KR20150049695A - Flavokawain derivative or pharmaceutically acceptable salt thereof having inhibitory activity on Hsp90 and medical use thereof - Google Patents

Abstract

The present invention relates to a flavokawain derivative or a pharmaceutically acceptable salt thereof having an activity for inhibiting Hsp90 and a medical use thereof wherein the flavokawain derivative comprises good effect of inhibiting Hsp90 and accordingly leads to decomposition of Hsp 90 client protein causing diseases related to cancer or neurodegenerative diseases by inhibiting Hsp90, thereby being used in drug medicine and healthy food products for preventing or treating Hsp 90 which is a mediated disease like diseases related to cancer or neurodegenerative diseases.

Description

Flavokawa wine derivatives having Hsp90 inhibitory activity, or pharmaceutically acceptable salts thereof, and medicinal uses thereof,

The present invention relates to a flavor wine derivative having Hsp90 inhibitory activity or a pharmaceutically acceptable salt thereof and a medicinal use thereof.

The Hsp90 protein is one of the most abundant chaperones within eukaryotic cells, and is responsible for the stabilization and regulation of various proteins involved in cell growth differentiation, survival. The substrate protein of Hsp90, called the client protein, contains over 50 cancer-causing proteins. When Hsp90 activity is inhibited, the Hsp90 client proteins are degraded by the proteasome.

Therefore, Hsp90 activity inhibitor can attenuate various cancer-inducing proteins at the same time, and thus has attracted much attention as an anticancer agent that can be applied to a wide variety of cancers. In particular, Hsp90 has been shown to be effective in the treatment of cancer with resistance, since it simultaneously reduces various cancer-inducing proteins.

In addition, it has been reported that Hsp90 inhibitor may be used as a therapeutic agent for degenerative neurological diseases, because proteins that cause degenerative neuropathy are also present in the Hsp90 client proteins.

Korean Patent Publication No. 2007-0038565 discloses 1H-indazol-6-ol compounds and salts thereof as Hsp90 inhibitors.

In the present invention, a new structure of Hsp90 inhibitor is developed, and it is intended to provide a medical use for treating a cancer disease including non-small cell lung cancer or a therapeutic agent for a neurodegenerative disease including Alzheimer's disease.

In order to attain the above object, the present invention provides a flavor wine derivative represented by the following formula (I) or a pharmaceutically acceptable salt thereof:

[Chemical Formula 1]

In Formula 1,

R ₁ to R ₄ may be the same or different and are selected from the group consisting of hydrogen, hydroxy, halogen, C 1 to C 4 alkyl and C 1 to C 4 alkoxy,

R ₁ to R ₄ And the remaining two substituents are each selected from the group consisting of hydrogen, hydroxy, halogen, C1 to C4 alkyl, and C1 to C4 alkoxy. have.

The present invention also provides a pharmaceutical composition for the treatment or prevention of Hsp90 (Heat Shock Protein 90: heat shock protein 90) mediated diseases comprising the flavocaine wine derivative represented by the formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient .

The present invention also provides a health food for preventing or ameliorating Hsp90 (Heat Shock Protein 90: Heat shock protein 90) mediated diseases comprising the flavocain wine derivative represented by the formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient .

The flavocain wine derivatives according to the present invention have excellent inhibitory effect on Hsp90 and induce inhibition of Hsp90 client protein causing cancer disease or degenerative neurological disease through inhibition of Hsp90 and thereby prevent Hsp90 mediated diseases such as cancer diseases or degenerative nerve diseases And can be useful as medicines or health foods for treatment or prevention.

1 is a Western blotting result showing the Hsp90-associated protein inhibitory effect of the flavor wine compound according to the present invention,
FIG. 2 shows the effect of inhibiting H1975 cell proliferation according to the concentration of flavor wine compounds la and li according to the present invention,
FIG. 3 is a Western blotting result showing the Hsp90-related protein inhibitory effect according to the concentration of the flavor wine compound 1i according to the present invention.

The present invention provides a flavor wine compound represented by the following formula (1): < EMI ID =

[Chemical Formula 1]

In Formula 1,

R ₁ to R ₄ may be the same or different and are selected from the group consisting of hydrogen, hydroxy, halogen, C 1 to C 4 alkyl and C 1 to C 4 alkoxy,

R ₁ to R ₄ And the remaining two substituents are each selected from the group consisting of hydrogen, hydroxy, halogen, C1 to C4 alkyl, and C1 to C4 alkoxy. have.

