Molecules 2013, 18(3), 2683-2711; doi:10.3390/molecules18032683

Article
Synthesis, Antibacterial and Antifungal Activity of Some New Pyrazoline and Pyrazole Derivatives
Seham Y. Hassan
Department of Chemistry, Faculty of Science, University of Alexandria, PO Box 426, Ibrahimia 21321, Alexandria, Egypt; E-Mail: sehamyassen@yahoo.com; Tel.: +20-01-286-525-997; Fax: +20-3-5932-488
Received: 31 October 2012; in revised form: 7 January 2013 / Accepted: 9 January 2013 /
Published: 28 February 2013

Abstract

: A series of 2-pyrazolines 59 have been synthesized from α,β-unsaturated ketones 24. New 2-pyrazoline derivatives 1315 bearing benzenesulfonamide moieties were then synthesized by condensing the appropriate chalcones 24 with 4-hydrazinyl benzenesulfonamide hydrochloride. Ethyl [1,2,4] triazolo[3,4-c][1,2,4]triazino[5,6-b]-5H-indole-5-ethanoate (26) and 1-(5H-[1,2,4]triazino[5,6-b] indol-3-yl)-3-methyl-1H-pyrazol-5(4H)-one (32) were synthesized from 3-hydrazinyl-5H-[1,2,4]triazino[5,6-b]indole (24). On the other hand ethyl[1,2,4]triazolo[3,4-c][1,2,4]triazino[5,6-b]-5,10-dihydroquinoxaline-5-ethanoate (27) and 1-(5,10-dihydro-[1,2,4]triazino[5,6-b]quinoxalin-3-yl)-3-methyl-1H-pyrazol-5(4H)-one (33) were synthesized from 3-hydrazinyl-5,10-dihydro-[1,2,4]triazino[5,6-b]quinoxaline (25) by reaction with diethyl malonate or ethyl acetoacetate, respectively. Condensation of 6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indole-2-carbaldehyde (1') with compound 24 or 25 afforded the corresponding Schiff's bases 36 and 37, respectively. Reaction of the Schiff's base 37 with benzoyl hydrazine or acetic anhydride afforded benzohydrazide derivative 39 and the cyclized compound 40, respectively. Furthermore, the pyrazole derivatives 4244 were synthesized by cyclization of hydrazine derivative 25 with the prepared chalcones 24. All the newly synthesized compounds have been characterized on the basis of IR and 1H-NMR spectral data as well as physical data. Antimicrobial activity against the organisms E. coli ATCC8739 and P. aeruginosa ATCC 9027 as examples of Gram-negative bacteria, S. aureus ATCC 6583P as an example of Gram-positive bacteria and C. albicans ATCC 2091 as an example of a yeast-like fungus have been studied using the Nutrient Agar (NA) and Sabouraud Dextrose Agar (SDA) diffusion methods. The best performance was found for the compounds 16, 17, 19 and 20.
Keywords:
chalcones; pyrazoline; pyrazole; carbothioamide; benzenesulfonamide; thiazolidine; indole; quinoxaline

1. Introduction

Chalcones have been recently the subject of great interest due to their interesting pharmacological activities, including antioxidant [1,2], antibacterial [3], antileishmanial [4], anticancer [5], antiangiogenic [6], anti-infective, anti-inflammatory [7], antifungal [8], anti-malarial [9], anti-tumor [10], anti-protozoal [11] and cytotoxic properties [12]. Many pyrazole derivatives are reported to have a broad spectrum of biological activities, such as anti-inflammatory [13], antifungal [14], antiviral [15], cytotoxic [12], A3 adenosine receptor antagonists [16], antioxidant [13], antihypertensive [17], tranquilizing, muscle relaxant, psychoanaleptic, hypnotic, ulcerogenic, antidepressant, antibacterial and analgesic effects [18]. Pharmacologically-interesting heterocyclic systems like pyrazolines have been widely studied owing to their pharmacological activities, which include anti-tumor [19,20], anti-inflammatory [21,22,23,24,25,26,27,28,29,30,31,32], anti-parasitary [33], anticonvulsant [34], antimicrobial [35,36,37,38,39], antinociceptives [40], antimalarial [41], nitric oxide synthase inhibitory, associated with diseases such as Alzheimer, Huntington, and inflammatory arthritis [42], antidepressant [43,44], anticancer [45,46,47], antibacterial [48], antitubercular, analgesic [49], antiviral [46], antioxidant [50], antiamoebic [51,52,53], cytotoxic [53], antidiabetic [20], antifungal [54,55], antinociceptive [56], antimycobacterial [57], antihepatotoxic [58] and pesticidal properties [59].

Substituted 2-pyrazolines have been synthesized from α,β-unsaturated ketones and hydrazine hydrate with acetic/formic acid in ethanol/dimethyl sulfoxide (DMSO) [60], hydrazine in dimethyl formamide (DMF) or acetic acid [46], nicotinic acid hydrazide in n-butanol [41], phenyl hydrazine hydrochloride in the presence of sodium acetate [39], hydrazine hydrate in ethanol and DMF [25], and phenyl hydrazine in the presence of hot pyridine [27]. Some new substituted 2-pyrazoline derivatives bearing benzenesulfonamide moieties [21,22,23,26] were synthesized by condensing appropriate chalcones with 4-hydrazinobenzenesulfonamide hydrochloride. In view of these observations and in continuation of our research programme on the synthesis of five-membered heterocyclic compounds [61,62,63,64,65,66], we report herein the synthesis of some new pyrazoline and pyrazole derivatives bearing an indoline and quinoxaline moiety, which have been found to possess an interesting profile of antimicrobial activity.

2. Results and Discussion

2.1. Chemistry

2.1.1. Preparation of the Chalcones 2–4

The chalcones 24 were prepared as starting material to obtain the desired pyrazoline and pyrazole derivatives. The sequence leading to the title compounds is outlined in Scheme 1. The desired compounds were prepared by the reaction of 6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indole-2-carbaldehyde (1') [67] with different acetophenones (p-bromo-, p-chloro-, or p-methoxyacetophenones) in aqueous ethanolic KOH in good yield (Scheme 1). Their 1H-NMR spectra showed the -CH=CH- protons as a multiplet in the 7.52–7.63 ppm range for compound 3, and two doublet peaks at 7.54, 7.60 and 7.48, 7.60 ppm with coupling constants of 15.3 Hz for compounds 2 and 4, respectively. The 13C-NMR spectrum of prototypical compound 2 showed the two carbonyl carbons at 187.8 and 192.7 ppm.

2.1.2. Synthesis of Pyrazoline Derivatives 5–9 and Isoxazoline Derivatives 10–12

The compounds 24 were converted into the corresponding 3-(aryl)-5-(6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)-4,5-dihydro-1H-pyrazole-1-carbothioamides 57 by treatment with thiosemicarbazide (Scheme 1).

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Scheme 1. Synthesis of chalcones 24, pyrazoline derivatives 59 and isoxazoline derivatives 1012.

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Scheme 1. Synthesis of chalcones 24, pyrazoline derivatives 59 and isoxazoline derivatives 1012.
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Their 1H-NMR spectra showed multiplets within the 3.33–4.82 range corresponding to H4, H4' of the pyrazoline ring, where a multiplet at 6.92–6.98 ppm is observed for compound 7 corresponding to H5. A doublet of doublets at 6.73–6.88 corresponding to H5 of the pyrazoline ring was observed for compounds 5 and 6, respectively. In addition to a broad signal corresponding to the exchangeable NH2 protons was observed in the 7.25–8.01 ppm range. The 13C-NMR spectrum of compound 6 chosen as a prototype showed C=S and C=O peaks at 180.0 and 204.6 ppm, respectively. Reaction of compounds 2 and 3 with hydrazinum chloride gave rise to 2-(3-(aryl)-4,5-dihydro-1H-pyrazol-5-yl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-ones 8 and 9 (Scheme 1). In their 1H-NMR spectra, the appearance of signals in the ranges 3.33–3.75 and 5.49–6.70 ppm corresponding to (H4, H4') and H5 of the pyrazoline ring respectively was observed. The product of compound 4 with hydrazinium chloride could not be separated in a pure form. The 2-(3-(aryl)-4,5-dihydroisoxazol-5-yl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-ones 1012 were synthesized by cyclization of 24 in presence of hydroxylamine hydrochloride. Their 1H-NMR spectra showed three signals within the ranges 3.34–4.22 and 5.55–6.52 ppm corresponding to the (H4, H4') and H5 of the pyrazoline ring, respectively. The 13C-NMR spectrum of compound 11 selected as a prototype showed the carbonyl carbon at 193.7 ppm.

2.1.3. Synthesis of Benzenesulfonamide Derivatives 13–21

Reaction of the prepared chalcones 24 with 4-hydrazinyl benzenesulfonamide hydrochloride afforded 4-(3-(aryl)-5-(6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)-4,5-dihydro-1H-pyrazol-1-yl)benzenesulfonamides 1315 (Scheme 2).

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Scheme 2. Synthesis of benzenesulfonamide derivatives 1321.

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Scheme 2. Synthesis of benzenesulfonamide derivatives 1321.
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Their 1H-NMR spectra showed three signals at 3.15–3.40, 3.36–4.00, and 4.80–5.56 ppm corresponding to the H4, H4' , and H5 of the pyrazoline ring. In addition a broad singlet was observed in the 6.63–7.35 ppm range corresponding to the NH2 protons. The 13C-NMR spectrum of compound 15 as a prototype showed C=O at 193.8 ppm. On the other hand, the reaction of pyrazolines 1315 with bromine in acetic acid [68] at room temperature in order to obtain the pyrazoles, afforded the corresponding substituted 4-(4-bromo-3-(4-bromophenyl)-5-(6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)-1H-pyrazol-1-yl)benzenesulfonamide derivatives 1618, respectively in 79–84% yield.

Furthermore, the prepared compound 17 was treated with phenyl isothiocyanate to furnish 4-(4-bromo-3-aryl-5-(6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)-1H-pyrazol-1-yl)-N-(phenyl-carbamothioyl)benzene sulfonamide 19 in 76.8% yield (Scheme 2). The proton NMR spectrum showed three broad singlets at 8.62, 9.76, and 11.11 ppm corresponding to three NH protons.

The prepared substituted benzenesulfonamides 13 and 14 were allowed to react with phenyl isothiocyanate to correspondingly furnish 4-(3-(aryl)-5-(6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)-4,5-dihydro-1H-pyrazol-1-yl)-N-(phenylcarbamothioyl)benzenesulfonamides 20 and 21 (Scheme 2). Their 1H-NMR spectra showed D2O exchangeable signals at the ranges 9.00–9.15, 10.54–10.60, and 11.03–11.08 ppm, corresponding to three NH protons.

2.1.4. Synthesis of [1,2,4]Triazolo[3,4-c][1,2,4]triazino[5,6-b]-5-N-(phenylcarbamothioyl) Ethanoic Acid Hydrazide Derivatives 30, 31 and 3-Methyl-4-(propan-2-ylidene)-1H-pyrazol-5(4H)-one Derivatives 34, 35

Reaction of indoline-2,3-dione (1'') [69,70] with thiosemicarbazide gave rise to 5H-[1,2,4] triazino[5,6-b]indole-3-thiol (22) [70] (Scheme 3). The 1H-NMR spectrum showed two broad singlets exchangeable with D2O at 12.43 and 14.54 ppm, corresponding to the two NH protons, which confirm the structure of 22. The 13C-NMR also confirmed the structure of 22 with a peak at 179.5 corresponding to the C=S group. Treatment of the thiol derivative 22 with hydrazine hydrate afforded 3-hydrazinyl-5H-[1,2,4]triazino[5,6-b]indole (24) [70] in 92.6% yield (Scheme 3). The proton NMR spectrum showed two broad singlets at 4.31 and 8.54 ppm corresponding to NH2 and NH protons of hydrazine chain, in addition to a broad singlet at 11.82 ppm corresponding to the indole ring NH.

