J. Chil. Chem. Soc., 57, N° 2 (2012), págs: 1118-1121.

 

NEW APPROACHES FOR THE SYNTHESIS OF HYDRAZONE DERIVATIVES AND THEIR ANTITUMOR EVALUATION

 

WAGNAT WAHBA WARDAKHAN* AND EMAN MOHAMED SAMIR

National Organization for Drug Control & Research (NODCAR) P.O. 29, Cairo, A. R. Egypt. e-mail: wagnatward@hotmail.com

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ABSTRACT

The hydrazide-hydrazone 3 reacted with benzenediazonium chloride to give the phenylhydrazone derivative 5. The latter underwent a series of hetero-cyclization reactions to give pyridazine, 1,2,3-triazole and pyrazole derivatives. The antitumor evaluation of the newly synthesized products against the three cancer cells lines namely breast adenocarcinoma (MCF-7), non-small cell lung cancer (NCI-H460) and CNS cancer (SF-268) showed that some of them have higher inhibitory effects towards the three cell lines compared to the standard.

Key word: hydrazide-hydrazone, pyridazine, 1,2,3-triazole, pyrazole, anti-tumor.

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INTRODUCTION

Hydrazide-/hydrazones have been demonstrated to possess antibacterial,^1-7 anticonvulsant^8-11 and antitubercular^12-16 activities. These observations led us to synthesize novel hydrazide-hydrazones and to investigate their possible antitumor activities. It has been reported in the literature^17,18 that hydrazide-hydrazones can give the corresponding hydrazide and aldehyde metabolites whereas the related hydrazides are known to yield carboxylic acids via hydrolytic route. Based on this knowledge, hydrazide-hydrazones derivative 3 was prepared easily from the reaction of cyanoacetyl hydrazine with benzaldehyde and subsequently used for the synthesis of pyridazine, 1,2,3-triazole and pyrazole derivatives. The newly synthesized compounds were tested to evaluate their in vitro anti-tumor activities against three human tumor cell lines representing different tumor types, namely, breast adenocarcinoma (MCF-7), non-small cell lung cancer (NCI-H460) and CNS cancer (SF-268). It was found that some of these compounds showed inhibitory effects on the three cell lines, indicating their potential use in the development of anti-cancer agents.

RESULTS AND DISCUSION

The reaction of the cyanoacetylhydrazine 1 with benzaldehyde 2 gives the hydrazide-hydrazone 3.¹⁹ Reaction of the latter product with benzenediazonium chloride 4 afforded the phenylhydrazone derivative 5. The structure of compound 5 was confirmed on the basis of analytical and spectral data. Heterocyclization of compound 5 to form different heterocyclic derivatives with potential pharmaceutical applications was studied. Thus, compound 5 reacted with either malononitrile 6a or ethyl cyanoacetate 6b in 1,4-dioxan containing a catalytic amount of triethylamine, gave the pyridazine derivatives 7a and 7b, respectively. The structures of compounds 7a & 7b were established on the basis of their analytical and spectral data. Thus, the ¹H NMR spectrum of 7a showed the presence of a singlet at δ 4.87 ppm corresponding to NH₂ group, a singlet at δ 6.69 ppm for the CH=N group, a multiplet at δ 7.28-7.58 ppm for the two phenyl groups and two singlets (D₂O exchangeable) at δ 8.23 and 8.30 ppm for the two NH groups. On the other hand, reaction of compound 5 with hydrazine hydrate gave the triazole derivative 9. Moreover, the reaction of compound 5 with phenylisothiocyanate 10 gave the 1,2,4-triazine derivative 12. The structures of compounds 9 and 12 were established on the basis of their analytical and spectral data (see experimental section).