The Plastic Boca wine derivative is the R ₁ to R ₄ may be the same or different each from the above-mentioned formula (I), hydrogen, hydroxy, halogen, C1 to C4 alkyl and C1 to or selected from the group consisting of C4-alkoxy; R ₂ and R ₃ are connected to form a five-membered heterocyclic or benzene ring; and R ₁ and R ₄ are selected from the group consisting of hydrogen, hydroxy, halogen, C 1 to C 4 alkyl and C 1 to C 4 alkoxy; R ₁ and R ₂ are connected to form a benzene ring, and R ₃ and R ₄ are selected from the group consisting of hydrogen, hydroxy, halogen, C 1 to C 4 alkyl and C 1 to C 4 alkoxy.

The flavor wine derivative or a pharmaceutically acceptable salt thereof may be characterized by inhibiting the protein folding reaction by binding to Hsp90 (Heat Shock Protein 90: Thermal Shock Protein 90).

Such pharmaceutically acceptable salts are useful as acid addition salts formed by pharmaceutically acceptable free acids. As the free acid, inorganic acid and organic acid can be used. As the inorganic acid, hydrochloric acid, bromic acid, sulfuric acid, sulfurous acid, phosphoric acid and the like can be used. As the organic acid, citric acid, acetic acid, maleic acid, fumaric acid, , Acetic acid, glycolic acid, succinic acid, tartaric acid, 4-toluenesulfonic acid, galacturonic acid, embonic acid, glutamic acid, citric acid and arpartic acid.

In addition, the flavor wine derivatives of the present invention include not only pharmaceutically acceptable salts, but also all salts, hydrates and solvates which 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 compound of Chemical Formula 1 in a water-miscible organic solvent such as acetone, methanol, ethanol, acetonitrile, etc., And then precipitating or crystallizing the acid solution. Subsequently, in this mixture, a solvent or an excess acid is evaporated and dried to obtain an additional salt, or the precipitated salt may be produced by suction filtration.

The flavocain wine derivative or a pharmaceutically acceptable salt thereof according to the present invention inhibits Hsp90 activity and confirmed that it reduces the cancer-causing proteins such as Her2, EGFR, Met, Akt and Cdk4, and increases Hsp70.

The present invention provides a pharmaceutical composition for the treatment or prevention of Hsp90 (Heat Shock Protein 90: heat shock protein 90) -mediated disease comprising the flavocaine wine derivative represented by the formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

The Hsp90 mediated disease may be a disease selected from the group consisting of cancer diseases, degenerative neurological diseases and viral infections.

The cancer diseases may be selected from the group consisting of non-small cell lung cancer, breast cancer, uterine cancer and pancreatic cancer. Examples of the degenerative neurological diseases include stroke, paralysis, memory loss, memory loss, dementia, forgetfulness, Parkinson's disease, Alzheimer's disease, Pick's disease, Creutzfeld-Kacob disease, Huntington's disease and Lou Gehrig's disease.

In one embodiment of the present invention, the pharmaceutical composition may be formulated with suitable carriers, excipients, disintegrants, sweeteners, coatings, swelling agents, lubricants, lubricants, flavors, antioxidants, buffers , At least one additive selected from the group consisting of a bacteriostatic agent, a diluent, a dispersant, a surfactant, a binder and a lubricant.

Specific examples of carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. Solid formulations for oral administration may be in the form of tablets, pills, powders, granules, capsules These solid preparations can be prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc., into the composition. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, syrups and the like, and various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin which are commonly used simple diluents. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As the suppository base, witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like can be used.

In another embodiment of the present invention, the pharmaceutical composition may be formulated into granules, powders, coated tablets, tablets, pills, capsules, suppositories, gels, syrups, juices, suspensions, emulsions, Can be used.

According to one embodiment of the present invention, the pharmaceutical composition may be administered orally, intraarterally, intraperitoneally, intramuscularly, intraarterally, intraperitoneally, intrasternally, transdermally, nasally, inhaled, topically, rectally, Can be administered to the subject in a conventional manner via the intradermal route.

