KR102456293B1 - Cyclic sulfonamide derivatives and antiviral composition comprising the same - Google Patents

Abstract

The present invention relates to a cyclic sulfonamide derivative or a pharmaceutically acceptable salt thereof and an antiviral pharmaceutical composition containing the same as an active ingredient.

Description

Cyclic sulfonamide derivatives and antiviral composition comprising the same

The present invention relates to a cyclic sulfonamide derivative or a pharmaceutically acceptable salt thereof and an antiviral pharmaceutical composition containing the same as an active ingredient.

Coronavirus Infectious Disease-19 (COVID-19) is a respiratory syndrome caused by SARS-CoV-2 infection. The increase in the number of deaths is causing great social problems.

The SARS-CoV-2 pathogen is an RNA virus belonging to Coronaviridae. The SARS coronavirus (SARS-CoV), a pathogen of severe acute respiratory syndrome (SARS) in 2002, and the Middle East Respiratory Syndrome (MERS) in 2015 It is closely related to MERS-CoV, the pathogen of SARS-CoV-2 and SARS-CoV bind to ACE2 (Angiotensin Converting Enzyme2) receptor, and MERS-CoV utilizes DPP4 (Dipeptidyl Peptidase4; CD26) as a receptor. As a result of the 3D molecular structure analysis, it was confirmed that the spike proteins of SARS-CoV-2 and SARS-CoV have a fairly similar shape (Walls et al., 2020, Structure, Function and antigenicity of the SARS-CoV-2 spike). glycoprotein, Cell, DOI: 10.1016/j.cell.2020.02.058.; Wrapp et al., 2020, Cryo-EM structure of the 2019-nCoV spike in the prefusion conformation. Science.), due to this conformational similarity, SARS -CoV-2 was also found to strongly adhere to the surface of host cells through the ACE2 receptor, and the molecular structure of the spike protein and ACE2 conjugate was also recently revealed (Zhou et al., 2020 A pneumonia outbreak associated with a new coronavirus). of probable bat origin. Nature.; Yan et al., 2020, Structural basis for the recognition of the SARS-CoV-2 by full-length human ACE2. Science, DOI: 10.1126/science.abb2762). However, SARS-CoV-2 transmits and spreads faster than SARS-CoV, which allows SARS-CoV-2 to bind more strongly to ACE2 in the host cell and to allow a portion of the spike protein to be more easily cleaved by protein scissors. It is presumed that this is because it is deformed.

Research and development for prevention and treatment of COVID-19 is actively underway, and there is no treatment for COVID-19 that has been developed so far, so the development of a treatment is required.

The present inventors have completed the present invention by finding that cyclic sulfonamide derivatives introduced with various substituents are compounds having an inhibitory effect on coronavirus infection-19.

An object of the present invention is to provide a cyclic sulfonamide derivative or a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide an antiviral pharmaceutical composition comprising a cyclic sulfonamide 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 an antiviral pharmaceutical composition comprising the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

As another example, the present invention provides a pharmaceutical composition for coronavirus antiviral, wherein the coronavirus is SARS coronavirus (SARS-CoV), MERS coronavirus (MERS-CoV) or SARS coronavirus 2 (SARS-CoV2) to be.

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.

The composition containing a cyclic sulfonamide derivative or a pharmaceutically acceptable salt thereof according to the present invention has excellent proliferation inhibitory effect on SARS-CoV-2 infected with Vero cell line and has no toxicity to Vero cell line. It can be used as a therapeutic agent for viruses, and in particular, it can be used as a therapeutic agent for COVID-19 (coronavirus infection-19).

Hereinafter, the present invention will be described in detail. The terms or words used in the present specification and claims should not be construed as being limited to ordinary or dictionary meanings, and the inventor may properly define the concept of the term in order to best describe his invention. Based on the principle that there is, it should be interpreted as meaning and concept consistent with the technical idea of the present invention.

The terms used herein are briefly described.

The term “alkyl”, “alkyl group” or “alkylene group” refers to an aliphatic hydrocarbon radical and includes both straight-chain or branched hydrocarbon radicals. For example, C ₁ ~ C ₆ Alkyl is an aliphatic hydrocarbon having 1 to 6 carbon atoms, methyl, ethyl, propyl, n-butyl, n-pentyl, n-hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, neopentyl, isopentyl, and the like.

The term “alkene”, “alkenyl group” or “alkenylene group” refers to a group in which at least two carbon atoms consist of at least one carbon-carbon double bond, and “alkyne” or “alkynyl group” means that at least two carbon atoms are refers to a group consisting of at least one carbon-carbon triple bond.

The term “heterocycle” or “heterocyclic group” includes at least one of a single ring and multiple rings, and includes a heteroaliphatic ring and a heteroaromatic ring, unless otherwise specified. It may be formed by combining adjacent functional groups.

The term “aryl” or “aryl group” refers to one or more hydrocarbon rings having a shared pi electron system, and examples thereof include phenyl, naphthyl, biphenyl, and the like.

The term “halogen” is an element belonging to group 17 of the periodic table, and specifically, may be fluorine, chlorine, bromine, or iodine.

Unless otherwise defined, the term “alkoxy” or “alkoxy group” refers to a radical in which a hydrogen atom of a hydroxy group is substituted with alkyl, for example, C ₁ to C ₆ alkoxy is methoxy, ethoxy, propoxy, n- butoxy, n-pentyloxy, isopropoxy, sec-butoxy, tert-butoxy, neopentyloxy, isopentyloxy, and the like.

The term "hydroxy group" means -OH.

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

A compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof

[Formula 1]

In Formula 1, each symbol may be defined as follows.

1) X is O or -NOCH ₃ .

2) R ₁ is a C ₁ ~ C ₁₀ alkyl group; C ₂ ~ C ₁₀ Alkenyl group; C ₂ ~ C ₁₀ Alkynyl group; Or C ₆ ~ C ₂₄ Aryl group; and, most preferably C ₁ ~ C ₆ Alkyl group; C ₂ ~ C ₆ An alkenyl group; C ₂ ~ C ₆ Alkynyl group; Or C ₆ ~ C ₁₀ Aryl group; may be.

3) R ₂ is a hydroxyl group; Or a C ₁ ~ C ₁₀ alkoxy group; and, when R ₂ is an alkoxy group, most preferably a C ₁ ~ C ₆ alkoxy group.

4) L is a C ₁ ~ C ₃ alkylene group; Or C ₂ ~ C ₃ An alkenylene group;

5) R ₃ is a C ₆ ~ C ₂₄ aryl group; -NH-C ₆ ~ C ₂₄ Aryl group; Or O, N, S containing at least one hetero atom of C ₂ ~ C ₁₂ A heterocyclic group; and, most preferably, a C ₆ ~ C ₁₀ Aryl group; -NH-C ₆ ~ C ₁₀ Aryl group; Or O, N, S containing at least one hetero atom of C ₂ ~ C ₁₀ A heterocyclic group; may be.

6) each R ₄ is the same or different and is hydrogen; halogen; O, N, S containing at least one hetero atom C ₂ ~ C ₂₄ A heterocyclic group; Or -NR ^a R ^b ; and, when R ₄ is a heterocyclic group, most preferably a C ₂ ~ C ₁₂ heterocyclic group.

7) wherein R ^a and R ^b are each independently hydrogen; Or C ₁ ~ C ₁₀ Alkyl group;

8) a is an integer from 0 to 4.

