KR102188708B1 - 2-phenylchroman-4-one derivatives and antiviral composition comprising the same - Google Patents

Abstract

The present invention relates to a 2-phenylchroman-4-one derivative or a pharmaceutically acceptable salt thereof, a preparation method thereof, and a treatment for MERS containing the same as an active ingredient.

Description

2-phenylchroman-4-one derivatives and antiviral composition comprising the same}

The present invention relates to a 2-phenylchroman-4-one derivative or a pharmaceutically acceptable salt thereof, a preparation method thereof, and an antiviral therapeutic agent containing the same as an active ingredient.

The MERS virus belongs to the beta coronavirus and is a positive single-stranded RNA virus with 11 ORFs. Genetically, it has a very close relationship with the SARS coronavirus, a severe acute respiratory syndrome (SARS) pathogen that caused a total of 8,098 confirmed cases and 774 deaths worldwide in 2003. MERS virus is known to bind to human DPP4 (Dipeptidyl peptidase 4) receptor using S protein to infiltrate the cell [Wang N, Shi X, Jiang LJ, Zhang S, Wang D, Tong P, Guo D, et al. Structure of MERS-CoV spike receptor-binding domain complexed with human receptor DPP4. Cell Research. 2013:23:986]. On the basis of the outbreak of MERS outbreaks in the Middle East and Korea, the path of infection between animals (camels) and humans, infection routes between humans, and nosocomial infections have been identified [ www.cdc.gov/sars/about/fs-SARS. html ; HY Park, EJ Lee, YW Ryu, et al. Epidemiological investigation of MERS-CoV spread in a single hospital in South Korea, May to June 2015, EuroSurveill. 2015;20 (25): pii=21169]. There are currently no MERS vaccines and treatments approved domestically and internationally.

It is generally recommended to use drugs such as interferon, an immunomodulatory factor, and antiviral drugs, such as Ribarvirin or lopinavir. However, in the case of interferon and ribavirin, there are reports of concern about safety as they cause side effects such as decreased bone marrow function, anemia, and viral mutations. In addition, in the monkey model, co-administration of interferon and ribavirin showed the effect of treating MERS, but in actual clinical trials, it did not show a great effect on MERS patients, so the development of a safer and more effective MERS treatment is required.

The 2-phenylchroman-4-one derivative is the basic skeleton of bavachin and bavachnin, which are extracted from the seeds of a plant called Bogolji (or Pagoji, Psoralea corylifolia L.), which is used as a medicinal herb. It is known that barbakin and barbakinin exhibit various pharmacological activities, especially anticancer activity [S. Kuntz, U. Wenzel, H. Daniel, Eur. J. Nutr. 38 (1999) 133-142; P. Song, X.Z. Yang, J.Q. Yuan, J. Asian Nat. Prod. Res. 15 (2013) 624-630], anti-inflammatory activity [S.W. Lee, B.R. Yun, M.H. Kim, C.S. Park, W.S. Lee, H.M. Oh, M.C. Rho, Planta Med. 78 (2012) 903-906], Alzheimer's inhibitory activity [X. Chen, Y. Yang, Y. Zhang, FEBS Lett. 587 (2013) 2930-2935], and immunomodulatory effects [M.L. Sharma, B. Singh, B.K. Chandan, A. Khajuria, A. Kaul, S. Bani, S.K. Banerjee, S.S. Gambhir, Phytomedicine 3 (1996) 191-195] have been reported.

Synthetically, a method for synthesizing barbakinin in a racemic state has been reported [G. Du, L. Feng, Z. Yang, J. Shi, C. Huang, F. Guo, B. Li, W. Zhu, Bioorganic & Medicinal Chem. Lett. 25 (2015), 2579-2583; G. Du, Y. Zhuo, L. Feng, Z. Yang, J. Shi, C. Huang, B. Li, F. Guo, W. Zhu, Y. Li, ChemMedChem 2017, 183-193]. In addition, a patent [US 10,034,853 B2 (2018, 07, 31)] evaluating the efficacy of a PPAR agonist by synthesizing various derivatives using barbakin and barbakinin as starting materials and a paper evaluating anticancer effects [N. Gupta, A. Qayum, R. Raina, S. Gailora, S. Singh, P.L. Sangwan, European Journal of Medicinal Chemistry 145 (2018) 511-523] and the like have been reported. Synthesized and known 2-phenylchroman-4-one derivatives are in the case where X is oxygen in the representative structure, and there are no reports of derivatives in which X is nitrogen.

The present inventors have found that the 5-(1,3-diaryl-1H-pyrazol-4-ylmethylene)-thiazolidine-2,4-dione derivative has a selective inhibitory activity against CDC25B, and other similar dephosphorylation enzymes The present invention was completed by finding that it is a novel compound having selectivity and anticancer effect.

In addition, the inventors of the present invention that as 2-phenylchroman-4-one derivatives, natural products such as barbakin and barbakinin and their derivatives, and newly synthesized 2-phenylchroman-4-one derivatives are compounds having an inhibitory effect against MERS Revealed to complete the present invention.

An object of the present invention is to provide a 2-phenylchroman-4-one derivative or a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide a method for preparing a 2-phenylchroman-4-one derivative or a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide a composition for treating MERS comprising a 2-phenylchroman-4-one derivative or a pharmaceutically acceptable salt thereof.

The present invention provides a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

<Formula 1>

In addition, the present invention provides a pharmaceutical composition for preventing and treating coronavirus comprising the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

As another example, the present invention provides a pharmaceutical composition for the prevention and treatment of Middle East Respiratory Syndrome (MERS) coronavirus.

In addition, the pharmaceutical composition of the present invention may further include one or more pharmaceutically acceptable carriers, and may further include one or more known antiviral agents.

As another example, the present invention provides a health food composition for preventing and improving coronavirus infection, comprising the compound according to Formula 1 or a food pharmaceutically acceptable salt thereof as an active ingredient.

The compound containing the 2-phenylchroman-4-one derivative of Formula 1 and a pharmaceutically acceptable salt thereof according to the present invention has excellent proliferation inhibitory effect against MERS CoV infected with Vero cell line and less toxicity to Vero cell line. It can be used as a treatment for MERS.

