WO2021250606A1

WO2021250606A1 - Composition for treating coronavirus disease-19 (covid-19)

Info

Publication number: WO2021250606A1
Application number: PCT/IB2021/055105
Authority: WO
Inventors: 강진석; 윤치호; 유안나; 김준영
Original assignee: 제이더블유중외제약 주식회사
Priority date: 2020-06-11
Filing date: 2021-06-10
Publication date: 2021-12-16
Also published as: KR20210153908A; KR20230026396A

Abstract

The present invention relates to a treatment composition comprising a compound capable of very specifically and highly efficiently inhibiting severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2).

Description

Coronavirus infection ^^( )^!)-:! ⁹ ) Field of invention of therapeutic composition The present invention relates to a composition for treating coronavirus infection-19 comprising a compound capable of inhibiting SARS-CoV-2 (Severe acute respiratory syndrome coronavirus 2) with very specific and high efficiency. . Background Art Coronavirus (coronavirus) is a kind of RNA virus, genetic information is a virus consisting of ribonucleic acid (RNA), it causes respiratory and digestive system infections in humans and animals. Mainly, it is easily infected by mucosal infection or droplet transmission, and healthy people usually cause mild respiratory infections, but rarely cause fatal infections. Among coronavirus infections, the severe acute respiratory syndrome-coronavirus (SARS-CoV), which broke out from China in April 2003, recorded a mortality rate of 9.6% and killed many people, and in 2015 the Middle East respiratory syndrome-coronavirus (MERS-) CoV) originated from the Middle East and spread worldwide, showing a high mortality rate of about 36%, and the severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) epidemic in December 2019 from China is currently ongoing. is in progress Severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) first appeared in Wuhan, China and has spread rapidly worldwide, and the WHO named the disease infected with the virus COVID-19. According to a recent report, the general symptoms of fever (Lam ^0.6%), fatigue ⁽⁶⁹ 0.6%), dry cough ⁽⁵ ^9.4%), lymphopenia ⁽⁷ ^0.3%), extension of the prothrombin time (58 %), and enhancement of lactate dehydrogenase (39.9%) (Wang, D. ei., JAMA, 2020). It has been reported that the SARS-CoV-2 is transmitted mainly through human-to-human contact through respiratory droplets produced by coughing and sneezing or through surfaces of objects contaminated by people who cough or sneeze (CDC, How COVID-19 Spreads. Coronavirus Disease 2019 (COVID-19), 2020), asymptomatic infection is reported as possible

(Yu, P. et al., J Infect Dis, 2020; Hoehl, S. et al., N Engl J Med, 2020; Bendix, A., Science Alert, 2020).

As cases of SARS-CoV-2 surge worldwide, WHO declared a 'Public Health Emergency of International Concern' (PHEIC) on January 30, 2020. On the 11th, after Hong Kong Flu (1968) and H1N1 Flu (2009), Corona 19 was declared a pandemic (global pandemic) for the third time in history. As of June 2021, approximately 170 million cases have occurred worldwide, approximately 3,547,000 people have died, and tens of thousands of confirmed cases are added every day, and the number of infected patients is still on the rise. . In Korea, about 140,000 patients have been confirmed,

