KR102010274B1

KR102010274B1 - New benzamide compound or pharmaceutically acceptable salt thereof having inhibitory activity on Hsp90 and medical use thereof

Info

Publication number: KR102010274B1
Application number: KR1020160011622A
Authority: KR
Inventors: 서영호
Original assignee: 계명대학교 산학협력단
Priority date: 2016-01-29
Filing date: 2016-01-29
Publication date: 2019-08-13
Also published as: KR20170090810A

Abstract

The present invention relates to a composition comprising a novel benzamide compound or a novel benzamide compound having Hsp90 inhibitory activity, and the compound can effectively inhibit Hsp90, and thus is selected from the group consisting of cancer diseases, degenerative neurological diseases and viral infections. It can be usefully used as a pharmaceutical composition for preventing or treating any one of Hsp90 mediated diseases or as a nutraceutical for preventing or improving.

Description

New benzamide compound or pharmaceutically acceptable salt thereof and its medical use {New benzamide compound or pharmaceutically acceptable salt having having inhibitory activity on Hsp90 and medical use}

To a novel benzamide compound having a Hsp90 inhibitory activity, or a pharmaceutically acceptable salt thereof, and a medical use thereof.

Hsp90 protein is one of the most common chaperones in eukaryotic cells, and is responsible for stabilization and activity regulation of various proteins involved in cell growth differentiation and survival. Hsp90 matrix proteins, called client proteins, contain about 50 cancer-causing proteins. When Hsp90 activity is inhibited, Hsp90 client proteins are degraded by proteasomes.

Therefore, Hsp90 activity inhibitors have a great interest as an anticancer agent that can be applied to a wide range of cancers because they can have the effect of simultaneously reducing various cancer-causing proteins. In particular, Hsp90 has been reported to be effective in treating cancer with resistance because it simultaneously reduces various cancer-causing proteins.

In addition, some Hsp90 client proteins are known to cause degenerative neurological diseases, and therefore, it has been reported that Hsp90 inhibitors may be used as therapeutic agents for neurodegenerative diseases.

Hsp90 inhibitors began with the development of the natural geldanamycin (GA). GA was found to induce degradation of Src, a client protein, through the inhibition of Hsp90 in 1994. Since then, inhibitors targeting Hsp90 have been actively developed. However, GA has strong anticancer effects but has problems with liver toxicity, solubility and stability. In order to compensate for this, GA derivatives Tanespinmycin (17-AAG), alvespimycin (17-DMAG), retaspimycin (GA) was developed, but the problem was not solved by the structural properties of GA. Subsequently, studies of Hsp90 inhibitors of various structures have been studied in the clinical stage, but since the FDA-approved drug has not been developed yet, it is necessary to develop a compound with new and stronger efficacy.

1. Korean Patent Publication No. 10-2007-0038565.

Targeting HSP90 for cancer therapy; British Journal of Cancer (2009) 100, 1523-1529 2. Targeting HSP90 to Halt Neurodegeneration; Chem Biol. 2006 Feb; 13 (2): 115-6 3.Crystal Structure of an Hsp90-Geldanamycin Complex: Targeting of a Protein Chaperone by an Antitumor Agent; Cell, Vol. 89, 239-250, April 18, 1997

Accordingly, an object of the present invention is to provide a novel benzamide compound.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating Hsp90 mediated diseases, which comprises a novel benzamide compound as an active ingredient.

In another aspect, the present invention is to provide a health functional food for preventing or improving Hsp90 mediated diseases comprising a novel benzamide compound as an active ingredient.

In order to achieve the above object, the present invention provides a benzamide compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof:

[Formula 1]

In Chemical Formula 1,

R ₁ is halogen or C ₁ -C 4 alkyl,

R ₂ is

or

ego,

R ₃ and R ₄ may be the same or different, and each one selected from the group consisting of H, C 1 -C 4 alkyl, and C 1 -C 4 alkoxy,

R ₅ and R ₆ may be the same or different, and each one selected from the group consisting of H, C 1 -C 4 alkyl and C 1 -C 4 alkoxy, or R ₅ and R ₆ are connected to each other to form a 5- or 6-ring ring; Forming,

R ₇ is any one selected from the group consisting of H, C 1 -C 4 alkyl and C 1 -C 4 alkoxy,

R ₈ is H or halogen,

R ₉ is any one selected from the group consisting of C1-C4 alkyl, C1-C4 alkoxy and benzyl groups.

In order to achieve the above another object, the present invention provides a pharmaceutical composition for the prevention or treatment of Hsp90-mediated disease comprising a benzamide compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

In order to achieve the above another object, the present invention provides a health functional food for preventing or ameliorating Hsp90-mediated disease comprising a benzamide compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

1 is 2,4-dihydroxy-5-isopropyl-N-methyl-N- (4- (propylcarbamoyl) benzyl) benzamide [2,4-Dihydroxy-5 in a benzamide compound according to the present invention; -isopropyl-N-methyl-N- (4- (propylcarbamoyl) benzyl) benzamide] confirmed the inhibitory effect of Hsp90.

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail.

The inventors of the present invention, while studying a compound that exhibits a heat shock protein 90 (Heat Shock Protein 90) inhibitory effect, by synthesizing the compound represented by the formula (1) and confirmed its Hsp90 inhibitory effect was completed the present invention.

Accordingly, the present invention provides a benzamide compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof:

[Formula 1]

In Chemical Formula 1,

R ₁ is halogen or C ₁ -C 4 alkyl,

R ₂ is

or

ego,

R ₈ is H or halogen,

In the benzamide compound, in Formula 1, R ₁ is halogen or C ₁ -C 4 alkyl, and R ₂ is

R ₃ is any one selected from the group consisting of H, C 1 -C 4 alkyl and C 1 -C 4 alkoxy, and R ₉ may be any one selected from the group consisting of C 1 -C 4 alkyl, C 1 -C 4 alkoxy and benzyl groups.

Benzamide compound represented by the formula (1) is N-benzyl-5-chloro-2,4-dihydroxybenzamide [N-Benzyl-5-chloro-2,4-dihydroxybenzamide], N-benzyl-5-chloro -2,4-dihydroxy-N-methylbenzamide [N-Benzyl-5-chloro-2,4-dihydroxy-N-methylbenzamide], 5-chloro-2,4-dihydroxy-N- (4 -Methoxybenzyl) -N-methylbenzamide [5-chloro-2,4-dihydroxy-N- (4-methoxybenzyl) -N-methylbenzamide], 5-chloro-N- (3,4-dimethoxybenzyl) -2,4-dihydroxy-N-methylbenzamide [5-chloro-N- (3,4-dimethoxybenzyl) -2,4-dihydroxy-N-methylbenzamide], N- (benzo [d] [[1 , 3] dioxol-5-ylmethyl) -5-chloro-2,4-dihydroxy-N-methylbenzamide [N- (benzo [d] [1,3] dioxol-5-ylmethyl) -5 -chloro-2,4-dihydroxy-N-methylbenzamide], 5-chloro-2,4-dihydroxy-N-methyl-N- (3,4,5-trimethoxybenzyl) benzamide [5-chloro -2,4-dihydroxy-N-methyl-N- (3,4,5-trimethoxybenzyl) benzamide], 5-chloro-N- (2-chloro-3,4,5-trimethoxybenzyl) -2, 4-dihydroxy-N-methylbenzami De [5-chloro-N- (2-chloro-3,4,5-trimethoxybenzyl) -2,4-dihydroxy-N-methylbenzamide], N- (2-bromo-3,4,5-trimethoxy Benzyl) -5-chloro-2,4-dihydroxy-N-methylbenzamide [N- (2-bromo-3,4,5-trimethoxybenzyl) -5-chloro-2,4-dihydroxy-N-methylbenzamide ], 5-chloro-2,4-dihydroxy-N-methyl-N- (3-methylbenzyl) benzamide [5-chloro-2,4-dihydroxy-N-methyl-N- (3-methylbenzyl) benzamide], 5-chloro-2,4-dihydroxy-N-methyl-N- (2-methylbenzyl) benzamide [5-chloro-2,4-dihydroxy-N-methyl-N- (2-methylbenzyl ) benzamide], 5-chloro-2,4-dihydroxy-N-methyl-N- (4-methylbenzyl) benzamide [5-chloro-2,4-dihydroxy-N-methyl-N- (4- methylbenzyl) benzamide], 5-chloro-2,4-dihydroxy-N-methyl-N- (4- (methylcarbamoyl) benzyl) benzamide [5-chloro-2,4-dihydroxy-N-methyl- N- (4- (methylcarbamoyl) benzyl) benzamide], 5-chloro-N- (4- (ethylcarbamoyl) benzyl) -2,4-dihydroxy-N-methylbenzamide [5-chloro-N- (4- (ethylcarbamoyl) benzyl) -2,4-dihydroxy-N-methylbenzamide], 5-cle Loro-2,4-dihydroxy-N-methyl-N- (4- (propylcarbamoyl) benzyl) benzamide [5-chloro-2,4-dihydroxy-N-methyl-N- (4- (propylcarbamoyl ) benzyl) benzamide], 2,4-dihydroxy-5-isopropyl-N-methyl-N- (4- (methylcarbamoyl) benzyl) benzamide [2,4-dihydroxy-5-isopropyl-N- methyl-N- (4- (methylcarbamoyl) benzyl) benzamide], N- (4- (ethylcarbamoyl) benzyl) -2,4-dihydroxy-5-isopropyl-N-methylbenzamide [N- ( 4- (ethylcarbamoyl) benzyl) -2,4-dihydroxy-5-isopropyl-N-methylbenzamide], 2,4-dihydroxy-5-isopropyl-N-methyl-N- (4- (propylcarbamoyl) Benzyl) benzamide [2,4-dihydroxy-5-isopropyl-N-methyl-N- (4- (propylcarbamoyl) benzyl) benzamide] and N-benzyl-2,4-dihydroxy-5-isopropyl-N -Methyl benzamide [N-benzyl-2,4-dihydroxy-5-isopropyl-N-methylbenzamide] may be any one selected from the group consisting of, preferably 2,4-dihydroxy-5-isopropyl -N-methyl-N- (4- (propylcarbamoyl) benzyl) benzamide.

