CN111467363A - Application of sofosbuvir in preparation of medicine for preventing and treating coronavirus - Google Patents
Application of sofosbuvir in preparation of medicine for preventing and treating coronavirus Download PDFInfo
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- CN111467363A CN111467363A CN202010266185.6A CN202010266185A CN111467363A CN 111467363 A CN111467363 A CN 111467363A CN 202010266185 A CN202010266185 A CN 202010266185A CN 111467363 A CN111467363 A CN 111467363A
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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
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
- A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
- A61K31/7064—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines
- A61K31/7068—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid
- A61K31/7072—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid having two oxo groups directly attached to the pyrimidine ring, e.g. uridine, uridylic acid, thymidine, zidovudine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
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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
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
Abstract
The invention discloses an application of sofosbuvir in preparing a medicament for preventing and treating coronavirus, in particular an application of sofosbuvir or pharmaceutically acceptable salts, isotopes, stereoisomers, a mixture of stereoisomers, tautomers, esters, amides or prodrugs thereof in preparing a medicament for preventing and/or treating diseases caused by coronavirus, wherein the coronavirus is novel coronavirus SARS-Cov-2, SARS-CoV, HCoV229E, N L63, OC43, HKU1 and MERS-CoV.
Description
Technical Field
The invention belongs to the field of antiviral drugs, and particularly relates to application of sofosbuvir in preparation of drugs for preventing and treating anti-coronavirus.
Background
New Coronaviridae (COVID-19) is an infectious disease caused by infecting a human body with a novel coronavirus (SARS-Cov-2), and The symptoms thereof mainly include fever, weakness, dry cough, dyspnea and renal failure [ The L ancet,2020,395(10223): 507-; The L ancet,2020,395(10223):497 506 ]. Coronaviridae (Coronaviridae) belonging to The family Coronaviridae (Coronaviridae) in The systematic classification, Coronaviridae (Corona viruses) belonging to The genus coronaviruses are positive strand single strand RNA viruses with mantle (Envelope), The diameter thereof is about 80-120 nm, The genetic material thereof is The largest of all RNA viruses, and The coronaviruses are generally only infected with human, mouse, pig, cat, dog, vertebrate, coronavirus which was first isolated from chicken in 1937, The glycoprotein particle shape is irregular, The diameter thereof is about 60, 220nm, The glycoprotein has The structure of The above glycoprotein (glycoprotein A-19, glycoprotein A) and glycoprotein (glycoprotein A) which are also included in The glycoprotein types of fowl (glycoprotein).
The gene group sequencing shows that SARS-Cov-2 is a single-chain RNA coronavirus with the diameter of 60-140 nm and 9-12 nm prickle outside the envelope, and is similar to corolla, and the comparison with other virus sample gene sequences shows that SARS-Cov-2 is similar to SARS-CoV (79.5%) [ Nature,2020] and bat coronavirus (96%) [ bioRxiv,2020,2020.01.22.914952 ], and the virus is presumed to possibly originate from bat [ bioRxiv,2020,2020.01.24.915157; Nature,2020 ]. SARS-Cov-2 belongs to β CoV, which is the 7 th member of HCoV family different from SARS-CoV and MERS-CoV [ New England of medicinal, 2020], and the other 6 members include HCoV 36229 54, OC 57, HKU 82, SARS-Cov and MERS-CoV.
The new type of coronavirus causing pneumonia is a new type of coronavirus, which is well known to cause atypical pneumoniaThe development of effective antiviral drugs and vaccines is the most urgent task at present, the treatment of COVID-19 mainly relies on the symptomatic adjuvant therapy, and effective specific drugs and vaccines are not yet lacked, however, the development of new drugs and the preparation of vaccines are a time-consuming process, not only the development cycle is long, but also the mass production after approval on the market is a time-consuming project, the search for "old drugs" having the effect of resisting SARS-CoV-2 virus infection from the marketed drugs is an effective strategy for treating and preventing the explosive infection disease of COVID-19, the anti-SARS-CoV-2 virus drugs currently under study mainly include RNA polymerase inhibitors, β interferons, monoclonal antibodies, vaccines and the like, however, until now, there is no effective antiviral drugs and vaccines50In vivo efficacy is still being observed between low micromolar to medium micromolar.
