KR20160096274A - Antifungal composition comprising 2',4'-dihydroxychalcone compound - Google Patents

Abstract

The present invention relates to an antifungal composition comprising a 2,4-dihydroxychalcone compound or a pharmaceutically acceptable salt thereof. In addition, the present invention relates to a pharmaceutical composition for preventing or treating aspergillosis, comprising a 2,4-dihydroxychalcone or a pharmaceutically acceptable salt thereof as an active ingredient. In a patient whose immunity level is degraded, aspergillosis caused by Aspergillus fumigatus is a main cause of infection and death. However, development and use of a pharmaceutical agent capable of treating such infection effectively is very limited up to date. Therefore, the 2,4-dihydroxychalcone compound according to the present invention can be developed and used as an antifungal agent against Aspergillus fumigatus.

Description

An antifungal composition comprising 2 ', 4'-dihydroxychalcone compound as an active ingredient,

The present invention relates to an antifungal composition comprising 2 ', 4'-dihydroxychalcone compound as an active ingredient.

Heat shock protein 90 (Hsp90) is an ATP-dependent molecular chaperone present in all eukaryotes. This plays an essential role in maintaining the proper structure of the client protein. The client proteins for Hsp90 include transmembrane tyrosine kinases (epithelial growth factor receptor and human epidermal growth factor receptor 2), transcription factors (hypoxia-inducible transcription factor 1α and signal transduction and transcription factor 1 (α and signal transducer and activator of transcription 3) and metastable signaling proteins (protein kinase B, IκB kinase and Raf-1) . Hsp90 is involved in various cell functions in normal cells, and is also involved in malignant tumor metastasis and disease progression. Thus, Hsp90 has become a molecular activation target in the treatment of many diseases, including cancer, neurodegenerative diseases and pathogenic infections (viruses, fungi and bacteria).

Invasive fungal pneumonia (aspergillosis) caused by Aspergillus fumigatus in patients with impaired immunity is a major cause of infection and death. Early diagnosis and appropriate treatment can improve clinical outcomes. Aspergillus fumigatus infections can be treated with triazole, polyene or echinocandin drugs, but the number of antifungal drugs available for the treatment of these pathogens is limited. Furthermore, resistance to drugs has occurred and the clinical efficacy of available drugs is declining. Therefore, there is a need for a new therapeutic strategy for life-threatening fungal infections. To improve the efficiency of antifungal drugs and overcome resistance to drugs, Hsp90 in pathogenic fungi is emerging as a potential target for new antifungal agents. In 1994, the first natural product, geldanamycin (GDA), was identified as an Hsp90 inhibitor. Several small-molecule Hsp90 inhibitors have since been developed, including ganetespib and PU3, which show that Hsp90-targeted therapy is evolving.

Korean Patent Laid-Open No. 10-2012-0081690 (published on July 20, 2012)

It is an object of the present invention to provide a composition for an antifungal agent comprising 2 ', 4'-dihydroxychalcone compound or a pharmaceutically acceptable salt thereof as an active ingredient.

In order to achieve the above object, the present invention provides a composition for an antifungal agent comprising 2 ', 4'-dihydroxychalcone compound or a pharmaceutically acceptable salt thereof as an active ingredient .

The present invention also relates to a pharmaceutical composition for preventing or treating fungal pneumonia comprising 2 ', 4'-dihydroxychalcone compound or a pharmaceutically acceptable salt thereof as an active ingredient Lt; / RTI >

The present invention also relates to a pharmaceutical composition for preventing or ameliorating aspergillosis comprising 2 ', 4'-dihydroxychalcone compound or a pharmaceutically acceptable salt thereof as an active ingredient. To provide a food composition.

The present invention relates to a composition for an antifungal agent comprising 2 ', 4'-dihydroxychalcone (SY-032) as an active ingredient. In a patient having decreased immunity, Aspergillus fumigatus Aspergillosis caused by Aspergillus fumigatus is a major cause of infection and death. However, the development and use of drugs that can effectively treat infections are very limited. Therefore, the 2 ', 4'-dihydroxycalcone compound of the present invention can be developed and utilized as an antifungal agent against Aspergillus fumigatus.