The preferred dosage of the flavor wine derivative may be varied depending on the condition and body weight of the subject, the type and degree of disease, the type of drug, the route of administration, and the period of time, and may be appropriately selected by those skilled in the art. According to one embodiment of the present invention, the daily dose may be 0.01 to 200 mg / kg, specifically 0.1 to 200 mg / kg, more specifically 0.1 to 100 mg / kg, though it is not limited thereto. The administration may be performed once a day or divided into several times, and thus the scope of the present invention is not limited thereto.

In the present invention, the subject may be a mammal including a human, but is not limited to these examples.

The present invention also provides a health food for preventing or ameliorating Hsp90 (Heat Shock Protein 90: Heat shock protein 90) mediated diseases comprising the flavocaine wine derivative represented by the formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

In one embodiment of the present invention, the health food comprises 0.01 to 90 parts by weight, 0.1 to 90 parts by weight, 1 to 90 parts by weight, or 10 to 90 parts by weight of the flavor wine derivative per 100 parts by weight of the whole health food But are not limited thereto.

In another embodiment of the present invention, the health food may further comprise at least one additive selected from the group consisting of organic acids, phosphates, antioxidants, lactose casein, dextrin, glucose, sugar and sorbitol. The organic acid can be, but is not limited to, citric acid, fumaric acid, adipic acid, lactic acid or malic acid, and the phosphate can be sodium phosphate, potassium phosphate, acid pyrophosphate or polyphosphate (polymeric phosphate) But are not limited to, natural antioxidants such as polyphenols, catechins, alpha-tocopherol, rosemary extract, licorice extract, chitosan, tannic acid or phytic acid.

In another embodiment of the present invention, the health food may contain flavors such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and aging agents (cheese, chocolate, etc.) Organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. In addition, the food composition according to one embodiment of the present invention may contain flesh for the production of natural fruit juice, fruit juice drink and vegetable drink.

According to one embodiment of the invention, the formulation of a health food may be in the form of solid, powder, granule, tablet, capsule, liquid or drink, although not limited thereto.

In addition, the health food includes but is not limited to confectionery, saccharides, ice cream products, dairy products, meat products, fish meat products, tofu or glue, edible oils, noodles, Dried products of ginseng products, kimchi pickles, dried fruits, fruits, vegetables, fruits or vegetables, cutting products, fruit juices, vegetable juices, mixed juice thereof, nuts, noodles, processed livestock products, processed marine products, , Fermented milk food, bean curd food, cereal food, fermented microorganism food, confectionery bakery, condiments, meat processing, acidic beverage, licorice, herb.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited to the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

The following experimental examples are intended to provide experimental examples applicable to the respective embodiments according to the present invention.

< Experimental Example  1> Cell culture

H1975 cells cultured in L- glutamine, streptomycin (500 mg / mL), penicillin (100 units / mL) and 10% fetal bovine serum (FBS) is added 37 ℃ in RPMI 1640 medium, 5% CO ₂ cell incubator Respectively.

< Experimental Example  2> MTS  analysis

It was inoculated by 96 ³ × 10 3 cells per well of a well plate in 100μL medium overnight to attach to the plate Lt; / RTI &gt;

On the next day, the concentrations of the compounds mentioned in the Examples and 1% DMSO control were added to each well, incubated at 37 ° C for 1, 2 and 3 days, and cell viability was measured using Cell Titer 96 Aqueous One Solution Cell Proliferation Assay (Promega) Respectively.

20 μM assay substrate solution was added to each incubated sample. Each sample was incubated at 37 ° C. for 1 hour. The absorbance at 490 nm was measured with a Tecan Infinite F200 Pro plate reader. The results were analyzed by DMSO The percent absorbance of a control group was expressed as a percentage.

< Experimental Example  3> Western Blot  analysis

5 × 10 ⁵ cells per 60 mm culture container were inoculated and cultured overnight so that the cells could be attached to the culture container. In the experimental group, 1a to 1 g of the compounds were added at the concentrations of FIGS. 2 and 3, and 1% DMSO and 1 μM gel Danamycin was added and incubated for an additional 24 hours.

Each of the cells was collected with ice-cold lysis buffer (23 mM Tris-HCl pH 7.6, 130 mM NaCl, 1% NP40, 1% sodium deoxycholate, 0.1% SDS) to give 20 μg of protein per SDS- Loaded, electrophoresed and transferred to a PVDF membrane (Bio-Rad).