In addition, the present invention provides a pharmaceutical composition for preventing or treating a heat shock protein 90 (Hsp90) mediated disease comprising a benzamide compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient:

[Formula 1]

In Chemical Formula 1,

R ₁ is halogen or C ₁ -C 4 alkyl,

R ₂ is

or

ego,

R ₈ is H or halogen,

The heat shock protein 90 mediated disease is any one or more diseases selected from the group consisting of cancer diseases, neurodegenerative diseases and viral infections.

The cancer diseases include non-small cell lung cancer, breast cancer, ovarian cancer, uterine cancer, pancreatic cancer, lung cancer, stomach cancer, liver cancer, colon cancer, skin cancer, head or neck cancer, brain cancer, laryngeal cancer, prostate cancer, bladder cancer, esophageal cancer, thyroid cancer, kidney cancer, and rectal cancer. , Acute myeloid leukemia, chronic myeloid leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia and hematologic cancer may be any one selected from the group consisting of, but is not limited thereto.

The neurodegenerative disorders include stroke, stroke, memory loss, memory impairment, dementia, forgetfulness, Parkinson's disease, Alzheimer's disease, Pick disease, Creutzfeld-Kacob disease, Huntington's disease, and Lou Gehrig's disease. It may be any one selected, but is not limited thereto.

The pharmaceutical compositions according to the invention may further comprise suitable carriers, excipients or diluents commonly used in the manufacture of pharmaceutical compositions.

Carriers, excipients or diluents usable in the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, Methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like.

The pharmaceutical compositions according to the invention can be used in the form of oral dosage forms, external preparations, suppositories, and sterile injectable solutions, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., respectively, according to conventional methods. Can be.

When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations may include at least one excipient, for example, starch, calcium carbonate, sucrose ( sucrose, lactose, gelatin and the like can be mixed and prepared.

In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. .

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

In addition, the dosage of the pharmaceutical composition according to the present invention may be increased or decreased depending on the route of administration, the severity of the disease, sex, weight, age, and the like. Therefore, the above dosage does not limit the scope of the present invention in any aspect.

The pharmaceutical composition may be administered to mammals such as mice, mice, livestock, humans, and the like by various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrabronchial inhalation, intrauterine dural or intracerebroventricular injection.

The benzamide compound of the present invention can be used in the form of a pharmaceutically acceptable salt, and acid salts formed by the pharmaceutically acceptable free acid are useful as salts. The inorganic acid and organic acid may be used as the free acid, and hydrochloric acid, bromic acid, sulfuric acid, sulfurous acid, phosphoric acid, etc. may be used as the inorganic acid, and citric acid, acetic acid, maleic acid, fumaric acid, gluconic acid, and methanesulfonic acid may be used as the organic acid. , Acetic acid, glyconic acid, succinic acid, tartaric acid, 4-toluenesulfonic acid, galacturonic acid, embonic acid, glutamic acid, citric acid, aspartic acid, and the like can be used. Preferably, hydrochloric acid may be used as the inorganic acid, and methanesulfonic acid may be used as the organic acid.

In addition, the benzamide compounds of the present invention include not only pharmaceutically acceptable salts, but also all salts, hydrates, and solvates that can be prepared by conventional methods.

The addition salt according to the present invention can be prepared by a conventional method, for example, by dissolving the benzamide compound of formula (1) in a water miscible organic solvent such as acetone, methanol, ethanol, or acetonitrile and adding an excess of an organic acid. Or by adding an aqueous acid solution of an inorganic acid and then precipitating or crystallizing. The solvent or excess acid may then be evaporated and dried in this mixture to obtain an addition salt or the precipitated salt may be prepared by suction filtration.

In another aspect, the present invention provides a dietary supplement for the prevention or improvement of heat shock protein 90 (Hsp90) mediated diseases comprising a benzamide compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient :

[Formula 1]

In Chemical Formula 1,

R ₁ is halogen or C ₁ -C 4 alkyl,

R ₂ is

or

ego,

R ₈ is H or halogen,

The dietary supplement may be provided in the form of a powder, granules, tablets, capsules, syrups or beverages. The dietary supplement is used with other foods or food additives in addition to the compound represented by Formula 1 as an active ingredient. It can be suitably used according to the phosphorus method. The mixed amount of the active ingredient can be suitably determined depending on the purpose of use thereof, for example, prophylactic, health or therapeutic treatment.

The benzamide compound of Formula 1 contained in the health functional food can be used in accordance with the effective dose of the pharmaceutical composition, but in the case of prolonged intake for the purpose of health and hygiene or for health control It can be hereinafter, it is obvious that the active ingredient can be used in an amount above the above range because there is no problem in terms of safety.

There are no particular limitations on the types of dietary supplements, for example, meat, sausages, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, drinks, tea , A drink, an alcoholic beverage, and a vitamin complex.

Hereinafter, examples will be described in detail to help understand the present invention. However, the following examples are merely to illustrate the content of the present invention is not limited to the scope of the present invention. The embodiments of the present invention are provided to more completely explain the present invention to those skilled in the art.

< Reference Example 1> Reagent And devices

Reagents are Sigma-aldrich, St. Louis, Mo, USA, acros organics, Thermo Fisher Scientific In., Gell, Belgium, alfa aesar, A Johnson Mattey Company , Karlsruhe, Germany) and Daejung (Daejeong Co., Ltd., Gyeonggi-do, Korea) were used. Compound synthesis was carried out under argon or air, and microwaves (Biotage ^® , Uppsala, Sweden) were optionally used. Extraction, recrystallization, column chromatography and medium pressure liquid column chromatography (MPLC) were used for purification of the product after synthesis. The filler used in column chromatography was silica gel 60 (0.040-0.063 mm), and MPLC used SNAP cartridge (KP-Sil 25g or KP-C18-HS 30g).

¹ H NMR and ¹³ C NMR spectra of each compound were determined using a Bruker spectrospin 400 spectrometer (Bruker co., Billerica, Massachusetts, USA). The solvent used was CDCl ₃ , CD ₃ OD or dimethyl sulfoxide (DMSO) -d ₆ , and the chemical shift (δ) was expressed in ppm, and the peaks were d (doublet) and t (triplet). ), m (multiplet) and dd (doublet of doublet).

A power supply (BioRad co., Hercules, CA), an image analyzer (Fuji, Tokyo, Japan), and a microplate reader (microplate reader, TECAN, Mannedorf, Switerland) were used for the biological activity evaluation process.

< Example 1> Compound Synthesis

Compounds 6a to 6h and 8a to 8c were synthesized in the same manner as in Scheme 1 below.

Scheme 1

One. Amine Synthesis Method I ( 6a-h )

1 equivalent of each aldehyde (aldehyde) and 1.5 equivalents of methylamine (methylamine in H ₂ O) in 40% distilled water was dissolved in MeOH and stirred at room temperature for 30 minutes. At 0 ° C., 0.5 equivalents of sodium borohydride (NaBH ₄ ) were slowly added and stirred for 1 hour. Then, H ₂ O was added to the mixture, MeOH was distilled off under reduced pressure, and extracted three times with dichloromethane (DCM). The DCM layer was dried over NaSO ₄ , filtered, and the solvent was removed by distillation under reduced pressure to obtain 6a-h in a yield of 32.0-76.4%.

