KR20210002386A - Novel quinolinone derivatives, preparation method thereof, and an antiviral composition containing the same as an active ingredient - Google Patents

Abstract

The present invention relates to a novel quinolinone derivative, a manufacturing method thereof, and an antiviral composition containing the same as an active component. The novel quinolinone derivative provided in an aspect of the present invention exhibits an excellent inhibitory activity on coronavirus infection, thereby being able to be usefully used as a pharmaceutical composition for preventing or treating diseases caused by coronavirus infection, that is, MERS.

Description

Novel quinolinone derivatives, preparation method thereof, and an antiviral composition containing the same as an active ingredient}

It relates to a novel quinolinone derivative, a method for preparing the same, and an antiviral composition containing the same as an active ingredient.

Coronavirus is a single-stranded positive RNA virus with an envelope, and its genome size is 25-32 kb, and it belongs to a relatively large virus among RNA viruses known so far. It is said to have a specific structure in the shape of a flame or crown because of the spike protein, which is a club-shaped protrusion, embedded in the shell, and the virus name is said to be derived from the Latin word Corona.

Coronaviruses found in various birds and animals such as bats, birds, cats, dogs, cows, pigs, and mice after being first discovered in chickens in 1937 are divided into four groups (Alpha-, Beta-, Gamma-, and Deltacoronavirus). Lose. The alpha and beta coronavirus groups are primarily infected with mammals, while the gamma and delta coronavirus groups can be found in birds. Coronavirus is known to cause various diseases such as gastrointestinal and respiratory diseases in animals.

Human coronaviruses that infect humans are HCoV-229E and HCoV-OC43, discovered in the 1960s, and HCoV-NL63 (2004) and HCoV-HKU1 (2005), discovered after the SARS pandemic.These are generally related to upper respiratory tract infections. Although known, it can also induce severe lung disease in immunodeficient patients. It is reported that the infection rate for coronavirus increases mainly in winter or early spring, and it is known that the proportion of coronavirus as a causative agent among adult cold patients is quite high. SARS-CoV, which causes Severe Acute Respiratory Syndrome (SARS), was first discovered in 2003, and according to the World Health Organization (WHO) report, it affected 8,273 patients worldwide from 2002 to 2003. There were 775 deaths (about 10% mortality), and additional cases and deaths were reported by 2004.

In September 2012, a patient with moderate respiratory disease with respiratory symptoms similar to SARS, such as high fever, cough, and shortness of breath, occurred, and the causative agent was identified as a novel coronavirus (HCoV-EMC), which is different from the known virus.

The novel coronavirus was classified under the name Middle East respiratory syndrome-coronavirus (MERS-CoV) by the Coronavirus Study Group of the International Committee on Taxonomy of Viruses in May 2013. This virus was presumed to be the most potent source of infection because its genetic sequencing is similar to Pipistrellus Bat CoV-HKU5 and HKU4 found in bats. However, in a recent paper published in the journal The Lancet Infectious Diseases, an analysis of blood samples from 50 dromedaries in Oman revealed that all traces of MERS-CoV antibodies were found. However, in the Canary Islands, 15 out of 105 were antibody-positive.

Although the virus itself has not been found, research has shown that these camels were infected with MERS or similar viruses at some point, and that the virus host was very likely to be camels.

Middle East Respiratory Syndrome Coronavirus (MERS-CoV) infection was officially reported to the WHO by June 2014 after the first case of infection was confirmed in Saudi Arabia in September 2012, and 808 patients worldwide and 313 died. (34.5%).

At the time of the initial confirmed case of Middle East Respiratory Syndrome Coronavirus (MERS-CoV), the clinical symptoms of the patient were quite similar to that of SARS, so there were concerns that it might have a significant association with SARS coronavirus, but after the full-length gene was analyzed, molecular genetics. It turned out to be a new virus that differs significantly from the SARS virus.

First of all, the Middle East Respiratory Syndrome Coronavirus (MERS-CoV) showed a low nucleotide homology of about 55% with the SARS coronavirus, and the two viruses also showed different evolutionary differences in the phylogenetic analysis. Middle East Respiratory Syndrome coronavirus (MERS-CoV) in group B of beta coronavirus belonged to group C of beta coronavirus. As belonging to the beta coronavirus group, SARS coronavirus and human coronaviruses HCoV-HKU1 and HCoV-OC43 are known.

In addition, SARS and the Middle East Respiratory Syndrome Coronavirus (MERS-CoV) also show a difference in the pathways that enter the cells.The SARS virus is the ACE2 receptor distributed in the ciliary cells of the human airways, and the Middle East Respiratory Syndrome Coronavirus (MERS-CoV) is DDP4 (also called CD26) is used in non-cilia cells. Both receptors are more distributed in the lower respiratory tract than in the upper respiratory tract of humans, and although the Middle East Respiratory Syndrome coronavirus (MERS-CoV) and SARS virus have high mortality rates, influenza or influenza must be transmitted through large droplets after progressing to pneumonia. Compared to aerosol-transmitted diseases, such as measles, the contagiousness may be lower. However, in the case of the outbreak of SARS, the infectious risk is still high because the transmission power can be considerably high in the case of close exposure to the SARS case or in an environment that causes the aerosolization of respiratory droplets.

What was the first source of infection of the Middle East Respiratory Syndrome Coronavirus (MERS-CoV), and how it spread to humans and caused the disease is not yet known. However, two cases of confirmed cases of the novel coronavirus that occurred were found to have visited the farm and contacted animals. This shows the possibility of a common transmission of the novel coronavirus.

The main clinical symptoms are fever (87%), cough (89%), and shortness of breath. Some patients also show vomiting and diarrhea (35%). Renal failure also occurs in patients with reduced immune function, and the mortality rate is very high at 34.5%. It occurs a lot in the Middle East (Saudi Arabia, Qatar, etc.), and these regions are estimated to be infected areas, but the exact route of infection has not been accurately identified. However, there has been no evidence of widespread infection between humans, but it has been confirmed that it is spread through close contact with family members or medical staff. The incubation period is estimated to be 9 to 12 days, but there are many differences in incubation period for each person.

Until now, drugs and preventive vaccines that have confirmed the therapeutic effect of the Middle East Respiratory Syndrome Coronavirus (MERS-CoV) have not been developed. Therefore, until now, there are no special treatment and prevention methods other than avoiding contact with patients suspected of viral infection and thorough personal hygiene. Currently, antibiotics, ribavirin, interferon, and patient serum have been used to treat infections of the Middle East Respiratory Syndrome coronavirus (MERSCoV), but these treatments have not exhibited specific effects. However, recently, it has been known that radotinib has the potential to treat respiratory viral diseases (Patent Document 1).

Therefore, the development of alternative drugs for the treatment and prevention of diseases for the Middle East Respiratory Syndrome Coronavirus (MERS-CoV) is a very urgent situation.

Republic of Korea Patent Publication 10-2017-0009276

An object of the present invention is to provide a novel quinolinone derivative, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof that can be used as a therapeutic agent for Middle East Respiratory Syndrome coronavirus infection.

Another object of the present invention is to provide a method for preparing the novel quinolinone derivative.

Another object of the present invention is to provide a pharmaceutical composition for antiviral containing the novel quinolinone derivative, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of MERS containing the novel quinolinone derivative, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient. Is to do.

Another object of the present invention is to prevent or improve the health of diseases caused by viral infections containing the novel quinolinone derivatives, solvates thereof, hydrates thereof, or pharmaceutically acceptable salts thereof as an active ingredient. It is to provide a nutraceutical composition.

Another object of the present invention is a virus comprising administering the novel quinolinone derivative, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof to a subject in need thereof. It is to provide a method of treating diseases caused by infection.

Another object of the present invention is to provide the novel quinolinone derivative, pharmaceutical composition, and/or health functional food composition for use in the prevention or treatment of diseases caused by viral infection.