The transferred membranes were blocked with 5% skim milk supplemented with TBST and incubated with appropriate concentrations of EGFR, Her2, Met, Akt, Hsp90, Hsp70 and β-Actin primary antibodies and incubated with Horseradish ferro The secondary antibody to which the oxidase was bound was bound and the target protein was identified by ECL chemiluminescence method (Thermo Scientific, USA).

< Experimental Example  4> Chemical properties analysis

All compound synthesis reactions were carried out in the presence of argon in oven-dried glass and all materials purchased were used without further purification.

Thin layer chromatography (TLC) was performed using Merck silica gel 60 F ₂₅₄ plates and TLC plate visualization was performed using UV combination, p -anisaldehyde, seric ammoniomolybdate, ninhydrin and potassium permanganate staining.

NMR spectra were measured using a Bruker 400 (400 MHz for ¹ H; 100 MHz for ¹³ C) spectrometer. The ¹ H and ¹³ C NMR chemical changes were reported per million (ppm) parts compared to TMS, It was used as a reference.

The signal may be a multiplet, a singlet, a doublet, a triplet, a quartet, a broad singlet, a broad doublet, a doublet of doublets, triplets) and dq (doublet of quartets); The connected integers are reported in hertz (Hz).

The synthesized final compound was purified by MPLC (Biotage Isolera One instrument) on a silica gel column (Biotage SNAP HP-Sil).

As a result of the MNR and HPLC (Shimadzu prominence, VP-ODS C18 column) analysis, the compounds used in all experiments were found to have a standard purity of> 95%.

< Example  1> Plavoca wine  Derivatives 1a to 1k synthesis

2,4,6-trihydroxyacetophenone (2) and dimethyl sulfate were reacted with acetone in which potassium carbonate was dissolved to synthesize 2-hydroxy-4,6-dimethoxyacetophenone (3). Flavocain wine derivatives 1a to 1k were synthesized through Claisen-Schmidt condensation reaction with synthesized Compound 3 and aldehydes 4a to 4k in the presence of potassium carbonate.

1k was synthesized in the same manner as in Reaction Scheme 1 below.

1. Synthesis of Compound 1k

[Reaction Scheme 1]

2,4,6-trihydroxyacetophenone (2) and dimethyl sulfate were dissolved in acetone and then reacted to synthesize 2-hydroxy-4,6-dimethoxyacetophenone (3).

A mixture of Compound 3 (0.2 g, 1.02 mmol), 4-methoxy-1-naphthaldehyde (0.19 g, 1.02 mmol) and potassium hydroxide (1.0 g) in 10 mL of methanol was stirred at 50 ° C. for 12 hours .

The mixture was neutralized to pH 6 with 6N HCl and extracted with ethyl acetate. Washing was repeated three times, the organic layer was extracted with saturated NaHCO₃ solution, which was concentrated under reduced pressure after drying with Na ₂ SO _4.

The concentrate was purified by MPLC to give compound 1k as shown in Table 1.

2. Plavoca wine  B (1a) Synthesis

The 1k synthetic procedure was modified to synthesize 1a.

Compound 3 (0.30 g, 1.5 mmol), benzaldehyde (0.18 g, 1.7 mmol) and KOH (0.19 g, 3.4 mmol) were dissolved in 5 mL of methanol and stirred at room temperature for 20 hours.

To the mixture was added 3N HCl to neutralize to pH 6, and the organic layer extracted with ethyl acetate was dried over Na ₂ SO ₄ and concentrated under reduced pressure.

Purification by hexane column chromatography containing 10% ethyl acetate gave 50% yellow solid compound 1a.

^{1 H NMR (400 MHz, CDCl} 3) δ 7.90 (d, J = 15.6Hz, 1H), 7.77 (d, J = 15.6Hz, 1H), 7.61-7.57 (m, 2H), 7.42-7.36 (m, 3H), 6.09 (d, J = 2.4 Hz, 1H), 5.95 (d, J = 2.4 Hz, 1H), 3.90 (s, 3H), 3.82 ESIMS (m / e) = 285 [M + 1] &lt; ⁺ &gt;.

3. Synthesis of Compound 1b

Was synthesized in the same manner as Compound 1k, except that the kind of aldehyde was changed.

