KR20140052371A - A dihydropyrazolecarbothioamide derivative, method of preparing the same, and anti-cancer agent comprising the same - Google Patents

Abstract

The present invention relates to a dihydropyrazolecarbothioamide derivative, a method for preparing the same, and an anti-cancer composition comprising the same derivative and, more specifically, to a pharmaceutical composition of dihydropyrazolecarbothioamide derivatives for anti-cancer which has an effect of inhibiting cell growth in human colon cancer cells.

Description

A dihydropyrazole carbothioamide derivative, a method for preparing the same, and an anticancer agent composition containing the derivative,

The present invention relates to an anticancer composition comprising a dihydropyrazole carbothioamide derivative, a process for producing the same, and derivatives thereof, and more particularly, to an anticancer composition comprising a dihydropyrazole carbothioamide derivative, To a pharmaceutical composition for an anticancer drug.

The simplest method for screening for substances showing anticancer effects is to measure whether or not cancer cells are killed by causing cytotoxicity against cancer cells. Since the anticancer effect is generally related to the structure of the substance, it is predicted that the difference in the anticancer effect will be large if the difference in structure is large. On the contrary, if the structural difference is similar, the difference in anticancer effect will be small. However, there is a colony forming assay (Clonogenic assay) method of cancer cells by screening method which clearly distinguishes the anticancer effect caused by a minute structural difference. This method has been widely used for screening substances with anticancer effects by analyzing the effect of specific substances on cell proliferation and survival [Franken NA, Rodermond HM, Stap J, Haveman J, van Bree C. Clonogenic assay cells in vitro. Nat Protoc . 2006; 1: 2315-9]. In addition, clonogenic survival assay method was used in the present invention in order to find a substance showing anticancer effect because it is widely used in recent years as a method of identifying differences in anticancer effects by microscopic structural differences.

To date, various anti-cancer drugs have been known and developed, but there remain problems to be solved. One of the ways to overcome this problem is the natural materials that have been used in civilian or traditional medicine for a long time. Plants produce secondary metabolites in small quantities for their defense, one of which is flavonoids. Since the flavonoid derivatives of mania species are already known, the design of flavonoid derivatives with new skeletons is absolutely necessary for the derivation of novel substances.

Korean Patent Laid-Open No. 10-2006-0033112 discloses that a composition comprising a flavonoid compound inhibits the activity of P-glycoprotein (P-gp), which is overexpressed in cancer cells, ), It can be used in medicine as an anticancer agent and anticancer agent because it increases cytotoxicity against cancer cells.

The present invention has been devised in view of the above needs, and an object of the present invention is to provide an effective anticancer agent.

Another object of the present invention is to provide an effective method for producing an anticancer agent.

In order to achieve the above object, the present invention provides a compound represented by the following general formula (1).

[Chemical Formula 1]

The present invention also provides a composition comprising, as an active ingredient, a compound of the following formula:

[Chemical Formula 1]

In Formula 1, R is selected from the group consisting of H, p-methoxy and o-methoxy.

The present invention also provides a pharmaceutical composition comprising, as an active ingredient, a compound represented by the following formula (1) and a pharmaceutically acceptable carrier:

[Chemical Formula 1]

In Formula 1, R is selected from the group consisting of H, p-methoxy and o-methoxy.

The present invention also provides an anticancer pharmaceutical composition comprising, as an active ingredient, a compound represented by the following formula (1) and a pharmaceutically acceptable carrier:

[Chemical Formula 1]

In Formula 1, R is selected from the group consisting of H, p-methoxy and o-methoxy.

In one embodiment of the present invention, the composition preferably has anticancer effect on colon cancer, but is not limited thereto.

The present invention also provides a process for preparing a benzochalcone compound, comprising the steps of: a) reacting 2'-hydroxy-1'-acetonaphthone with 2-methoxybenzaldehyde to obtain a benzocholacone compound;

b) reacting the benzocholactone compound with hydrazine to obtain 2-hydroxy-2-methoxy-pyrazoline, and c) reacting the 2-hydroxy-2-methoxy-pyrazoline and phenyl isothiocyanate phenyl isothiocyanate in the presence of a base.

[Chemical Formula 1]

In Formula 1, R is selected from the group consisting of H, p-methoxy and o-methoxy.

The present invention provides a pharmaceutical composition for preventing or treating a disease containing the compound of the formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient, wherein the disease is preferably cancer, and the composition comprising the compound of the present invention Can be preferably used for prevention or treatment of colorectal 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.

