阅读说明：本技术 用于预防或治疗严重急性呼吸综合征冠状病毒2感染疾病的组合物 (Composition for preventing or treating severe acute respiratory syndrome coronavirus 2 infection disease ) 是由 郑钟善 洪宗喜 金东明 朴奉焕 柳永培 权亨俊 李仁哲 朴智英 于 2020-10-20 设计创作，主要内容包括：本发明提供用于预防或治疗严重急性呼吸综合征冠状病毒2感染疾病的药学组合物,其包含由化学式1表示的化合物或其药学上可接受的盐作为有效成分。(The present invention provides a pharmaceutical composition for preventing or treating severe acute respiratory syndrome coronavirus 2 infection diseases, comprising a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof as an effective ingredient.)

1. A pharmaceutical composition for preventing or treating severe acute respiratory syndrome coronavirus 2 infectious diseases, characterized by,

comprises the following steps:

a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof; and

a compound represented by the following chemical formula 2:

chemical formula 1:

chemical formula 2:

2. the pharmaceutical composition for preventing or treating severe acute respiratory syndrome coronavirus 2 infectious disease according to claim 1, wherein the compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof and the compound represented by the above chemical formula 2 or a pharmaceutically acceptable salt thereof are administered in combination simultaneously, separately or sequentially.

3. The pharmaceutical composition for preventing or treating Severe acute respiratory syndrome coronavirus 2 infectious disease according to claim 1, wherein Severe acute respiratory syndrome coronavirus 2 infectious disease is a respiratory disease.

4. The pharmaceutical composition for preventing or treating Severe acute respiratory syndrome coronavirus 2 infectious disease according to claim 1, wherein Severe acute respiratory syndrome coronavirus 2 infectious disease shows symptoms after a latency period of 2 to 14 days after infection with said coronavirus.

5. The pharmaceutical composition for preventing or treating Severe acute respiratory syndrome coronavirus 2 infectious disease according to claim 1, wherein Severe acute respiratory syndrome coronavirus 2 infectious disease is a disease infected with Severe acute respiratory syndrome coronavirus 2, and shows one or more symptoms selected from the group consisting of hyperpyrexia, cough, shortness of breath, pneumonia, diarrhea, renal failure, renal insufficiency, and septic shock.

6. A food composition for preventing or ameliorating Severe acute respiratory syndrome coronavirus 2 infectious diseases,

comprises the following steps:

a compound represented by the following chemical formula 1 or a bromatologically acceptable salt thereof; and

a compound represented by the following chemical formula 2 or a bromatologically acceptable salt thereof:

chemical formula 1:

chemical formula 2:

7. a composition for resisting severe acute respiratory syndrome coronavirus 2 is characterized in that,

comprises the following steps:

a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof; and

a compound represented by the following chemical formula 2:

chemical formula 1:

chemical formula 2:

8. a pharmaceutical composition for preventing or treating severe acute respiratory syndrome coronavirus 2 infectious diseases, characterized by,

comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof:

chemical formula 1:

9. the pharmaceutical composition for preventing or treating Severe acute respiratory syndrome coronavirus 2 infectious disease according to claim 8, wherein Severe acute respiratory syndrome coronavirus 2 infectious disease is a respiratory disease.

10. The pharmaceutical composition for preventing or treating Severe acute respiratory syndrome coronavirus 2 infectious disease according to claim 8, wherein Severe acute respiratory syndrome coronavirus 2 infectious disease shows symptoms after a latency period of 2 to 14 days after infection with said coronavirus.

11. The pharmaceutical composition for preventing or treating Severe acute respiratory syndrome coronavirus 2 infectious disease according to claim 8, wherein Severe acute respiratory syndrome coronavirus 2 infectious disease is a disease infected with Severe acute respiratory syndrome coronavirus 2, and shows one or more symptoms selected from the group consisting of hyperpyrexia, cough, shortness of breath, pneumonia, diarrhea, renal failure, renal insufficiency, and septic shock.

12. A composition for resisting severe acute respiratory syndrome coronavirus 2 is characterized in that,

comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof:

chemical formula 1:

13. a pharmaceutical composition for preventing or treating severe acute respiratory syndrome coronavirus 2 infectious diseases, characterized by,

comprising a compound represented by the following chemical formula 2:

chemical formula 2:

14. the pharmaceutical composition for preventing or treating Severe acute respiratory syndrome coronavirus 2 infectious disease according to claim 13, wherein Severe acute respiratory syndrome coronavirus 2 infectious disease is a respiratory disease.

15. The pharmaceutical composition for preventing or treating Severe acute respiratory syndrome coronavirus 2 infectious disease according to claim 13, wherein Severe acute respiratory syndrome coronavirus 2 infectious disease shows symptoms after a latency period of 2 to 14 days after infection with said coronavirus.

16. The pharmaceutical composition for preventing or treating Severe acute respiratory syndrome coronavirus 2 infectious disease according to claim 13, wherein Severe acute respiratory syndrome coronavirus 2 infectious disease is a disease infected with Severe acute respiratory syndrome coronavirus 2, and shows one or more symptoms selected from the group consisting of hyperpyrexia, cough, shortness of breath, pneumonia, diarrhea, renal failure, renal insufficiency, and septic shock.

17. A composition for resisting severe acute respiratory syndrome coronavirus 2 is characterized in that,

comprising a compound represented by the following chemical formula 2:

chemical formula 2:

18. a pharmaceutical composition comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof and a compound represented by the following chemical formula 2 or a pharmaceutically acceptable salt thereof for preventing or treating severe acute respiratory syndrome coronavirus 2 infectious diseases:

chemical formula 1:

chemical formula 2:

19. a method for treating severe acute respiratory syndrome coronavirus 2 infection, which is characterized in that,

comprising the step of administering a therapeutically effective amount of a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof and a compound represented by the following chemical formula 2 or a pharmaceutically acceptable salt thereof to a subject in need thereof:

chemical formula 1:

chemical formula 2:

20. use of a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof and a compound represented by the following chemical formula 2 or a pharmaceutically acceptable salt thereof for the preparation of a therapeutic agent for severe acute respiratory syndrome coronavirus 2 infection or a disease caused by severe acute respiratory syndrome coronavirus 2 infection:

chemical formula 1:

chemical formula 2:

Technical Field

The present invention relates to a composition for preventing or treating severe acute respiratory syndrome coronavirus 2 infection or a disease caused by severe acute respiratory syndrome coronavirus 2 infection.