The reaction of 24 with diethyl malonate gave rise to the corresponding ester 26. The proton NMR spectrum showed a triplet signal at 1.16 ppm corresponding to the CH3 protons, and a quartet signal at 4.31 ppm corresponding to CH2 of the ester moiety, and a singlet signal at 4.12 corresponding to CH2 protons. Reaction of the ester 26 with hydrazine hydrate afforded the corresponding 5-ethanoic hydrazide 28 (Scheme 3). From the proton NMR spectrum the disappearance of CH3 and CH2 protons of the ester chain can be observed. Treatment of the prepared hydrazide 28 with phenyl isothiocyanate afforded the corresponding 5-N-(phenylcarbamothioyl)ethanoic acid hydrazide 30 in 80.1% yield (Scheme 3). Its structure was confirmed by 1H-NMR, 13C-NMR spectra, and elemental analysis. The 13C-NMR spectrum showed C=S and C=O carbons at 168.7 and 189.4 ppm respectively (see Experimental part).

On the other hand, treatment of the prepared hydrazine derivative 24 with ethyl acetoacetate in acetic acid afforded 1-(5H-[1,2,4]triazino[5,6-b]indol-3-yl)-3-methyl-1H-pyrazol-5(4H)-one (32) (Scheme 3). The proton NMR spectrum of this compound showed the CH3 protons as a singlet at 1.80 ppm, and the CH2 protons as a singlet at 2.43 ppm. 1-(5H-[1,2,4]Triazino[5,6-b]indol-3-yl)-3-methyl-4-(propan-2-ylidene)-1H-pyrazol-5(4H)-one (34) was prepared from the previous pyrazoline-5-one derivative 32 by its reaction with acetone (Scheme 3). The proton NMR spectrum showed two methyl protons as a singlet at 1.87 ppm.

Cyclization of quinoxaline-2,3(1H,4H)-dione (1''') with thiosemicarbazide afforded 5,10-dihydro-[1,2,4]triazino[5,6-b]quinoxaline-3-thiol (23) in good yield (Scheme 3). Its proton NMR spectrum showed two broad singlets, exchangeable with D2O, at 11.88 and 14.50 ppm, corresponding to the three NH protons, which confirm the structure of 23. Treatment of this thiol derivative 23 with hydrazine hydrate gave 3-hydrazinyl-5,10-dihydro-[1,2,4]triazino[5,6-b]quinoxaline (25, Scheme 3). Its proton NMR spectrum showed NH2 protons as a broad singlet signal at 4.55 ppm, in addition to three NH protons, see Experimental part. Treatment of the prepared hydrazine derivative 25 with diethyl malonate gave rise to the corresponding ester 27 (Scheme 3). The proton NMR spectrum showed the ester protons (CH3, CH2) as a triplet and a quartet signals at 1.09 and 4.09 ppm, respectively, in addition to CH2 protons at 4.66 ppm as a singlet signal.

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Scheme 3. Synthesis of [1,2,4]triazolo[3,4-c][1,2,4]triazino[5,6-b]-5-N-(phenylcarbamothioyl) ethanoic acid hydrazide derivatives 30, 31 and 3-methyl-4-(propan-2-ylidene)-1H-pyrazol-5(4H)-one derivatives 34, 35.

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Scheme 3. Synthesis of [1,2,4]triazolo[3,4-c][1,2,4]triazino[5,6-b]-5-N-(phenylcarbamothioyl) ethanoic acid hydrazide derivatives 30, 31 and 3-methyl-4-(propan-2-ylidene)-1H-pyrazol-5(4H)-one derivatives 34, 35.
Molecules 18 02683 g003 1024

Reaction of the ester 27 with hydrazine hydrate leads to the corresponding 5-ethanoic hydrazide 29. The proton NMR spectrum showed the disappearance of CH3 and CH2 protons of the ester chain, and NH and NH2 protons at 8.90 and 9.61 ppm were observed as a two broad singlets. Its 13C-NMR spectrum showed the carbonyl carbon at 168.4 ppm. Treatment of the prepared hydrazide 29 with phenyl isothiocyanate leads to corresponding 5-N-(phenylcarbamothioyl) ethanoic acid hydrazide 31 (Scheme 3). Its structure was also confirmed by 1H-NMR, and elemental analysis.

Reaction of the hydrazine derivative 25 with ethyl acetoacetate in acetic acid afforded 1-(5,10-dihyro-[1,2,4]triazino[5,6-b]quinoxalin-3-yl)-3-methyl-1H-pyrazol-5(4H)-one (33, Scheme 3). The proton NMR spectrum showed the CH3 protons at position 3 of the pyrazoline ring as a singlet signal at 2.29 ppm, and the CH2 protons (H4) of the pyrazoline ring as a singlet signal at 2.92 ppm, with the disappearance of the peak corresponding to NH2 protons. 13C-NMR spectrum showed the carbonyl carbon at 170.0 ppm. Reaction of the prepared pyrazoline-5-one 33 with acetone gave rise to 1-(5,10-dihydro-[1,2,4]triazino[5,6-b]quinoxalin-3-yl)-3-methyl-4-(propan-2-ylidene)-1H-pyrazol-5(4H)-one (35, Scheme 3). Its proton NMR spectrum showed three methyl groups at 2.30, 2.42, and 2.93 ppm.

2.1.5. Synthesis of Schiff’s Bases 36, 37, 4-Oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)thiazolidin-4-one (38), Benzohydrazide Derivative 39, 1,2,4]Triazolo[3,4-c]-5,10-dihydro [1,2,4]triazino[5,6-b] quinoxaline (40), and Pyrazole Derivatives 41–44

Condensation of hydrazine derivative 24 with 6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indole-2-carbaldehyde (1') afforded the corresponding Schiff's base 36 (Scheme 4). Its proton NMR spectrum showed the disappearance of the NH2 signal, and a singlet signal corresponding to a CH=N proton at 8.03 ppm was observed. The 13C-NMR spectrum showed the carbonyl carbon at 192.6 ppm. Treatment of the prepared compound 36 with thioglycolic acid in dry benzene gave rise to corresponding 3-(5H-[1,2,4]triazino[5,6-b]indol-3-yl)-2-(6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl) thiazolidin-4-one (38, Scheme 4). The proton NMR spectrum showed the CH2 (H5, H5') protons of the thiazolidine ring at 3.35–3.49 ppm as a multiplet and the CH proton of thiazolidine ring (H2) as a singlet signal at 7.93 ppm.

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Scheme 4. Synthesis of Schiff’s bases 36, 37, 4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)thiazolidin-4-one (38), benzohydrazide derivative 39, 1,2,4]triazolo[3,4-c]-5,10-dihydro [1,2,4]triazino[5,6-b] quinoxaline (40), and pyrazole derivatives 4144.

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Scheme 4. Synthesis of Schiff’s bases 36, 37, 4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)thiazolidin-4-one (38), benzohydrazide derivative 39, 1,2,4]triazolo[3,4-c]-5,10-dihydro [1,2,4]triazino[5,6-b] quinoxaline (40), and pyrazole derivatives 4144.
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Condensation of the hydrazine derivative 25 with 6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indole-2-carbaldehyde (1') furnished to the corresponding Schiff's base 37 in 86.6% yield (Scheme 4). Its proton NMR spectrum showed the CH=N proton as a singlet at 9.45 ppm. Condensation of 37 with benzoyl hydrazine afforded the corresponding N'-2-((2-(5,10-dihydro-[1,2,4]triazino[5,6-b]quinoxalin-3-yl)hydrazono)methyl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-ylidene)benzohydrazide (39). Its 13C-NMR spectrum showed the C=O group at 162.7 ppm. On the other hand, oxidative cyclization of Schiff's base 37 with acetic anhydride afforded the cyclized compound 40 (Scheme 4). The proton NMR spectrum showed the disappearance of CH=N proton. We expected to obtain the acetylated product, but the 1H-NMR spectrum confirmed the structure of the cyclized compound 40 as shown, with an exchangeable peak corresponding to three NH protons being observed at 12.23 ppm, and no peak observed corresponding to the acetyl methyl group. Oxidative cyclization of 3-hydrazinyl-5H-[1,2,4]triazino[5,6-b]indole (24) with 2-(3-(4-bromophenyl)-3-oxoprop-1-enyl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one (2) afforded 2-(1-(5H-[1,2,4]triazino[5,6-b]indol-3-yl)-3-(4-bromo-phenyl)-1H-pyrazol-5-yl)-(6,6-dimethyl-6,7-dihydro-1H-indol-4 (5H)-one (41) in 80.8% yield. The proton NMR spectrum showed the CH proton of pyrazole ring as a singlet signal at 6.12 ppm. The 13C-NMR spectrum showed C=O carbon 190.8 at ppm.

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Scheme 5. Charge distribution on nitrogen atoms N1, N3 of compound 37.

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Scheme 5. Charge distribution on nitrogen atoms N1, N3 of compound 37.
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On the other hand, the hydrazine derivative 25 was allowed to react with the prepared chalcones 24 to correspondingly furnish 2-(3-(aryl)-1-(5,10-dihydro-[1,2,4]triazino[5,6-b]quinoxalin-3-yl)-1H-pyrazol-5-yl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-ones 4244, respectively (Scheme 4). Their proton NMR spectra showed the CH protons of the pyrazole ring (H4) and indole ring (H3) at 6.80 and 6.91, 6.90 and 7.27 and 6.78 and 6.85, ppm respectively. The 13C-NMR spectra of compounds 4244 showed the C=O of the indole ring at 192.8–193.3 ppm. According to the charge distribution determined using ChemDraw Ultra, the N1 nitrogen atom has a better nucleophile character compared to the N3 nitrogen atom, which is in accordance with the proposed structure of compound 40 (Scheme 5).

2.2. Pharmacological Screening

Four test organisms representing different groups of microorganisms were used to evaluate the bioactivity of the designed products. The utilized test organisms were: Escherichia coli ATCC8739, Pseudomonas aeruginosa ATCC 9027 as Gram-negative bacteria, Staphylococcus aureus ATCC 6538P as an example of Gram-positive bacteria, and Candida albicans ATCC 2091 as yeast-like fungi. The inhibition zone (IZ) and minimal inhibitory zone (MIC) results are given in Table 1.

Table 1. In vitro antimicrobial activity of the test compounds and evaluation of the inhibition zone (IZ) and the minim. inhibitory concentration (MIC).