FIG. 1 (a) is a schematic view showing the preparation of 3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2H-tetrazolium bromide 5-diphenyl-2H-tetrazolium bromide ; MTT) through analysis, SY-032, and geldanamycin (geldanamycin; Aspergillus fumigatus Fu for GDA) (Aspergillus fumigatus ) Af293. The conidia suspension was diluted in RPMI 1640 containing MTT at a final concentration of 0.1 mg / mL, and plates were incubated for 48 hours. After the culture, the formazan reaction product was extracted and measured. (b) Representative inverted microscope image of Aspergillus fumigatus Af293. Visible differences in mycelial growth with three different GDA and SY-032 concentrations (64, 128 and 256 mg / L) were evident. The scale bar is 200 μm.
Figure 2 is a solid RPMI 1640 media (a) and liquid MMG media (b) from Aspergillus fumigatus Fu's (Aspergillus fumigatus ) SY-032 in Af293 ( In vitro ) antifungal activity. Hyphal growth and pigmentation were deficient after SY-032 treatment. (c) conidiation - results of quantification of specific gene transcripts ( abaA , brlA and wetA ) by real-time PCR. Data were expressed as means ± SD. ** P < 0.01.
Figure 3 (a) shows the results of quantification of transcription of calcineurin ( cnaA ) and transcription factor crzA by real-time PCR. Data were expressed as means ± SD. ** P < 0.01 and * P < 0.05. A variety of SY-032 (0-256 mg / L) combined with a constant concentration of itraconazole (ITC; 0.2 mg / L; b) and caspofungin (CSP; 2.0 mg / In vitro test for concentration vitro) show the effect. The graphs are shown as means ± SD. * P &lt; 0.05. (d) Pu Aspergillus fumigatus (Aspergillus fumigatus ) Af293. Visible differences with respect to three different SY-032 concentrations (32, 64 and 128 mg / L) were apparent in mycelial growth. The scale bar is 50 μm.
Figure 4 (a) shows the Hsp90 domain structure (bottom) of Aspergillus fumigatus as compared to the Hsp90 domain structure (top) of Saccharomyces cerevisiae . The two Hsp90 proteins are highly conserved and consist of the HATPase and HSP90 domains. The pink part is a low complexity part and the green part is a coiled coil part. (b) amino acid sequence comparison of two Hsp90 proteins. The asterisk indicates the amino acids expected to interact with SY-032.
5 shows the results of analysis of SY-032 binding to yeast Hsp90. (a) A comparison docking pose of ligand-13N and SY-032 and yeast Hsp90 (PDB code: 2XX5). (b) Docking model of SY-032 in ATP-binding pocket of yeast Hsp90. 13N and SY-032 are shown in cyan and yellow, respectively. The oxygen, nitrogen and chlorine atoms of 13N and SY-032 were red, blue and green, respectively. The side chains of the ATP-binding site of Hsp90 were colored according to the atomic form (carbon, gray, oxygen, blue; nitrogen, red, sulfur, yellow) and the residue name. Hydrogen bonds are indicated by red dotted lines.

The present inventors have completed a program capable of developing potential Hsp90 inhibitors as a counterpart to pathogenic fungi. The inventors have developed target-focused compound libraries through Hsp90 studies. 2 ', 4'-dihydroxychalcone (SY-032), which exhibits antifungal activity against Aspergillus fumigatus through screening with a target- And completed the present invention.

The present invention provides a composition for an antifungal agent comprising 2 ', 4'-dihydroxychalcone compound or a pharmaceutically acceptable salt thereof as an active ingredient.

Specifically, the compound or a pharmaceutically acceptable salt thereof may inhibit heat shock protein 90 (Hsp90).

Preferably, the fungus may be Aspergillus fumigatus , but is not limited thereto.

The 2 ', 4'-dihydroxychalcone compound of the present invention is represented by the following formula (1) and is also referred to as "SY-032" in the present specification.