In the present specification, the term'alkyl' refers to an aliphatic hydrocarbon radical, and includes all linear or branched hydrocarbon radicals. For example, C1-C6 alkyl is an aliphatic hydrocarbon having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, n-butyl, n-pentyl, n-hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl , Neopentyl, isopentyl, etc. are all included.

In addition, the term'alkoxy' refers to a radical in which a hydrogen atom of a hydroxy group is substituted with alkyl, unless otherwise defined, for example, C1-C6 alkoxy is methoxy, ethoxy, propoxy, n-butoxy, n -Pentyloxy, isopropoxy, sec-butoxy, tert-butoxy, neopentyloxy, isopentyloxy and the like are all included.

In addition, the term'aryl' refers to one or more hydrocarbon rings having a shared pi electron system, and includes hydrocarbon rings having 6 to 12 carbon atoms such as phenyl, naphthyl, and biphenyl.

The present invention provides a compound of Formula 1 or a pharmaceutically acceptable salt thereof.

The compound of Formula 1 or a pharmaceutically acceptable salt thereof may have a substituent containing an asymmetric atom, and in this case, the compound of Formula 1 or a salt thereof may include (R), (S), or racemic (RS). It can exist as an optical isomer. Accordingly, unless otherwise indicated, the compound of Formula 1 or a pharmaceutically acceptable salt thereof includes all optical isomers such as (R), (S), or racemic (RS).

The compound of Formula 1 of the present invention may be in the form of a pharmaceutically acceptable salt. The salts are conventional acid addition salts, for example, salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, or nitric acid, and acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, maleic acid, hydroxy Maleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanylic acid, 2-acetoxy-benzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid, oxalic acid, trifluoroacetic acid, citric acid, lactic acid, tartaric acid, coumaric acid, alginic acid , Camphorsulfonic acid, capric acid, myristic acid, hyporic acid, or salts derived from organic acids such as orotic acid. In addition, the salt is sodium hydroxide, lithium hydroxide, potassium hydroxide, calcium hydroxide, potassium carbonate, calcium carbonate, potassium t-butoxide, sodium ethoxide, triethylamine, ammonia, guanidine, ethylenediamine, ethanolamine, diethanol Salts derived from amines, triethanolamine, peperazine, morpholine, and dicyclohexylamine.

A pharmaceutically acceptable salt of the compound of Formula 1 can be prepared from the compound of Formula 1 by a conventional method. In general, the salt is prepared by reacting the compound of formula 1 in free acid/base form with a stoichiometric amount or an excess of the desired salt-forming inorganic acid, inorganic salt, organic acid or organic salt in a suitable solvent or various combinations of solvents. I can.

The present invention provides a pharmaceutical composition for preventing or treating coronavirus, comprising a therapeutically effective amount of the compound of Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

The pharmaceutical composition may contain a pharmaceutically acceptable carrier such as an excipient, a disintegrant, a sweetener, a lubricant or a flavoring agent, which are commonly used, and tablets, capsules, powders, granules, and suspensions according to a conventional method, Oral preparations such as emulsions or syrups; Or it may be formulated as a formulation for parenteral administration such as injection. The formulation can be formulated in a variety of forms, for example, a single dosage form or a multiple dosage form.

The composition of the present invention may be administered orally, or parenterally, including intravenous, intramuscular, intraperitoneal, subcutaneous, rectal and topical administration. The composition of the present invention can be preferably administered orally. Accordingly, the composition of the present invention may be formulated in various forms such as tablets, capsules, aqueous solutions or suspensions. In the case of tablets for oral use, carriers such as lactose and corn starch, and lubricants such as magnesium stearate may be usually added. In the case of capsules for oral administration, lactose and/or dried corn starch may be used as a diluent. If oral aqueous suspensions are required, the active ingredient can be combined with emulsifying and/or suspending agents. If necessary, certain sweetening and/or flavoring agents can be added. For intramuscular, intraperitoneal, subcutaneous and intravenous administration, sterile solutions of the active ingredient are usually prepared, and the pH of the solution must be appropriately adjusted and buffered. For intravenous administration, the total concentration of the solute should be adjusted to impart isotonicity to the formulation. The composition according to the present invention may be in the form of an aqueous solution comprising a pharmaceutically acceptable carrier such as saline with a pH of 7.4. The solution may be introduced into the patient's intramuscular bloodstream by local bolus injection.

The pharmaceutical composition according to the present invention can be administered in a therapeutically effective amount. Accordingly, the compound of Formula 1 or a pharmaceutically acceptable salt thereof contained in the pharmaceutical composition may be administered to a patient in an effective amount of about 10 mg/kg to about 500 mg/kg per day. Of course, the dose may be changed according to the patient's age, weight, sensitivity, symptoms, or drug efficacy of the compound.

The present invention also comprises administering a therapeutically effective amount of the compound of Formula 1 or a pharmaceutically acceptable salt thereof to mammals, including humans, of a virus, preferably a coronavirus, more preferably a MERS. Provides a method of prevention or treatment.

The present invention also provides the use of the compound of Formula 1 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for preventing or treating viruses.

Hereinafter, the present invention will be described in more detail through Examples and Test Examples. However, these Examples and Test Examples illustrate the present invention, and the present invention is not limited thereto.

In one aspect, the present invention provides a 2-phenylchroman-4-one derivative of the following formula (1) or a pharmaceutically acceptable salt thereof.

<Formula 1>

(In Formula 1,

X is NH or O,

R ₁ is hydrogen; C ₁ -C ₆ alkyl group; C ₂ -C ₆ alkenyl group; C ₃ -C ₆ alkynyl group; C ₁ -C ₆ haloalkyl group; C ₂ -C ₆ alkoxyalkyl group; C ₆ -C ₁₂ aryl group; And C ₁ -C ₆ carboxyalkyl group; is selected from the group consisting of,

R ₂ is hydrogen; C ₁ -C ₆ alkyl group; C ₂ -C ₆ alkenyl group; C ₃ -C ₆ alkynyl group; C ₁ -C ₆ haloalkyl group; C ₂ -C ₆ alkoxyalkyl group; And C ₆ -C ₁₂ aryl group; is selected from the group consisting of,

A-B and C-D are each independently a single bond (CH-CH) or a double bond (C=C) hydrocarbon.)

