CN111568900A

CN111568900A - Application of indomethacin in resisting coronavirus infection

Info

Publication number: CN111568900A
Application number: CN202010106598.8A
Authority: CN
Inventors: 徐天宏; 桑托罗
Original assignee: Individual
Current assignee: Individual
Priority date: 2020-02-21
Filing date: 2020-02-21
Publication date: 2020-08-25

Abstract

The invention relates to an application of indomethacin in resisting coronavirus infection, and provides a new application of indomethacin in inhibiting SARS coronavirus and a new dosage form thereof, wherein the new dosage form is an inhalation preparation. Indomethacin can remarkably inhibit SARS coronavirus, and is expected to be a drug for treating diseases caused by SARS coronavirus.

Description

Application of indomethacin in resisting coronavirus infection

Technical Field

The invention belongs to the field of drug research and development, and particularly relates to application of indomethacin in resisting coronavirus infection.

Background

Coronaviruses (Coronavirus) are a class of pathogenic single-stranded RNA viruses that infect humans and other vertebrates, causing inflammation of the respiratory and digestive tracts. Coronaviruses belong to the phylogenetic group of the order of the nested viruses (Nidovirales) and the family of Coronaviridae (Coronaviridae) and belong to the genus Coronavirus (Coronavirus), a large group of viruses widely existing in nature. 2019 novel coronavirus (SARS-CoV-2) is currently known as the 7 th coronavirus that can infect humans, and the SARS-CoV coronavirus that has exploded in 2003 is genetically very close to belonging to the same virus species (Table 1) as determined by the International Committee for Classification of viruses, and the remaining 5 coronavirus that infect humans are HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, and MERS-CoV, respectively (causing respiratory syndrome in the middle east).

TABLE 1

The existing treatment method aiming at the lack of effective treatment methods of SARS coronavirus and MERS-CoV is mainly symptomatic treatment, and the existing clinical treatment methods are mainly divided into three types: 1. immune modulatory factors, such as interferon; 2. preventing virus from invading cell molecules, such as monoclonal antibody of S protein, polypeptide aiming at key fusion region of S protein, etc.; 3. and (3) molecules for inhibiting virus replication, such as virus polymerase specific inhibitors, broad-spectrum protease inhibitors and the like. Among these methods, both immunomodulatory factors and broad-spectrum protease inhibitors are non-specific therapies, with large side effects, the sequelae that are easily left in the cured patient, and large individual differences in therapeutic effects (Al-Hazmi, 2016; Zumla et Al, 2016).

The SARS virus has strong survival ability and strong infectivity in the outside world. According to the research results, SARS virus can keep activity for a long time and can be spread in the form of aerosol in the human body common excrement sputum, excrement and blood, SARS-CoV is developed in 2003, and SARS-CoV-2 is developed in large scale in 2019, which brings serious harm to the life, production and life of people, but the therapeutic drug is not disclosed yet. Therefore, it is a great matter to find effective remedy for SARS. However, the development period of brand-new drugs is long, the cost is huge, and the drugs capable of inhibiting the coronavirus are screened from the existing approved drugs (old drugs) until the outbreak of epidemic situation cannot be dealt with.

Indomethacin, also called as indomethacin, has a structural formula shown in figure 1, is a classic non-corticosteroid anti-inflammatory antipyretic and analgesic drug, reduces Prostaglandin (PG) synthesis by inhibiting Cyclooxygenase (COX), inhibits the formation of pain nerve impulses of inflammatory tissues, inhibits inflammatory reactions including the inhibition of leukocyte chemotaxis and the release of lysosomal enzymes, and the like, thereby having antipyretic, analgesic and anti-inflammatory effects. Through search, no report on the anti-coronavirus or SARS-CoV-2 activity of indomethacin is found.

Disclosure of Invention

In order to solve the problem of poor curative effect of the drugs for treating SARS coronavirus infection in the prior art, the invention provides application of indomethacin in SARS coronavirus infection, and provides a new formulation and an inhalation formulation of indomethacin, which are used for treating pneumonia and complications thereof caused by SARS coronavirus.

Indomethacin is also called as indomethacin, and is a clinically common non-corticosteroid anti-inflammatory antipyretic and analgesic.

AI artificial intelligence technology is used to screen the inhibition of SARS-CoV-2 by thousands of old drugs through simulating the signal path of virus and host cell, and indomethacin is found to have strong potential virus inhibition. Further cell level experiments and animal experiments prove that the indomethacin can obviously inhibit the replication of the coronavirus, has an action mechanism different from 3 types of methods aiming at SARS treatment in clinic, is a brand new action mechanism, and can quickly activate protein kinase R in cells so as to activate a cell level virus defense mechanism.