Next, the reaction of compound 3 with phenylisothiocyanate in DMF/ KOH solution was studied. This reaction led to the formation of the nonisolable intermediate potassium sulphide salt 13 which subsequently allowed to react with hydrazine hydrate to afford the hydrazino derivative 14 was formed. The structure of the latter product was confirmed on the basis of its analytical and spectral data. Thus, the ¹H NMR spectrum showed the presence of a singlet at δ 5.02 ppm corresponding to NH₂ group, a singlet at δ 6.99 for the CH=N group, a multiplet at δ 7.28-7.49 ppm corresponding to the two phenyl group and three singlets at δ 8.02, 8.90, 9.21 ppm corresponding to the three NH groups. Compound 14 was cyclized readily in sodium ethoxide solution affording the 3-amino-5-(phenylamino)-1H-pyrazole-4-N-benzalcarbohydrazide derivative 15. The structure of the latter product was established based on its analytical and spectral data (see experimental sections). Diazotization of the 3-amino group of compound 15 using sodium nitrite solution in the presence of acetic/ hydrochloric acids resulted in the formation of intermediate pyrazole-3-diazonium salt 16. Coupling of the latter diazonium salt with malononitrile in ethanol containing sodium acetate afforded the hydrazo derivative 17. The analytical and spectral data of the obtained product were consistence with the proposed structure (Scheme 2).

The reaction of compound 14 with either acetylacetone 18a or ethyl acetoacetate 18b gave the pyrazole derivatives 19a and 19b, respectively. In a similar manner, the reaction of 14 with either malononitrile 6a or ethyl cyanoacetate 6b gave the pyrazole derivatives 20a and 20b, respectively. Moreover, the reaction of 14 with either a-cyanocinnamonitrile 21a or ethyl a-cyanocinnamate 21b gave the 5-phenylpyrazole derivatives 22a and 22b, respectively (Scheme 3). The analytical and spectral data of compounds 19a,b-22a,b were in agreement with their structures (see experimental section).

Material, methods & Reagents: Fetal bovine serum (FBS) and L-glutamine, were obtained from Gibco Invitrogen Co. (Scotland, UK). RPMI-1640 medium was from Cambrex (New Jersey, USA). Dimethyl sulfoxide (DMSO), doxorubicin, penicillin, streptomycin and sulforhodamine B (SRB) from Sigma Chemical Co. (Saint Louis, USA). Samples: Stock solutions of compounds 5-22b were prepared in DMSO and kept at -20 ^oC. Appropriate dilutions of the compounds were freshly prepared just prior the assays. Final concentrations of DMSO did not interfere with the cell growth.

Cell cultures: Three human tumor cell lines, MCF-7 (breast adenocarcinoma), NCI-H460 (non-small cell lung cancer), and SF-268 (CNS cancer) were used. MCF-7 was obtained from the European Collection of Cell Cultures (ECACC, Salisbury, UK) and NCI-H460 and SF-268 were kindly provided by the National Cancer Institute (NCI, Cairo, Egypt). They grow as monolayer and routinely maintained in RPMI-1640 medium supplemented with 5% heat inactivated FBS, 2 mM glutamine and antibiotics (penicillin 100 U/mL, streptomycin 100 mg/mL), at 37 ^oC in a humidified atmosphere containing 5% CO₂. Exponentially growing cells were obtained by plating 1.5 X 10⁵ cells/mL for MCF-7 and SF-268 and 0.75 X 10⁴ cells/mL for NCI-H460, followed by 24 h of incubation. The effect of the vehicle solvent (DMSO) on the growth of these cell lines was evaluated in all the experiments by exposing untreated control cells to the maximum concentration (0.5%) of DMSO used in each assay.

Results are given in concentrations that were able to cause 50 % of cell growth inhibition (GI₅₀) after a continuous exposure of 48 h and show means ± SEM of three-independent experiments performed in duplicate.

Effect on the growth of human tumor cell lines

The effect of compounds 5-22b was evaluated on the in vitro growth of three human tumor cell lines representing different tumor types, namely, breast adenocarcinoma (MCF-7), non-small cell lung cancer (NCI-H460) and CNS cancer (SF-268), after a continuous exposure of 48 h. The results are summarized in Table 1.