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Table 1. In vitro antimicrobial activity of the test compounds and evaluation of the inhibition zone (IZ) and the minim. inhibitory concentration (MIC).
MicroorganismEscherichia coliStaphylococcus aureusCandida albicansPseudomonas aeruginosa
IZMICIZMICIZMICIZMIC
ampicillin 10.0 µg/disc18252212.5---------------------
ciprofloxacin 5.0 µg/disc2812.53025-----------3825
clotrimazole 100.0 µg/disc--------------------4012.5-----------
imipenam 10.0 µg/disc26-----30---------------30------
218200172002120016200
319200172002420018200
419200152002320018200
519200152002320019200
619200162002520020200
718200132002120016200
819200172002220018200
918200162002420018200
1019200162002420020200
1119200181002320019200
1219200152002320018200
1318200192002320018200
1418200152002320018200
1518200161002220019200
16192002625275020200
1719200>50100>405021100
1819200201002320019200
1919200161002512.519200
2018200162002612.518200
211820016200285020200
221920016200255018200
231820082002420020200
2418200162002320020200
2519200152002220018200
2619200132002320019200
2719200172002520017200
2819200172002320018200
2918200152002120018200
3019200172002620016200
311920016200342520200
3219200162002320020200
3319200172002420018200
3419200172002420017200
3519200161002320019200
3619200172002420018200
3717200152002220018200
3819200172002120018200
3919200172002220016200
4019200172002120018200
4118200132002120016200
4218200162002320018200
4318200171002420019200
4420200171002320019200
DMF18 13 21 16

The compounds under investigation 244 did not show any activity against the test organisms Escherichia coli and Pseudomonas aeruginosa. The inhibition Zone (IZ) listed in Table 1 showed that compound 16 has good antimicrobial activity against Staphylococcus aureus, comparable to that of ampicillin, while compound 17 has remarkable antimicrobial activity against Staphylococcus aureus exceeding that of ampicillin, ciprofloxacin and imipenam.. The minimal inhibitory concentration (MIC) value showed that compound 16 has good antimicrobial activity against Staphylococcus aureus, comparable to that of ciprofloxacin, while its activity is about 50% of that of ampicillin. In addition, compound 17 has an IZ against Candida albicans comparable to that of clotrimazole. The minimal inhibitory concentration (MIC) of compound 17 against Candida albicans is about 25% of that clotrimazole. On the other hand, the minimal inhibitory concentration (MIC) of compounds 19 and 20 against Candida albicans was good, and comparable to that of clotrimazole, while compound 31 has 50 % activity compared to that of clotrimazole.

3. Experimental

3.1. General Methods

Fresh solvents were used without purification. Melting points were obtained in open capillary tubes by using a MEL-Temp II melting point apparatus and are uncorrected. Infrared spectra (IR) were recorded on a Perkin-Elmer 1600 series Fourier Transform instrument with the samples as KBr pellets. 1H-NMR and 13C-NMR spectra were recorded on a JEOL 500 MHz spectrometer at ambient temperature using tetramethylsilane as an internal reference. Elemental analyses were carried out by the University of Cairo Microanalytical Laboratories. The antimicrobial tests were carried out at the Pharmaceutical Chemistry Department, Faculty of Pharmacy, Alexandria University. ChemDraw-Ultra-11.0 has been used for the nomenclature of the prepared compounds.

3.2. General Procedure for the Preparation of Compounds 2–4

An equimolar mixture of 6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indole-2-carbaldehyde (1', 1.91 g, 0.01 mol) [67] and the substituted acetophenone (0.01 mol) in 2% ethanolic KOH (20 mL) was stirred at room temperature (R.T.) for 5 h. The solid product was cooled, collected by filtration, washed with water, dried and recrystallized from chloroform/ethanol.

2-(3-(4-Bromophenyl)-3-oxoprop-1-enyl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one (2). Yellow crystals; yield 3.36 g, 90.5%; m.p. 269–270 °C; IR (KBr): 1651 (C=O), 3252 cm−1 (NH); 1H-NMR (DMSO-d6) δ 1.01 (s, 6H, 2 CH3), 2.23 (s, 2H, CH2), 2.72 (s, 2H, CH2), 6.91 (s, 1H, CH-pyrrole), 7.54 (d, 1H, COCH=; J = 15.3 Hz), 7.60 (d, 1H, CH=; J = 15.3 Hz), 7.76 (d, 2H, ArH; J = 8.4 Hz), 7.93 (d, 2H, ArH; J = 8.4 Hz), 12.14 (bs, 1H, NH; exchangeable with D2O); 13C-NMR (DMSO-d6) δ:28.6, 35.7, 36.5, 52.2, 113.5, 116.7, 121.0, 127.5, 130.6, 132.4, 134.6, 137.5, 147.8, 187.8, 192.7. Anal. Calcd for C19H18BrNO2 (372.26): C, 61.30; H, 4.87; N, 3.76 Found: C, 61.42; H, 4.97; N, 3.53.

2-(3-(4-Chlorophenyl)-3-oxoprop-1-enyl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one (3). Yellow crystals; yield 2.99 g, 91.5%; m.p. 261–262 °C; IR (KBr): 1651 (C=O), 3250 cm−1 (NH); 1H-NMR (DMSO-d6) δ 1.01 (s, 6H, 2 CH3), 2.23 (s, 2H, CH2), 2.72 (s, 2H, CH2), 6.91 (s, 1H, CH-pyrrole), 7.52–7.63 (m, 4H, CH=CH, 2 ArH), 8.01 (d, 2H, ArH; J = 8.4 Hz), 12.12 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C19H18ClNO2 (327.80): C, 69.62; H, 5.53; N, 4.27 Found: C, 69.82; H, 5.76; N, 3.99.

2-(3-(4-Methoxyphenyl)-3-oxoprop-1-enyl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one (4). Yellow crystals; yield 3.00 g, 93.0%; m.p. 240–241 °C; IR (KBr): 1651 (C=O), 3232 cm−1 (NH); 1H-NMR (DMSO-d6) δ 1.01 (s, 6H, 2 CH3), 2.23 (s, 2H, CH2), 2.71 (s, 2H, CH2), 3.82 (s, 3H, OCH3), 6.85 (s, 1H, CH-pyrrole), 7.07 (d, 2H, ArH; J = 8.4 Hz), 7.48 (d, 1H, -COCH=; J = 15.3 Hz), 7.60 (d, 1H, CH=; J = 15.3 Hz), 8.01 (d, 2H, ArH; J = 8.4 Hz), 12.03 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C20H21NO3 (323.39): C, 74.28; H, 6.55; N, 4.33 Found: C, 74.40; H, 6.71; N, 4.11.

3.3. General Procedure for the Preparation of Compounds 5–7

A mixture of the appropriate 2-(3-aryl-3-oxoprop-1-enyl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one 24 (0.002 mol) and thiosemicarbazide (0.003 mol) was dissolved in a mixture of acetone and ethanol (30 mL), then potassium carbonate (0.004 mol) was added with vigorous stirring. Heating under reflux was performed for 14 h. The solvent was removed under vacuum, and ice-water was added to the reaction mixture. The solid product obtained was filtered, washed with ethanol, dried and recrystallized from chloroform/ethanol.

3-(4-Bromophenyl)-5-(6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)-4,5-dihydro-1H-pyrazole-1-carbothioamide 5. Buff crystals; yield 0.71 g, 80.0%; m.p. 192–193 °C; IR (KBr): 1167 (C=S), 1651 (C=O), 3134, 3232, 3436 cm−1 (NH, NH2); 1H-NMR (DMSO-d6) δ 1.13 (s, 6H, 2 CH3), 2.39 (s, 2H, CH2), 2.80 (s, 2H, CH2), 3.86–3.89 (m, 2H, H4, H4'-pyrazoline), 6.73–6.88 (dd, 1H, H5-pyrazoline; J = 17.4, 12.0 Hz), 6.92 (s, 1H, CH-pyrrole), 6.97 (d, 2H, ArH; J = 8.6 Hz), 7.25 (bs, 2H, NH2; exchangeable with D2O), 8.01 (d, 2H, ArH; J = 8.6 Hz), 12.05 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C20H21BrN4OS (445.38): C, 53.94; H, 4.75; N, 12.58 Found: C, 53.70; H, 4.60; N, 12.76.

3-(4-Chlorophenyl)-5-(6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)-4,5-dihydro-1H-pyrazole-1-carbothioamide 6. Buff crystals; yield 0.63 g, 79.8%; m.p. 210–211 °C; IR (KBr): 1092 (C=S), 1646 (C=O), 3237, 3428 cm−1 (NH, NH2); 1H-NMR (DMSO-d6) δ 0.95 (s, 3H, CH3), 1.02 (s, 3H, CH3), 2.24 (s, 2H, CH2), 2.72 (s, 2H, CH2), 3.33–3.41 (m, 2H, H4, H4’-pyrazoline), 6.80–6.88 (dd, 1H, H5-pyrazoline; J = 17.4, 12.0 Hz), 7.58 (s, 1H, CH-pyrrole), 7.47 (bs, 2H, NH2; exchangeable with D2O), 7.77 (d, 2H, ArH; J = 7.5 Hz), 7.93 (d, 2H, ArH; J = 7.5 Hz), 12.00 (bs, 1H, NH; exchangeable with D2O); 13C-NMR (DMSO-d6) δ: 29.0, 31.4, 40.4, 41.0, 52.5, 63.5, 114.4, 120.2, 125.3, 129.3, 129.5, 130.6, 136.0, 140.0, 150.49, 180.0, 204.6. Anal. Calcd for C20H21ClN4OS (400.92): C, 59.91; H, 5.28; N, 13.97 Found: C, 59.70; H, 5.11; N, 14.12.

5-(6,6-Dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)-3-(4-methoxyphenyl)-4,5-dihydro-1H-pyrazole-1-carbothioamide 7. Yellow crystals; yield 0.60 g, 76.7%; m.p. 259–260 °C; IR (KBr): 1167 (C=S), 1651 (C=O), 3134, 3232, 3436 cm−1 (NH, NH2); 1H-NMR (DMSO-d6) δ 1.13 (s, 6H, 2 CH3), 2.39 (s, 2H, CH2), 2.80 (s, 2H, CH2), 3.86–3.89 (m, 4H, H4-pyrazoline, OCH3), 4.73–4.82 (m, 1H, H4'-pyrazoline), 6.92–6.98 (m, 1H, H5-pyrazoline), 7.15–7.25 (m, 5H, 4 ArH, CH-pyrrole), 8.01 (bs, 2H, NH2; exchangeable with D2O), 11.59 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C21H24N4O2S (396.51): C, 63.61; H, 6.10; N, 14.13 Found: C, 63.80; H, 6.24; N, 13.90.

3.4. General Procedure for the Preparation of Compounds 8 and 9

A mixture of 2-(3-aryl-3-oxoprop-1-enyl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one 2 or 3 (0.001 mol), hydrazinum chloride (0.003 mol) and anhydrous sodium acetate (0.003 mol), in ethanol (15 mL) and glacial acetic acid (5 mL) was refluxed for 8 h. The reaction mixture was poured over crushed ice. The solid obtained was filtered, washed with water, dried and recrystallized from ethanol.

2-(3-(4-Bromophenyl)-4,5-dihydro-1H-pyrazol-5-yl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one (8). Brown crystals; yield 0.31 g, 81.3%; m.p. 210–211 °C; IR (KBr): 1641 (C=O), 3260, 3436 cm−1 (NH); 1H-NMR (DMSO-d6) δ 0.98 (s, 6H, 2 CH3), 2.24 (s, 2H, CH2), 2.72 (s, 2H, CH2), 3.33–3.75 (m, 2H, H4, H4'-pyrazoline), 6.50–6.70 (m, 1H, H5-pyrazoline), 6.91 (s, 1H, CH-pyrrole), 7.77 (d, 2H, ArH; J = 7.7 Hz), 7.93 (d, 2H, ArH; J = 7.7 Hz), 11.25 (bs, 1H, NH; exchangeable with D2O), 12.08 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C19H20BrN3O (386.29): C, 59.08; H, 5.22; N, 10.88 Found: C, 59.26; H, 5.45; N, 10.65.

2-(3-(4-Chlorophenyl)-4,5-dihydro-1H-pyrazol-5-yl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one (9). Buff crystals; yield 0.26 g, 78.8%; m.p. 195–196 °C; IR (KBr): 1642 (C=O), 3209, 3423 cm−1 (NH); 1H-NMR (DMSO-d6) δ 0.98 (s, 3H, CH3), 1.01 (s, 3H, CH3), 2.20 (s, 2H, CH2), 2.42 (s, 2H, CH2), 3.53–3.75 (m, 1H, H4-pyrazoline), 3.69–3.75 (m, 1H, H4'-pyrazoline), 5.50 (m, 1H, H5-pyrazoline), 6.69 (s, 1H, CH-pyrrole), 7.41 (d, 2H, ArH; J = 7.7 Hz), 7.78 (d, 2H, ArH; J = 7.7 Hz), 11.19 (bs, 1H, NH; exchangeable with D2O), 12.07 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C19H20ClN3O (341.83): C, 66.76; H, 5.90; N, 12.29 Found: C, 66.53; H, 5.69; N, 12.50.

