KR20130044838A - Anticancer composition containig the benzochalcone - Google Patents

Abstract

PURPOSE: A benzochalcone compound is provided to restrain growth restraining and apoptosis of cancer cells by activating caspase-7 enzyme activation which is a protein controlling apoptosis. CONSTITUTION: A manufacturing method of a benzochalcone compound comprises a step of dissolving 2-hydroxy-1-acetonaphthone and 2,4,6-trimethoxybenzaldehyde into ethanol; a step of adding 50% KOH aqueous solution to the solution mixture; a step of stirring and cooling the solution mixture; a step of neutralizing the solution mixture; a step of vacuum filtrating and washing a benzochalcone solid compound generated during the neutralization; a step of recrystallizing the washed benzochalcone compound in the ethanol.

Description

Anticancer agent composition containing benzochalcone as an active ingredient {ANTICANCER COMPOSITION CONTAINIG THE BENZOCHALCONE}

The present invention relates to a pharmaceutical composition for the prevention and treatment of cancer diseases, and more particularly, it is represented by the following Chemical Formula 1 having growth inhibition and killing activity of cancer cells by caspase-7 enzyme activity, an apoptosis regulating protein. New benzochalcone compounds (E-1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one, ( E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one) and pharmaceutical compositions for the prevention and treatment of cancer diseases including the same It is about.

[Formula 1]

The name caspase comes from the cystein aspartate protease and is a protein with an executive character of apoptosis. Caspase-7 is one of apoptosis related cystein peptidases found in most mammals and humans. Caspase-7 is present as an inactive proprotein, which is then degraded by caspase-9. When a signal related to apoptosis is provided, caspase-7 is activated to induce apoptosis. Therefore, since caspase-7 cleavage is closely related to induction of apoptosis, the caspase-7 cleavage effect is recognized as a signal for apoptosis of cancer cells affected by apoptosis. It can be said that it has utility as.

Flavonoids have a structural skeleton of C 6 -C 3 -C 6, which is called a benzoflavonoid when the benzene ring is attached to another C 6 skeleton to have a naphthalene group. Chalcone is also a flavonoid, so if one benzene ring is changed to a naphthalene group, it becomes benzochalcone. Since the hydrophobicity is increased in the case of benzochalcon than the known chalcone, permeation of the cell membrane becomes a little easier, and thus a change in biological activity can be expected. Therefore, the present inventors synthesized a compound having a new structure by adding three methoxy groups and one hydroxy group having a skeleton of benzochalcone and named it DK78. Since the DK78 compound, a kind of benzochalcone, has been reported to have a cleavage effect of caspase-7 as a new compound that has not been reported so far, further experiments related to cell cycle inhibition were performed to confirm whether the compound has an efficacy as an anticancer agent. The invention was completed.

Conventional techniques using a chalcone or a derivative thereof as a pharmaceutical composition have been disclosed in Korean Patent No. 10-0567125 (A matrix composition for inhibiting matrix metalloprotease activity containing a chalcone or a derivative thereof) by inhibiting MMP activity. Patent No. 10-0751899 (new chalcone derivatives, pharmaceutically acceptable salts thereof, preparation method thereof and glycosidase containing them as an active ingredient) which has the effect of preventing and treating diseases caused by glycosidase. Composition for the prevention and treatment of diseases caused by).

It is an object of the present invention to provide a novel benzochalcone compound (E-1- (2-hydroxynaphthalene-) having the inhibitory effect on the growth and killing of cancer cells by the caspase-7 enzyme activity, which is an apoptosis regulating protein. 1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one, (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2 , 3,4-trimethoxyphenyl) prop-2-en-1-one) and a pharmaceutical composition for preventing and treating cancer diseases including the same.

Further, another object of the present invention is to provide a novel benzochalcone compound (E-1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2 -En-1-one, (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one) To provide a dietary supplement for prevention and improvement.