All the compounds were able to inhibit the growth of the human tumor cell lines in a dose-dependent manner (data not shown). Compounds 5 and 20b showed the best results, exhibiting an equivalent potency in all the three tumor cell lines which is still much lower than the control doxorubicin, On the other hand, compounds 7a, 14, 19a and 20a showed moderated growth inhibitory effect. Comparing the activities of 20a and 20b it is observed that OH group present in 20b presents a stronger growth inhibitory effect than compound 20a with the NH₂ group. It is obvious that compounds 7b, 15, 17, 19b, 22a showed the lowest inhibitory effect towards the three cell lines.

EXPERIMENTAL SECTION

All melting points were determined in open capillaries and are uncorrected. IR spectra were measured using KBr discs on a Pye Unicam SP-1000 spectrophotometer. ¹H-NMR spectra were measured on a Varian EM390-200 MHz instrument in CD₃SOCD₃ as solvent using TMS as internal standard, and chemical shifts are expressed as δ in units of parts per million (ppm). Analytical data were obtained from the Micro analytical data unit at Cairo University.

2- Cyano-2-phenylhydrazo-N-benzalacetylhydrazide (5):

To a cold solution (0-5 °C) of compound 3 (1.87 g, 0.01 mol) in ethanol (50 mL) containing sodium acetate (4.0 g), benzenediazonium chloride [prepared by adding sodium nitrite solution (0.70 g in 10 mL water) to a cold solution (0-5 ^°C) of aniline (0.94 g, 0.01 mol) in concentrated hydrochloric acid (10 mL) with continuous stirring] was added with continuous stirring. The reaction mixture was then stirred at room temperature for additional 1 h and the formed solid product was collected by filtration.

Orange crystals from ethanol, yield (2.03 g, 70 %), m.p. 70 °C . IR (KBr): υ/cm^-1 = 3522-3332 (2NH), 3057 (CH aromatic), 2222 (CN), 1690 (CO), 1645 (C=C), 1637 (C=N). ¹H NMR (DMSO) = 6.56 (s, 1H, CH=N), 7.30-7.39 (m, 10H, 2C₆H₅), 8.30, 8.37 (2s, 2H, D₂O exchangeable, 2NH). Calcd for C₁₆H₁₃N₅O (291.31): C, 65.97; H, 4.50; N, 24.04 %. Found: C, 66.01; H, 4.48; N, 24.33 %. MS m/z: 291 (M⁺).

5-Amino-4-cyano-3-imino-1-phenylpyridazin-6-N-benzalcarbohydra-zide (7a) and 5-Amino-4-cyano-3-oxo-1-phenylpyridazin-6-N-benzalcar-bohydrazide (7b)

General procedure: To a solution of compound 5 (1.91 g, 0.01 mol) in 1,4-dioxan (40 mL) containing triethylamine (0.50 mL), either malononitrile (0.66 g, 0.01 mol) or ethyl cyanoacetate (1.25 g, 1.18 mL, 0.01 mol) was added. The reaction mixture was heated under reflux for 3 h then poured onto ice/water mixture containing few drops of hydrochloric acid and the formed solid product was collected by filtration.

Compound 7a: Yellow crystals from 1,4-dioxan, yield (2.74 g, 77 %), m.p. 90 °C. IR (KBr): υ/cm^-1 = 3480-3330 (NH₂, 2NH), 3060 (CH aromatic), 2223 (CN), 1688 (CO), 1638 (C=C), 1628 (C=N). ¹HNMR (DMSO) δ = 4.87 (s, 2H, D₂O exchangeable, NH₂), 6.69 (s, 1H, CH=N), 7.28-7.58 (m, 10H, 2C₆H₅), 8.23, 8.30 (2s, 2H, D₂O exchangeable, 2NH). Calcd for C₁₉H₁₅N₇O (357.37): C, 63.86; H, 4.23; N, 27.44 %. Found: C, 64.08; H, 4.47; N, 27.72 %. MS m/z: 357 (M⁺).