3.5. General Procedure for the Preparation of Compounds 10–12

A mixture of 2-(3-aryl-3-oxoprop-1-enyl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-ones 24 (0.001 mol), hydroxylamine hydrochloride (0.003 mol) and anhydrous sodium acetate (0.003 mol), in ethanol (15 mL) and glacial acetic acid (5 mL) was refluxed for 8 h. The reaction mixture was poured over crushed ice. The solid obtained was filtered, washed with water, dried and recrystallized from ethanol.

2-(3-(4-Bromophenyl)-4,5-dihydroisoxazol-5-yl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one (10). Yellow crystals; yield 0.34 g, 90.1%; m.p. 115–116 °C; IR (KBr): 1646 (C=O), 3423 cm−1 (NH); 1H-NMR (DMSO-d6) δ 0.95 (s, 6H, 2 CH3), 2.42 (s, 2H, CH2), 2.51 (s, 2H, CH2), 3.34–3.46 (m, 2H, H4, H4'-isoxazoline), 6.50–6.52 (m, 1H, H5 isoxazoline), 6.84 (s, 1H, CH-pyrrole), 7.49–7.56 (m, 4H, ArH), 11.32 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C19H19BrN2O2 (387.27): C, 58.93; H, 4.95; N, 7.23 Found: C, 58.75; H, 4.86; N, 7.39.

2-(3-(4-Chlorophenyl)-4,5-dihydroisoxazol-5-yl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one (11). Buff crystals; yield 0.26 g, 78.3%; m.p. 160–161 °C; IR (KBr): 1643 (C=O), 3433 cm−1 (NH); 1H-NMR (DMSO-d6) δ 1.01 (s, 6H, 2 CH3), 2.35 (s, 2H, CH2), 2.59 (s, 2H, CH2), 3.50–3.55 (m, 1H, H4-isoxazoline), 3.66–3.72 (m, 1H, H4'-isoxazoline), 5.66–5.70 (m, 1H, H5 isoxazoline), 6.81 (s, 1H, CH-pyrrole), 7.51 (d, 2H, ArH; J = 6.7 Hz), 7.70 (d, 2H, ArH; J = 6.7 Hz), 11.97 (bs, 1H, NH; exchangeable with D2O). 13C-NMR (DMSO-d6) δ: 27.9, 34.4, 36.6, 43.3, 55.0, 76.8, 108.1, 109.8, 128.1, 128.9, 129.4, 131.9, 135.2, 143.6, 156.5, 193.7. Anal. Calcd for C19H19ClN2O2 (342.82): C, 66.57; H, 5.59; N, 8.17 Found: C, 66.40; H, 5.35; N, 8.34.

2-(3-(4-Methoxyphenyl)-4,5-dihydroisoxazol-5-yl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one (12). Buff crystals; yield 0.27 g, 81.0%; m.p. 170–171 °C; IR (KBr): 1645 (C=O), 3427 cm−1 (NH); 1H-NMR (DMSO-d6) δ 0.98 (s, 6H, 2 CH3), 2.46 (s, 4H, 2 CH2), 3.34 (s, 3H, OCH3), 3.46–3.63 (m, 1H, H4-isoxazoline), 4.08–4.22 (m, 1H, H4'-isoxazoline), 5.55–5.66 (m, 1H, H5 isoxazoline), 6.72 (s, 1H, CH-pyrrole), 6.98 (d, 2H, ArH; J = 6.9 Hz), 7.62 (d, 2H, ArH; J = 6.9 Hz), 11.16 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C20H22N2O3 (338.40): C, 70.99; H, 6.55; N, 8.28 Found: C, 70.79; H, 6.31; N, 8.47.

3.6. General Procedure for the Preparation of Compounds 13–15

A mixture of 2-(3-aryl-3-oxoprop-1-enyl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one 24 (0.001 mol) and 4-hydrazinylbenzenesulfonamide hydrochloride (0.001 mol) in methanol (30 mL) was heated under reflux for 6 h, partially concentrated and cooled. The separated solid product was filtered, washed with ethanol, dried and recrystallized from ethanol.

4-(3-(4-Bromophenyl)-5-(6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)-4,5-dihydro-1H-pyrazol-1-yl)benzenesulfonamide (13). Brown crystals; yield 0.48 g, 90.2%; mp 180–181 °C; IR (KBr): 1219, 1374 (SO2), 1651 (C=O), 3250, 3434 cm−1 (NH, NH2); 1H-NMR (DMSO-d6) δ 0.93 (s, 3H, CH3), 1.01 (s, 3H, CH3), 2.50 (s, 2H, CH2), 2.66 (s, 2H, CH2), 3.15–3.22 (m, 1H, H4-pyrazoline), 3.36–3.40 (m, 1H, H4'-pyrazoline), 5.53–5.56 (m, 1H, H5- pyrazoline), 6.43 (s, 1H, CH-pyrrole), 6.63 (bs, 2H, NH2; exchangeable with D2O), 7.00–7.26 (m, 2H, ArH), 7.40–7.70 (m, 2H, ArH), 7.73–7.93 (m, 2H, ArH), 8.05–8.18 (m, 2H, ArH), 11.61 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C25H25BrN4O3S (541.46): C, 55.46; H, 4.65; N, 10.35 Found: C, 55.60; H, 4.87; N, 10.15.

4-(3-(4-Chlorophenyl)-5-(6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)-4,5-dihydro-1H-pyrazol-1-yl)benzenesulfonamide (14). Brown crystals; yield 0.43 g, 87.5%; m.p. 150–151 °C; IR (KBr): 1219, 1374 (SO2), 1651 (C=O), 3256, 3438 cm−1 (NH, NH2); 1H-NMR (DMSO-d6) δ 1.01 (s, 6H, 2 CH3), 2.24 (s, 2H, CH2), 2.72 (s, 2H, CH2), 3.20–3.40 (m, 1H, H4-pyrazoline), 3.78–3.84 (dd, 1H, H4'-pyrazoline; J = 17.4, 12.0 Hz), 5.53–5.55 (m, 1H, H5-pyrazoline), 6.85 (s, 1H, CH-pyrrole), 7.13 (d, 2H, ArH; J = 9.1 Hz), 7.34 (bs, 2H, NH2; exchangeable with D2O), 7.43 (d, 2H, ArH; J = 9.1 Hz), 7.53 (d, 2H, ArH; J = 8.4 Hz), 7.69 (d, 2H, ArH; J = 8.4 Hz), 12.18 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C25H25ClN4O3S (497.01): C, 60.41; H, 5.07; N, 11.27 Found: C, 60.66; H, 5.30; N, 11.03.

4-(5-(6,6-Dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)-3-(4-methoxyphenyl)-4,5-dihydro-1H-pyrazol-1-yl)benzenesulfonamide (15). Buff crystals; yield 0.44 g, 91.1%; m.p. 224–225 °C; IR (KBr): 1158, 1307 (SO2), 1650 (C=O), 3263, 3345 cm−1 (NH, NH2); 1H-NMR (DMSO-d6) δ 1.01 (s, 6H, 2 CH3), 2.20 (s, 2H, CH2), 2.68 (s, 2H, CH2), 3.24–3.28 (m, 1H, H4-pyrazoline), 3.34 (s, 3H, OCH3), 3.76–4.00 (m, 1H, H4'-pyrazoline), 4.80–5.53 (m, 1H, H5-pyrazoline), 6.46 (s, 1H, CH-pyrrole), 7.00–7.04 (m, 4H, 2 ArH, NH2), 7.13 (d, 2H, ArH; J = 8.4 Hz), 7.60 (d, 2H, ArH; J = 8.4 Hz), 7.70–7.73 (m, 2H, ArH), 11.65 (bs, 1H, NH; exchangeable with D2O). 13C-NMR (DMSO-d6) δ: 28.7, 35.1, 40.9, 43.7, 53.6, 55.7, 60.3, 106.4, 114.7, 118.4, 119.9, 125.4, 127.6, 127.9, 130.3, 141.6, 142.3, 153.2, 160.0, 193.8. Anal. Calcd for C26H28N4O4S (492.59): C, 63.40; H, 5.73; N, 11.37 Found: C, 63.69; H, 5.90; N, 11.22.

3.7. General Procedure for the Preparation of Compounds 16–18

Solid 4-(3-aryl-5-(6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)-4,5-dihydro-1H-pyrazol-1-yl)benzenesulfonamide 1315 (0.001 mol) was dissolved in hot acetic acid, and after cooling bromine (0.3 mL) in acetic acid (10 mL) was added dropwise with shaking, then the reaction mixture was allowed to stand at R.T. overnight. The solid mass obtained was separated by filtration, washed with water, dried and recrystallized from ethanol.

4-(4-Bromo-3-(4-bromophenyl)-5-(6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)-1H-pyrazol-1-yl)benzenesulfonamide (16). Buff crystals; yield 0.49 g, 79.3%; m.p. 200–201 °C; IR (KBr): 1162, 1335 (SO2), 1653 (C=O), 3253, 3433 cm−1 (NH, NH2); 1H-NMR (DMSO-d6) δ 1.09 (s, 6H, 2 CH3), 2.26 (s, 2H, CH2), 2.86 (s, 2H, CH2), 3.58–3.88 (m, 1H, H4-pyrazoline), 4.80–5.00 (m, 1H, H5 -pyrazoline), 7.44–7.46 (m, 3H, CH-pyrrole, NH2), 7.71 (d, 2H, ArH; J = 7.7 Hz), 7.86 (d, 2H, ArH; J = 8.4 Hz), 7.92 (d, 2H, ArH; J = 8.4 Hz), 8.07 (d, 2H, ArH; J = 7.7 Hz), 9.02 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C25H24Br2N4O3S (620.36): C, 48.40; H, 3.90; N, 9.03 Found: C, 48.80; H, 3.76; N, 8.87.

4-(4-Bromo-3-(4-chlorophenyl)-5-(6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)-1H-pyrazol-1-yl)benzenesulfonamide (17). Brown crystals; yield 0.47 g, 82.4%; m.p. 129–130 °C; IR (KBr): 1163, 1315 (SO2), 1670 (C=O), 3068, 3385 cm−1 (NH, NH2); 1H-NMR (DMSO-d6) δ 1.16 (s, 6H, 2 CH3), 2.27 (s, 2H, CH2), 2.47 (s, 2H, CH2), 3.13–3.73 (m, 1H, H4-pyrazoline), 3.88–4.16 (m, 1H, H5-pyrazoline), 7.56 (s, 1H, H-pyrrole), 7.63 (bs, 2H, NH2; exchangeable with D2O), 7.80–7.85 (m, 4H, ArH), 8.00–8.10 (m, 4H, ArH), 11.06 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C25H24BrClN4O3S (575.91): C, 52.14; H, 4.20; N, 9.73 Found: C, 52.25; H, 4.00; N, 9.58.

4-(4-Bromo-5-(6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)-3-(4-methoxy-phenyl)-1H-pyrazol-1-yl)benzenesulfonamide (18). Brown crystals; yield 0.48 g, 84.3%; m.p. 160–161 °C; IR (KBr): 1164, 1340 (SO2), 1650 (C=O), 3220, 3435 cm−1 (NH, NH2); 1H-NMR (DMSO-d6) δ 1.01 (s, 3H, CH3), 1.02 (s, 3H, CH3), 2.26 (s, 2H, CH2), 2.70 (s, 2H, CH2), 3.34 (s, 3H, OCH3), 3.80–3.89 (m, 2H, H4, H5-pyrazoline), 6.93 (d, 1H, ArH; J = 8.4 Hz), 7.03–7.07 (m, 2H, ArH, CH-pyrrole), 7.42 (d, 2H, ArH; J = 9.1 Hz), 7.82 (d, 2H, ArH; J = 9.1 Hz), 7.87–7.93 (m, 4H, 2ArH, NH2), 12.25 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C26H27BrN4O4S (571.49): C, 54.64; H, 4.76; N, 9.80 Found: C, 55.01; H, 4.65; N, 9.68.