In order to achieve the above object, the present invention is a benzochalcone compound represented by the following formula (E-1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimeth Methoxyphenyl) prop-2-en-1-one, (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one ) Or a pharmaceutically acceptable salt thereof.

[Formula 1]

In addition, the present invention is a benzochalcone compound represented by the formula (1) (E-1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop 2-en-1-one, (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one) (1) 2-hydroxy-1-acetonaphthone and 2,4,6-trimethoxybenzaldehyde (2,4,6-trimethoxybens-aldehyde) are dissolved in ethanol. (2) adding a 50% KOH aqueous solution to the mixed solution according to step (1); and (3) cooling the mixed solution prepared by step (2) at room temperature and then cooling it. And (4) neutralizing by adding 6 N HCl solution to the mixed solution cooled by the step (3); and (5) subjecting the benzochalcon compound solid produced in the neutralization process of the step (4) to filtration. Washing with ethanol; And (6) re-refining the benzochalcon compound solid washed by the step (5) in ethanol; a benzochalcone compound (E-1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one, (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4 -trimethoxyphenyl) prop-2-en-1-one) is provided.

In addition, the present invention is a caspase is apoptosis control protein for cancer cells selected from the group consisting of colon cancer, stomach cancer, prostate cancer, breast cancer, kidney cancer, liver cancer, brain tumor, lung cancer, uterine cancer, colon cancer, bladder cancer, pancreatic cancer, blood cancer (benzospacone) compound (E-1- (2-hydroxynaphthalen-1-yl) -3- (1) having a growth inhibitory and killing effect of cancer cells by enzyme activity (caspase) -7 2,3,4-trimethoxyphenyl) prop-2-en-1-one, (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop Provided is a pharmaceutical composition for the prevention and treatment of cancer diseases, including 2-en-1-one) and pharmaceutically acceptable salts.

The pharmaceutical composition is characterized in that it comprises one or more pharmaceutically acceptable carriers, diluents or excipients.

The pharmaceutical composition is characterized in that it comprises a second anticancer agent or anticancer adjuvant.

The second anticancer agent is interferon, interleukin-2, paclitaxel, vincristine, vinblastin, doxorrubicin, etoposide, etoposide, Irinotecan hydrochloride, cisplatin, amsacrine, cytosine arabinoside, fluorouracil and taxol It is done.

In addition, the present invention is a benzochalcone compound represented by the formula (1) (E-1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop 2-en-1-one, (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one) and foodologically Provided are nutraceuticals for the prevention and improvement of cancer diseases, including acceptable food supplement additives.

The health functional food is characterized in that the tablet, capsule, pills or liquid form.

According to the present invention as described above, a novel benzochalcone compound (E-1- (2-hydroxynaphthalene) having growth inhibition and killing activity of cancer cells by caspase-7 activity, which is an apoptosis regulating protein -1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one, (E) -1- (2-hydroxynaphthalen-1-yl) -3- ( 2,3,4-trimethoxyphenyl) prop-2-en-1-one) and a pharmaceutical composition for the prevention and treatment of cancer diseases and the health functional food for the prevention and improvement of cancer diseases, thereby preventing cancer diseases And not only can be usefully used to treat, there is an effect that can be usefully used in health food for the prevention and improvement of cancer diseases.

1 is a hydrogen nuclear magnetic resonance spectra of the present invention benzochalcone compound (DK78). (400MHz Bruker nuclear magnetic resonance spectrometer)
2 is a carbon nuclear magnetic resonance spectroscopy spectrum of the present invention benzochalcone compound (DK78). (Using 100MHz Bruker nuclear magnetic resonance spectrometer)
Figure 3 shows the caspase activity effect of the present invention benzochalcone compound (DK78) in HCT116 cells by Western blot method.
Figure 4 shows the ability of the benzochalcon compound (DK78) of the present invention inhibit HCT116 colon cancer cell proliferation.
Figure 5 is analyzed by the flow cytometer apoptosis effect of the present invention benzochalcon compound (DK78) in HCT116 cells (2N, dploid DNA; 4N, tetraploid DNA. M1, sub-G1; M2, G1; M3, S; M4, G2 / M)

Hereinafter, the present invention will be described in detail.