The compound of formula 1 of the present invention includes pharmaceutically acceptable salts thereof. Such salts can be prepared using conventional techniques in the art. Such pharmaceutically acceptable salts include, for example, hydrochloride, hydrobromide, sulfate, sulfamate, phosphate, nitrate, acetate, propionate, succinate, glycolate, stearate, maleate, hydroxy Maleate, phenylacetate, glutamate, benzoate, salicylate, sulfanylate, 2-acetoxy-benzoate, fumarate, toluenesulfonate, methanesulfonate, ethanesulfonate, oxalate, trifluoroacetic acid Salts, citrate, lactate, tartrate, coumaric acid salt, alginate, camposulfonic acid salt, capric acid salt, myristic acid salt, hyporic acid salt, or orotate salt, and other pharmaceutically acceptable salts. .

Among the compounds of the present invention, compounds in which X is oxygen can be obtained by a known method [G. Du, L. Feng, Z. Yang, J. Shi, C. Huang, F. Guo, B. Li, W. Zhu, Bioorganic & Medicinal Chem. Lett. 25 (2015), 2579-2583] compound of formula 2 synthesized using [barbakinin, 2-(4-hydroxyphenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl)chroman -4-one] as a basic material, a method of substituting a hydroxy group, a method of oxidizing the 2,3-position, and a method of reducing the side chain at the 6-position were used to synthesize various derivatives.

Compounds in which X is nitrogen among the compounds of the present invention are novel compounds, the compound of Formula 3 synthesized using a new synthesis method [2-(4-aminophenyl)-7-methoxy-6-(3-methylbut-2-en-) Using 1-yl)chroman-4-one] as a basic substance, various derivatives were synthesized using a method of substituting a hydroxy group, a method of oxidizing the 2,3-position, and a method of reducing the side chain at the 6-position. .

<Formula 2>

<Formula 3>

Schematic of the manufacturing method of the present Formula 3 can be represented by the following Scheme 1.

<Reaction Scheme 1>

First, N-BOC-4-aminobenzaldehyde is prepared by replacing 4-aminobenzaldehyde with a protecting group BOC group. Notification method [G. Du, L. Feng, Z. Yang, J. Shi, C. Huang, F. Guo, B. Li, W. Zhu, Bioorganic & Medicinal Chem. Lett. 25 (2015), 2579-2583] synthesized from 1-(2-hydroxy-4-methoxy-5-(3-methylbut-2-en-1-yl)phenyl)ethan-1-one and synthesized N- By reacting BOC-4-aminobenzaldehyde in the presence of an appropriate base, 3-(4-N-BOC-amino)phenyl)-1-(2-hydroxy-4-methoxy-5-(3-methylbut-2-en-1-) When yl)phenyl)prop-2-en-1-one is obtained and then heated in the presence of a weak base (eg potassium fluoride), the compound of formula 3 is obtained. Solvents, bases, and protecting groups used in these reactions can be synthesized using chemically similar solvents, bases, and protecting groups in addition to the conditions indicated in the text.

As another aspect of the present invention, the present invention relates to a treatment for MERS comprising a 2-phenylchroman-4-one derivative of Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

It was confirmed that the 2-phenylchroman-4-one derivative of Formula 1 effectively inhibited the MERS virus in a drug efficacy test using Vero cells infected with the MERS virus, and had low cytotoxicity, so that the treatment effect of MERS could be expected.

Therefore, the MERS treatment or pharmaceutical composition containing the compound of Formula 1 as an active ingredient according to the present invention is a conventional non-toxic pharmaceutically acceptable carrier, adjuvant or excipient, etc., added to the compound of Formula 1 Formulations, such as tablets, capsules, troches, solutions, and suspensions can be formulated for oral administration.

In addition, the dosage of the compound of Formula 1 to the human body may vary depending on the patient's age, weight, sex, dosage form, health condition, and disease level, and is generally 10 based on an adult patient weighing 70 kg. To 4,000 mg/day, and may be administered in divided doses from once a day to several times a day at regular time intervals according to the judgment of a doctor or pharmacist.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are only intended to aid understanding of the present invention, and the scope of the present invention is not limited to the following examples.

Example 1: Preparation of 7-methoxy-2-(4-(methoxymethoxy)phenyl)-6-(3-methylbut-2-en-1-yl)chromen-4-one (Compound No. 1)

(i) Preparation of 4-(methoxymethoxy)benzaldehyde

After dissolving 4-hydroxybenzaldehyde (200 mmol) in 500 mL DMF, DIPEA (220 mmol) and MOMCl (220 mmol) were added to the solution. The reaction mixture was stirred at room temperature for one day. The mixture was extracted with EtOAc and dried over anhydrous magnesium sulfate. Then, the mixture was filtered, and the organic solution was evaporated under reduced pressure to obtain 4-(methoxymethoxy)benzaldehyde as a yellow solid.

(Yield: 84%), yellow solid

¹ H NMR (300 MHz, Chloroform- d ) δ 9.92 (s, 1H), 7.91-7.77 (m, 2H), 7.22-7.11 (m, 2H), 5.27 (s, 2H), 3.51 (s, 3H) .

(ii) Preparation of 1-(4-methoxy-2-((3-methylbut-2-en-1-yl)oxy)phenyl)ethan-1-one

After dissolving 2-hydroxy-4-methoxyacetophenone (90 mmol) in acetone, potassium carbonate and prenyl bromide (135 mmol) were added to the solution. The reaction mixture was refluxed and stirred for one day. The mixture was extracted with EtOAc and dried over anhydrous magnesium sulfate. After that, the mixture was filtered, and the organic solution was evaporated under reduced pressure to form a yellow solid 1-(4-methoxy-2-((3-methylbut-2-en-1-yl)oxy)phenyl)ethan-1-one. Got it.

( Yield: 75%), yellow solid

¹ H NMR (300 MHz, Chloroform- d ) δ 7.82 (d, J = 8.7 Hz, 1H), 6.53-6.41 (m, 2H), 5.49 (ddt, J = 8.0, 6.6, 1.4 Hz, 1H), 4.58 (d, J = 6.7 Hz, 2H), 3.82 (s, 3H), 2.57 (s, 3H), 1.83-1.72 (m, 7H).