Another object of the present invention is to provide a use of the novel quinolinone derivative for preparing a medicament for use in the prevention or treatment of diseases caused by viral infection.

To achieve the above object,

One aspect of the present invention provides a compound represented by Formula 1 below, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.

[Formula 1]

(In Formula 1,

R ¹ is independently -H, -OH, halo, -NO ₂ , -NH ₂ , -CN, C _1-10 straight or branched chain alkyl unsubstituted or substituted with one or more halogens, or unsubstituted or one or more Halogen-substituted C _1-10 linear or branched alkoxy;

R ² is unsubstituted or C _1-10 straight or branched chain alkyl substituted with one or more halogens, or C _3-10 unsubstituted or substituted C _3-10 cycloalkyl;

R ³ is independently -H, -OH, halo, -NO ₂ , -NH ₂ , -CN, straight or branched C _1-10 alkyl unsubstituted or substituted with one or more halogens, or unsubstituted or one or more Halogen-substituted C _1-10 linear or branched alkoxy;

n is an integer from 0 to 4; And

m is an integer from 0 to 5).

In addition, one aspect of the present invention, as shown in the following Scheme 1,

Preparing a compound represented by Formula 3 from the compound represented by Formula 2;

Preparing a compound represented by Formula 4 from the compound represented by Formula 3 prepared in the above step;

Preparing a compound represented by Formula 5 from the compound represented by Formula 4 prepared in the above step;

Preparing a compound represented by Formula 6 from the compound represented by Formula 5 prepared in the above step; And

Preparing a compound represented by Formula 1 from the compound represented by Formula 6 prepared in the above step; It provides a method for preparing the compound represented by Formula 1, including.

[Scheme 1]

(In the above Scheme 1,

R ¹ , R ² , R ³ , n and m are independently as defined in Formula 1).

Further, an aspect of the present invention provides an antiviral pharmaceutical composition containing the compound represented by Formula 1, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, an aspect of the present invention provides a pharmaceutical composition for preventing or treating MERS containing the compound represented by Formula 1, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient. .

Further, an aspect of the present invention is a health functional food for preventing or improving diseases caused by viral infection containing the compound represented by Formula 1, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient. The composition is provided.

In addition, an aspect of the present invention is a viral infection comprising administering a compound represented by Formula 1, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof to a subject in need thereof. Provides treatment methods for diseases caused by.

Further, an aspect of the present invention provides a compound represented by Formula 1, a pharmaceutical composition, and/or a health functional food composition for use in the prevention or treatment of diseases caused by viral infection.

In addition, an aspect of the present invention provides a use of the compound represented by Formula 1 for preparing a medicament for use in the prevention or treatment of diseases caused by viral infection.

Since the novel quinolinone derivative provided in one aspect of the present invention exhibits excellent inhibitory activity against coronavirus infection,

It can be usefully used as a pharmaceutical composition for preventing or treating diseases caused by coronavirus infection, that is, MERS.

Hereinafter, the present invention will be described in detail.

One aspect of the present invention provides a compound represented by Formula 1 below, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.

[Formula 1]

(In Formula 1,

R ¹ is independently -H, -OH, halo, -NO ₂ , -NH ₂ , -CN, C _1-10 straight or branched chain alkyl unsubstituted or substituted with one or more halogens, or unsubstituted or one or more Halogen-substituted C _1-10 linear or branched alkoxy;

R ² is unsubstituted or C _1-10 straight or branched chain alkyl substituted with one or more halogens, or C _3-10 unsubstituted or substituted C _3-10 cycloalkyl;

R ³ is independently -H, -OH, halo, -NO ₂ , -NH ₂ , -CN, straight or branched C _1-10 alkyl unsubstituted or substituted with one or more halogens, or unsubstituted or one or more Halogen-substituted C _1-10 linear or branched alkoxy;

n is an integer from 0 to 4; And

m is an integer from 0 to 5).

On the other side,

R ¹ is independently -H, -OH, halo, -NO ₂ , -NH ₂ , -CN, C _1-5 straight or branched chain alkyl unsubstituted or substituted with one or more halogens, or unsubstituted or one or more Halogen-substituted C _1-5 straight or branched alkoxy;

R ² is methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, tert-butyl, or cyclopropyl;

R ³ is independently -H, -OH, halo, -NO ₂ , -NH ₂ , -CN, straight or branched C _1-5 alkyl unsubstituted or substituted with one or more halogens, or unsubstituted or one or more Halogen-substituted C _1-5 straight or branched alkoxy;

n is an integer from 0 to 4; And

m may be an integer from 0 to 4.

On the other side,

R ¹ is independently -H, -F, -Cl, -Br, -NO ₂ , -CF ₃ , methyl or ethyl;

R ² is methyl, ethyl, or iso-propyl;

R ³ is independently -H, -F, -Cl, -Br, -CF ₃ , methyl, or methoxy;

n is an integer from 0 to 3; And

m may be an integer of 0 to 3.

On the other side,

The compound represented by Formula 1 may be any one compound selected from the following compound group.

(1) 3-acetyl-8-chloro-5-methyl-2-(2,3,4-trifluorophenylamino)quinolin-4(1H)-one;

(2) 3-acetyl-6,8-difluoro-2-(2,3,4-trifluorophenylamino)quinolin-4(1H)-one;

(3) 3-acetyl-6-chloro-2-(3,5-dichlorophenylamino)-8-nitroquinolin-4(1H)-one;

(4) 3-acetyl-5,6,8-trichloro-2-(3-fluorophenylamino)quinolin-4(1H)-one;

(5) 6,8-difluoro-3-isobutyryl-2-(2,3,4-trifluorophenylamino)quinolin-4(1H)-one;

(6) 5,8-dichloro-2-(2,4-dichlorophenylamino)-3-isobutyrylquinolin-4(1H)-one;

(7) 5,8-dichloro-3-isobutyryl-2-(2,3,4-trifluorophenylamino)quinolin-4(1H)-one;

(8) 5,8-dichloro-3-isobutyryl-2-(4-methoxyphenylamino)quinolin-4(1H)-one;

(9) 3-acetyl-2-(2,4-difluorophenylamino)-6,8-difluoroquinolin-4(1H)-one;

(10) 3-acetyl-2-(2,4-dichlorophenylamino)-6,8-difluoroquinolin-4(1H)-one;

(11) 3-acetyl-6-chloro-2-(2,4-dichlorophenylamino)-8-nitroquinolin-4(1H)-one;

(12) 3-acetyl-5,8-dichloro-2-(2,3,4-trifluorophenylamino)quinolin-4(1H)-one;

(13) 5,8-dichloro-2-(3,5-difluorophenylamino)-3-isobutyrylquinolin-4(1H)-one;

(14) 2-(2,4-difluorophenylamino)-6,8-difluoro-3-isobutyrylquinolin-4(1H)-one;

(15) 5,8-dichloro-2-(4-chloro-2-fluorophenylamino)-3-isobutyrylquinolin-4(1H)-one;

(16) 3-acetyl-5,8-dichloro-2-(2,4-dichlorophenylamino)quinolin-4(1H)-one;

(17) 3-acetyl-5,8-dichloro-2-(2,4-difluorophenylamino)quinolin-4(1H)-one;

(18) 3-acetyl-8-chloro-5-(trifluoromethyl)-2-(2,3,4-trifluorophenylamino)quinolin-4(1H)-one;

(19) 3-acetyl-8-chloro-2-(2,4-dichlorophenylamino)-5-(trifluoromethyl)quinolin-4(1H)-one;

(20) 3-acetyl-2-(2,4-dichlorophenylamino)-5,8-difluoroquinolin-4(1H)-one;

(21) 3-acetyl-2-(4-chlorophenylamino)-6,8-difluoroquinolin-4(1H)-one;

(22) 3-acetyl-2-(2,4-dichlorophenylamino)-8-ethylquinolin-4(1H)-one;

