KR101540033B1 - A novel benzochalcones and composition for anticancer comprising the same - Google Patents

Abstract

The present invention relates to a novel benzochalcone derivative and an anticancer composition comprising the compound.

Description

TECHNICAL FIELD The present invention relates to novel benzochalcone derivatives and their use as an anticancer composition,

The present invention relates to a novel benzochalcone derivative and an anticancer composition comprising the compound.

Colon cancer is a malignant tumor consisting of cancer cells that develop in the large intestine.

When we eat food, the ingested food is excreted in the feces through the digestive tract. The digestive organs of our body are divided into the esophagus, stomach, small intestine, and large intestine. The large intestine is the last part of the digestive tract and mainly absorbs water and electrolytes. The large intestine is divided into the colon and the rectum. The colon is divided into the cecum, ascending colon, transverse colon, descending colon and S (S) colon. The cancer that develops in the colon depends on colon cancer, rectum Is referred to as rectal cancer, collectively referred to as colon cancer or colorectal cancer. The incidence of cancer in each part of the large intestine is known to be about 25% in the cecum and ascending colon, 15% in the transverse colon, 5% in the descending colon, 25% in the S colon, 10% in the rectum-S colon interface and 20% in the rectum.

The large intestine is a pipe-shaped tube divided into four layers from the inside, including mucosal layer, submucosal layer, muscle layer, and serous layer. Most colorectal cancers are adenocarcinomas arising in the mucosa of the large intestine. In addition, there are lymphoma, sarcoma, squamous cell carcinoma, metastatic lesions of other cancers.

5-Fluorouracil (5-FU) has been used for many years in the treatment of colorectal cancer. And is typically used in combination with leucovorin to increase its effect [C. Emiliano, Clin. Colorectal Canc. 2 (2002) 170-172). Chemotherapeutic agents such as Camptosar, Eloxatin, Avastin, Erbitux, and Vectibix have been developed and used in the treatment of colorectal cancer, but they also affect healthy cells as well as target cancer cells [T. Cartwright, Clin. Colorectal Canc. (2011) doi 10.1016 / j.clc.2011.11.001]. Therefore, chemotherapy drugs for colon cancer are still being developed.

Chalcone are plant secondary metabolites and belong to the flavoid class with the C6-C3-C6 backbone (Figure 1A). Unlike most flavonoids, however, chalcone does not have a closed ring in their middle structure (C-ring), as shown in Figure 1B. They are made by chalcone synthase in plant biosynthetic pathways or synthesized from aldol condensation of acetophenone and benzaldehyde.

[Prior Patent Literature]

Korean Patent Publication No. 10-2013-0044837

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

In order to accomplish the above object, the present invention provides a process for the preparation of 1- (5- (2,4-dimethoxyphenyl) -4,5-dihydro-1H-pyrazol-3-yl) naphthalen- ).

The present invention also provides a pharmaceutical composition for anticancer therapy comprising the compound of the present invention as an active ingredient.

The present invention also provides a composition for stopping the cell cycle comprising the compound of the present invention as an active ingredient.

The present invention also provides a composition for inducing epitope in which the compound of the present invention is contained as an active ingredient.

The present invention also relates to a process for the production of benzoquinone derivatives by dissolving 2'-hydroxy-1'-acetonaphthone and 2,4-dimethoxy substituted benzaldehyde in ethanol and reacting them, And treating the benzochalcone derivative with hydrazine. The present invention also provides a method for producing the compound of the present invention.

The composition of the present invention has an effect on 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.

The pharmaceutical compositions containing the compounds of the present invention may be formulated in the form of powders, granules, tablets, capsules, oral preparations such as suspensions, emulsions, syrups and aerosols, external preparations, suppositories or sterile injection solutions, Can be used. In detail, when formulating the composition, it can be prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, etc. which are usually used.

The solid preparation for oral administration can be prepared by mixing at least one excipient such as starch, calcium carbonate, sucrose, lactose, gelatin and the like in the compound of the present invention. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterile 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 withexol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The preferred dosage of the compound of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. However, for a desired effect, the compound of the present invention or a pharmaceutically acceptable salt thereof may be administered in an amount of 0.0001 to 100 /, preferably 0.001 to 100 /, divided once to several times per day. In a total pharmaceutical composition, the compound of the present invention or a pharmaceutically acceptable salt thereof should be contained in an amount of 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, and the like in various routes. The administration route can be administered by various methods such as oral, rectal, intravenous, .

Hereinafter, the present invention will be described.

The present inventors report on the synthesis of benzochalcone having 17 novel pyrazoles and their anticancer activities.

In order to find a powerful and novel chemotherapeutic agent for colorectal cancer, a series of benzochalcone derivatives (the alpha, beta unsaturated carbonyl group substituted with pyrazoline) were devised and synthesized. A colony forming survival assay was performed with each derivative to assess anticancer activity.

Naphthalen-2-ol (derivative 7) is a potent inhibitor of long-term colony formation of HCT116 human colon cancer cells. Effect. The results of western blot and flow cytometry suggest that derivative 7 may inhibit the proliferation of colon cancer cells through inhibition of cell cycle progression and induction of epopotisis.

A general route to the synthesis of benzochalcones 1-17 is shown in Fig. The Claisen-Schmidt condensation of 2'-hydroxy-1'-acetonaphthone (I) with substituted methoxy benzaldehydes (II) in base conditions provides benzochalcones (III). The benzochalcones (III) were treated with p-chlorophenyl hydrazine (IV) under ethanol reflux to yield novel 1,3,5-trisubstituted benzochalcones 1-4 in good yield. The benzochalcones (III) were reacted with hydrazine (V) to give novel 3,5-disubstituted benzochalcones 5-8. Benzochalcones (VI) were obtained by Claisen-Schmidt condensation of 1'-hydroxy-2'-acetonaphthone with substituted methoxy benzaldehydes (II). The benzochalcones (VI) were then treated with hydrazine to produce the novel 3,5-disubstituted benzochalcones 9-17. The chemical identification of benzochalcone derivatives 1-17 was performed based on the analysis of basic one-dimensional and two-dimensional NMR spectroscopy and high resolution electron impact ionization mass spectrometry (HREIMS).