1,963 deaths have been reported (COVID-19 Dashboard by the Center for Systems Science and Engineering (CSSE) at Johns Hopkins University (JHU), Korea Centers for Disease Control and Prevention website) X Despite this global pandemic and severity, currently other viruses ( Influenza, Ebola virus, etc. (National Library of Medicine. Clinical Features of Suspected and Confirmed Patients of 2019 Novel Coronavirus Infection) Since comprehensive treatment as described above is only a potential treatment method using agents that have not proven specific for coronavirus, the patient’s The clinical effect is very different for each patient depending on the characteristics, immunity and condition, and the therapeutic effect is small, especially for patients with underlying diseases or the elderly with weak immunity, including children and the elderly, COVID-19 There is a need to develop a treatment that has been clinically proven to have a specific therapeutic effect.Currently, research on a treatment for COVID-19 based on an antiviral that has been approved as a treatment for other viral infections is in progress, specifically indinavir and saquina. Antiretroviral human immunodeficiency I protease (HIV-1 protease) inhibitors such as saquinavir, lopinavir/ritonavir, RNA polymerase inhibitors such as remdesivir ( polymerase inhibitor), investigation against Lhasa fever and Ebola strep fever The new drug triazavirin, the antiviral protein erdnterferon beta, the previously identified monoclonal antibody (Monoclonal) antibody), etc., are under research or clinical trials as candidates for treatment for COVID-19, but currently approved treatments for COVID-19 are Remdesivir (Gilliad Sciences) and Sotrobi, which have been approved by the US FDA. There is Mab (GSK), and currently approved treatments in Korea are Gilead Sciences' remdesivir and Celltrion's lekkonase (Paules, Catharine I., Marston, Hilary D., Fauci, Anthony S. (2020.01) .23).,

Coronavirus Infections - More Than Just the Common Cold; "Ural scientists have proposed testing the drug against coronavirus. Ural Federal University, 23/1/2020", etc.). Even now, a number of pharmaceutical companies around the world are making efforts to develop vaccines, but vaccines or therapeutic agents that show satisfactory therapeutic or preventive effects have not been reported yet, and there is still an urgent need for therapeutic agents with excellent therapeutic and preventive effects. to be. Under this background, the present inventors confirmed that a compound having an existing anticancer effect can inhibit SARS-Cov-2 with very specific and high efficiency, and thus the compounds can be used for the treatment of coronavirus infection-19 and completed the present invention.

【Prior art literature】

【Patent Literature】

(Patent Document 1) Republic of Korea Patent No. 1,692, 921 [Non-Patent Document]

(Non-Patent Document 1) 'Health and Welfare Issue & Focus' No. 373 issued by the Korea Institute for Health and Social Affairs (2020. 4.)

(Non-Patent Document 2) Seongsil Moon, BRIC View 2020-TX2 (2020) Summary of the Invention An object of the present invention is to provide a pharmaceutical composition for the treatment of coronavirus infection-19 or a pharmaceutical formulation comprising the same. Another object of the present invention is to provide a pharmaceutical composition for the treatment of coronavirus infection-19 or a pharmaceutical formulation comprising the same for treatment of coronavirus infection-19. 2021/250606 - €1/162021/055105 To provide a method of treating coronavirus infection-19, comprising administering to a subject in need. In order to achieve the above object, the present invention provides a pharmaceutical composition for treating coronavirus infection-19 comprising a compound having the structure of Formula I, a prodrug thereof, a salt thereof, or an isomer thereof.

In Formula I, ¾ it is (: 1, and the alkynyl group of the alkenyl group, or 02不₆ of不₆ alkyl group, 02不₆ of, ¾ an aryl group, a substituted aryl group, or a - (: (= 0) ¾ and ¾ is

Alkyl group, an alkynyl group of 02不₆ alkenyl group, or 02不_6. In the compound of Formula I included in the pharmaceutical composition for the treatment of coronavirus infection-19 according to the present invention, ¾ is an alkyl group of _1不6 _{or an alkenyl group of 02不6} , and ¾ is -(:(=0) ) ¾, ¾ is (: _{preferably an alkyl group of 1不6} , ¾ is methyl), and ¾ is more preferably -(: (=0) (: ¾), but is not limited thereto. In the compound of formula (I), ¾ is methyl (11), and ¾ is -(: (=0) (: ¾), has the structure of formula (II).

[Formula II]

In addition, the prodrug of the compound of formula (I) according to the present invention has the structure of formula (III). 2021/250606 1^(:1^2021/055105

[Formula III]

In Formula III, it is ¾ (in: 1不₆ alkyl group, 02不₆ alkenyl group, or ₆ of the 02不

An alkyl group, an alkenyl group of _02不6 , or an alkynyl group of _02不6 ^{, ¾ is -1 >} 0 ₃ ¾ -When 0 ₃ - ' -

1 ^> 0 ₃ ²明 2 ⁺ , -1 ^> 0 ₃ ² ¾ ₂ ⁺ ，- 1 ^> 0 ₃ ² -1^句²⁺ , -1 ^> 0 ₃ ^{2 2+} , Structural Formula 1, Structural Formula 2, or Structural Formula 3 to be.

