KR20190014474A - New dihydroxyphenyl stereoisomer having inhibitory activity on Hsp90 and medical use thereof - Google Patents

Abstract

The present invention relates to a novel dihydroxyphenyl-based stereoisomer having Hsp90-inhibiting activity and a pharmaceutical composition or a health functional food comprising the same. The stereoisomer can be confirmed to have an excellent Hsp90 inhibitory effect of 30 times or more, which is unpredictable from a racemic mixture, thereby being usefully used as a pharmaceutical composition for preventing or treating Hsp90-mediated disease selected from a group consisting of cancerous disease, neurodegenerative disease, and viral infection or used as a health functional food for preventing or alleviating Hsp90-mediated disease selected from the same.

Description

[0001] The present invention relates to dihydroxyphenyl-based stereoisomers having HSP90 inhibitory activity and medicinal uses thereof,

The present invention relates to dihydroxyphenyl-based stereoisomers having HSP90 inhibitory activity and their medical uses.

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.

The Hsp90 inhibitor started from the development of the natural substance geldanamycin (GA). GA has been shown to induce the degradation of the client protein Src through inhibition of Hsp90 in 1994. Since then, inhibitors targeting Hsp90 have been actively developed. However, GA has a strong anticancer effect, but has problems of liver toxicity, solubility and stability. In order to compensate for this, the GA derivatives, Tanespimycin (17-AAG), alvespimycin (17-DMAG) and retaspimycin have been developed but their structural characteristics have not solved the problem. Since then, research on Hsp90 inhibitors of various structures has been conducted at the clinical stage, but since FDA-approved drugs have not yet been developed, it is necessary to develop new, more potent compounds.

Korean Patent Laid-Open No. 10-2008-0002809 (published on January 4, 2008)

It is an object of the present invention to provide a novel dihydroxyphenyl based stereoisomer.

It is another object of the present invention to provide a pharmaceutical composition for preventing or treating Hsp90-mediated diseases, which comprises a novel dihydroxyphenyl-based stereoisomer as an active ingredient.

It is another object of the present invention to provide a health functional food for preventing or ameliorating Hsp90-mediated diseases, which comprises a novel dihydroxyphenyl-based stereoisomer as an active ingredient.

The present invention provides dihydroxyphenyl-based stereoisomers represented by the following formula (1).

[Chemical Formula 1]

In Formula 1,

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

R ₂ is any one selected from the group consisting of C 3 -C 6 cycloalkyl, phenyl, halogen, C 1 -C 4 alkyl and C 1 -C 4 alkoxy,

R ₃ is C 1 -C 4 alkyl or C 1 -C 4 alkoxy.

The present invention provides a pharmaceutical composition for preventing or treating Heat Shock Protein 90 (Hsp90) -mediated disease, which comprises the dihydroxyphenyl stereoisomer represented by Formula 1 as an active ingredient.

The present invention also provides a health food for preventing or ameliorating a heat shock protein 90 (Hsp90) mediated disease comprising the dihydroxyphenyl stereoisomer represented by the above formula (1) as an active ingredient.

The present invention relates to a dihydroxyphenyl-based stereoisomer having Hsp90 inhibitory activity, and a pharmaceutical composition or health functional food comprising the same, wherein the stereoisomer is excellent in an unexpectedly superior 30 times or more The Hsp90 inhibitory effect can be confirmed. Therefore, it can be effectively used as a pharmaceutical composition for preventing or treating Hsp90-mediated diseases selected from the group consisting of cancer diseases, degenerative neurological diseases and viral infections, or health functional foods for prevention or improvement.

Figure 1 shows the chiral HPLC separation results to identify the stereoisomers of Compound 17.
FIG. 2 shows the effect of inhibiting H1975 cell proliferation through the regulation of Hsp90 client protein of the compound 17 stereoisomer, Peak 1, wherein FIG. 2 (A) is the MTS analysis result and FIG. 2 (B) is the Western blot analysis result.

Hereinafter, the present invention will be described in more detail.

The inventors of the present invention have found that when a compound showing heat shock protein 90 (Heat Shock Protein 90, Hsp90) inhibitory effect is produced, the stereoisomer represented by the following formula 1 has an excellent Hsp90 inhibitory effect To complete the present invention.