Derivative 1 The procedure used to assign the NMR data of 1- (1- (4-chlorophenyl) -5- (4-methoxyphenyl) -4,5-dihydro-1H- pyrazol- Will be described in detail. Of the 26 carbon atoms of derivative 1, the most shielded carbon atom is C-4. Thus, at 48.8 ppm, the ¹³ C peak is assigned to C-4, which is directly attached to the two protons observed at 3.68 and 4.31 ppm in the heteronuclear multiple quantum coherence (HMQC) spectrum 8). Based on the COZY (correlated spectroscopy) spectrum, H-5 can be assigned to the peak at 5.21 ppm (FIG. 9). H-4 represents two long-range couplings with two singlet carbon atoms at 150.5 and 132.7 ppm in the heteronuclear multiple quantum coherence (HMQC) spectrum (Fig. 10). The front peak did not represent another long-range coupling with protons, so we assigned it to C-3. In conclusion, the latter peak is assigned to C-1 '', which represents a long-range coupling with a ¹ H peak at 6.88 ppm. At 114.7 ppm, the ¹³ C peak suggests a direct attachment to the 6.88 ppm protons, which correlates with the ¹ H peak at 7.24 ppm in the COZY spectrum. This proton is attached to the carbon atom that participates in the peak at 127.2 ppm. At 114.7 and 127.2 ppm, since the ¹³ C peak is twice the intensity of the adjacent carbon signal (Fig. 11), they are respectively at atoms C-3 '' / C-5 '' and C-2 '' / C-6 '' Respectively. At 6.88 ppm, the ¹ H peak exhibits a nuclear overhauser effect (nOe) cross peak with a ¹ H peak at 3.78 ppm attached to the ¹³ C peak at 55.3 ppm in the nuclear Overhauser Effect Spectroscopy (NOESY) spectrum 12). The ¹³ C peak at 55.3 ppm is a signal caused by 4 "-OCH ₃ , since it has only one methoxy carbon in this compound. The methoxy proton is coupled to the ¹³ C peak and the long-range at 159.4 ppm, -4 ". At 114.8 and 129.1 ppm, the signals are assigned to C-2 '' / C-6 '' and C-3 '' / C-5 '', respectively, twice the adjacent signals in the ¹³ C NMR spectrum. The yielding peaks at 6.93 and 7.16 ppm are attached to C-2 '' / C-6 '' and C-3 '' / C-5 ''' And H-3 '''/H-5''. Long-range coupling is observed at two additional carbon peaks at 125.0 and 142.6 ppm, which are assigned to C-4 '''andC-1''', respectively. ¹³ C peaks at 109.1 ppm represent H-4 and long-range coupling and are assigned to C-1 ', which is coupled to two proton peaks at 7.27 and 7.92 ppm. These peaks are assigned to H-3 'or H-8'. The nOe cross peak was observed between the two protons at 12.11 and 7.27 ppm. At 12.11 ppm, the ¹ H peak is attributed to 2'-OH, so at 7.27 ppm, the ¹ H peak is assigned to H-3 '(Figure 13). Thus, at 7.92 ppm, a ¹ H peak is assigned to H-8 '. The COZY spectrum also allows assignment of H-5 ', H-6', and H-7 '. At 131.9 ppm, the undetermined doublet carbon is assigned to C-4 '. At 157.4 ppm, the ¹³ C peak is assigned to C-2 'since it couples to H-4' with a long-range. At 131.9 ppm, the singlet anti-carbon peaks represent three long-range couplings with H-4 ', H-5', and H-7 'and are therefore due to C-9'. H-5 represents two ³ J values, 7.6 Hz and 11.3 Hz. cyclopentene ³ J _trans is smaller than ³ J _cis , so H-4 with H-4 with δ = 3.68 ppm and ³ J = 7.6 Hz is assigned to H-4 _trans relative to H-5.

Hereinafter, the present invention will be described in detail.

17-benzochalcone having a pyrazoline moiety at the position of an α, β-unsaturated carbonyl group was synthesized and 1- (4-chlorophenyl) -5-methoxyphenyl-4,5-dihydro-1H- pyrazol- 2-ols (Fig. 2A), 1- (5-methoxyphenyl-4,5-dihydro-1H- pyrazol-3-yl) naphthalen- -4,5-dihydro-1H-pyrazol-3-yl) naphthalen-1-ols (FIG. 2C). The first group consists of derivatives 1-4, the second group contains 5-8, and the last group contains derivatives 9-17. These derivatives differ only in the position and number of methoxy groups, and their biological properties can not be distinguished by structural analysis alone. The measurement of cell viability for anticancer agents is widely used for screening biological activity. The most common method of measuring short-term loss of cell viability such as MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) . However, clonogenic cell survival assays (CSAs) detect long-term cell proliferation inhibitory effects caused by anticancer drugs. CSAs can be used as a dose-dependent index of cell viability [P.R. Roper, B. Drewinko, Cancer Res. 36 (1976) 2182-2188]. Thus, in order to assess the biological activity of 17 derivatives, a colony forming survival assay was applied using HCT116 human colon cancer cells. Compounds that exhibit complete inhibition in a colony forming assay give rise to zero cell viability. Therefore, the presence of chlorophenyl groups reduces the inhibitory effect.

The only structural difference between the first and second groups is the presence and absence of chlorophenyl groups, respectively. Derivatives 1 and 5 contain the same substituent (4 " -methoxy group), and derivatives 2 and 7 both contain 2 ", 4 " -dimethoxy groups. The cell survival rates as measured by the colony forming survival assay for derivatives 1 and 5 were 83.7 and 72.3, respectively. Similarly, cell viability for derivatives 2 and 7 were 52.2 and 42.4, respectively.

The only structural difference between the second and third groups is the relative position of the hydroxy and pyrazoline groups. Derivatives 6 and 9 contain the same 2 ", 3 " -dimethoxy substituent and derivatives 8 and 14 have the 3 ", 5 " -dimethoxy group. The cell survival rates of derivatives 6 and 9 were 65.2 and 108.8, respectively. Similarly, the cell viability of derivatives 8 and 14 were 58.3 and 58.4, respectively. Thus, the 1-pyrazolylnaphthalen-2-ol moiety showed greater inhibitory effect than the 2-pyrazolylnaphthalen-1-ol moiety.

The cell viability of derivatives 3 (57.0) and 4 (70.2) was lower than that of 15 (82.5) and 16 (73.3), respectively. Thus, with respect to colony formation, the relative positions of the hydroxy and pyrazoline groups are more important than the presence of chlorophenyl groups. Among 17 kinds of benzochalcone tested in the present invention, derivatives 7 and 17 showed the highest inhibitory effect and the lowest inhibitory effect of 42.4 and 192.6, respectively (Fig. 3A). The inhibition values of 17 derivatives obtained from the densitometric analysis are shown in Fig. 3B. The strongest and weakest inhibitory effects were 85.9% and 35.8% inhibition, respectively, and were 1- (5- (2,4-dimethoxyphenyl) -4,5-dihydro-1H- pyrazol-3-yl) naphthalen- Was induced by 2- (5- (2,4,6-trimethoxyphenyl) -4,5-dihydro-1H-pyrazol-3-yl) naphthalen-1-ol.

To further investigate the antitumor activity of derivative 7 which exhibited the greatest inhibition, exponentially growing HCT116 cells were exposed to various concentrations of derivative 7 at concentrations of 0 to 40 [mu] M for 24 to 48 hours. Treatment of derivative 7 resulted in inhibition of the growth rate in a dose- and time-dependent manner (Figure 4). Many anticancer drugs inhibit proliferation of tumor cells through inhibition of cell cycle progression. In order to determine the molecular mechanisms by which derivative 7 inhibits cell proliferation, we investigated the effect of derivative 7 on the expression of cell cycle regulating proteins in HCT116 cells. Western blot analysis indicated that the level of p21, an endogenous inhibitor of cyclin-dependent protein kinase (CDK), gradually increased in a time-dependent manner, while cyclin B1 levels decreased rapidly after 24 hours of treatment (Fig. 5). The level of cyclin D1 slightly increased within 12 hours, but then gradually returned to the control level. The level of cyclin A1 slightly changed. These data suggest that derivative 7 has an inhibitory effect on cell cycle progression, at least in part, through alteration of p21 and cyclin B1 expression. To confirm this effect, the HCT116 cell cycle profile was measured with the treatment of 40 [mu] M derivative 7. Flow cytometry showed that the group of G1 cells increased slightly at the same time as the decrease in the percentage of S and G2 / M stage cells (Fig. 6). After 48 hours, the epoptoctic sub-G1 cells showed 21.3% of the total cells. Taken together, these data suggest that inhibition of colony formation by derivative 7 is due to the regulation of cell cycle progression and induction of epopotisis.