In particular, in the formula (III) according to the invention compound, Ra is a C1-C alkyl group or alkenyl group of C2-C ₆ of _6, R _b is -C (= 0) Re, Re is a C _l -C ₆ Alkyl group, Rp is preferably -PO ₃ H2, -HP0 ₃ ^' Na ⁺ or -P0 ₃ ² -Na2, Ra is methyl (methyl), ¾ is -

C(=0)CH ₃ , Rp _{is more preferably -PO 3} H2, -HP0 ₃ ^' Na ⁺ or -P0 ₃ ² _Na2, but is not limited thereto. Most preferably, the prodrug of the compound of formula (I) according to the present invention, formula (III), may have the structures of formula (IV) to formula (VI), but is not limited thereto. The compounds having the structures of Formulas I to VI according to the present invention may be prepared by the method described in Korean Patent No. 1,692, 921, but is not limited thereto. 2021/250606 ?€1/162021/055105

The present invention also relates to pharmaceutically acceptable salts of compounds of formulas (I) to (VI). Non-limiting examples of pharmaceutically acceptable salts of compounds of Formulas I-VI include hydrochloric acid, !)-toluenesulfonic acid, fumaric acid, citric acid, succinic acid, salicylic acid, oxalic acid, hydrobromic acid, phosphoric acid, methanesulfonic acid, tartaric acid, horse salts of lactate, di-toluoyl tartaric acid, ortinic acid, edicylic acid, hemi-edicylic acid and mandelic acid may be exemplified, but not limited thereto. One or more pharmaceutically acceptable carriers may be included in the therapeutic composition comprising the compounds of formulas (I) to (VI) according to the present invention. The therapeutic composition according to the present invention may be administered orally or parenterally. For parenteral administration, formulations suitable for intranasal, intranasal, buccal, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, enteral, topical, sublingual or rectal dosage forms may be prepared and administered by the above route, but is not limited thereto. Accordingly, the present invention provides a pharmaceutical formulation for oral or parenteral administration comprising the therapeutic composition according to the present invention. Preferably, the therapeutic composition according to the present invention or a pharmaceutical formulation comprising the same may be administered orally, or may be administered intranasally or intranasally, particularly in the form of a spray or aerosol when administered intranasally or intranasally. It is more preferably formulated and administered in a formulation for administration or inhalation administration, but is not limited thereto. The pharmaceutical formulation for intranasal or intranasal administration may be prepared according to techniques well known in the art, and benzyl alcohol or other suitable preservatives, absorption enhancers to enhance bioavailability, fluorocarbons and/or others It can be prepared as a solution in saline using solubilizing or dispersing agents known in the art. In another form, the therapeutic composition according to the present invention may be formulated in a sterile injectable formulation as a sterile injectable aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents (eg, Tween 80) and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension (eg, a solution in 1,3-butanediol) in a non-toxic parenterally acceptable diluent or solvent. Non-limiting examples of pharmaceutically acceptable vehicles and solvents include mannitol, water, Ringel's solution, and isotonic sodium chloride solution. In addition, sterile, non-volatile oils are conventionally employed as the solvent or suspending medium. For this purpose, any non-volatile, less irritating oil may be used, including synthetic mono or diglycerides. Fatty acids such as oleic acid and its glyceride derivatives are useful in injectable formulations as are pharmaceutically acceptable natural oils (eg olive oil or castor oil), especially their polyoxyethylated ones. In addition, the therapeutic composition according to the present invention may include any orally acceptable composition including, but not limited to, capsules, pellets, tablets, aqueous suspensions and solutions. 2021/250606 1^(:1^2021/055105 It may be formulated in the form of a pharmaceutical dosage form and administered orally. When formulated for oral administration, one pharmaceutically acceptable one in the therapeutic composition according to the present invention It may be formulated to include the above excipients, and such excipients include fillers (diluents), disintegrants, binders, lubricants (lubricants), preservatives, antioxidants, buffers, chelating agents, solubilizers and sweeteners selected from the group consisting of One or more may be used.As a non-limiting example, the heavy agent (diluent) is mannitol (nmnnitol), calcium carbonate (calcium carbonate), calcium phosphate dibasic (calcium phosphate dibasic), calcium phosphate tribasic (calcium phosphate tribasic) , calcium sulfate, microcrystalline cellulose, microcrystalline silicified cellulose, powdered cellulose, dextrates, dextrose, fructose , lactitol, lactose anhydrous, lactose monohydrate, lactose dihydrate, lactose trihydrate, sorbitol, nmnnitol sorbitol, starch, starch At least one selected from the group consisting of pregelatinized starch, sucrose, talc, xylitol, maltose maltodextrin and maltitol; the disintegrant is crospovidone, alginic acid, Carbon dioxide, carboxymethyl cellulose calcium, carboxymethyl cellulose small, microcrystalline cellulose, powdered cellulose, large loscarmellose sodium, crospovidone, soium docusate, guar gum, hydroxypropyl cellulose, methylcellulose, polacrilin potassium, poloxamer, povidone, sodium alginate, sodium glycine carbonate, sodium lauryl sulfate, sodium at least one selected from the group consisting of starch glycolate, starch and pregelatinized starch; The binder is hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, acacia mucilage, alginic acid, carbomer, carboxymethyl cellulose calcium, carboxymethyl cellulose small, microcrystals Sex Cellulose, Powdered Cellulose, Ethyl Cellulose, Gelatin, Liquid Glucose, Guar Gum, Maltotextrin, Methyl Cellulose, 2021/250606 €1/162021/055105 at least one selected from the group consisting of polytextrose, polyethylene oxide, povidone, sodium alginate, starch paste, pregelatinized starch and sucrose; Lubricants (lubricants) include talc, sodium stearyl fumarate, magnesium stearate, colloidal silicon dioxide, polyethylene glycol 4000, polyethylene glycol 6000, sodium lauryl sulfate, starch,