[Chemical Formula 1]

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 chalcone compound of the present invention includes not only pharmaceutically acceptable salts, but also all salts, hydrates and solvates which can be prepared by conventional methods.

The present invention also relates to a pharmaceutical composition for preventing or treating fungal pneumonia comprising 2 ', 4'-dihydroxychalcone compound or a pharmaceutically acceptable salt thereof as an active ingredient Lt; / RTI &gt;

The term &quot; aspergillosis &quot; of the present invention is also known as Aspergillosis, and refers to a disease caused by respiratory infections caused by Aspergillus . In an invasive case, the infection spreads systemically, causing tissue damage, thrombus formation due to blood supply disturbance, and even severe sepsis.

The pharmaceutical composition may be in the form of an appropriate carrier, excipient, disintegrant, sweetener, coating agent, swelling agent, lubricant, lubricant, flavoring agent, antioxidant, buffer, bacteriostatic agent, diluent, One or more additives selected from the group consisting of an activator, 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 a subject in a conventional manner via the intradermal route.

The preferred dosage of the compound may vary depending on the condition and body weight of the subject, the type and degree of disease, the drug form, the administration route and the period, 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.

The present invention also relates to a pharmaceutical composition for preventing or ameliorating aspergillosis comprising 2 ', 4'-dihydroxychalcone compound or a pharmaceutically acceptable salt thereof as an active ingredient. To provide a food composition.

In one 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, sugars, ice cream products, dairy products, meat products, fish meat products, tofu or glue, edible oils, noodles, Dried products, cut products, fruit juice, vegetable juice, mixed juice, chips, noodles, livestock processed foods, processed fish products, dairy products such as ice, ginseng products, kimchi pickles, It can be used for the production of foods such as foods, fermented milk foods, bean curd foods, cereal foods, fermented microorganism foods, confectionery bakery, condiments, meat products, acidic drinks, licorice products and herbal products.

Hereinafter, the present invention will be described in detail with reference to examples which do not limit the present invention. It should be understood that the following embodiments of the present invention are only for embodying the present invention and do not limit or limit the scope of the present invention. It is therefore to be understood that within the scope of the appended claims, the invention may be practiced otherwise than as specifically described herein.

< Experimental Example  >

The following experimental examples are intended to provide experimental examples that are commonly applied to the respective embodiments according to the present invention.

1. Strain and compound

Antifungal drugs are wild-type Aspergillus fumigatus Fu (Aspergillus fumigatus ) Af293 (Fungal Genetics Stock Center, Kansas City, MO, USA). Caspofungin (CSP), itraconazole (ITC), and geldanamycin (GDA) were purchased from Sigma Chemical Co. (St. Louis, Mo., USA).