Compound 7b: Pale orange crystals from 1,4-dioxan, yield (2.74 g, 77 %), m.p.160 °C. IR (KBr): υ/cm^-1 = 3477-3328 (NH₂, NH), 3055 (CH aromatic), 2220 (CN), 1686 (2 CO), 1636 (C=C), 1625 (C=N). ¹H NMR (DMSO) δ = 4.85 (s, 2H, D₂O exchangeable, NH₂), 6.66 (s, 1H, CH=N), 7.29-7.49 (m, 10H, 2C₆H₅), 8.26 (s, 1H, D₂O exchangeable, NH). Calcd for C₁₉H₁₄N₆O₂ (358.35): C, 63.68; H, 3.94; N, 23.45 %. Found: C, 63.84; H, 4.02; N, 23.66 %. MS m/z: 358 (M⁺).

4- Amino-2-phenyl-5-N-benzalcarbohydrazido-1,2,3-triazole (9)

General procedure: To a solution of compound 5 (1.91 g, 0.01 mol) in 1,4-dioxan (40 mL), hydrazine hydrate (0.50 g, 0.01 mol) was added. The reaction mixture was heated under reflux for 4 h then poured onto ice/water containing few drops of hydrochloric acid. The formed solid product was collected by filtration.

Yellow crystals from 1,4-dioxan, yield (2.45 g, 80 %), m.p. 60 ^°C. IR (KBr): υ/cm^-1 = 3467-3338 (NH₂, NH), 3059 (CH aromatic), 1689 (CO), 1632 (C=C), 1620 (C=N). ¹H NMR (DMSO) δ = 4.89 (s, 2H, D₂O exchangeable, NH₂), 6.68 (s, 1H, CH=N), 7.20-7.36 (m, 10H, 2C₆H₅), 8.22 (s, 1H, D₂O exchangeable, NH). Calcd for C₁₆H₁₄N₆O (306.32): C, 62.74; H, 4.61; N, 27.44 %. Found: C, 62.44; H, 4.52; N, 27.71 %. MS m/z: 306 (M⁺).

5- Amino-2,4-diphenyl-3-thioxo-6-N-benzalcarbohydrazido-1,3,4-triazine (12)

To a solution of compound 5 (1.91 g, 0.01 mol) in 1,4-dioxan (40 mL), phenylisothiocyanate (1.35 g, 0.01 mol) was added. The reaction mixture was heated under reflux for 4 h then poured onto ice/water containing few drops of hydrochloric acid. The formed solid product was collected by filtration.

Yellow crystals from 1,4-dioxan, yield (3.42 g, 80 %), m.p. 40 ^°C. IR (KBr): υ/cm^-1 = 3438-3322 (NH₂, NH), 3052 (CH aromatic), 1685 (CO), 1630 (C=C), 1628 (C=N). ¹H NMR (DMS): δ = 4.68 (s, 2H, D₂O exchangeable, NH₂), 6.72 (s, 1H, CH=N), 7.28-7.42 (m, 16H, 3C₆H₅ pyridazine CH), 8.29 (s, 1H, D₂O exchangeable, NH). Calcd for C₂₃H₂₀N₆OS (428.51): C, 64.47; H, 4.70; N, 19.61; S, 7.48 %. Found: C, 64.55; H, 4.66; N, 19.55; S, 7.71 %. MS m/z: 428 (M⁺).

α -Cyano-b-phenylamino-β-hydrazinoylidino-N-benzalacrylohydrazi-de (14)

To a stirred solution of compound 5 (1.91 g, 0.01 mol) in dimethylfor-mamide (20 mL) containing potassium hydroxide (0.56 g, 0.01 mol) phenyli-sothiocyanate (1.35 g, 0.01 mol) was added. The whole reaction mixture was kept at room temperature overnight then hydrazine hydrate (0.50 g, 0.01 mol) was added. The formed solid product was collected by filtration.