At least one selected from the group consisting of hydrogenated castor oil, stearic acid, glyceryl palmitostearate, glyceryl monostearate, calcium silicate, powdered cellulose and starch may be used, but is not limited thereto. The pharmaceutical formulation for oral administration according to the present invention is preferably a capsule, pellet or tablet, but is not limited thereto, and the tablet is more preferably a film-coated tablet including a film coating layer. The film coating layer is selected from the group consisting of polyvinyl alcohol, a copolymer of polyvinyl alcohol and polyethylene, hydroxypropylmethylcellulose, hydroxypropyl cellulose, polyvinylpyrrolidone, methacrylic acid copolymer, polyethylene oxide, and xanthan gum. It may be formed using one or more coating bases, and thus the film coating layer is polyvinyl alcohol, a copolymer of polyvinyl alcohol and polyethylene, hydroxypropylmethylcellulose, hydroxypropylcellulose, polyvinylpyrrolidone, meta One or more film coating bases selected from the group consisting of acrylic acid copolymers, polyethylene oxide and xanthan gum, but are not limited thereto. The therapeutic composition according to the present invention or a pharmaceutical formulation comprising the same can be used for the treatment of coronavirus infection, in particular, coronavirus infection-19. Accordingly, the present invention relates to the use of a therapeutic composition according to the present invention or a pharmaceutical formulation comprising the same for the treatment of coronavirus infection, particularly coronavirus infection-19, and a pharmaceutical according to the present invention to a patient in need of the treatment. It relates to a method of treating coronavirus infection, in particular, coronavirus infection-19, characterized in that the formulation is administered. The invention also relates to the use of a composition of the invention for the manufacture of a medicament for the treatment of a coronavirus infection. 2021/250606 - €1/162021/055105 Brief Description of the Drawings Figure 1 shows the 8 show 1 - (that 0 \ 2 showing the inhibition rate and cytotoxicity of the compound of Formula II according to the present invention and the conventional drug) It is a drawing.