2. 2 ', 4'-dihydroxycalcone (2', 4'- 다 히시 시크콜 ) Synthesis of

2 ', 4'-dihydroxychalcone (SY-032) was synthesized as follows. A mixture of compound 3 (0.30 g, 1.29 mmol), benzaldehyde (0.14 mL, 1.42 mmol) and KOH (0.6 g) dissolved in 12 mL methanol was mixed at room temperature for 4 days. The mixture was neutralized to pH 6 with 1 N HCl, then extracted with ethyl acetate. The organic layer was washed three times with saturated sodium bicarbonate solution, dried over sodium sulfate, concentrated under reduced pressure, and purified by medium pressure chromatography to yield compound 2 in 73% yield. liquid chromatography (MPLC, Biotage SNAP HP-Sil column). Rf = 0.28 (1: 9 ethyl acetate: hexane). ^{1 H NMR (400 MHz, CDCl} 3) δ 7.77 (d, J = 8.8 Hz, 1H), 7.69 (d, J = 15.6 Hz, 1H), 7.61 (d, J = 16.0 Hz, 1H), 7.56-7.54 (m, 2H), 7.34-7.31 ( m, 3H), 6.53 (dd, J = 8.6 Hz, 2.0 Hz, 1H), 6.47 (d, J = 2.0 Hz, 1H), 6.06-5.96 (m, 2H) , 5.44-5.37 (m, 2H), 5.29-5.22 (m, 2H), 4.55-4.51 (m, 4H). ¹³ C NMR (100 MHz, CDCl ₃ ) δ 189.61, 162.78, 159.03, 141.09, 135.01, 132.55, 132.23, 132.02, 129.56, 128.43, 127.91, 126.95, 122.02, 117.71, 117.59, 106.06, 99.81, 68.89, 68.55. ESI MS ( m / e ) = 321 [M + 1] &lt; ⁺ &gt;. Compound 2 was prepared by reacting 13 mg of bis (triphenylphosphine) palladium (II) dichloride (13 mg) dissolved in tetrahydrofuran (4 mL) and ammonium formate ) (80 mg) was added and mixed for 30 minutes at 120 ° C under microwave irradiation (Biotage Initiator). The reaction mixture was diluted with ethyl acetate. The organic layer was washed with water, dried over sodium sulfate, concentrated under reduced pressure and purified by MPLC to give compound 1 in 39% yield. Rf = 0.24 (1: 4 ethyl acetate: hexane ^{1 H NMR (400 MHz, CDCl} 3) δ 13.41 (s, 1H), 7.88 (d, J = 15.6 Hz, 1H), 7.84 (d, J = 9.2 Hz , 1H), 7.66-7.63 (m, 2H), 7.57 (d, J = 15.2 Hz, 1H), 7.44-7.42 (m, 3H), 6.47 (d, J = 2.4 Hz, 1H), 6.45 (s, ESI MS ( m / e ) = 241 [M + 1] ⁺ (scheme 1).

[Reaction Scheme 1]

3. Sensitivity Analysis

The antifungal susceptibility assay was performed using the previously reported 3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2H-tetrazolium bromide (3- 2,5-diphenyl-2H-tetrazolium bromide (MTT) method. RPMI 1640 medium containing L-glutamine and without sodium bicarbonate was buffered with 165 mM morpholinepropanesulfonic acid buffer (pH 7.0) and used as a test medium Respectively. All test compounds were dissolved in 1.28 g / L dimethyl sulfoxide. The compounds were serially diluted to obtain final drug concentrations ranging from 0.06 to 256 mg / L. The inoculum suspension was prepared according to the hemocytometer and diluted with RPMI 1640 containing 0.2 mg MTT / mL to obtain an inoculum in the range of 1 × 10 ⁴ to 5 × 10 ⁴ conidia / mL. The same volume (100 μL) of inoculum and compound was then added to the microplate, so that the final MTT concentration was 0.1 mg / mL. The plate was incubated at 37 DEG C for 48 hours. After incubation, the formazan analytical product was extracted and measured at an optical density of 540 nm. Experiments were performed independently 3 times, each 4 times.

4. Real time Reverse transcription ( Real - time reverse transcription ) - PCR ( RT - PCR )

In order to investigate the association between antifungal agents and conidiation , we used brlA , abaA and wetA Gene expression was measured. In addition, in order to measure the test drug effect on the calcineurin pathway, cnaA And crzA were analyzed. Minimal suspension (5 × 10 ⁵ conidia / mL) was inoculated into glucose minimal medium (MMG) and cultured at 37 ° C. for 48 hours. RNA extraction, cDNA synthesis and RT-PCR were performed as previously reported. The expression ratio was normalized to elongation factor 1 [alpha] (EF1 [alpha]) and calculated according to the [Delta] Ct method. All experiments were performed in triplicate and data were expressed as mean ± standard deviation (SD).

4. Microscopic analysis

Microscopic photographs were obtained using an Olympus Inverted Microscope IX50 equipped with a Lumenera Infinity camera (Olympus Corporation, Tokyo, Japan).