3.8. 4-(4-Bromo-3-aryl-5-(6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)-1H-pyrazol-1-yl)-N-(phenylcarbamothioyl)benzenesulfonamide (19)

A mixture of 4-(4-bromo-3-(4-chlorophenyl)-5-(6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)-1H-pyrazol-1-yl) benzenesulfonamide (17, 0.005 mol) and anhydrous potassium carbonate (0.01 mol) in dry acetone (100 mL) was stirred under reflux for 15 h. A solution of phenyl isothiocyanate (0.007 mol) in dry acetone was added drop by drop at this temperature, and refluxing was continued for 12 h more. The acetone was distilled under reduced pressure and the solid residue was dissolved in water, the product was isolated after acidification with 2 N HCl. The solid mass obtained was separated by filtration, washed with water, dried and recrystallized from ethanol. The product was obtained as buff crystals; yield 2.73 g, 76.8%; m.p. 92–93 °C; IR (KBr): 1092 (C=S), 1160, 1345 (SO2), 1648 (C=O), 3435 cm−1 (NH); 1H-NMR (DMSO-d6) δ 1.19 (s, 6H, 2 CH3), 1.99 (s, 2H, CH2), 2.61 (s, 2H, CH2), 3.87–4.03 (m, 1H, H4-pyrazoline), 6.90–6.92 (m, 1H, H5-pyrazoline), 7.09 (s, 1H, CH-pyrrole), 7.22–7.27 (m, 2H, ArH), 7.29–7.32 (m, 3H, ArH), 7.40–7.45 (m, 4H, ArH), 7.85 (d, 2H, ArH; J = 8.4 Hz), 8.06 (d, 2H, ArH; J = 8.4 Hz), 8.62 (bs, 1H, NH; exchangeable with D2O), 9.76 (bs, 1H, NH; exchangeable with D2O), 11.11 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C32H29BrClN5O3S2(711.09): C, 54.05; H, 4.11; N, 9.85 Found: C, 54.00; H, 3.96; N, 10.04.

3.9. General Procedure for the Preparation of Compounds 20 and 21

A mixture of 4-(3-aryl-5-(6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)-4,5-dihydro-1H-pyrazol-1-yl)benzenesulfonamide 13 and 14 (0.005 mol) and anhydrous potassium carbonate (0.01 mol) in dry acetone (100 mL) was stirred with refluxing for 15 h. A solution of phenyl isothiocyanate (0.007 mol) in dry acetone was added drop by drop at this temperature, and refluxing was continued for 12 h. The acetone was distilled under reduced pressure and the solid residue was dissolved in water, the product was isolated after acidification with 2 N HCl. The solid mass obtained was separated by filtration, washed with water, dried and crystallized from ethanol.

4-(3-(4-Bromophenyl)-5-(6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)-4,5-dihydro-1H-pyrazol-1-yl)-N-(phenylcarbamothioyl)benzenesulfonamide (20). the product was obtained as brown crystals; Yield 2.86 g, 84.6%; m.p. 210–211 °C; IR (KBr): 1089 (C=S), 1160, 1355 (SO2), 1646 (C=O), 3436 cm−1 (NH); 1H-NMR (DMSO-d6) δ 1.19 (s, 6H, 2 CH3), 2.69 (s, 2H, CH2), 2.85 (s, 2H, CH2), 3.50–3.60 (m, 1H, H4-pyrazoline), 4.08–4.15 (m, 1H, H4'-pyrazoline), 4.40–4.55 (m, 1H, H5-pyrazoline), 6.57 (s, 1H, CH-pyrrole), 7.10–7.18 (m, 2H, ArH), 7.20–7.33 (m, 2H, ArH), 7.40–7.55 (m, 1H, ArH), 7.60–7.72 (m, 2H, ArH), 7.80–7.93(m, 2H, ArH), 7.95–8.18 (m, 2H, ArH), 8.61–8.68 (m, 2H, ArH), 9.15 (bs, 1H, NH; exchangeable with D2O), 10.54 (bs, 1H, NH; exchangeable with D2O), 11.03 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C32H30BrN5O3S2 (676.65): C, 56.80; H, 4.47; N, 10.35 Found: C, 56.62; H, 4.26; N, 10.54.

4-(3-(4-Chlorophenyl)-5-(6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)-4,5-dihydro-1H-pyrazol-1-yl)-N-(phenylcarbamothioyl)benzenesulfonamide (21). Brown crystals; yield 2.49 g, 78.9%; m.p. 168–169 °C; IR (KBr): 1086 (C=S), 1160, 1355 (SO2), 1653 (C=O), 3434 cm−1 (NH); 1H-NMR (DMSO-d6) δ 1.16 (s, 6H, 2 CH3), 2.71 (s, 2H, CH2), 2.87 (s, 2H, CH2), 3.89–4.07 (m, 1H, H4-pyrazoline), 4.40–4.50 (m, 1H, H4'-pyrazoline), 6.90–6.94 (m, 1H, H5-pyrazoline), 7.02 (s, 1H, CH-pyrrole), 7.11–7.23 (m, 3H, ArH), 7.40–7.43 (m, 4H, ArH), 7.90–7.94 (m, 4H, ArH), 8.66–8.73 (m, 2H, ArH), 12.41 (bs, 2H, 2NH; exchangeable with D2O), 12.73 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C32H30ClN5O3S2 (632.20): C, 60.79; H, 4.78; N, 11.08 Found: C, 60.56; H, 4.62; N, 10.87.

3.10. 5H-[1,2,4]Triazino[5,6-b]indole-3-thiol (22)

A mixture of indoline-2,3-dione [69,70] (1'', 0.1 mol), thiosemicarbazide (0.11 mol) and anhydrous potassium carbonate (0.15 mol) [71] were stirred in water (500 mL) for 2 h at R.T., and then refluxed for 5 h. The mixture was cooled, filtered, and the filtrate was acidified with acetic acid. The solid mass obtained was separated by filtration, washed with water and dried. The product was recrystallized from ethanol. The product was obtained as yellow crystals; yield 19.29 g, 95.4%; m.p. 334–335 °C; IR (KBr): 3423 cm−1 (NH); 1H-NMR (DMSO-d6) δ 7.27 (t, 1H, ArH; J = 7.6 Hz), 7.37 (d, 1H, ArH; J = 7.6 Hz), 7.55 (t, 1H, ArH; J = 7.6 Hz), 7.92 (d, 1H, ArH; J = 7.6 Hz), 12.43 (bs, 1H, NH; exchangeable with D2O), 14.54 (bs, 1H, NH; exchangeable with D2O); 13C-NMR (DMSO-d6) δ:113.5, 118.1, 122.3, 123.3, 132.3, 136.1, 143.5, 149.6, 179.5. Anal. Calcd for C9H6N4S (202.24): C, 53.45; H, 2.99; N, 27.70 Found: C, 53.60; H, 3.23; N, 27.58.

3.11. 5,10-Dihydro-[1,2,4]triazino[5,6-b]quinoxaline-3-thiol (23)

A mixture of quinoxaline-2,3(1H,4H)-dione (1''', 0.01 mol), thiosemicarbazide (0.011 mol) and anhydrous potassium carbonate (0.015 mol) [71] was stirred in water (500 mL) for 2 h at R.T., and then refluxed for 5 h. The mixture was cooled, filtered, and the filtrate was acidified with acetic acid. The solid mass obtained was separated by filtration, washed with water and dried. The product was recrystallized from ethanol. The product was obtained as off white crystals; yield 1.81 g, 83.6%; m.p. >300 °C; IR (KBr): 3160, 3440 cm−1 (NH); 1H-NMR (DMSO-d6) δ 7.03 (d, 2H, ArH; J = 7.6 Hz), 7.08 (t, 2H, ArH; J = 7.6 Hz), 11.88 (bs, 2H, 2NH; exchangeable with D2O), 14.50 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C9H7N5S (217.25): C, 49.76; H, 3.25; N, 32.24 Found: C, 49.90; H, 3.40; N, 32.00.

3.12. 3-Hydrazinyl-5H-[1,2,4]triazino[5,6-b]indole (24)

A mixture of 5H-[1,2,4]triazino[5,6-b] indole-3-thiol (22, 0.01 mol) [71] and hydrazine hydrate (10 mL, 98%) was heated on water bath for 5 h. The product was collected, washed with ethanol and dried. It was recrystallized from ethanol. The product was obtained as yellow crystals; yield 1.85 g, 92.6%; m.p. 260–261 °C; IR (KBr): 3176, 3242, 3300, 3404 cm−1 (NH, NH2); 1H-NMR (DMSO-d6) δ 4.31 (bs, 2H, NH2; exchangeable with D2O), 7.24 (t, 1H, ArH; J = 7.9 Hz), 7.38 (d, 1H, ArH; J = 7.9 Hz), 7.44 (t, 1H, ArH; J = 7.9 Hz), 8.07 (d, 1H, ArH; J = 7.9 Hz), 8.54 (bs, 1H, NH; exchangeable with D2O), 11.82 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C9H8N6 (200.20): C, 53.99; H, 4.03 N, 41.98 Found: C, 54.10; H, 4.22; N, 41.80.

3.13. 3-Hydrazinyl-5,10-dihydro-[1,2,4]triazino[5,6-b]quinoxaline (25)

A mixture of 5,10-dihydro-[1,2,4]triazino[5,6-b]quinoxaline-3-thiol (23, 0.01 mol) and hydrazine hydrate (10 mL, 98%) [71] was heated on water bath for 5 h. The product was collected, washed with ethanol, dried and recrystallized from ethanol. The product was obtained as buff crystals; yield 1.83 g, 85.2%; m.p. 255–256 °C; IR (KBr): 3176, 3237, 3271, 3300, 3402 cm−1 (NH, NH2); 1H-NMR (DMSO-d6) δ 4.55 (bs, 2H, NH2; exchangeable with D2O), 7.07 (t, 2H, ArH; J = 7.6 Hz), 7.29 (d, 2H, ArH; J = 7.6 Hz), 8.90 (bs, 1H, NH; exchangeable with D2O),11.89 (bs, 2H, 2 NH; exchangeable with D2O). Anal. Calcd for C9H9N7 (215.21): C, 50.23; H, 4.22; N, 45.56 Found: C, 50.40; H, 4.45; N, 45.39.

3.14. Ethyl [1,2,4]triazolo[3,4-c][1,2,4]triazino[5,6-b]-5H-indole-5-ethanoate (26)

A mixture of 3-hydrazinyl-5H-[1,2,4]triazino[5,6-b]indole (24, 0.01 mol) [70] and diethyl malonate (20 mL) [72] was heated 10 h. The product was filtered off, washed with ethanol and dried. It was recrystallized from ethanol. The product was obtained as buff crystals; yield 2.55g, 86.3%; m.p. 283–284 °C; IR (KBr): 1737 (C=O), 3408 cm−1 (NH); 1H-NMR (DMSO-d6) δ 1.16 (t, 3H, CH3-ester; J = 6.7 Hz), 4.12 (s, 2H, CH2), 4.31 (q, 2H, CH2-ester; J = 6.7 Hz), 7.20–7.26 (m, 1H, ArH), 7.36–7.38 (m, 1H, ArH), 7.60–7.65 (m, 1H, ArH), 8.00–8.10 (m, 1H, ArH), 12.18 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C14H12N6O2 (296.28): C, 56.75; H, 4.08; N, 28.36 Found: C, 56.90; H, 4.24; N, 28.21.