Background

Severe acute respiratory syndrome coronavirus 2 is also known as COVID-19, SARS-CoV-2, 2019-nCoV virus. The World Health Organization (WHO) announced the international Public Health Emergency (PHEIC) for COVID-19 and announced COVID-19 as a global epidemic (pendemic) following hong kong flu (1968), influenza a H1N1 (2009) on day 11, 3/2020. COVID-19 is transmitted when droplets (saliva) from an infected person penetrate the mucous membranes of the respiratory tract or eyes, nose and mouth. In addition, when the infection is caused, respiratory symptoms such as fever, cough or dyspnea and main symptoms such as pneumonia occur after a latency period of 2 to 14 days (estimated), but asymptomatic infection cases also occur occasionally.

Coronaviruses are classified into α, β, γ and δ groups according to differences in RNA replication and transcriptase (RdRp) nucleic acid sequences. COVID-19virus is a virus that infects people in the beta group together with SARS-CoV and MERS-CoV virus (Subunit Vaccines Against infectious Pathogenic Human Coronavir. Wang N, Shang J, Jiang S, Du L. Frontnicrobiol.2020 Feb 28; 11: 298.), and the nucleic acid sequence of COVID-19 is 80% and 50% identical to SARS-CoV and MERS-CoV, respectively (Wu C. et al. analysis of thermal targets for SARS-CoV-2and discovery of genetic drugs by genetic computing methods.202Feb Acrmaceutica Sinica B).

Coronaviruses are positive single-stranded RNA viruses of 26-32kb in length, which, when introduced into a host through a host cell receptor, produce two large polymeric proteins: pp1a (486kDa), pp1ab (790kDa), thus using the protease of the virus itself: the 3CL protease (also known as 3CLpro or Mpro) produces 16 nonstructural proteins, including the polymerase (RdRp), Helicase (Helicase), ribonuclease (ribonuclease), and 3CLpro, etc. (Autoprocessing mechanism of viral transcription and replication) yield kinase 3C-like protease (SARS-CoV 3CLpro) fragments polyprotein. gamma. T, Kim YT, Nishii W, Terada T, Shirouzum, Yokoyama S.2013y; 280 (9): 2002-13. doi: 10.1111).

3CLpro is a protein (306 amino acids) with a molecular weight of 34kDa, and its initial abundance is reported to be automatically excised (automated processing) from pp1a, pp1ab (SARS-CoV 3CLpro) from peptides polypeptides. Muramatsu T, Kim YT, Nishii W, Terada T, Shirouzum, Yokoyama S.FEBS J.2013 may; 280 (9): 2002-13. doi: 10.1111). 3CLpro is a dimeric (dimer) structure consisting of domains I, II, III, with the active site (substrate binding site) located in the cleft (cleft) between domains I, II. The active site well preserves the amino acid sequence and three-dimensional structure between coronaviruses (96% of the structure is consistent with SARS-CoV). Recently, Jin Z.China, group reported the Structure of COVID-193 CLpro (Protein Data Bank ID 6LU7), wherein the sites showing structural differences among 12 coronavirus 3CLpro are domain III and surface loops (surface loops), and the active sites for preservation are high (Structure of from COVID-19 viruses and discovery of sites of interest in Jin Z, Du X, Xu Y, Deng Y, Lium, Zhao Y, Zhang B, Li X, Zhang L, Peng C, Duan Y, Yu J, Wang L, Yang K, Liu F, Jiang R, Yang X, You T, Liu X, Yax, Bai F, Liu H, Liu X, LW, Xu W, Xia G, Jian C, Yang Z, Yang H, J H, Yang H, J H, Yang K, Yang K H, Yang H, Na H3H, Na 3H, J, Na 3, K.

As described above, 3CLpro is one of important targets of therapeutic agents for coronavirus because the amino acid sequence and structure of 3CLpro are slightly changed among coronaviruses, and particularly, the function of 3CLpro is essential for virus propagation. Lopinavir (Lopinavir, phase 3: NCT04252274, NCT04251871, NCT04255017, ChiCTR2000029539), Ritonavir (Ritonavir, phase 3: NCT04251871, NCT04255017, NCT04261270), Darunavir (phase 3: NCT04252274), and the like, which are currently under development as COVID-19 therapeutics, are all targeted to 3CLpro, and these drugs were originally approved by the FDA as protease inhibitors of Human Immunodeficiency Virus (HIV: Human Immunodeficiency Virus) (Therapeutic for the 2019novel coronavir (2019-nCoV). Li, De Clercq E. Nat Rev Drug Discov.20280 (2020: 3) 149-150 i 10.1038).

As a technique related to a pharmaceutical composition for ameliorating or treating a coronavirus infection, korean laid-open patent No. 1913789 discloses a compound for treating a disease caused by a coronavirus infection, and U.S. published patent No. 2006-0257852 discloses a technique related to severe acute respiratory syndrome coronavirus, but does not disclose a composition for preventing or treating a coronavirus infection of the present invention, which comprises an active inhibitor of coronavirus 3CL protease as an effective ingredient.

Disclosure of Invention

An object of the present invention is to provide a pharmaceutical composition for preventing or treating severe acute respiratory syndrome coronavirus 2 infection disease, comprising a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

It is still another object of the present invention to provide a pharmaceutical composition for preventing or treating severe acute respiratory syndrome coronavirus 2 infection disease, comprising a compound represented by chemical formula 2 or a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating severe acute respiratory syndrome coronavirus 2 infection diseases, comprising a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof and a compound represented by chemical formula 2 or a pharmaceutically acceptable salt thereof as effective ingredients.

It is still another object of the present invention to provide a food composition for preventing or improving severe acute respiratory syndrome coronavirus 2 infection disease, comprising a compound represented by chemical formula 1 or a dietetically acceptable salt thereof.

It is still another object of the present invention to provide a food composition for preventing or improving severe acute respiratory syndrome coronavirus 2 infection disease, comprising a compound represented by chemical formula 2 or a dietetically acceptable salt thereof.

It is still another object of the present invention to provide a food composition for preventing or improving severe acute respiratory syndrome coronavirus 2 infection disease, comprising: a compound represented by chemical formula 1 or a bromatologically acceptable salt thereof; and a compound represented by chemical formula 2 or a dietetically acceptable salt thereof.

It is still another object of the present invention to provide a method for treating a severe acute respiratory syndrome coronavirus 2 infection disease, comprising the step of administering a therapeutically effective amount of a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof to a subject in need thereof.

It is still another object of the present invention to provide a method for treating a severe acute respiratory syndrome coronavirus 2 infection disease, comprising the step of administering a therapeutically effective amount of a compound represented by chemical formula 2 or a pharmaceutically acceptable salt thereof to a subject in need thereof.