Sofosbuvir was developed by Gilidard (Gilead), and was marketed by the Food and Drug Administration (FDA) on 6.12.2013, then approved by the European drug administration (EMA) on 16.1.2014, and then approved by the Japanese pharmaceutical and medical device integration agency (PMDA) on 26.3.2015, and marketed by Gilidard under the trade name of Gilidard
Sofosbuvir is an HCV NS5B RNA-dependent RNA polymerase inhibitor, which plays an important role in viral replication. In view of the action mechanism of the medicine, and on the basis of the virtual screening result of the medicine, the application research of sofosbuvir in the antiviral effect of the novel coronavirus of the new coronary pneumonia is carried out.
The development of effective specific drugs for treating coronavirus pneumonia is an urgent problem to be solved. In the aspect of research and development of a medicine aiming at a novel coronavirus (SARS-Cov-2) virus, the invention researches the effect of sofosbuvir on resisting SARS-Cov-2 virus.
Disclosure of Invention
The invention aims to provide application of sofosbuvir in preparation of a medicament for preventing or treating diseases caused by coronavirus.
Specifically, in order to solve the technical problem of the present invention, the following technical scheme is adopted:
the invention provides an application of sofosbuvir or pharmaceutically acceptable salts, isotopes, stereoisomers, mixtures of stereoisomers, tautomers, esters, amides or prodrugs thereof in preparing a medicament for preventing and/or treating diseases caused by coronavirus.
In the technical scheme of the invention, the coronavirus is novel coronavirus SARS-Cov-2, SARS-CoV, HCoV229E, N L63, OC43, HKU1 and MERS-CoV.
In the technical scheme of the invention, the disease caused by the coronavirus is infectious disease or complication thereof caused by any virus of SARS-Cov-2, SARS-CoV, HCoV229E, N L63, HCoV-OC43, HKU1 or MERS-CoV, and is preferably respiratory tract infection disease, such as severe acute respiratory syndrome, severe acute respiratory syndrome coronavirus type 2 and middle east respiratory syndrome.
In the technical scheme of the invention, the Sofosbuvir is shown as a structural formula (1)
In the technical scheme of the invention, sofosbuvir or pharmaceutically acceptable salts, isotopes, stereoisomers, mixtures of stereoisomers, tautomers, esters, amides or prodrugs thereof are used as the only active ingredient in the preparation of medicines for preventing and/or treating diseases caused by coronavirus.
In the technical scheme of the invention, the sofosbuvir or pharmaceutically acceptable salts, isotopes, stereoisomers, mixtures of stereoisomers, tautomers, esters, amides or prodrugs thereof and other antiviral drugs are used for preparing the composition as an active ingredient in the preparation of drugs for preventing and/or treating diseases caused by coronavirus.
In the technical scheme of the invention, the other antiviral drugs are selected from other antiviral drugs selected from ganciclovir, acyclovir, amantadine, rimantadine, oseltamivir, abacavir, acemannan, acyclovir sodium, adefovir, alovudine, avsunotol, amantadine hydrochloride, alafenadine, oreridone, atidine mesylate, avridine, cidofovir, simperidone, emtricitabine, cytarabine hydrochloride, delavirdine mesylate, desciclovir, didanosine, dioxaxaline, edexuridine, emivirin, itracetirizine, emviran, emtricitabine, greevudine, famciclovir, clofenacil hydrochloride, filcitabine, fexuridine, fossilite, foscamet sodium, ganciclovir sodium, idoxuridine, indinavir, ethoxybutanedione, lamivudine, bocavir, lodenosine, valprozin, valprovir, and lodenosine, Lopinavir, memantine hydrochloride, methylthiourea, nelfinavir, nevirapine, penciclovir, pirodavir, ribavirin, saquinavir mesylate, ritonavir, sotalomide hydrochloride, solivudine, penicillin, stavudine, tenofovir, trovudine hydrochloride, valacyclovir hydrochloride, vidarabine phosphate, vidarabine sodium phosphate, tipranavir, viroxime, zalcitabine, zidovudine, and neat oxime.
Another aspect of the present invention provides a pharmaceutical composition for treating or preventing diseases caused by viruses of the family coronaviridae, which comprises sofosbuvir or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture of stereoisomers, tautomer, ester, amide or prodrug thereof.
In the technical scheme of the invention, the pharmaceutical composition also comprises pharmaceutically acceptable auxiliary materials.