The first aspect of the invention provides an application of indomethacin shown in a structural formula (A) in a medicine for preventing or treating coronavirus, wherein the coronavirus is preferably coronavirus infecting human; further, the coronavirus is preferably SARS-CoV-2, SARS-CoV or a virus of the same species, MERS-CoV

In a second aspect of the invention, there is provided a pharmaceutical composition for inhibiting coronavirus comprising indomethacin and one or more pharmaceutically acceptable excipients, fillers, binders, lubricants, disintegrants, or stabilizers.

In a third aspect of the present invention, an indomethacin inhalant is provided, which is used to increase the effective concentration of indomethacin in local lung cells, has higher bioavailability, avoids the first pass effect of liver, and more effectively treats SARS pneumonia.

The compounds of the present invention or pharmaceutical compositions containing them may be administered in unit dosage form by enteral or parenteral routes, such as by inhalation, oral, intravenous, intramuscular, subcutaneous, nasal, oromucosal, ocular, pulmonary and respiratory, dermal, vaginal, rectal, and the like. The dosage form for administration may be a liquid dosage form, a solid dosage form, or a semi-solid dosage form. The liquid dosage forms can be solution (including true solution and colloidal solution), emulsion (including o/w type, w/o type and multiple emulsion), suspension, injection (including water injection, powder injection and infusion), eye drop, nose drop, lotion, liniment, etc.; the solid dosage form can be tablet (including common tablet, enteric coated tablet, buccal tablet, dispersible tablet, chewable tablet, effervescent tablet, orally disintegrating tablet), capsule (including hard capsule, soft capsule, and enteric coated capsule), granule, powder, pellet, dripping pill, suppository, pellicle, patch, aerosol (powder), spray, etc.; semisolid dosage forms can be ointments, gels, pastes, and the like.

The dosage of the pharmaceutical composition of the compound of the present invention to be administered may vary widely depending on the nature and severity of the disease to be prevented or treated, the individual condition of the patient or animal, the route and dosage form of administration, and the like. Generally, a suitable daily dosage range for a compound of the invention is from 0.001 to 150mg/kg body weight, preferably from 0.1 to 100mg/kg body weight, more preferably from 0.5 to 60mg/kg body weight, most preferably from 1 to 30mg/kg body weight. The above-mentioned dosage may be administered in one dosage unit or divided into several dosage units, depending on the clinical experience of the physician and the dosage regimen including the use of other therapeutic means

Indomethacin has been used clinically for a long time as a classic non-cortical hormone anti-inflammatory antipyretic and analgesic, and has good safety, and the drug metabolism characteristic and the toxic and side effect are very clear. The new application of the medicine for resisting coronavirus can be quickly applied to pathogenic infection of coronavirus, and is particularly used for relieving SARS-CoV-2 serious epidemic outbreak in 2020.

Drawings

FIG. 1: molecular structural formula of indometacin

FIG. 2: in example 1, indomethacin blocked SARS-CoV infection Activity

Detailed Description

EXAMPLE 1 pharmacodynamic test method for anti-SARS-CoV infection

In the invention, SARS-CoV (TOR-2) is used to infect human alveolar epithelial cell line A549. Cell monolayers were plated in 12-well plates and virus yields were determined by PFU and TCID50, and confluent A549 cells were infected with SARS-CoV1 hours at 37 ℃ with a multiplicity of infection (MOI) of 10 PFU/cell. After 1 hour of viral infection, the virus inoculum was removed, washed 3 times with PBS, and different concentrations of indomethacin dissolved in DMSO were added and the cell viability was measured by MTT. The results show that the indometacin can effectively inhibit the infection of SARS-CoV virus to host cells, reduce the generation of virus particles, and has the inhibiting effect in positive correlation with the drug concentration, the half inhibiting concentration (IC50) of the indometacin is 5 mu M (figure 2), and the indometacin can remarkably inhibit SARS-CoV at the final concentration of 400 mu M, and the virus quantity is reduced by nearly 4 Logs. The non-corticoid anti-inflammatory drug aspirin which is the same as indomethacin in inhibiting Cyclooxygenase (COX) does not inhibit coronavirus, thus indicating that the mechanism of inhibiting SARS-CoV by indomethacin is independent of Cyclooxygenase (COX). The above tests relating to SARS-CoV virus with infectivity are all completed in a WHO-compliant P3 level biological safety protection laboratory.