35% yield. _Rf = 0.29 (3: 7 ethyl acetate: hexane). ^{1 H NMR (400 MHz, Acetone} ) δ 7.91 (d, J = 15.2Hz, 1H), 7.80 (d, J = 15.6Hz, 1H), 7.65 (d, J = 8.8Hz, 2H), 6.95 (d, J = 8.4 Hz, 2H), 6.16 (d, J = 2.4 Hz, 1H), 6.12 (d, J = 2.0 Hz, 1H) 4.04 (s, 3H), 3.91 (s, ^{13 C} NMR (100 MHz, DMSO) δ 193.3, 166.6, 166.3, 162.9, 144.4, 131.7, 129.2, 124.8, 117.1, 107.3, 94.9, 92.1, 57.2, 56.07

4. Synthesis of Compound 1c

Was synthesized in the same manner as Compound 1k, except that the kind of aldehyde was changed.

58% yield. _Rf = 0.28 (2: 8 ethyl acetate: hexane).

^{1 H NMR (400 MHz, CDCl} 3) δ 14.41 (s, 1H), 7.79 (d, J = 1.2Hz, 2H), 7.56 (d, J = 8.8Hz, 2H), 6.92 (d, J = 8.4Hz 2H), 6.10 (d, J = 2.0Hz, 1H), 5.95 (d, J = 2.4Hz, 1H), 3.91 (s, 3H), 3.85 (s, 3H), 3.83 (s, ^{13 C} NMR (100 MHz, CDCl ₃ ) δ 192.9, 168.7, 166.3, 162.8, 161.7, 142.8, 130.4, 128.6, 125.4, 114.7, 106.6, 94.1, 91.5, 56.1, 55.9, 55.7

5. Synthesis of Compound 1d

Was synthesized in the same manner as Compound 1k, except that the kind of aldehyde was changed.

21% yield. _Rf = 0.14 (2: 8 ethyl acetate: hexane).

^{1 H NMR (400 MHz, CDCl} 3) δ14.41 (s, 1H), 7.80 (d, J = 15.6Hz, 1H), 7.75 (d, J = 15.6Hz, 1H), 7.21 (dd, J = 8.2 Hz, J = 2.0Hz, 1H) , 7.12 (d, J = 1.6Hz, 1H), 6.89 (d, J = 8.4Hz, 1H), 6.11 (d, J = 2.4Hz, 1H), 5.96 (d, J = 2.0 Hz, 1H), 3.94 (s, 3H), 3.93 (s, 3H), 3.91 (s, 3H), 3.83 (s, 3H). ^{13 C} NMR (100 MHz, CDCl ₃ ) δ 192.8, 168.7, 166.4, 162.7, 151.4, 149.5, 143.0, 128.9, 125.8, 123.0, 111.5, 110.8, 106.7, 94.2, 91.6, 56.3, 56.2, 56.1, 55.9

6. Synthesis of Compound 1e

Was synthesized in the same manner as Compound 1k, except that the kind of aldehyde was changed.

69% yield. _Rf = 0.17 (1: 9 ethyl acetate: hexane).

^{1 H NMR (400 MHz, CDCl} 3) δ 14.39 (s, 1H), 7.73 (d, J = 2.8Hz, 2H), 7.11-7.07 (m, 2H), 6.83 (d, J = 8.0Hz, 1H) , 6.09 (d, J = 2.4Hz , 1H), 6.01 (s, 2H), 5.95 (d, J = 2.0Hz, 1H), 3.90 (s, 3H), 3.82 (s, 3H), 1.25 (s, 3H). ¹³ C NMR (100 MHz, CDCl ₃ ) δ 192.8, 168.7, 166.4, 162.7, 149.9, 148.6, 142.7, 130.3, 125.8, 125.4, 108.9, 106.9, 106.6, 101.9, 94.1, 91.5, 56.2, 55.9

7. Compound 1f Synthesis

Was synthesized in the same manner as Compound 1k, except that the kind of aldehyde was changed.

23% yield. _Rf = 0.18 (2: 8 ethyl acetate: hexane).