In the technical scheme of the invention, the dosage form of the pharmaceutical composition is oral preparation, lung inhalation preparation, mucosa administration preparation, eye preparation or injection.
In the technical scheme of the invention, the oral preparation is selected from granules, powder, pills, tablets, capsules or oral liquid.
Another aspect of the invention provides the use of sofosbuvir as a disinfectant against viruses of the family coronaviridae.
In another aspect of the invention, a disinfectant for eliminating contamination by viruses of the family Coronaviridae is provided, the disinfectant comprising sofosbuvir.
Another aspect of the present invention provides a method for treating a disease caused by a virus of the family coronaviridae, comprising administering to a subject a therapeutically effective amount of sofosbuvir or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture of stereoisomers, tautomers, ester, amide or prodrug thereof.
Another aspect of the present invention provides a method for preventing infection of a subject with a virus of the family coronaviridae, comprising administering to the subject a therapeutically effective amount of sofosbuvir or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture of stereoisomers, tautomer, ester or prodrug thereof, prior to infection.
In the technical scheme of the invention, the medicine or the medicine composition does not contain glucocorticoid or mineralocorticoid.
Advantageous effects
The invention discovers for the first time that sofosbuvir has an antiviral effect on novel coronavirus, can block the novel coronavirus from infecting host cells, and can be used for treating diseases in the aspect of resisting coronavirus infection.
Half the Effective Concentration (EC) of sofosbuvir against the novel coronavirus of new coronary pneumonia (SARS-Cov-2)50) 1.12 mu M, low half effective concentration and low toxicity, and has a good treatment window.
Detailed Description
In order to make the aforementioned objects, features and advantages of the present invention more comprehensible, specific embodiments of the present invention are described in detail below, but the present invention is not to be construed as limiting the implementable range of the present invention.
Example 1 viral amplification
VeroE6 Vero cells were transformed into 3 × 105Is/areWells were inoculated into 96-well plates and placed at 37 ℃ in a minimum Eagle' medium (MEM; Gibco Invitrogen) containing 10% fetal bovine serum (FBS; Gibco Invitrogen) with 5% CO2Culturing until the monolayer grows. Diluting new coronavirus clinical isolate 100 times, inoculating to 96-well plate full of single-layer cells, placing at 37 deg.C and 5% CO2Two days of culture (containing normal control).
After two days, the pathological change degree reaches over 75 percent, the mixture is placed in an ultra-low temperature refrigerator at minus 80 ℃, freeze thawing is carried out repeatedly for one time, virus liquid of cell amplification is collected, centrifugation is carried out for 30 minutes at 3000r/min, precipitates are removed, and the mixture is subpackaged into small tubes to be placed in the ultra-low temperature refrigerator at minus 80 ℃ for long-term storage.
Example 2 evaluation of Sofosbuvir drug toxicity
Dissolving Sofosbuvir powder in DMSO, diluting with culture medium to 20mg/m L final concentration of 1%, filtering with 0.22 μm filter membrane, storing at 4 deg.C, and storing at 2.5 × 10 per well4Inoculating the cells to a 96-well plate, removing culture solution after the cells grow into a monolayer after 24-48 h, adding medicines with different dilutions into 100 mu L/well, adding 100 mu L/well MEM into a normal cell control well, and adding 5% CO at 37 DEG C2Continuously culturing for 2-5 days, adding CCK8 method solution (5mg/m L) 20 μ L per well, and placing at 37 deg.C with 5% CO2And (3) continuously incubating for 4 hours in the incubator, removing culture supernatant, adding 100 mu L dimethyl sulfoxide (DMSO) into each hole, oscillating at low speed for 10 minutes to fully dissolve crystals, selecting 490nm wavelength, and determining the light absorption value of each hole on an enzyme-linked immunosorbent assay monitor.
Example 3 evaluation of the drug efficacy of Sofosbuvir against the novel coronavirus
To evaluate the antiviral efficacy of the drugs, VeroE6 cells were grown at a density of 5 × 104Cells/well in 48-well cell culture dishes overnight. The virus (MOI 0.05) was added and allowed to infect for 2 hours. Then 2-fold gradient diluted drug was added, 4 multiple wells were set for each concentration, 200 μ M was used as the initial concentration of drug, 5% CO at 34 deg.C2IncubatorAnd breeding for 2 days. Cytopathic Effect (CPE) was recorded, CPE was recorded in the cells according to 6-grade criteria, after CPE was recorded, staining was performed by CCK8 method, OD was measured, and half effective concentration of drug (EC) was calculated by Reed-Muench method50) At 1.12. mu.M. The experiment result shows that sodium foscarnet has low antiviral half effective concentration and good antiviral effect.