White crystals from ethanol, yield (2.81 g, 88 %), m.p. 110 °C. IR (KBr): υ/cm^-1 = 3459-3331 (NH₂, 3NH), 3056 (CH aromatic), 1689 (CO), 1631 (C=C), 1623 (C=N). H¹NMR (DMSO) δ = 4.77 (s, 2H, D₂O exchangeable, NH₂), 6.59 (s, 1H, CH=N), 7.22-7.38 (m, 10H, 2C₆H₅), 8.24, 8.44, 8.50 (3s, 3H, D₂O exchangeable, 3NH). Calcd for C₁₇H₁₆N₆O (320.35): C, 63.74; H, 5.03; N, 26.23 %. Found: C, 64.01; H, 4.89; N, 26.51 %. MS m/z: 320 (M+).

3-Amino-5-phenylamino-3-N-benzalcarbohydrazido-pyrazole (15)

A suspension of compound 14 (3.20 g, 0.01 mol) in sodium ethoxide solution prepared by dissolving sodium metal (0.64 g, 0.02 mol) in absolute ethanol (40 mL) was heated in a boiling water bath for 3 h. The reaction mixture was left to cool and the formed solid product upon pouring onto ice/ water containing few hydrochloric acid (till pH 6) was collected by filtration.

Pale yellow crystals from ethanol, yield (2.21 g, 69 %), m.p. 60 ^°C. IR (KBr): υ/cm^-1 = 3446-3323 (NH₂, 3NH), 3051 (CH aromatic), 1686 (CO), 1630 (C=C), 1626 (C=N). ¹H NMR (DMSO) δ = 4.82 (s, 2H, NH₂), 6.61 (s, 1H, CH=N), 7.29-7.39 (m, 10H, 2C₆H₅), 8.20, 8.76, 8.82 (3s, 3H, D₂O exchangeable, 3NH). Calcd for C₁₇H₁₆N₆O (320.35): C, 63.74; H, 5.03; N, 26.23 %. Found: C, 63.71; H, 5.24; N, 26.49 %. MS m/z: 320 (M+).

3-Hydrazodicyanoylideno-5-phenylamino-3-N-benzalcarbohydrazi-dopyrazole (17)

A solution of the 3-diazo-5-phenylamino-3-N-benzalcarbohydrazidopyr-azole (0.01 mol) [prepared by adding sodium nitrite solution (0.70 g, 0.01 mol) to a cold solution (0 ^°C) of compound 15 (3.20 g, 0.01 mol) in acetic acid (20 mL) and concentrated hydrochloric acid (10 mL) with continuous stirring] was added to a cold solution of malononitrile (0.66 g, 0.01 mol) in ethanol (40 mL) containing sodium acetate (4.0 g) with continuous stirring for 2 h. The formed solid product was collected by filtration.

Orange crystals from ethanol, yield (3.02 g, 76 %), m.p. 160 °C. IR (KBr): υ/cm^-1 = 3560-3328 (4NH), 3046 (CH aromatic), 2227, 2220 (2CN), 1691 (CO), 1633 (C=C), 1626 (C=N). ¹H NMR (DMSO) δ = 6.70 (s, 1H, CH=N), 7.31-7.40 (m, 10H, 2C₆H₅), 8.14, 8.23, 8.40, 8.66 (4s, 4H, D₂O exchangeable, 4NH). Calcd for C₂₀H₁₅N₉O (397.39): C, 60.45; H, 3.80; N, 31.72 %. Found: C, 60.53; H, 4.48; N, 32.06 %. MS m/z: 397 (M+).