The present invention can provide a dihydroxyphenyl-based stereoisomer represented by the following formula (1): < EMI ID =

[Chemical Formula 1]

In Formula 1,

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

R ₂ is any one selected from the group consisting of C 3 -C 6 cycloalkyl, phenyl, halogen, C 1 -C 4 alkyl and C 1 -C 4 alkoxy,

R ₃ is C 1 -C 4 alkyl or C 1 -C 4 alkoxy.

More specifically, the stereoisomer may be a dihydroxyphenyl-based stereoisomer represented by the following general formula (2).

(2)

The present invention can provide a pharmaceutical composition for preventing or treating Heat Shock Protein 90 (Hsp90) -mediated disease comprising dihydroxyphenyl based stereoisomer represented by the following formula (1) as an active ingredient:

[Chemical Formula 1]

In Formula 1,

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

R ₂ is any one selected from the group consisting of C 3 -C 6 cycloalkyl, phenyl, halogen, C 1 -C 4 alkyl and C 1 -C 4 alkoxy,

R ₃ is C 1 -C 4 alkyl or C 1 -C 4 alkoxy.

More specifically, the stereoisomer may be a compound represented by the following formula (2). The pharmaceutical composition may be a pharmaceutical composition for preventing or treating Heat Shock Protein 90 (Hsp90) mediated diseases.

(2)

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

More specifically, the cancer diseases include cancer of the non-small cell lung, breast, ovarian, uterine, pancreatic, lung, stomach, liver, But are not limited to, cancer, rectal cancer, acute myelogenous leukemia, chronic myelogenous leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, and blood cancer.

In addition, the degenerative neurological disorder may be selected from the group consisting of stroke, paralysis, memory loss, memory impairment, dementia, forgetfulness, Parkinson's disease, Alzheimer's disease, Pick's disease, Creutzfeld-Kacob disease, Huntington's disease, But the present invention is not limited thereto.

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

Examples of 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, Methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil.

The pharmaceutical composition according to the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions according to a conventional method .

In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose sucrose), lactose, gelatin, and the like.

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, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included .

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. 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. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

Further, the dosage of the pharmaceutical composition according to the present invention may be increased or decreased depending on the route of administration, degree of disease, sex, weight, age, and the like. Thus, the dosage amounts are not intended to limit the scope of the invention in any manner.

The pharmaceutical composition may be administered to mammals such as rats, mice, livestock, humans, and the like in a variety of routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intratracheal, intrauterine or intracerebroventricular injections.

In addition, the present invention provides a health functional food for preventing or ameliorating a heat shock protein 90 (Hsp90) mediated disease comprising the dihydroxyphenyl based stereoisomer represented by the above formula (1) or (2) as an active ingredient.

The health functional food may be provided in the form of powder, granules, tablets, capsules, syrups or beverages. The health functional food may be used together with other food or food additives other than the compound represented by the formula (1) Can be suitably used according to the method of The amount of the active ingredient to be mixed can be suitably determined according to its use purpose, for example, prevention, health or therapeutic treatment.

The stereoisomers represented by the above formula (1) or (2) contained in the health functional food may be used in accordance with the effective dose of the pharmaceutical composition, but may be used for health and hygiene purposes or for long- It may be less than the above range, and since the active ingredient has no problem in terms of safety, it can be used in an amount exceeding the above range.

There is no particular limitation on the kind of the above health functional food and examples thereof include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, , Drinks, alcoholic beverages and vitamin complexes.

In addition, the dihydroxyphenyl-based stereoisomers 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, the dihydroxyphenyl stereoisomer of the formula (1) or (2) is dissolved in a water-miscible organic solvent such as acetone, methanol, ethanol, or acetonitrile Or the like, adding an excess amount of an organic acid, or adding an aqueous acid solution of an inorganic acid, followed by precipitation or crystallization. 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.

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.

< Reference example > Laboratory equipment

All reagents and solvents were purchased from the manufacturer and used without further purification.

All experiments dealing with moisture sensitive compounds were performed under an argon atmosphere.

Concentration or solvent removal was carried out using a rotary evaporator under reduced pressure.

Analytical thin layer chromatography was performed on precoated silica gel F ₂₅₄ TLC plates (silica gel F254 TLC plates, E, Merck) and visualized by staining UV light with iodine gas.

Column chromatography is carried out on medium (medium pressure) on silica (Merck Silica gel 40-63 μm) or in a biotage SP1 flash purification system (Biotage SP1) using prepacked silica gel cartridges (Biotage) flash purification system).