The present invention is a benzochalcone compound represented by the following formula (E-1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2 -En-1-one, (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one) or a pharmaceutically acceptable thereof To provide salts.

[Formula 1]

The present invention is a benzochalcone compound represented by the formula (1) (E-1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2 -En-1-one, (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one) (1) dissolving 2-hydroxy-1-acetonaphthone and 2,4,6-trimethoxybenzaldehyde (2,4,6-trimethoxybens-aldehyde) in ethanol And (2) adding 50% KOH aqueous solution to the mixed solution according to step (1); and (3) cooling the mixed solution prepared by step (2) at room temperature and then cooling; (4) neutralizing by adding 6 N HCl solution to the mixed solution cooled by step (3); and (5) subjecting the benzocalcon compound solid produced in the neutralization process of step (4) to sub-filtration, Washing with ethanol; And (6) re-refining the benzochalcon compound solid washed by the step (5) in ethanol; a benzochalcone compound (E-1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one, (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4 -trimethoxyphenyl) prop-2-en-1-one) is provided.

The present invention is a caspase (caspase) which is an apoptosis regulating protein for cancer cells selected from the group consisting of colon cancer, stomach cancer, prostate cancer, breast cancer, kidney cancer, liver cancer, brain tumor, lung cancer, uterine cancer, colon cancer, bladder cancer, pancreatic cancer, and blood cancer. Benzochalcone compound represented by the formula (1) having growth inhibition and killing activity of cancer cells by enzyme activity) (E-1- (2-hydroxynaphthalen-1-yl) -3- (2, 3,4-trimethoxyphenyl) prop-2-en-1-one, (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2 -en-1-one) and a pharmaceutically acceptable salt comprising a pharmaceutical composition for the prevention and treatment of cancer diseases.

The pharmaceutical composition may comprise one or more pharmaceutically acceptable carriers, diluents or excipients. The carrier, diluent or excipient may 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 and mineral oil.

The pharmaceutical composition may include a second anticancer agent or an anticancer adjuvant. The second anticancer agent or anticancer agent includes interferon, interleukin-2, paclitaxel, vincristine, vinblastin, doxorrubicin, etoposide (etoposide), irinotecan hydrochloride, cisplatin, amsacrine, cytosine arabinoside, fluoro uracil and taxol It is not limited.

Benzochalcone compound (E-1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one of the present invention , Pharmaceutical compositions containing (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one), respectively, according to conventional methods. It may be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like oral formulations, external preparations, suppositories or sterile injectable solutions. Specifically, when formulated, it may be prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, and the like which are commonly used. Solid preparations for oral administration of the benzochalcone compound (E-1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2- At least one excipient in en-1-one, (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one) For example, it may be prepared by mixing starch, calcium carbonate, sucrose, lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral use include suspensions, solutions, 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 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. As the base of the suppository, utopsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Benzochalcone compound (E-1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one of the present invention Preferred dosages of (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one) may be determined by the patient's condition, body weight and disease. Depending on the degree, drug form, route of administration, and duration, it may be appropriately selected by those skilled in the art. However, for the desired effect, the benzochalcone compound (E-1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2 of the present invention -En-1-one, (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one) or a pharmaceutically acceptable thereof Possible salts may be administered in 0.0001 to 100 mg / kg, preferably in an amount of 0.001 to 100 mg / kg divided once to several times daily. Benzochalcone compounds (E-1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-ene of the present invention in the entire pharmaceutical composition -1-one, (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one) or a pharmaceutically acceptable salt thereof Should be included in 0.0001 to 10% by weight, preferably 0.001 to 1% by weight.

The pharmaceutical composition of the present invention can be administered to mammals such as stomach, mouse, livestock, human, etc. by various routes, and the mode of administration can be administered by injection in various ways such as oral, rectal, intravenous, intramuscular, subcutaneous. .