(iii) Preparation of 1-(2-hydroxy-4-methoxy-5-(3-methylbut-2-en-1-yl)phenyl)ethan-1-one

After dissolving 1-(4-methoxy-2-((3-methylbut-2-en-1-yl)oxy)phenyl)ethan-1-one in diethylaniline, the reaction mixture was refluxed and stirred for 4 hours. The mixture was extracted with EtOAc and dried over anhydrous magnesium sulfate. After that, the mixture was filtered and the organic solution was evaporated under reduced pressure. Purify the residue by column chromatography on silica to obtain 1-(2-hydroxy-4-methoxy-5-(3-methylbut-2-en-1-yl)phenyl)ethan-1-one as a yellow oil. Got it,

(Yield: 80%), yellow oil

(iv) (E)-1-(2-hydroxy-4-methoxy-5-(3-methylbut-2-en-1-yl)phenyl)-3-(4-(methoxymethoxy) phenyl)prop-2- Preparation of en-1-one

1-(2-hydroxy-4-methoxy-5-(3-methylbut-2-en-1-yl)phenyl)ethan-1-one (167 mmol) and 4-(methoxymethoxy)benzaldehyde (167 mmol) in 400 After dissolving in mL of ethanol, potassium trimethylsilanolate (554 mmol) was added to the solution. The reaction mixture was refluxed and stirred for 3 hours. After the reaction was terminated by adding a saturated solution of NH ₄ Cl to the mixture, it was extracted with ether and dried over anhydrous magnesium sulfate. Then, the mixture was filtered, and the organic solution was evaporated under reduced pressure. Purify the residue by column chromatography on silica (n-hexane / ether) to obtain a yellow solid (E)-1-(2-hydroxy-4-methoxy-5-(3-methylbut-2-en-1-yl) )phenyl)-3-(4-(methoxymethoxy) phenyl)prop-2-en-1-one was obtained.

(Yield: 50%), yellow solid

(v) 7-methoxy-2-(4-(methoxymethoxy)phenyl)-6-(3-methylbut-2-en-1-yl)chroman-4-one (Compound 1)

(E)-1-(2-hydroxy-4-methoxy-5-(3-methylbut-2-en-1-yl)phenyl)-3-(4-(methoxymethoxy) phenyl)prop-2-en-1 After -one (5.0 mmol) was dissolved in 50 mL of methanol, potassium fluoride (17 mmol) was added to the solution, and the reaction mixture was refluxed and stirred for 8 hours. The mixture was extracted with EtOAc and dried over anhydrous magnesium sulfate. After that, the mixture was filtered and the organic solution was evaporated under reduced pressure. Purify the residue by column chromatography on silica (n-hexane / EtOAc) to obtain 7-methoxy-2-(4-(methoxymethoxy)phenyl)-6-(3-methylbut-2-en-1-yl as a yellow oil. ) obtained chroman-4-one.

(Yield: 60%), yellow oil

Example 2: Preparation of 2-(4-hydroxyphenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl)chroman-4-one (Compound No. 2)

After dissolving 7-methoxy-2-(4-(methoxymethoxy)phenyl)-6-(3-methylbut-2-en-1-yl)chroman-4-one (Compound 1) in methanol, 3 N HCl was added to the solution. Added to. The reaction mixture was refluxed and stirred for 30 minutes. After the reaction was terminated by adding distilled water to the mixture, the mixture was extracted with EtOAc and dried over anhydrous magnesium sulfate. After that, the mixture was filtered and the organic solution was evaporated under reduced pressure. Purification of the residue by column chromatography on silica (n-hexane / EtOAc) as a yellow solid 2-(4-hydroxyphenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl)chroman- 4-one (Compound No. 2) was obtained.

(Yield: 90%), yellow solid

NMR (300 MHz, Chloroform-d) δ 7.68 (s, 1H), 7.35 (d, J = 8.4 Hz, 2H), 6.89 (d, J = 8.4 Hz, 2H), 6.44 (s, 1H), 5.38 ( dd, J = 13.2, 2.7 Hz, 1H), 5.27 (t, d = 7.5 Hz, 1H), 5.20 (s, 1H), 3.84 (s, 3H), 3.24 (d, J = 7.2 Hz, 2H), 3.08-2.98 (m, 1H), 2.77 (dd, J = 17.1, 3.0 Hz, 1H), 1.71 (d, J = 13.5 Hz, 3H)

Example 3: Preparation of 7-methoxy-2-(4-methoxyphenyl)-6-(3-methylbut-2-en-1-yl)chroman-4-one (Compound No. 3)

2-(4-hydroxyphenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl)chroman-4-one (Compound No. 2) (0.12 mmol) and potassium carbonate (0.24 mmol) was dissolved in 0.3 mL of DMF, and iodomethane (0.17 mmol) was added to the solution. The reaction mixture was stirred at room temperature for one day. The mixture was extracted with EtOAc and dried over anhydrous magnesium sulfate. After that, the mixture was filtered and the organic solution was evaporated under reduced pressure. Purify the residue by column chromatography on silica (n-hexane / EtOAc) to obtain 7-methoxy-2-(4-methoxyphenyl)-6-(3-methylbut-2-en-1-yl)chroman- as a transparent oil. 4-one (Compound No. 3) was obtained.

(Yield: 68%), transparent oil

NMR (300 MHz, Chloroform-d) δ 7.68 (s, 1H), 7.46-7.33 (m, 7H), 7.03 (d, J = 9.0 Hz, 2H), 6.44 (s, 1H), 5.39 (dd. J = 12, 3.0 Hz, 1H), 5.30-5.24 (m, 1H), 5.10 (s, 2H), 3.84 (s, 3H), 3.24 (d, J = 6.0 Hz, 2H), 3.09-2.99 (m, 1H), 2.77 (dd, J = 18, 3.0 Hz, 1H), 1.72 (d, J = 12 Hz, 6H)

Example 4: Preparation of 2-(4-(benzyloxy)phenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl)chroman-4-one (Compound No. 4)

2-(4-hydroxyphenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl)chroman-4-one (Compound No. 2) (0.12 mmol) and potassium carbonate (0.24 mmol) was dissolved in 0.3 mL of DMF, and benzyl bromide (0.17 mmol) was added to the solution. The reaction mixture was stirred at 60° C. for one day. The mixture was extracted with EtOAc and dried over anhydrous magnesium sulfate. After that, the mixture was filtered and the organic solution was evaporated under reduced pressure. The residue was purified by column chromatography on silica (n-hexane / EtOAc) to obtain 2-(4-(benzyloxy)phenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl as a white solid. )chroman-4-one (Compound No. 4) was obtained.