3.15. Ethyl [1,2,4]triazolo[3,4-c][1,2,4]triazino[5,6-b]-5,10-dihydroquinoxaline-5-ethanoate (27)

A mixture of 3-hydrazinyl-5,10-dihydro-[1,2,4]triazino[5,6-b]quinoxaline (25, 0.01 mol) and diethyl malonate (20 mL) [72] was heated 10 h. The product was filtered off, washed with ethanol, dried and recrystallized from ethanol. The product was obtained as brown crystals; yield 2.77 g, 89.0%; m.p. 280–281 °C; IR (KBr): 1713 (C=O), 3150, 3430 cm(NH); 1H-NMR (DMSO-d6) δ 1.09 (t, 3H, CH3; J = 6.9 Hz), 4.09 (q, 2H, CH2; J = 6.9 Hz), 4.66 (s, 2H, CH2), 7.24 (d, 1H, ArH; J = 8.4 Hz), 7.37–7.41 (m, 2H, ArH), 7.74 (d, 1H, ArH; J = 8.4 Hz), 12.10 (bs, 2H, 2 NH; exchangeable with D2O). 13C-NMR (DMSO-d6) δ: 14.4, 34.5, 62.0, 123.7, 128.5, 129.7, 145.2, 147.0, 152.1, 168.4. Anal. Calcd for C14H13N7O2 (311.30): C, 54.02; H, 4.21; N, 31.50 Found: C, 54.25; H, 4.41; N, 31.38.

3.16. [1,2,4]Triazolo[3,4-c][1,2,4]triazino[5,6-b]-5H-indole-5-ethanoic acid hydrazide (28)

A solution of ester 26 (0.01 mol) in ethanol (30 mL) and hydrazine hydrate 98% (10 mL) [72] was refluxed for 6 h. The product was collected, washed with ethanol and dried. It was recrystallized from ethanol. The product was obtained as buff crystals; yield 2.15 g, 76.5%; m.p. >300 °C, IR (KBr): 1657 (C=O), 3176, 3242, 3308, 3434 cm−1 (NH, NH2); 1H-NMR (DMSO-d6) δ 4.24 (s, 2H, CH2), 7.24–7.30 (m, 1H, ArH), 7.36–7.41 (m, 1H, ArH), 7.62–7.68 (m, 1H, ArH), 8.03–8.10 (m, 1H, ArH), 9.20 (bs, 1H, NH), 9.89 (bs, 2H, NH2; exchangeable with D2O), 12.18 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C12H10N8O (282.26): C, 51.06; H, 3.57; N, 39.70 Found: C, 51.20; H, 4.11; N, 39.48.

3.17. [1,2,4]Triazolo[3,4-c][1,2,4]triazino[5,6-b]-5,10-dihydroquinoxaline-5-ethanoic acid hydrazide (29)

A solution of ester 27 (0.01 mol) in ethanol (30 mL) and hydrazine hydrate 98% (10 mL) [72] was refluxed for 6 h. The product was collected, washed with ethanol, dried and recrystallized from ethanol. The product was obtained as orange crystals; yield 2.36 g, 79.6%; m.p. 250–251 °C; IR (KBr): 1675 (C=O), 3190, 3306, 3434 cm−( (NH, NH2); 1H-NMR (DMSO-d6) δ 4.34 (s, 2H, CH2), 7.24 (t, 1H, ArH; J = 7.6 Hz), 7.38 (d, 1H, ArH; J = 7.6 Hz), 7.44 (t, 1H, ArH; J = 7.6 Hz), 7.87 (d, 1H, ArH; J = 7.6 Hz), 8.90 (bs, 1H, NH; exchangeable with D2O), 9.61 (bs, 2H, NH2; exchangeable with D2O), 12.08 (bs, 2H, 2 NH; exchangeable with D2O). Anal. Calcd for C12H11N9O (297.28): C, 48.48; H, 3.73; N, 42.41 Found: C, 48.63; H, 3.90; N, 42.30.

3.18. [1,2,4]Triazolo[3,4-c][1,2,4]triazino[5,6-b]-5H-indole-5-N-(phenylcarbamothioyl) ethanoic acid hydrazide (30)

A mixture of acid hydrazide 28 (0.01 mol) and anhydrous potassium carbonate (0.05 mol) [71] in absolute ethanol (25 mL) was treated by drop wise addition of phenyl isothiocyanate (0.2 mol) in dry acetone (10 mL) .The reaction mixture was heated under reflux for 12 h. The product was collected, washed with ethanol and dried. The acetone was distilled under reduced pressure and the solid residue was dissolved in water, the product was isolated after acidification with 2 N HCl. The solid mass obtained was separated by filtration, washed with water, dried and recrystallized from ethanol The product was obtained as orange crystals; yield 3.34 g, 80.1%; m.p. 270–271 °C; IR (KBr): 1105 (C=S), 1651 (C=O), 3236, 3433 cm−1 (NH); 1H-NMR (DMSO-d6) δ 2.06 (s, 2H, CH2), 6.89–6.92 (m, 2H, ArH, NH), 7.24–7.28 (m, 4H, ArH), 7.50–7.53 (m, 4H, ArH), 9.88 (bs, 2H, NH; exchangeable with D2O), 11.82 (bs, 1H, NH; exchangeable with D2O); 13C-NMR (DMSO-d6) δ: 31.4, 117.5, 117.5, 117.6, 117.6, 121.7, 121.7, 122.1, 123.6, 127.2, 129.6, 129.7, 141.9, 155.0, 156.7, 178.7, 189.4. Anal. Calcd for C19H15N9OS (417.45): C, 54.67; H, 3.62; N, 30.20 Found: C, 54.40; H, 3.43; N, 30.42.

3.19. [1,2,4]Triazolo[3,4-c][1,2,4]triazino[5,6-b]-5,10-dihydroquinoxaline-5-N-(phenylcarbamothioyl)-ethanoic acid hydrazide (31)

A mixture of acid hydrazide 35 (0.001 mol) and anhydrous potassium carbonate (0.005 mol) in absolute ethanol (25 mL) was treated by dropwise addition of phenyl isothiocyanate (0.02 mol) [72] in dry acetone (10 mL) .The reaction mixture was heated under reflux for 12 h. The product was collected, washed with ethanol and dried. It was recrystallized from ethanol. The acetone was distilled under reduced pressure and the solid residue was dissolved in water, the product was isolated after acidification with 2 N HCl. The solid mass obtained was separated by filtration, washed with water, dried and recrystallized from ethanol. The product was obtained as buff crystals; yield 0.34 g, 80.0%; m.p. 275–276 °C; IR (KBr): 1109 (C=S), 1651 (C=O), 3236, 3433 cm−1 (NH); 1H-NMR (DMSO-d6) δ 4.25 (s, 2H, CH2), 7.00–7.54 (m, 5H, 4ArH, NH), 7.56–8.00 (m, 7H, 5ArH, 2NH), 12.05 (m, 2H, 2NH; exchangeable with D2O). Anal. Calcd for C19H16N10OS (432.46): C, 52.77; H, 3.73; N, 32.39 Found: C, 52.49; H, 3.53; N, 32.52.

3.20. 1-(5H-[1,2,4]Triazino[5,6-b]indol-3-yl)-3-methyl-1H-pyrazol-5(4H)-one (32)

A mixture of 3-hydrazinyl-5H-[1,2,4]triazino[5,6-b]indole (24, 0.01 mol) and ethyl acetoacetate (0.011 mol) [73] in acetic acid (20 mL) was stirred with refluxing for 4 h. The reaction mixture was evaporated till dryness, the product was collected, washed with ethanol and dried. It was recrystallized from ethanol. The product was obtained as buff crystals; yield 2.47 g, 93.0%; m.p. 330–331 °C; IR (KBr): 1712 (C=O), 3128, 3438 cm−1 (NH); 1H-NMR (DMSO-d6) δ 1.80 (s, 3H, CH3), 2.43 (s, 2H, CH2), 7.45 (t, 1H, ArH; J = 6.7 Hz), 7.61 (d, 1H, ArH; J = 6.7 Hz), 7.72 (t, 1H, ArH; J = 6.7 Hz), 8.36 (d, 1H, ArH; J = 6.7 Hz), 12.00 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C13H10N6O (266.26): C, 58.64; H, 3.79; N, 31.56 Found: C, 58.80; H, 3.90; N, 31.43.

3.21. 1-(5,10-Dihydro-[1,2,4]triazino[5,6-b]quinoxalin-3-yl)-3-methyl-1H-pyrazol-5(4H)-one (33)

A mixture of 3-hydrazinyl-5,10-dihydro-[1,2,4]triazino[5,6-b]quinoxaline 25 ( 0.01 mol) and ethyl acetoacetate (0.011 mol) [73] in acetic acid (20 mL) was stirred with refluxing for 4 h. Evaporation till dryness, the product was collected, washed with ethanol and dried. It was recrystallized from ethanol. The product was obtained as buff crystals; yield 2.45 g, 87.2%; m.p. 239–240 °C; IR (KBr): 1693 (C=O), 3161, 3438 cm−1 (NH); 1H-NMR (DMSO-d6) δ 2.29 (s, 3H, CH3), 2.92 (s, 2H, CH2), 7.23 (t, 1H, ArH; J = 7.6 Hz), 7.33 (d, 1H, ArH; J = 7.6 Hz), 7.39 (t, 1H, ArH; J = 7.6 Hz), 7.98 (d, 1H, ArH; J = 7.6 Hz), 10.99 (bs, 1H, NH; exchangeable with D2O), 11.68 (bs, 1H, NH; exchangeable with D2O). 13C-NMR (DMSO-d6) δ: 15.0, 43.8, 127.9, 130.7, 144.5, 152.9, 155.7, 161.8, 170.0. Anal. Calcd for C13H11N7O (281.27): C, 55.51; H, 3.94; N, 34.86 Found: C, 55.68; H, 4.03; N, 34.70.

3.22. 1-(5H-[1,2,4]Triazino[5,6-b]indol-3-yl)-3-methyl-4-(propan-2-ylidene)-1H-pyrazol-5(4H)-one (34)

A solution of 1-(5H-[1,2,4]triazino[5,6-b]indol-3-yl)-3-methyl-1H-pyrazol-5(4H)-one (32, 0.01 mol) in acetone (25 mL) [73] was stirred with refluxing for 20 h. The reaction mixture was evaporated till dryness, cooled. 20 mL of water was added in order to precipitate the product. The crude product was filtered, washed with ethanol, dried, and recrystallized from ethanol. The product was obtained as off white crystals; yield 2.47 g, 80.7%; m.p. 360–361 °C; IR (KBr): 1713 (C=O), 3439 cm−1 (NH); 1H-NMR (DMSO-d6) δ 1.87 (s, 6H, 2 CH3), 2.49 (s, 3H, CH3), 7.44 (t, 1H, ArH; J = 7.6 Hz), 7.62 (d, 1H, ArH; J = 7.6 Hz), 7.71 (t, 1H, ArH; J = 7.6 Hz), 8.36 (d, 1H, ArH; J = 7.6 Hz), 13.08 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C16H14N6O (306.32): C, 62.74; H, 4.61; N, 27.44 Found: C, 62.97; H, 5.03; N, 27.70.