9) Here, the alkyl group, the alkenyl group, the alkynyl group, the alkoxy group, the aryl group and the heterocyclic group are each deuterium; halogen; hydroxyl group; cyano group; nitro group; C ₁ ~ C ₆ Alkyl group; C ₂ ~ C ₆ An alkenyl group; C ₂ ~ C ₆ Alkynyl group; C ₁ ~ C ₆ Alkoxy group; C ₁ ~ C ₆ Alkyl group substituted with fluorine; C ₁ ~ C ₆ Alkoxy group substituted with fluorine; C ₆ ~ C ₁₂ Aryl group; And O, N, S C ₂ ~ C ₁₀ A heterocyclic group comprising at least one heteroatom; may be further substituted with one or more substituents selected from the group consisting of, these substituents are bonded to each other to form a ring Also, the 'ring' refers to a fused ring consisting of an aliphatic ring having 3 to 20 carbon atoms, an aromatic ring having 6 to 20 carbon atoms, a heterocycle having 2 to 20 carbon atoms, or a combination thereof, and includes a saturated or unsaturated ring.

In addition, the present invention includes a compound in which Formula 1 is represented by Formula 2 below.

[Formula 2]

In Formula 2, each symbol may be defined as follows.

1) X, R ₂ , R ₄ and a are the same as defined in Formula 1 above.

2) R ₅ and R ₆ are each the same or different, and each independently hydrogen; halogen; cyano group; C ₁ ~ C ₁₀ Alkyl group; C ₂ ~ C ₁₀ Alkenyl group; C ₂ ~ C ₁₀ Alkynyl group; And C ₁ ~ C ₁₀ Alkoxy group; is selected from the group consisting of.

When R ₅ and R ₆ are an alkyl group, most preferably a C ₁ ~ C ₆ alkyl group, most preferably a C ₂ ~ C ₆ alkenyl group in the case of an alkenyl group, and most preferably a C ₂ ~ C ₆ It is an alkynyl group, and in the case of an alkoxy group, it may be most preferably a C ₁ ~ C ₆ alkoxy group.

3) b and c are each independently an integer of 0 to 5;

In addition, the present invention includes a compound in which Chemical Formula 2 is represented by any one of Chemical Formulas 2-1 to 2-3 below.

[Formula 2-1] [Formula 2-2]

[Formula 2-3]

{In Formulas 2-1 to 2-3,

1) R ₄ and a are the same as defined in Formula 1 above,

2) R ₅ , R ₆ , b and c are the same as defined in Formula 2 above.}

In addition, the present invention includes compounds in which Chemical Formula 1 is represented by Chemical Formula 3 or Chemical Formula 4 below.

[Formula 3] [Formula 4]

{In Formulas 3 to 4,

1) R ₄ and a are the same as defined in Formula 1 above,

2) R ₆ and c are the same as defined in Formula 2 above.}

In addition, the present invention includes a compound in which Formula 1 is represented by Formula 5 below.

[Formula 5]

{In Formula 5,

1) R ₄ and a are the same as defined in Formula 1 above,

2) R ₅ and b are the same as defined in Formula 2 above,

3) R ₇ is hydrogen or halogen,

4) d is an integer from 0 to 5.}

In addition, the present invention includes a compound in which Formula 1 is represented by Formula 6 below.

[Formula 6]

{In Formula 6,

1) R ₄ and a are the same as defined in Formula 1 above,

2) R ₅ and b are the same as defined in Formula 2 above,

3) R ₈ is the same as the definition of R ₅ in Formula 2 above.}

Specifically, the compound represented by Formula 1 may be any one of the following compounds, but is not limited thereto.

The compound of Formula 1 or a pharmaceutically acceptable salt thereof may have a substituent including an asymmetric atom. In this case, the compound of Formula 1 or a salt thereof is (R), (S), or a racemic form (RS). It may exist as optical isomers, such as 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 include 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 acid Maleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic 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 , salts derived from organic acids such as camphorsulfonic acid, capric acid, myristic acid, hyporic acid or 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, or dicyclohexylamine.

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

The present invention provides an antiviral pharmaceutical composition 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 include pharmaceutically acceptable carriers such as excipients, disintegrants, sweeteners, lubricants or flavoring agents commonly used, and tablets, capsules, powders, granules and suspensions according to conventional methods; It may be formulated as an oral preparation such as an emulsion or syrup, or a preparation for parenteral administration such as an injection. The formulation may be formulated in a variety of forms, for example, single-dose or multiple-dose dosage forms.

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 may preferably be 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 oral tablets, 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 dry corn starch may be used as diluents. If an aqueous suspension for oral use is required, the active ingredient may be combined with emulsifying and/or suspending agents. If desired, certain sweetening and/or flavoring agents may be added. For intramuscular, intraperitoneal, subcutaneous and intravenous administration, a sterile solution of the active ingredient is usually prepared, and the pH of the solution must be appropriately adjusted and buffered. For intravenous administration, the total concentration of solutes should be adjusted to render the formulation isotonic. The composition according to the present invention may be in the form of an aqueous solution containing a pharmaceutically acceptable carrier, such as saline having a pH of 7.4. The solution can be introduced into the patient's intramuscular bloodstream by local bolus injection.

The pharmaceutical composition according to the present invention may 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, susceptibility, symptoms, or drug efficacy of the compound.

The present invention also provides a method of administering a therapeutically effective amount of a compound of Formula 1 or a pharmaceutically acceptable salt thereof to a mammal, including a human. It provides a method for preventing or treating Respiratory Syndrome (SARS), Middle East Respiratory Syndrome (MERS) or Coronavirus Infectious Disease-19.

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 antiviral agents.

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 the prevention or treatment of viruses.

As another aspect of the present invention, the present invention relates to a coronavirus, in particular, severe acute respiratory syndrome (SARS), Middle East comprising a cyclic sulfonamide derivative of Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. It relates to a treatment for respiratory syndrome (MERS) or coronavirus infection-19.

The cyclic sulfonamide derivative of Formula 1 effectively inhibits SARS-CoV-2 in a drug efficacy test using Vero cells infected with SARS-CoV-2 and has low cytotoxicity, so it can be expected to have a therapeutic effect on coronavirus. was confirmed, and in particular, it was confirmed that it can be used as a treatment for COVID-19.

Therefore, the coronavirus infection-19 therapeutic agent or pharmaceutical composition comprising the compound of Formula 1 according to the present invention as an active ingredient is a pharmaceutical field by adding a conventional non-toxic pharmaceutically acceptable carrier, adjuvant or excipient, etc. to the compound of Formula 1 In conventional formulations, for example, tablets, capsules, troches, solutions, suspensions, etc. can be formulated into formulations 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 status 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 once or several times a day at regular time intervals according to the judgment of a doctor or pharmacist.

Hereinafter, synthetic examples and examples of the compound represented by the formula according to the present invention will be described in detail, but the present invention is not limited to the following examples.

C1-11 Synthesis example

1) c-11-1 synthesis

Saccharin (1.20 g, 6.55 mmol) was dissolved in DMF (60 mL) and the substanceOne(1.81 g, 7.20 mmol) and triethylamine (1 mL, 7.20 mmol) were added, followed by stirring at room temperature for 9 hours. When the reaction is complete, the reaction mixture is diluted with ice water, the resulting solid is filtered, washed well with distilled water and hexane, and vacuum dried to c-11-1 (2.18 g, 94%) was obtained.

2) c-11-2 synthesis

c-11-1 (2.18 g, 6.16 mmol) was dissolved in absolute ethanol (40 mL), and 21% sodium ethoxide ethanol solution (6.9 mL, 18.49 mmol) was added dropwise. After the reaction mixture was stirred at 60° C. for 30 minutes, when the reaction was completed, 1N HCl was treated (pH=2~3). After dilution with ice water, the resulting solid was filtered, washed well with distilled water and hexane, and dried under vacuum to c-11-2 (1.59 g, 73%) was obtained.