(Show) Compound of Formula II F) Chloroquine ( 110100[1^116)

(€)Remdesivir(1 1]1 ₍ 16 1·)

([>) Lopinavir Mae !·) FIG. 2 shows a visual evaluation method of hamster lung tissue. Figure 3 shows the improvement rate (%) of the gross pneumonia lesion of each group as a result of the animal experiment. Figure 4 shows the results of the gross analysis and histopathological analysis of the lung tissue for each group. 5 shows the results of histopathological analysis of lung tissues for each group. 6 shows the results of scoring the results of gross analysis and histopathological analysis of lung tissue for each group. 7 shows the lung tissue for each group.

Shows the results of confirming the change in the expression of the N gene of the virus. DETAILED DESCRIPTION OF THE INVENTION AND PREFERRED EMBODIMENTS Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In general, the nomenclature used herein is those well known and commonly used in the art. Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention in more detail, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited by these examples. Therefore, the practical scope of the present invention is It will be defined by the appended claims and their equivalents. Example 1. COVID-19 inhibitory effect of a compound having the structure of Formula I 1.1 Dose-response curve analysis method by immunofluorescence method 384 - 1.2x10 ⁴ Vero cells (ArCC CCL-81) per well were inoculated into a tissue culture plate. After 24 hours, two-fold serial dilutions in DMSO, chloroquine (Chloroquine) (Sigma-Aldrich C6628), remdesivir (MedChemExpress HY-104077) and lopinavir ( Lopinavir) (SelleckChem S1380) was treated with 4 types of compounds at the highest concentration of 50 nM in cells. ^{About 1 hour after each compound treatment, cells were infected with SARS-CoV-2} (National Pathogen Resource Bank, Korea Centers for Disease Control and Prevention) at a multiplicity of infection (MOI) of 0.0125 in the BSL3 facility and cultured at 37 for 24 hours. Thereafter, the cells were fixed with 4% paraformaldehyde (PFA), followed by permeabilization. Anti-SARS-CoV-2 Nucleocapsid (N) primary antibody (Sino

Biological Inc.), cells were stained with 488-conjugated goat anti-rabbit IgG secondary antibody (Molecular Probes) and Hoechst 33342 (Molecular Probes). Fluorescence expression was imaged using a large-capacity image analysis device, Operetta (Perkin Elmer). The acquired images were analyzed using an internal analysis program, Image Mining (IM) software. The total number of cells per well was calculated as the number of Hoechst-stained nuclei, and the number of infected cells was calculated as the number of cells expressing the nucleocapsid protein. The infectivity (infection ratio) was calculated as the number of cells/species cells expressing the nucleocapsid protein. Infectivity per well was defined as 0% of the mean infectivity of wells containing uninfected cells (mock) in the same plate, and 100% of the average infectivity of wells containing untreated infected cells (0.5% DMSO group) in the same plate. was normalized. Response curves and ICso values according to drug concentration were calculated using the formula Y = Bottom + (Top Bottom)/(l + (ICso/X) Hillslope) using XLFit 4 (IDBS) software. 2021/250606 - €1/162021/055105 derived. All 105 ₀ values were measured in two replicates, and the reliability of the assay was verified as a value of - - and the percent coefficient of variation (%（:\0). 1.2 Dose-response curve 1«:) analysis result of each of the four compounds of Formula II, chloroquine (0110100[111116), remdesivir (1 1]1 (16 1), and lopinavir acid 0 1 1·) As a result of analyzing the 8 show 11½-(:0\^2 inhibition rate (¾1曲½011) of Chloroquine (011!0 句, remdesivir (1 111£ 1·) and lopinavir mountain 0 1 1·) were respectively 11.59, 11.50 and 13.99, respectively. It was confirmed that it exhibits an exceptionally excellent therapeutic effect even at the concentration (see Table 1 and FIG. 1). Therefore, it can be confirmed that the compound of formula (I) or the compound of formula (III), which is a prodrug thereof, according to the present invention has a very high effect in the treatment of coronavirus infection-19 ((: 0\^ )-19.