Example 4 immunofluorescence assay
Immunofluorescence microscopy to detect expression of viral proteins in VeroE6 cells, cells were fixed with 4% paraformaldehyde and permeabilized with 0.5% TritonX-100, then cells were blocked with 5% Bovine Serum Albumin (BSA) at room temperature for 2 hours, cells were incubated with primary antibody (viral nucleocapsid protein polyclonal antibody of Bat SARS-associated coronavirus, Anti-NP, at 1: 1000 dilution) for 2 hours, then incubated with secondary antibody (488Affinipure Donkey Anti-RabbitIgG (H + L), nuclei were stained with Hoechst33258 dye (Beyotime, China), Sofosbuvir was found to be effective in killing viruses in cells by fluorescence microscopy and had little effect on cells with a good therapeutic window.
Claims (10)
1. The application of sofosbuvir or a pharmaceutically acceptable salt, isotope, stereoisomer, a mixture of stereoisomers, tautomer, ester, amide or prodrug thereof in preparing a medicament for preventing and/or treating diseases caused by coronavirus.
2. The use of claim 1, wherein the coronavirus is a novel coronavirus, SARS-Cov-2, SARS-CoV, HCoV229E, N L63, OC43, HKU1 and MERS-CoV.
3. The use according to claim 1, wherein the disease caused by coronavirus is infectious disease caused by SARS-Cov-2, SARS-CoV, HCoV229E, N L63, HCoV-OC43, HKU1 or MERS-CoV or its complications, and the infectious disease is preferably respiratory infectious disease.
4. The use according to claim 1 of sofosbuvir or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture of stereoisomers, tautomer, ester, amide or prodrug thereof as sole active ingredient in the manufacture of a medicament for the prophylaxis and/or treatment of diseases caused by coronaviruses; or
The application of the composition of sofosbuvir or pharmaceutically acceptable salts, isotopes, stereoisomers, mixture of stereoisomers, tautomers, esters, amides or prodrugs thereof and other antiviral drugs as an active ingredient in the preparation of drugs for preventing and/or treating diseases caused by coronavirus;
preferably, the other antiviral drug is selected from the group consisting of ganciclovir, acyclovir, amantadine, rimantadine, oseltamivir, abacavir, acemenan, acyclovir sodium, adefovir, alovudine, avrinol, amantadine hydrochloride, amantadine, aliquodine mesylate, avridine, cidofovir, cidofophylline, emtricitabine, cytarabine hydrochloride, delavirdine mesylate, desciclovir, didanosine, dioxazoline, edexuridine, emivirin, itracitabine, emviraden, enviroxime, hoplatin, famciclovir, cloquine hydrochloride, decitabine, fexitabine, fexiuridine, foscarnet sodium, fosamivir sodium, idovir sodium, idoside, indinavir, ethoxybutovidone aldehyde, lamivudine, lobbucavir, lodenafine hydrochloride, lopinavir, mavir hydrochloride, mavir sodium acetate, trexate, trematodine hydrochloride, tretinomycin hydrochloride, and sodium acetate, Nelfinavir, nevirapine, penciclovir, pirodavir, ribavirin, saquinavir mesylate, ritonavir, hydrochloric acid of sodamide, solivudine, bractenocillin, stavudine, tenofovir, hydrochloric acid of trovudine, valacyclovir hydrochloride, vidarabine, adenosine phosphate, vidarabine sodium phosphate, tipranavir, viroxim, zalcitabine, zidovudine, net oxime.
5. A pharmaceutical composition for the treatment or prevention of a disease caused by a coronavirus, comprising sofosbuvir or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture of stereoisomers, tautomer, ester, amide or prodrug thereof.
6. The pharmaceutical composition of claim 5, further comprising a pharmaceutically acceptable excipient.
7. The pharmaceutical composition according to claim 5, wherein the pharmaceutical composition is in the form of an oral preparation, a pulmonary inhalation preparation, a mucosal administration preparation, an ophthalmic preparation or an injection; preferably, the oral preparation is selected from granules, powders, pills, tablets, capsules or oral liquids.