2. Amine Synthesis Method II ( 8a-c )

1 equivalent of each benzoic acid, 1.5 equivalents of methylamine in 40% distilled water, 2 equivalents of EDC (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide) or DCC ( N, N -dicyclohexyl-carbodiimide), HOBt (hydroxybenzotriazole 1 equivalent, 1 equivalent of DIPEA ( N, N -diisopropylpropylamine) was dissolved in dimethylformamide (dimethylformamide, DMF), and microwave reaction was performed at 120 ° C. and 20 bar for 3 hours. After dissolving in ethyl acetate (EA) and washing with saturated aqueous 1N HCl solution several times, the EA layer was dried over NaSO ₄ , filtered, and the solvent was removed by distillation under reduced pressure. Then, purified by MPLC or column chromatography, and dissolved 1 equivalent of the purified compound in tetrahydrofuran (THF), and slowly added 3 equivalents of lithium aluminum hydride (LAH) at 0 ° C. Stir for hours. After quenching with 10% aqueous saturated NaOH solution and H ₂ O (quenching), the mixture was extracted using ether. The ether layer was dried over Na ₂ SO ₄ , filtered and the solvent was removed by distillation under reduced pressure. After purification through column chromatography, 8a-c was obtained in a yield of 22.5%, 88.7%.

Compounds 14a to 14i were synthesized in the same manner as in Scheme 2 below.

Scheme 2

3. methyl 2,4- Dihydroxybenzoate [Methyl 2,4- dihydroxybenzoate ] ( 9 )

5 mL of 2,4-Dihydroxybenzoic acid (10.00 g, 64.88 mmol) and sulfuric acid (H ₂ SO ₄ ) were added to 40 mL of MeOH and refluxed at 100 ° C. for 24 hours. ) After cooling to room temperature, the solvent was distilled off under reduced pressure and H ₂ O was added at 0 ° C. The resulting white solid was dissolved in EA and washed with saturated aqueous NaHCO ₃ . The EA layer was dried over Na ₂ SO ₄ , filtered, and the solvent was removed by distillation under reduced pressure to obtain Compound 9 in a yield of 85%.

¹ H NMR (400 MHz, CDCl ₃ ) d 11.0 (s, 1H), 7.73 (d, J = 8.4 Hz, 1H), 6.41-6.37 (m, 2H), 5.78 (s, 1H), 3.92 (s, 3H).

4. methyl 5- Chloro -2,4- Dihydroxybenzoate [Methyl 5- chloro -2,4-dihydroxybenzoate] ( 10 )

Compound 8 (8.30 g, 49.41 mmol) and sulfuryl chloride (SO ₂ Cl ₂ ) (4.11 mL, 56.83 mmol) were added to DCM and stirred at room temperature for 24 hours. The solvent was distilled off under reduced pressure, dissolved in EA, washed with a saturated aqueous NaHCO ₃ solution, the EA layer was dried over Na ₂ SO ₄ , filtered, and the solvent was removed by distillation under reduced pressure. This was purified by column chromatography using a solvent mixed in a ratio of EA: hexane = 1: 9 to obtain a compound 10 in a yield of 32.5%.

¹ H NMR (400 MHz, CDCl ₃ ) d 10.8 (s, 1H), 7.82 (s, 1H), 6.61 (s, 1H), 5.91 (s, 1H), 3.92 (s, 3H).

5. methyl 2,4- Vis ( Allyloxy ) -5- Chlorobenzoate [Methyl 2,4-bis ( allyloxy ) -5-chlorobenzoate] ( 11 )

Compound 9 (1.10 g, 5.46 mmol), allyl bromide (1.23 mL, 14.20 mmol) and K ₂ CO ₃ (1.96 g, 14.20 mmol) were dissolved in DMF. It was stirred for 12 hours at room temperature under argon injection and then dissolved in EA and washed with a saturated aqueous NaHCO ₃ solution. After drying the EA layer with Na ₂ SO ₄ , the solvent was removed by distillation under reduced pressure to obtain Compound 11 in 83% yield.

¹ H NMR (400 MHz, CDCl ₃ ) d 7.85 (s, 1H), 7.25 (s, 1H), 6.44-5.95 (m, 2H), 5.50-5.40 (m, 2H), 5.30-5.26 (m, 2H ), 4.58-4.45 (m, 4H), 3.81 (s, 3H).

6. 2,4- Vis ( Allyloxy ) -5- Chlorobenzoic acid [2,4- Bis ( allyloxy ) -5- chlorobenzoic acid] ( 12 )

Compound 8 (1.27 g, 5.64 mmol) and NaOH (1.12 g, 28.1 mmol) of H ₂ O (20 mL) and 3N HCl saturated aqueous solution into a MeOH (20 mL) is dissolved and then was stirred at room temperature for 8 hours them to EA Washed with. After drying the EA layer with Na ₂ SO ₄ , the solvent was removed by distillation under reduced pressure to obtain Compound 12 in 74% yield.

¹ H NMR (400 MHz, CDCl ₃ ) d 8.16 (s, 1H), 6.54 (s, 1H), 6.10-5.99 (m, 2H), 5.51-5.35 (m, 2H), 4.76-4.65 (m, 4H ).

7. N- benzyl -5- Chloro -2,4- Dihydroxybenzamide [N-Benzyl-5- chloro -2,4-dihydroxybenzamide] ( 14a )

Compound 12 (0.50 g, 1.86 mmol), Benzylamine (0.26 g, 1.86 mmol), HOBt (0.25 g, 1.86 mmol), DCC (0.77 g, 3.72 mmol), DIPEA (0.32 mL, 1.86 mmol) It was dissolved in DMF and microwave reaction at 80 ° C. and 20 bar for 3 hours. The DMF was removed by distillation under reduced pressure, and then purified by MPLC (R _f = 0.19, EA: hexane = 1: 4) to obtain compound 13a . After dissolving compound 13a in THF, PdCl ₂ (PPh ₃ ) ₂ (20 mg) and NH ₄ HCO ₂ (200 mg) were added and microwave-reacted at 120 ° C. and 20 bar for 30 minutes.

After dissolving in EA and washing with H ₂ O, the EA layer was dried over Na ₂ SO ₄ , filtered, the solvent was removed by distillation under reduced pressure, and purified by MPLC (R _f = 0.13, EA: hexane = 3: 7) to 53.2 Compound 14a was obtained in a yield of%.

¹ H NMR (400 MHz, MeOD) d 7.77 (s, 1 H), 7.29-7.26 (m, 4 H), 7.22-7.19 (m, 1 H), 6.40 (s, 1 H), 4.50 (s, 2H). ¹³ C NMR (100 MHz, MeOD) d 165.2, 157.0, 154.2, 135.3, 125.2, 124.7, 123.7, 123.4, 108.1, 104.9, 100.2, 39.1.

8. N- benzyl -5- Chloro -2,4-dihydroxy-N- Methylbenzamide [N-Benzyl-5- chloro -2,4-dihydroxy-N-methylbenzamide] ( 14b )

Compound 12 (0.30 g, 1.12 mmol), Compound 6a (0.21 g, 1.67 mmol), HOBt (0.15 g, 1.12 mmol), DCC (0.46 g, 2.23 mmol), DIPEA (0.19 mL, 1.12 mmol) was dissolved in DMF. The reaction was microwave at 80 ° C., 20 bar for 3 hours. Then, dissolved in EA and washed with H ₂ O, the EA layer was dried over NaSO ₄ , filtered, and the solvent was removed by distillation under reduced pressure. This was purified by MPLC (R _f = 0.31, EA: hexane = 3: 7) to obtain compound 13b . After dissolving compound 13b in THF, PdCl ₂ (PPh ₃ ) ₂ (20 mg) and NH ₄ HCO ₂ (200 mg) were added and microwave-reacted at 120 ° C. and 20 bar for 30 minutes. After dissolving in EA and washing with H ₂ O, the EA layer was dried over Na ₂ SO ₄ , filtered and the solvent was removed by distillation under reduced pressure. This was purified by MPLC (R _f = 0.28, EA: hexane = 2: 3) to obtain Compound 14b in a yield of 62.5%.

¹ H NMR (400 MHz, MeOD) d 7.35-7.24 (m, 5H), 7.16 (s, 1H), 6.50 (s, 1H), 2.89 (s, 3H).

9. 5- Chloro -2,4-dihydroxy-N- (4-methoxybenzyl) -N- Methylbenzamide [5- chloro -2,4-dihydroxy-N- (4-methoxybenzyl) -N-methylbenzamide] ( 14c )

Compound 12 (0.30 g, 1.12 mmol), Compound 6b (0.18 g, 1.67 mmol), HOBt (0.15 g, 1.12 mmol), DCC (0.46 g, 2.23 mmol), DIPEA (0.19 mL, 1.12 mmol) was dissolved in DMF. The microwave reaction was performed at 80 ° C. and 20 bar for 3 hours. The DMF was removed by distillation under reduced pressure, and then purified by MPLC (R _f = 0.20, EA: hexane = 3: 7) to obtain compound 13c . Compound 13c was dissolved in THF, and then PdCl ₂ (PPh ₃ ) ₂ (20 mg) and NH ₄ HCO ₂ (200 mg) were added and subjected to microwave reaction at 120 ° C. and 20 bar for 30 minutes. After dissolving in EA and washing with H ₂ O, the EA layer was dried over Na ₂ SO ₄ , filtered and the solvent was removed by distillation under reduced pressure.