【Table 1】 Results of measurement of 8show16-(:0\ᄌ-2inhibition rate and cytotoxicity of each compound)

Example 2: Confirmation of the therapeutic effect of the compound of Formula II on 8sho1犯-(:0\2 virus infection using hamsters)

2-1: Animal testing method An 8-week-old female hamster (Golden Syrian hamster, Orient Bio Co., Ltd.) was used, and the general welfare of animals was carried out according to the standard work guideline and test plan, and the Experimental Animal Welfare Act , Guide for the Care and Use of Laboratory Animals (by ILAR publication), the animal test was conducted, and the animal test in this example was reviewed by the Korea Institute of Biotechnology and Biotechnology IACUC (Institutional Animal Care and Use Committee) (approval number: KRIBB-AEC-20193) and IBC (Institutional Biosafety Committee) It passed the deliberation (approval number: KRIBB-IBC_ ² 0 ² 00 ² 1 ^{6 ).} For hamsters, 8 mice in each group were used for a total of 56 animals. For SARS-CoV-2 virus (National Pathogen Resource Bank, Korea Centers for Disease Control and Prevention) infection ^{, 100 ii L of 2.0xl0 5} PFU/mL virus was administered through the nasal passages. did As a positive control, remdesivir (VulcanChem, Cat No. VC1031201, CAS No. 1809249-37-3) was diluted with corn oil (Sigma-Aldrich, Cat No. C8267) and used. The compound of Formula II (CWM91) and remdesivir were administered intraperitoneally to the lower right side of the abdomen using a 3 mL syringe. The dose was calculated as 10 mL/kg according to the body weight of the experimental animal. As for the dose of the compound used in the experiment, the 50 mg/kg dose was set as the high-dose group, and the medium and low doses were set to 25 mg/kg and 12.5 mg/kg, respectively, by placing an azeotrope 2. In addition, the control group (NC, normal control) not administered with the test substance and excipient, the vehicle control group administered with physiological saline (VeC, vehicle control), the positive control group administered with remdesivir 5 mg/kg (PC, positive control) and SARS the -CoV-2 was set up inoculated intranasally ⁽² .0xl0 ⁵ PFU / mL) to the infection viral infection group (VC, virus control). In order to evaluate the antiviral efficacy of the compound against SARS-CoV-2, after nasal inoculation with SARS-CoV-2, Formula 11 inoculated with the test substance Formula II at 12.5 mg/kg, 25 mg/kg, 50 mg/kg -12.5 mpk, 25 mpk, and 50 mpk groups were set, respectively. Table 2 shows the groups and doses of the hamster experimental group _.

【Table 2】

2021/250606 ?€1/162021/055105

Infection of 8 shows 11½- (0\^2 of experimental animals was performed through the non-strong drip method, and the non-strong drip method was

No. 2.0><10 ⁵ under top anesthesia ! ⁷ !!/!!!! _It was carried out by inhalation by evenly dripping 100 必 of the virus dilution with ^ into both nasal passages. 2.0><10 ⁵ Mo /止 -1· was established through preliminary test results and bibliographic review conducted by the test research institute. is known as the route of infection most similar to respiratory infection through droplets that infect the human body, so the nasal drip method was set as the infection method. The dose of the positive control substance, remdesivir, is 5 based on the dose applied to the human body.

was set.