In addition, the present invention is a benzochalcone compound represented by the formula (1) (E-1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop 2-en-1-one, (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one) and foodologically Provided are nutraceuticals for the prevention and improvement of cancer diseases, including acceptable food supplement additives. For example, dietary supplements include meat, sausages, breads, chocolates, candy, snacks, confectionery, pizzas, ramen noodle, dairy products including gums, ice creams, various soups, beverages, teas, drinks, alcoholic beverages, It can be prepared by adding a benzohydroxymethoxychalcone compound or a pharmaceutically acceptable salt thereof to a vitamin complex and the like.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not construed as being limited by these embodiments.

Example 1 Synthesis of Benzochalcone Compound (DK78)

In the present invention, the benzochalcone compound represented by Chemical Formula 1 (DK78) (E-1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2 -En-1-one, (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one) is shown in Scheme 1 below. The method was synthesized as follows.

[Reaction Scheme 1]

2-hydroxy-1-acetonaphtone ( I , 1.86 g, 10 mmol) and 2,4,6-trimethoxybenzaldehyde ( II , 1.96 g, 10 mmol) were dissolved in 50 ml ethanol while stirring and the temperature was decreased. After about 3 ^° C., 3 ml of 50% KOH solution was slowly added. After stirring for 48 hours at room temperature, the mixture was cooled with ice water, neutralized with 6N hydrochloric acid (HCL), and the solid formed during the neutralization was filtered under reduced pressure and washed with cold ethanol (DK-78) (E). -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one, (E) -1- (2-hydroxynaphthalen -1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one) was obtained. This benzochalcon compound was recrystallized in ethanol to give pure benzochalcon compound (DK-78) in a yield of 79% (2.87 g). Mp; 91-93 ^o C.

In order to confirm the production of the benzochalcon compound (DK-78) was confirmed through the hydrogen nuclear magnetic resonance spectra (Bruker 400MHz) and carbon nuclear magnetic resonance spectrum (Bruker 100MHz). The device used was a Bruker 400 MHz device. Hydrogen nuclear magnetic resonance spectra and carbon nuclear magnetic resonance spectra are shown in FIGS. 1 and 2, respectively, and chemical mobility is as follows.

1 H-NMR (400 MHz) δ (multiple, filling constant / Hz): 2.80 (dd, 16.5, 3.0), 3.43 (dd, 16.5, 13.8), 3.79 (s), 3.84 (s), 3.86 (s) , 5.87 (dd, 13.8, 3.0), 6.91 (d, 8.8), 7.25 (d, 9.0), 7.32 (d, 8.8), 7.49 (ddd, 8.0, 6.9, 1.1), 7.68 (ddd, 8.5, 6.9, 1.1), 7.93 (d, 8.0), 8.15 (d, 9.0), 9.39 (d, 8.5)

13C-NMR (100 MHz) δ: 43.7, 56.0, 60.4, 61.4, 74.2, 108.0, 111.7, 119.0, 122.1, 123.8, 124.7, 125.0, 128.7, 128.8, 129.5, 130.9, 137.6, 141.7, 151.3, 154.0, 163.5, 193.2

The benzochalcone compound (DK78) (E-1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one, ( E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one) is a new substance that has not been reported to date and has a molecular formula of C22H20O5. , Molecular weight is 364.1.

Example 2. Caspase-7 cleavage effect of benzochalcone compound (DK78)

Caspase-9 and caspase-7 enzymes, which induce apoptosis, are normally inactive precursors, and are cleaved and activated when apoptosis stimulus signals are transmitted. In the present invention, in order to confirm the anticancer effect of the benzochalcone compound (DK78), the caspase-7 and caspase-9 activating the cleaved caspase-7 and caspase-9 of the active form using Western blot analysis The degree of fragmentation of the substrate protein poly (ADP-ribose) polymerase (PARP) was investigated.