3.23. 1-(5,10-Dihydro-[1,2,4]triazino[5,6-b]quinoxalin-3-yl)-3-methyl-4-(propan-2-ylidene)-1H-pyrazol-5(4H)-one (35)

A solution of 1-(5,10-dihyro-[1,2,4]triazino[5,6-b]quinoxalin-3-yl)-3-methyl-1H-pyrazol-5(4H)-one (33, 0.01 mol) in acetone (25 mL) [73] was stirred with refluxing for 20 h. Evaporation till dryness, cooled, water was added, filter the product was collected, washed with ethanol and dried. It was recrystallized from ethanol. The product was obtained as buff crystals; yield 2.66 g, 82.9%; crystals; m.p. 263–264 °C; IR (KBr): 1701 (C=O), 3202, 3433 cm−1 (NH); 1H-NMR (DMSO-d6) δ 2.30 (s, 3H, CH3), 2.42 (s, 3H, CH3), 2.93 (s, 3H, CH3), 7.00–7.05 (m, 2H, ArH), 7.35–7.39 (m, 2H, ArH), 11.88 (bs, 2H, 2 NH; exchangeable with D2O); 13C-NMR (DMSO-d6) δ:14.6, 19.2, 107.3, 115.6, 123.5, 126.1, 137.4, 153.0, 155.7, 159.5, 161.8. Anal. Calcd for C16H15N7O (321.34): C, 59.80; H, 4.71; N, 30.51 Found: C, 60.03; H, 4.93; N, 30.37.

3.24. 2-((2-(5H-[1,2,4]Triazino[5,6-b]indol-3-yl)hydrazono)methyl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one (36)

A mixture of 3-hydrazinyl-5H-[1,2,4]triazino[5,6-b]indole (24, 0.001 mol) and 6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indole-2-carbaldehyde (1', 0.0011 mol) was refluxed in ethanol (30 mL) for 6 h. The product was collected, washed with ethanol and dried, it was recrystallized from ethanol. The product was obtained as brown crystals; yield 0.32 g, 87.6%; m.p. 290–291 °C; IR (KBr): IR (KBr): 1652 (C=O), 3234, 3439 cm−1 (NH); 1H-NMR (DMSO-d6) δ 1.00 (s, 6H, 2 CH3), 2.21 (s, 2H, CH2), 2.68 (s, 2H, CH2), 6.53 (s, 1H, CH-pyrrole), 7.30 (t, 1H, ArH; J = 7.9 Hz), 7.46 (d, 1H, ArH; J = 7.9 Hz ), 7.51 (t, 1H, ArH; J = 7.9 Hz), 8.03 (s, 1H, CH=N), 8.13 (d, 1H, ArH; J = 7.9 Hz), 8.34 (bs, 1H, NH; exchangeable with D2O), 11.52 (bs, 1H, NH; exchangeable with D2O), 11.89 (bs, 1H, NH; exchangeable with D2O). 13C-NMR (DMSO-d6) δ: 28.7, 35.7, 36.6, 52.2, 108.2, 112.7, 119.4, 119.9, 120.6, 122.5, 129.4, 129.6, 136.1, 138.6, 140.3, 146.3, 148.8, 158.7, 192.6. Anal. Calcd for C20H19N7O (373.41): C, 64.33; H, 5.13; N, 26.26 Found: C, 64.63; H, 4.90; N, 26.30.

3.25. 2-((2-(5,10-Dihydro-[1,2,4]triazino[5,6-b]quinoxalin-3-yl)hydrazono)methyl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one (37)

A mixture of 3-hydrazinyl-5,10-dihydro-[1,2,4]triazino[5,6-b] quinoxaline (25, 0.001 mol) and 6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indole-2-carbaldehyde (1a, 0.0011 mol) was refluxed in ethanol (30 mL) for 6 h. The product was collected, washed with ethanol, dried and recrystallized from ethanol. The product was obtained as yellow crystals; yield 0.33 g, 86.6%; m.p. 155–156 °C; IR (KBr): 1651 (C=O), 3236, 3433 cm−1 (NH); 1H-NMR (DMSO-d6) δ 0.97 (s, 3H, CH3), 1.00 (s, 3H, CH3), 2.21 (s, 2H, CH2), 2.69 (s, 2H, CH2), 6.82 (s, 1H, CH-pyrrole), 7.03–7.06 (m, 2H, ArH), 7.07–7.10 (m, 2H, ArH), 9.45 (s, 1H, CH=N), 10.34 (bs, 1H, NH; exchangeable with D2O), 11.16 (bs, 1H, NH; exchangeable with D2O), 11.91 (bs, 1H, NH; exchangeable with D2O), 12.11 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C20H20N8O (388.43): C, 61.84; H, 5.19; N, 28.85 Found: C, 62.00; H, 5.31; N, 28.70.

3.26. 3-(5H-[1,2,4]Triazino[5,6-b]indol-3-yl)-2-(6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indol-2-yl)thiazolidin-4-one (38)

A mixture of 2-((2-(5H-[1,2,4]triazino[5,6-b]indol-3-yl)hydrazono)methyl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one (36, 0.001 mol) and thioglycolic acid (0.0012 mol) was refluxed in dry benzene (30 mL) on water bath for 10 h, cooled and poured onto water. The reaction mixture was extracted with benzene three times, washed with sodium bicarbonate, water, dried over anhydrous sodium sulfate, then concentrated to half its volume. The separated solid product was filtered, washed ethanol, dried and recrystallized from ethanol. The product was obtained as buff crystals; yield 0.33 g, 77.2%; m.p. 215–216 °C; IR (KBr): 1650, 1710 (C=O), 3427 cm−1 (NH); 1H-NMR (DMSO-d6) δ 1.00 (s, 6H, 2 CH3), 2.20 (s, 2H, CH2), 2.69 (s, 2H, CH2), 3.35–3.49 (m, 2H, CH2 –thiazol.), 6.52 (s, 1H, CH-pyrrole), 7.24 (t, 1H, ArH; J = 7.7 Hz), 7.43–7.49 (m, 2H, ArH), 7.93 (s, 1H, H-thiazol.), 8.08 (d, 1H, ArH; J = 7.7 Hz), 11.46 (bs, 1H, NH; exchangeable with D2O), 12.03 (bs, 1H, NH; exchangeable with D2O). Anal. Calcd for C22H20N6O2S (432.50): C, 61.10; H, 4.66; N, 19.43 Found: C, 61.36; H, 4.86; N, 19.21.

3.27. N'-2-((2-(5,10-Dihydro-[1,2,4]triazino[5,6-b]quinoxalin-3-yl)hydrazono)methyl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-ylidene)benzohydrazide (39)

To a solution of 2-((2-(5,10-dihydro-[1,2,4]triazino[5,6-b]quinoxalin-3-yl)hydrazono)methyl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one (37, 0.001 mol) in ethanol (30 mL) was added benzoyl hydrazine (0.163 g, 0.0012 mol) and two drops of acetic acid. The reaction mixture was heated under reflux for 6 h, partially concentrated and cooled. The separated solid product was filtered, washed with ethanol, dried and recrystallized from ethanol. The product was obtained as yellow crystals; yield 0.40 g, 79.7%; m.p. 175–176 °C; IR (KBr): 1644 (C=O), 3236, 3426 cm−1 (NH); 1H-NMR (DMSO-d6) δ 1.01 (s, 6H, 2 CH3), 2.22 (s, 2H, CH2), 2.69 (s, 2H, CH2), 6.66 (s, 1H, CH-pyrrole), 7.17 (bs, 1H, NH; exchangeable with D2O), 7.49–7.51 (m, 5H, ArH), 7.87–7.90 (m, 4H, ArH), 8.25 (s, 1H, CH=N), 10.56 (s, 1H, NH; exchangeable with D2O), 11.55 (s, 1H, NH; exchangeable with D2O), 11.63 (s, 1H, NH; exchangeable with D2O), 12.00 (s, 1H, NH; exchangeable with D2O); 13C-NMR (DMSO-d6) δ: 28.8, 29.0, 35.8, 36.7, 110.6, 112.3, 120.2, 128.2, 128.3, 128.3, 128.3, 129.2, 129.2, 132.3, 132.6, 140.2, 146.6, 148. 3, 158.4, 172.7. Anal. Calcd for C27H26N10O (506.56): C, 64.02; H, 5.17; N, 27.65 Found: C, 63.89; H, 4.92; N, 27.80.

3.28. 3-(6,6-Dimethyl-6,7-dihydro-1H-indol-4(5H)-one)-1,5-dihydro-[1,2,4]triazolo[3,4-c]-5,10-dihydro-[1,2,4]triazino[5,6-b]quinoxaline (40)

A mixture of 2-((2-(5,10-dihydro-[1,2,4]triazino[5,6-b]quinoxalin-3-yl)hydrazono)methyl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one (37, 0.001 mol) and acetic anhydride (15 mL) was heated on a boiling water bath for 10 h. The reaction mixture was poured onto crushed ice, the precipitated product was filtered, washed with water, dried and recrystallized from ethanol/chloroform. The product was obtained as brown crystals; yield 0.28 g, 73.5%; m.p. 280–281 °C; IR (KBr): 1662 (C=O), 3234, 3429 cm−1 (NH); 1H-NMR (DMSO-d6) δ 1.08 (s, 6H, 2 CH3), 2.32 (s, 2H, CH2), 2.75 (s, 2H, CH2), 6.79 (s, 1H, CH-pyrrole), 7.09–7.12 (m, 1H, ArH), 7.31 (d, 1H, ArH; J = 7.6 Hz), 7.38–7.42 (m, 2H, ArH), 12.23 (bs, 3H, 3NH; exchangeable with D2O). Anal. Calcd for C20H18N8O(386.41): C, 62.17; H, 4.70; N, 29.00 Found: C, 61.92; H, 4.62; N, 29.19.

3.29. 2-(1-(5H-[1,2,4]Triazino[5,6-b]indol-3-yl)-3-(4-bromophenyl)-1H-pyrazol-5-yl)-(6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one (41)

To a solution of 2-(3-(4-bromophenyl)-3-oxoprop-1-enyl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one (2, 0.001 mol) in ethanol (15 mL) was added 3-hydrazinyl-5H-[1,2,4]triazino[5,6-b]indole (24, 0.0012 mol) and acetic acid (5 mL). The reaction mixture was heated under reflux for 7 h, concentrated till dryness, poured onto H2O (50 mL), 10% bromine water (5 mL) was added, the mixture stirred overnight, and poured onto crushed ice. The separated solid product was filtered, washed with ethanol, dried and recrystallized from ethanol. The product was obtained as brown crystals; yield 0.44 g, 80.8%; m.p. 150–151 °C; IR (KBr): 1649 (C=O), 3439 cm−1 (NH); 1H-NMR (DMSO-d6) δ 1.01 (s, 6H, 2 CH3), 2.35 (s, 2H, CH2), 2.67 (s, 2H, CH2), 6.12 (s, 1H, CH-pyrazole), 6.70 (s, 1H, CH-pyrrole), 7.30–7.37 (m, 2H, ArH), 7.50–7.54 (m, 4H, ArH), 7.73 (d, 2H, ArH; J = 7.7 Hz), 11.35 (s, 1H, NH; exchangeable with D2O), 11.78 (s, 1H, NH; exchangeable with D2O); 13C-NMR (DMSO-d6) δ: 28.9, 33.2, 45.4, 57.8, 108.2, 111.3, 113.6, 120.8, 121.8, 122.6, 124.7, 124.9, 125.2, 125.7, 126.9, 127.6, 128.9, 132.3, 142.2, 146.6, 148.5, 150.4, 157.6, 190.8. Anal. Calcd for C28H22BrN7O (552.42): C, 60.88; H, 4.01; N, 17.75 Found: C, 61.00; H, 4.20; N, 17.58.

3.30. General Procedure for the Preparation of Compounds 42–44

To a solution of 2-(3-aryl-3-oxoprop-1-enyl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one 24 (0.001 mol) in ethanol (15 mL) was added 3-hydrazinyl-5,10-dihydro-[1,2,4]triazino[5,6-b] quinoxaline (25, 0.0012 mol) and acetic acid (5 mL). The reaction mixture was heated under reflux for 7 h, concentrated till dryness, poured onto H2O (50 mL), 10% bromine water (5 mL) was added, the mixture stirred overnight, and poured onto crushed ice. The separated solid product was filtered, washed with ethanol, dried and recrystallized from ethanol.