Twice a day (before the start of work and after the end of work), the animals were observed for death and moribund conditions. However, on the day of acquisition of animals and the day of autopsy, observations were made once a day (before the start of the work or after the end of the work). General symptoms including mortality, moribund, appearance and behavior changes were observed, and the date, time, and duration were recorded. The experiment was carried out from the day of infection (1 ^> positive) 0) to the fourth day after infection (1 ^> positive) 4). All animals subject to autopsy (1^104)

After anesthesia through inhalation, blood was collected from the posterior vena cava, and the presence or absence of external abnormalities and the presence of abnormalities in the chest cavity were visually observed. 2-2: Mortality and general symptoms of experimental animals As a result of the weight change test, no animals died during the test, and no general symptoms related to the administration of the test substance were observed in all test groups during the test period. During the test period, no weight loss was observed in the control group and excipient control group). 2021/250606 ?€1/162021/055105 virus

In , weight loss was observed on the 2nd and 4th days of infection (1>positive) 4), and weight loss similar to that of the virus-infected group was observed in the positive control group (!>(:)) and the test substance administration group. 2-3: Confirmation of gross pneumococcal lesion improvement rate The lungs of all animals subjected to autopsy (1>104) were visually evaluated and expressed as gross lung lesion improvement rate. The visual evaluation method is shown in FIG. 2 . As a result of the autopsy, as shown in FIG. 3 , no macroscopic abnormalities were observed in the control group (:) and the excipient control group (:). As a result of evaluating the gross pneumococcal lesion improvement rate (%) of the control group based on 100%, in the pneumococcal virus-infected group (\ ), hyperemia/congestion and edema of the lung margins (11 ¾ ) were observed in 60-70% of the lung area. It was observed, and the overall improvement rate of pneumococcal lesions for 5 lung lobes was observed to be 1%. In the positive control group (1), the improvement rate of gross pneumococcal lesions was 24.8%. The rate of improvement of gross pneumococcal lesions in the test substance-administered group was 12.511 1<;administered group, 251L1<;administrated group, 50 11 1<; It was observed in 41.3%, 43.9% and 48.9% of the administration group, respectively.

2-4: Histopathological confirmation The excised lung tissue was fixed in 10% neutral formalin for 7 days. The fixed lung tissue was stained with hematoxylin and eosin, and histopathological findings were observed. The scoring criteria for lung tissue damage are shown in Table 3.

【Table 3】

2021/250606 1^(:1^ 2021/055105

As a result, as shown in FIGS. 4 and 5 , no abnormal findings were observed in the lung tissue in the control group (NC) and the vehicle control group (VeC). In the virus-infected group (VC), viral pneumonia such as lickening of epithelial cells and inflammatory cell infiltration were observed in the lung tissue. The histopathological abnormalities observed in the virus-infected group were also observed in the positive control group (PC). As a result of observing histopathological findings in the test substance-administered group, the degree of abnormality observed in the virus-infected group was observed to be low. The score for evaluating the degree of lung tissue damage in each group is shown in FIG. 6 .

2-5: Confirmation of SARS-CoV-2 N (nucleocapsid) gene expression Some of the lung tissues removed from each group of hamsters were pulverized with a lysis solution, and the supernatant was extracted using an RNA extraction kit (RNeasy Mini Kit, Qiagen). RNA was extracted. 1 or g cDNA was synthesized using the i Script cDNA Synthesis Kit by quantifying the RNA. SARS-CoV-2 N gene-specific primer (Forward: 5, -TAA TCA GAC AAG GAA CTG ATT A-3, (SEQ ID NO: 1) // Reverse: 5, -CGA AGG TGT GAC TTC CAT G-3' ( SEQ ID NO: 2)) was analyzed in a CFX96 Touch Real-Time PCR Detection System (Bio-Rad). Real-time polymerase chain reaction on lung tissue through autopsy on the 4th day after infection

As a result of comparing the expression levels of N gene between the virus-infected group (VC) and the test group using the (real-time quantitative PCR) method, as shown in Fig. 7, the virus-infected group (VC) ) was expressed as 1, the positive control group (PC, Remdesivir 5 mpk) showed 0.47, whereas the expression levels of 0.27, 0.12 and 0.15 in the test substance administration group CWM91-12.5 mpk, CWM91-25 mpk and CWM91-50 mpk group, respectively. , it was confirmed that the expression of the SARS-CoV virus N gene was significantly reduced in the test substance administration group. INDUSTRIAL APPLICABILITY The compounds of Formulas I to VI according to the present invention efficiently inhibit SARS-Cov-2, thereby achieving an excellent effect in the treatment of COVID-19. «K For those of ordinary skill in the art who have described specific parts of the present invention in detail above, these specific descriptions are only preferred embodiments, and the scope of the present invention is not limited thereby. will be white Accordingly, it is intended that the substantial scope of the present invention be defined by the appended claims and their equivalents.