(23) 3-acetyl-2-(2,5-difluorophenylamino)-5,8-difluoroquinolin-4(1H)-one;

(24) 3-acetyl-6-chloro-2-(3-chlorophenylamino)-8-nitroquinolin-4(1H)-one;

(25) 3-acetyl-5,8-dichloro-2-(4-chloro-2-fluorophenylamino)quinolin-4(1H)-one;

(26) 3-acetyl-5,8-dichloro-2-(4-chloro-2-(trifluoromethyl)phenylamino)quinolin-4(1H)-one;

(27) 5,8-dichloro-2-(2,5-difluorophenylamino)-3-propionylquinolin-4(1H)-one;

(28) 3-acetyl-6-chloro-2-(4-fluorophenylamino)-8-nitroquinolin-4(1H)-one;

(29) 3-acetyl-6-chloro-2-(2-chloro-4-fluorophenylamino)-8-nitroquinolin-4(1H)-one;

(30) 3-acetyl-8-chloro-2-(2,3-difluorophenylamino)-5-nitroquinolin-4(1H)-one;

(31) 3-acetyl-2-(3,5-difluorophenylamino)-7,8-difluoroquinolin-4(1H)-one;

(32) 3-acetyl-2-(4-bromophenylamino)-8-chloro-5-fluoroquinolin-4(1H)-one;

(33) 3-acetyl-2-(4-chloro-2-fluorophenylamino)-5,8-difluoroquinolin-4(1H)-one;

(34) 3-acetyl-2-(3,5-difluorophenylamino)-7,8-difluoroquinolin-4(1H)-one;

(35) 3-acetyl-2-(2-chloro-4-methylphenylamino)-5,8-difluoroquinolin-4(1H)-one;

(36) 3-acetyl-7,8-dichloro-2-(3,5-difluorophenylamino)quinolin-4(1H)-one;

(37) 3-acetyl-8-chloro-5-fluoro-2-(2-fluoro-5-methylphenylamino)quinolin-4(1H)-one;

(38) 5,8-dichloro-2-(2,4-difluorophenylamino)-3-propionylquinolin-4(1H)-one;

(39) 3-acetyl-2-(4-bromophenylamino)-6,8-difluoroquinolin-4(1H)-one;

(40) 3-acetyl-6-chloro-2-(2,4-dichlorophenylamino)-8-nitroquinolin-4(1H)-one;

(41) 5,8-dichloro-2-(3,5-difluorophenylamino)-3-isobutyrylquinolin-4(1H)-one;

(42) 3-acetyl-2-(4-bromophenylamino)-7,8-dichloroquinolin-4(1H)-one;

(43) 3-acetyl-5,8-dibromo-2-(3,5-difluorophenylamino)quinolin-4(1H)-one; And

(44) 3-acetyl-5,8-difluoro-2-(4-(trifluoromethyl)phenylamino)quinolin-4(1H)-one.

The compound represented by Formula 1 of the present invention can be used in the form of a pharmaceutically acceptable salt, and an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid, etc., aliphatic mono and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanes. Non-toxic organic acids such as dioates, aromatic acids, aliphatic and aromatic sulfonic acids, trifluoroacetic acid, acetate, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid, etc. Get from Examples of such pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, i. Odide, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, sube Rate, sebacate, fumarate, maleate, butine-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitro benzoate, hydroxybenzoate, Methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, Glycolate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate, and the like.

The acid addition salt according to the present invention can be prepared by a conventional method, for example, a precipitate formed by dissolving the derivative of Formula 1 in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile, etc. and adding an organic or inorganic acid May be prepared by filtration and drying, or may be prepared by distilling off a solvent and an excess of acid under reduced pressure and then drying to crystallize under an organic solvent.

In addition, a pharmaceutically acceptable metal salt can be made using a base. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and evaporating and drying the filtrate. At this time, it is pharmaceutically suitable to prepare sodium, potassium or calcium salt as the metal salt. In addition, the corresponding salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable negative salt (eg, silver nitrate).

Further, the present invention includes not only the compound represented by Formula 1 and its pharmaceutically acceptable salts, but also solvates, optical isomers, hydrates, etc. that may be prepared therefrom.

Another aspect of the present invention is as shown in Scheme 1 below,

Preparing a compound represented by Formula 3 from the compound represented by Formula 2;

Preparing a compound represented by Formula 4 from the compound represented by Formula 3 prepared in the above step;

Preparing a compound represented by Formula 5 from the compound represented by Formula 4 prepared in the above step;

Preparing a compound represented by Formula 6 from the compound represented by Formula 5 prepared in the above step; And

Preparing a compound represented by Formula 1 from the compound represented by Formula 6 prepared in the above step; It provides a method for preparing the compound represented by Formula 1, including.

[Scheme 1]

(In the above Scheme 1,

R ¹ , R ² , R ³ , n and m are independently as defined in Formula 1).

Hereinafter, a method for preparing the compound represented by Formula 1 provided in one aspect of the present invention will be described in detail step by step.

In the method for preparing the compound represented by Formula 1 provided in an aspect of the present invention, step 1 is a step of preparing a compound represented by Formula 3 from the compound represented by Formula 2. More specifically, by reacting a diketene derivative with an aniline derivative represented by Chemical Formula 2, an amide derivative represented by Chemical Formula 3 is prepared. Benzene or the like may be used as the reaction solvent, and the reaction temperature may be in the range of 80 to 140°C, and the reaction time may be performed for 70 to 140 minutes, but is not particularly limited thereto.

In the method for preparing the compound represented by Formula 1 provided in an aspect of the present invention, step 2 is a step of preparing a compound represented by Formula 4 from the compound represented by Formula 3 prepared in the step. More specifically, dimethyl sulfate and CS ₂ are reacted with the amide derivative represented by Formula 3 to prepare a compound represented by Formula 4. DMF or the like may be used as the reaction solvent, the reaction temperature may be in the range of room temperature (about 20 to 25°C), and the reaction time may be performed for 30 to 200 minutes, but is not limited thereto.

In the method for preparing the compound represented by Formula 1 provided in an aspect of the present invention, step 3 is a step of preparing a compound represented by Formula 5 from the compound represented by Formula 4 prepared in the step. More specifically, a bis(methylthio) compound represented by Formula 4 is dissolved in o-dichlorobenzene and stirred to prepare a compound represented by Formula 5. At this time, the stirring temperature may be 160 to 200°C, and the stirring time may be performed for 120 minutes to 360 minutes, but is not particularly limited thereto.

In the method for preparing the compound represented by Formula 1 provided in an aspect of the present invention, step 4 is a step of preparing a compound represented by Formula 6 from the compound represented by Formula 5 prepared in the step. More specifically, the quinolin-4-one compound represented by Chemical Formula 5 is dissolved in acetic acid, and hydrogen peroxide solution is added and stirred to prepare a compound represented by Chemical Formula 6. At this time, the stirring temperature may be 30 to 70° C., and the stirring time may be performed for 120 to 240 minutes, but is not particularly limited thereto.

In the method for preparing the compound represented by Formula 1 provided in an aspect of the present invention, step 5 is a step of preparing a compound represented by Formula 1 from the compound represented by Formula 6 prepared in the step. More specifically, a sulfoxide compound represented by Formula 6 is dissolved in diphenyl ether, and an aniline derivative is added to prepare a compound represented by Formula 1. At this time, the reaction temperature may be 160 to 200°C, and the reaction time may be performed for 60 to 180 minutes, but is not particularly limited thereto.