This was purified by MPLC (Rf = 0.30, EA: hexane = 2: 3) to obtain compound 14c in a yield of 63.8%.

¹ H NMR (400 MHz, CDCl ₃ ) d 7.27 (s, 1H), 7.20 (d, J = 8.4 Hz, 2H), 6.90 (d, J = 6.8 Hz, 2H), 6.62 (s, 1H), 4.64 (s, 2H), 3.80 (s, 3H), 3.03 (s, 3H). ¹³ C NMR (100 MHz, CDCl ₃ ) d 171.1, 159.6, 159.3, 155.2, 129.0, 128.8, 128.0, 114.4, 111.1, 110.5, 105.2, 55.4.

10. 5- Chloro -N- (3,4- Dimethoxybenzyl ) -2,4-dihydroxy-N- Methylbenzamide [5-chloro-N- (3,4-dimethoxybenzyl) -2,4-dihydroxy-N-methylbenzamide] ( 14d )

Compound 12 (0.33 g, 1.23 mmol), Compound 6c (0.33 g, 1.84 mmol), HOBt (0.17 g, 1.23 mmol), DCC (0.51 g, 2.46 mmol), DIPEA (0.22 mL, 1.23 mmol) was dissolved in DMF. The microwave reaction was carried out at 120 ° C. and 20 bar for 3 hours. After dissolving in EA and washing with H ₂ O, the EA layer was dried over Na ₂ SO ₄ and filtered, and the solvent was removed by distillation under reduced pressure and purified by MPLC (R _f = 0.26, EA: hexane = 2: 3) to obtain a compound. 13d was obtained. Compound 13d was dissolved in THF, and then PdCl ₂ (PPh ₃ ) ₂ (20 mg) and NH ₄ HCO ₂ (200 mg) were added and subjected to microwave reaction at 120 ° C. and 20 bar for 30 minutes. After dissolving in EA and washing with H ₂ O, the EA layer was dried over Na ₂ SO ₄ , filtered, the solvent was removed by distillation under reduced pressure, and purified by MPLC (R _f = 0.22, EA: hexane = 1: 1) to 63.8%. Compound 14d was obtained in the yield of.

¹ H NMR (400 MHz, CDCl ₃ ) d 7.31 (s, 1H), 6.88-6.81 (m, 3H), 6.66 (s, 1H), 4.65 (s, 2H), 3.88 (d, J = 6.4 Hz, 6H), 3.07 (s, 3 H). ¹³ C NMR (100 MHz, CDCl ₃ ) d 170.5, 155.3, 149.1, 148.3, 131.9, 131.8, 128.8, 128.7, 128.6, 128.5, 119.3, 111.0, 110.5, 105.1, 55.7.

11.N- (benzo [d] [[1,3] dioxol-5-ylmethyl) -5-chloro-2,4-dihydroxy-N-methylbenzamide [N- (benzo [d] [ 1,3] dioxol-5-ylmethyl) -5-chloro-2,4-dihydroxy-N-methylbenzamide] ( 14e )

Compound 12 (0.30 g, 1.12 mmol), Compound 6d (0.28 g, 1.67 mmol), HOBt (0.15 g, 1.12 mmol), DCC (0.46 g, 2.23 mmol), DIPEA (0.19 mL, 1.12 mmol) was dissolved in DMF. The microwave reaction was performed at 80 ° C. and 20 bar for 3 hours. The DMF was removed by distillation under reduced pressure, and then purified by MPLC (R _f = 0.25, EA: hexane = 3: 7) to obtain compound 13e . Compound 13e was dissolved in THF, and then PdCl ₂ (PPh ₃ ) ₂ (20 mg) and NH ₄ HCO ₂ (200 mg) were added and microwave reaction was performed at 120 ° C. and 20 bar for 30 minutes. After dissolving in EA and washing with H ₂ O, the EA layer was dried over Na ₂ SO ₄ , filtered, the solvent was removed by distillation under reduced pressure, and purified by MPLC (R _f = 0.23, EA: hexane = 2: 3) 78.6% Compound 14e was obtained in the yield of.

¹ H NMR (400 MHz, CDCl ₃ ) d 7.28 (s, 1H), 6.79-6.72 (m, 3H), 6.60 (s, 1H), 5.96 (s, 2H), 4.59 (s, 2H), 3.03 ( s, 3H). ¹³ C NMR (100 MHz, CDCl ₃ ) d

12. 5- Chloro -2,4-dihydroxy-N- methyl -N- (3,4,5- Trimethoxybenzyl Benzamide [5-chloro-2,4-dihydroxy-N-methyl-N- (3,4,5-trimethoxybenzyl) benzamide] ( 14f )

Compound 12 (0.30 g, 1.12 mmol), Compound 6e (0.35 g, 1.23 mmol), HOBt (0.15 g, 1.12 mmol), DCC (0.46 g, 2.23 mmol), DIPEA (0.19 mL, 1.12 mmol) was dissolved in DMF. The reaction was microwave at 80 ° C., 20 bar for 3 hours. After dissolving in EA and washing with H ₂ O, the EA layer was dried over NaSO ₄ , filtered, the solvent was removed by distillation under reduced pressure, and purified by MPLC (R _f = 0.20, EA: hexane = 2: 3) to obtain Compound 13f . Got it. Compound 13f was dissolved in THF, and then PdCl ₂ (PPh ₃ ) ₂ (20 mg) and NH ₄ HCO ₂ (200 mg) were added and microwave reaction was performed at 120 ° C. and 20 bar for 30 minutes. After dissolving in EA and washing with H ₂ O, the EA layer was dried over Na ₂ SO ₄ , filtered, the solvent was removed by distillation under reduced pressure, purified by MPLC (R _f = 0.26, EA: hexane = 3: 2), and 67.0. Compound 14f was obtained in the yield of%.

¹ H NMR (400 MHz, CDCl ₃ ) d 7.23 (s, 1H), 6.56 (s, 1H), 6.47 (s, 1H), 4.59 (s, 2H), 3.80 (d, J = 1.2 Hz, 9H) , 3.02 (s, 3 H). ¹³ C NMR (100 MHz, CDCl ₃ ) d 171.2, 158.7, 155.4, 153.6, 137.3, 132.0, 128.8, 111.6, 110.8, 105.1, 104.5, 61.0, 56.2.

13. 5-chloro-N- (2-chloro-3,4,5-trimethoxybenzyl) -2,4-dihydroxy-N-methylbenzamide [5-chloro-N- (2-chloro- 3,4,5-trimethoxybenzyl) -2,4-dihydroxy-N-methylbenzamide] ( 14 g )

Compound 12 (0.20 g, 0.74 mmol), Compound 6f (0.27 g, 1.12 mmol), HOBt (0.10 g, 0.74 mmol), DCC (0.31 g, 1.49 mmol), DIPEA (0.15 mL, 0.74 mmol) was dissolved in DMF. The microwave reaction was carried out at 120 ° C. and 20 bar for 3 hours. After dissolving in EA and washing with H ₂ O, the EA layer was dried over NaSO ₄ , filtered, the solvent was removed by distillation under reduced pressure, and purified by MPLC (R _f = 0.26, EA: hexane = 3: 7) to obtain 13 g of a compound. . 13 g of compound was dissolved in THF, PdCl ₂ (PPh ₃ ) ₂ (20 mg) and NH ₄ HCO ₂ (200 mg) were added, microwave reaction was performed at 120 ° C. and 20 bar for 30 minutes, and dissolved in EA. H ₂ O After washing with, the EA layer was dried over Na ₂ SO ₄ , filtered and the solvent was removed by distillation under reduced pressure. This was purified by MPLC (R _f = 0.20, EA: hexane = 1: 1) to obtain a compound 14g with a yield of 41.9%.

¹ H NMR (400 MHz, CDCl ₃ ) d 7.21 (s, 1H), 6.75 (s, 1H), 6.47 (s, 1H), 4.75 (s 2H), 3.90 (d, J = 9.2 Hz, 6H), 3.83 (s, 3 H), 3.03 (s, 3 H). ¹³ C NMR (100 MHz, CDCl ₃ ) d 171.1, 156.3, 155.4, 152.5, 150.0, 142.4, 132.0, 129.3, 128.9, 119.2, 113.0, 111.3, 107.0, 104.2, 61.2, 56.1, 49.8.

14.N- (2-bromo-3,4,5-trimethoxybenzyl) -5-chloro-2,4-dihydroxy-N-methylbenzamide [N- (2-bromo-3,4 , 5-trimethoxybenzyl) -5-chloro-2,4-dihydroxy-N-methylbenzamide] ( 14h )

Compound 12 (0.19 g, 0.69 mmol), Compound 6g (0.30 g, 1.03 mmol), HOBt (0.09 g, 0.69 mmol), DCC (0.28 g, 1.38 mmol), DIPEA (0.12 mL, 0.69 mmol) was dissolved in DMF. The microwave reaction was carried out at 120 ° C. and 20 bar for 3 hours. After dissolving in EA and washing with H ₂ O, the EA layer was dried over Na ₂ SO ₄ , filtered, the solvent was removed by distillation under reduced pressure, and purified by MPLC (R _f = 0.24, EA: hexane = 3: 7) to obtain a compound. 13h was obtained.