^{1 H NMR (400 MHz, CDCl} 3) δ 14.32 (s, 1H), 7.80 (d, J = 15.6Hz, 1H), 7.70 (d, J = 15.2Hz, 1H), 6.84 (s, 2H), 6.11 (d, J = 2.4Hz, 1H ), 5.96 (d, J = 2.0Hz, 1H), 3.91 (s, 6H), 3.91 (s, 3H), 3.90 (s, 3H), 3.84 (s, 3H) . ¹³ CNMR (100 MHz, CDCl ₃ ) δ 192.7, 168.8, 166.5, 162.7, 153.7, 142.8, 140.4, 131.5, 127.3, 106.6, 105.9, 94.2, 91.7, 61.4, 56.5, 56.2, 56.0

8. Compound 1g Synthesis

Was synthesized in the same manner as Compound 1k, except that the kind of aldehyde was changed.

20% yield. _Rf = 0.24 (2: 8 ethyl acetate: hexane).

^{1 H NMR (400 MHz, CDCl} 3) δ 14.25 (d, J = 0.4Hz, 1H), 8.08 (d, J = 15.6Hz, 1H), 7.77 (d, J = 15.6Hz, 1H), 6.98 (s , 1H), 6.08 (d, J = 2.4Hz, 1H), 5.92 (d, J = 2.0Hz, 1H), 3.93 (s, 3H), 3.91 (s, 3H), 3.90 (s, 3H), 3.87 (s, 3 H), 3.81 (s, 3 H). ^{13 C NMR (100 MHz, CDCl} 3) δ162.4, 168.8, 166.4, 162.7, 152.3, 150.5, 145.2, 138.4, 129.6, 129.5, 122.7, 106.5, 106.2, 94.1, 91.6, 61.6, 61.4, 56.4, 56.1, 55.9

9. Synthesis of Compound 1h

Was synthesized in the same manner as Compound 1k, except that the kind of aldehyde was changed.

15% yield. _Rf = 0.16 (2: 8 ethyl acetate: hexane). ^{1 H NMR (400 MHz, CDCl} 3) δ 14.24 (s, 1H), 8.08 (d, J = 15.2Hz, 1H), 7.72 (d, J = 15.6Hz, 1H), 7.01 (s, 1H), 6.09 (d, J = 2.4Hz, 1H ), 5.94 (d, J = 2.4Hz, 1H), 3.93 (s, 3H), 3.92 (s, 3H), 3.90 (s, 3H), 3.88 (s, 3H) , &Lt; / RTI &gt; 3.82 (s, 3H). ^{13 C NMR (100 MHz, CDCl} 3) δ192.4, 168.8, 166.7, 162.7, 153.0, 151.5, 145.0, 141.0, 131.4, 129.8, 113.7, 106.7, 106.6, 94.2, 91.6, 61.7, 61.3, 56.4, 56.1, 55.9

10. Synthesis of Compound 1i

Was synthesized in the same manner as Compound 1k, except that the kind of aldehyde was changed.

32% yield. _Rf = 0.27 (1: 9 ethyl acetate: hexane). ^{1 H NMR (400 MHz, CDCl} 3) δ 14.40 (s, 1H), 8.61 (d, J = 15.6Hz, 1H), 8.31 (d, J = 8.4Hz, 1H), 7.96 (d, J = 15.2Hz , 1H), 7.90-7.87 (m, 2H), 7.83 (d, J = 7.2Hz, 1H), 7.60-7.49 (m, 3H), 6.13 (d, J = 2.0,1H), 5.96 (d, J = 2.4 Hz, 1 H), 3.89 (s, 3 H), 3.82 (s, 3 H). ^{13 C NMR (100 MHz, CDCl} 3) δ192.8, 168.7, 166.6, 162.8, 139.3, 134.0, 133.2, 132.0, 130.6, 130.4, 129.0, 127.0, 126.4, 125.7, 125.4, 123.9, 106.6, 94.1, 91.5, 57.4, 56.1

11. Compound 1j Synthesis

Was synthesized in the same manner as Compound 1k, except that the kind of aldehyde was changed.