3) Substance 2 Synthesis

Substance 2-1 (5 mL, 39.82 mmol) was dissolved in dichloromethane (250 mL), triethylamine (8.32 mL, 59.72 mmol) was added, and then chloroacetyl chloride (3.48 mL, 43.80 mmol) was slowly added dropwise at 0°C. did. After the reaction mixture is stirred at room temperature for 1 hour, when the reaction is completed, dichloromethane is added to dilute the solution, and the solution is washed with 2N HCl and water. After the organic layers were collected, magnesium sulfate was added, filtered, and the filtrate was concentrated to obtain a substance 2 (7.62 g, 80%) by silica gel chromatography (eluent ethyl acetate:hexane = 1:4).

4) C1-11 synthesis

c-11-2 (440 mg, 1.31 mmol) was dissolved in DMF (10 mL) and material 2 (374 mg, 1.57 mmol) and 60% NaH (63 mg, 1.57 mmol) were added. After the reaction mixture was stirred at room temperature for 3 hours, when the reaction was completed, saturated aqueous ammonium chloride solution (50 mL) was added at 0° C., and the mixture was extracted with ethyl acetate (40 mL × 3). Magnesium sulfate was added to the extracted organic layer, filtered, and the filtrate was concentrated, followed by silica gel chromatography (eluent ethyl acetate: hexane = 1:4) to C1-11. (268 mg, 39%) was obtained. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 14.59 (s, 1H), 10.30 (s, 1H), 8.24 - 8.15 (m, 1H), 8.15 - 8.00 (m, 2H), 8.00 - 7.87 (m , 3H), 7.69 (t, J = 9.1 Hz, 1H), 7.57 (d, J = 8.3 Hz, 2H), 7.45 (d, J = 8.4 Hz, 2H), 4.07 (s, 2H).

C1-30 Synthesis example

1) Substance 3-2 Synthesis

30% aqueous ammonium hydroxide solution (13 mL) was added to material 3-1 (2.8 g, 13.42 mmol) at 0° C. and the reaction mixture was heated to reflux at 100° C. for 1 hour. After cooling to room temperature, the resulting solid was filtered, washed well with distilled water and hexane, and dried under vacuum to obtain material 3-2 (2.5 g, 98%).

2) Substance 3-3 Synthesis

Material 3-2 (2.45 g, 11.74 mmol) was dissolved in 5% aqueous NaOH solution (70 mL) and KMnO ₄ (4.64 g, 29.36 mmol) was added slowly. The reaction mixture was heated and refluxed at 100°C for 5 hours. When the reaction was completed, it was treated with 1N HCl (pH = 1) and extracted with ethyl acetate (70 mL × 3). The organic layer was washed with distilled water and saturated aqueous NaCl solution, filtered with magnesium sulfate, and the filtrate was concentrated to obtain material 3-3 (2.16 g, 76%).

3) Synthesis of substance 3

Material 3-3 (2.25 g, 10.265 mmol) was dissolved in sulfuric acid (7.5 mL) and stirred for 1.5 hours. When the reaction was completed, ice and distilled water were added to filter the precipitated solid, and then washed with water to obtain material 3 (1.98 g, 96%).

4) Synthesis of C1-30

C1-26 (100 mg, 0.18 mmol) was dissolved in dimethyl sulfoxide (1.5 mL), and then K ₂ CO ₃ (73 mg, 0.53 mmol) and methyl piperazine (97 μL, 0.88 mmol) were added. The reaction mixture was stirred at 80° C. for 18 hours. After cooling to room temperature, C1-30 (47 mg, 41%) was obtained by high performance liquid chromatography. ¹ H NMR (500 MHz, DMSO- d ₆ ) δ 10.28 (s, 1H), 9.96 (s, 1H), 8.06 (dd, J = 16.1, 8.0 Hz, 3H), 7.65 (t, J = 8.9 Hz, 1H), 7.59 (d, J = 8.5 Hz, 2H), 7.48 (d, J = 8.5 Hz, 2H), 7.42 (dd, J = 9.1, 2.5 Hz, 1H), 7.37 (d, J = 2.6 Hz, 1H), 4.32 (d, J = 13.0 Hz, 2H), 4.01 (s, 2H), 3.56 (d, J = 12.3 Hz, 2H), 3.29 (s, 2H), 3.16 (s, 2H), 2.88 ( s, 3H).

C1-47 Synthesis example

1) c-47-2 synthesis

c-47-1 (1000 mg, 2.83 mmol) was dissolved in anhydrous dichloromethane (30 mL), and DABCO (32 mg, 0.28 mmol) and ethyl propiolate (315 μL, 3.11 mmol) were added at room temperature. The reaction mixture was heated to reflux for 3 hours. After cooling to room temperature, distillation under reduced pressure and silica gel chromatography (eluent ethyl acetate:hexane = 2:3) gave c-47-2 (887 mg, 69%).

2) c-47-3 synthesis

c-47-2 (500 mg, 1.11 mmol) was dissolved in THF (10 mL), and LiOH·H ₂ O (232 mg, 5.54 mmol) was dissolved in distilled water (10 mL) and added. The reaction mixture was stirred at 60° C. for 9 hours. The reaction mixture was diluted with distilled water (20 mL), treated with 2N HCl (pH=3), and extracted with ethyl acetate (20 mL × 3). The organic layer was filtered by adding magnesium sulfate, and the filtrate was concentrated to obtain c-47-3 (339 mg, 72%).

3) Synthesis of C1-47

c-47-3 (100 mg, 0.24 mmol) was dissolved in dichloromethane (2 mL), followed by 3-trifluoromethylaniline (38 μL, 0.28 mmol), EDCI (204 mg, 1.06 mmol) and DMAP (3 mg, 0.024 mmol) was added. The reaction mixture was stirred for 9 hours, diluted with dichloromethane, and washed with distilled water. The organic layer was filtered by adding magnesium sulfate, and the filtrate was concentrated to obtain C1-47 (13 mg, 9%) by high performance liquid chromatography. ¹ H NMR (500 MHz, DMSO- d ₆ ) δ 10.17 (s, 1H), 8.24 - 8.17 (m, 1H), 8.09 - 7.95 (m, 4H), 7.90 (s, 1H), 7.67 (dd, J = 17.8, 8.8 Hz, 2H), 7.46 (d, J = 13.5 Hz, 1H), 7.43 - 7.33 (m, 2H), 7.00 (d, J = 7.8 Hz, 1H), 5.56 (d, J = 13.5 Hz) , 1H).

C1-48 Synthesis example

C1-19 (200 mg, 0.36 mmol) was dissolved in pyridine (4 mL) and NH ₂ OCH ₃ (75 mg, 0.91 mmol) was added. The reaction mixture was stirred at 120° C. for 9 hours, the solvent was distilled under reduced pressure, and concentrated, and then C1-48 (36 mg, 17%) was obtained by high performance liquid chromatography.