It is still another object of the present invention to provide a method for treating a severe acute respiratory syndrome coronavirus 2 infection disease, which comprises the step of administering a therapeutically effective amount of a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof and a compound represented by chemical formula 2 or a pharmaceutically acceptable salt thereof to a subject in need thereof.

It is still another object of the present invention to provide an anti-severe acute respiratory syndrome coronavirus 2 composition comprising a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

It is still another object of the present invention to provide an anti-severe acute respiratory syndrome coronavirus 2 composition comprising a compound represented by chemical formula 2 or a pharmaceutically acceptable salt thereof as an active ingredient.

It is still another object of the present invention to provide an anti-severe acute respiratory syndrome coronavirus 2 composition comprising a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof and a compound represented by chemical formula 2 or a pharmaceutically acceptable salt thereof as effective ingredients.

It is still another object of the present invention to provide an inhibitor of the activity of 3CL protease, which comprises a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof.

It is still another object of the present invention to provide an inhibitor of the activity of 3CL protease, which comprises a compound represented by chemical formula 2 or a pharmaceutically acceptable salt thereof.

It is still another object of the present invention to provide an inhibitor of the activity of 3CL protease, comprising: a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof; and a compound represented by chemical formula 2 or a pharmaceutically acceptable salt thereof.

The present inventors have found a compound having a preventive or therapeutic effect on a severe acute respiratory syndrome coronavirus 2-infected disease and confirmed its inhibitory effect on severe acute respiratory syndrome coronavirus 2, thereby completing the present invention.

In one aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating severe acute respiratory syndrome coronavirus 2 infection diseases, comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

Chemical formula 1:

the compound represented by the above chemical formula 1 is Zafirlukast (Zafirlukast), which is known as cyclopentyl 3- {2-methoxy-4- [ (o-tolylsulfonyl) carbamoyl ] benzyl } -1-methyl-1H-indol-5-ylcarbamate (Cyclo pen 3- {2-methoxy-4- [ (o-tolylsulfonyl) carbamoyl ] benzyl } -1-methyl-1H-indol-5-ylcarbamate) by the International Union of Pure and Applied Chemistry (IUPAC).

In the present invention, the compound represented by the above chemical formula 1 can inhibit 3CL protease activity. Thus, the compound of the above chemical formula 1 can be applied to inhibit viral proliferation in host cells (individuals) that have been infected with severe acute respiratory syndrome coronavirus 2.

In still another aspect of the present invention, there is provided a pharmaceutical composition for preventing or treating severe acute respiratory syndrome coronavirus 2 infection diseases, comprising a compound represented by the following chemical formula 2 or a pharmaceutically acceptable salt thereof as an active ingredient.

Chemical formula 2:

the compound represented by the above chemical formula 2 is sulpirtone (sulfinyl), which is named 1, 2-diphenyl-4- [2- (phenylsulfinyl) ethyl ] pyrazolidine-3, 5-dione (1, 2-diphenyl-4- [2- (phenylsulfinyl) ethyl ] pyrazolidine-3, 5-dione) by the International Union of pure and applied chemistry. The above-mentioned sulpirenone is generally known as a platelet aggregation inhibitor, and its effect of inhibiting 3CL protease activity or anti-COVID-19 effect has not been disclosed yet.

In the present invention, the compound represented by the above chemical formula 2 can inhibit 3CL protease activity, and thus the compound of the above chemical formula 2 can be applied to inhibit viral proliferation in a host cell (individual) that has been infected with severe acute respiratory syndrome coronavirus 2.

In another aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating severe acute respiratory syndrome coronavirus 2 infection diseases, comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof and a compound represented by the following chemical formula 2 or a pharmaceutically acceptable salt thereof as effective ingredients.

Chemical formula 1:and

chemical formula 2:

in the present invention, the combination pharmaceutical composition comprising the compounds represented by the above chemical formulas 1 and 2 can inhibit 3CL protease activity. The compounds of the above chemical formulas 1 and 2 have a synergistic effect according to their combination, and can be applied to the inhibition of viral proliferation in host cells (individuals) infected with severe acute respiratory syndrome coronavirus 2.

In the present invention, the compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof and the compound represented by the above chemical formula 2 or a pharmaceutically acceptable salt thereof may be represented by 1: 99 to 99: 1, but is not limited thereto.

Also, in the present invention, the compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof and the compound represented by the above chemical formula 2 or a pharmaceutically acceptable salt thereof may be administered simultaneously, separately or sequentially in combination, and may be administered once or more times. In view of all of the above factors, it is important to administer the drug in an amount that can achieve the maximum effect at the minimum amount without side effects, which can be easily determined by one of ordinary skill in the art to which the present invention pertains.

Specifically, the composition comprising the compound of chemical formula 1, the composition comprising the compound of chemical formula 2, or the composition comprising the compounds of chemical formulae 1 and 2 according to the present invention can inhibit the protein segmentation step of virus-processing related proteins to proliferate in a subject after the subject is infected with severe acute respiratory syndrome coronavirus 2. This is different from inhibiting the activity of the first stage of viral infection, in particular from inhibiting the binding of viral host cell (individual) infection (viral entry) mechanisms.

Hereinafter, the present invention will be described in detail.

The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Unless a different meaning is explicitly stated in context, an expression in the singular includes an expression in the plural. In the present invention, the terms "comprises" or "comprising" are intended to specify the presence of stated features, steps, structures, or combinations thereof, but are not to be construed to preclude the presence or addition of one or more other features, steps, structures, or combinations thereof.

Unless defined otherwise, all terms including technical or scientific terms have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Terms commonly used such as terms defined in dictionaries should be interpreted as having a meaning that is consistent with a meaning that is possessed by context of the relevant art and should not be interpreted in an idealized or overly formal sense unless expressly so defined herein.

In the present invention, resistance to severe acute respiratory syndrome coronavirus 2 may represent preventive treatment or improvement of a severe acute respiratory syndrome coronavirus 2 infection disease, that is, severe acute respiratory syndrome coronavirus 2 infection or a disease caused thereby. Also, resistance to severe acute respiratory syndrome coronavirus 2 can be interpreted as all actions to reduce viral activity, including inhibition of proliferation, death, etc. of severe acute respiratory syndrome coronavirus 2 in a subject.

In the present specification, the above-mentioned "prevention" means that all behaviors of diseases caused by infection with severe acute respiratory syndrome coronavirus 2 or infection with severe acute respiratory syndrome coronavirus 2 are inhibited or delayed by administering the above-mentioned pharmaceutical composition. Also, the above "treatment" means to improve or beneficially modify all behaviors of disease symptoms caused by severe acute respiratory syndrome coronavirus 2 infection or by severe acute respiratory syndrome coronavirus 2 infection by administering the above pharmaceutical composition.