Compound 13h was dissolved in THF, and then PdCl ₂ (PPh ₃ ) ₂ (20 mg) and NH ₄ HCO ₂ (200 mg) were added and microwave reaction was performed at 120 ° C. and 20 bar for 30 minutes. After dissolving in EA and washing with H ₂ O, the EA layer was dried over Na ₂ SO ₄ , filtered, the solvent was removed by distillation under reduced pressure, and purified by MPLC (R _f = 0.24, EA: hexane = 1: 1) to 20.5. Compound 14h was obtained with a yield of%.

¹ H NMR (400 MHz, CDCl ₃ ) d 7.15 (s, 1H), 6.69 (s, 1H), 6.42 (s, 1H), 4.68 (s, 2H), 3.83 (d, J = 6.0 Hz, 6H) , 3.77 (d, 3 H), 2.98 (s, 3 H). ¹³ C NMR (100 MHz, CDCl ₃ ) d 160.8, 155.2, 153.4, 151.4, 143.0, 130.9, 128.6, 110.7, 110.3, 105.4, 61.4, 61.3, 60.7, 56.5.

15. 5- Chloro -2,4-dihydroxy-N- methyl -N- (3- Methylbenzyl Benzamide [5- chloro -2,4-dihydroxy-N-methyl-N- (3-methylbenzyl) benzamide] ( 14i )

Compound 12 (0.25 g, 0.93 mmol), Compound 6h (0.19 g, 1.40 mmol), HOBt (0.13 g, 0.93 mmol), DCC (0.39 g, 1.86 mmol), DIPEA (0.17 mL, 0.93 mmol) was dissolved in DMF. The microwave reaction was carried out at 120 ° C. and 20 bar for 3 hours. After dissolving in EA and washing with H ₂ O, the EA layer was dried over NaSO ₄ and filtered, the solvent was removed by distillation under reduced pressure, and purified by MPLC (R _f = 0.22, EA: hexane = 1: 4) to compound 13i. Got.

Compound 13i was dissolved in THF, PdCl ₂ (PPh ₃ ) ₂ (20 mg) and NH ₄ HCO ₂ (200 mg) were added and microwave reaction was performed at 120 ° C. and 20 bar for 30 minutes. After dissolving in EA and washing with H ₂ O, the EA layer was dried over Na ₂ SO ₄ and filtered, the solvent was removed by distillation under reduced pressure, and purified by MPLC (R _f = 0.20, EA: hexane = 3: 7). Compound 14i was obtained with a yield of 68.7%.

¹ H NMR (400 MHz, CDCl ₃ ) d 7.31 (s, 1H), 7.28 (d, J = 7.6 Hz, 1H), 7.15 (d, J = 7.6 Hz, 1H), 7.10-7.09 (m, 2H) , 6.65 (s, 1H), 4.70 (s, 2H), 3.08 (s, 3H), 2.38 (s, 3H). ¹³ C NMR (100 MHz, CDCl ₃ ) d 171.3, 160.0, 155.2, 138.8, 136.0, 128.9, 128.8, 128.7, 128.3, 124.6, 110.9, 110.4, 105.2, 36.8.

Compounds 16a and 16b were synthesized in the same manner as in Scheme 3 below.

Scheme 3

16. 5- Chloro -2,4- Dihydroxybenzoic acid [5- chloro -2,4- dihydroxybenzoic acid] ( 15 )

Compound 9 (1.98 g, 9.77 mmol) and NaOH (1.95 g, 48.80 mmol) were added to H ₂ O (30 mL) and MeOH (30 mL) and stirred at room temperature for 12 hours. This was dissolved in EA, washed with a saturated 3N HCl aqueous solution, the EA layer was dried over Na ₂ SO ₄ , and the solvent was removed by distillation under reduced pressure to obtain Compound 15 in a yield of 100%.

¹ H NMR (400 MHz, CDCl ₃ ) d 7.76 (s, 1H), 6.41 (s, 1H).

17. 5- Chloro -2,4-dihydroxy-N- methyl -N- (2- Methylbenzyl Benzamide [5- chloro -2,4-dihydroxy-N-methyl-N- (2-methylbenzyl) benzamide] ( 16a )

Compound 15 (0.17 g, 0.92 mmol), Compound 8a (0.19 g, 1.40 mmol), HOBt (0.12 g, 0.92 mmol), EDC (0.35 g, 1.86 mmol), DIPEA (0.16 mL, 0.92 mmol) was dissolved in DMF. The microwave reaction was carried out at 120 ° C. and 20 bar for 3 hours. This was dissolved in EA, washed with 1N HCl saturated aqueous solution, the EA layer was dried over Na ₂ SO ₄ , filtered, and the solvent was removed by distillation under reduced pressure. Then purified by MPLC (R _f = 0.23, EA: hexane = 1: 4) to give the compound 16a in 90.5% yield.

¹ H NMR (400 MHz, CDCl ₃ ) d 7.25-7.24 (m, 4H), 6.65 (s, 1H), 4.72 (s, 2H), 3.07 (s, 3H), 2.27 (s, 3H). ¹³ C NMR (100 MHz, CDCl ₃ ) d 160.9, 155.2, 136.4, 134.0, 131.0, 128.7, 128.0, 127.4, 126.7, 110.7, 110.2, 105.3, 19.2.

18. 5- Chloro -2,4-dihydroxy-N- methyl -N- (4- Methylbenzyl Benzamide [5- chloro -2,4-dihydroxy-N-methyl-N- (4-methylbenzyl) benzamide] ( 16b )

Compound 15 (0.21 g, 1.12 mmol), Compound 8b (0.23 g, 1.69 mmol), HOBt (0.15 g, 1.12 mmol), EDC (0.43 g, 2.25 mmol), DIPEA (0.20 mL, 1.12 mmol) was dissolved in DMF. The microwave reaction was carried out at 120 ° C. and 20 bar for 3 hours. It was dissolved in EA, washed with saturated aqueous 1N HCl solution, the EA layer was dried over Na ₂ SO ₄ , filtered, and the solvent was removed by distillation under reduced pressure. Then purified by MPLC (R _f = 0.23, EA: hexane = 1: 4) to give the compound 16b in a yield of 26.0%.

¹ H NMR (400 MHz, CDCl ₃ ) d 7.28 (s, 1H), 7.17 (t, J = 9.2 Hz, 9.2 Hz, 4H), 6.63 (s, 1H), 4.67 (s, 2H), 3.04 (s , 3H), 2.35 (s, 3H). ¹³ C NMR (100 MHz, CDCl ₃ ) d 160.1, 155.0, 137.7, 133.0, 129.7, 128.6, 128.5, 127.6, 127.3, 110.9, 110.2, 105.2, 21.2.

Compounds 21a to 21f were synthesized in the same manner as in Scheme 4 below.

Scheme 4

19. methyl 2,4-dihydroxy-5- Isopropylbenzoate [Methyl 2,4- dihydroxy -5-isopropylbenzoate] ( 17 )

Compound 9 (10.70 g, 63.50 mmol), AlCl ₃ (16.90 g, 127.0 mmol) and 2-bromopropane (11.90 mL, 127.0 mmol) were dissolved in DCM (125 mL) and refluxed at 50 ° C. for 24 hours under argon injection. 2 equivalents of AlCl ₃ and 2-bromopropane were added every 6 hours. Adjust to pH 6 with 10% saturated aqueous NaOH solution, remove the solvent by distillation under reduced pressure, and wash with saturated NaHCO ₃ aqueous solution. It was. The EA layer was dried over Na ₂ SO ₄ , filtered, and the solvent was removed by distillation under reduced pressure. Purification by column chromatography using a solvent mixed in a ratio of EA: hexane = 1: 9 to give a compound 17 in a yield of 34.5%.

¹ H NMR (400 MHz, CDCl ₃ ) d 12.61 (s, 1H), 7.52 (s, 1H), 6.35 (s, 1H), 6.31 (s, 1H), 3.22-3.12 (m, 1H), 2.61 ( s, 3H), 1.27 (d, J = 6.8 Hz, 6H). ¹³ C NMR (100 MHz, CDCl ₃ ) d 170.7, 161.6, 159.6, 128.1, 127.1, 105.7, 103.2, 52.2, 26.7, 22.8.

20. 2,4-dihydroxy-5- Isopropylbenzoic acid [2,4- Dihydroxy -5- isopropylbenzoic acid] ( 18 )

Compound 17 (4.60 g, 21.90 mmol) and LiOH (10 g) were added to H ₂ O (30 mL) and MeOH (30 mL) at 70 ° C. and refluxed for 12 hours. After dissolving in EA and washing with 3N HCl saturated aqueous solution, the EA layer was dried over Na ₂ SO ₄ , filtered, and the solvent was removed by distillation under reduced pressure to obtain Compound 18 in a yield of 60.5%.