NMR analysis was carried out using ARX-300 (300 MHz or more) manufactured by Bruker.

Chemical shifts were recorded in per million (δ). The deuterium lock signal of the sample solvent was used as a reference, and the coupling constants (J) were recorded in hertz (Hz).

The splitting pattern abbreviations are: s, singlet; d, doublet; t, triplet; q, quartet, dd, doublet of doublets; m, multiplet.

<Experimental Example>

1. Cell culture

Sk-Br3 (Korean cell line bank), a breast cancer cell, and H1975 cell (ATCC), a non-small cell lung cancer, were treated with 25 mM HEPES [4- (2-hydroxyethyl) -1-piperazineethanesulfonic acid] (Streptomycin, 500 mg / mL), penicillin (100 units / mL) and 10% fetal bovine serum (L-glutamine) fetal bovine serum, FBS).

The cells were cultured in a humidified atmosphere of 37 ° C and 5% CO ₂ .

2. Cell proliferation assay

H1975 cells were dispensed into a 96-well plate to 1500 cells per well, added to a volume of 100 μL, and then the cells were allowed to adhere overnight.

The following day, each compound was added to the wells at various concentrations and then the cells were incubated at 37 占 for 3 days.

Cell viability was determined using the Promega Cell Titer 96 Aqueous One Solution cell proliferation assay.

After incubation with compound, 20 μL of the assay substrate solution was added to each well and incubated at 37 ° C. for 1 hour. The absorbance at 490 nm was measured on a Tecan Infinite F200 Pro plate reader and the percentage of absorbance of the incubated cells in DMSO Cell viability.

3. Western Blot  analysis

H1975 cells were plated at a density of 1x10 &lt; ⁶ &gt; / dish in a 100 mm culture dish and adhered overnight. Each compound was added to the cells at a concentration of 0.1 and 0.5 μM and the cells were further incubated for 24 hours. Cells were then harvested with cold lysis buffer (23 mM Tris-HCl pH 7.6, 130 mM NaCl, 1% NP-40, 1% sodium deoxycholate, 0.1% SDS) and 30 μg lysine / PAGE, and then transferred to a PVDF membrane (Bio-Rad).

The membranes were blocked with TBST containing 5% skim milk and incubated with the appropriate antibodies (EGFR, p-EGFR, Her2, Met, Akt, Hsp90, Hsp70, and beta-Actin).

After binding appropriate secondary antibodies conjugated to horseradish peroxidase, proteins were visualized by ECL chemiluminescence according to the manufacturer's instructions (GE healthcare, USA).

4. Fluorescence polarization assay

All fluorescence polarization analyzes (FP) were performed on a black, round bottom 96-well plate using a microplate reader.

For FP analysis, HFB buffer (20 mM HEPES pH 7.3, 50 mM KCl, 5 mM MgCl ₂ , 20 mM Na ₂ MoO ₄ , 0.01% NP-40), 30 nM recombinant Hsp90 alpha full length protein, 5 nM (GA-FITC) inhibitor labeled with fluorescein isothiocyanate, 0.1 mg / mL bovine serum globulin (BGG), 2 mM 1,4-dithiothreitol (DTT) and various concentrations of compound Was added.

The HFB buffer in all wells was brought to a final volume of 100 μL. Plates were incubated at 4 ° C for 16 hours and the polarization values of millipolarization units were measured at an emission wavelength of 495 nm and a radiation wavelength of 530 nm.

All experimental data were analyzed using Prism software (version 5.0, Graphpad Software, San Diego, Calif.).

5. Chirality HPLC  detach

Chiral HPLC separation of Compound 17 was performed using Daicel chiral technologies (china) CO. .

< Example  1> Dihydroxyphenyl series Stereoisomer Compound 17 Synthesis

[Reaction Scheme 1]

Compound 17 as an isopropyl inhibitor was synthesized in the same manner as in Scheme 1.

First, the Friedel-Crafts alkylation of methyl 2,4-dihydroxybenzoate (6) was carried out with isopropyl bromide and aluminum chloride.

Compound 12 was protected with aryl bromide in the presence of potassium carbonate to give ester 13, which was converted using sodium hydroxide to give carboxylic acid 14 in 58% yield.

The amide coupling reaction of carboxylic acid 14 with 1-methyl-3-phenylpiperazine (15) was carried out in the presence of DIPEA in EDC, HOBt or DMF to give amide 16.