C1-48 Synthesis example

C1-19 (50 mg, 0.09 mmol) was dissolved in DMF (1.5 mL) and K ₂ CO ₃ (25 mg, 0.18 mmol) and MeI (11 mL, 0.18 mmol) were added. The reaction mixture was stirred at 100° C. for 5 hours, cooled to room temperature, diluted with ethyl acetate, and washed with distilled water. The organic layer was filtered by adding magnesium sulfate, and the filtrate was concentrated to obtain C1-48 (21 mg, 41%) by high performance liquid chromatography. ¹ H NMR (500 MHz, Chloroform- d ) δ 8.77 (s, 1H), 7.98 - 7.92 (m, 3H), 7.84 - 7.70 (m, 3H), 7.52 - 7.45 (m, 3H), 7.38 - 7.28 ( m, 2H), 7.00 - 6.94 (m, 1H), 4.09 (s, 2H), 3.56 (s, 3H).

Table 1 below shows ¹ H-NMR values of compounds C1-1 to C1-52 according to the present invention.

compound ¹ H-NMR C1-1 ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 9.85 (s, 1H), 8.21 - 8.15 (m, 1H), 8.04 (d, J = 7.6 Hz, 2H), 7.94 - 7.83 (m, 3H), 7.70 (t, J = 7.3 Hz, 1H), 7.63 (t, J = 7.6 Hz, 2H), 7.18 (h, J = 7.2, 6.5 Hz, 4H), 6.96 (tt, J = 6.9, 1.6 Hz, 1H) ), 3.99 (s, 2H). C1-2 ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 10.08 (s, 1H), 8.24 - 8.11 (m, 1H), 8.03 (d, J = 7.5 Hz, 2H), 7.95 - 7.83 (m, 3H), 7.69 (t, J = 7.4 Hz, 1H), 7.62 (t, J = 7.6 Hz, 2H), 7.22 (td, J = 8.2, 6.7 Hz, 1H), 7.12 (d, J = 11.5 Hz, 1H), 6.96 (d, J = 8.2 Hz, 1H), 6.80 (td, J = 8.5, 2.6 Hz, 1H), 3.99 (s, 2H). C1-3 ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 15.11 (s, 1H), 10.16 (s, 1H), 8.23 - 8.13 (m, 1H), 8.08 - 7.98 (m, 2H), 7.91 (qd, J) = 6.2, 5.6, 3.4 Hz, 3H), 7.64 (ddd, J = 14.5, 7.9, 6.2 Hz, 3H), 7.37 - 7.24 (m, 2H), 7.22 - 7.14 (m, 1H), 7.01 - 6.91 (m) , 1H), 3.99 (s, 2H). C1-4 ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 15.08 (s, 1H), 10.07 (s, 1H), 8.23 - 8.14 (m, 1H), 8.03 (d, J = 7.4 Hz, 2H), 7.96 - 7.85 (m, 3H), 7.73 - 7.57 (m, 3H), 7.34 (d, J = 2.2 Hz, 1H), 7.22 (t, J = 8.0 Hz, 1H), 7.13 - 7.00 (m, 2H), 3.99 (s, 2H). C1-5 ¹ H NMR (300 MHz, Chloroform-d) δ 15.57 (s, 1H), 8.27 - 8.21 (m, 1H), 8.17 - 8.09 (m, 2H), 7.91 (dd, J = 7.5, 1.5 Hz, 1H) , 7.83 - 7.70 (m, 2H), 7.27 - 7.03 (m, 7H), 3.83 (s, 2H). C1-6 ¹ H NMR (300 MHz, Chloroform-d) δ 15.55 (s, 1H), 8.27 - 8.22 (m, 1H), 8.16 - 8.08 (m, 2H), 7.91 (dd, J = 7.5, 1.5 Hz, 1H) , 7.85 - 7.71 (m, 2H), 7.36 (s, 1H), 7.23 - 7.10 (m, 3H), 7.06 (dt, J = 10.7, 2.3 Hz, 1H), 6.84 - 6.71 (m, 2H), 3.83 (s, 2H). C1-7 ¹ H NMR (400 MHz, Chloroform-d) δ 15.52 (s, 1H), 8.22 (d, J = 7.7 Hz, 1H), 8.09 (dd, J = 8.5, 5.4 Hz, 2H), 7.88 (d, J) = 7.5 Hz, 1H), 7.83 - 7.65 (m, 2H), 7.37 (s, 1H), 7.23 (t, J = 8.2 Hz, 1H), 7.17 - 7.07 (m, 3H), 6.95 (dd, J = 17.9, 8.2 Hz, 2H), 3.80 (s, 2H). C1-8 ¹ H NMR (300 MHz, Chloroform-d) δ 15.51 (s, 1H), 8.26 - 8.19 (m, 1H), 8.14 - 8.05 (m, 2H), 7.89 (dd, J = 7.6, 1.5 Hz, 1H) , 7.83 - 7.69 (m, 2H), 7.48 (d, J = 8.7 Hz, 3H), 7.24 (d, J = 8.4 Hz, 2H), 7.16 - 7.07 (m, 2H), 3.83 (s, 2H). C1-9 ¹ H NMR (500 MHz, Chloroform-d) δ 8.23 (d, J = 7.7 Hz, 1H), 8.09 (dd, J = 6.9, 1.9 Hz, 1H), 8.06 - 8.00 (m, 1H), 7.89 (d) , J = 7.6 Hz, 1H), 7.80 (t, J = 7.6 Hz, 1H), 7.74 (t, J = 7.5 Hz, 1H), 7.31 (s, 1H), 7.17 (dt, J = 21.6, 8.2 Hz) , 3H), 7.05 (d, J = 7.9 Hz, 1H), 6.96 (d, J = 8.1 Hz, 1H), 3.81 (s, 2H). C1-10 ¹ H NMR (300 MHz, Chloroform-d) δ 15.43 (s, 1H), 8.28 - 8.22 (m, 1H), 8.11 (dd, J = 7.0, 2.2 Hz, 1H), 8.05 (ddd, J = 8.5, 4.6, 2.2 Hz, 1H), 7.91 (dd, J = 7.5, 1.5 Hz, 1H), 7.84 - 7.71 (m, 2H), 7.42 (s, 1H), 7.27 - 7.14 (m, 3H), 7.05 - 6.99 (m, 1H), 6.96 (d, J = 8.3 Hz, 1H), 3.84 (s, 2H). C1-11 ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 14.59 (s, 1H), 10.30 (s, 1H), 8.24 - 8.15 (m, 1H), 8.15 - 8.00 (m, 2H), 8.00 - 7.87 (m , 3H), 7.69 (t, J = 9.1 Hz, 1H), 7.57 (d, J = 8.3 Hz, 2H), 7.45 (d, J = 8.4 Hz, 2H), 4.07 (s, 2H). C1-12 ¹ H NMR (300 MHz, Chloroform-d) δ 15.48 (s, 1H), 8.29 - 8.21 (m, 1H), 7.91 (tt, J = 5.2, 1.2 Hz, 2H), 7.87 - 7.70 (m, 3H) , 7.48 (td, J = 8.0, 5.5 Hz, 1H), 7.32 (s, 1H), 7.23 (tdd, J = 8.3, 2.7, 1.0 Hz, 1H), 7.19 - 7.12 (m, 2H), 7.10 - 6.96 (m, 2H), 3.82 (s, 2H). C1-13 ¹ H NMR (400 MHz, Chloroform-d) δ 15.46 (s, 1H), 8.23 (d, J = 7.6 Hz, 1H), 7.88 (t, J = 7.5 Hz, 2H), 7.84 - 7.68 (m, 3H) ), 7.49 - 7.34 (m, 2H), 7.23 (t, J = 8.2 Hz, 1H), 7.18 (ddd, J = 10.1, 8.0, 2.6 Hz, 1H), 7.10 (s, 1H), 7.02 (d, J = 8.1 Hz, 1H), 6.93 (d, J = 8.3 Hz, 1H), 3.80 (s, 2H). C1-14 ¹ H NMR (300 MHz, Chloroform-d) δ 15.48 (s, 1H), 8.30 - 8.22 (m, 1H), 7.95 - 7.71 (m, 5H), 7.53 - 7.41 (m, 4H), 7.30 (s, 1H), 7.27 (s, 1H), 7.24 - 7.15 (m, 1H), 3.85 (s, 2H). C1-15 ¹ H NMR (300 MHz, Chloroform-d) δ 15.43 (s, 1H), 8.26 - 8.19 (m, 1H), 8.01 - 7.86 (m, 3H), 7.84 - 7.71 (m, 2H), 7.49 - 7.36 ( m, 2H), 7.29 (s, 1H), 7.17 - 7.10 (m, 2H), 7.07 - 6.95 (m, 2H), 3.79 (s, 2H). C1-16 ¹ H NMR (300 MHz, Chloroform-d) δ 15.46 (s, 1H), 8.28 - 