The disease caused by severe acute respiratory syndrome coronavirus 2 infection may be a respiratory disease. For example, the severe acute respiratory syndrome coronavirus 2-infected disease described above may develop symptoms after a latency period of 2 to 14 days after the viral infection. These symptoms include gastrointestinal symptoms such as hyperpyrexia, cough, shortness of breath, pneumonia, diarrhea, organ dysfunction (renal failure, renal dysfunction, etc.), septic shock, severe or even death, and include any symptoms caused by the viral infections described above.

The compound represented by the above chemical formula 1 or the compound represented by the above chemical formula 2 of the present invention includes not only pharmaceutically acceptable salts thereof but also isomers, hydrates and solvates which can be prepared by a conventional method.

In the present invention, pharmaceutically acceptable salts refer to salts commonly used in the pharmaceutical industry, including, for example: inorganic salts made from calcium, potassium, sodium, magnesium, etc.; inorganic acid salts made of hydrochloric acid, nitric acid, phosphoric acid, bromic acid, iodic acid, perchloric acid, sulfuric acid, and the like; organic acid salts prepared from acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid from acids, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid, and the like; sulfonates made from methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and naphthalenesulfonic acid; amino acid salts made of glycine, arginine, lysine, etc.; and amine salts made of trimethylamine, triethylamine, ammonia, pyridine, picoline, etc., but the kind of the salt represented in the present invention is not limited by the listed salts.

The above pharmaceutically acceptable salts may also be applied correspondingly to the bromatologically acceptable salts.

The pharmaceutical composition of the present invention may further comprise ingredients that do not increase the pharmaceutical effect but are generally used in pharmaceutical compositions to improve odor, taste, vision, etc. Also, the pharmaceutical composition of the present invention may further comprise pharmaceutically acceptable additives. Pharmaceutically acceptable additives include, for example, starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, lactose, mannitol, syrup, acacia, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, opal, sodium starch glycolate, carnauba lead, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, calcium stearate, white sugar, dextrose, sorbitol, talc, and the like, but are not limited thereto. In addition, the pharmaceutical composition may contain a substance having an activity against severe acute respiratory syndrome coronavirus 2 used alone or in the past.

The above-described pharmaceutical compositions of the present invention comprise a pharmaceutically acceptable carrier and may be formulated for oral or non-oral human or veterinary use. Diluents or excipients, such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants, may be used when formulating the pharmaceutical compositions of the present invention. Solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, and can be prepared by mixing at least one excipient such as starch, Calcium carbonate (Calcium carbonate), Sucrose (Sucrose) or Lactose (Lactose), gelatin and the like in a pharmaceutical composition containing the compound of the present invention. Besides simple excipients, lubricants such as magnesium, stearate, and talc may also be used. Liquid preparations for oral administration include suspensions, oral liquids, emulsions, syrups and the like, and may include various excipients such as moisturizers, sweeteners, aromatics, preservatives and the like, in addition to water and paraffin, which are generally used as simple diluents. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized formulations and suppositories. As the nonaqueous solvent and the suspending solvent, vegetable oils such as Propylene glycol (Propylene glycol), polyethylene glycol, and olive oil, and injectable esters such as ethyl oleate can be used. As the base of the suppository, there can be used vicisol (witepsol), polyethylene glycol, tween 61, cacao butter, laurate, glycerogelatin and the like.

The "hydrate" of the present invention means that in the compound represented by the above chemical formula 1 or the compound represented by the above chemical formula 2, water is non-covalently bonded by intermolecular force, and may include stoichiometric or non-stoichiometric water. Specifically, the above hydrate may include water in a ratio of about 0.25 mol to about 10 mol, more specifically, about 0.5 mol, about 1.5 mol, about 2 mol, about 2.5 mol, about 3 mol, etc., based on 1 mol of the active ingredient, but is not limited thereto.

The "solvate" of the present invention refers to a compound represented by the above chemical formula 1 or a compound represented by the above chemical formula 2 in which a solvent, not water, is bonded by intermolecular force, and may include stoichiometric or non-stoichiometric water. Specifically, the above hydrate may include water in a ratio of about 0.25 mol to about 10 mol, more specifically, about 0.5 mol, about 1.5 mol, about 2 mol, about 2.5 mol, about 3 mol, etc., based on 1 mol of the active ingredient, but is not limited thereto.

The pharmaceutical composition of the present invention may be administered orally or parenterally according to a desired method, and when the pharmaceutical composition is administered parenterally, it is preferable to select an injection form for external dermal or intraperitoneal injection, intrarectal injection, subcutaneous injection, intravenous injection, intramuscular injection, or intrapleural injection.

Also, the pharmaceutical compositions of the present invention may be administered by inhalation. It delivers the drug directly to the lungs, and is non-toxic and capable of exhibiting a longer duration of action in small doses. Administration by inhalation may be carried out using pharmaceutical preparations which can be inhaled via the airways, nasal cavity, etc., which may include inhalable particles or droplets containing the drug. Such inhalation administration is not limited thereto, and for example, one of a Dry Powder Inhaler (DPI) or a compressed metered dose inhaler (pMDI) may be used. The drug particles are lightly (lightly) compressed into a frangible (breakable) matrix contained, for example, inside a delivery device (dry powder inhaler). When operated, the delivery device abrades (abrads) some of the drug particles from the matrix and disperses them into the aspiration orifice which delivers the drug particles to the airway. Alternatively, the drug particles may be free flowing powder contained inside the reservoir of the delivery device (dry powder inhaler). The reservoir may be an integral chamber within the device, or a capsule, blister, or similar performing reservoir that is inserted into the device prior to manipulation. When operated, the device disperses a quantity of drug particles from the reservoir into the inhalation port which delivers the drug particles to the airway.

The above-mentioned pharmaceutical composition of the present invention can be administered to an individual for preventing or treating a disease caused by severe acute respiratory syndrome coronavirus 2 infection or severe acute respiratory syndrome coronavirus 2. The term "subject" or "subject" as used herein refers to mammals including horses, sheep, pigs, goats, dogs, etc., but preferably humans, suffering from a disease caused by severe acute respiratory syndrome coronavirus 2 infection or severe acute respiratory syndrome coronavirus 2, and suffering from a disease whose symptoms are ameliorated by administration of the above-described pharmaceutical composition of the present invention.

The term "administering" as used herein means introducing a pharmaceutical composition of the invention into an individual by any suitable method. The route of administration may be oral or non-oral by any of the usual routes to the target tissue. Also, the pharmaceutical compositions of the present invention can be administered by any device that can move them to the target cells.