Finally, the aryl-protected group was removed using PdCl ₂ (PPh ₃ ) ₂ and ammonium formate under microwave irradiation to give compound 17.

Chiral HPLC separation of compound 17 was performed using a chiral HPLC column in hexane and ethanol eluants, and as a result, single optical isomeric peaks 1 and 2 were identified as in FIG.

1-1. methyl  2,4- Dihydroxybenzoate  (Methyl 2,4- dihydroxybenzoate ; 6)

MeOH (40 ml) to which 2,4-dihydroxybenzoic acid (10.2 g, 66.0 mmol) and sulfuric acid (5 ml) were added in a reflux condenser under argon was stirred at 100 ° C for 12 hours.

The mixture was cooled to room temperature and concentrated under reduced pressure, then 40 mL of H ₂ O was poured in a water bath. The white solid formed by the above procedure was dissolved in ethyl acetate for filtration and washed with saturated NaHCO ₃ solution.

The organic layer was dried over Na ₂ SO ₄ and concentrated under reduced pressure to obtain Compound 6 in 86% yield. Rf = 0.20 (2: 8 ethyl acetate: hexane). ^{1 H NMR (400 MHz, CDCl} 3) δ 10.97 (s, 1H), 7.74 (d, J = 8.8 Hz, 1H), 6.40 (d, J = 2.0 Hz, 1H), 6.37 (dd, J = 8.8 Hz , 2.4 Hz, 1 H), 5.36 (s, 1 H), 3.91 (s, 1 H).

1-2. methyl  2,4- Dihydroxy -5- Isopropylbenzoate  (methyl 2,4-dihydroxy-5-isopropylbenzoate; 12)

Compound 6 (3.9 g, 23.0 mmol), 2-bromopropane (4.3 mL, 46.0 mmol) and aluminum chloride (6.1 g, 46.0 mmol) were added to a CH ₂ Cl ₂ solution, And stirred in a reflux condenser at 50 &lt; 0 &gt; C for 24 hours. 2-Bromopropane (4.3 mL, 46.0 mmol) was added to the reaction mixture three times at 6 hour intervals.

The mixture was neutralized to pH 5 with 10% NaOH, concentrated under reduced pressure, and extracted with ethyl acetate. The organic layer was washed three times with saturated NaHCO ₃ solution, dried over Na ₂ SO ₄ , concentrated under reduced pressure and purified by column to give compound 12 in 45% yield. Rf = 0.21 (1: 4 ethyl acetate: hexane). ¹ H NMR (400 MHz, CDCl ₃ )? 10.8 (s, IH), 7.64 (s, IH), 6.34 (s, IH), 5.53 m, 1 H), 1.25 (d, J = 10.8 Hz, 6H). ¹³ C NMR (100 MHz, CDCl 3)? 170.7, 161.6, 159.6, 128.1, 127.1, 105.7, 103.2, 52.2, 26.7, 22.8

1-3. methyl  2,4- Bis ( Allyloxy ) -5- Isopropylbenzoate  (methyl 2,4-bis (allyloxy) -5-isopropylbenzoate; 13)

A mixture of compound 12 (2.1 g, 10.3 mmol), allyl bromide (2.3 mL, 26.8 mmol) and potassium carbonate (3.7 g, 26.8 mmol) dissolved in DMF was stirred at room temperature for 18 hours, The mixture was dissolved in ethyl acetate.

The organic layer was washed with H ₂ O, dried over Na ₂ SO ₄ and concentrated under reduced pressure to give Compound 13 in 85% yield.

^{1 H NMR (400 MHz, CDCl} 3): δ 7.71 (s, 1H), 6.41 (s, 1H), 6.10-5.97 (m, 2H), 5.50 (dd, J = 17.2 Hz, 1.6 Hz, 1H), (Dd, J = 9.6 Hz, 4.8 Hz, 4H), 3.84 (s, 1H), 3.26 (d, J = -3.19 (m, 1 H), 1.19 (d, J = 6.8 Hz, 6 H).

1-4. 2,4- Bis ( Allyloxy ) -5- Isopropylbenzoic acid  (2,4-bis ( allyloxy ) -5-isopropylbenzoic acid; 14)

30 mL of MeOH 30 mL-H ₂ O in which compound 13 (2.5 g, 8.6 mmol) and sodium hydroxide (1.7 g, 43.1 mmol) was dissolved was stirred at room temperature for 24 hours.