(Yield: 49%), white solid

NMR (300 MHz, Chloroform-d) δ7.68 (s, 1H), 7.46-7.33 (m, 7H), 7.03 (d, J = 8.7 Hz, 2H), 6.44 (s, 1H), 5.39 (dd, J = 13.2, 3 Hz, 1H), 5.30-5.24 (m, 1H), 5.10 (s, 2H), 3.84 (s, 3H), 3.24 (d, J = 7.2 Hz, 2H), 3.09-2.99 (m , 1H), 2.77 (dd, J = 17.1, 3 Hz, 1H), 1.72 (d, J = 13.5, 6H)

Example 5: Preparation of 2-(4-isopropoxyphenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl)chroman-4-one (Compound No. 5)

2-(4-hydroxyphenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl)chroman-4-one (Compound No. 2) (0.12 mmol) and potassium carbonate (0.24 mmol) was dissolved in 0.3 mL of DMF, and 2-bromopropane (0.17 mmol) was added to the solution. The reaction mixture was stirred at 60° C. for one day. The mixture was extracted with EtOAc and dried over anhydrous magnesium sulfate. After that, the mixture was filtered and the organic solution was evaporated under reduced pressure. Purify the residue by column chromatography on silica (n-hexane / EtOAc) to obtain a white solid 2-(4-isopropoxyphenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl)chroman- 4-one (Compound No. 5) was obtained.

(Yield: 31%), white solid

NMR (300 MHz, Chloroform-d) δ 7.68 (s, 1H), 7.38 (d, J = 9.0 Hz, 2H), 6.93 (d, J = 6 Hz, 2H), 6.45 (s, 1H), 3.38 ( dd, J = 15, 3.0 Hz, 1H), 5.30-5.25 (m, 1H), 4.62-4.53 (m, 1H), 3.84 (s, 3H), 3.24 (d, J = 6Hz, 2H), 3.10- 2.99 (m, 1H), 2.77 (dd, J = 12, 3.0 Hz, 1H), 1.72 (d, J = 12 Hz. 6H), 1.35 (d, J = 6 Hz, 6H)

Example 6: Preparation of 2-(4-hydroxyphenyl)-6-isopentyl-7-methoxychroman-4-one (Compound No. 6)

After dissolving 2-(4-hydroxyphenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl)chroman-4-one (Compound No. 2) (1.5 mmol) in 3 mL of ethanol, 10 wt% Pd/C was added to the solution. The reaction mixture was stirred at room temperature for one day under hydrogen gas. After the mixture was filtered through Celite, the organic solution was evaporated under reduced pressure. The residue was purified by column chromatography on silica (n-hexane / EtOAc) to obtain 2-(4-hydroxyphenyl)-6-isopentyl-7-methoxychroman-4-one as a white solid.

(Yield: 56%), white solid

¹ H NMR (300 MHz, Chloroform- d ) δ 7.05-7.08 (d, 2H), 6.82 (s, 1H), 6.74-6.77 (d, 2H), 6.34 (s, 1H) 4.63 (s, 1H) 4.47 (s, 1H) 3.76 (s, 3H) 2.47-2.63 (m, 4H), 1.83-1.93 (m, 2H), 1.36-1.44 (m, 2H) 0.91-0.94 (d, 6H).

Example 7: Preparation of 6-isopentyl-7-methoxy-2-(4-methoxyphenyl)chroman-4-one (Compound No. 7)

(Yield: 26%), transparent oil

After dissolving 2-(4-hydroxyphenyl)-6-isopentyl-7-methoxychroman-4-one (compound number 6) (0.059 mmol) and potassium carbonate (0.11 mmol) in 0.2 mL of DMF, iodomethane ( 0.088 mmol) was added to the solution. The reaction mixture was stirred at 60° C. for one day. The mixture was extracted with EtOAc and dried over anhydrous magnesium sulfate. After that, the mixture was filtered and the organic solution was evaporated under reduced pressure. The residue was purified by column chromatography on silica (n-hexane / EtOAc) to obtain 6-isopentyl-7-methoxy-2-(4-methoxyphenyl)chroman-4-one (Compound No. 7) as a transparent oil.

(Yield: 26%), transparent oil

NMR (300 MHz, Chloroform-d) δ7.12 (d, J = 8.7 Hz, 1H), 7.07 (d, J=8.4 Hz, 1H), 6.84 (d, J = 7.8 Hz, 2H), 6.74 (d , J = 8.4 Hz, 1H), 6.42 (s, 1H), 6.34 (s, 1H), 4.45 (s, 1H), 3.18 (d, J = 3.0 Hz, 3H), 3.78 (d, J = 10.2 Hz , 3H), 2.64-2.48 (m, 2H), 1.94-1.81 (m, 2H), 1.62-1.53 (m, 2H), 1.45-1.37 (m, 2H)

Example 8: Preparation of 2-(4-(benzyloxy)phenyl)-6-isopentyl-7-methoxychroman-4-one (Compound No. 8)

2-(4-hydroxyphenyl)-6-isopentyl-7-methoxychroman-4-one (compound number 6) (0.059 mmol) and potassium carbonate (0.11 mmol) were dissolved in 0.2 mL of DMF, and then benzyl bromide (0.088 mmol) was added to the solution. The reaction mixture was stirred at 60° C. for one day. The mixture was extracted with EtOAc and dried over anhydrous magnesium sulfate. After that, the mixture was filtered and the organic solution was evaporated under reduced pressure. The residue was purified by column chromatography on silica (n-hexane / EtOAc) to obtain 2-(4-(benzyloxy)phenyl)-6-isopentyl-7-methoxychroman-4-one (Compound No. 8) as a transparent oil. .