¹ H NMR (400 MHz, CDCl ₃ ) d 7.68 (s, 1H), 6.30 (s, 1H), 3.19-3.12 (m, 1H), 1.17 (d, J = 4.0 Hz, 6H).

21. methyl 4-((5-chloro-2,4-dihydroxy-N-methylbenzamido) methyl) benzoate [Methyl 4-((5- chloro -2,4- dihydroxy -N- methylbenzamido ) methyl) benzoate] ( 19a ) And methyl 4-((2,4-dihydroxy-5-isopropyl-N- Methylbenzamido ) methyl ) Benzoa Tri [methyl 4-((2,4- dihydroxy -5-isopropyl-N- methylbenzamido ) methyl) benzoate] ( 19b )

1 equivalent of Compound 12 or 18 , 1.5 equivalent of Compound 8c , 1 equivalent of HOBt, 2 equivalents of EDC and 1 equivalent of DIPEA were dissolved in DMF and microwave reacted at 120 ° C. and 20 bar for 3 hours. After dissolving in EA and washing with H ₂ O, the EA layer was dried over NaSO ₄ , filtered and the solvent was removed by distillation under reduced pressure. Purification by MPLC according to the conditions of each compound to give the compound 19a or 19b in 76.1%, 81.7% yield.

19a : ¹ H NMR (500 MHz, CDCl ₃ ) d 8.03 (d, J = 6.6 Hz, 2H), 7.35 (d, J = 6.6 Hz, 2H), 7.25 (s, 1H), 6.62 (s, 1H) , 4.75 (s, 2H), 3.91 (s, 3H), 3.07 (s, 3H).

19b : ¹ H NMR (500 MHz, CDCl ₃ ) d 8.05 (d, J = 6.7 Hz, 2H), 7.37 (d, J = 6.7 Hz, 2H), 7.11 (s, 1H), 6.42 (s, 1H) , 4.76 (s, 2H), 3.92 (s, 3H), 3.13-3.04 (m, 1H), 3.05 (s, 3H), 0.94 (d, J = 5.3 Hz, 6H).

22. 4-((5- Chloro -2,4-dihydroxy-N- Methylbenzamido ) methyl Benzoic acid [4-((5-chloro-2,4-dihydroxy-N-methylbenzamido) methyl) benzoic acid] ( 20a ) And 4-((2,4-dihydroxy-5-isopropyl-N-methylbenzamido) methyl) benzoic acid (4-((2,4-dihydroxy-5-isopropyl-N-methylbenzamido) methyl) benzoic acid] ( 20b )

1 equivalent of Compound 19a or 19b and LiOH (2.0 g) were added to H ₂ O (20 mL) and MeOH (20 mL) and stirred at room temperature for 3 hours. After dissolving in EA and washing with 3N HCl saturated aqueous solution, the EA layer was dried over Na ₂ SO ₄ and filtered, and the solvent was removed by distillation under reduced pressure to obtain the compound 20a, 20b in 84.8% and 88.1% yield.

20a : ¹ H NMR (500 MHz, CDCl ₃ ) d 8.13 (d, J = 6.66 Hz, 2H), 7.41 (d, J = 6.6 Hz, 2H), 7.30 (s, 1H), 6.68 (s, 1H) , 4.79 (s, 2 H), 3.12 (s, 3 H).

20b : ¹ H NMR (500 MHz, MeOD) d 8.00 (d, J = 6.5 Hz, 2H), 7.41 (d, J = 5.8 Hz, 2H), 7.0 (s, 1H), 6.34 (s, 1H), 4.86 (s, 2H), 3.18-3.11 (m, 1H), 3.0 (s, 3H), 1.11 (d, J = 5.2 Hz, 6H).

23. 5- Chloro -2,4-dihydroxy-N- methyl -N- (4- ( Methylcarbamoyl ) benzyl Benzamide [5-chloro-2,4-dihydroxy-N-methyl-N- (4- (methylcarbamoyl) benzyl) benzamide] ( 21a )

Compound 20a (0.27 g, 0.79 mmol), methylamine (0.10 mL, 1.18 mmol) in 40% distilled water, HOBt (0.11 g, 0.79 mmol), EDC (0.30 g, 1.58 mmol), DIPEA (0.14 mL, 0.79 mmol) It was dissolved in DMF and microwave reaction at 120 ℃, 20 bar for 2 hours. This was dissolved in EA, washed with 1N HCl saturated aqueous solution, the EA layer was dried over Na ₂ SO ₄ , filtered, and the solvent was removed by distillation under reduced pressure. Then purified by MPLC (C18 column, R _f = 0.21, H ₂ O: MeOH = 4: 1) to give the compound 21a in a yield of 16.7%.

¹ H NMR (500 MHz, CDCl ₃ ) d 7.78 (d, J = 6.6 Hz, 2H), 7.36 (d, J = 6.6 Hz, 2H), 7.30 (s, 1H), 6.68 (s, 1H), 4.75 (s, 2H), 3.09 (s, 3H), 3.03 (d, J = 3.8 Hz, 3H).

24. 5- Chloro -N- (4- ( Ethylcarbamoyl ) benzyl ) -2,4-dihydroxy-N- Methylbenzami De [5-chloro-N- (4- (ethylcarbamoyl) benzyl) -2,4-dihydroxy-N-methylbenzamide] ( 21b )

Compound 20a (0.27 g, 0.79 mmol), Ethylamine in H ₂ O (0.10 mL, 1.18 mmol), HOBt (0.11 g, 0.79 mmol), EDC (0.30 g, 1.58 mmol), DIPEA (0.14 mL, 0.79 mmol) was dissolved in DMF and microwave reacted at 120 ° C. and 20 bar for 2 hours. After dissolving in EA and washing with saturated aqueous 1N HCl solution, the EA layer was dried over Na ₂ SO ₄ , filtered, and the solvent was removed by distillation under reduced pressure. Then purified by MPLC (C18 column, _Rf = 0.21, H ₂ O: MeOH = 4: 1) to give the compound 21b in a yield of 31.2%.

¹ H NMR (500 MHz, CDCl ₃ ) d 7.78 (d, J = 6.6 Hz, 2H), 7.35 (d, J = 6.6 Hz, 2H), 7.30 (s, 1H), 6.67 (s, 1H), 4.75 (s, 2H), 3.54-3.49 (m, 2H), 2.96 (s, 3H), 1.26 (t, J = 5.8 Hz, 5.8 Hz, 3H). ¹³ C NMR (100 MHz, CDCl ₃ ) d

25. 5- Chloro -2,4-dihydroxy-N- methyl -N- (4- ( Profile Caba Mole ) benzyl Benzamide [5-chloro-2,4-dihydroxy-N-methyl-N- (4- (propylcarbamoyl) benzyl) benzamide] ( 21c )

Compound 20a (0.26 g, 0.76 mmol), Propylamine (0.10 mL, 1.14 mmol), HOBt (0.10 g, 0.76 mmol), EDC (0.30 g, 1.52 mmol), DIPEA (0.10 mL, 0.76 mmol) It was dissolved in DMF and microwave reaction at 120 ° C. and 20 bar for 3 hours. It was dissolved in EA, washed with 1N HCl saturated aqueous solution, the EA layer was dried over NaSO ₄ , filtered, and the solvent was removed by distillation under reduced pressure. Then purified by MPLC (C18 column, R _f = 0.21, H ₂ O: MeOH = 7: 3) to give the compound 21c in a yield of 45.5%.

¹ H NMR (500 MHz, CDCl ₃ ) d 7.78 (d, J = 6.6 Hz, 2H), 7.35 (d, J = 6.6 Hz, 2H), 7.28 (s, 1H), 6.67 (s, 1H), 4.75 (s, 2H), 3.46-3.42 (m, 2H), 3.09 (s, 3H), 1.67-1.62 (m, 2H), 1.00 (t, J = 5.9 Hz, 5.9 Hz, H).

26. 2,4-dihydroxy-5-isopropyl-N- methyl -N- (4- ( Methylcarbamoyl ) benzyl ) Benzamide [2,4-dihydroxy-5-isopropyl-N-methyl-N- (4- (methylcarbamoyl) benzyl) benzamide] ( 21d )

Compound 20b (0.21 g, 0.62 mmol), methylamine (0.08 mL, 0.93 mmol) in 40% distilled water, HOBt (0.08 g, 0.62 mmol), EDC (0.23 g, 1.24 mmol), DIPEA (0.11 mL, 0.62 mmol) It was dissolved in DMF and microwave reaction at 120 ℃, 20 bar for 3 hours. This was dissolved in EA, washed with 1N HCl saturated aqueous solution, the EA layer was dried over Na ₂ SO ₄ , filtered, and the solvent was removed by distillation under reduced pressure. Then purified by MPLC (C18 column, R _f = 0.21, H ₂ O: MeOH = 7: 3) to give the compound 21d in a yield of 36.1%.