70% yield. _Rf = 0.21 (1: 9 ethyl acetate: hexane). ^{1 H NMR (400 MHz, CDCl} 3) δ 14.40 (s, 1H), 7.98 (t, J = 6.8Hz, J = 5.2Hz, 3H), 7.88-7.82 (m, 3H), 7.52-7.50 (m, 3H), 6.12 (d, J = 2.4 Hz, 1H), 5.97 (d, J = 2.4 Hz, 1H), 3.94 (s, 3H), 3.83 (s, ^{13 C} NMR (100 MHz, Acetone)? 193.4, 169.3, 167.7, 163.9, 143.2, 135.3, 134.6, 134.1, 131.4, 129.7, 129.6, 128.7, 128.2, 127.7, 124.8, 107.0, 94.8, 92.0, 56.7, 56.2

< Example  2> Plavoca wine  Identification of biological effects of derivatives

One. in vitro in Plavoca wine  Derivative Antiproliferative activity  Confirm

The antiproliferative activity of flavone wine derivatives (1a-k) synthesized in H1975, an in vitro model of gefitinib-resistant non-small lung cancer species (NSCLC), was confirmed.

H1975 cells were treated with each compound (1a-k) at a concentration of 30 μM for 72 hours.

The inhibitory activity of each compound on H1975 cell growth was measured by MTS colorimetric assay and expressed as% inhibition compared with DMSO control.

As a result, as shown in Table 1, inhibition rates of the compounds 1a, 1i and 1j were 34, 36 and 30%, respectively, and the inhibition of H1975 cell proliferation was the best.

2. Plavoca wine  Derivative Hsp90  Confirmation of inhibition

In order to confirm whether the antiproliferative effect of flavone wine derivatives (1a-k) is related to Hsp90 inhibition, H1975 cells were treated with 30 μM of each compound for 24 hours, and the Hsp90 client proteins EGFR, Met, Her2, Akt And Cdk 4 expression were analyzed by Western blotting, and geldanamycin (GA, 1 mM) and DMSO (D) were used as negative control and positive control, respectively, and Hsp70, Hsp90 and β-actin were also identified.

As a result, as shown in FIG. 1, strong degradation of EGFR, Met, Her2, Akt and Cdk4 and induction of important Hsp70 were confirmed in the test group treated with the compound 1a and 1i. From these results, it was confirmed that 1a and 1i interfere with the protein chaperoning function of Hsp90.

It is also noteworthy that the naphthalene group, which is a B-ring of 1j and 1k, has been shown to reduce the expression of EGFR, Met and Her2, the tyrosine kinase (RTK) protein receptors of cell membranes, but the effect on Akt, Cdk 4 and Hsp70 proteins It was not big.

3. Plavoca wine  Compounds 1a and 1i by concentration and time Antiproliferative activity  Confirm

From these results, MTS analysis was performed to confirm the antiproliferative activity of the compounds la and l in H1975 cells.

H1975 cells were treated with the compounds 1a and 1i at the same concentrations as in FIG. 3, and the cell proliferation was measured every 1, 2, and 3 days by the MTS method, and the results were shown as ± SD (n = 4) values.

As a result, it was confirmed that the compounds 1a and 1i inhibited H1975 cell proliferation in a concentration and time-dependent manner as shown in Fig.

4. Compound 1i Hsp90  Confirmation of inhibition

As shown in Fig. 1 and Fig. 2, compound 1i showed better Hsp90 inhibitory effect than 1a and optimal concentration of compound 1i for Hsp90 inhibition was measured.

H1975 cells were treated with compound 1i at 0, 5, 10, 20, 30 and 50 μM for 24 hours and the expression levels of Met, Her2, Akt, Cdk4, Hsp70, Hsp90 and β-actin were measured by Western blotting.

4, Compound 1i decreased the expression of the Hsp90 client proteins Met, Her2, Akt and Cdk4, while Hsp70 and Hsp90 increased in a concentration-dependent manner.

On the other hand, there was no change in β-actin, an Hsp90-independent protein.

In addition, Her2 protein was completely inhibited from 20 μM concentration of Compound 1i, and Met, Akt and Cdk 4 were significantly decreased from 30 μM concentration of 1i. From the results, it was found that Her2 protein was more effective than Compound 1i The most sensitive reaction was observed.