Another aspect of the present invention provides a pharmaceutical composition for antiviral containing a compound represented by Formula 1, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

The antiviral pharmaceutical composition includes corona virus, influenza virus, Alpui virus, ring virus, bovine diarrhea virus, chikungunya virus, dengue virus, hepatitis B virus, hepatitis C virus, human cytomegalovirus, human immunodeficiency Virus, Ilheus virus, Japanese encephalitis virus, Cocovera virus, Kunjin virus, Kiasanur forest disease virus, Leap disease virus, Measles virus, Metapneumovirus, Mosaic virus, Murray Valley virus, Parainfluenza virus, Polio virus, Powasan Virus, respiratory syncytial virus, Roshio virus, St. Louis encephalitis virus, tick-mediated encephalitis virus, West Nile virus, and can exhibit antiviral activity against one or more selected from the group consisting of yellow fever virus.

On the other hand, the coronavirus is infectious bronchitis virus, bovine coronavirus, canine coronavirus, feline coronavirus, porcine erythrocyte agglutinating encephalomyelitis virus, mouse hepatitis virus, turkey coronavirus, infectious gastrointestinal disease virus, rat coronavirus, human coronavirus 229E (HCoV -229E), one selected from the group consisting of human coronavirus NL63 (HCoV-NL63), human coronavirus OC43 (HCoV-OC43), SARS coronavirus (SARS-CoV) and Middle East Respiratory Syndrome coronavirus (MERS-Cov) It can be more than that. The antiviral pharmaceutical composition may be used for the prevention or treatment of diseases caused by infection with Middle East Respiratory Syndrome coronavirus (MERS-Cov), for example MERS.

Another aspect of the present invention provides a pharmaceutical composition for preventing or treating MERS containing a compound represented by Formula 1, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

The compound represented by Formula 1 or a pharmaceutically acceptable salt thereof may be administered in various oral and parenteral formulations upon clinical administration. In the case of formulation, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants that are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and these solid preparations include at least one excipient in one or more compounds, such as starch, calcium carbonate, sucrose, or lactose ( lactose), gelatin, etc. In addition, in addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, liquid solutions, emulsions, and syrups.In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as humectants, sweeteners, fragrances, and preservatives may be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, and emulsions. As the non-aqueous solvent and suspension, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used.

A pharmaceutical composition containing the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient can be administered parenterally, and parenteral administration is by subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection It depends on how to do it.

At this time, in order to formulate a formulation for parenteral administration, the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof is mixed in water together with a stabilizer or buffer to prepare a solution or suspension, and the ampoule or vial unit dosage form It can be manufactured with The composition may be sterilized and/or contain adjuvants such as preservatives, stabilizers, hydrating agents or emulsification accelerators, salts and/or buffers for controlling osmotic pressure, and other therapeutically useful substances, which are conventional methods of mixing, granulation. It can be formulated according to the method of painting or coating.

Formulations for oral administration include, for example, tablets, pills, hard/soft capsules, solutions, suspensions, emulsifiers, syrups, granules, elixirs, and troches.These formulations include diluents (e.g., lactose) in addition to the active ingredients. , Dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine), lubricants (e.g. silica, talc, stearic acid and its magnesium or calcium salt and/or polyethylene glycol). Tablets may contain a binder such as magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidine, and in some cases, boron such as starch, agar, alginic acid or sodium salt thereof. It may contain release or boiling mixtures and/or absorbents, colorants, flavoring agents, and sweetening agents.

Another aspect of the present invention is a health functional food composition for the prevention or improvement of diseases caused by viral infection containing the compound represented by Formula 1, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient. to provide. At this time, the disease caused by the viral infection may be MERS.

The compound represented by Formula 1 according to the present invention may be added to food as it is or may be used with other foods or food ingredients, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined depending on the purpose of use (for prevention or improvement). In general, the amount of the compound in the health food may be added in 0.1 to 90 parts by weight of the total food weight. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of health control, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

In addition, the health functional beverage composition of the present invention is not particularly limited to other ingredients other than containing the compound as an essential ingredient in the indicated ratio, and may contain various flavoring agents or natural carbohydrates, etc. as additional ingredients, as in ordinary beverages. have. Examples of the natural carbohydrates described above include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose, and the like; And polysaccharides, for example, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (taumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 g of the composition of the present invention.

Further, in addition to the above, the compound represented by Formula 1 according to the present invention is a variety of nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavors and natural flavoring agents, coloring agents and heavy weight agents (cheese, chocolate, etc.), pect Acids and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonates used in carbonated beverages, and the like. In addition, the compound represented by Chemical Formula 1 of the present invention may contain flesh for the manufacture of natural fruit juice and fruit juice beverages and vegetable beverages.

Another aspect of the present invention is a viral infection comprising administering a compound represented by Formula 1, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof to a subject in need thereof. Provides treatment methods for diseases caused by.

In addition, another aspect of the present invention provides a compound represented by Formula 1, a pharmaceutical composition, and/or a functional food composition for use in the prevention or treatment of diseases caused by viral infection.

Further, another aspect of the present invention provides a use of the compound represented by Formula 1 for preparing a medicament for use in the prevention or treatment of diseases caused by viral infection.

Since the novel quinolinone derivative provided in one aspect of the present invention exhibits excellent inhibitory activity against coronavirus infection,

It can be usefully used as a pharmaceutical composition for the prevention or treatment of diseases caused by coronavirus infection, that is, MERS, which is supported by the Examples and Experimental Examples described below.

Hereinafter, the present invention will be described in detail through examples and experimental examples.

However, the examples and experimental examples described below are only to specifically illustrate the present invention in one aspect, and the present invention is not limited thereto.

<Example 1> 3-acetyl-8-chloro-5-methyl-2-(2,3,4-trifluorophenylamino)quinolin-4(1H)-one

The compound of Example 1 was obtained from the Korea Compound Bank.

(ChemBank#: 229924)

<Example 2> 3-acetyl-6,8-difluoro-2-(2,3,4-trifluorophenylamino)quinolin-4(1H)-one

The compound of Example 2 was obtained from the Korea Compound Bank.

(ChemBank#: 3892)

<Example 3> 3-acetyl-6-chloro-2-(3,5-dichlorophenylamino)-8-nitroquinolin-4(1H)-one

The compound of Example 3 was obtained from the Korea Compound Bank.

(ChemBank#: 230122)

<Example 4> 3-acetyl-5,6,8-trichloro-2-(3-fluorophenylamino)quinolin-4(1H)-one

The compound of Example 4 was obtained from the Korea Compound Bank.

(ChemBank#: 230020)

<Example 5> 6,8-difluoro-3-isobutyryl-2-(2,3,4-trifluorophenylamino)quinolin-4(1H)-one

Step 1: N-(2,4-difluorophenyl)-4-methyl-3-oxopentanamide

Method 1: Aniline (11.0 mmol) was dissolved in benzene (20 mL), diketene (10.0 mmol) and triethylamine (1.0 mmol) were added, followed by heating at 110° C. for 90 to 120 minutes. After the reaction was completed, the solution was removed under reduced pressure, and the residue was subjected to silica gel chromatography (developing solution ethyl acetate: hexane = 1: 2).

Method 2: Substituted acetyl acetate (10 mmol) and aniline (10 mmol) were added to xylene (20 mL) and heated at 120° C. for 2 hours. After the reaction was completed, the solution was removed under reduced pressure, and the residue was subjected to silica gel chromatography (developing solution ethyl acetate: hexane = 1: 2).

Step 2: 2-(bis(methylthio)methylene)-N-(2,4-difluorophenyl)-4-methyl-3-oxopentanamide

Dissolve the amide (17.5 mmol) prepared in step 1 in DMF (50 mL), add K ₂ CO ₃ (5.25 mmol) and tetrabutylammonium bromide (1.8 mmol), at room temperature (about 20-23°C) It stirred for 30 minutes. CS ₂ (21 mmol) was placed and stirred for 2 hours. Dimethyl sulfate (38.5 mmol) was added and stirred for 2 hours, and the reaction was put in ice water and stirred. The resulting solid was filtered, washed with hexane and dried.