5. Statistical Analysis

Asymmetric Student's t- test was used for two separate sets of comparisons for independent samples. A P value of less than 0.05 was considered statistically significant. The MTT conversion of the treated and control groups was post hoc Tukey comparison using ANOVA. When the P value is lower than 0.05, the deviation is considered important. Statistical analysis was performed with IBM SPSS statistics 21.0 (IBM, Armonk, New York, USA).

6. Domain structure analysis and docking ( docking ) Research

The domain structure of Hsp90 was analyzed using the domain analysis site SMART (http://smart.embl-heidelberg.de), protein alignment was performed using EMBOSS needle (version 6.6.0) (http://www.ebi.ac. uk / Tools / psa / emboss_needle /). SY-032 and yeast Hsp90: in silico docking with (PDB code 2XX5) (In silico docking) was completed using the AutoDock4.2 program downloaded from the Scripps Research Institute at the Molecular Graphics Laboratory. AutoDock4.2 program has been selected because it uses a genetic algorithm to create the poses (poses) of an internal ligand binding site known or predicted to take advantage of the Lamarckian version of the genetic algorithm, in situ The intra-structural changes applied by the molecules after optimization were used as the next poses to the result. For docking experiments, Gasteiger charges were verified on the X-ray structure of the HSP90 domain N-terminal with SY-032 using tools from the AutoDock suite. A grid box midway on the Hsp90 domain N-terminus, defined as 50_50_50 points and 0.375A spacing, was selected for ligand docking experiments. The root-mean-square tolerance for grouping is cut off to 1 Å in each docking reaction, while the docking parameters consist of set population size 150, number of households 27,000 and number of evaluations 25,000,000, The water reaction is set to 100. Docking models of yeast Hsp90 and SY-032 were drawn, and the rendering of the model was made using PyMol (DeLano Scientific).

< Example  1> Aspergillus Pumigatous ( Aspergillus fumigatus )on  About SY -032 antifungal activity

The antifungal activity of SY-032 was tested and the minimum inhibitory concentration (MIC) was determined. As shown in Figure 1A, SY-032 was more effective against Aspergillus fumigatus than GDA, a well known Hsp90 inhibitor. Compared with GDA, a significant decrease in MTT conversion was observed at SY-032 concentration ≥ 64 mg / L. The MIC ₅₀ of SY-032 (MIC inhibiting 50% of growth) was between 64 mg / L and 128 mg / L. The MIC ₅₀ of GDA was higher than 256 mg / L (Fig. 1A). In addition, the inhibitory effect of SY-032 was tested using a reversed-phase microscope. Visible differences in mycelial densities and growth rates between SY-032 and GDA at 64, 128 and 256 mg / L were clearly observed (FIG. 1B). Treatment with SY-032 (256 mg / L) severely reduced mycelial growth.

< Example  2> nutrition growth ( vegetative growth ) And titration Minster  formation( 공지 ) SY -032

By treatment with test compounds, Radial growth of Aspergillus fumigatus was inhibited in RPMI 1640 agar medium. SY-032 (8 mg / L) reduced relative radial growth by 20% when compared to the control. Also, the treated colonies were deficient in green pigmentation, indicating little formation of conidia (Fig. 2a). This phenomenon was also clearly observed in liquid culture. As shown in Figure 2b, green pigmentation was completely deficient after incubation with SY-032. Because of the strong suppression of myelogenesis by drug treatment, the present inventors have found that abaA , brlA, and wetA , which encode transcription factors that regulate asexual development in Aspergillus species Expression was quantitated. Compared with the control group, expression of all three genes was significantly reduced (about 3 to 5 times) by treatment with SY-032 (Fig. 2C).