(Yield: 13%), transparent oil

NMR (300 MHz, Chloroform-d) δ7.45-7.32 (m, 5H), 7.11 (d, J = 8.4 Hz, 1H), 7.03 (d, J = 8.4 Hz, 1H), 6.90 (d, J = 12.8 Hz, 1H), 6.87 (m, 2H), 6.71 (d, J = 8.4 Hz, 1H), 6.58 (s, 1H), 5.04 (s, 2H), 4.50 (s, 1H), 3.77 (d, J = 3.6 Hz, 3H), 2.64-2.47 (m, 2H), 1.94-1.85 (m, 2H), 1.61-1.51 (m, 2H), 1.44-1.38 (m, 2H), 0.93 (d, J = 6.3 Hz, 6H)

Example 9: Preparation of compound number 9

After dissolving 2-(4-hydroxyphenyl)-6-isopentyl-7-methoxychroman-4-one (compound number 6) (0.059 mmol) and potassium carbonate (0.11 mmol) in 0.2 mL of DMF, 2- Bromopropane (0.088 mmol) was added to the solution. The reaction mixture was stirred at 60° C. for one day. The mixture was extracted with EtOAc and dried over anhydrous magnesium sulfate. After that, the mixture was filtered and the organic solution was evaporated under reduced pressure. The residue was purified by column chromatography on silica (n-hexane / EtOAc) to give compound No. 9 as a transparent oil.

(Yield: 25%), transparent oil

NMR (300 MHz, Chloroform-d) δ7.08 (t, J = 8.4 Hz, 2H), 6.83 (t, J = 5.7 Hz, 2H), 6.74 (d, J = 8.4 Hz, 1H), 6.34 (s , 1H), 4.57-4.43 (m, 2H), 3.77 (d, J = 7.8 Hz, 3H), 2.63-2.47 (m, 2H), 1.94-1.79 (m, 2H), 1.59-1.53 (m, 2H) ), 1.44-1.67 (m, 2H), 1.33 (d, J = 6.0 Hz, 6H), 0.93 (d, J = 6.6 Hz, 6H)

Example 10: Preparation of 2-(4-hydroxyphenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl)-4H-chromen-4-one (Compound No. 10)

2-(4-hydroxyphenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl)chroman-4-one (Compound No. 2) (0.15 mmol) in 1 mL of pyridine After dissolving, iodine (0.22 mmol) was added to the solution. The reaction mixture was stirred at 100° C. for 5 hours. The mixture was extracted with EtOAc and dried over anhydrous magnesium sulfate. After that, filter the mixture. The organic solution was evaporated under reduced pressure. Purification of the residue by column chromatography on silica (n-hexane / EtOAc) as a white solid 2-(4-hydroxyphenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl)-4H -chromen-4-one (compound number 10) was obtained.

(Yield: 37%), white solid

¹ H NMR (300 MHz, DMSO- d ) δ 10.25 (s, 1H), 7.92 (d, J = 8.4 Hz, 2H), 7.68 (s, 1H), 7.26 (s, 1H) 6.92 (d, J = 8.4 Hz, 2H), 6.75 (s, 1H), 5.28 (t, J = 7.5 Hz, 1H), 3.94 (s, 3H), 3.26 (d, J= 9 Hz, 2H), 1.68 (d, J = 15 Hz 6H).

Example 11: 7-methoxy-2-(4-methoxyphenyl)-6-(3-methylbut-2-en-1-yl)-4H-chromen-4-one

(Yield: 32%), white solid

Compound (14) (0.053 mmol) and potassium carbonate (0.11 mmol) were dissolved in 0.2 mL of DMF, and iodomethane (0.080 mmol) was added to the solution. The reaction mixture was stirred at room temperature for 3 hours. The mixture was extracted with EtOAc and dried over anhydrous magnesium sulfate. After that, the mixture was filtered and the organic solution was evaporated under reduced pressure. The residue was purified by column chromatography on silica (n-hexane / EtOAc) to obtain a white solid compound (15) .

Example 12: Preparation of 2-(4-aminophenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl)-4H-chromen-4-one (Compound 12)

(i) tert-butyl (4-formylphenyl)carbamate

4-aminobezaldehyde (25 mmol) was dissolved in 50 mL of DMF and then lowered to 0 °C, and potassium hydroxide (KOH) (75 mmol) and di-tert-butyl dicarbonate (50 mmol) were added to the solution. The reaction mixture was stirred at 60° C. for 4 hours. The mixture was extracted with EtOAc and dried over anhydrous magnesium sulfate. Thereafter, the mixture was filtered, and the organic solution was evaporated under reduced pressure to obtain tert-butyl (4-formylphenyl)carbamate as a yellow solid.

(Yield: 85%), yellow solid

(ii) (E)-3-(4-((tert-butoxymethyl)amino)phenyl)-1-(2-hydroxy-4-methoxy-5-(3-methylbut-2-en-1-yl)phenyl )prop-2-en-1-one

1-(4-methoxy-2-((3-methylbut-2-en-1-yl)oxy)phenyl)ethan-1-one (8.5 mmol) and tert-butyl (4-formylphenyl)carbamate ( 8.5 mmol) was dissolved in 25 mL of ethanol, and potassium trimethylsilanolate (30 mmol) was added to the solution. The reaction mixture was refluxed and stirred for one day. After the reaction was terminated by adding a saturated NH ₄ Cl solution to the mixture, extraction was performed using ether, and dried over anhydrous magnesium sulfate. Then, the mixture was filtered, and the organic solution was evaporated under reduced pressure. Purify the residue by column chromatography on silica (n-hexane / EtOAc) as a yellow solid (E)-3-(4-((tert-butoxymethyl)amino)phenyl)-1-(2-hydroxy-4- methoxy-5-(3-methylbut-2-en-1-yl)phenyl)prop-2-en-1-one was obtained.