The pharmaceutical compositions of the present invention are administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" as used in the present invention means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective amount level can be determined according to the body weight, sex, age, health condition, severity, pharmaceutical activity, sensitivity to drugs, administration time, administration route and excretion rate of a patient, treatment period, factors including drugs used simultaneously, and other factors well known in the medical field. The pharmaceutical composition of the present invention may be administered as a single therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with the conventional therapeutic agents, and may be administered once or more times. In view of all of the above factors, it is important to administer the drug in an amount that can achieve the maximum effect at the minimum amount without side effects, which can be easily determined by one of ordinary skill in the art to which the present invention pertains.

For example, in the pharmaceutical composition of the present invention, the compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof may be administered at a dose of 0.0001mg/kg to 500mg/kg, preferably at a dose of 0.001mg/kg to 100mg/kg, which can be administered once or in divided doses a day.

And, for example, in the pharmaceutical composition of the present invention, the compound represented by the above chemical formula 2 or a pharmaceutically acceptable salt thereof may be administered at a dose of 0.0001mg/kg to 500mg/kg, preferably at a dose of 0.001mg/kg to 100mg/kg, which can be administered once or in divided doses a day.

And, for example, in the pharmaceutical composition of the present invention, the compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof and the compound represented by the above chemical formula 2 or a pharmaceutically acceptable salt thereof may be administered in combination in the above-mentioned appropriate administration ranges.

The above-mentioned pharmaceutical composition of the present invention may further comprise one or more active ingredients exhibiting the same or similar drug effects, in addition to the compound represented by the above-mentioned chemical formula 1 and/or the compound represented by the above-mentioned chemical formula 2.

The present invention provides a method for preventing or treating severe acute respiratory syndrome coronavirus 2 infection or a disease caused by severe acute respiratory syndrome coronavirus 2 infection, comprising the step of administering a composition comprising a compound represented by the above chemical formula 1 in a therapeutically effective amount; a compound represented by the above chemical formula 2 or a combination of a compound represented by the above chemical formula 1 and a compound represented by the above chemical formula 2.

The term "therapeutically effective amount" used in the present invention refers to a compound represented by the above chemical formula 1, which is effective in preventing severe acute respiratory syndrome coronavirus 2 infection or a disease caused by severe acute respiratory syndrome coronavirus 2 infection; a combined amount of the compound represented by the above chemical formula 2 or a pharmaceutically acceptable salt thereof or the compound represented by the above chemical formula 1 and the compound represented by the above chemical formula 2.

The present invention provides a method for treating a severe acute respiratory syndrome coronavirus 2 infection disease, comprising the step of administering a therapeutically effective amount of a compound represented by the above chemical formula 2 or a combination of a compound represented by the above chemical formula 1 and a compound represented by the above chemical formula 2 to a subject in need thereof.

By administering a composition comprising the compound represented by the above chemical formula 1 or the compound represented by chemical formula 2, not only the disease itself is treated before the onset of symptoms, but also the symptoms thereof are suppressed or avoided. In the management of disease, the prophylactic or therapeutic dose of a particular active ingredient may vary depending on the nature (nature) and severity of the disease or condition and the route of administration of the active ingredient. The dosage and dosage frequency may vary according to the age, weight and response of the individual patient. One of ordinary skill in the art can readily select the appropriate dosage regimen taking these factors into account.

The subject may be a mammal including a human.

The above treatment method may further include contacting the compound represented by the above chemical formula 1; a compound represented by the above chemical formula 2; or a combination comprising a combination of the compound represented by the above chemical formula 1 and the compound represented by the above chemical formula 2, and an additional active agent which can exhibit a synergistic effect or an auxiliary effect together with the combination comprising the compound represented by the above chemical formula 1 or the compound represented by the above chemical formula 2, in a therapeutically effective amount, which is useful for treating diseases.

The present invention also provides a compound represented by the above chemical formula 1 for use in the preparation of a therapeutic agent for severe acute respiratory syndrome coronavirus 2 infection or a disease caused by severe acute respiratory syndrome coronavirus 2 infection; a compound represented by the above chemical formula 2; or a combination of the compound represented by the above chemical formula 1 and the compound represented by the above chemical formula 2. Acceptable adjuvants, diluents, carriers, etc. may be mixed with the compound represented by the above chemical formula 1 or 2 for preparing a medicament, and have a synergistic effect of active ingredients by making a complex formulation together with other active agents.

The present invention provides a compound represented by chemical formula 1 for treating severe acute respiratory syndrome coronavirus 2 infection disease; a compound represented by the above chemical formula 2; or a combination of the compound represented by the above chemical formula 1 and the compound represented by the above chemical formula 2.

The present invention provides a compound represented by the above chemical formula 1 for preventing or treating severe acute respiratory syndrome coronavirus 2 infection diseases; a compound represented by the above chemical formula 2; or a pharmaceutical composition comprising as active ingredients a compound represented by the above chemical formula 1 and a compound represented by the above chemical formula 2.

In still another aspect of the present invention, there is provided an anti-severe acute respiratory syndrome coronavirus 2 composition comprising a compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof as an effective ingredient.

In still another aspect of the present invention, there is provided an anti-severe acute respiratory syndrome coronavirus 2 composition comprising a compound represented by the above chemical formula 2 or a pharmaceutically acceptable salt thereof as an active ingredient.

In still another aspect of the present invention, there is provided an anti-severe acute respiratory syndrome coronavirus 2 composition comprising a compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof and a compound represented by the above chemical formula 2 or a pharmaceutically acceptable salt thereof as effective ingredients.

In still another aspect of the present invention, there is provided an inhibitor of the activity of 3CL protease, comprising a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof.

In still another aspect of the present invention, there is provided an inhibitor of the activity of 3CL protease, comprising a compound represented by chemical formula 2 or a pharmaceutically acceptable salt thereof.

In still another aspect of the present invention, there is provided an inhibitor of the activity of 3CL protease, which comprises a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof and a compound represented by chemical formula 2 or a pharmaceutically acceptable salt thereof.

Since the amino acid sequence and structure of 3CLpro have little variation among coronaviruses, particularly the function of 3CLpro is essential for virus propagation, 3CLpro can be used as one of important targets of a coronavirus therapeutic agent.

In the present invention, a compound represented by the above chemical formula 1; a compound represented by the above chemical formula 2; or a combination of the compound represented by the above chemical formula 1 and the compound represented by the above chemical formula 2 has an excellent inhibitory effect on the activity of 3CL protease.

As can be confirmed from the above, zafirlukast, sulpirenone, and zafirlukast and sulpirenone according to the present invention can exhibit a very excellent ameliorating effect on the lung disease rate, and thus can exhibit a rapid treatment and ameliorating effect on infection.

