFV) belonging to phlebovirus, after incubating for 2h at 37 ℃, changing DMEM medium with 2% FBS, after 24h, detecting the copy number of the viral gene in the supernatant by a real-time fluorescent quantitative PCR (qPCR) method, as shown in FIG. 4; as can be seen from FIG. 4, the compound Toosendanin has significant inhibitory effect on Rift Valley Fever Virus (RVFV) replication under different concentration conditions; it is indicated that toosendanin has the function of inhibiting the replication of rift valley fever virus.
Experimental example 4
Effect of Toosendanin on SFTSV infection in C57BL/6 mice
C57BL/6, the immune system of the mouse is sound, the genetic background is clean, and the method can be used for drug protective evaluation experiments. The mice were all bred in an SPF environment and were divided into 2 groups: (1) SFTSV + vehicle group (n-5), (2) SFTSV + tosendinin group (n-5). The mouse infection is performed by intraperitoneal injection, and the inoculation dose is 100 mu L (10)5FFU/tube), mock group was an equal volume of DMEM medium. The administration is carried out by intraperitoneal injection, the dosage of the Toosendanin is 1mg/kg/d, and the administration is started 3 days before infection, 1 time per day, and the total time is 6 days. The overall experimental procedure followed strictly the guidelines of the national institutes of health of the united states as set forth in the guidelines for the operation of the animal care and use committee. The mouse daily monitoring indexes include: arch, back, standing hair, motility, stress, weight, etc. Mice were dissected on day 3 of SFTSV infection and spleens were removed to detect viral load. As a result, as shown in FIG. 5, the spleen viral titer of the mice in the administered group was significantly decreased as compared with that of the control group. It was suggested that Toosendanin could significantly inhibit replication of SFTSV at the mouse level.
TSN in FIG. 5 represents SFTSV + Toosendanin group.
Experimental example 5
Effect of Toosendanin on SARS-CoV-2 replication
IC of Toosendanin in Vero-E6 cell for inhibiting SARS-CoV-250And CC50The results of the detection are shown in the SARS-CoV-2 inhibition curve in FIG. 6, the block in FIG. 6 shows the inhibition rate of the drug for inhibiting virus replication, the dot shows cytotoxicity, Toosendanin significantly inhibits the activity of SARS-CoV-2, and the IC is calculated50It was 0.24. mu.M. CC in Vero-E6 cells50Greater than 450 μ M, and an SI index greater than 1875.
It is proved that toosendanin can inhibit the replication of SARS-CoV-2 virus, and can be used for preparing medicine for resisting SARS-CoV-2 virus.
In experimental examples 1 to 5, the present application provides a drug containing toosendanin or a pharmaceutically acceptable salt, solvate or hydrate thereof.
The medicine can be used for resisting bunyavirus, fever with thrombocytopenia syndrome virus, anti-Valley fever virus and SARS-CoV-2 virus.
In conclusion, toosendanin, or a pharmaceutically acceptable salt, solvate or hydrate thereof, is effective against bunyavirus, fever with thrombocytopenia syndrome virus, anti-rift Valley fever virus and anti-SARS-CoV-2 virus.
The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims (4)
1. Use of a composition comprising toosendanin or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of SARS-CoV-2 virus.
2. The use of claim 1, the use of the composition in the preparation of a medicament for inhibiting SARS-CoV-2 virus replication.
3. The use according to claim 1 or 2, the composition further comprising a pharmaceutically acceptable excipient.
4. The use according to claim 1 or 2, wherein the composition is in the form of a gastrointestinal administration form or an 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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| CN202210103516.3A CN114288306B (en) | 2021-07-20 | 2021-07-20 | Use of composition in preparing medicine for inhibiting SARS-CoV-2 virus replication |
| CN202110821896.XA 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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| 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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| CN202210103516.3A Active 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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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100317637A1 (en) * | 2009-06-16 | 2010-12-16 | Industrial Technology Research Institute | Anti-hepatitis c composition and method for preparing drug for inhibiting hepatitis c viruses or treating hepatitis c |
| 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 |
| 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 (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100317637A1 (en) * | 2009-06-16 | 2010-12-16 | Industrial Technology Research Institute | Anti-hepatitis c composition and method for preparing drug for inhibiting hepatitis c viruses or treating hepatitis c |
| 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 (2)
| Title |
|---|
| TAKAKO WATANABE；等: "Inhibitory effect of a triterpenoid compound, with or without alpha interferon, on hepatitis C virus infection" * |
| YOUNG-HEE JIN ；等: "Toosendanin From Melia Fructus Suppresses Influenza A Virus Infection by Altering Nuclear Localization of Viral Polymerase PA Protein" * |
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| CN113350361A (en) | 2021-09-07 |
| CN114288306B (en) | 2023-09-01 |
| CN113350361B (en) | 2022-03-08 |
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