α-Cyano b-phenylamino-β-(3,5-dimethylpyrazol-1-yl)-N-benzalacrylo-hydrazide (19a) and α-Cyno-β-phenylamino-β-(3-hydroxy-5-methylpyrazol-1-yl)-N-benzalacrylohydrazide (19b)

General procedure: To a solution of compound 14 (3.20 g, 0.01 mol) in 1,4-dioxan (30 mL) either acetylacetone (1.0 g, 0.01 mol) or ethyl acetoacetate (1.30 g, 0.01 mol) was added. The reaction mixture was heated under reflux for 2 h then the solvent was evaporated under reduced pressure. The remaining residue was triturated with ethanol and the formed solid product was collected by filtration.

Compound 19a: Orange crystals from 1,4-dioxan, yield (2.76 g, 72 %), m.p. 70 °C. IR (KBr): υ/cm^-1 = 3487-3335 (2NH), 3055 (CH aromatic), 2225 (CN), 1685 (CO), 1630 (C=C), 1623 (C=N). ¹H NMR (DMSO) δ = 2.85, 3.03 (2s, 6H, 2CH₃), 5.99 (s, 1H, pyrazole H-4), 6.73 (s, 1H, CH=N), 7.29-7.37 (m, 10H, 2C₆H₅), 8.19, 8.25 (2s, 2H, D₂O exchangeable, 2NH). Calcd for C₂₂H₂₀N₆O (384.43): C, 68.73; H, 5.24; N, 21.86 %. Found: C, 68.52; H, 5.46; N, 22.04%. MS m/z: 384 (M⁺).

Compound 19b: Pale yellow crystals from 1,4-dioxan, yield (2.56 g, 66 %), m.p.100 °C. IR (KBr): υ/cm^-1 = 3592-3330 (OH, 2NH), 3058 (CH aromatic), 2223 (CN), 1688 (CO), 1633 (C=C), 1620 (C=N). ¹H NMR (DMSO) δ = 2.89 (s, 3H, CH₃), 5.97 (s, 1H, pyrazole H-4), 6.71 (s, 1H, CH=N), 7.26-7.39 (m, 10H, 2C₆H₅), 8.15, 8.22 (2s, 2H, D₂O exchangeable, 2NH), 10.22 (s, 1H, D₂O exchangeable, OH). Calcd for C₂₁H₁₈N₆O₂ (386.41): C, 65.27; H, 4.70; N, 21.75 %. Found: C, 65.31; H, 5.02; N, 22.01%. MS m/z: 386 (M+).

α -Cyano-b-phenylamino-β-(3,5-diaminopyrazol-1-yl)-N-benzalacrylo-hydrazide (20a) and α -Cyano-β-phenylamino-β-(3-hydroxy-5-aminopyrazol-1-yl)-N-benzalacrylohydrazide (20b)

General procedure: To a solution of compound 14 (3.20 g, 0.01 mol) in 1,4-dioxan (30 mL) either malononitrile (0.66 g, 0.01 mol) or ethyl cyanoacetate (1.25 g, 0.01 mol) was added. The reaction mixture was heated under reflux for 4 h then the solvent was evaporated under reduced pressure. The remaining residue was triturated with ethanol and the formed solid product was collected by filtration.

Compound 20a: Orange crystals from 1,4-dioxan, yield (2.76 g, 66 %), m.p. 60 °C. IR (KBr): υ/cm^-1 = 3487-3335 (2NH), 3058 (CH aromatic), 2228 ( CN), 1689 (CO), 1633 (C=C), 1622 (C=N). ¹H NMR (DMSO) δ = 4.66, 5.23 (2s, 4H, D₂O exchangeable, 2NH₂), 5.63 (s, 1H, pyrazole H-4), 6.80 (s, 1H, CH=N), 7.26-7.39 (m, 10H, 2C₆H₅), 8.20, 8.29 (2s, 2H, D₂O exchangeable, 2NH). Calcd for C₂₀H₁₈N₈O (386.41): C, 62.17; H, 4.70; N, 29.00 %. Found: C, 60.93; H, 5.83; N, 28.37%. MS m/z: 386 (M⁺).