Unless contradicted by each other, the same applies to the items mentioned in the use, composition, treatment method of the present invention.

In still another aspect of the present invention, the present invention provides a food composition for preventing or improving severe acute respiratory syndrome coronavirus 2 infection disease, comprising a compound represented by the following chemical formula 1 or a dietetically acceptable salt thereof.

Chemical formula 1:

in still another aspect of the present invention, the present invention provides a food composition for preventing or improving severe acute respiratory syndrome coronavirus 2 infection disease, comprising a compound represented by the following chemical formula 2 or a dietetically acceptable salt thereof.

Chemical formula 2:

in still another aspect of the present invention, there is provided a food composition for preventing or improving severe acute respiratory syndrome coronavirus 2 infection disease, comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof, and a compound represented by the following chemical formula 2 or a pharmaceutically acceptable salt thereof.

Chemical formula 1:and

chemical formula 2:

the description in the above pharmaceutical composition can be applied to the compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof and the compound represented by the above chemical formula 2 or a pharmaceutically acceptable salt thereof.

In the present invention, the above-mentioned food composition may be used together with other foods or food ingredients as it is, and may be suitably used according to a conventional method.

In the present specification, the term "food composition" refers to a food prepared and processed using a raw material or an ingredient having a functional property useful for a human body.

The kind of the above-mentioned food is not particularly limited. Examples of foods that can be added to the composition comprising the compound represented by the above chemical formula 1 or a dietetically acceptable salt thereof include various soups, drinks, teas, oral liquids, alcoholic beverages, vitamin complexes, etc., and may include all health foods in the conventional sense. In addition to the above, the composition of the present invention, which includes the compound represented by the above chemical formula 1 or a dietetically acceptable salt thereof, may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectin and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, ethanol, carbonating agents used in carbonated beverages, and the like. In addition, the food composition of the present invention may contain pulp for preparing natural fruit juice, fruit juice beverages and vegetable beverages. These components may be used alone or in admixture thereof.

The pharmaceutical composition of the present invention has the effect of preventing or treating a disease caused by severe acute respiratory syndrome coronavirus 2 infection or severe acute respiratory syndrome coronavirus 2.

The food composition of the present invention has the effect of preventing or improving a disease caused by severe acute respiratory syndrome coronavirus 2 infection or severe acute respiratory syndrome coronavirus 2.

Drawings

FIG. 1 shows AI-based New drug platform (DeepMatcher)^TM) The AI deep learning algorithm of (1).

Fig. 2 is a graph showing the results of cytotoxicity evaluation and cytopathic effect evaluation of the compounds of chemical formula 1 and chemical formula 2 according to the present invention.

Fig. 3 shows the observation results of cell morphology before and after treatment with the compounds of chemical formula 1 and chemical formula 2 of the present invention.

Fig. 4 shows the results of 3CL protease activity of the compounds of chemical formula 1 and chemical formula 2 according to the present invention.

FIG. 5 shows the results of evaluating the drug efficacy of ACE 2and SARS-CoV-2Spike S1 protein binding activities of the compounds of chemical formulas 1 and 2 of the present invention.

Fig. 6 shows a baseline Lung map for scoring (r) to (v) according to Lung lobes (Lung lobes) and assessing Lung morbidity by weight.

Fig. 7 is a graph showing the results of confirmation of the macroscopic treatment rate, in which NC (Negative control), VC (Virus control), PC (Positive control), rdsievir (Remdesivir), Za (Zafirlukast), Za + Sulf (Zafirlukast and sulpirenone).

Figure 8 shows the macroscopic lung disease rates confirmed by performing the experiments.

Detailed Description

The embodiments are presented to facilitate an understanding of the invention. The following examples are provided only to facilitate understanding of the present invention, and the present disclosure is not limited to the examples.

Preparation example 1 Zafirlukast (Zafirlukast)

Commercially available zafirlukast (CAS Number: 107753-78-6) was used.

Preparation example 2 Sulfopyrone (Sulfinpyrazone)

Commercially available sulpirtone (CAS Number: 57-96-5) was used.

Preparation example 3 Combined use of zafirlukast and Sulfpirone

The zafirlukast and the sulpirtone are used in combination.

EXAMPLE 1 screening of candidate Compounds that inhibit COVID-193 CL protease

In The present invention, The Protein-ligand complex structure information based on The COVID-193 CL protease (Protein Data Bank ID 6LU 7: The crystal structure of COVID-19main protease in complex with an inhibitor N3, DOI: 10.2210/pdb6LU7/pdb) was passed through an AI-based New drug platform (DeepMatcher)^TM) The AI deep learning algorithm of (a) from 2700 multiple FDA approved drugs (source: https: // www.drugbank.ca /) were searched for 3CL protease inhibiting compounds and lead out drug candidates. AI-based new drug platform (DeepMatcher) for use in the invention^TM) The 3D-virtual graph simulation method (2) is a technique that can simulate the interaction of a protein-ligand complex according to changes in the actual physical environment (changes in the structure, state, and physical properties of a molecule), and can analyze and measure the number of moving snapshots of a drug molecule bound to the binding site of a protein to thereby simulate the binding frequency (high binding quality) of the proteinThe index (HBQI: high binding quality index)) and the binding structure similarity (RMSD: root mean Square Deviation) and the like (FIG. 1).

EXAMPLE 2 evaluation of SARS CoV-2 antiviral (cytopathic inhibitory Effect) drugs

Material (drug): zafirlukast and sulpirtone

Test viruses and cell lines: SARS CoV-2(NCCP-43326), VERO cell (cell)

Assay concentration and number of replicates: 2 doses (Dose) (10-20. mu.M)/repeat 2 times

Example 2-1: cytotoxicity assay (evaluation of cytotoxicity)

DMEM (5% FBS, 1% antimicrobial-antimicrobial) was used as cell culture medium and the test samples (drugs) were evaluated for cytotoxicity by MTT assay (MTT assay). The final concentration of the solvent (DMSO) used when preparing samples of different test concentrations was 0.5% or less.

Cytotoxicity evaluation was performed in the following order.

1) At a rate of 5X 10 per hole⁴Cells/well (cells/well) were aliquoted into 96-well plates (well plates) at 37 ℃ with 5% CO²After 48 hours of incubation in the incubator, cell monolayers were obtained and then washed 2 times with PBS.

2) After 100. mu.L/well of the samples (drugs) prepared at different test concentrations were aliquoted (including the culture broth), 5% CO was added at 37 ℃²The culture was carried out in an incubator for 72 hours.