8. A disinfectant for eliminating contamination by viruses of the family coronaviridae, comprising sofosbuvir.
9. A method for treating or preventing a disease caused by a virus of the family coronaviridae, comprising administering to a subject a therapeutically effective amount of sofosbuvir or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture of stereoisomers, tautomer, ester, amide or prodrug thereof.
10. The pharmaceutical composition of any one of claims 5-7 or the disinfectant of claim 8 or the method of claim 9, wherein the coronavirus is a novel coronavirus, SARS-Cov-2, SARS-Cov, HCoV229E, N L63, OC43, HKU1, and MERS-Cov.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
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| CN202010266185.6A CN111467363A (en) | 2020-04-07 | 2020-04-07 | Application of sofosbuvir in preparation of medicine for preventing and treating coronavirus |
| PCT/CN2020/129175 WO2021203701A1 (en) | 2020-04-07 | 2020-11-16 | Application of sofosbuvir in preparation of drug for preventing and treating coronavirus |
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| CN112237584A (en) * | 2020-08-21 | 2021-01-19 | 中国医学科学院医药生物技术研究所 | Novel use of compound for preventing and/or treating diseases caused by coronavirus infection |
| CN112661801A (en) * | 2020-12-18 | 2021-04-16 | 成都阿奇生物医药科技有限公司 | Nucleoside analogue and deuteron thereof, and preparation method and application thereof |
| WO2021155119A3 (en) * | 2020-01-29 | 2021-09-10 | The Trustees Of Columbia University In The City Of New York | Therapeutics for covid-19 |
| WO2021203701A1 (en) * | 2020-04-07 | 2021-10-14 | 中国科学院深圳先进技术研究院 | Application of sofosbuvir in preparation of drug for preventing and treating coronavirus |
| WO2022053164A1 (en) * | 2020-09-14 | 2022-03-17 | Hc Pharma Ag | Antiviral combination for the treatment of covid-19 infection |
| WO2022088038A1 (en) * | 2020-10-30 | 2022-05-05 | 中国科学院深圳先进技术研究院 | Application of cay10603 in preparation of drugs for preventing and treating coronavirus-related diseases |
| WO2022088037A1 (en) * | 2020-10-30 | 2022-05-05 | 中国科学院深圳先进技术研究院 | Application of sirtinol in preparation of drug for preventing and treating coronavirus |
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| CN111467363A (en) * | 2020-04-07 | 2020-07-31 | 中国科学院深圳先进技术研究院 | Application of sofosbuvir in preparation of medicine for preventing and treating coronavirus |
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- 2020-11-16 WO PCT/CN2020/129175 patent/WO2021203701A1/en active Application Filing
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2021155119A3 (en) * | 2020-01-29 | 2021-09-10 | The Trustees Of Columbia University In The City Of New York | Therapeutics for covid-19 |
| WO2021203701A1 (en) * | 2020-04-07 | 2021-10-14 | 中国科学院深圳先进技术研究院 | Application of sofosbuvir in preparation of drug for preventing and treating coronavirus |
| CN112237584A (en) * | 2020-08-21 | 2021-01-19 | 中国医学科学院医药生物技术研究所 | Novel use of compound for preventing and/or treating diseases caused by coronavirus infection |
| CN112237584B (en) * | 2020-08-21 | 2021-10-01 | 中国医学科学院医药生物技术研究所 | Use of compounds for the prophylaxis and/or treatment of diseases caused by coronavirus infection |
| WO2022053164A1 (en) * | 2020-09-14 | 2022-03-17 | Hc Pharma Ag | Antiviral combination for the treatment of covid-19 infection |
| WO2022088038A1 (en) * | 2020-10-30 | 2022-05-05 | 中国科学院深圳先进技术研究院 | Application of cay10603 in preparation of drugs for preventing and treating coronavirus-related diseases |
| WO2022088037A1 (en) * | 2020-10-30 | 2022-05-05 | 中国科学院深圳先进技术研究院 | Application of sirtinol in preparation of drug for preventing and treating coronavirus |
| CN112661801A (en) * | 2020-12-18 | 2021-04-16 | 成都阿奇生物医药科技有限公司 | Nucleoside analogue and deuteron thereof, and preparation method and application thereof |
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|---|---|
| WO2021203701A1 (en) | 2021-10-14 |
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Application publication date: 20200731 |