EXAMPLE 2 pharmacodynamic test method for anti-SARS-CoV 2 infection

We can obtain SARS-CoV2 recombinant virus SARS-CoV 2S/HIV with S Protein as shell-wrapped HIV core by co-transfecting SARS-CoV2 envelope S gene (SARS-CoV 2S, GenBank: MN908947.3, plasmid presented by the national institute of biomedical sciences, university of thanksgiving transportation, university of Thailand China) and HIV core plasmid (pNL4-3-Luc-R-E-), respectively, and the construction method is similar to the construction method of SARS-CoV pseudovirus [ Ying Guo, etc. identification of New Region of SARS-CoV S Protein Critical for Viral entry. journal of Molecular Biology 2009,394:600- ]. The pseudovirion is safe because the env, nef and vpr genes on the HIV vector are deleted, so that the virus can only enter host cells but cannot replicate, and the vector is provided with a luciferase reporter gene, so that the degree of infection of the cells by the virus can be indicated by detecting the luciferase activity. After 48 hours of cell infection, the supernatant was discarded, then 100. mu.l of cell lysate (Promega) was added to each well of infected cells to lyse the cells, 50. mu.l of luciferase substrate (Promega) was mixed with 30. mu.l of cell lysate and the relative activity of luciferase was measured using a fluorescence detector, and the intensity of the activity reflected the level of infection by the virus.

The results show that indometacin can effectively inhibit the infection of SARS-CoV-2 virus to host cells, reduce the generation of virus particles, and the inhibition effect is positively correlated with the drug concentration, and the half inhibition concentration (IC50) is 1 mu M (Table 2). Indometacin was not cytotoxic at a concentration of 500. mu.M.

TABLE 2

EXAMPLE 3 Indometacin inhalant form

Indometacin is used as a common non-cortical hormone type anti-inflammatory antipyretic and analgesic, and the main dosage forms at present are various oral dosage forms, external liniments, suppositories and the like, and no inhalation dosage forms are available. The invention provides an indomethacin inhalant for increasing the effective concentration of indomethacin in local lung cells, having higher bioavailability, avoiding the first pass effect of liver and more effectively treating pneumonia caused by factors such as SARS virus infection and the like.

The water solubility of the indomethacin is poor, sodium carbonate is added into normal saline, the pH value of the solution is adjusted to 7.4, the indomethacin is slowly added to form indomethacin sodium salt, the water solubility of the indomethacin sodium salt can be obviously enhanced, the indomethacin sodium salt is prepared into an indomethacin atomized inhalation solution with the concentration of 20mg/ML, 2ML of the indomethacin atomized inhalation solution is taken out and is administrated by a driving sprayer, and the indomethacin atomized inhalation solution is evaluated by a pharmacopoeia twinimpinger method. The results show that the aerosol particles formed by the method have the diameter of 1.0uM-6.0uM and the deposition rate of the aerosol particles in the parts corresponding to the oral cavity and the throat is 1.2 percent; the deposition rate in the area corresponding to the trachea and the lung is 39.5%.

While the invention has been described with respect to a preferred embodiment, it will be understood by those skilled in the art that the foregoing and other changes, omissions and deviations in the form and detail thereof may be made without departing from the scope of this invention. Those skilled in the art can make various changes, modifications and equivalent arrangements, which are equivalent to the embodiments of the present invention, without departing from the spirit and scope of the present invention, and which may be made by utilizing the techniques disclosed above; meanwhile, any changes, modifications and variations of the above-described embodiments, which are equivalent to those of the technical spirit of the present invention, are within the scope of the technical solution of the present invention.

Claims

1. Application of indomethacin in preparing medicine for preventing and treating coronavirus infection is provided.

2. The application of a pharmaceutical composition in preparing a medicament for preventing or treating coronavirus infection is characterized in that the pharmaceutical composition contains indometacin and a pharmaceutically acceptable carrier or excipient.

3. An indomethacin inhalant and its application in treating respiratory tract inflammation caused by coronavirus are provided.

4. The use according to claim 2, wherein said pharmaceutical composition further comprises an additional antiviral agent.

5. The product and use according to any one of claims 1 to 3, characterized in that said coronavirus is a human-infecting coronavirus.

6. The product and use according to any of claims 1-3, characterized in that the coronavirus is selected from the group consisting of SARS-CoV, SARS-CoV2, MERS-CoV or a common cold causing coronavirus.

7. Use according to any one of claims 1 to 3, wherein said coronavirus is selected from the group consisting of SARS-CoV, SARS-CoV2 or a virus belonging to the same species (speces) as it belongs to the International Commission of viral Classification (ICTV) coronavirus classification.