3) After confirming the cell status, MTT solution was added at 10. mu.L/well, followed by 5% CO at 37 ℃²The incubator was allowed to stand for 4 hours.

4) After 4 hours of reaction, MTT solution (solution) was added at 100. mu.L/well, then formazan (formazan) crystals were sufficiently dissolved using the precipitate, and absorbance was measured at 570nm with a microplate reader.

5) The cytotoxicity ratio was calculated according to the following formula 1 based on the ratio of normal cells and the group treated with the test sample (drug).

Mathematical formula 1

Cell Viability (Cell Viability) (%) Test OD/Control OD × 100%

Example 2-2: CPE Reduction Test (cytopathic inhibition rate) (evaluation of cytopathic inhibition efficacy)

Evaluation of the cytopathic inhibitory effect was performed in the following manner.

1) At a rate of 5X 10 per hole⁴After cell/well plating to 96-well plates, 5% CO was added at 37 deg.C²After 48 hours of culture in the incubator, a cell monolayer was obtained, followed by washing 2 times with PBS and cell counting.

2) After 100. mu.L/well of DMEM (FBS-free, 1% antibiotic-antifungal) medium was used for strain division, 5% CO at 37 ℃²The incubator was allowed to stand for 1 hour to allow infection of the virus at 0.001 MOI.

3) 1 hour after infection, the virus was removed, and then samples (drugs) prepared at different test concentrations were aliquoted (including culture broth) at 100. mu.L/well and then subjected to 5% CO at 37 ℃ C²The culture was carried out in an incubator for 72 hours.

4) After confirming the cell status, MTT solution was added at 10. mu.L/well, followed by 5% CO at 37 ℃²The incubator was allowed to stand for 4 hours.

5) After 4 hours of reaction, MTT solution (solution) was added at 100. mu.L/well, then formazan (formazan) crystals were sufficiently dissolved using the precipitate, and absorbance was measured at 570nm with a microplate reader.

6) The cytotoxicity ratio was calculated according to the following formula 2 based on the ratio of normal cells and the group treated with the test sample (drug).

Mathematical formula 2

Virus inhibition rate(％)＝(Test OD-Virus OD)/(Control OD-Virus OD)×100％

The evaluation results of the cytotoxicity-based anti-COVID-19 efficacy confirmed according to the above examples 2-1 and 2-2 are shown in the following Table 1 and FIG. 2.

TABLE 1

As a result of evaluating the anti-COVID-19 efficacy, it was confirmed that both zafirlukast and/or sulpirenone drugs had excellent cell survival rates, and that zafirlukast had a cytopathic effect of 75.6 ± 3.9% and sulpirenone had a cytopathic effect of 40.7 ± 8.0% based on 20 μ M, as shown in table 1 and fig. 2.

Example 3 Observation of cell morphology based on drug treatment

Material (drug): zafirlukast and sulpirtone

Test viruses and cell lines: SARS CoV-2(NCCP-43326), VERO cell (cell)

Assay concentration and number of replicates: 2 doses (10-20. mu.M)/repeat 2 times

The morphological changes of the cells occurring when treated with the drug were observed for each of the cells before and after the treatment with the virus, and the results thereof are shown in fig. 3.

As confirmed in fig. 3, in the control group, it was confirmed that the morphological change of the Virus-infected cells (Virus) was larger than that of the Virus-uninfected cells (Nomal).

In contrast, it was confirmed that the morphological change of each experimental group treated with zafirlukast and sulpirenone on virus-infected cells was small.

Example 4.3CL protease (3-chorymopepsin-like protease) inhibition efficacy assay (assay)

Material (drug): zafirlukast and sulpirtone

Assay 3CL Protease:

GC376 is purchased from Aobious(catalog number AOB36447)

3CL Protease assay (3CL Protease assay) Buffer (BPS catalog number 79956)

3CL Protease Substrate(BPS catalog number 79952)

Assay concentration and number of replicates: 10 doses (0.001-30 μ M)/repeat 2 times

The analysis of the efficacy of 3CL Protease inhibition using the time-lapse fluorescence resonance energy transfer assay was performed as follows.

1) Each drug was used dissolved in 100% DMSO at a concentration of 10mM, and an intermediate dilution process was performed while maintaining the 100% DMSO concentration.

2) After the compound was diluted 67.5 times with 3CL Protease assay (3CL Protease assay) buffer, 50ul of a reaction solution having a DMSO concentration of 0.3% was prepared using 10ul of the diluted sample and 40ul of the buffer.

3) After allowing each drug to stand at room temperature for 30 minutes, the drugs were mixed so that the final concentration of the substrate for the 3CL protease reached 50 uM.

4) The reaction mixture was allowed to react at room temperature for 12 hours or more.

5) The TR-FRET signal is then read from an Infinitem1000 plate reader (Tecan).

The results of evaluating the efficacy of the SARS-CoV-23 CL protease protein activity confirmed by the above are shown in Table 2, Table 3 and FIG. 4 below.

TABLE 2

TABLE 3

Referring to tables 2and 3 and fig. 4, it was confirmed that zafirlukast at a concentration of 30 μ M inhibited the 3CL protease by about 45% or more, and sulpirenone at a concentration of 30 μ M inhibited the 3CL protease by about 20%. Therefore, it was confirmed that zafirlukast and/or sulpirenone had an inhibitory effect on the activity of 3CL protease (3-chymotryptsin-like protease).

Example 5. preparation of the compound by CE 2: spike S1 time-lapse fluorescence resonance energy transfer analysis for evaluating the efficacy of drugs on the binding activity of ACE 2and SARS-CoV-2Spike S1 proteins

Material (drug): zafirlukast and sulpirtone

Test ACE 2and Spike S1 proteins:

ACE2，His-Tag，Eu-labeled(BPS Bioscience，100705)

Spike S1，Fc fusion，Avi-tag，Biotin Labeled(SARS-CoV-2)(BPS Bioscience，100679)

Anti-Spike，Neutralizing Antibody(Activemotif，91361)

assay concentration and number of replicates: ACE2 was performed at 10 doses (0.002-30 μ M) in 2 replicates as follows: spike S1 time-lapse fluorescence resonance energy transfer analysis.

1) After diluting ACE2-Eu to 1ng/uL (12nm) using 1 XACE 2-Spike TR-FRET buffer, it was aliquoted into each well in an amount of 5 uL.

2) After diluting Dye-labeled (Dye-labeled) acceptors 100-fold using 1 × ACE2-Spike TR-FRET buffer, the diluted Dye-labeled acceptors were added to each well in an amount of 5 uL.

3) Samples (drugs) prepared at different concentrations were added to the wells in an amount of 5 uL.