Claims

2021/250606 ?€1/162021/055105 Claims

^{[Claim 1] A composition for treating coronavirus infection-19 ( (} :0\ )-19) comprising a compound having the structure of Formula I, a prodrug thereof, a salt thereof, or an isomer thereof:

[Formula I]

In the formula I, it is ¾ (in: 1不₆ alkyl group, 02不₆ alkenyl group, or ₆ of the 02不

Alkyl group, an alkynyl group of 02不₆ alkenyl group, or 02不_6.

[Claim 2] The method according to claim 1, wherein in Formula I, ¾ is ⁽ : an alkyl group of 1不₆ _{or 02不6}

Coronavirus Infectious Disease- 1 ⁹ ((：0\^正)-1 ⁹ ) A composition for treatment.

[Claim 3] The composition for treating coronavirus infection-19 ((: 0\ )-19) according to claim 1, wherein the compound of Formula I has the structure of Formula II.

[Formula II] 2021/250606 1^(:1^2021/055105

[Claim 4] The prodrug of the compound having the structure of formula (I) according to claim 1, wherein the prodrug of the compound having the structure of formula (I) is used for treatment of coronavirus infection _ 19 ((:0\^ positive)-19) composition.

[Formula III]

In Formula III, ¾ is an alkyl group of 1不6, an alkenyl group of 02不6, or an alkynyl group of 02不6, and ¾ is an aryl group, a substituted aryl group, or -(:(=0)¾ and, at this time, is ¾ (1- (: an alkyl group of _6, 02不6 of the alkenyl group, or 02不6 alkynyl group, is ¾ ^{-1> 03¾ ^ 03 ^ + -} 1> 03 2明2 + , -1 ^> 03 ² ¾ ₂ ⁺ ,- 1>03 ² -1^句²⁺ , -1>03 ^{2 2+} , Structural Formula 1, Structural Formula 2, or Structural Formula 3.

[Claim 5] The compound according to claim 4, wherein in the compound of Formula III, ¾ is (: an alkyl group of 16) 2021/250606 ?€1/162021/055105 or C2-C6^\ alkenyl group, ¾ is -(：(=0)¾, ¾ is （：1不6 alkyl group, ¾ is -則知出, - HP03 Na ⁺ or [Coronavirus infection- 19 ( 3^/positive)- 19 characterized in that it is dark, a composition for treatment.

[Claim 6] The method according to claim 4, wherein in the compound of Formula III, ¾ is methyl ), and ¾ is - (the (=0) (: ¾, ¾ is -1>03¾, -1 03明 or [Coronavirus infection, characterized in that it is dark - ¹⁹ （（：0\^ ） - 1 ⁹ ) A composition for treatment.

[Claim 7] The coronavirus infection disease- according to claim 5, wherein the compound of Formula III has a structure selected from Formulas IV to VI.

1 ⁹ （（：0\^正）-1 ⁹ ) Therapeutic composition.

[Claim 8] The coronavirus infection according to any one of claims 1 to 7, which comprises at least one pharmaceutically acceptable carrier-

19 (COVID-19) therapeutic composition.

[Claim 9] A pharmaceutical formulation for oral or parenteral administration comprising the composition for treatment according to claim 8.

[Claim 10] The method of claim 9, wherein the parenteral administration is intranasal, intranasal, buccal, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, enteral, topical , A pharmaceutical formulation characterized in that it is administered sublingually or rectal.

[Claim 11] The pharmaceutical formulation according to claim 10, wherein the intranasal or intranasal administration is administered in the form of a spray, aerosol or inhalation.

[Claim 12] The pharmaceutical formulation according to claim 11, wherein the oral administration is administered in the form of a capsule, pellet, tablet, aqueous suspension or solution.