The mixture was dissolved in ethyl acetate. The organic layer was washed with 3N-HCl solution, concentrated under reduced pressure, and purified by column to obtain Compound 14 in 58% yield. Rf = 0.18 (1: 4 ethyl acetate: hexane). ^{1 H NMR (400 MHz, CDCl} 3): δ 7.97 (s, 1H), 6.43 (s, 1H), 6.10-5.97 (m, 2H), 5.47-5.39 (m, 2H), 5.30 (d, J = (M, 1H), 1.18 (d, J = 6.8 Hz, 6H), 4.7 (d, J = 5.6 Hz, 2H) .

1-5. (2,4- Bis ( Allyloxy ) -5- Isopropylphenyl )(4- methyl -2- Phenylpiperazine -1-yl) methanone [(2,4-bis ( allyloxy ) -5- isopropylphenyl ) (4-methyl-2- phenylpiperazine -1-yl) methanone; 16]

(0.18 g, 0.66 mmol), 1-methyl-3-phenylpiperazine (0.18 g, 0.99 mmol), 1-ethyl-3- (3- dimethylaminopropyl) [1-ethyl-3- (3-dimethylaminopropyl); (0.09 g, 0.66 mmol) and N, N-diisopropylethylamine (0.09 mL, 0.66 mmol) were added to a solution of 1-hydroxybenzotriazole Was stirred at 120 &lt; 0 &gt; C for 3 hours under microwave irradiation (Biotage Initiator).

The mixture was dissolved in ethyl acetate and the organic layer was washed with 1N HCl solution, dried over Na ₂ SO ₄ , concentrated under reduced pressure and purified by MPLC (Biotage SNAP HP-Sil column) to give Compound 17 in 92% yield . Rf = 0.24 (3: 7 ethyl acetate: hexane).

1-6. (2,4- Dihydroxy -5- Isopropylphenyl )(4- methyl -2- Phenylpiperazine -1 day) Methanone  [(2,4- dihydroxy -5- isopropylphenyl ) (4-methyl-2- phenylpiperazine -1-yl) methanone; 17]

_{Pd Cl 2 (PPh 3) 2} (10 mg) and ammonium formate (ammonium formate; 227 mg) while the addition of 16 (0.26 g, 0.61 mmol) compound in THF 4 mL for the present and a 30-minute 120 ℃ under microwave irradiation Lt; / RTI &gt;

The reaction mixture was diluted with ethyl acetate and the organic layer was washed with water and dried over Na ₂ SO _4. Concentration under reduced pressure and purification by MPLC gave Compound 17 in two steps. Rf = 0.32 (8: 2 ethyl acetate: hexane). ^{1 H NMR (400 MHz, CDCl} 3): 7.45 (d, J = 7.6 Hz, 2H), 7.37 (t, J = 7.2 Hz, 7.6 Hz, 2H), 7.25 (t, J = 7.8 Hz, 7.6 Hz, (T, 1H), 6.99 (s, 1H), 6.40 (s, 1H), 5.58 J = 12.4 Hz, 10.4 Hz, 1H), 3.05-3.00 (m, 1H), 2.79 (d, J = 10.8 Hz, 1H), 2.47 , 3H), 2.22-2.16 (m, 1H), 0.95 (dd, J = 20.0 Hz, 6.0 Hz, 6H). ^{13 C NMR (100 MHz, CDCl} 3) δ 171.5, 158.1, 138.8, 128.9, 127.3, 127.1, 126.4, 109.1, 103.9, 60.6, 55.4, 45.6, 26.1, 22.5, 21.1, 19.9, 14.3

< Example  2> Dihydroxyphenyl series Stereoisomeric Hsp90  Confirmation of binding inhibition

After chiral separation, the binding affinity of each stereoisomer and human recombinant Hsp90? Was confirmed. As a result, it was confirmed that the peak 1 compound showed very good binding affinity (IC ₅₀ = 2.96 nM) to Hsp90α as shown in Table 1, while the compound of peak 2 showed normal binding affinity (IC ₅₀ = 73.6 nM) I could.

< Example  3> Dihydroxyphenyl series Stereoisomeric  Confirming Cancer Cell Proliferation Inhibitory Effect

In order to confirm the dose dependent effect of peak 1 on the growth of cancer cells, H1975 cells were treated with various concentrations of peak 1 for 3 days and cell proliferation level was confirmed.