< Example  3> SY Suppressed by -032 Calcineurin ( calcineurin ) Route

The calcineurin signaling pathway is required for proper mycelial growth and is activated in response to cell wall stress in Aspergillus fumigatus . To investigate the effect of SY-032 on calcineurin signaling, we used two major signaling components, cnaA And crzA were quantified. cnaA Expression was reduced by about 1.5-fold by SY-032, while the zinc finger transcription factor crzA (downstream of cnaA ) significantly decreased under SY-032 (Fig. 3a). The degree of inhibition was expressed as cnaA The crzA was larger. In yeast, CrzA has been reported to play a partial role in azole and echinocandin resistance. We tested the SY-032 effect in combination with ITC (azole, 0.2 mg / L) and CSP (echinocandin, 2.0 mg / L). ITC alone showed minimal inhibitory activity. However, in combination with ITC and SY-032 (greater than 64 mg / L), the inhibitory activity was significantly improved (Figure 3b). Furthermore, the combination of CSP and increased concentration of SY-032 continuously inhibited Aspergillus fumigatus and completely inhibited growth at a concentration of 256 mg / L (Fig. 3C). In addition, the inhibitory effect of antifungal agents ITC and CSP in combination with SY-032 was tested using an optical microscope. Mycelial growth was significantly reduced by the combination treatment. Surprisingly, when SY-032 was also used in combination with ITC, the germination rate also decreased (FIG. 3D).

< Example  4> Hsp90 of ATPase  Combined with domain SY -032

Aspergillus fumigatus Hsp90 (AFUA_5G04170) consists of 706 amino acid residues and includes the C-terminal HATPase c domain (AA 28-177) and the HSP90 domain (AA 183-706). Aspergillus fumigatus Fu (Aspergillus fumigatus) and Celebi as Saccharomyces My jiae access (Saccharomyces The Hsp90 protein derived from S. cerevisiae is highly conserved (Fig. 4A), exhibiting 75.3% identity and 85.8% similarity, and the amino acids expected to interact with SY-O32 are completely consistent (Fig. 4B). To represent the combined mode with the SY-032 in the ATP- binding site of Hsp90, Celebi as Saccharomyces My jiae access (Saccharomyces Docking studies were performed using the Hsp90 crystal structure (PDB code: 2XX5) of S. cerevisiae , its unmodified ligands 13N and SY-032. Using the AutoDock4.2 program, SY-032 was docked with the 3D coordinates of the Hsp90 N terminal domain. SY-032 and 13N in the ATP binding site of Hsp90 showed that SY-032 binds to Hsp90 in a manner similar to Hsp90 unmodified ligand 13N (FIG. 5). The resorcinol ring and the phenyl ring of SY-032 were overlapped with a 13 N resorcinol ring and phenyl ring in the ATP-binding site of Hsp90, the enone site of SY-032 being 13 N Macrolactam was applied to other structures derived from the ring. Specifically, the two hydroxyl groups and the carbonyl oxygen atom of the resorcinol ring of SY-032 form hydrogen bonds with Asp79, Asn37 and conserved water molecules in the hydrophilic region of the pocket. On the other hand, the phenyl ring of SY-032 was applied to hydrophobic sites formed by the amino acid residues Met84, Leu89, Phe124, Val136 and Trp148 and closed lipophilic contacts with residues Met84, Leu89, Phe124, Val136 and Trp148, (Fig. 5). Taken together, the binding of SY-032 to yeast Hsp90 contributed to hydrogen-bonding and hydrophobic interactions with the resorcinol ring, carbonyl oxygen atom and phenyl ring of SY-032, using the Lamarckian genetic algorithm (Gb) and inhibition constants (Ki) were -7.65 kcal / mol and 2.46 [mu] M, respectively.

Claims (5)

2 ', 4'-dihydroxychalcone compound or a pharmaceutically acceptable salt thereof as an active ingredient. The antifungal composition according to claim 1, wherein the compound or a pharmaceutically acceptable salt thereof binds to heat shock protein 90 (Hsp90) to inhibit it. The composition for an antifungal agent according to claim 1 or 2, wherein the fungus is Aspergillus fumigatus . A pharmaceutical composition for preventing or treating fungal aspergillosis comprising 2 ', 4'-dihydroxychalcone compound or a pharmaceutically acceptable salt thereof as an active ingredient. A health food composition for preventing or ameliorating aspergillosis comprising 2 ', 4'-dihydroxychalcone or a pharmaceutically acceptable salt thereof as an active ingredient.

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