Step 3: 6,8-difluoro-3-isobutyryl-2-(methylthio)quinolin-4(1H)-one

The bis(methylthio) compound (5 mmol) prepared in step 2 was dissolved in o-dichlorobenzene (10 mL), and stirred at 180° C. for 4 hours. The resulting solid was filtered, washed with hexane, and dried.

Step 4: 6,8-difluoro-3-isobutyryl-2-(methylsulfinyl)quinolin-4(1H)-one

The quinolin-4-one (2.6 mmol) prepared in step 3 was dissolved in 15 mL of acetic acid, and hydrogen peroxide (30%, 1.5 mL) was added, followed by stirring at 50° C. for 3 hours. The reaction solution was poured into ice water and stirred for 30 minutes. The resulting solid was filtered, washed with hexane and dried.

Step 5: 6,8-difluoro-3-isobutyryl-2-(2,3,4-trifluorophenylamino)quinolin-4(1H)-one

The compound (0.57 mmol) prepared in step 4 was dissolved in 4 mL of diphenyl ether, and aniline (0.62 mmol) was added. After reacting at 180° C. for 2 hours, column chromatography was performed.

1H NMR (300 MHz, CDCl ₃ ) δ 13.45 (s, 1H), 7.80 (dt, J = 8.8, 2.2 Hz, 1H), 7.62 (s, 1H), 7.24-7.15 (m, 2H), 7.10 (ddd , J = 10.3, 7.8, 2.8 Hz, 1H), 4.34 (hept, J = 6.6 Hz, 1H), 1.21 (d, J = 6.7 Hz, 6H).

<Example 6> 5,8-dichloro-2-(2,4-dichlorophenylamino)-3-isobutyrylquinolin-4(1H)-one

*

The title compound was prepared in a similar manner to Example 5 above.

1H NMR (300 MHz, CDCl ₃ ) δ 13.21 (s, 1H), 8.16 (s, 1H), 7.68-7.62 (m, 1H), 7.46-7.38 (m, 3H), 7.22 (d, J = 8.5 Hz , 1H), 4.21 (p, J = 6.7 Hz, 1H), 1.22 (d, J = 6.7 Hz, 6H).

<Example 7> 5,8-dichloro-3-isobutyryl-2-(2,3,4-trifluorophenylamino)quinolin-4(1H)-one

The title compound was prepared in a similar manner to Example 5 above.

1H NMR (300 MHz, CDCl ₃ ) δ 13.22 (s, 1H), 8.14 (s, 1H), 7.43 (d, J = 8.5 Hz, 1H), 7.22 (dd, J = 13.3, 8.1 Hz, 3H), 4.22 (tt, J = 13.3, 6.8 Hz, 1H), 1.22 (d, J = 6.7 Hz, 6H).

<Example 8> 5,8-dichloro-3-isobutyryl-2-(4-methoxyphenylamino)quinolin-4(1H)-one

The title compound was prepared in a similar manner to Example 5 above.

1H NMR (300 MHz, CDCl ₃ ) δ 12.83 (s, 1H), 8.31 (s, 1H), 7.36 (d, J = 8.5 Hz, 1H), 7.32-7.26 (m, 2H), 7.17 (d, J = 8.5 Hz, 1H), 7.10-6.99 (m, 2H), 4.21 (p, J = 6.7 Hz, 1H), 3.88 (s, 3H), 1.21 (d, J = 6.7 Hz, 6H).

<Example 9> 3-acetyl-2-(2,4-difluorophenylamino)-6,8-difluoroquinolin-4(1H)-one

The compound of Example 9 was obtained from the Korea Compound Bank.

(ChemBank#: 203744)

<Example 10> 3-acetyl-2-(2,4-dichlorophenylamino)-6,8-difluoroquinolin-4(1H)-one

The title compound was prepared in a similar manner to Example 5 above.

1H NMR (300 MHz, CDCl ₃ ) δ 13.38 (s, 1H), 7.79 (dt, J = 9.0, 3.0 Hz, 1H), 7.66 (d, J = 1.8 Hz, 1H), 7.63 (brs, 1H), 7.44 (d, J=2.1 Hz, 1H). 7.43 (s, 1H), 7.13-7.06 (m, 1H) 2.85 (s, 1H).

<Example 11> 3-acetyl-6-chloro-2-(2,4-dichlorophenylamino)-8-nitroquinolin-4(1H)-one

The compound of Example 11 was obtained from the Korea Compound Bank.

(ChemBank#: 3630)

<Example 12> 3-acetyl-5,8-dichloro-2-(2,3,4-trifluorophenylamino)quinolin-4(1H)-one

The title compound was prepared in a similar manner to Example 5 above.

1NMR (300 MHz, DMSO-d6) δ 12.55 (s, 1H), 7.69 (d, J = 9.0 Hz, 1H), 7.39-7.48 (q, J = 9.0, 18.0 Hz, 1H), 7.25 (d, J = 9.0 Hz, 1H), 2.59 (s, 3H).

<Example 13> 5,8-dichloro-2-(3,5-difluorophenylamino)-3-isobutyrylquinolin-4(1H)-one

The compound of Example 13 was obtained from the Korea Compound Bank.

(ChemBank#: 3916)

<Example 14> 2-(2,4-difluorophenylamino)-6,8-difluoro-3-isobutyrylquinolin-4(1H)-one

The title compound was prepared in a similar manner to Example 5 above.

1H NMR (300 MHz, CDCl ₃ ) δ 13.24 (s, 1H), 7.79 (dt, J = 8.7, 1.8 Hz), 7.59 (brs, 1H), 7.46-7.39 (m, 1H), 7.14-7.05 (m , 3H), 4.39-4.29 (m, 1H), 1.57 (s, 1H), 1.21 (d, J= 6.9 Hz, 1H).

<Example 15> 5,8-dichloro-2-(4-chloro-2-fluorophenylamino)-3-isobutyrylquinolin-4(1H)-one

The title compound was prepared in a similar manner to Example 5 above.

1H NMR (300 MHz, CDCl ₃ ) δ 13.16 (s, 1H), 8.17 (s, 1H), 7.42 (d, J = 8.5 Hz, 1H), 7.40-7.31 (m, 2H), 7.23 (d, J = 8.5 Hz, 2H), 4.20 (p, J = 6.7 Hz, 1H), 1.22 (d, J = 6.7 Hz, 6H).

<Example 16> 3-acetyl-5,8-dichloro-2-(2,4-dichlorophenylamino)quinolin-4(1H)-one

The compound of Example 16 was obtained from the Korea Compound Bank.

(ChemBank#: 2307)

<Example 17> 3-acetyl-5,8-dichloro-2-(2,4-difluorophenylamino)quinolin-4(1H)-one

The compound of Example 17 was obtained from the Korea Compound Bank.

(ChemBank#: 229646)

<Example 18> 3-acetyl-8-chloro-5-(trifluoromethyl)-2-(2,3,4-trifluorophenylamino)quinolin-4(1H)-one

The title compound was prepared in a similar manner to Example 5 above.

1H NMR (300 MHz, CDCl ₃ ) δ 13.10 (s, 1H), 8.24 (s, 1H), 7.67 (s, 2H), 7.22-7.28 (m, 2H), 2.82 (s, 3H).

<Example 19> 3-acetyl-8-chloro-2-(2,4-dichlorophenylamino)-5-(trifluoromethyl)quinolin-4(1H)-one

The title compound was prepared in a similar manner to Example 5 above.

1H NMR (300 MHz, CDCl ₃ ) δ 13.14 (s, 1H), 8.24 (s, 1H), 7.66 (s, 1H), 7.63 (s, 2H), 7.46 (s, 2H), 2.79 (s, 3H ).

<Example 20> 3-acetyl-2-(2,4-dichlorophenylamino)-5,8-difluoroquinolin-4(1H)-one

*

The compound of Example 20 was obtained from the Korea Compound Bank.