(Yield: 47%), yellow solid

¹ H NMR (300 MHz, Chloroform- d ) δ 13.61 (s, 1H), 7.82 (d, J = 15 Hz, 1H), 7.59 (s, 1H), 7.49 (d, J = 8.4 Hz, 2H), 7.41 (s, 1H), 7.36 (s, 1H), 6.70 (d, J = 8.4 Hz, 2H), 6.43 (s, 1H), 5.28 (t, J = 7.2 Hz, 1H), 4.02 (brs, 2H ), 3.87 (s, 3H), 3.26 (d, J = 7.2 Hz, 2H), 1.77 (d, J = 11 Hz, 6H).

(iii) 2-(4-aminophenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl)chroman-4-one (Compound 12)

(E)-3-(4-((tert-butoxymethyl)amino)phenyl)-1-(2-hydroxy-4-methoxy-5-(3-methylbut-2-en-1-yl)phenyl)prop- 2-en-1-one (0.14 mmol) was dissolved in 1 mL of methanol, and potassium fluoride (0.49 mmol) was added to the solution. The reaction mixture was refluxed and stirred for one day. The mixture was extracted with EtOAc and dried over anhydrous magnesium sulfate. After that, the mixture was filtered and the organic solution was evaporated under reduced pressure. The residue was purified by column chromatography on silica (n-hexane / EtOAc) to form a yellow solid 2-(4-aminophenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl)-4H -chromen-4-one (compound 12) was obtained.

(Yield: 37%), yellow solid

¹ H NMR (300 MHz, Chloroform- d ) δ 7.67 (s, 1H), 7.26 (d, J = 8.4 Hz, 2H), 6.72 (d, J = 8.4 Hz, 2H), 6.44 (s, 1H), 5.35-5.25 (m, 1H), 3.83 (s, 3H), 3.24 (d, J = 7.2 Hz, 2H), 3.11-2.71 (m, 2H), 1.72 (d, J = 13 Hz, 6H).

Example 13: Preparation of N-(4-(7-methoxy-6-(3-methylbut-2-en-1-yl)-4-oxochroman-2-yl)phenyl)acetamide (Compound 13)

2-(4-aminophenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl)-4H-chromen-4-one (Compound 12) (0.059 mmol) dissolved in 0.5 mL of DCM Then, triethylamine (0.12 mmol) and acetic anhydride (0.065 mmol) were added to the solution. The reaction mixture was stirred at room temperature for 2 hours. The mixture was extracted using dichloromethane and dried over anhydrous magnesium sulfate. After that, the mixture was filtered and the organic solution was evaporated under reduced pressure. The residue was purified by column chromatography on silica (n-hexane / EtOAc) as a yellow solid N-(4-(7-methoxy-6-(3-methylbut-2-en-1-yl)-4-oxochroman) -2-yl)phenyl)acetamide (compound 13) was obtained,

(Yield: 79%), yellow solid

¹ H NMR (300 MHz, Chloroform- d ) δ 7.67 (s, 1H), 7.57 (d, J = 8.4 Hz, 2H), 7.43 (d, J = 8.4 Hz, 2H), 6.45 (s, 1H), 5.41 (dd, J = 13, 3.0 Hz, 1H), 5.27 (t, J = 9.9 Hz, 1H), 3.85 (s, 3H), 3.24 (d, J = 7.2 Hz, 2H), 3.05-2.75 (m , 2H), 2.20 (s, 3H), 1.71 (d, J = 14 Hz, 6H).

Example 14: 7-methoxy-6-(3-methylbut-2-en-1-yl)-2-(4-(prop-2-yn-1-ylamino)phenyl)chroman-4-one (Compound 14 ) Of manufacture

2-(4-aminophenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl)-4H-chromen-4-one (Compound 12) (0.05 mmol) dissolved in 0.5 mL of dichloromethane Then, triethylamine (0.12 mmol) and propargyl chloride (0.065 mmol) were added to the solution. The reaction mixture was stirred at room temperature for one day. The mixture was extracted using dichloromethane and dried over anhydrous magnesium sulfate. After that, the mixture was filtered and the organic solution was evaporated under reduced pressure. Purify the residue by column chromatography on silica (n-hexane / EtOAc) to obtain 7-methoxy-6-(3-methylbut-2-en-1-yl)-2-(4-(prop-2) as an orange solid. -yn-1-ylamino)phenyl)chroman-4-one (Compound 14) was obtained.

(Yield: 33%), orange solid

¹ H NMR (300 MHz, Chloroform- d ) δ 7.67 (s, 1H), 7.32 (d, J = 8.4 Hz, 2H), 6.73 (d, J = 8.4 Hz, 2H), 6.44 (s, 1H), 5.37-5.24 (m, 2H), 3.96 (d, J = 2.4 Hz, 2H), 3.83 (s, 1H), 3.24 (d, J = 7.5 Hz, 2H), 3.11-2.72 (m, 2H), 1.72 (d, J = 13 Hz, 6H).

Example 15: Preparation of 2-(4-aminophenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl)-4H-chromen-4-one (Compound 15)

2-(4-aminophenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl)chroman-4-one (Compound 12) (0.30 mmol) dissolved in 3 mL of pyridine Then iodine (0.44 mmol) was added to the solution. The reaction mixture was stirred at 100 °C for 2 hours. The mixture was extracted with EtOAc and dried over anhydrous magnesium sulfate. After that, filter the mixture. The organic solution was evaporated under reduced pressure. Purify the residue by column chromatography on silica (n-hexane / EtOAc) to obtain 2-(4-aminophenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl)-4H as an orange solid. -chromen-4-one (compound 15) was obtained.

(Yield: 37%), orange solid

¹ H NMR (300 MHz, Chloroform- d ) δ 7.94 (s, 1H), 7.73 (d, J = 8.4 Hz, 2H), 6.89 (s, 1H), 6.75 (d, J = 8.4 Hz, 2H), 6.63 (s, 1H), 5.33-5.30 (m, 1H), 3.95 (s, 3H), 3.36 (d, J = 7.5 Hz, 2H), 1.73 (d, J = 11 Hz, 6H).