As can be seen from the present invention, the results of the HCT116 colon cancer cell lines of each derivative showed that the 1-pyrazolylnaphthalen-2-ol substituent showed greater inhibition than the 2-pyrazolylnaphthalen-1-ol moiety and the hydroxy and pyrazoline group Relative location indicated that the presence of chlorophenyl group is more important. The HCT116 colon cancer cell proliferation, as determined by this colony forming assay, was measured as described in Example 1, It was the strongest inhibitor against. Additional biological results indicated that derivative 7 inhibited cell cycle progression and induced apoptosis through negative regulation of the cell cycle control proteins p21 and cyclin B1. These results indicate the possibility of developing a novel benzoquercone having a pyrazoline moiety for use as an anticancer drug.

1 shows the structure of (A) flavone, (B) chalcone, (C) 3,5-diphenyl-4,5-dihydro-1H-pyrazole and (D) 7,8-benzoflavone.
2-ol, (B) 1- (5-methoxyphenyl-4,5-dihydro-1H-pyrazol-3-yl) naphthalen- 4,5-dihydro-1H-pyrazol-3-yl) naphthalen-2-ol and (C) 2- (5-methoxyphenyl- Structure and Names of.
Figure 3 shows the effect of chalcone derivatives on the colony formation of HCT116 cells. HCT116 cells (5 × 10 ³ cells / well) were plated and cultured for 7 days in the presence or absence of each derivative at different concentrations. Similar results were obtained from two independent experiments. (A) the largest inhibitory effect was observed in derivative 7 and the lowest inhibitory effect was observed in derivative 17. The values obtained by densitometric analysis are shown in (B).
Figure 4 shows the effect of derivative 7 on the proliferation of HCT116 cells. HCT116 cells were treated with another dose (0, 5, 10, 20, or 40 [mu] M) of derivative 7 for 24 or 48 hours.
Figure 5 shows the effect of derivative 7 on the expression of cell cycle regulatory proteins. HCT116 cells were treated with 40 [mu] M derivative 7 for the time indicated. Whole cell lysate was prepared and loaded on 10% SDS-PAGE. After electrophoresis, the proteins were transferred to a blot and immunoreacted with the described antibody. GAPDH was used as an internal loading control. Relative band intensities were measured with a quantitative scanning densitometer and the values were averaged ± standard standard deviation.
6 shows the effect of the cell cycle progression of the derivative 7. Fig. HCT116 cells were treated for 40 min at a time indicated by derivative 7 and analyzed for the amount of propidium iodide-stained DNA by flow cytometry. The x-axis represents the percentage of cells in the cell cycle. The quantities of 2N (diploid) and 4N (diploid) DNA represent the G1 and G2 / M stages of the cell cycle, respectively. At the sub-G1 stage, the cell population represents cell division due to epopotis. M1, sub-G1 step; M2, G1 step; M3, step S; M4, G2 / M steps. Values are expressed as mean ± one standard deviation of three independent experiments.
Figure 7 is a synthetic method used in the preparation of benzochalcone derivatives 1-17.
8 shows the HMQC spectrum of the derivative 1, 1- (1- (4-chlorophenyl) -5- (4-methoxyphenyl) -4,5-dihydro-1H- pyrazol-3-yl) naphthalene-2-ol.
9 is a COZY spectrum of the derivative 1, 1- (1- (4-chlorophenyl) -5- (4-methoxyphenyl) -4,5-dihydro-1H- pyrazol-3-yl) naphthalene-2-ol.
10 shows the HMBC spectrum of the derivative 1, 1- (1- (4-chlorophenyl) -5- (4-methoxyphenyl) -4,5- dihydro-1H- pyrazol-3- yl) naphthalene-2-ol.
11 is a ¹³ C NMR spectrum of the derivative 1, 1- (1- (4-chlorophenyl) -5- (4-methoxyphenyl) -4,5-dihydro-1H- pyrazol-3-yl) naphthalene-2-ol.
12 is a NOESY spectrum of the derivative 1, 1- (1- (4-chlorophenyl) -5- (4-methoxyphenyl) -4,5-dihydro-1H- pyrazol-3-yl) naphthalene-2-ol.
13 shows the ¹ H NMR spectra of the derivative 1, 1- (1- (4-chlorophenyl) -5- (4-methoxyphenyl) -4,5- dihydro-1H- pyrazol-3-yl) naphthalene-
Figure 14 shows the results of a colony formation survival assay of 17 different derivatives of three different concentrations (10, 20, and 40 [mu] M).

The present invention will now be described in more detail by way of non-limiting examples. The following examples are intended to illustrate the invention and the scope of the invention is not to be construed as being limited by the following examples.

All NMR experiments of the present invention were performed at 298 K with a Bruker Avance 400 spectrometer system (9.4 T; Bruker, Karlsruhe, Germany). Synthetic benzochalcon derivatives except derivatives 1, 3, and 4 were dissolved in DMSO-d ₆ . 1, 3, and 4 were dissolved in CHCl ₃ -d. The ¹ H and ¹³ C chemical shifts of deuterated substituted solvents were referenced to TMS (tetramethylsilane). NMR samples were prepared to about 50 mM and transferred to a 2.5-mm NMR tube. For one-dimensional ¹ H and ¹³ C NMR experiments, the relaxation delays are 1 s and 3 s with 90 ° pulses of 11.8 μs and 15.0 μs, respectively. The ¹ H and ¹³ C spectral widths were 4,800 and 21,000 Hz, respectively. For a two-dimensional experiment involving correlated spectroscopy (COZY), total correlation spectroscopy (TOCSY), nuclear overhauser effect spectroscopy (NOESY), HMQC and HMBC, all data are stored in 2K x 256 data points (t ₂ xt ₁ ). The sewing time for the NOESY experiment was 1 s and the long-range coupling time for the HMBC experiment was 70 ms. Zero-filling and sine-squared bell window functions of Zero-filling of 2 K were applied before Fourier transformation using XWin-NMR (Bruker) [G. Jo, J. Hyun, D. Hwang, YH Lee, D. Koh, Y. Lim, Magn. Reson. Chem. 49 (2011) 374-377). All NMR data were analyzed using Sparky [TD Goddard, DG Kneller, SPARKY 3, University of California, San Francisco (USA) (source: http://www.cgl.ucsf.edu/home/sparky/)] . All mass spectra were collected on a high resolution electron impact ionization mass spectrometer (HREIMS, JMS700, Jeol Ltd., Tokyo, Japan).

Example  1: General procedure for the synthesis of derivatives 1-8

2'-Hydroxy-1'-acetonaphthone (1,864 mg, 10 mmol) and methoxy-substituted benzaldehyde (10 mmol) were dissolved in 85 mL of ethanol and the temperature of the reaction mixture was adjusted to 5 DEG C in a cooling bath. To the cooled reaction mixture was added a solution of 50% KOH (10 mL) and the mixture was stirred at ambient temperature for 40 h. The reaction mixture was acidified by the addition of 6 N HCl (30 mL) to form a precipitate and re-crystallized with ethanol. When the acidification did not produce a precipitate, the aqueous layer was extracted twice with dichloromethane. The combined aqueous layer was dried them with MgSO _4. Filtration and distillation of the solvent gave a residue which was purified by flash chromatography (ethyl acetate: hexane = 1: 4) to give the corresponding benzochalcones (III). Benzochalcones (III, 1 mmol) and 4-chlorophenylhydrazine hydrochloride (1 mmol) were dissolved in ethanol (30 mL) and the reaction mixture was refluxed for 10 h. The temperature of the reaction mixture was adjusted to 5 캜 in a cooling bath to form a precipitate. Filtration and recrystallization gave pure derivatives 1-4. Benzochalcones (III, 1 mmol) was treated with hydrazine (1.2 mmol) to give derivatives 5-8.