(ChemBank#: 3580)

<Example 21> 3-acetyl-2-(4-chlorophenylamino)-6,8-difluoroquinolin-4(1H)-one

The compound of Example 21 was obtained from the Korea Compound Bank.

(ChemBank#: 3893)

<Example 22> 3-acetyl-2-(2,4-dichlorophenylamino)-8-ethylquinoline-4(1H)-one

The compound of Example 22 was obtained from the Korea Compound Bank.

(ChemBank#: 4636)

<Example 23> 3-acetyl-2-(2,5-difluorophenylamino)-5,8-difluoroquinolin-4(1H)-one

The compound of Example 23 was obtained from the Korea Compound Bank.

(ChemBank#: 3595)

<Example 24> 3-acetyl-6-chloro-2-(3-chlorophenylamino)-8-nitroquinolin-4(1H)-one

The compound of Example 24 was obtained from the Korea Compound Bank.

(ChemBank#: 3542)

<Example 25> 3-acetyl-5,8-dichloro-2-(4-chloro-2-fluorophenylamino)quinolin-4(1H)-one

The compound of Example 25 was obtained from the Korea Compound Bank.

(ChemBank#: 3547)

<Example 26> 3-acetyl-5,8-dichloro-2-(4-chloro-2-(trifluoromethyl)phenylamino)quinolin-4(1H)-one

The compound of Example 26 was obtained from the Korea Compound Bank.

(ChemBank#: 3569)

<Example 27> 5,8-dichloro-2-(2,5-difluorophenylamino)-3-propionylquinolin-4(1H)-one

The compound of Example 27 was obtained from the Korea Compound Bank.

(ChemBank#: 3591)

<Example 28> 3-acetyl-6-chloro-2-(4-fluorophenylamino)-8-nitroquinolin-4(1H)-one

The compound of Example 28 was obtained from the Korea Compound Bank.

(ChemBank#: 3624)

<Example 29> 3-acetyl-6-chloro-2-(2-chloro-4-fluorophenylamino)-8-nitroquinolin-4(1H)-one

The compound of Example 29 was obtained from the Korea Compound Bank.

(ChemBank#: 3633)

<Example 30> 3-acetyl-8-chloro-2-(2,3-difluorophenylamino)-5-nitroquinolin-4(1H)-one

The compound of Example 30 was obtained from the Korea Compound Bank.

(ChemBank#: 4195)

<Example 31> 3-acetyl-2-(3,5-difluorophenylamino)-7,8-difluoroquinolin-4(1H)-one

The compound of Example 31 was obtained from the Korea Compound Bank.

(ChemBank#: 4171)

<Example 32> 3-acetyl-2-(4-bromophenylamino)-8-chloro-5-fluoroquinolin-4(1H)-one

The compound of Example 32 was obtained from the Korea Compound Bank.

(ChemBank#: 3828)

<Example 33> 3-acetyl-2-(4-chloro-2-fluorophenylamino)-5,8-difluoroquinolin-4(1H)-one

The compound of Example 33 was obtained from the Korea Compound Bank.

(ChemBank#: 229588)

<Example 34> 3-acetyl-2-(3,5-difluorophenylamino)-7,8-difluoroquinolin-4(1H)-one

The compound of Example 34 was obtained from the Korea Compound Bank.

(ChemBank#: 230226)

<Example 35> 3-acetyl-2-(2-chloro-4-methylphenylamino)-5,8-difluoroquinolin-4(1H)-one

The compound of Example 35 was obtained from the Korea Compound Bank.

(ChemBank#: 229587)

<Example 36> 3-acetyl-7,8-dichloro-2-(3,5-difluorophenylamino)quinolin-4(1H)-one

The compound of Example 36 was obtained from the Korea Compound Bank.

(ChemBank#: 229692)

<Example 37> 3-acetyl-8-chloro-5-fluoro-2-(2-fluoro-5-methylphenylamino)quinolin-4(1H)-one

The compound of Example 37 was obtained from the Korea Compound Bank.

(ChemBank#: 229661)

<Example 38> 5,8-dichloro-2-(2,4-difluorophenylamino)-3-propionylquinolin-4(1H)-one

The compound of Example 38 was obtained from the Korea Compound Bank.

(ChemBank#: 230113)

<Example 39> 3-acetyl-2-(4-bromophenylamino)-6,8-difluoroquinolin-4(1H)-one

The compound of Example 39 was obtained from the Korea Compound Bank.

(ChemBank#: 230184)

<Example 40> 3-acetyl-6-chloro-2-(2,4-dichlorophenylamino)-8-nitroquinolin-4(1H)-one

The compound of Example 40 was obtained from the Korea Compound Bank.

(ChemBank#: 3630)

<Example 41> 5,8-dichloro-2-(3,5-difluorophenylamino)-3-isobutyrylquinolin-4(1H)-one

The compound of Example 41 was obtained from the Korea Compound Bank.

(ChemBank#: 3916)

<Example 42> 3-acetyl-2-(4-bromophenylamino)-7,8-dichloroquinolin-4(1H)-one

The compound of Example 42 was obtained from the Korea Compound Bank.

(ChemBank#: 2418)

<Example 43> 3-acetyl-5,8-dibromo-2-(3,5-difluorophenylamino)quinolin-4(1H)-one

The compound of Example 43 was obtained from the Korea Compound Bank.

(ChemBank#: 3852)

<Example 44> 3-acetyl-5,8-difluoro-2-(4-(trifluoromethyl)phenylamino)quinolin-4(1H)-one

The compound of Example 44 was obtained from the Korea Compound Bank.

(ChemBank#: 229992)

The structures of the compounds of Examples 1 to 44 are summarized and shown in Table 1 below.

Example rescue Example rescue One

23

2

24

3

25

4

26

5

27

6

28

7

29

8

30

9

31

10

32

11

33

12

34

13

35

14

36

15

37

16

38

17

39

18

40

19

41

20

42

21

43

22

44

<Experimental Example 1> Antiviral activity evaluation

<1-1> Cell line and virus preparation

Vero cells used in the present invention were purchased and used from the American Type Culture Collection (ATCC, CCL-81; Manassas, VA), and 10% heat inactivated fetal bovine serum and 1x antibiotic-antifungal agents (Antibiotic-Antimycotic, Gibco/Thermo Fisher Scientific, Waltham, MA) in Dulbecco's modified Eagle's medium (DMEM; Welgene, Gyeongsan, Korea) and cultured at 37°C under 5% carbon dioxide.

MERS-CoV Korean isolate (MERS-CoV/KOR/KNIH/002_05_2015, Genbank accession no. KT029139.1 [Kim et al., 2015 doi:10.1128/genomeA.00787-15]) was provided by the National Institutes of Health, Korea Centers for Disease Control and Prevention. Taken, Kim et al., 2016 doi:10.1093 / cid / ciw239 according to the method presented in Vero cells proliferated. All experiments using MERS-CoV were conducted at the Pasteur Institute in Korea, which complied with the National Institutes of Health's enhanced biosafety level 3 (BL-3) containment procedure approved by the Korea Centers for Disease Control and Prevention.

<1-2> Reagent preparation

Chloroquine diphosphate (CQ; C6628) and lopinavir (LPV; GP6351)) were purchased from SelleckChem (Houston, TX) and Glentham Life Science (UK), respectively. The anti-MERS-CoV spike antibody used as the primary antibody was manufactured by Sino Biological Inc. (Beijing, China). The secondary antibody Alexa Fluor 488 goat anti-rabbit IgG and the nuclear chromosome Hoechst 33342 were purchased from Molecular Probes/ Thermo Fisher Scientific (Waltham, MA). 32% Paraformaldehyde (PFA) aqueous solution and normal chlorine serum were obtained from Electron Microscopy Sciences (Hatfield, PA) and Vector Laboratories, Inc. (Burlingame, CA).