¹ H NMR (500 MHz, CDCl ₃ ) d 7.79 (d, J = 6.5 Hz, 2H), 7.36 (d, J = 6.4 Hz, 2H), 7.08 (s, 1H), 6.41 (s, 1H), 6.46 (s, 2H), 3.08 (s, 3H), 3.06-3.03 (m, 1H), 0.98 (d, J = 5.3 Hz, 6H).

27.N- (4- ( Ethylcarbamoyl N- (4- (ethylcarbamoyl) benzyl) -2,4-dihydroxy-5-isopropyl-N-methylbenzamide] (benzyl) -2,4-dihydroxy-5-isopropyl-N-methylbenzamide 21e )

Compound 20b (0.22 g, 0.64 mmol), ethylamine (0.08 mL, 0.97 mmol) in 70% distilled water, HOBt (0.09 g, 0.64 mmol), EDC (0.25 g, 1.29 mmol), DIPEA (0.12 mL, 0.64 mmol) It was dissolved in DMF and microwave reaction at 120 ℃, 20 bar for 3 hours. This was dissolved in EA, washed with 1N HCl saturated aqueous solution, the EA layer was dried over Na ₂ SO ₄ , filtered, and the solvent was removed by distillation under reduced pressure. Then purified by MPLC (C18 column, R _f = 0.21, H ₂ O: MeOH = 7: 3) to give the compound 21e in a yield of 24.3%.

¹ H NMR (500 MHz, CDCl ₃ ) d 7.78 (d, J = 6.6 Hz, 2H), 7.33 (d, J = 6.6 Hz, 2H), 7.05 (s, 1H), 6.43 (s, 1H), 4.73 (s, 2H), 3.52-3.47 (m, 2H), 3.07-3.05 (m, 1H), 3.05 (s, 3H), 1.27-1.23 (m, 3H), 0.98 (d, J = 5.4 Hz, 6H ).

28. 2,4-Dihydroxy-5-isopropyl-N-methyl-N- (4- (propylcarbamoyl) benzyl) benzamide [2,4-dihydroxy-5-isopropyl-N-methyl-N- (4- (propylcarbamoyl) benzyl) benzamide] ( 21f )

Compound 20b (0.22 g, 0.64 mmol), propylamine (0.08 mL, 0.96 mmol), HOBt (0.09 g, 0.64 mmol), EDC (0.25 g, 1.28 mmol), DIPEA (0.11 mL, 0.64 mmol) was dissolved in DMF. The microwave reaction was carried out at 120 ° C. and 20 bar for 3 hours. After dissolving in EA and washing with saturated aqueous 1N HCl solution, the EA layer was dried over Na ₂ SO ₄ , filtered and the solvent was removed by distillation under reduced pressure. Then purified by MPLC (C18 column, R _f = 0.21, H ₂ O: MeOH = 7: 3) to give the compound 21f in a yield of 22. 3%.

¹ H NMR (500 MHz, CDCl ₃ ) d 7.76 (d, J = 6.6 Hz, 2H), 7.29 (d, J = 6.6 Hz, 2H), 7.02 (s, 1H), 6.43 (s, 1H), 4.70 (s, 2H), 3.41-3.37 (m, 2H), 3.07-3.05 (m, 1H), 3.02 (s, 3H), 1.63 (s, 2H), 0.97-0.92 (m, 9H).

29.N- benzyl -2,4-dihydroxy-5-isopropyl-N- Methylbenzamide [N-benzyl-2,4-dihydroxy-5-isopropyl-N-methylbenzamide] ( 21 g )

2,4-dihydroxy-5-isopropylbenzoic acid [2,4-Dihydroxy-5-isopropylbenzoid acid (0.33 g, 1.70 mmol)], N-methylaniline (0.33 g, 2.54 mmol), EDC (0.65 g, 3.30 mmol) and DIPEA (0.30 mL, 1.70 mmol) were dissolved in DMF and microwave reacted at 120 ° C. and 20 bar for 3 hours. After dissolving in EA and washing with saturated aqueous 1N HCl solution, the EA layer was dried over Na ₂ SO ₄ , filtered and the solvent was removed by distillation under reduced pressure. EA: Hexane = 1: 4 (R _f = 0.18) Purified using a mixed solvent, to obtain 21 g of a compound in a yield of 15.5%.

¹ H NMR (500 MHz, CDCl ₃ ) δ10.5 (s, 1H), 7.42-7.39 (m, 2H), 7.34-7.29 (m, 3H), 6.38 (s, 1H), 5.34 (s, 1H) , 4.74 (s, 2H), 3.09 (s, 3H), 3.06-2.95 (m, 1H), 0.96 (d, J = 5.4 Hz, 6H).

< Experimental Example 1> Assessment of Biological Activity of Benzamide Compounds

One. On Hsp90 Bond strength

A fluorescence polarization assay was performed to confirm the binding force according to the concentration of the compound synthesized with Hsp90. 1M dithiothreitol, in hexafluorobenzene (HFB) buffer (20 mM HEPES pH 7.3, 50 mM KCl, 5 mM MgCl ₂ , 20 mM Na ₂ MoO ₄ , 0.01% NP4O, tertiary distilled water) DTT) and 10 mg / mL BGG (bovine gamma globulin) were added and mixed. 100 nM gambogic acid (GA) -FITC (Fluorescein isothiocyanate) was added thereto and reacted at room temperature for 10 minutes. The mixture was dispensed by 100 μL into a control well of a 96 well plate and 2 μL of Hsp90α was added to the remaining mixture, and 100 μL into another control well. 98 μL was dispensed into the wells to be treated with the compound, followed by 2 μL at 5 mM, 2.5 mM, 500 μM, 50 μM, 25 μM, 5 μM, 500 nM or 50 nM for each concentration. After reacting for 1, 4, 6 or 18 hours, the fluorescence polarization was measured at 495/530 nm with a plate leader, and the binding force of the compound to Hsp90 was expressed in mP (millipolarization unit = 1000 mP unit). The results are shown in Table 1 below.

2. Cell viability measurement

MTS [3- (4,5-dimethylthiazol-2-y) -5- (3 to determine the cell viability according to the concentration of the synthesized compound in H1975 cells (ATCC) resistant to Gefitinib (Gefitinib) -carboxymethoxyphenyl) -2- (4-sulfophenyl) -2H-tetrazolium, inner salt] analysis was performed. When the cells were cultured to occupy about 80% of the bottom of the plate, the cells were detached and diluted with RPMI 1640 medium to 1.5 × 10 ³ cells / well. The suspension was placed in 100 μL of each well of a 96 well plate and incubated for 14 hours. The compound is then added at each concentration ( 1a-g : 0, 1, 5, 10, 30, 50 or 100 μM, 14a-i, 16a, b , 21a-f : 0, 0.01, 0.1, 1, 5, 10 , 30, 50, 70 or 100 μM) was changed to the culture solution and then cultured for 3 days. 20 μL of MTS solution was dispensed into each well, followed by reaction for 1 hour in a 37 ° C. incubator supplied with 5% CO ₂ . Absorbance was measured at 490/690 nm for 1 hour with a microplate reader to calculate cell viability according to concentration (%) and IC ₅₀ (μM), and the results are shown in Table 1 below.

[Formula 1]

compound R _One R ₂ Hsp90 ( FP )
( IC ₅₀ ; nM ) H1975
( EC ₅₀ ; μM ) 14a Cl

6683 86.2 14b Cl

413 35.5 14c Cl

135 16.7 14d Cl

362 33.4 14e Cl

585 103 14f Cl

1461 73.1 14 g Cl

429 33.8 14h Cl

528 34.3 14i Cl

995 47.5 16a Cl

830 49.3 16b Cl

558 34.3 21a Cl

496.0 34.8 21b Cl

611.5 21.7 21c Cl

454.0 20.8 21d Isopropyl
(isopropyl)

11.4 0.83 21e Isopropyl

6.5 0.46 21f Isopropyl

5.3 0.42 21 g Isopropyl

33 4.1

As a result of measuring the binding force between the benzamide compounds 14a to 14i, 16a, 16b, and 21a to 21f and Hsp90 according to the present invention, as shown in Table 1 , the compounds showed high binding strength at the nanomolar concentration level except for 14a and 14f . It was. Among them, compound 21f was found to exhibit the best binding force of IC ₅₀ 5.3 nM.

In addition, as a result of measuring the cell viability of the compound in H1975 cells, as shown in Table 1, it was confirmed that the cell proliferation is inhibited by all the benzamide compound in a time-dependent manner, especially compound 21f EC ₅₀ 420 nM good efficacy It was confirmed that it represents.

3. Protein Expression Analysis

We selected a potent compound 21f and confirmed the proliferation effect of Hsp90 client protein in the same cell line.Hsp90 client protein was used as human EGFR-related 2 (Her2), epidermal growth factor receptor (EGFR), Met, and c-Raf. It was.