Synthesis of 1- (1- (4-chlorophenyl) -5- (4-methoxyphenyl) -4,5-dihydro-1H- pyrazol-3-yl) naphthalen-

Color: Pale yellow; mp 161-163 [deg.] C; Yield 76%; ^{1 H NMR (400 MHz, CHCl} 3 -d) δ12.11 (s, 1H, 2'-OH), 7.92 (d, 1H, H-8 'J = 8.5 Hz), 7.77 (d, 1H, H- 1H, J = 8.0 Hz), 7.75 (d, 1H, H-4 'J = 9.0 Hz), 7.40 (ddd, H-6 'J = 1.2, 6.9, 8.0 Hz), 7.27 (d, 1H, H-3' 8.8 Hz), 7.16 (d, 2H, H-3 '''' H-5 '' J = 8.9 Hz), 6.93 (d, 2H, H- 8.9 Hz), 6.88 (d, 2H, H-3 '' H-5 '', J = 8.8 Hz), 5.21 (dd, 1H, H-4, J = 11.3, 16.7 Hz), 3.78 (s, 3H, 4 '' - OCH 3), 3.68 (dd, 1H, H-4, J = 7.6, 16.7 Hz); ^{13 C NMR (100 MHz, CHCl} 3 -d) δ159.4 (C-4 ''), 157.4 (C-2 ', 150.5 (C-3), 142.6 (C-1''', 132.7 (C- 1 ''), 131.9 (C-4 ', 131.9 (C-9', 129.4 (C-5 ', 129.1 (C-2 '' / C-6 ''), 126.8 (C-7 ', 125.0 , 114.8 (C-2 '''/C-6''', 114.7 (C-3 '' / C-5 ''), 109.1 '-OCH ₃ ), 48.8 (C-4); HREIMS (m / z): Calcd. For C ₂₆ H ₂₁ ClN ₂ O ₂ (M) ⁺ : 428.9101; Found 428.1295.

Synthesis of 1- (1- (4-chlorophenyl) -5- (2,4-dimethoxyphenyl) -4,5-dihydro-1H- pyrazol-3- yl) naphthalene-2-ol

Color: Pale yellow; mp 164 [deg.] C; Yield 69%; ^{1 H NMR (400 MHz, DMSO} -d 6) δ10.23 (bs, 1H, 2'-OH), 8.26 (d, 1H, H-8 'J = 8.6 Hz), 7.82 (d, 1H, H- 1H, J = 8.0 Hz), 7.81 (d, 1H, H-4 'J = 9.0 Hz), 7.47 (ddd, H-6 'J = 1.0, 6.9, 8.0 Hz), 7.23 (d, 1H, H-3', J = 9.0 Hz), 7.19 (d, 2H, H-3 ' = 9.0 Hz), 7.05 (d, 1H, H-6 '', J = 8.5 Hz), 6.89 (d, 2H, H-2 ''' 1H, H-3 '', J = 2.3 Hz), 6.48 (dd, 1H, H-5 '', J = 2.3, 8.5 Hz), 5.51 12.0 Hz), 4.02 (dd, 1H, H-4, J = 12.0, 17.6 Hz), 3.87 (s, 3H, 2 '' - OCH 3), 3.73 (s, 3H, 4 '' - OCH 3), 3.08 (dd, 1H, H-4, J = 12.0, 17.6 Hz); ^{13 C NMR (100 MHz, DMSO} -d 6) δ 159.9 (C-4 ''), 157.2 (C-2 ''), 154.3 (C-2 '), 148.2 (C-3), 143.6 (C- 1 ''', 132.6 (C-9'), 130.4 (C-4 '), 128.7 (C-3''), 127.1 (C-6''), 126.9 (C-7'), 124.1 (C-8 '), 122.9 ''), 118.2 (C-3 '), 113.9 (C-2''' / C-6 ''', 112.0 ''), 57.1 (C- 5), 55.7 (2 '' - OCH 3), 55.2 (4 '' - OCH 3), 46.0 (C-4); HREIMS (m / z):. Calcd for C 27 H ₂₃ ClN ₂ O ₃ (M) ⁺ : 458.9361; Found 458.1395.

Synthesis of 1- (1- (4-chlorophenyl) -5- (2,3,4-trimethoxyphenyl) -4,5-dihydro-1H- pyrazol-3-yl) naphthalen-

Color: Yellow; mp 166-168 [deg.] C; Yield 75%; ¹ H NMR (400 MHz, CHCl ₃ -d)? 12.07 (s, IH, 2'-OH), 7.96 J = 8.0 Hz), 7.75 (d, 1H, H-4 'J = 8.9 Hz), 7.42 (ddd, 1H, J = 1.0, 6.9, 8.0 Hz), 7.27 (d, 1H, H-3 'J = 8.9 Hz), 7.18 (d, 2H, H- J = 8.8 Hz), 6.84 (d, 1H, H-6 ', J = 8.6 Hz), 6.56 (d, 2H, (Dd, 1H, H-5 '', J = 8.6 Hz), 5.49 (dd, 1H, H-5, J = 6.7, , 3.99 (s, 3H, 3 '' - OCH 3), 3.88 (s, 3H, 2 '' - OCH 3), 3.81 (s, 3H, 4 '' - OCH 3), 3.61 (dd, 1H, H -4, J = 6.7, 16.7 Hz); ^{13 C NMR (100 MHz, CHCl} 3 -d) δ 157.2 (C-2 ', 153.6 (C-4''), 150.8 (C-2''), 150.7 (C-3), 142.5 (C-1 ''', 142.4 (C-3''), 132.0 (C-9'), 131.7 (C-4 ', 129.3 ', 129.0 (C-10', 126.8 (C-7 ', 125.9 (C-1''), 124.6 C-6 ''), 119.2 (C-3 ', 114.4 (C-2'''_{'-OCH 3), 60.9 (2} ''- OCH 3), 58.8 (C-5), 56.0 (4''- OCH 3), 47.6 (C-4); HREIMS (m / z):. Calcd for _{C 28 H 25 ClN 2 O 4} (M) +: 488.9621; Found 488.1500.