<1-3> Image-based assay using immunofluorescence assay

Since cells infected with MERS-CoV express viral proteins, they can be measured using antibodies that specifically bind to viral proteins. In the present invention, cells were stained using an antibody that binds to the spike protein of MERS-CoV, and the infected cells were imaged through a microscope. The infection rate (number of cells expressing the MERS-CoV spike protein/total number of cells) was measured with an internally developed Image Mining 3.0 (IM 3.0) plug-in. In order to compare the antiviral effects of the small molecule compounds, infected cells treated with dimethylsulfoxide (DMSO) were used as a negative control, and two compounds (CQ and LPV) with known antiviral activity against MERS-CoV were used as positive controls. Was used to optimize the image-based assay.

<1-4> small molecular compound library

About 200,000 small molecular compounds were dissolved in DMSO and stored at -80°C until analysis.

<1-5> Image-based small molecular compound screening

Vero cells at 1.2 x 10 ⁴ cells per well in Opti-PRO™ SFM containing 4 mM L-Glutamine and 1 x Antibiotic-Antimycotic in black, 384-well, microclear plates (Clear plates, Greiner bio-one, Kremsmunster, Austria). After 24 hours, the low molecular weight compound was added to each well using an automated liquid handling system (Apricot Designs, Covina, CA) before viral infection. The final concentration of the test compound was 2.5 to 28.2 μM, and the concentration of DMSO was maintained at 0.5%. The compound-treated group was transferred to the BL-3 containment chamber and then infected with 0.0625 MOI of MERS-CoV.

PFA was used 24 hours after infection (final PAF concentration = 4%) to fix the infection. Anti-MERS-CoV spike antibody was treated on the immobilized cells and stained with Alexa Fluor 488 goat anti-rabbit IgG and Hoechst 33342. After fixation and staining of the infected cells, they were imaged on a fluorescence imaging system (Perkin Elmer Operetta, 20x, Waltham, MA) at 20x magnification. The infection rate for MERS-CoV of the cells treated with the small molecule compound was converted into a negative control group (0% infection inhibition rate) and a positive control group (100% infection inhibition rate) on each plate, and the small molecule causing an inhibitory effect of 90% or more. The compound was identified.

<1-6> Concentration-response curve experiment of effective compound

* The inhibitory effect of viral infection depending on the concentration of the compound can be found through the concentration-response curve experiment. The maximum concentration of the test compound is 5 mM, diluted 2 times using DMSO, and serially diluted up to step 10. This is treated on the prepared cells in the same way as in <1-5>. The maximum concentration of the final concentration of the experimental compound was 25 μM, and the concentration of DMSO was maintained at 0.5%. The compound treated group was transferred to the BL-3 containment chamber and then infected with 0.0625 MOI of MERS-CoV. After 24 hours of infection, the infection rate was imaged and converted in the same manner as in <1-5>. In the concentration response curve experiment, 50% virus inhibitory concentration (IC ₅₀ ) and 50% cytotoxicity concentration (CC ₅₀ ) of the compound were calculated.

The IC ₅₀ value, which is the concentration of the compound that inhibits the infection rate of the virus to 50%, is shown in Table 2 below.

Example IC ₅₀ (μM) Example IC ₅₀ (μM) One 1.23 23 4.18 2 0.015 24 1.64 3 0.266 25 1.14 4 0.066 26 1.52 5 0.157 27 0.496 6 0.354 28 1.83 7 0.156 29 1.98 8 2.93 30 0.847 9 1.06 31 0.136 10 0.243 32 0.842 11 0.591 33 0.627 12 0.219 34 0.588 13 0.189 35 2.83 14 0.788 36 0.781 15 0.556 37 4.39 16 0.443 38 1.1 17 0.484 39 1.13 18 0.231 40 0.756 19 0.774 41 0.119 20 0.489 42 0.455 21 1.44 43 0.311 22 6.36 44 2.62

As shown in Table 2 above,

Since the compound according to an aspect of the present invention has excellent antiviral inhibitory activity against MERS-CoV, it can be seen that it can be used as a treatment for MERS.

<Formulation Example 1> Preparation of pharmaceutical formulation

1-1. Preparation of powder

500 mg of the compound of formula 1

100 mg lactose

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

1-2. Manufacture of tablets

500 mg of the compound of formula 1

Corn starch 100 mg

100 mg lactose

2 mg of magnesium stearate

After mixing the above ingredients, tablets are prepared by tableting according to a conventional tablet preparation method.

1-3. Preparation of capsules

500 mg of the compound of formula 1

Corn starch 100 mg

100 mg lactose

2 mg of magnesium stearate

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare a capsule.

1-4. Preparation of injections

500 mg of the compound of formula 1

Suitable amount of sterile distilled water for injection

proper amount of pH adjuster

It is prepared with the above ingredients per ampoule (2 ml) according to a conventional injection preparation method.

1-5. Preparation of liquid

100 mg of the compound of formula 1

10 g of isomerized sugar

5 g of mannitol

Purified water appropriate amount

According to the usual preparation method of liquid preparation, add and dissolve each component in purified water, add lemon scent, mix the above ingredients, add purified water, add purified water, adjust the total to 100 ml, and fill in a brown bottle. The liquid is prepared by sterilization.

Claims (12)

A compound represented by the following Formula 1, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof:
[Formula 1]

(In Formula 1,
R ¹ is independently -H, -OH, halo, -NO ₂ , -NH ₂ , -CN, C _1-10 straight or branched chain alkyl unsubstituted or substituted with one or more halogens, or unsubstituted or one or more Halogen-substituted C _1-10 linear or branched alkoxy;

R ² is unsubstituted or C _1-10 straight or branched chain alkyl substituted with one or more halogens, or C _3-10 unsubstituted or substituted C _3-10 cycloalkyl;

R ³ is independently -H, -OH, halo, -NO ₂ , -NH ₂ , -CN, straight or branched C _1-10 alkyl unsubstituted or substituted with one or more halogens, or unsubstituted or one or more Halogen-substituted C _1-10 linear or branched alkoxy;

n is an integer from 0 to 4; And
m is an integer from 0 to 5).
The method of claim 1,
R ¹ is independently -H, -OH, halo, -NO ₂ , -NH ₂ , -CN, C _1-5 straight or branched chain alkyl unsubstituted or substituted with one or more halogens, or unsubstituted or one or more Halogen-substituted C _1-5 straight or branched alkoxy;

R ² is methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, tert-butyl, or cyclopropyl;

R ³ is independently -H, -OH, halo, -NO ₂ , -NH ₂ , -CN, straight or branched C _1-5 alkyl unsubstituted or substituted with one or more halogens, or unsubstituted or one or more Halogen-substituted C _1-5 straight or branched alkoxy;