Incubate H1975 cells resistant to zephytinib for 24 hours in culture medium containing 10% fetal bovin serum (FBS) and detach them, and then RPMI to 5 × 10 ⁵ cells / well. Diluted with 1640 medium. The suspensions were placed in 6 mL petri dishes and incubated for 24 hours. Thus, each compound was changed to a culture solution treated with each concentration ( 21f : 0, 0.05, 0.1, 0.5 or 1 μM, GA : 1 μM), and then cultured for 24 hours. After incubation, the cells were harvested, centrifuged at 4 ° C. and 1300 rpm for 5 minutes, and the supernatant was removed. The pellets were washed with phosphate buffered saline, centrifuged once more under the same conditions, and the supernatant was removed. Lysis buffer was added to the remaining pellets and dissolved by vortexing at 5 minute intervals for 30 minutes. After centrifugation at 4 ℃, 16000 rcf for 40 minutes to separate only the supernatant, the protein was quantified using a BCA (bovine serum albumin) kit. Sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed using 10, 12% running gel and 4% stacking gel (20 mA per gel). The separated protein was transferred to an immunooblot polyvinylidene fluoride membrane at 100 V for 75 minutes. After blocking the membrane with a 5% skim milk solution for 2 hours, the primary antibody diluted at a ratio of 1: 750 to 1,000 was added to a 0.1% tween-20 solution at 4 ° C. Reaction was carried out for 12 hours. This was washed three times with TBS-T (Tris-buffered Saline and Tween 20), and then detected by an image analyzer using an enhanced chemiluminescence (ECL) solution.

As a result, as shown in FIG. 1, it was confirmed that the proliferation of Hsp90 client protein was inhibited by the concentration of compound 21f . In Her2, Met, and Akt, 0.5 μM and EGFR were significantly decreased at 0.1 μM. In addition, Hsp70 also increased expression at 0.5 μM.

As mentioned above, although this invention was demonstrated by the limited embodiment and drawing, this invention is not limited by this, The person of ordinary skill in the art to which this invention belongs, Of course, various modifications and variations are possible within the scope of equivalents of the claims to be described.

Claims

Benzamide compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof:
[Formula 1]

In Chemical Formula 1,
R ₁ is C1-C4 alkyl,
R ₂ is

ego,
R ₃ is any one selected from the group consisting of H, C 1 -C 4 alkyl and C 1 -C 4 alkoxy,
R ₉ is C1-C4 alkyl.

delete

The method of claim 1,
Benzamide compound represented by Chemical Formula 1 is 2,4-dihydroxy-5-isopropyl-N-methyl-N- (4- (methylcarbamoyl) benzyl) benzamide [2,4-dihydroxy-5- isopropyl-N-methyl-N- (4- (methylcarbamoyl) benzyl) benzamide], N- (4- (ethylcarbamoyl) benzyl) -2,4-dihydroxy-5-isopropyl-N-methylbenzamide [N- (4- (ethylcarbamoyl) benzyl) -2,4-dihydroxy-5-isopropyl-N-methylbenzamide], and 2,4-dihydroxy-5-isopropyl-N-methyl-N- (4- (Propylcarbamoyl) benzyl) benzamide [2,4-dihydroxy-5-isopropyl-N-methyl-N- (4- (propylcarbamoyl) benzyl) benzamide] benz, characterized in that any one selected from the group consisting of Amide compounds or pharmaceutically acceptable salts thereof.

A pharmaceutical composition for preventing or treating cancer diseases comprising a benzamide compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient:
[Formula 1]

In Chemical Formula 1,
R ₁ is C1-C4 alkyl,
R ₂ is

delete

The method of claim 4, wherein
Benzamide compound represented by Chemical Formula 1 is 2,4-dihydroxy-5-isopropyl-N-methyl-N- (4- (methylcarbamoyl) benzyl) benzamide [2,4-dihydroxy-5- isopropyl-N-methyl-N- (4- (methylcarbamoyl) benzyl) benzamide], N- (4- (ethylcarbamoyl) benzyl) -2,4-dihydroxy-5-isopropyl-N-methylbenzamide [N- (4- (ethylcarbamoyl) benzyl) -2,4-dihydroxy-5-isopropyl-N-methylbenzamide], and 2,4-dihydroxy-5-isopropyl-N-methyl-N- (4- (Propylcarbamoyl) benzyl) benzamide [2,4-dihydroxy-5-isopropyl-N-methyl-N- (4- (propylcarbamoyl) benzyl) benzamide] A cancer characterized in that it is any one selected from the group consisting of Pharmaceutical composition for preventing or treating disease.

delete

The method of claim 4, wherein
The cancer diseases include non-small cell lung cancer, breast cancer, ovarian cancer, uterine cancer, pancreatic cancer, lung cancer, stomach cancer, liver cancer, colon cancer, skin cancer, head or neck cancer, brain cancer, laryngeal cancer, prostate cancer, bladder cancer, esophageal cancer, thyroid cancer, kidney cancer, and rectal cancer. Acute myeloid leukemia, chronic myelogenous leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia and hematological cancer, characterized in that any one selected from the group consisting of hematological cancer.

delete

A health functional food for preventing or improving cancer diseases comprising a benzamide compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient:
[Formula 1]

In Chemical Formula 1,
R ₁ is C1-C4 alkyl,
R ₂ is

delete

The method of claim 10,
Benzamide compound represented by Chemical Formula 1 is 2,4-dihydroxy-5-isopropyl-N-methyl-N- (4- (methylcarbamoyl) benzyl) benzamide [2,4-dihydroxy-5- isopropyl-N-methyl-N- (4- (methylcarbamoyl) benzyl) benzamide], N- (4- (ethylcarbamoyl) benzyl) -2,4-dihydroxy-5-isopropyl-N-methylbenzamide [N- (4- (ethylcarbamoyl) benzyl) -2,4-dihydroxy-5-isopropyl-N-methylbenzamide], and 2,4-dihydroxy-5-isopropyl-N-methyl-N- (4- (Propylcarbamoyl) benzyl) benzamide [2,4-dihydroxy-5-isopropyl-N-methyl-N- (4- (propylcarbamoyl) benzyl) benzamide] A cancer characterized in that it is any one selected from the group consisting of Health functional food for disease prevention or improvement.

A pharmaceutical composition for preventing or treating degenerative neurological disease, comprising a benzamide compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient:
[Formula 1]

In Chemical Formula 1,
R ₁ is C1-C4 alkyl,
R ₂ is

The method of claim 13,
Benzamide compound represented by Formula 1 is 2,4-dihydroxy-5-isopropyl-N-methyl-N- (4- (methylcarbamoyl) benzyl) benzamide [2,4-dihydroxy-5- isopropyl-N-methyl-N- (4- (methylcarbamoyl) benzyl) benzamide], N- (4- (ethylcarbamoyl) benzyl) -2,4-dihydroxy-5-isopropyl-N-methylbenzamide [N- (4- (ethylcarbamoyl) benzyl) -2,4-dihydroxy-5-isopropyl-N-methylbenzamide], and 2,4-dihydroxy-5-isopropyl-N-methyl-N- (4- (Propyl carbamoyl) benzyl) benzamide [2,4-dihydroxy-5-isopropyl-N-methyl-N- (4- (propylcarbamoyl) benzyl) benzamide], characterized in that it is any one selected from the group consisting of Pharmaceutical composition for preventing or treating neurological diseases.

The method of claim 13, wherein the neurodegenerative disorders include stroke, stroke, memory loss, memory impairment, dementia, forgetfulness, Parkinson's disease, Alzheimer's disease, Pick's disease, Creutzfeld-Kacob disease, Huntington's disease and Pharmaceutical composition for preventing or treating degenerative neurological disease, characterized in that any one selected from the group consisting of Lou Gehrig's disease.

A pharmaceutical composition for preventing or treating a viral infection comprising a benzamide compound represented by the following Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient:
[Formula 1]

In Chemical Formula 1,
R ₁ is C1-C4 alkyl,
R ₂ is

ego,
R ₃ is any one selected from the group consisting of H, C 1 -C 4 alkyl and C 1 -C 4 alkoxy and R ₉ is C 1 -C 4 alkyl.

The method of claim 16,
Benzamide compound represented by Formula 1 is 2,4-dihydroxy-5-isopropyl-N-methyl-N- (4- (methylcarbamoyl) benzyl) benzamide [2,4-dihydroxy-5- isopropyl-N-methyl-N- (4- (methylcarbamoyl) benzyl) benzamide], N- (4- (ethylcarbamoyl) benzyl) -2,4-dihydroxy-5-isopropyl-N-methylbenzamide [N- (4- (ethylcarbamoyl) benzyl) -2,4-dihydroxy-5-isopropyl-N-methylbenzamide], and 2,4-dihydroxy-5-isopropyl-N-methyl-N- (4- (Propyl carbamoyl) benzyl) benzamide [2,4-dihydroxy-5-isopropyl-N-methyl-N- (4- (propylcarbamoyl) benzyl) benzamide], characterized in that the virus is any one selected from the group consisting of Pharmaceutical composition for the prevention or treatment of infectious diseases.