Synthesis of 1- (1- (4-chlorophenyl) -5- (2,4,5-trimethoxyphenyl) -4,5-dihydro-1H- pyrazol-3- yl) naphthalen-

Color: Yellow; mp 168-170 [deg.] C; Yield 53%; ¹ H NMR (400 MHz, CHCl ₃ -d)? 12.12 (s, IH, 2'-OH), 7.96 J = 1.3, 8.0 Hz), 7.75 (d, 1H, H-4 'J = 8.9 Hz), 7.42 (ddd, (D, 1H, H-6 'J = 1.3, 6.9, 8.0 Hz), 7.27 = 9.0 Hz), 6.93 (d, 2H, H-2 '''''' H-6 '' J = 9.0 Hz), 6.68 3 ''), 5.56 (dd, 1H, H-5, J = 7.0, 11.5 Hz), 4.33 (dd, _{'-OCH 3), 3.88 (s} , 3H, 4''- OCH 3), 3.65 (s, 3H, 5''- OCH 3), 3.55 (dd, 1H, H-4, J = 7.0, 16.8 Hz ); ^{13 C NMR (100 MHz, CHCl} 3 -d) δ 157.3 (C-2 ', 151.3 (C-3), 150.6 (C-2''), 149.5 (C-4''), 143.6 (C-5 ''), 142.9 (C-1 ''', 132.1 (C-9', 131.9 (C-4 ' 129.1 (C-10 ', 126.9 (C-7', 124.9 (C-4 ''), 123.4 (C-8 '), 123.2 C-3 '), 114.7 (C-2''' / C-6 '''), 110.5 (C-6''), 109.5 (C-5), 56.7 ( 5 '' - OCH 3), 56.4 (2 '' - OCH 3), 56.3 (4 '' - OCH 3), 47.4 (C-4); HREIMS (m / z): _{. Calcd for C 28 H 25 ClN} 2 O 4 (M) +: 488.9621; Found 488.1506.

Synthesis of 1- (5- (4-methoxyphenyl) -4,5-dihydro-1H-pyrazol-3- yl) naphthalen-2-ol (5)

Color: Light brown; mp 108-110 [deg.] C; Yield 66%; ^{1 H NMR (400 MHz, DMSO} -d 6) δ 10.66 (bs, 1H, 2'-OH), 8.13 (d, 1H, H-8 'J = 8.6 Hz), 7.81 (d, 1H, H-5 J = 8.0 Hz), 7.78 (d, 1H, H-4 'J = 9.0 Hz), 7.46 (d, J = 1.0, 6.9, 8.6 Hz), 7.41 (d, 2H, H-2 '' H-6 '', J = 8.7 Hz), 7.30 (ddd, 1H, , 8.0 Hz), 7.21 (d, 1H, H-3 ', J = 9.0 Hz), 6.93 (d, 2H, H-3' H-5, J = 3.5, 10.4, 10.4 Hz), 3.75 (s, 3H, 4 '' - OCH 3), 3.54 (dd, 1H, H-4, J = 10.4, 16.1 Hz), 3.08 (dd, 1H, H-4, J = 10.4, 16.1); ^{13 C NMR (100 MHz, DMSO} -d 6) δ 158.4 (C-4 ''), 154.5 (C-2 '), 149.1 (C-3), 134.7 (C-1''), 132.5 (C- (C-5 '), 128.0 (C-10'), 127.9 (C-2 '' / C-6 ''), 125.4 , 124.2 (C-8 '), 122.7 (C-6'), 118.3 (C-3 '), 113.7 C-5), 55.0 (4 '' - OCH 3), 44.9 (C-4); HREIMS (m / z): Calcd. _{for C 20 H 18 N 2 O} 2 (M) +: 318.3691; Found 318.1366.

Synthesis of 1- (5- (2,3-dimethoxyphenyl) -4,5-dihydro-1H-pyrazol-3-yl) naphthalen-2-ol

Color: Ivory; mp 109-111 [deg.] C; Yield 60%; ^{1 H NMR (400 MHz, DMSO} -d 6) δ 10.65 (bs, 1H, 2'-OH), 8.13 (d, 1H, H-8 'J = 8.5 Hz), 7.80 (d, 1H, H-5 (D, 1H, J = 8.0 Hz), 7.78 (d, 1H, H-4 ', J = 9.0 Hz), 7.43 H-1, J = 3.6 Hz), 7.30 (ddd, 1H, H-6 ', J = 1.0, 6.9, 8.0 Hz), 7.20 (dd, 1H, H-6 '', J = 1.4, 8.0 Hz), 7.09 (t, 1H, H-5 '', J = 8.0, 8.0 Hz), 6.98 , J = 1.4, 8.0 Hz) , 5.12 (td, 1H, H-5, J = 3.6, 10.6, 10.6 Hz), 3.81 (s, 3H, 3 '' - OCH 3), 3.77 (s, 3H, 2 _{'' -OCH 3), 3.59 (} dd, 1H, H-4, J = 10.6, 16.2 Hz), 3.00 (dd, 1H, H-4, J = 10.6, 16.2 Hz); ^{13 C NMR (100 MHz, DMSO} -d 6) δ 154.4 (C-2 '), 152.2 (C-3''), 148.9 (C-3), 146.2 (C-2''), 136.3 (C- 1 ''), 132.5 (C-9 '), 129.8 (C-4'), 128.1 (C-5 '), 128.0 '), 123.9 (C-5''), 122.7 (C-6'), 118.6 (C-6 ''), 118.3 ''), 60.2 (2 '' - OCH ₃ ), 57.5 (C-5), 55.6 (3 '' - OCH ₃ ), 44.3 (C-4); HREIMS (m / z): Calcd. _{for C 21 H 20 N 2 O} 3 (M) +: 348.3951; Found 348.1470.

Synthesis of 1- (5- (2,4-dimethoxyphenyl) -4,5-dihydro-1H-pyrazol-3-yl) naphthalen-2-ol

Color: Ivory; mp 145-146 [deg.] C; Yield 82%; ^{1 H NMR (400 MHz, DMSO} -d 6) δ 10.70 (bs, 1H, 2'-OH), 8.11 (d, 1H, H-8 ', J = 8.5 Hz), 7.80 (d, 1H, H- (Ddd, 1H, H-7 ', J = 1.0, 6.9, 8.5 Hz), 7.42 (d, (Ddd, 1H, H-6 ', J = 1.0, 6.9, 8.0 Hz), 7.28 ), 7.20 (d, 1H, H-3 ', J = 9.0 Hz), 6.58 (d, 1H, H-3' = 2.3, 8.4 Hz), 5.02 (td, 1H, H-5, J = 3.0, 10.0, 10.0 Hz), 3.80 (s, 3H, 2 '' - OCH 3), 3.76 (s, 3H, 4 '' _{-OCH 3), 3.55 (dd,} 1H, H-4, J = 10.0, 16.2 Hz), 2.95 (dd, 1H, H-4, J = 10.0, 16.2 Hz); ^{13 C NMR (100 MHz, DMSO} -d 6) δ 159.7 (C-4 ''), 157.6 (C-2 ''), 154.5 (C-2 '), 149.2 (C-3), 132.4 (C- 9 '), 130.0 (C-4'), 128.1 (C-5 '), 128.0 (C-10'), 127.0 '), 122.7 (C-6'), 122.6 (C-1 ''), 118.3 '), 57.4 (C-5 ), 55.4 (2''- OCH 3), 55.2 (4''- OCH 3), 43.3 (C-4); HREIMS (m / z):. Calcd for C 21 H ₂₀ N ₂ O ₃ (M) ⁺ : 348.3951; Found 348.1471.