n is an integer from 0 to 4; And
m is an integer of 0 to 4, a compound, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.
The method of claim 1,
R ¹ is independently -H, -F, -Cl, -Br, -NO ₂ , -CF ₃ , methyl or ethyl;
R ² is methyl, ethyl, or iso-propyl;
R ³ is independently -H, -F, -Cl, -Br, -CF ₃ , methyl, or methoxy;
n is an integer from 0 to 3; And
m is an integer of 0 to 3, a compound, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.
The method of claim 1,
The compound represented by Formula 1 is a compound, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof, characterized in that it is any one compound selected from the following compound group:
(1) 3-acetyl-8-chloro-5-methyl-2-(2,3,4-trifluorophenylamino)quinolin-4(1H)-one;
(2) 3-acetyl-6,8-difluoro-2-(2,3,4-trifluorophenylamino)quinolin-4(1H)-one;
(3) 3-acetyl-6-chloro-2-(3,5-dichlorophenylamino)-8-nitroquinolin-4(1H)-one;
(4) 3-acetyl-5,6,8-trichloro-2-(3-fluorophenylamino)quinolin-4(1H)-one;
(5) 6,8-difluoro-3-isobutyryl-2-(2,3,4-trifluorophenylamino)quinolin-4(1H)-one;
(6) 5,8-dichloro-2-(2,4-dichlorophenylamino)-3-isobutyrylquinolin-4(1H)-one;
(7) 5,8-dichloro-3-isobutyryl-2-(2,3,4-trifluorophenylamino)quinolin-4(1H)-one;
(8) 5,8-dichloro-3-isobutyryl-2-(4-methoxyphenylamino)quinolin-4(1H)-one;
(9) 3-acetyl-2-(2,4-difluorophenylamino)-6,8-difluoroquinolin-4(1H)-one;
(10) 3-acetyl-2-(2,4-dichlorophenylamino)-6,8-difluoroquinolin-4(1H)-one;
(11) 3-acetyl-6-chloro-2-(2,4-dichlorophenylamino)-8-nitroquinolin-4(1H)-one;
(12) 3-acetyl-5,8-dichloro-2-(2,3,4-trifluorophenylamino)quinolin-4(1H)-one;
(13) 5,8-dichloro-2-(3,5-difluorophenylamino)-3-isobutyrylquinolin-4(1H)-one;
(14) 2-(2,4-difluorophenylamino)-6,8-difluoro-3-isobutyrylquinolin-4(1H)-one;
(15) 5,8-dichloro-2-(4-chloro-2-fluorophenylamino)-3-isobutyrylquinolin-4(1H)-one;
(16) 3-acetyl-5,8-dichloro-2-(2,4-dichlorophenylamino)quinolin-4(1H)-one;
(17) 3-acetyl-5,8-dichloro-2-(2,4-difluorophenylamino)quinolin-4(1H)-one;
(18) 3-acetyl-8-chloro-5-(trifluoromethyl)-2-(2,3,4-trifluorophenylamino)quinolin-4(1H)-one;
(19) 3-acetyl-8-chloro-2-(2,4-dichlorophenylamino)-5-(trifluoromethyl)quinolin-4(1H)-one;
(20) 3-acetyl-2-(2,4-dichlorophenylamino)-5,8-difluoroquinolin-4(1H)-one;
(21) 3-acetyl-2-(4-chlorophenylamino)-6,8-difluoroquinolin-4(1H)-one;
(22) 3-acetyl-2-(2,4-dichlorophenylamino)-8-ethylquinolin-4(1H)-one;
(23) 3-acetyl-2-(2,5-difluorophenylamino)-5,8-difluoroquinolin-4(1H)-one;
(24) 3-acetyl-6-chloro-2-(3-chlorophenylamino)-8-nitroquinolin-4(1H)-one;
(25) 3-acetyl-5,8-dichloro-2-(4-chloro-2-fluorophenylamino)quinolin-4(1H)-one;
(26) 3-acetyl-5,8-dichloro-2-(4-chloro-2-(trifluoromethyl)phenylamino)quinolin-4(1H)-one;
(27) 5,8-dichloro-2-(2,5-difluorophenylamino)-3-propionylquinolin-4(1H)-one;
(28) 3-acetyl-6-chloro-2-(4-fluorophenylamino)-8-nitroquinolin-4(1H)-one;
(29) 3-acetyl-6-chloro-2-(2-chloro-4-fluorophenylamino)-8-nitroquinolin-4(1H)-one;
(30) 3-acetyl-8-chloro-2-(2,3-difluorophenylamino)-5-nitroquinolin-4(1H)-one;
(31) 3-acetyl-2-(3,5-difluorophenylamino)-7,8-difluoroquinolin-4(1H)-one;
(32) 3-acetyl-2-(4-bromophenylamino)-8-chloro-5-fluoroquinolin-4(1H)-one;
(33) 3-acetyl-2-(4-chloro-2-fluorophenylamino)-5,8-difluoroquinolin-4(1H)-one;
(34) 3-acetyl-2-(3,5-difluorophenylamino)-7,8-difluoroquinolin-4(1H)-one;
(35) 3-acetyl-2-(2-chloro-4-methylphenylamino)-5,8-difluoroquinolin-4(1H)-one;
(36) 3-acetyl-7,8-dichloro-2-(3,5-difluorophenylamino)quinolin-4(1H)-one;
(37) 3-acetyl-8-chloro-5-fluoro-2-(2-fluoro-5-methylphenylamino)quinolin-4(1H)-one;
(38) 5,8-dichloro-2-(2,4-difluorophenylamino)-3-propionylquinolin-4(1H)-one;
(39) 3-acetyl-2-(4-bromophenylamino)-6,8-difluoroquinolin-4(1H)-one;
(40) 3-acetyl-6-chloro-2-(2,4-dichlorophenylamino)-8-nitroquinolin-4(1H)-one;
(41) 5,8-dichloro-2-(3,5-difluorophenylamino)-3-isobutyrylquinolin-4(1H)-one;
(42) 3-acetyl-2-(4-bromophenylamino)-7,8-dichloroquinolin-4(1H)-one;
(43) 3-acetyl-5,8-dibromo-2-(3,5-difluorophenylamino)quinolin-4(1H)-one; And
(44) 3-acetyl-5,8-difluoro-2-(4-(trifluoromethyl)phenylamino)quinolin-4(1H)-one.
As shown in Scheme 1 below,
Preparing a compound represented by Formula 3 from the compound represented by Formula 2;
Preparing a compound represented by Formula 4 from the compound represented by Formula 3 prepared in the above step;
Preparing a compound represented by Formula 5 from the compound represented by Formula 4 prepared in the above step;
Preparing a compound represented by Formula 6 from the compound represented by Formula 5 prepared in the above step; And
Preparing a compound represented by Formula 1 from the compound represented by Formula 6 prepared in the above step; A method for preparing a compound represented by Formula 1 of claim 1, including:
[Scheme 1]

(In the above Scheme 1,
R ¹ , R ² , R ³ , n and m are independently as defined in Formula 1 of claim 1).
An antiviral pharmaceutical composition containing the compound represented by Formula 1 of claim 1, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
The method of claim 6,
The antiviral pharmaceutical composition includes corona virus, influenza virus, Alpui virus, ring virus, bovine diarrhea virus, chikungunya virus, dengue virus, hepatitis B virus, hepatitis C virus, human cytomegalovirus, human immunodeficiency Virus, Ilheus virus, Japanese encephalitis virus, Cocovera virus, Kunjin virus, Kiasanur forest disease virus, Leap disease virus, Measles virus, Metapneumovirus, Mosaic virus, Murray Valley virus, Parainfluenza virus, Polio virus, Powasan Antiviral pharmaceuticals having antiviral activity against at least one selected from the group consisting of viruses, respiratory syncytial virus, Roshio virus, St. Louis encephalitis virus, tick-mediated encephalitis virus, West Nile virus, and yellow fever virus. Ever composition.
The method of claim 7,
The coronaviruses include infectious bronchitis virus, bovine coronavirus, canine coronavirus, feline coronavirus, porcine erythropoietin encephalomyelitis virus, mouse hepatitis virus, turkey coronavirus, infectious gastrointestinal disease virus, rat coronavirus, human coronavirus 229E (HCoV-229E). ), human coronavirus NL63 (HCoV-NL63), human coronavirus OC43 (HCoV-OC43), SARS coronavirus (SARS-CoV), and Middle East Respiratory Syndrome coronavirus (MERS-Cov). Antiviral pharmaceutical composition, characterized in that.
The method of claim 6,
The antiviral pharmaceutical composition is used for the prevention or treatment of diseases caused by Middle East Respiratory Syndrome Coronavirus (MERS-Cov) infection.
A pharmaceutical composition for the prevention or treatment of MERS containing the compound represented by Formula 1 of claim 1, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
A health functional food composition for preventing or improving diseases caused by viral infection, containing the compound represented by Formula 1 of claim 1, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
The method of claim 11,
Health functional food composition, characterized in that the disease caused by the virus infection is MERS.