The compounds of formula (I) contained in the compositions of the present invention may also be used in the form of their salts, which salts include acid addition salts with various organic or inorganic acids which are pharmaceutically or physiologically acceptable. Suitable inorganic acids include, for example, hydrochloric acid, halogen acids such as sulfuric acid, and phosphoric acid. Suitable organic acids include, for example, carboxylic acid, phosphonic acid, sulfonic acid, acetic acid, propionic acid, octanoic acid, decanoic acid, glycolic acid, lactic acid, fumaric acid, succinic acid, adipic acid, malic acid, tartaric acid, citric acid, glutamic acid, , Maleic acid, benzoic acid, salicylic acid, phthalic acid, phenylacetic acid, benzenesulfonic acid, 2-naphthalenesulfonic acid, methylsulfuric acid, ethylsulfuric acid and dodecylsulfuric acid.

The composition of the present invention may further comprise a pharmacologically or physiologically acceptable carrier, excipient or diluent in addition to the compound of the formula (1) or a compound thereof.

Examples of suitable carriers, excipients and diluents that may be included in such a composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Methylcellulose, amorphous cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. The composition may further include conventional fillers, extenders, binders, disintegrants, surfactants, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers, preservatives and the like in the case of pharmaceutical formulation.

The composition of the present invention may be formulated into various forms such as powders, granules, tablets, capsules, oral formulations such as suspensions, emulsions, syrups and aerosols, and sterilized injection solutions according to conventional methods And can be administered via various routes including oral administration or intravenous, intraperitoneal, subcutaneous, rectal, topical administration, and the like.

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, lactose, gelatin, Mix and formulate. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used.

Examples of the oral liquid preparation include suspensions, solutions, emulsions, and syrups. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used.

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. Base materials for injections may include conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifying agents, stabilizers, and preservatives.

In the present invention, "administration" means providing a predetermined substance to a patient in any suitable manner, and the administration route of the composition of the present invention may be oral or parenteral ≪ / RTI > The composition may also be administered by any device capable of transferring the active agent to the target cell.

"Patient" in the present invention means an animal such as a human, a monkey, a dog, a goat, a pig, or a mouse having a disease in which symptoms can be improved by administering the composition of the present invention. The compositions according to the present invention are applicable not only to humans (treatment, inhibition or prevention) but also to other commercially useful animals.

In another aspect, the present invention provides a method for inhibiting, preventing and treating cancer and cancer metastasis by administering to a patient a composition comprising the compound of Formula 1 or a pharmaceutically acceptable salt thereof. The composition of the present invention can be administered in parallel with the above-mentioned conventional therapeutic agents for diseases.

The composition of the present invention is administered in a pharmaceutically effective amount.

In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level will depend on the type of disease, severity, , Sensitivity to the drug, time of administration, route of administration and rate of release, duration of treatment, factors including co-administered drugs, and other factors well known in the medical arts. The composition of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered singly or in multiple doses. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without side effects, which can be easily determined by those skilled in the art.

Further, the dosage of the compound according to the present invention may be varied depending on the degree of absorption, body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, severity of disease, However, for the desired effect, the compound of the present invention is preferably administered at 0.001 to 150 mg, preferably 0.01 to 100 mg per kg body weight per day. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.

The compound of formula (I) of the present invention can be synthesized by a method of synthesizing the reaction formula described in the present invention or various methods well known in the art.

Hereinafter, the present invention will be described.

The inventors of the present invention have devised a novel flavonoid derivative in which the C3 carbon skeleton is substituted with dihydropyrazole carbothioamide, and the anticancer effects of these compounds are measured using the clonogenic survival assay , And thus the present invention has been completed.

The present invention can be provided as a pharmaceutical composition for cancer prevention and anticancer drug by devising and synthesizing dihydropyrazole carbothioamide derivatives and observing the inhibitory effect on cell growth of colon cancer cells.

As can be seen from the present invention, the dihydropyrazole carbothioamide derivatives of the present invention are useful as pharmaceutical compositions for the prevention and treatment of cancer diseases by showing the cell growth inhibitory effect of colon cancer cells have.