8.22 (m, 1H), 8.03 - 7.96 (m, 2H), 7.92 (dd, J = 7.5, 1.5 Hz, 1H) , 7.85 - 7.71 (m, 2H), 7.50 - 7.35 (m, 3H), 7.24 (d, J = 8.2 Hz, 1H), 7.13 (s, 1H), 7.05 (ddd, J = 8.2, 2.1, 1.0 Hz) , 1H), 6.96 (ddd, J = 8.0, 2.3, 1.2 Hz, 1H), 3.82 (s, 2H). C1-17 ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 10.30 (s, 1H), 8.19 (tq, J = 5.1, 2.9, 2.2 Hz, 1H), 8.05 - 7.83 (m, 5H), 7.80 - 7.71 (m , 1H), 7.65 (t, J = 7.8 Hz, 1H), 7.57 (d, J = 8.6 Hz, 2H), 7.44 (d, J = 8.5 Hz, 2H), 4.07 (s, 2H). C1-18 ¹ H NMR (300 MHz, Chloroform-d) δ 15.47 (s, 1H), 8.27 - 8.18 (m, 1H), 8.03 - 7.95 (m, 2H), 7.89 (dd, J = 7.4, 1.6 Hz, 1H) , 7.84 - 7.70 (m, 2H), 7.47 - 7.39 (m, 2H), 7.30 (s, 1H), 7.20 - 7.10 (m, 2H), 7.05 (ddd, J = 8.0, 2.0, 1.1 Hz, 1H) , 6.96 - 6.89 (m, 1H), 3.79 (s, 2H). C1-19 ¹ H NMR (300 MHz, Chloroform-d) δ 15.48 (s, 1H), 8.22 (dd, J = 7.3, 1.6 Hz, 1H), 8.05 - 7.94 (m, 2H), 7.88 (dd, J = 7.2, 1.7 Hz, 1H), 7.83 - 7.68 (m, 2H), 7.49 - 7.38 (m, 2H), 7.35 (s, 1H), 7.23 (d, J = 8.2 Hz, 1H), 7.12 (s, 1H), 6.94 (dt, J = 9.2, 2.6 Hz, 2H), 3.80 (s, 2H). C1-20 ¹ H NMR (300 MHz, Chloroform-d) δ 15.47 (s, 1H), 8.28 - 8.21 (m, 1H), 8.05 - 7.96 (m, 2H), 7.92 (dd, J = 7.4, 1.6 Hz, 1H) , 7.80 (pd, J = 7.5, 1.6 Hz, 2H), 7.51 (t, J = 6.7 Hz, 3H), 7.46 - 7.38 (m, 2H), 7.26 (d, J = 8.4 Hz, 2H), 3.84 ( s, 2H). C1-21 ¹ H NMR (300 MHz, Chloroform-d) δ 15.55 (s, 1H), 8.17 - 8.09 (m, 1H), 7.96 (s, 1H), 7.85 (dd, J = 7.6, 1.5 Hz, 1H), 7.72 (dtd, J = 21.0, 7.6, 1.4 Hz, 2H), 7.48 (d, J = 1.4 Hz, 1H), 7.24 - 7.17 (m, 2H), 7.16 - 7.06 (m, 1H), 3.98 (d, J = 64.9 Hz, 2H), 3.22 (h, J = 6.9 Hz, 1H), 1.34 (s, 3H), 1.21 (s, 3H). C1-22 ¹ H NMR (300 MHz, Chloroform-d) δ 15.55 (s, 1H), 8.19 - 8.08 (m, 1H), 8.02 (s, 1H), 7.85 (dd, J = 7.6, 1.5 Hz, 1H), 7.79 - 7.59 (m, 2H), 7.41 (s, 1H), 7.32 (t, J = 8.1 Hz, 1H), 7.23 (d, J = 7.8 Hz, 1H), 6.99 (d, J = 8.2 Hz, 1H) , 3.99 (d, J = 57.4 Hz, 2H), 3.23 (p, J = 6.8 Hz, 1H), 1.34 (s, 3H), 1.23 (d, J = 11.9 Hz, 3H). C1-23 ¹ H NMR (300 MHz, Chloroform-d) δ 15.55 (s, 1H), 8.17 - 8.07 (m, 2H), 7.86 (dd, J = 7.4, 1.5 Hz, 1H), 7.72 (dtd, J = 20.1, 7.6, 1.4 Hz, 2H), 7.62 - 7.49 (m, 5H), 4.00 (d, J = 80.6 Hz, 2H), 3.23 (p, J = 6.8 Hz, 1H), 1.34 (s, 3H), 1.21 ( s, 3H). C1-24 ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 10.18 (s, 1H), 8.26 (dd, J = 8.7, 5.0 Hz, 1H), 8.13 - 7.98 (m, 2H), 7.87 (dd, J = 7.5) , 2.6 Hz, 1H), 7.79 (td, J = 8.7, 2.6 Hz, 1H), 7.70 (t, J = 8.9 Hz, 1H), 7.32 - 7.11 (m, 2H), 7.01 (dd, J = 8.1, 1.9 Hz, 1H), 6.83 (td, J = 8.5, 2.6 Hz, 1H), 4.07 (s, 2H). C1-25 ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.28 (s, 1H), 8.25 (t, J = 7.1 Hz, 1H), 8.13 - 7.94 (m, 2H), 7.86 (d, J = 7.4 Hz, 1H), 7.78 (t, J = 8.9 Hz, 1H), 7.68 (t, J = 8.8 Hz, 1H), 7.40 - 7.28 (m, 2H), 7.23 (d, J = 8.3 Hz, 1H), 6.99 ( d, J = 8.2 Hz, 1H), 4.06 (s, 2H). C1-26 ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.38 (s, 1H), 8.25 (t, J = 7.2 Hz, 1H), 8.13 - 7.95 (m, 2H), 7.86 (d, J = 7.4 Hz, 1H), 7.79 (t, J = 8.8 Hz, 1H), 7.69 (t, J = 9.3 Hz, 1H), 7.59 (d, J = 8.4 Hz, 2H), 7.47 (d, J = 8.4 Hz, 2H) , 4.10 (s, 2H). C1-27 ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.21 (s, 1H), 8.26 (d, J = 7.7 Hz, 1H), 8.12 - 7.95 (m, 2H), 7.86 (d, J = 7.4 Hz, 1H), 7.78 (t, J = 8.9 Hz, 1H), 7.68 (t, J = 9.2 Hz, 1H), 7.40 (s, 1H), 7.25 (t, J = 8.1 Hz, 1H), 7.15 (d, J = 8.4 Hz, 1H), 7.06 (d, J = 8.1 Hz, 1H), 4.06 (s, 2H). C1-28 ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 16.01 (s, 1H), 10.22 (s, 1H), 8.06 (d, J = 8.2 Hz, 2H), 7.94 (d, J = 8.9 Hz, 1H) , 7.60 (dd, J = 13.6, 8.4 Hz, 3H), 7.47 (d, J = 8.6 Hz, 2H), 7.30 (d, J = 9.3 Hz, 1H), 7.17 (s, 1H), 3.94 (s, 2H), 3.54 (s, 4H), 1.68 - 1.52 (m, 6H). C1-29 ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 16.14 (s, 1H), 10.21 (s, 1H), 8.04 (t, J = 9.3 Hz, 2H), 7.92 (d, J = 8.8 Hz, 1H) , 7.60 (dd, J = 12.7, 6.0 Hz, 4H), 7.48 (d, J = 8.5 Hz, 2H), 6.92 (d, J = 10.2 Hz, 2H), 3.94 (s, 2H), 2.86 (t, J = 3.3 Hz, 3H). C1-30 ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 10.28 (s, 1H), 9.96 (s, 1H), 8.06 (dd, J = 16.1, 8.0 Hz, 3H), 7.65 (t, J = 8.9 Hz, 1H), 7.59 (d, J = 8.5 Hz, 2H), 7.48 (d, J = 8.5 Hz, 2H), 7.42 (dd, J = 9.1, 2.5 Hz, 1H), 7.37 (d, J = 2.6 Hz, 1H), 4.32 (d, J = 13.0 Hz, 2H), 4.01 (s, 2H), 3.56 (d, J = 12.3 Hz, 2H), 3.29 (s, 2H), 3.16 (s, 2H), 2.88 ( s, 3H). C1-31 ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 10.28 (s, 1H), 8.22 - 8.12 (m, 1H), 8.07 (d, J = 1.9 Hz, 1H), 7.96 - 7.84 (m, 5H), 7.41 - 7.28 (m, 2H), 7.20 (d, J = 8.2 Hz, 1H), 7.04 - 6.92 (m, 1H), 4.03 (s, 2H). C1-32 ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 10.34 (s, 1H), 8.21 - 8.15 (m, 1H), 8.08 (d, J = 1.9 Hz, 1H), 7.96 - 7.87 (m, 5H), 7.57 (d, J = 8.6 Hz, 2H), 7.45 (d, J = 8.5 Hz, 2H), 4.07 (s, 2H). C1-33 ¹ H NMR (400 MHz, CDCl ₃ ) δ 15.36 (s, 1H), 8.25 - 8.21 (m, 1H), 8.07 (d, J = 2.0 Hz, 1H), 7.95 - 7.88 (m, 2H), 7.82 - 7.73 (m, 2H), 7.50 (d, J = 8.3 Hz, 1H), 7.30 (s, 1H), 7.20 (t, J = 2.1 Hz, 1H), 7.15 (t, J = 8.0 Hz, 1H), 7.05 (ddd, J = 8.0, 2.0, 1.0 Hz, 1H), 6.95 (ddd, J = 8.1, 2.2, 1.0 Hz, 1H), 3.81 (s, 2H). C1-34 ¹ H NMR (400 MHz, CDCl ₃ ) δ 15.36 (s, 1H), 8.31 (dt, J = 8.0, 1.4 Hz, 1H), 8.26 (t, J = 1.7 Hz, 1H), 8.25 - 8.22 (m, 1H), 7.91 - 7.88 (m, 1H), 7.79 (td, J = 7.6, 1.5 Hz, 1H), 7.76 - 7.72 (m, 1H), 7.70 (dt, J = 7.9, 1.4 Hz, 1H), 7.54 (td, J = 7.9, 0.7 Hz, 1H), 7.41 (s, 1H), 7.24 (d, J = 8.2 Hz, 1H), 7.12 (s, 1H), 6.98 (dddd, J = 23.6, 8.3, 2.3 , 1.1 Hz, 2H), 3.79 (s, 2H). C1-35 ¹ H NMR (400 MHz, CDCl ₃ ) δ 15.36 (s, 1H), 8.32 (ddd, J = 7.9, 1.7, 1.2 Hz, 1H), 8.27 - 8.20 (m, 2H), 7.89 (dd, J = 7.7) , 1.4 Hz, 1H), 7.83 - 7.71 (m, 3H), 7.55 (td, J = 7.8, 0.6 Hz, 1H), 7.40 (s, 1H), 7.18 - 7.12 (m, 2H), 7.06 (ddd, J = 8.0, 2.0, 1.0 Hz, 1H), 6.97 (ddd, J = 8.1, 2.1, 1.1 Hz, 1H), 3.78 (s, 2H). C1-36 ¹ H NMR (400 MHz, CDCl ₃ ) δ 15.34 (s, 1H), 8.32 (ddd, J = 7.9, 1.8, 1.2 Hz, 1H), 8.30 - 8.18 (m, 2H), 7.93 - 7.86 (m, 1H) ), 7.82 - 7.73 (m, 2H), 7.70 (dt, J = 7.9, 1.4 Hz, 1H), 7.53 (td, J = 7.9, 0.6 Hz, 1H), 7.51 - 7.44 (m, 3H), 7.28 - 7.26 (m, 1H), 7.25 (s, 1H), 3.81 (s, 2H). C1-37 ¹ H NMR (400 MHz, Acetone) δ 15.28 (s, 1H), 8.40 - 8.30 (m, 3H), 8.06 (dt, J = 7.8, 1.4 Hz, 1H), 8.02 - 7.97 (m, 2H), 7.93 - 7.90 (m, 1H), 7.84 - 7.78 (m, 2H), 7.71 (ddd, J = 8.7, 4.7, 2.2 Hz, 1H), 7.33 (t, J = 8.8 Hz, 1H), 4.89 (s, 2H) ). C1-38 ¹ H NMR (400 MHz, Acetone) δ 15.47 (s, 1H), 9.29 (s, 1H), 8.30 - 8.21 (m, 3H), 8.04 - 7.99 (m, 2H), 7.96 - 7.89 (m, 3H) , 7.40 (s, 1H), 7.32 (t, J = 8.2 Hz, 1H), 7.24 - 7.19 (m, 1H), 6.96 (ddt, J = 8.2, 2.3, 1.1 Hz, 1H), 4.04 (s, 2H) ). C1-39 ¹ H NMR (400 MHz, Acetone) δ 15.47 (s, 1H), 9.22 (s, 1H), 8.28 - 8.22 (m, 3H), 8.03 - 8.00 (m, 2H), 7.96 - 7.90 (m, 3H) , 7.46 (t, J = 2.0 Hz, 1H), 7.20 (t, J = 8.0 Hz, 1H), 7.14 (dt, J = 8.5, 1.3 Hz, 1H), 7.03 (ddd, J = 7.8, 2.1, 1.2) Hz, 1H), 4.03 (s, 2H). C1-40 ¹ H NMR (400 MHz, Acetone) δ 9.41 (s, 1H), 8.30 - 8.22 (m, 3H), 8.01 (d, J = 8.3 Hz, 2H), 7.96 (dd, J = 5.8, 3.3 Hz, 2H) ), 7.92 - 7.89 (m, 1H), 7.53 (d, J = 9.0 Hz, 2H), 7.49 (d, J = 8.9 Hz, 2H), 4.08 (s, 2H). C1-41 ¹ H NMR (400 MHz, Acetone) δ 15.32 (s, 1H), 8.35 - 8.29 (m, 1H), 8.23 - 8.19 (m, 2H), 8.02 - 7.97 (m, 4H), 7.92 - 7.88 (m, 1H), 7.78 (dd, J = 7.1, 2.2 Hz, 1H), 7.70 (ddd, J = 8.7, 4.7, 2.2 Hz, 1H), 7.33 (t, J = 8.8 Hz, 1H), 4.85 (s, 2H) ). C1-42 ¹ H NMR (400 MHz, Acetone) δ 15.98 (s, 1H), 9.27 (s, 1H), 8.26 - 8.21 (m, 1H), 8.19 - 8.12 (m, 2H), 7.93 - 7.86 (m, 3H) , 7.39 (d, J = 2.8 Hz, 1H), 7.30 (t, J = 8.2 Hz, 1H), 7.22 (ddd, J = 8.3, 2.0, 1.0 Hz, 1H), 7.16 - 7.12 (m, 2H), 6.95 (ddt, J = 8.2, 2.3, 1.2 Hz, 1H), 4.11 (s, 2H), 3.94 (s, 3H). C1-43 ¹ H NMR (400 MHz, Acetone) δ 15.97 (s, 1H), 9.17 (s, 1H), 8.25 - 8.23 (m, 1H), 8.18 - 8.15 (m, 2H), 7.92 - 7.88 (m, 3H) , 7.46 (t, J = 2.0 Hz, 1H), 7.22 - 7.15 (m, 2H), 7.15 - 7.09 (m, 3H), 7.01 (ddd, J = 7.6, 2.1, 1.3 Hz, 1H), 4.09 (s) , 2H), 3.95 (s, 3H). C1-44 ¹ H NMR (400 MHz, Acetone) δ 15.97 (s, 1H), 9.35 (s, 1H), 8.28 - 8.21 (m, 1H), 8.20 - 8.13 (m, 2H), 7.95 - 7.86 (m, 3H) , 7.54 - 7.47 (m, 4H), 7.17 - 7.11 (m, 2H), 4.12 (s, 2H), 3.94 (s, 3H). C1-45 ¹ H NMR (400 MHz, Acetone) δ 15.85 (s, 1H), 8.31 - 8.25 (m, 1H), 8.15 - 8.10 (m, 2H), 7.95 (ddd, J = 6.0, 2.9, 1.9 Hz, 2H) , 7.89 - 7.86 (m, 1H), 7.77 (dd, J = 7.2, 2.2 Hz, 1H), 7.71 (ddd, J = 8.7, 4.6, 2.2 Hz, 1H), 7.32 (t, J = 8.8 Hz, 1H) ), 7.08 (d, J = 8.9 Hz, 2H), 4.84 (s, 2H), 3.94 (s, 3H). C1-46 ¹ H NMR (400 MHz, DMSO) δ 14.40 (s, 1H), 8.23 - 8.16 (m, 1H), 8.05 (s, 1H), 8.00 - 7.79 (m, 6H), 7.76 - 7.72 (m, 1H) , 7.45 (t, J = 8.9 Hz, 1H), 4.86 (s, 2H). C1-47 ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 10.17 (s, 1H), 8.24 - 8.17 (m, 1H), 8.09 - 7.95 (m, 4H), 7.90 (s, 1H), 7.67 (dd, J = 17.8, 8.8 Hz, 2H), 7.46 (d, J = 13.5 Hz, 1H), 7.43 - 7.33 (m, 2H), 7.00 (d, J = 7.8 Hz, 1H), 5.56 (d, J = 13.5 Hz) , 1H). C1-48 C1-49 ¹ H NMR (500 MHz, Chloroform-d) δ 8.77 (s, 1H), 7.98 - 7.92 (m, 3H), 7.84 - 7.70 (m, 3H), 7.52 - 7.45 (m, 3H), 7.38 - 7.28 ( m, 2H), 7.00 - 6.94 (m, 1H), 4.09 (s, 2H), 3.56 (s, 3H). C1-50 ¹ H NMR (300 MHz, Chloroform-d) δ 8.27 (s, 1H), 8.15 (dd, J = 7.9, 1.3 Hz, 1H), 7.82 (dd, J = 7.6, 1.4 Hz, 1H), 7.71 (td) , J = 7.7, 1.4 Hz, 1H), 7.64 - 7.43 (m, 5H), 4.03 (s, 2H), 1.46 (s, 9H). C1-51 ¹ H NMR (400 MHz, Chloroform-d) δ 8.13 - 8.04 (m, 2H), 7.77 (dd, J = 7.7, 1.3 Hz, 1H), 7.65 (td, J = 7.7, 1.3 Hz, 1H), 7.55 (td, J = 7.7, 1.3 Hz, 1H), 7.31 (d, J = 2.4 Hz, 1H), 7.28 (d, J = 8.2 Hz, 1H), 7.15 (ddd, J = 8.2, 2.1, 1.0 Hz, 1H), 6.96 (ddt, J = 8.2, 2.2, 1.1 Hz, 1H), 4.00 (s, 2H), 1.44 (s, 9H).