Compound 20b: Buff crystals from ethanol, yield (2.56 g, 66 %), m.p. 70 °C. IR (KBr): υ/cm^-1 = 3569-3324 (OH, 2NH & NH₂), 3056 (CH aromatic), 2222 (CN), 1690 (CO), 1630 (C=C), 1622 (C=N). ¹H NMR (DMSO) δ = 4.66 (s, 2H, D₂O exchangeable, NH₂), 5.89 (s, 1H, pyrazole H-4), 6.69 (s, 1H, CH=N), 7.24-7.40 (m, 10H, 2C₆H₅), 8.18, 8.26 (2s, 2H, D₂O exchangeable, 2NH), 10.09 (s, 1H, D₂O exchangeable, OH). Calcd for C₂₀H₁₇N₇O₂ (387.39): C, 62.01; H, 4.42; N, 25.31 %. Found: C, 61.89; H, 4.63; N, 25.44%. MS m/z: 387 (M⁺).

α -Cyano-b-phenylamino-β-(3-amino-4-cyano-5-phenylpyrazol-1-yl)-N-benzalacrylohydrazide (22a) and α-Cyno-b-phenylamino-β-(3-hydroxy-3-cyano-5-phenylpyrazol-1-yl)-N-benzalacrylohydrazide (22b)

General procedure: To a solution of compound 14 (3.20 g, 0.01 mol) in 1,4-dioxan (30 mL) either a-cyanocinnamonitrile (1.54 g, 0.01 mol) or ethyl a-cyanocinnamate (2.01 g, 0.01 mol) was added. The reaction mixture was heated under reflux for 4 h then evaporated under vacuum. The remaining product was triturated with ethanol and the formed solid product was collected by filtration.

Compound 22a: Pale yellow crystals from 1,4-dioxan, yield (4.17 g, 88 %), m.p. 60 °C IR (KBr): υ/cm^-1 = 3463-3324 (2NH& NH₂), 3056 (CH aromatic), 2224 (CN), 1689 (CO), 1631 (C=C), 1620 (C=N). ¹H NMR (DMSO) δ = 4.69 (s, 2H, D₂O exchangeable, NH₂), 6.77 (s, 1H, CH=N), 7.307.42 (m, 15H, 3C₆H₅), 8.21, 8.32 (2s, 2H, D₂O exchangeable, 2NH). Calcd for C₂₇H₂₀N₈O (472.18): C, 68.63; H, 4.27; N, 23.72 %. Found: C, 68.41; H, 4.08; N, 23.68 %. MS m/z: 472 (M⁺).

Compound 22b : Pale brown crystals from ethanol, yield (3.83 g, 60 %), m.p.70 °C. IR (KBr): υ/cm^-1 = 3584-3348 (OH, 2NH), 3051 (CH aromatic), 2220 (CN), 1690 (CO), 1630 (C=C), 1622 (C=N). ¹H NMR (DMSO) δ = 6.59 (s, 1H, CH=N), 7.27-7.37 (m, 15H, 3C₆H₅), 8.22, 8.29 (2s, 2H, D₂O exchangeable, 2NH), 10.32 (s, 1H, D₂O exchangeable, OH). Calcd for C₂₇H₁₉N₇O₂ (473.49): C, 68.49; H, 4.04; N, 20.71 %. Found: C, 68.72; H, 4.28; N, 20.94%. MS m/z: 473 (M⁺).

CONCLUSIONS

The work described showed the uses of the phenylhydrazone derivative 5 in a series of heterocyclic transformations to form pyridazine, 1,2,4-triazine and pyrazole derivatives. The antitumor evaluation of the synthesized products showed that the pyrazole derivative 20b has the maximum inhibitory effect towards the cancer cell lines.

 

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(Received: August 2, 2011 - Accepted: January 23, 2012).