1 is a diagrammatic representation of a dihydropyrazole carbothioamide derivative 3- (2-hydroxynaphthalen-1-yl) -5- (2-methoxyphenyl) -Carbothioamide ( DK111 ). ≪ / RTI > (Using a 400 MHz Bruker nuclear magnetic resonance spectrometer)
2 is a graph showing the results obtained when the dihydropyrazole carbothioamide derivative 3- (2-hydroxynaphthalen-1-yl) -5- (2-methoxyphenyl) Carbon < / RTI > amide ( DK111 ). (Using a 100 MHz Bruker nuclear magnetic resonance spectrometer)
FIG. 3 is a graph showing the results obtained when the dihydropyrazole carbothioamide derivative 3- (2-hydroxynaphthalen-1-yl) -5- (2-methoxyphenyl) -Carbothioamide ( DK111 ). ≪ / RTI > (Using a JMS700 HREIMS (high-resolution electron impact ionization mass spectrometer) instrument from Jeol Ltd., Tokyo, Japan)
FIG. 4 is a graph showing the effect of the dihydropyrazole carbothioamide derivative 3- (2-hydroxynaphthalene-1-yl) -5- (2-methoxyphenyl) -N- (4-methoxyphenyl) Hydrogen nuclear magnetic resonance spectroscopy of dihydropyrazole-1-carbothioamide ( DK112 ). (Using a 400 MHz Bruker nuclear magnetic resonance spectrometer)
FIG. 5 is a graph showing the effect of the dihydropyrazole carbothioamide derivative 3- (2-hydroxynaphthalene-1-yl) -5- (2-methoxyphenyl) -N- (4-methoxyphenyl) Carbon atom nuclear magnetic resonance spectroscopy of dihydropyrazole-1-carbothioamide ( DK112 ). (Using a 100 MHz Bruker nuclear magnetic resonance spectrometer)
FIG. 6 is a graph showing the activity of the dihydropyrazole carbothioamide derivative 3- (2-hydroxynaphthalen-1-yl) -5- (2-methoxyphenyl) -N- (4-methoxyphenyl) High-resolution mass spectrometry spectrum of dihydropyrazole-1-carbothioamide ( DK112 ). (Using a JMS700 HREIMS (high-resolution electron impact ionization mass spectrometer) instrument from Jeol Ltd., Tokyo, Japan)
7 is a graph showing the results of the synthesis of the dihydropyrazole carbothioamide derivative 3- (2-hydroxynaphthalen-1-yl) -N, 5-bis (2-methoxyphenyl) ≪ / RTI > is the hydrogen nuclear magnetic resonance spectroscopy spectrum of carbothioamide ( DK116 ). (Using a 400 MHz Bruker nuclear magnetic resonance spectrometer)
8 is a graph showing the results of the synthesis of the dihydropyrazole carbothioamide derivative 3- (2-hydroxynaphthalen-1-yl) -N, 5-bis (2-methoxyphenyl) -4,5-dihydropyrazol- Carbon < / RTI > nuclear magnetic resonance spectroscopy of carbothioamide ( DK116 ). (Using a 100 MHz Bruker nuclear magnetic resonance spectrometer)
FIG. 9 shows the effect of dihydropyrazole carbothioamide derivatives on cell growth in colon cancer cells.

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 examples are for illustrative purposes only and that the scope of the present invention is not construed as being limited by these examples.

Example  One. Dihydropyrazole carbothioamide  Synthesis of derivatives

In the present invention, the dihydropyrazole carbothioamide derivatives represented by formula (1) were synthesized as follows using the method shown in the following reaction formula.

(1, 864 mg, 10 mmol) and 2-methoxybenzaldehyde (1,360 mg, 10 mmol) were dissolved in 70 mL of ethanol and the temperature of the mixture was adjusted to 5 째 C Lower. Add 10 mL of 50% KOH solution slowly to the above cooling solution and stir at room temperature for about 30 hours. The reaction mixture is put into about 200 mL of ice water, acidified with 30 mL of 6N HCl, and the resultant solid is filtered. This solid was recrystallized from ethanol to obtain a pure benzocholacone compound ( I ). After dissolving the benzocholacone compound ( I , 1 mmol) and hydrazine (1.2 mmol) in ethanol (30 mL), the reaction mixture was refluxed for about 10 hours. The reaction mixture was reduced to 5 ° C. using an ice water bath. And the filtered solid was recrystallized in ethanol to synthesize a pure pyrazoline derivative II . Dissolved pyrazoline (II, 2-hydroxy4-methoxy pyrazoline, 950 mg, 3 mmol) and phenyl isothiocyanate (phenyl isothiocyanate 405 mg, 3 mmol ) in 10 ㎖ ethanol - 2-hydroxy-2-methoxy Thereafter, the mixture is stirred at reflux for 4 hours at 90 ° C. After cooling to room temperature, the resulting solid is washed with ethanol and filtered under reduced pressure. The solid thus formed was recrystallized in ethanol to obtain 1 g (78%) of the desired dihydropyrazole carbothioamide compound DK-111 .

