CN111420024A - Application of bacitracin A in preparing medicine for preventing and treating coronavirus - Google Patents
Application of bacitracin A in preparing medicine for preventing and treating coronavirus Download PDFInfo
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- CN111420024A CN111420024A CN202010265260.7A CN202010265260A CN111420024A CN 111420024 A CN111420024 A CN 111420024A CN 202010265260 A CN202010265260 A CN 202010265260A CN 111420024 A CN111420024 A CN 111420024A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/12—Cyclic peptides, e.g. bacitracins; Polymyxins; Gramicidins S, C; Tyrocidins A, B or C
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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
- A61P11/00—Drugs for disorders of the respiratory system
- A61P11/14—Antitussive agents
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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
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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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 relates to application of bacitracin A in preparing medicaments for preventing and treating coronavirus, and particularly discloses application of bacitracin A or pharmaceutically acceptable salts, isotopes, stereoisomers, mixtures of stereoisomers, tautomers, esters, amides or prodrugs thereof in preparing medicaments 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, the half effective concentration (EC50) of bacitracin A on novel coronavirus (SARS-Cov-2) is 2.38 mu M, and the toxicity is low, which indicates that the bacitracin A has a good treatment window.
Description
Technical Field
The invention belongs to the field of antiviral drugs, and particularly relates to application of bacitracin A in preparation of a drug for preventing and treating 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 development of effective antiviral drugs and vaccines is the most urgent task at present, the treatment of COVID-19 mainly depends on the adjuvant therapy for the symptoms, but lacks effective specific drugs and vaccines, however, the development of new drugs and vaccine is a time-consuming process, not only has a long development period, but also is a time-consuming project after the approval of the market, the search for "old drugs" having the effect of resisting infection of SARS-Cov-2 virus from the marketed drugs is an effective strategy for treating and preventing the outbreak disease of the COVID-2 virus, the main anti-SARS-Cov-2 virus includes RNA inhibitor, polymerase inhibitor, β interferon, monoclonal antibody, etc., and the analysis of the activity of these drugs and vaccines in vitro is not yet effective50In vivo efficacy is still being observed between low micromolar to medium micromolar.
Baceride a is a cyclic polypeptide produced by Bacillus subtilis var organisms that inhibits the proliferation of gram-positive bacteria by interfering with the synthesis of cell wall peptidoglycan. Phthalide a affects bacterial biochemical processes including metal ion migration, peptidoglycan synthesis, cell membrane permeability and synthesis of inducible enzymes. In addition to antibacterial activity, bacitracin a has been demonstrated as a tool for studying lipid-protein interactions at the molecular level.
The development of effective specific drugs for treating coronavirus pneumonia is an urgent problem to be solved. In the aspect of developing a medicine aiming at a novel coronavirus (SARS-Cov-2) virus, the invention researches the effect of the bacitracin A on resisting SARS-Cov-2 virus.
Disclosure of Invention
The invention aims to provide application of the bacitracin A in preparing a medicament for preventing or treating diseases caused by coronavirus.
Specifically, to solve the technical problem of the present invention, the following technical solutions are adopted:
the invention provides application of bacitracin A or pharmaceutically acceptable salt, isotope, stereoisomer, mixture of stereoisomer, tautomer, ester, amide or prodrug thereof in preparing medicaments 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, OC43, HKU1 or MERS-CoV, and is preferably respiratory tract infectious disease, such as severe acute respiratory syndrome, severe acute respiratory syndrome coronavirus type 2 and respiratory syndrome of middle east.
In the technical scheme of the invention, the bacitracin A is shown as a structural formula (1)
In the technical scheme of the invention, the bacitracin A or pharmaceutically acceptable salt, isotope, stereoisomer, mixture of stereoisomers, tautomer, ester, amide or prodrug thereof is used as the only active ingredient in the preparation of the drugs for preventing and/or treating diseases caused by coronavirus.
In the technical scheme of the invention, the bacitracin A or the pharmaceutically acceptable salt, isotope, stereoisomer, mixture of stereoisomers, tautomer, ester, amide or prodrug thereof and other antiviral drugs are used for preparing the composition which is used as an active ingredient in the preparation of the drugs for preventing and/or treating the diseases caused by the coronavirus.
In the technical scheme of the invention, the other antiviral drugs are selected from ganciclovir, acyclovir, amantadine, oseltamivir, abacavir, acemannan, acyclovir sodium, adefovir, alovudine, avsunotol, tricyclodecylamine hydrochloride, alaudine, alitame, adefovir, cidofenac, emtricitabine, cytarabine hydrochloride, delavirdine mesylate, desciclovir, didanosine, dioxaxaline, edexuridine, virin, itracetipine, envilarden, engivir oxime, hopudine, famciclovir hydrochloride, clofenacil, fossilidine, foscarnet sodium, ganciclovir sodium, idoxuridine, indinavir, ethoxybutotid, lamivudine, lomicrivir, locarine, lovir adenosine, lopinavir, Memantine hydrochloride, methylthioninium chloride, nelfinavir, nevirapine, penciclovir, pirodapvir, 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.
In another aspect of the present invention, there is provided a pharmaceutical composition for treating or preventing diseases caused by viruses of the family Coronaviridae, comprising bacerianin A 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.
In another aspect of the invention, there is provided the use of bacitracin a 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 bacerian A.
Another aspect of the 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 bacerianine a or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture of stereoisomers, tautomer, ester, amide or prodrug thereof.
Another aspect of the 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 bacerianin a, or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture of stereoisomers, tautomer, ester or prodrug thereof, prior to infection.