Synthesis of 1- (5- (3,5-dimethoxyphenyl) -4,5-dihydro-1H-pyrazol-3-yl) naphthalen-2-ol (8)

Color: Ivory; mp 112-114 [deg.] C; Yield 55%; ^{1 H NMR (400 MHz, DMSO} -d 6) δ10.61 (bs, 1H, 2'-OH), 8.13 (d, 1H, H-8 ', J = 8.5 Hz), 7.81 (d, 1H, H 1H), 7.43 (ddd, 1H, H, J = 8.0 Hz), 7.79 (d, 1H, H-4 ', J = 8.9 Hz) J = 1.0, 6.9, 8.5 Hz), 7.30 (ddd, 1H, H-6 ', J = 1.0, 6.9, 8.0 Hz), 7.21 ), 6.67 (d, 2H, H-2 '' / H-6 '', J = 2.2 Hz), 6.41 1H, H-5, J = 3.0, 10.0, 10.0 Hz), 3.75 (s, 6H, 3 '' - OCH 3/5 '' - OCH 3), 3.56 (dd, 1H, H-4, J = 10.0 , 16.2 Hz), 3.10 (dd, 1H, H-4, J = 10.0, 16.2 Hz); ^{13 C NMR (100 MHz, DMSO} -d 6) δ 160.5 (C-3 '' / C-5 ''), 154.5 (C-2 '), 148.9 (C-3), 145.6 (C-1'' ), 132.4 (C-9 ', 129.9 (C-4'), 128.1 (C-5 '), 128.0 (C-3 '), 112.4 (C-1'), 104.5 (C-2 '' / C-6 ''), 98.9 -5), 55.1 (3 '' - OCH 3/5 '' - OCH 3), 44.7 (C-4); HREIMS (m / z):. Calcd for C 21 H 20 N 2 O 3 (m) + : 348.3951; Found 348.1470.

Example  2: General procedure for the synthesis of derivatives 9-17

1'-Hydroxy-2'-acetonaphthone (1,864 mg, 10 mmol) and methoxy-substituted benzaldehyde (10 mmol) were dissolved in 85 mL of ethanol and the temperature of the reaction mixture was adjusted to 5 DEG C in a cooling bath. To the cooled reaction mixture was added a solution of 50% KOH (10 mL) and the mixture was stirred at ambient temperature for 40 h. The reaction mixture was acidified by the addition of 6 N HCl (30 mL) to form a precipitate and re-crystallized with ethanol. When acidification did not produce a precipitate, the aqueous layer was extracted twice with dichloromethane. The combined aqueous layer was dried them with MgSO _4. Filtration and distillation of the solvent gave a residue which was purified by flash chromatography (ethyl acetate: hexane = 1: 4) to give the corresponding benzochalcones (VI). Benzochalcones (VI, 1 mmol) and hydrazine (1.2 mmol) were dissolved in ethanol (30 mL) and the reaction mixture was refluxed for 10 h. The temperature of the reaction mixture was adjusted to 5 캜 in a cooling bath to form a precipitate. Filtration and recrystallization gave pure derivatives 8-17.

Synthesis of 2- (5- (2,3-dimethoxyphenyl) -4,5-dihydro-1H-pyrazol-3-yl) naphthalen-1-ol (9)

Color: Ivory; mp 135-37 [deg.] C; Yield 90%; ¹ H-NMR (400 MHz, DMSO-d ₆ )? 12.32 (s, 1H, 1'-OH), 8.25 (dd, 1H, H-1, J = 3.5 Hz), 7.54 (td, 1H, H-6 ', J = 2.0, 6.8, 6.8 Hz), 7.51 1H, H-5 '), 7.41 (d, 1H, H-3', J = 9.0 Hz) H-5, J = 3.5, 7.0 Hz), 7.05 (m, 1H, H-6 ''), 7.00 (dd, 10.6, 10.6), 3.82 (s , 3H, 3 '' - OCH 3), 3.80 (s, 3H, 2 '' - OCH 3), 3.73 (dd, 1H, H-4, J = 10.6, 16.7 Hz) , 3.04 (dd, IH, H-4, J = 10.6,16.7 Hz); ¹³ C NMR (100 MHz, DMSO-d ₆ )? 153.2 (C-1 '), 133.7 (C-10'), 127.5 (C-5 ', 127.0 (C-3 '), 124.0 (C-5'), 123.9 (C-9 '), 122.3 4 '), 112.0 (C- 4''), 110.0 (C-2'), 60.3 (2 '' - OCH 3), 56.4 (C-5), 55.7 (3 '' - OCH 3), 40.5 ( C-4); HREIMS (m / z):. Calcd for C 21 H 20 N 2 O 3 (M) +: 348.3951; Found 348.1471.

Synthesis of 2- (5- (2-methoxyphenyl) -4,5-dihydro-1H-pyrazol-3-yl) naphthalen-1-ol (10)

Color: Yellow; mp 131-133 [deg.] C; Yield 74%; ^{1 H NMR (400 MHz, DMSO} -d 6) δ12.33 (s, 1H, 1'-OH), 8.24 (m, 1H, H-8 ', 7.85 (m, 1H, H-5'), 7.73 (d, 1H, H-1, J = 3.0 Hz), 7.54 (td, 1H, H-6 ', J = 2.0, 6.8, 6.8 Hz), 7.51 1H, J = 8.7 Hz), 7.41 (d, 1H, H-3 ', J = J = 0.8, 8.2 Hz), 6.95 (td, 1H, H-5 '', J = = 0.8, 7.5, 7.5 Hz) , 5.10 (td, 1H, H-5, J = 3.0, 10.2, 10.2 Hz), 3.85 (s, 3H, 2 '' - OCH 3), 3.70 (dd, 1H, H -4, J = 10.2, 16.7 Hz ), 3.00 (dd, 1H, H-4, J = 10.2, 16.7 Hz); 13 C NMR (100 MHz, DMSO-d 6) δ156.6 (C-2 '' ), 153.2 (C-3), 153.1 (C-1 '), 133.7 (C-10', 129.8 (C-6 ', 126.3 (C-6'), 125.4 (C-7 '), 124.7 (C-3'), 123.9 -5 ''), 118.5 (C -4 '), 110.8 (C-3''), 110.1 (C-2'), 56.4 (C-5), 55.5 (2 '' - OCH 3), 39.5 ( C-4); HREIMS (m / z): Calcd for C 20 H 18 N 2 O 2 (m) +:. 318.3691; Found 318.1371.

Synthesis of 2- (5- (3-methoxyphenyl) -4,5-dihydro-1H-pyrazol-3- yl) naphthalen-1-ol (11)

Color: Yellow; mp 118-120 [deg.] C; Yield 78%; ¹ H NMR (400 MHz, DMSO-d ₆ )? 12.28 (s, 1H, 1'-OH), 8.25 ), 7.86 (m, 1H, H-5 ', 7.54 (td, 1H, H-6' J = 1.9, 6.8, 6.8 Hz) H-4 ', 7.29 (t, 1H, H-5'', J = 8.1,8.1 Hz), 7.02 (d, 1H, H-2 ''), J = 1.6 Hz), 7.01 (m, 1H, H-6 ''), 6.87 , J = 3.6, 10.7, 10.7 Hz), 3.76 (s, 3H, 3 '' - OCH 3), 3.73 (dd, 1H, H-4, J = 10.7, 16.6 Hz), 3.10 (dd, 1H, H J = 10.7, 16.6 Hz); ¹³ C NMR (100 MHz, DMSO-d ₆ )? 159.3 (C-3 ''), 153.3 (C-10 '), 129.6 (C-5'), 127.5 (C-5 '), 127.1 (C-3 '), 123.9 (C-9'), 122.3 (C-8 '), 118.9 (C-2 ''), 110.0 (C-2 '), 62.1 (C-5), 55.0 (3''- OCH 3), 41.0 (C-4); HREIMS (m / z):. Calcd for _{C 20 H 18 N 2 O 2} (M) +: 318.3691; Found 318.1365.