Nuclear magnetic resonance spectroscopy experiments were performed to confirm the final product. The device used was a Bruscia 400MHz device. High resolution mass spectrometry was also used to confirm the structure of the derivatives identified by nuclear magnetic resonance spectroscopy. The instrument used was JMS700 HREIMS (high-resolution electron impact ionization mass spectrometer) from Jeol Ltd., Tokyo, Japan. Nuclear magnetic resonance spectroscopy confirmed the nomenclature according to the positions of hydrogen and carbon of the identifiers.

(2)

Dihydropyrazole carbothioamide derivative 3- (2-hydroxynaphthalen-1-yl) -5- (2-methoxyphenyl) -N-phenyl- Hydrogen Nuclear Magnetic Resonance Spectroscopy of the 3- (2-hydroxynaphthalen-1-yl) -5- (2-methoxyphenyl) -N-phenyl-4,5- dihydropyrazole- 1-carbothioamide DK111 ) Resonance spectra are as shown in FIGS. 1 and 2, respectively, and the chemical mobility is as follows.

^{1 H NMR (400MHz, DMSO-} d 6) δ 10.36 (bs, 1H, 2'-OH), 9.99 (s, 1H, 7-NH), 8.11 (d, 1H, H-8 ', J = 8.5 Hz , 7.87 (d, 1H, H-4 ', J = 8.9 Hz), 7.83 (d, 1H, H-5', J = 8.0 Hz) -6 ''', J = 7.5 Hz), 7.48 (ddd, 1H, H-7', J = 1.2, 6.9, 8.5 Hz), 7.33 (m, 1H, H-6 '), 7.31 (m, 2H , 7.26 (m, 1H, H-4 ''), 7.21 (d, 1H, H-3 ', J = 8.9 Hz), 7.16 (dd, 1H, H-6 '', J = 1.5, 7.6 Hz), 7.12 (t, 1H, H-4 ''', J = 7.5 Hz), 7.05 (d, 1H, H-3'', J = 8.1 Hz), 6.98 (t, 1H , H-5 '', J = 7.6 Hz), 6.25 (dd, 1H, H-5, J = 3.3, 11.3 Hz), 4.12 (dd, 1H, H-4, J = 11.3, 18.2 Hz), 3.85 (s, 3H, 2 '' - OCH 3), 2.98 (dd, 1H, H-4, J = 3.3, 18.2 Hz); ^{13 C NMR (100MHz, DMSO-} d 6) δ 173.8 (C-6), 156.7 (C-3), 155.7 (C-2 ''), 154.7 (C-2 '), 139.6 (C-1'''), 132.2 (C-9'), 131.4 (C-4 '), 129.6 (C-1''''/C-5'''), 127.7 (C-10 '), 127.3 (C-7'), 125.5 '), 124.6 (C-4'''), 123.9 (C-8 '), 123.1 (C-6'), 120.2 -3 ''), 110.8 (C -1 '), 58.9 (C-5), 55.5 (2''- OCH 3), 45.1 (C-4).

Dihydropyrazole carbothioamide derivative 3- (2-hydroxynaphthalen-1-yl) -5- (2-methoxyphenyl) -N-phenyl- The thioamide ( DK111 ) has a molecular formula of C ₂₇ H ₂₃ N ₃ O ₂ S as a new substance not reported to date. High-resolution mass spectrometry was used to confirm the structure of this compound as confirmed by nuclear magnetic resonance spectroscopy. Since the theoretical molecular weight was 453.1511 and the molecular weight obtained in the experiment was 453.1628, this compound was found to be 3- (2-hydroxynaphthalen- ) -5- (2-methoxyphenyl) -N-phenyl-4,5-dihydropyrazole-1-carbothioamide. The high-resolution mass spectrometry spectrum of this compound is shown in FIG.

Dihydropyrazole carbothioamide derivative 3- (2-hydroxynaphthalene-1-yl) -5- (2-methoxyphenyl) -N- (4-methoxyphenyl) (2-methoxyphenyl) -N- (4-methoxyphenyl) -4,5-dihydropyrazole-1-carbothioamide ( DK112 ) The hydrogen nuclear magnetic resonance spectroscopy and the carbon nuclear magnetic resonance spectrum are as shown in FIG. 4 and FIG. 5, respectively, and the chemical mobility is as follows.