4) After diluting Spike S1-Biotin to 20ng/uL (200nm) using 1 XACE 2-Spike TR-FRET buffer, the diluted Spike S1-Biotin was added in an amount of 5uL to wells designated as test and control groups.

5) 1x ACE2-Spike TR-FRET buffer was added to the remaining wells without addition in an amount of 5uL and the plates were incubated for 1 hour at room temperature.

6) The TR-FRET signal was then read from an Infinite M1000 microplate reader (Tecan).

The results of evaluating the pharmaceutical efficacy of the binding activity of ACE 2and SARS-CoV-2Spike S1 protein thus confirmed are shown in Table 4, Table 5 and FIG. 5 below.

TABLE 4

TABLE 5

Referring to table 4, table 5 and fig. 5 above, it was confirmed that zafirlukast and sulpirenone had no inhibitory effect on the binding of ACE 2and Spike S1 proteins. That is, it was confirmed that zafirlukast and sulpirenone are not drugs which exert their effects by binding to ACE 2and Spike S1, but drugs which exhibit their effects of treating diseases by inhibiting 3CL protease.

Example 6 evaluation of drug efficacy in animal models infected with Severe acute respiratory syndrome coronavirus 2 Using Syrian Hamster (Syrian Hamster)

The therapeutic effect on severe acute respiratory syndrome coronavirus 2 infection was confirmed using a syrian hamster animal model.

The experiment was carried out according to the conditions of table 6 below.

TABLE 6

Hamster (Hamsters)	Female (Female), 5n/groups
		Age (Age)	About 1 month old
Virus Titer (Virus Titer)	2.0×105PFU/mL
		Route of inoculation (Inoculate Route)	Intranasal (Intranasal) (100. mu.L)
Administration (Drug Administration)	Oral (per oral) (PO, 500 μ L)

Experiments were performed by administering severe acute respiratory syndrome coronavirus 2 according to the above conditions. The conditions for the test groups are given in table 7 below.

TABLE 7

The experimental apparatus was configured according to the above conditions.

After a 10-minute respiratory anesthesia using Isoflurane (Isoflurane) and 100. mu.L of prepared SARS-Cov-2 virus was instilled into the nasal cavity of hamster to infect the hamster, 500. mu.L of the prepared drug was orally administered to the hamster using a probe. On day 4 post-infection (PID4), lung tissue collected by autopsy was evaluated for gross lung disease rates. The macroscopic lung lesion rate was calculated from the ratio of the virus group to the test group by the following formula.

Specifically, as shown in fig. 6, the lungs were classified into (r) to (c) according to the lobes of the lungs (Lung), and the Lung morbidity rate was evaluated according to the weight of each fraction. The criteria are shown in table 8 below.

TABLE 8

Lung lobe (Lung lobe)	Percentage of lung weight (% of Lung weight) (%)
		①	40
②	10
		③	5
④	30
		⑤	15
Total area (Total area)%	100

The visual treatment rate was evaluated according to the criteria of table 8, and the result value was confirmed by the following formula, and the result is shown in fig. 7.

Mathematical formula 3

Piperaceae 100 for treating pneumonia by naked eye

Figure 7 shows the macroscopic treatment rate. As can be confirmed from fig. 7, the treatment efficacy for about 5 days after continuous administration as compared with the severe acute respiratory syndrome coronavirus 2 alone infection group (severe infection model, lung disease variability 62%) was as follows: it was confirmed that the therapeutic rate of zafirlukast (60mg/kg/day) + sulpirenone (120mg/kg/day) was 94.3%, and the therapeutic rate of zafirlukast (60mg/kg/day) was 58.7%. On the other hand, Reidesvir (Remdesivir) (5mg/kg/day) as a control drug showed a treatment rate of 44.3%. From the above, it was confirmed that zafirlukast, and sulpirenone according to the present invention exhibited significantly superior therapeutic effects as compared to ridciclovir, which is a control drug. And zafirlukast, and sulpirtone all show excellent improvement effects in bilateral pneumonia, pulmonary edema, and pulmonary hemorrhage, and particularly zafirlukast and sulpirtone show the most excellent treatment efficacy clinically, and no abnormal findings or abnormal clinical symptoms are observed.

Also, the macroscopic lung disease rate confirmed by performing the above experiment is specifically shown in fig. 8. Abbreviations in fig. 8 are as follows: NC (Negative control), VC (Virus control), PC (Positive control), Za (zafirlukast), Z + S (zafirlukast and sulpirenone).

From the above, it was confirmed that Za (zafirlukast), Z + S (zafirlukast and sulpirenone) of the present invention exhibited very excellent improving effects in terms of macroscopic lung disease rate. In particular, it was confirmed that zafirlukast and sulpirenone exhibited very excellent improving effects up to the level close to the normal control group.

Example 7 evaluation of drug efficacy of Sulfpyrone in animal models infected with Severe acute respiratory syndrome coronavirus 2 Using Syrian hamster

The therapeutic effect of sulpirenone on severe acute respiratory syndrome coronavirus 2 infection was confirmed using a syrian hamster animal model.

The experiment was carried out according to the conditions of table 9 below.

TABLE 9

Hamster (Hamsters)	Female (Female), 5n/groups
		Age (Age)	About 1 month old
Virus Titer (Virus Titer)	2.0×105PFU/mL
		Route of inoculation (Inoculate Route)	Intranasal (Intranasal) (100. mu.L)
Administration (Drug Administration)	Oral (per oral) (PO, 500 μ L)

Experiments were performed by administering severe acute respiratory syndrome coronavirus 2 according to the above conditions. The conditions for the test groups are given in table 10 below.

Watch 10

The experimental set-up was configured according to the above conditions, and the lung disease rate was evaluated, and the macroscopic treatment rate evaluation method was performed in the same manner as that performed in the examples, and the results thereof are shown in the following Table 11.

TABLE 11

From the results of the macroscopic therapeutic rate analysis in FIG. 11, it was confirmed that the therapeutic rate of sulpirtone (50mg/kg/day) was 15.6% as a result of the therapeutic efficacy of continuous administration for about 4 days, as compared with the severe acute respiratory syndrome coronavirus 2 alone-infected group.

From the above, it was confirmed that the sulpirtone of the present invention showed an improved effect in terms of macroscopic treatment rate.

A detailed description of what can be fully recognized and analogized by those skilled in the art to which the present invention pertains is omitted in the present specification, and various modifications may be made within a scope not changing the technical idea or essential structure of the present invention, in addition to the specific examples described in the present specification. Accordingly, the present invention may be embodied in other ways than those specifically described and illustrated herein, as would be understood by one of ordinary skill in the art to which the present invention pertains.