Invitrogen Life from HCT116 cells (human colorectal cancer cell line) from the American Type Culture Collection (ATCC) and containing 10% Fetal Bovine Serum (Invitrogen Life Technologies) and Antibiotic-Antimycotic solution (Invitrogen Life Technologies) Technologies). Cultures were added to a 60-mm cell culture dish every 2 days and incubated in a 37 ° C., 5% CO ₂ incubator while maintaining a seed density of 1 × 10 ⁶ . The cultured cells were treated with a 20 μM concentration of benzochalcone compound (DK78), and cells were harvested after 0, 6, 12, and 24 hours. Harvested cells for 30 minutes by adding 20 mM HEPES (pH 7.2), 1% Triton X-100, 10% glycerol, 150 mM NaCl, 10 μg / ml leupeptin, 1 mM PMSF After lysing the cells, the cells were harvested by high-speed centrifugation, and the cell lysate was harvested. The protein lysate prepared to contain the same protein was subjected to SDS-polyacrylamide gel electrophoresis. Proteins present in the isolate were separated. Electrophoretically separated proteins were transferred to a polystyrene membrane, followed by cleavage caspase-7. Primary antibodies that bind to caspase-9 and PARP proteins (purchased from Cell Signaling Technology, Inc.) and primary antibodies that recognize glyceraldehyde-3-phosphate dehydrogenase (GAPDH) that do not alter protein expression as controls (from Santa Cruz technology, Inc.) 5 hours each, and then a secondary antibody (purchased by Cell Signaling Technology, Inc.) that prints the primary antibody was reacted for 1 hour. The change in expression of each protein on the X-ray film was analyzed using a Chemiluminescence detection system (Amersham Pharmacia Biotech, Piscataway, NJ).

As a result, as shown in FIG. 3, when the benzochalcone compound (DK78) was treated to HCT116 colon cancer cells, the amount of the control protein GAPDH did not change, but the cleaved caspase-9 and caspase-7 proteins were increased after 24 hours of treatment. I confirmed the fact. In addition, PARP protein is a substrate protein cleaved by caspase-7 enzyme activity, it was observed that PARP protein also produced 89-kDa size fragments 24 hours after the benzochalcon compound (DK78) treatment. These results indicate that the benzochalcone compound (DK78) of the present invention induces caspase-9 and caspase-7 enzyme activities.

Example 3 Growth Inhibition of Colorectal Cancer Cell Line by Benzocalcon Compound (DK78)

Growth inhibition of HCT116 colon cancer cells by the benzochalcone compound (DK49) of the present invention was measured using a CCK-8 kit (Cell Counting Kit-8; Dojindo, Japan). Dispense 1 x 10 ⁴ HCT116 colorectal cancer cells into a 96-well cell culture plate and treat benzochalcone compounds (DK78) at 0, 5, 10, and 20 μM concentrations, and after 24 and 48 hours, respectively, use CCK-8 solution 10 Μl was added. After 2 hours, the cell viability was measured at 450 nm to analyze cell viability.

As a result, as shown in FIG. 4, the cell growth ability increased with time in the control cell group which had not been treated, but the cell growth was time-dependently in the cell group treated with benzocalcon compound (DK78). This has been reduced. These results indicate that apoptosis of HCT116 colon cancer cells is induced by the benzochalcone compound (DK78), thereby inhibiting cancer cell proliferation.

Example 4 Induction of Apoptosis of Colon Cancer Cell Lines by Benzocalcon Compound (DK78)

The main process of apoptosis by drug treatment can be confirmed from the contraction of the cells and the accompanying chromatin condensation and DNA fragmentation of DNA. This phenomenon occurs when the DNA content is measured in the cell cycle (Cell cycle) is less than the 2N DNA content of G1 phase cells (sub-G1) appears. Therefore, cells having a sub-G1 DNA content are generally considered to be cells in which apoptosis is in progress. In the present invention, in order to determine the effect of induction of apoptosis by the benzochalcone compound (DK78) in colorectal cancer cells was measured intracellular DNA content using a flow cytometer (BD Science, USA).