R ₁ R ₂ R ₃ R ₄  Yield compound Percent inhibition% ^d One H H H H 50 1a ^a 34 2 H H OH H 35 1b ^b 15 3 H H OMe H 58 1c ^c 11 4 H OMe OMe H 21 1d 5 5 H OCH ₂ O H 69 1e 15 6 H OMe OMe OMe 23 1f 17 7 Cl OMe OMe OMe 20 1g 10 8 Br OMe OMe OMe 15 1h 13 9 CHCHCHCH H H 32 1i 36 10 H CHCHCHCH H 70 1j 30 11 CHCHCHCH OMe H 18 1k 22

^a Flavor wine B, ^b Praloca wine C, ^c Flavor wine A

^d compound 30μM processed with H1975 cell proliferation inhibition rate (%)

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that such detail is solved by the person skilled in the art without departing from the scope of the invention. will be. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (8)

A flavor wine derivative represented by the following formula (1): &lt; EMI ID =
[Chemical Formula 1]

In Formula 1,
R 1 to R 4 may be the same or different and are selected from the group consisting of hydrogen, hydroxy, halogen, C 1 to C 4 alkyl and C 1 to C 4 alkoxy;
Two substituents R &lt; 1 &gt; to R &lt; 4 &gt; are connected to form a 5- or 6-membered ring, and the remaining two substituents are each selected from the group consisting of hydrogen, hydroxy, halogen, C1 to C4 alkyl and C1 to C4 alkoxy. The method of claim 2,
The Plastic Boca wine derivative is the R ₁ to R ₄ may be the same or different each from the above-mentioned formula (I), hydrogen, hydroxy, halogen, C1 to C4 alkyl and C1 to or selected from the group consisting of C4-alkoxy; R ₂ and R ₃ are connected to form a five-membered heterocyclic or benzene ring; and R ₁ and R ₄ are selected from the group consisting of hydrogen, hydroxy, halogen, C 1 to C 4 alkyl and C 1 to C 4 alkoxy; Wherein R ₁ and R ₂ are connected to form a benzene ring and R ₃ and R ₄ are selected from the group consisting of hydrogen, hydroxy, halogen, C 1 to C 4 alkyl and C 1 to C 4 alkoxy. A wine derivative or a pharmaceutically acceptable salt thereof. The method according to claim 1 or 2,
Wherein the flavocain wine derivative or a pharmaceutically acceptable salt thereof is characterized in that it is combined with Hsp90 (Heat Shock Protein 90: Thermal Shock Protein 90) to inhibit the protein folding reaction. The flavocain wine derivative or a pharmaceutically acceptable salt thereof, salt. A pharmaceutical composition for treating or preventing Hsp90 (Heat Shock Protein 90: heat shock protein 90) -mediated disease, which contains, as an active ingredient, a flavor wine derivative represented by the following formula (1) or a pharmaceutically acceptable salt thereof.
[Chemical Formula 1]

In Formula 1,
R 1 to R 4 may be the same or different and are selected from the group consisting of hydrogen, hydroxy, halogen, C 1 to C 4 alkyl and C 1 to C 4 alkoxy;
Two substituents of R 1 to R 4 are connected to form a 5-or 6-membered ring, and the remaining two substituents are selected from the group consisting of hydrogen, hydroxy, halogen, C 1 to C 4 alkyl and C 1 to C 4 alkoxy. The method of claim 4,
Wherein the Hsp90 mediated disease is selected from the group consisting of cancer diseases, degenerative neurological diseases and viral infections. 9. The pharmaceutical composition for treating or preventing Hsp90 (Heat Shock Protein 90: Heat shock protein 90) -mediated disease. The method of claim 5,
Wherein said cancer is selected from the group consisting of non-small cell lung cancer, breast cancer, uterine cancer and pancreatic cancer. The method of claim 5,
The degenerative neurological disease is selected from the group consisting of stroke, stroke, memory loss, memory impairment, cognitive dementia, amnesia, Alzheimer's disease, Parkinson's disease, peak disease, Creutzfeldt-Jakob disease, Huntington's disease and Lou Gehrig's disease (Heat Shock Protein 90: Thermal Shock Protein 90) mediated disease. A health food for preventing or ameliorating Hsp90 (Heat Shock Protein 90: Heat shock protein 90) -mediated disease, which contains, as an active ingredient, a flavor wine derivative represented by the following formula (1) or a pharmaceutically acceptable salt thereof.
[Chemical Formula 1]

In Formula 1,
R 1 to R 4 may be the same or different and are selected from the group consisting of hydrogen, hydroxy, halogen, C 1 to C 4 alkyl and C 1 to C 4 alkoxy;
Two substituents of R 1 to R 4 are connected to form a 5-or 6-membered ring, and the remaining two substituents are selected from the group consisting of hydrogen, hydroxy, halogen, C 1 to C 4 alkyl and C 1 to C 4 alkoxy.

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