^{1 H NMR (400MHz, DMSO-} d 6) δ 10.39 (bs, 1H, 2'-OH), 9.87 (s, 1H, 7-NH), 8.10 (d, 1H, H-8 ', J = 8.5 Hz ), 7.86 (d, 1H, H-4 ', J = 8.9 Hz), 7.83 (d, 1H, H-5', J = 8.0 Hz), 7.48 (ddd, 1H, H-7 ', J = 1.3 , 6.9, 8.5 Hz), 7.42 (d, 2H, H-2 '''/H-6''', J = 9.0 Hz), 7.32 (ddd, 1H, H-6 ', J = 1.0, 6.9, 8.0 Hz), 7.27 (m, 1H, H-4 ''), 7.20 (d, 1H, H-3 ', J = 8.9 Hz), 7.15 (dd, 1H, H-6'', J = 1.5, 7.6 Hz), 7.05 (d, 1H, H-3 '', J = 8.1 Hz), 6.98 (t, 1H, H-5 '', J = 7.6 Hz), 6.87 (d, 2H, H-3 ''' / H-5 ''', J = 9.0 Hz), 6.23 (dd, 1H, H-5, J = 3.4, 11.3 Hz), 4.11 (dd, 1H, H-4, J = 11.3, 18.1 Hz ), 3.84 (s, 3H, 2 '' - OCH 3), 3.74 (s, 3H, 4 '''- OCH 3), 2.95 (dd, 1H, H-4, J = 3.4, 18.1 Hz); ^{13 C NMR (100MHz, DMSO-} d 6) δ 173.3 (C-6), 156.5 (C-4 '''), 156.4 (C-3), 155.7 (C-2''), 154.7 (C-2 '), 132.6 (C-1'''), 132.1 (C-9 '), 131.3 (C-10 '), 127.2 (C-7'), 126.9 (C-2 '' / C-6 ''), 125.5 (C-6 '), 120.2 (C-5''), 118.1 (C-3'), 113.1 (C-3 ' -3 ''), 110.8 (C -1 '), 58.9 (C-5), 55.5 (2''- OCH 3), 55.1 (4''' - OCH 3), 45.1 (C-4).

Dihydropyrazole carbothioamide derivative 3- (2-hydroxynaphthalene-1-yl) -5- (2-methoxyphenyl) -N- (4-methoxyphenyl) -l- carbonyl thioamide (DK112) has a molecular formula of _{C 28 H 25 N 3 O 3} S as a new material that is not reported to date. High-resolution mass spectrometry was used to confirm the structure of this compound as confirmed by nuclear magnetic resonance spectroscopy. Since the theoretical molecular weight was 483.1617 and the molecular weight obtained in the experiment was 483.1717, the compound was found to be 3- (2-hydroxynaphthalen-1-yl ) -5- (2-methoxyphenyl) -N- (4-methoxyphenyl) -4,5-dihydropyrazole-1-carbothioamide. The high-resolution mass spectrometry spectrum of this compound is shown in Fig.

Dihydropyrazolecarbothioamide derivative 3- (2-hydroxynaphthalen-1-yl) -N, 5-bis (2-methoxyphenyl) -4,5-dihydropyrazole- Amide (3- (2-hydroxynaphthalen-1-yl) -N, 5-bis (2-methoxyphenyl) -4,5- dihydropyrazole-1-carbothioamide DK116 ) and carbon nuclear magnetic resonance spectroscopy Are shown in FIGS. 7 and 8, respectively, and the chemical mobility thereof is as follows.