Experimental Example 1. Evaluation of anti-COVID-19 activity

1-1. Cell line and virus preparation

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

SARS-CoV-2 Korean isolate (βCoV/KOR/KCDC03/2020, Genbank accession no. KT029139.1) was provided by the National Institutes of Health, Korea Centers for Disease Control and Prevention, Jeon et al., 2020 DOI: 10.1128/AAC.00819-20 Proliferated in Vero cells according to the method presented in All experiments using SARS-CoV-2 were performed at the Pasteur Institute in Korea, which complied with the National Institutes of Health's enhanced Biosafety Level 3 (BL-3) containment procedures approved by the Korea Centers for Disease Control and Prevention.

1-2. reagent preparation

Chloroquine diphosphate (CQ; C6628), lopinavir (LPV; GP6351)), and remdesivir were purchased from SelleckChem (Houston, TX) and Glentham Life Science (UK), respectively. The anti-SARS-CoV-2 spike antibody used as the primary antibody was obtained from Sino Biological Inc. (Beijing, China). The secondary antibody Alexa Fluor 488 goat anti-rabbit IgG and the nuclear chromosome Hoechst 33342 were purchased from MolecularProbes/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).

1-3. Image-based assays using immunofluorescence assays

Since cells infected with SARS-CoV-2 express the viral protein, it can be measured using an antibody that specifically binds to the viral protein. In the present invention, cells were stained using an antibody binding to the nucleocapsid protein of SARS-CoV-2, and infected cells were imaged through a microscope. The infection rate (number of cells expressing SARS-CoV-2 spike protein/total number of cells) was measured with the internally developed Image Mining 3.0 (IM 3.0) plugin. In order to compare the antiviral effects of small molecule compounds, dimethyl sulfoxide (DMSO)-treated infected cells were used as a negative control, and three compounds (CQ, LPV, Remdesivir) with known antiviral activity against SARS-CoV-2. was used as a positive control to optimize the image-based assay.

1-4. Small molecule compound library

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

1-5. Image-based small molecule compound screening

Vero cells at 1.2×10 ⁴ cells per well in Opti-PRO™ SFM with 4 mM L-Glutamine and 1× 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 prior to viral infection using an automated liquid handling system (Apricot Designs, Covina, CA). The final concentration of the test compound was 2.5-28.2 μM, and the concentration of DMSO was maintained at 0.5%. The compound-treated group was transferred to a BL-3 containment chamber and then infected with SARS-CoV-2 at an MOI of 0.0125.