Synthesis of 2- (5- (4-methoxyphenyl) -4,5-dihydro-1H-pyrazol-3- yl) naphthalen-1-ol (12)

Color: Yellow; mp 162 [deg.] C; Yield 96%; ^{1 H NMR (400 MHz, DMSO} -d 6) δ 12.31 (s, 1H, 1'-OH), 8.25 (m, 1H, H-8 ', 7.85 (m, 1H, H-5', 7.84 (d 1H, H-1 ', J = 3.4 Hz), 7.54 (td, 1H, H-6', J = 1.9, 6.8, 6.8), 7.52 6.8 Hz), 7.43 (s, 1H, H-3 ', 7.43 (s, 1H, H-4', 7.36 J = 8.7 Hz), 4.86 (td, 1H, H-5, J = 3.4, 10.7, 10.7 Hz), 3.75 (s, 3H _{, 4 '' - OCH 3)} , 3.68 (dd, 1H, H-4, J = 10.7, 16.6 Hz), 3.07 (dd, 1H, H-4, J = 10.7, 16.6 Hz); 13 C NMR (100 _{MHz, DMSO-d 6) δ} 158.6 (C-4 ''), 153.3 (C-3), 153.2 (C-1 '), 134.0 (C-1''), 133.7 (C-10'), 127.9 (C-6 '), 127.5 (C-5'), 127.0 (C-6 '), 125.5 C-5 '), 110.0 (C-2'), 61.7 (C-5 '), _{55.1 (4 '' - OCH 3} ), 41.0 (C-4); HREIMS (m / z):. Calcd for C 20 H 18 N 2 O 2 (m) +: 318.3691; Found 318.1366.

Synthesis of 2- (5- (2,4-dimethoxyphenyl) -4,5-dihydro-1H-pyrazol-3-yl) naphthalen-

Color: Ivory; mp 106-107 [deg.] C; Yield 82%; ^{1 H NMR (400 MHz, DMSO} -d 6) δ 12.34 (s, 1H, 1'-OH), 8.23 (m, 1H, H-8 '), 7.85 (m, 1H, H-5'), 7.65 (d, 1H, H-1, J = 3.2 Hz), 7.53 (td, 1H, H-6 ', J = 2.0, 6.8, 6.8 Hz), 7.51 , 6.8, 6.8 Hz), 7.43 (d, IH, H-4 ', J = 8.8 Hz), 7.41 (Dd, 1H, H-5 ', J = 2.4, 8.4 Hz), 5.02 (t, , 1H, H-5, J = 3.2, 10.2, 10.2 Hz), 3.83 (s, 3H, 2 '' - OCH 3), 3.75 (s, 3H, 4 '' - OCH 3), 3.63 (dd, 1H , H-4, J = 10.2, 16.6 Hz), 2.99 (dd, 1H, H-4, J = 10.2, 16.6 Hz); ^{13 C NMR (100 MHz, DMSO} -d 6) δ 159.8 (C-4 ''), 157.6 (C-2 ''), 153.3 (C-3), 153.1 (C-1 '), 133.6 (C- (C-6 '), 125.4 (C-7'), 124.7 (C-3 '), 123.9 (C-9' ), 122.3 (C-8 '), 122.0 (C-1''), 118.4 (C-4'), 110.1 '), 56.2 (C-5 ), 55.5 (2''- OCH 3), 55.2 (4''- OCH 3), 39.5 (C-4); HREIMS (m / z):. Calcd for C 21 H ₂₀ N ₂ O ₃ (M) ⁺ : 348.3951; Found 348.1470.

Synthesis of 2- (5- (3,5-dimethoxyphenyl) -4,5-dihydro-1H-pyrazol-3-yl) naphthalen-1-ol (14)

Color: Ivory; mp 107-111 [deg.] C; Yield 77%; ¹ H NMR (400 MHz, DMSO-d ₆ )? 12.26 (s, 1H, 1'-OH), 8.25 J = 1.9, 6.8, 6.8 Hz), 7.52 (td, 1H, H-7 ', J = 1.9 , 6.8, 6.8 Hz), 7.44 (d, IH, H-4 ', J = 8.8 Hz), 7.41 J = 2.3 Hz), 6.43 (d, 1H, H-4 '', J = 2.3 Hz), 4.85 (td, 1H, H-5, J = 3.6, 10.8, 10.8 Hz), 3.75 (s, 6H , 3 '' - OCH 3/5 '' - OCH 3), 3.69 (dd, 1H, H-4, J = 10.8, 16.6 Hz), 3.10 (dd, 1H, H- 4, J = 10.8, 16.6 Hz); ^{13 C NMR (100 MHz, DMSO} -d 6) δ 160.5 (C-3 '' / C-5 ''), 153.4 (C-3), 153.2 (C-1 '), 144.6 (C-1'' ), 123.7 (C-10 '), 127.5 (C-5'), 127.1 (C-6 '), 125.5 C-2 '), 104.7 (C-2''/C-6''), 99.0 (C-4''), 62.2 C-5), 55.1 (3 '' - OCH 3/5 '' - OCH 3), 41.0 (C-4); HREIMS (m / z): Calcd. _{for C 21 H 20 N 2 O} 3 (M) +: 348.3951; Found 348.1470.

Synthesis of 2- (5- (2,3,4-trimethoxyphenyl) -4,5-dihydro-1H-pyrazol-3- yl) naphthalen-1-ol (15)

Color: Yellow; mp 171-173 [deg.] C; Yield 96%; ¹ H-NMR (400 MHz, DMSO-d ₆ )? 12.34 (s, 1H, 1'-OH), 8.25 (d, 1H, H-1, J = 2.5 Hz), 7.54 (td, 1H, H-6 ', J = 1.9, 6.8, 6.8 Hz), 7.51 , 6.8, 6.8 Hz), 7.44 (d, IH, H-4 ', J = 8.8 Hz), 7.42 J = 8.7 Hz), 6.80 (d, 1H, H-5 ', J = 8.7 Hz), 5.02 (td, 1H, H-5, J = 2.5, 10.5, , 3H, 2 '' - OCH 3), 3.78 (s, 3H, 3 '' - OCH 3), 3.77 (s, 3H, 4 '' - OCH 3), 3.69 (dd, 1H, H-4, J = 10.5, 16.6 Hz), 3.03 (dd, 1H, H-4, J = 10.5, 16.6 Hz); ^{13 C NMR (100 MHz, DMSO} -d 6) δ 153.3 (C-3), 153.2 (C-1 '), 152.7 (C-3''), 151.1 (C-2''), 141.7 (C- 4 ''), 133.7 (C-10 '), 127.8 (C-1''), 127.5 3 '), 123.9 (C-9'), 122.3 (C-8 '), 121.3 (C-6'''), 60.9 (2 '' - OCH ₃ ), 60.3 (4 '' - OCH ₃ ), 56.5 (C-5), 55.8 (3 '' - OCH ₃ ), 40.4 (C-4); HREIMS (m / z): Calcd. _{for C 22 H 22 N 2 O} 4 (M) +: 378.4211; Found 378.1584.