¹ H NMR (400 MHz, DMSO-d ₆ )? 10.48 (bs, 1H, 2'-OH), 9.87 (d, 1H, H-8 ', J = 8.6 Hz), 7.89 (d, 1H, H-4', J = 8.9 Hz), 7.86 (d, 1H, H-5 ', J = 8.0 Hz), 7.53 (ddd, 1H, H- 7 ', J = 0.9, 6.9, 8.6 Hz), 7.37 (ddd, 1H, H-6', J = 0.9, 6.9, 8.0 Hz), 7.25 (m, 1H, H- 1H), 7.01 (m, 1H, H-6 ''), 7.21 (d, 1H, H-3 ', J = 8.9 Hz), 7.10 (m, 1H, H-3 ''), 7.06 (m, 1H, H-3 '''), 6.94 '), 6.20 (dd, 1H , H-5, J = 3.3, 11.3 Hz), 4.14 (dd, 1H, H-4, J = 11.3, 18.2 Hz), 3.85 (s, 3H, 2''- OCH _{3), 3.80 (s, 3H} , 2 '''- OCH 3), 3.15 (dd, 1H, H-4, J = 3.3, 18.2 Hz); ^{13 C NMR (100MHz, DMSO-} d 6) δ 172.1 (C-6), 156.8 (C-3), 155.7 (C-2 ''), 155.5 (C-2 '), 150.0 (C-2'''), 132.0 (C-9'), 131.9 (C-4 '), 129.3 (C-1''), 128.4 '''), 127.9 (C-10'), 127.4 (C-7 '), 125.6 C-6 '), 122.0 (C-6'''), 120.2 (C-5 ''), 119.7 ), 110.9 (C-3 '''), 110.2 (C-1'), 58.3 (C-5), 56.0 (2 '''- OCH 3), 55.5 (2''- OCH 3), 45.4 ( C-4).

Dihydropyrazolecarbothioamide derivative 3- (2-hydroxynaphthalen-1-yl) -N, 5-bis (2-methoxyphenyl) -4,5-dihydropyrazole- amide (DK116) has a molecular formula of _{C 28 H 25 N 3 O 3} S as a new material that is not reported to date. The structure of this compound confirmed by nuclear magnetic resonance spectroscopy was unnecessary for further high resolution mass spectrometry experiments as compared to the structures of DK111 and DK112 mentioned above.

Example  2. Dihydropyrazole carbothioamide  Inhibitory effect of derivatives on cell growth of colon cancer cells

HCT116 colon cancer cells were purchased from the American Type Culture Collection (ATTC) and DMEM (Invitrogen Life Technologies) culture medium containing 10% FBS (Fetal Bovine Serum, Invitrogen Life Technologies) and Antibiotic-Antimycotic solution The cells were cultured in a 5% CO ₂ incubator at 37 ° C in a 100-mm cell culture dish at a seeding density of 1 × 10 ⁶ . The inhibitory effect of the benzohydroxymethoxycalcone compound (DK49) on cell proliferation was determined by whether the cell growth was inhibited by a colony forming assay of cancer cells. HCT116 colon cancer cells were divided into 6000 cells per well in a 24-well culture dish, treated with dihydropyrazole carbothioamide derivatives at concentrations of 0, 5, 10 and 20 μM, and after 7 days, 6% glutar A mixture of aldehyde (0.5%) and crystal violet (1: 1) was added to the cells, followed by reaction for 15 minutes to stain the remaining cells. As a result, it was observed that when the dihydropyrazole carbothioamide derivative was treated at a concentration of 5 μM or more as shown in FIG. 9, the colony forming ability of the cancer cells was drastically reduced. From these facts, it has been confirmed that the dihydropyrazole carbothioamide derivative compound of the present invention has an effect of inhibiting the growth of 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 (6)

A compound of formula

[Chemical Formula 1]
In Formula 1, R is selected from the group consisting of H, p-methoxy and o-methoxy. A composition comprising a compound of the formula (1) as an active ingredient:

[Chemical Formula 1]
In Formula 1, R is selected from the group consisting of H, p-methoxy and o-methoxy. A pharmaceutical composition comprising, as an active ingredient, a compound represented by the following formula (1) and a pharmaceutically acceptable carrier:

[Chemical Formula 1]
In Formula 1, R is selected from the group consisting of H, p-methoxy and o-methoxy. An anticancer pharmaceutical composition comprising a compound represented by the following formula (1) and a pharmaceutically acceptable carrier as an active ingredient:

[Chemical Formula 1]
In Formula 1, R is selected from the group consisting of H, p-methoxy and o-methoxy. 5. The anticancer pharmaceutical composition according to claim 4, wherein the composition has anticancer effect on colon cancer. a) reacting 2'-hydroxy-1'-acetonaphthone with 2-methoxybenzaldehyde to obtain a benzalkonium compound;
b) reacting the benzocholactone compound with hydrazine to obtain 2-hydroxy-2-methoxy-pyrazoline; and
c) reacting the 2-hydroxy-2-methoxy-pyrazoline with phenyl isothiocyanate.

[Chemical Formula 1]
In Formula 1, R is selected from the group consisting of H, p-methoxy and o-methoxy.

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