Example 16: N-(4-(7-methoxy-6-(3-methylbut-2-en-1-yl)-4-oxo-4H-chromen-2-yl)phenyl)acetamide (Compound 16) Produce

2-(4-aminophenyl)-7-methoxy-6-(3-methylbut-2-en-1-yl)-4H-chromen-4-one (Compound 15) (0.060 mmol) dissolved in 0.5 mL of dichloromethane Then, triethylamine (0.12 mmol) and acetic anhydride (0.065 mmol) were added to the solution. The reaction mixture was stirred at room temperature for one day. The mixture was extracted using dichloromethane and dried over anhydrous magnesium sulfate. After that, the mixture was filtered and the organic solution was evaporated under reduced pressure. The residue was purified by column chromatography on silica (n-hexane / EtOAc) and N-(4-(7-methoxy-6-(3-methylbut-2-en-1-yl)-4-oxo) as a white solid. -4H-chromen-2-yl)phenyl)acetamide (Compound 16) was obtained.

(Yield: 60%), white solid

¹ H NMR (300 MHz, Chloroform- d ) δ 7.94 (s, 1H), 7.88 (d, J = 6.6 Hz, 2H), 7.69 (d, J = 8.7 Hz, 2H), 7.56 (s, 1H), 6.93 (s, 1H), 6.81 (s, 1H), 5.32 (t, J = 7.5 Hz, 1H), 3.97 (s, 3H), 3.37 (d, J = 7.5 Hz, 2H), 2.24 (s, 3H ), 1.73 (d, J = 13 Hz, 6H).

The compounds synthesized through the above examples have a racemic structure, unlike barbakin and barbakinin, which are obtained as natural products, and two optical isomers are mixed in a ratio of 1:1, and the chemical structure is listed in Table 1 below. As shown.

One

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

Example 17: MERS CoV inhibition experiment using Vero cell line

(i) cell lines and viruses

Vero cells used in the present invention were used by purchasing the American Type Culture Collection (ATCC, CCL-81; Manassas, VA), 10% heat inactivated fetal bovine serum and 1x Antibiotic-Antimycotic (Gibco/Thermo Fisher Scientific, Waltham, MA) in Dulbecco's modified Eagle's medium (DMEM; Welgene, Gyeongsan, Korea) and incubated 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]) from the Korea Centers for Disease Control and Prevention, National Institutes of Health It was provided and proliferated in vero cells according to the method suggested in prior literature (Kim et al., 2016 doi:10.1093/cid/ciw239). All experiments using MERS-CoV were carried out at the Pasteur Institute in Korea, which complied with the National Institute of Health's enhanced biosafety level 3 (BL-3) containment procedure approved by the Korea Centers for Disease Control and Prevention.

(ii) reagent

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 goat serum were obtained from Electron Microscopy Sciences (Hatfield, PA) and Vector Laboratories, Inc., respectively. (Burlingame, CA).

(iii) 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 effect of the small molecule compound, 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. The image-based assay was optimized.

(iv) storage of the compound

Compounds used in about 2 experiments were dissolved in DMSO and stored at -80 °C until analysis.

(v) Image-based small molecular compound screening

Vero cells were placed in each well of 1.2x10 ⁴ cells in Opti-PRO™ SFM containing 4 mM L-Glutamine and 1x Antibiotic-Antimycotic in black, 384-well, microclear plates (Clear plates, Greiner bio-one, Kremsmunster, Austria). After 24 hours, small molecule compounds were added to each well using an automated liquid handling system (Apricot Designs, Covina, CA) prior to viral infection. The final concentration of the test compound was ?μM, and the concentration of DMSO was maintained at 0.5%. The compound was transferred to the treated 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.

The MERS CoV inhibitory effect obtained through the above experiment and the cytotoxicity results for the Vero cell line are listed in Table 2 below.

Since the compounds of Formula 1 used for the drug efficacy experiment are racemics, two optical isomers are mixed in a ratio of 1:1, so if only one optical isomer has a drug effect, more than twice the drug efficacy can be expected.

Compound number MERS CoV inhibitory effect (μM) Vero cell line cytotoxicity (μM) One 10.0 >25 2 6.6 13.0 3 6.3 >25 4 10.0 >25 5 13.2 >25 6 3.3 4.0 7 11.1 15.2 8 6.7 15.0 9 5.6 12.8 10 4.8 >25 11 14.4 >25 12 11.6 >25 13 8.4 >25 14 12.0 >25 15 11.7 >25 16 8.7 >25

Claims (8)

A compound represented by Formula 1 or a pharmaceutically acceptable salt thereof.
<Formula 1>

(In Formula 1,
X is NH,
R ₁ is hydrogen; C ₁ -C ₆ alkyl group; C ₂ -C ₆ alkenyl group; And C ₃ -C ₆ alkynyl group; is selected from the group consisting of,
R ₂ is hydrogen; C ₁ -C ₆ alkyl group; C ₂ -C ₆ alkenyl group; And C ₃ -C ₆ alkynyl group; is selected from the group consisting of,
AB and CD are each independently a single bond (CH-CH) or a double bond (C=C) hydrocarbon.)
The method of claim 1, wherein the salt is hydrochloride, hydrobromide, sulfate, sulfamate, phosphate, nitrate, acetate, propionate, succinate, glycolate, stearate, maleate, hydroxymaleate, phenylacetate , Glutamate, benzoate, salicylate, sulfanylate, 2-acetoxy-benzoate, fumarate, toluenesulfonate, methanesulfonate, ethanesulfonate, oxalate, trifluoroacetate, citrate, lactic acid Salt, characterized in that it is selected from the group consisting of salts, tartaric acid salts, coumaric acid salts, alginates, camposulfonic acid salts, capric acid salts, myristic acid salts, hyporic acid salts and orotate acid salts
Any one of the compounds represented by the following formula

Middle East Respiratory Syndrome (MERS) containing a compound selected from the group consisting of compounds represented by the following formulae or a pharmaceutically acceptable salt thereof as an active ingredient for the prevention and treatment of coronavirus Composition

delete The pharmaceutical composition according to claim 4, further comprising at least one pharmaceutically acceptable carrier.
The pharmaceutical composition of claim 4, further comprising at least one antiviral agent.
Middle East Respiratory Syndrome (MERS) containing as an active ingredient a compound selected from the group consisting of compounds represented by the following formulas or compounds represented by the following formula, or for preventing and improving coronavirus infection Health food composition

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