The invention discloses a medicine using bacillphthalein A as main component and its application in resisting new coronary pneumonia (COVID-19) new type coronavirus (SARS-Cov-2) infection. The invention discovers for the first time that the bacitracin A has antiviral effect on the novel coronavirus, can block the novel coronavirus from infecting host cells, and can be used for treating diseases for resisting the novel coronavirus infection of the novel coronavirus.
Advantageous effects
The bacerianin A has antiviral effect on new coronavirus, can block host cell infection caused by new coronavirus, and can be used for treating diseases caused by new coronavirus infection. Half Effective Concentration (EC) of bacitracin A against novel coronavirus of New crown pneumonia (SARS-Cov-2)50) 2.38 mu M, low toxicity and a good therapeutic 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
Will be notVeroE6 cells were 3 × 10% w/w5One well, inoculated into a 96-well plate, placed in minimal Eagle's minimal basal medium (MEM; GibcoInvitrogen) containing 10% fetal bovine serum (FBS; GibcoInvitrogen) at 37 ℃ with 5% CO2Culturing until the monolayer grows. Diluting new coronavirus clinical isolate 100 times, inoculating to 96-well plate full of monolayer 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 the toxicity of bacitracin A drugs
Dissolving bacilphthalein A powder with DMSO, adding culture solution to dilute 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 medicaments with different dilutions in a ratio of l00 mu L per well, adding MEM in a ratio of l00 mu L per well in 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 00 mu L dimethyl sulfoxide (DMSO) into each hole, oscillating at a 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 efficacy of phthalide A against the novel coronavirus of New Copyneumoniae
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 dilutions of drug were added, 4 multiple wells per concentration, at 200 μM is the initial concentration of the drug at 34 ℃ with 5% CO2Incubate in incubator 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) Half Effective Concentration (EC) for the novel coronavirus of New coronary pneumonia (SARS-Cov-2)50) 2.38 mu M, and low half effective concentration, which indicates that the antiviral effect is higher.
Example 4 immunofluorescence assay
Immunofluorescence microscopy to detect the 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 with secondary antibody incubation antibody (488Affinipure Donkey Anti-RabbitIgG (H + L), cell nuclei were stained with Hoechst33258 dye (Beyotime, China). Bacillary phthalein A 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 the bacerianin A or its pharmaceutically acceptable salt, isotope, stereoisomer, mixture of stereoisomers, tautomer, ester, amide or prodrug in preparing medicine 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. Use according to claim 1 of bacerianin a or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture of stereoisomers, tautomer, ester, amide or prodrug thereof as sole active ingredient for the preparation of a medicament for the prophylaxis and/or treatment of diseases caused by coronaviruses; or
The application of the bacerianin A or the pharmaceutically acceptable salt, isotope, stereoisomer, mixture of stereoisomer, tautomer, ester, amide or prodrug thereof and the composition of other antiviral drugs as active ingredients 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, alitame, adefovir, cidofophylline, emtricitabine, cytarabine hydrochloride, delavirdine mesylate, desciclovir, didanosine, dioxazoline, edexuridine, emivirin, itracitalopine, emviraden, envirox, greet, famciclovir, cloquine hydrochloride, fosetyl, fosamivir, fosalvudine sodium, ganciclovir sodium, indinavir, ethoxybutovidone, lamivudine, lombara, lodenadine, lomavine hydrochloride, mavir hydrochloride, mavir, mafenib, mavir, ma, Nevirapine, penciclovir, pirodavir, ribavirin, saquinavir mesylate, ritonavir, hydrochloric acid of sothiamide, solivudine, stolonicin, stavudine, tenofovir, hydrochloric acid of trovudine, valacyclovir hydrochloride, vidarabine, adenosine phosphate, vidarabine sodium phosphate, tipranavir, viruoxime, zalcitabine, zidovudine, net oxime.
5. A pharmaceutical composition for the treatment or prevention of a disease caused by a coronavirus, comprising bacerianine a 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 of viruses of the family Coronaviridae, said disinfectant comprising bacerianin A.
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 bacerianine a 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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| CN202010265260.7A CN111420024B (en) | 2020-04-07 | 2020-04-07 | Application of bacillus phthalein A in preparing medicament for preventing and treating coronavirus |
| PCT/CN2020/129186 WO2021203704A1 (en) | 2020-04-07 | 2020-11-16 | Use of bacitracin a in preparation of drugs for preventing and treating coronaviruses |
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| CN202010265260.7A CN111420024B (en) | 2020-04-07 | 2020-04-07 | Application of bacillus phthalein A in preparing medicament for preventing and treating coronavirus |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113350482A (en) * | 2021-05-31 | 2021-09-07 | 中国食品药品检定研究院 | Application of phytohemagglutinin PHA-E in preparation of medicine for treating diseases caused by coronavirus |
| WO2021203704A1 (en) * | 2020-04-07 | 2021-10-14 | 中国科学院深圳先进技术研究院 | Use of bacitracin a in preparation of drugs for preventing and treating coronaviruses |
| 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 |
| 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 |
| WO2022118020A1 (en) * | 2020-12-02 | 2022-06-09 | Cipla Limited | Method of treating viral infection |
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| WO2021203704A1 (en) * | 2020-04-07 | 2021-10-14 | 中国科学院深圳先进技术研究院 | Use of bacitracin a in preparation of drugs for preventing and treating coronaviruses |
| 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 |
| 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 |
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| CN113350482A (en) * | 2021-05-31 | 2021-09-07 | 中国食品药品检定研究院 | Application of phytohemagglutinin PHA-E in preparation of medicine for treating diseases caused by coronavirus |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2021203704A1 (en) | 2021-10-14 |
| WO2021203704A9 (en) | 2021-11-25 |
| CN111420024B (en) | 2023-10-03 |
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