HCT116 cells were treated with benzochalcone compound (DK78) and cells were harvested 24 hours later. The cells were fixed by adding 70% ethanol, and the DNA was stained by reacting PI (Propidium Iodine) for 30 minutes, and then the amount of sub-G1 DNA was analyzed by flow cytometry.

As a result, as shown in Fig. 5, when the benzochalcone compound (DK78) of the present invention was treated at a concentration of 20 μM, the sub-G1 DNA content was increased from 4.73% of control cells to 10.09% and 69.44%, respectively, in a time-dependent manner. . Therefore, it was confirmed that the benzochalcon compound (DK78) according to the present invention induces apoptosis of HCT116 colon cancer cells.

Having described specific portions of the present invention in detail, it will be apparent to those skilled in the art that this specific description is only a preferred embodiment and that the scope of the present invention is not limited thereby. It will be obvious. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (8)

Benzochalcone compound represented by the following formula (1) (E-1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-ene- 1-one, (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one) or a pharmaceutically acceptable salt thereof.
[Formula 1]

A benzochalcone compound represented by formula 1 of claim 1 (E-1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2- In the method for producing en-1-one, (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one),
(1) dissolving 2-hydroxy-1-acetonaphthone and 2,4,6-trimethoxybenzaldehyde (2,4,6-trimethoxybens-aldehyde) in ethanol ;
(2) adding a 50% KOH aqueous solution to the mixed solution of step (1);
(3) stirring the mixed solution prepared by step (2) at room temperature and then cooling;
(4) neutralizing by adding 6N HCl solution to the mixed solution cooled by step (3);
(5) sub-filtration of the benzochalcon compound solid produced in the neutralization process of step (4) and washing with ethanol; And
(6) benzochalcone compound (E-1- (2-hydroxynaphthalen-1-yl)-, comprising the step of re-refining the benzochalcon compound solid washed in step (5) in ethanol; 3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one, (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4- trimethoxyphenyl) prop-2-en-1-one).
Caspase-7, an apoptosis regulating protein for cancer cells selected from the group consisting of colon cancer, stomach cancer, prostate cancer, breast cancer, kidney cancer, liver cancer, brain tumors, lung cancer, uterine cancer, colon cancer, bladder cancer, pancreatic cancer and blood cancer A benzochalcone compound represented by Formula 1 of claim 1 having growth inhibitory and killing effect of cancer cells (E-1- (2-hydroxynaphthalen-1-yl) -3- (2,3) , 4-trimethoxyphenyl) prop-2-en-1-one, (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2- en-1-one) and a pharmaceutically acceptable salt comprising a pharmaceutical composition for the prevention and treatment of cancer diseases.
The method of claim 3, wherein
The pharmaceutical composition is a pharmaceutical composition for the prevention and treatment of cancer diseases, characterized in that it comprises one or more pharmaceutically acceptable carriers, diluents or excipients.
The method of claim 3, wherein
The pharmaceutical composition is a pharmaceutical composition for preventing and treating cancer diseases, characterized in that it comprises a second anticancer agent or an anticancer adjuvant.
The method of claim 5, wherein
The second anticancer agent is interferon, interleukin-2, paclitaxel, vincristine, vinblastin, doxorrubicin, etoposide, etoposide, Irinotecan hydrochloride, cisplatin, amsacrine, cytosine arabinoside, fluorouracil and taxol A pharmaceutical composition for preventing and treating cancer diseases.
A benzochalcone compound represented by the formula (1) of claim 1 (E-1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2- En-1-one, (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2,3,4-trimethoxyphenyl) prop-2-en-1-one) and food acceptable foods Health functional foods for the prevention and improvement of cancer diseases, including supplementary additives.
The method of claim 7, wherein
The health functional food is a health functional food for cancer disease prevention and improvement, characterized in that the tablet, capsule, pills or liquid form.

Images