As a result, as shown in Fig. 2A, the peak 1 compound effectively inhibited the growth of H1975 cells in a dose-dependent manner compared to the gefitinib and peak 2 compounds.

On the other hand, in order to confirm the effect of peak 1 in the Hsp90-dependent signaling process, the expression level of the Hsp90 client protein in H1975 cells was confirmed.

The cell biomarker of Hsp90 inhibition confirmed the differentiation of the client protein and the induction of Hsp70.

As a result, the expression of EGFR, p-EGFR, Her2, Met and Akt was significantly decreased in H1975 treated with Peak 1 as shown in Fig. 2B, whereas increased expression of Hsp70 and Hsp90 was confirmed. The expression level of β-actin, an internal standard, was not changed.

From the above results, it was confirmed that the anti-proliferative effect of the Peak 1 compound in cancer cells was a result of suppression of Hsp90, and Peak 1 was the most effective as compared with the test compound, iressa, peak 2 and geladinamycin (GA) And exhibited excellent anticancer activity.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereby. something to do. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims (9)

A dihydroxyphenyl-based stereoisomer represented by the following formula (1).
[Chemical Formula 1]

In Formula 1,
R ₁ is any one selected from the group consisting of halogen, C ₁ -C 4 alkyl and C 1 -C 4 alkoxy,
R ₂ is any one selected from the group consisting of C 3 -C 6 cycloalkyl, phenyl, halogen, C 1 -C 4 alkyl and C 1 -C 4 alkoxy,
R ₃ is C 1 -C 4 alkyl or C 1 -C 4 alkoxy. The dihydroxyphenyl-based stereoisomer according to claim 1, wherein the stereoisomer is a compound represented by the following formula (2).
(2)

A pharmaceutical composition for preventing or treating a heat shock protein 90 (Hsp90) mediated disease comprising a dihydroxyphenyl stereoisomer represented by the following formula (1) as an active ingredient.
[Chemical Formula 1]

In Formula 1,
R ₁ is any one selected from the group consisting of halogen, C ₁ -C 4 alkyl and C 1 -C 4 alkoxy,
R ₂ is any one selected from the group consisting of C 3 -C 6 cycloalkyl, phenyl, halogen, C 1 -C 4 alkyl and C 1 -C 4 alkoxy,
R ₃ is C 1 -C 4 alkyl or C 1 -C 4 alkoxy. [Claim 3] The pharmaceutical composition for preventing or treating Heat Shock Protein 90 (Hsp90) -mediated disease according to claim 3, wherein the stereoisomer is a compound represented by Formula 2 below.
(2)

[Claim 5] The pharmaceutical composition according to claim 3, wherein the heat shock protein 90 mediated disease is selected from the group consisting of cancer diseases, degenerative neurological diseases and viral infections. The method of claim 5, wherein the cancer is selected from the group consisting of non-small cell lung cancer, breast cancer, ovarian cancer, uterine cancer, pancreatic cancer, lung cancer, gastric cancer, liver cancer, colon cancer, skin cancer, head or neck cancer, brain cancer, larynx cancer, prostate cancer, , Renal cancer, rectal cancer, acute myelogenous leukemia, chronic myelogenous leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia and hematologic malignancies. 6. The method of claim 5, wherein the degenerative neurological disease is selected from the group consisting of stroke, paralysis, memory loss, memory impairment, dementia, forgetfulness, Parkinson's disease, Alzheimer's disease, Pick's disease, Creutzfeld-Kacob disease, Huntington & Wherein the pharmaceutical composition is selected from the group consisting of a pharmaceutical composition for preventing or treating thermal shock protein 90-mediated diseases. A health food for preventing or ameliorating a heat shock protein 90 (Hsp90) mediated disease comprising a dihydroxyphenyl based stereoisomer represented by the following formula (1) as an active ingredient.
[Chemical Formula 1]

In Formula 1,
R ₁ is any one selected from the group consisting of halogen, C ₁ -C 4 alkyl and C 1 -C 4 alkoxy,
R ₂ is any one selected from the group consisting of C 3 -C 6 cycloalkyl, phenyl, halogen, C 1 -C 4 alkyl and C 1 -C 4 alkoxy,
R ₃ is C 1 -C 4 alkyl or C 1 -C 4 alkoxy. [Claim 8] The health food for preventing or improving a disease caused by Heat Shock Protein 90 (Hsp90) according to claim 8, wherein the stereoisomer is a compound represented by the following formula (2).
(2)

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