The infection was fixed using PFA (final PAF concentration = 4%) 24 hours after infection. After treatment with anti-SARS-CoV-2 nucleocapsid antibody, fixed cells were stained with Alexa Fluor 488 goat anti-rabbit IgG and Hoechst 33342. Infected cells were imaged on a fluorescence imaging system (Perkin Elmer Operetta, 20×, Waltham, Mass.) at 20× magnification after fixation and staining. The SARS-CoV-2 infection rate of cells treated with the small molecule compound was converted into a negative control (0% infection inhibition rate) and a positive control group (100% infection inhibition rate) on each plate, resulting in more than 90% inhibitory effect low molecular weight compounds have been identified.

1-6. Concentration-response curve experiment of active compound

The inhibitory effect of virus infection according to the concentration of the compound can be known through the concentration-response curve experiment. The highest concentration of the test compound is 5 mM, diluted two-fold with DMSO, and serially diluted up to 10 steps. This is treated with the cells prepared in the same way as in 1-5 above. The maximum concentration of the final concentration of the test 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 infected with SARS-CoV-2 at an MOI of 0.0125. 24 hours after infection, the infection rate was imaged and converted in the same manner as in 1-5 above. The 50% virus inhibitory concentration (Inhibitory concentration 50; IC ₅₀ ), 50% cytotoxicity concentration (CC ₅₀ ), and the ratio between drug and cytotoxicity (selectivity index; SI) of the compound were calculated by the concentration-response curve experiment. . The results are shown in Table 2 below.

compound IC ₅₀ (μM) CC ₅₀ SI C1-1 NOI >25 1.0 C1-2 NOI >25 1.0 C1-3 11.50 >25 2.1 C1-4 14.80 >25 1.4 C1-5 NOI >25 1.0 C1-6 15.30 >25 1.6 C1-7 8.90 >25 2.7 C1-8 5.30 >25 4.7 C1-9 5.30 >25 4.7 C1-10 9.20 >25 2.8 C1-11 2.50 >25 11.1 C1-12 12.60 >25 1.8 C1-13 10.10 >25 2.3 C1-14 7.00 >25 3.2 C1-15 8.90 >25 2.6 C1-16 11.90 >25 2.1 C1-17 4.10 >25 5.8 C1-18 6.60 >25 3.7 C1-19 8.50 >25 2.9 C1-20 4.00 >25 6.0 C1-21 14.00 >25 1.3 C1-22 10.80 >25 1.9 C1-23 7.30 >25 3.0 C1-24 4.00 >25 6.3 C1-25 3.10 >25 8.9 C1-26 0.88 >25 30.7 C1-27 2.20 >25 12.1 C1-28 2.10 >25 5.6 C1-29 13.80 >25 1.3 C1-30 14.00 >25 1.6 C1-31 6.60 >25 4.0 C1-32 2.80 >25 8.5 C1-33 4.20 >25 5.8 C1-34 14.30 >25 1.6 C1-35 >25 >25 1.0 C1-36 10.70 >25 1.5 C1-37 >25 >25 1.0 C1-38 14.30 >25 1.6 C1-39 >25 >25 1.0 C1-40 9.30 >25 1.4 C1-41 >25 >25 1.0 C1-42 11.60 >25 2.0 C1-43 >25 >25 1.0 C1-44 8.60 >25 2.5 C1-45 12.40 >25 1.3 C1-46 4.80 >25 5.2 C1-47 6.60 >25 3.6 C1-48 12.70 >25 1.9 C1-49 5.30 >25 4.7 C1-50 1.90 >25 13.1 C1-51 3.30 >25 7.6 chloroquine 7.28 >150 20.61 lopinavir 9.12 >50 5.48 remdesivir 11.41 >25 2.19

As shown in Table 2, the compound according to one aspect of the present invention has excellent antiviral inhibitory activity against SARS-CoV-2, so it can be seen that it can be used as a treatment for COVID-19 (coronavirus infection-19). have. It can also be used as a treatment for severe acute respiratory syndrome (SARS) or Middle East respiratory syndrome (MERS).

As described above, the present invention has been described by way of example, and various modifications will be possible without departing from the essential characteristics of the present invention by those of ordinary skill in the art to which the present invention pertains. Accordingly, the embodiments disclosed herein are for illustrative purposes rather than limiting the present invention, and the spirit and scope of the present invention are not limited by these embodiments. The protection scope of the present invention should be construed by the following claims, and all technologies within the scope equivalent thereto should be construed as being included in the scope of the present invention.

Claims (12)

delete delete delete delete delete A compound characterized in that it is represented by the following formula (6)
[Formula 6]

{In Formula 6,
1) each R ₄ is the same or different and is hydrogen; halogen; O, N, S containing at least one hetero atom C ₂ ~ C ₂₄ A heterocyclic group; or -NR ^a R ^b ; and a is an integer from 0 to 4,
2) R ₅ and R ₈ are each the same or different, and each independently hydrogen; halogen; cyano group; C ₁ ~ C ₁₀ Alkyl group; C ₂ ~ C ₁₀ Alkenyl group; C ₂ ~ C ₁₀ Alkynyl group; And C ₁ ~ C ₁₀ An alkoxy group; is selected from the group consisting of, b is an integer of 0 to 5 independently of each other.}
delete An antiviral pharmaceutical composition comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient
[Formula 1]

{In Formula 1,
1) X is O or NOCH ₃ ,
2) R ₁ is a C ₁ ~ C ₁₀ alkyl group; C ₂ ~ C ₁₀ Alkenyl group; C ₂ ~ C ₁₀ Alkynyl group; Or C ₆ ~ C ₂₄ Aryl group;
3) R ₂ is a hydroxyl group; Or C ₁ ~ C ₁₀ Alkoxy group;
4) L is a C ₁ ~ C ₃ alkylene group; Or C ₂ ~ C ₃ An alkenylene group;
5) R ₃ is a C ₆ ~ C ₂₄ aryl group; -NH-C ₆ ~ C ₂₄ Aryl group; Or O, N, S containing at least one hetero atom C ₂ ~ C ₁₂ A heterocyclic group;
6) each R ₄ is the same or different and is hydrogen; halogen; O, N, S containing at least one hetero atom C ₂ ~ C ₂₄ A heterocyclic group; or -NR ^a R ^b ;
7) wherein R ^a and R ^b are each independently hydrogen; Or C ₁ ~ C ₁₀ Alkyl group;
8) a is an integer from 0 to 4,
9) Here, the alkyl group, the alkenyl group, the alkynyl group, the alkoxy group, the aryl group and the heterocyclic group are each deuterium; halogen; hydroxyl group; cyano group; nitro group; C ₁ ~ C ₆ Alkyl group; C ₂ ~ C ₆ An alkenyl group; C ₂ ~ C ₆ Alkynyl group; C ₁ ~ C ₆ Alkoxy group; C ₁ ~ C ₆ Alkyl group substituted with fluorine; C ₁ ~ C ₆ Alkoxy group substituted with fluorine; C ₆ ~ C ₁₂ Aryl group; And O, N, S including at least one heteroatom C ₂ ~ C ₁₀ A heterocyclic group; may be further substituted with one or more substituents selected from the group consisting of.}
The pharmaceutical composition according to claim 8, wherein the virus is a coronavirus.
The pharmaceutical composition according to claim 9, wherein the coronavirus is SARS coronavirus (SARS-CoV), MERS coronavirus (MERS-CoV), or SARS coronavirus 2 (SARS-CoV2).
The pharmaceutical composition according to claim 8, further comprising at least one pharmaceutically acceptable carrier.
The pharmaceutical composition according to claim 8, further comprising at least one antiviral agent.