Synthesis of 2- (5- (2,4,5-trimethoxyphenyl) -4,5-dihydro-1H-pyrazol-3- yl) naphthalen-

Color: Ivory; mp 140-142 [deg.] C; Yield 84%; ^{1 H NMR (400 MHz, DMSO} -d 6) δ 12.34 (s, 1H, 1'-OH), 8.24 (m, 1H, H-8 '), 7.86 (m, 1H, H-5', 7.69 ( J = 1.9, 6.8, 6.8 Hz), 7.51 (td, 1H, H-7 ', J = , 6.8, 6.8 Hz), 7.43 (d, 1H, H-4 ', J = 8.9 Hz), 7.41 ''), 6.74 (s, 1H, H-3 ''), 5.05 (td, 1H, H-5, J = 3.7, 10.8, 10.8 Hz), 3.83 (s, 3H, 2 '' - OCH 3) , 3.80 (s, 3H, 4 '' - OCH 3), 3.69 (s, 3H, 5 '' - OCH 3), 3.65 (dd, 1H, H-4, J = 10.8, 16.6 Hz), 2.98 (dd , ≪ / RTI > 1H, H-4, J = 10.8, 16.6 Hz); ^{13 C NMR (100 MHz, DMSO} -d 6) δ 153.5 (C-3), 153.2 (C-1 '), 151.1 (C-2''), 148.8 (C-4''), 142.5 (C- (C-10), 127.5 (C-5 '), 127.0 (C-6'), 125.4 '), 122.3 (C-8'), 121.0 (C-1 ''), 118.4 ''), 56.3 (2 '' - OCH ₃ ), 56.3 (5 '' - OCH ₃ ), 56.2 (C-5), 55.8 (4 '' - OCH ₃ ), 39.8 (C-4); HREIMS (m / z): Calcd. _{for C 22 H 22 N 2 O} 4 (M) +: 378.4211; Found 378.1579.

Synthesis of 2- (5- (2,4,6-trimethoxyphenyl) -4,5-dihydro-1H-pyrazol-3- yl) naphthalen-1-ol (17)

Color: Ivory; mp 160-162 [deg.] C; Yield 84%; ^{1 H NMR (400 MHz, DMSO} -d 6) δ 12.59 (s, 1H, 1'-OH), 8.24 (m, 1H, H-8 ', 7.85 (m, 1H, H-5', 7.53 (td , 7.42 (d, 1H, H-4 ', J'), 7.50 (td, 1H, H-7 ', J = 2.0, 6.8, 6.8 Hz) = 8.6 Hz), 7.38 (d, 1H, H-3 ', J = 8.6 Hz), 7.11 (bs, 1H, H- ), 5.35 (dd, 1H, H-5, J = 8.3, 12.6 Hz), 3.77 (s, 3H, 4 '' - OCH 3), 3.66 (s, 6H, 2 '' - OCH 3/6 '' _{-OCH 3), 3.41 (dd,} 1H, H-4, J = 12.6, 16.5 Hz), 3.30 (dd, 1H, H-4, J = 8.3, 16.5 Hz); 13 C NMR (100 MHz, DMSO- d ₆ )? 160.3 (C-4 ''), 159.2 (C-2 '' / C-6 ''), 153.4 , 127.4 (C-5 '), 126.7 (C-6'), 125.3 (C-7 '), 124.7 (C-4 '), 110.4 (C-1''), 110.3 (C-2'), 91.3 (C-3 '' / C-5 ''), 55.8 (2 '' - OCH 3/6 '_{'-OCH 3), 55.2 (4} ''- OCH 3), 50.5 (C-5), 38.2 (C-4); HREIMS (m / z):. Calcd for C 22 H 22 N 2 O 4 (M ) ^& Lt; ^{+ &} gt ^; : 378.4211; Found 378.1577.

Example  3: Biological essay

Colony formation essay

HCT116 human colon cancer cells were purchased from the American Type Culture Collection (Rockville, Md.). Cells were grown in Dulbecco's modified Eagle's medium (DMEM; Invitrogen Life Technologies, Carlsbad, Calif.) Supplemented with 10% (v / v) heat-inactivated fetal bovine serum (FBS; HyClone, Logan, UT). HCT116 cells (5 x 10 ³ cells / well) were seeded on 24-well tissue culture plates (BD Falcon, Franklin Lakes, NJ) in the presence or absence of 17 different derivatives of different concentrations and cultured for 7 days. HCT116 cell lines were tested on four types of plates: untreated plates and plates treated with 10, 20, and 40 [mu] M samples. The formed colonies were fixed with 6% glutaraldehyde and stained with 0.1% crystal violet [NA Franken, HM Rodermond, J. Stap, J. Haveman, C. van Bree, Nat. Protoc. 1 (2006) 2315-2319. Supplementary results of the colony formation survival assay at three different concentrations (10, 20, and 40 [mu] M) are shown in FIG. The sum of the three inhibition values measured using a densitometric assay was used as the biological date for each benzochalcone derivative. The lower values obtained from the densitometric analysis indicate the inhibition of HCT116 colon cancer cells.

Cell proliferation essay

HCT116 cells were seeded on 96-well plates (2 × 10 ³ cells / well) and treated with various concentrations (5-40 μM) of derivative 7 for 24 and 48 hours. Cell Counting 2- (2-methoxy-4-nitrophenyl) -3- (4-nitrophenyl) -5- (2,4-dihydroxyphenyl) -disulfophenyl) -2H-tetrazolium, monosodium salt]. The absorbance was read at 540 nm using a microplate reader (Molecular Devices Corp., Menlo Park, Calif.).

Western blot analysis

HCT116 cells were treated with 40 [mu] M of derivative 7 for 12, 24, or 48 h. Cells were disrupted in 20 mM HEPES (pH 7.2) containing 1% Triton X-100, 10% glycerol, 150 mM NaCl, 10 ug / mL leupeptin, and 1 mM phenylmethylsulfonyl fluoride (PMSF). The protein samples (15 μg each) were separated by 10% SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) and transferred to a nitrocellulose filter. The filter was blocked with 5% defatted milk powder and probed with an over-night primary antibody at 4 ° C and incubated for 2 hours at room temperature with the appropriate horseradish peroxidase-conjugated secondary antibody. Positive reactions were visualized with an enhanced chemiluminescence Western blotting detection system (Amersham Bioscience, Pascataway, NJ). Relative band intensities were measured with a quantitative scanning densitometer and image analysis software, Bio-1D version 97.04.

Flow cell analysis of cell cycle

HCT116 cells were treated with 40 μM of derivative 7 for 12, 24, or 48 h, collected, fixed with 70% ethanol, washed twice with PBS, and resuspended in 0.1% Triton X-100, 0.1 mM EDTA, / mL RNase A (50 ㎍ / mL propidium iodide (PI) solution). The amount of cellular DNA was analyzed by measuring fluorescence with a FACSCalibur Flow Cytometer (Becton Dickinson Immunocytometry Systems, San Jose, Calif.).

Claims (6)

1- (5- (2,4-Dimethoxyphenyl-4,5-dihydro-1H-pyrazol-3-yl) naphthalen-2-ol (ol). An anticancer pharmaceutical composition comprising the compound of claim 1 as an active ingredient. 3. The anticancer pharmaceutical composition according to claim 2, wherein the composition has an anticancer effect on colon cancer. delete delete Hydroxy-1'-acetonaphthone and 2,4-dimethoxy substituted benzaldehyde were dissolved in ethanol to react, and a base solution was added to the reaction mixture to prepare a benzalkonium derivative. Lt; RTI ID = 0.0 > 1 < / RTI > with a hydrazine.

Images