2-(3-(4-Bromophenyl)-1-(5,10-dihydro-[1,2,4]triazino[5,6-b]quinoxalin-3-yl)-1H-pyrazol-5-yl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one (42). Orange crystals; yield 0.49 g, 88.0%; m.p. 289–290 °C; IR (KBr): 1680 (C=O), 3232, 3454 cm−1 (NH); 1H-NMR (DMSO-d6) δ 1.01 (s, 6H, 2 CH3), 2.23 (s, 2H, CH2), 2.71 (s, 2H, CH2), 6.80 (s, 1H, CH-pyrazole), 6.91 (s, 1H, CH-pyrrole), 7.50–7.60 (m, 4H, ArH), 7.76 (d, 2H, ArH; J = 8.4 Hz), 7.92 (d, 2H, ArH; J = 8.4 Hz), 12.06 (m, 3H, 3NH; exchangeable with D2O); 13C-NMR (DMSO-d6) δ:.28.8, 34.5, 48.6, 49.7, 51.5, 99.1, 115.6, 122.4, 124, 125.8, 129.0, 129.8, 131.8, 138.0, 140.1, 1418, 143.9, 146.6, 151.7, 155.6, 193.3. Anal. Calcd for C28H23BrN8O (567.44): C, 59.27; H, 4.09; N, 19.75 Found: C, 59.15; H, 4.21; N, 19.50.

2-(3-(4-Chlorophenyl)-1-(5,10-Dihydro-[1,2,4]triazino[5,6-b]quinoxalin-3-yl)-1H-pyrazol-5-yl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one (43). Brown crystals; yield 0.48 g, 82.3%; m.p. 229–230 °C; IR (KBr): 1681 (C=O), 3250, 3405 cm−1 (NH); 1H-NMR (DMSO-d6) δ 1.00 (s, 6H, 2 CH3), 2.23 (s, 2H, CH2), 2.71 (s, 2H, CH2), 6.90 (s, 1H, CH-pyrazole), 7.27 (s, 1H, CH-pyrrole), 7.50–7.55 (m, 2H, ArH), 7.69–7.72 (m, 4H, ArH), 8.00–8.12 (m, 2H, ArH), 12.18 (bs, 3H, 3NH; exchangeable with D2O); 13C-NMR (DMSO-d6) δ:.28.8, 39.8, 40.2, 40.6, 54.2, 64.1, 109.6, 115.8, 126.3, 129.5, 129.6, 129.6, 130.6 130.8, 137.4, 143.2, 143.7, 150.2, 155.9, 169.5, 192.8. Anal. Calcd for C28H23ClN8O (522.99): C, 64.30; H, 4.43; N, 21.43 Found: C, 64.26; H, 4.20; N, 21.66.

2-(1-(5,10-Dihydro-[1,2,4]triazino[5,6-b]quinoxalin-3-yl)-3-(4-methoxyphenyl)-1H-pyrazol-5-yl)-6,6-dimethyl-6,7-dihydro-1H-indol-4(5H)-one (44). Brown crystals; yield 0.45 g, 86.8%; m.p. 280–281 °C; IR (KBr): 1681 (C=O), 3151, 3436 cm−1 (NH); 1H-NMR (DMSO-d6) δ 1.02 (s, 6H, 2 CH3), 2.24 (s, 2H, CH2), 2.72 (s, 2H, CH2), 2.94 (s, 3H, OCH3), 6.75–6.78 (m, 1H, CH-pyrazole), 6.85 (s, 1H, CH-pyrrole), 7.06 (d, 2H, ArH; J = 8.4 Hz), 7.27–7.36 (m, 2H, ArH), 7.40 (d, 2H, ArH; J = 8.4 Hz), 8.01–8.09 (m, 2H, ArH), 11.95 (bs, 2H, 2NH; exchangeable with D2O), 12.05 (bs, 1H, NH; exchangeable with D2O). 13C-NMR (DMSO-d6) δ: 28.6, 35.0, 43.1, 53.8, 56.1, 100.1, 115.6, 117.1, 117.3, 121.5, 123.5, 123.8, 126.1, 129.5, 135.2, 140.1, 144.7, 149.7, 152.3, 155.7, 160.8, 193.0. Anal. Calcd for C29H26N8O2 (518.57): C, 67.17; H, 5.05; N, 21.61 Found: C, 67.30; H, 5.18; N, 21.41.

3.31. Biological Activity Assay

3.31.1. Inhibition Zone Measurement (IZ)

Compounds 244 were evaluated in vitro for antimicrobial activity against the following four organisms: Escherichia coli ATCC8739, Pseudomonas aeruginosa ATCC 9027 as examples of Gram-negative bacteria, Staphylococcus aureus ATCC 6583P as an example of Gram-positive bacteria, and Candida albicans ATCC 2091 as an example of a yeast-like fungus have been studied by using the Nutrient Agar (NA) and Sabouraud Dextrose Agar (SDA) diffusion methods [74], respectively, in N,N-dmethylformamide as solvent. The bacteria were subcultured on Nutrient Agar medium (NA), whereas, fungi were subcultured on Sabouraud Dextrose Agar (SDA). Petri plates (150 mm × 15 mm) were prepared by pouring 60 mL of NA or SDA and allowing it to solidify. Plates were dried and 1 mL of each standardized inoculums suspension was poured and uniformly spread. The excess inoculums was drained and the inoculums was allowed to dry for 15 min. Eight equidistant wells were made in the medium using a sterile cork borer (6 mm in diameter and 75 μL of the test chemicals (1 mg/mL) diluted in DMF were placed into the wells. The plates containing bacterial and fungi species were incubated at 37 °C for 24 h. The tests were carried in triplicate. Ampicillin trihydrate (10.0 µg/disc), ciprofloxacin (5.0 µg/disc), impenam (10.0 µg/disc), and clotrimazole (100.0 µg/disc) were used as standard antibacterial and antifungal agents, respectively. (DMF) alone showed no inhibition zone. The plates were incubated at 37 °C for 24 h. The results were recorded for each tested compound as the average diameter of inhibition zones of bacterial growth around the disks in mm.

3.31.2. Minimal inhibitory concentration (MIC)

MIC measurements [75] were carried out for compounds that showed significant inhibition zones using the twofold serial dilution technique. The compounds 244 were prepared in a concentration range of 200, 100, 50, 25, and 12.5 µg/mL. The microdilution susceptibility test in Muller-Hinton broth (oxoid) and Sabouraud Liquid Medium (oxoid) were used for the determination of antibacterial and antifungal activity. The microorganism suspensions at 106 CFU/mL (colony forming unit/mL) were used to inoculate the prepared test compounds in the above mentioned serial dilution broth. The culture tubes were incubated at 37 °C for 24–48 h. At the end of the incubation period the growth of bacteria was observed by turbidity measurements [75]. The MIC is defined as the lowest concentration that showed no bacterial growth.

4. Conclusions

The objective of the present study was to synthesize and investigate the antimicrobial and antifungal activity of a new series of pyrazolines and pyrazoles in the hope of discovering new structural leads serving as antimicrobial agents. Some new pyrazoline and pyrazole derivatives have been prepared, and their physical properties were characterized. The biological activity of the compounds 244 was evaluated by the agar diffusion method against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aurous and Candida albicans. None of the investigated compounds not showed any activity against the test organisms Escherichia coli and Pseudomonas aeruginosa. Compound 16 has good antimicrobial activity against Staphylococcus aureus, comparable to that of ampicillin and ciprofloxacin, while compound 17 has remarkable antimicrobial activity against Staphylococcus aureus, exceeding that of ampicillin, ciprofloxacin and imipenam. In addition, compound 17 has comparable IZ against Candida albicans comparable to that of clotrimazole. On the other hand, the minimal inhibitory concentration (MIC) of compounds 19, 20 and 31 against Candida albicans indicate good antifungal activity, comparable to that of clotrimazole. Based on the preliminary results, it can be seen that all four compounds 16, 17, 19 and 20 showing good antimicrobial and antifungal activity have benzenesulfonamide substituents as a common structural feature.

Acknowledgments

Thanks are due to the Pharmaceutical Chemistry Department, Faculty of Pharmacy, Alexandria University for the antimicrobial measurements.

Supplementary Materials

Supplementary materials can be accessed at: http://www.mdpi.com/1420-3049/18/3/2683/s1.

References

  1. Vogel, S.; Ohmayer, S.; Brunner, G.; Heilmann, J. Natural and nonnatural prenylated chalcones: Synthesis, cytotoxicity and antioxidative activity. Bioorg. Med. Chem. 2008, 16, 4286–4293, doi:10.1016/j.bmc.2008.02.079.
  2. Babasaheb, P.B.; Sachin, A.P.; Rajesh, N.G.; Balaji, L.K.; Balwant, S.H.; Santosh, N.K.; Shivkumar, S.J. Synthesis and biological evaluation of nitrogen-containing chalcones as possible anti-inflammatory and antioxidant agents. Bioorg. Med. Chem. Lett. 2010, 20, 730–733.
  3. Avila, H.P.; Smania, E.; Monache, F.D.; Smania, A. Structure-activity relationship of antibacterial chalcones. Bioorg. Med. Chem. 2008, 16, 9790–9794, doi:10.1016/j.bmc.2008.09.064.
  4. Suryawanshi, S.N.; Chandra, N.; Kumar, P.; Porwal, J.; Gupta, S. Chemotherapy of leishmaniasis part-VIII: Synthesis and bioevaluation of novel chalcones. Eur. J. Med. Chem. 2008, 43, 2473–2478, doi:10.1016/j.ejmech.2007.12.014.
  5. Lawrence, N.J.; Patterson, R.P.; Ooi, L.L.; Cook, D.; Ducki, S. Effects of α-substitutions on structure and biological activity of anticancer chalcones. Bioorg. Med. Chem. Lett. 2006, 16, 5844–5848, doi:10.1016/j.bmcl.2006.08.065.
  6. Mojzis, J.; Varinska, L.; Mojzisova, G.; Kostova, I.; Mirossay, L. Antiangiogenic effects of flavonoids and chalcones. Pharmacol. Res. 2008, 57, 259–265, doi:10.1016/j.phrs.2008.02.005.
  7. Cheng, J.H.; Hung, C.F.; Yang, S.C.; Wang, J.P.; Won, S.J.; Lin, C.N. Synthesis And Cytotoxic, Anti-Inflammatory, and Anti-Oxidant Activities Of 2',5'-Dialkoxyl chalcones As Cancer Chemopreventive Agents. Bioorg. Med. Chem. 2008, 16, 7270–7276, doi:10.1016/j.bmc.2008.06.031.
  8. Lahtchev, K.L.; Batovska, D.I.; Parushev, S.P.; Ubiyvovk, V.M.; Sibirny, A.A. Antifungal activity of chalcones: A mechanistic study using various yeast strains. Eur. J. Med. Chem. 2008, 43, 2220–2228, doi:10.1016/j.ejmech.2007.12.027.
  9. Liu, M.; Wilairat, P.; Cropft, S.L.; Tan, A.L.C.; Go, M.L. Structure-activity relationships of antileishmanial and antimalarial chalcones. Bioorg. Med. Chem. 2003, 11, 2729–2738, doi:10.1016/S0968-0896(03)00233-5.
  10. Go, M.L.; Wu, X.; Liu, X.L. Chalcones: An update on cytotoxic and chemoprotective properties. Curr. Med. Chem. 2005, 12, 483–499, doi:10.2174/0929867053363153.
  11. Liu, M.; Wilairat, P.; Go, M.L. Antimalarial alkoxylated and hydroxylated chalcones: Structure activity relationship analysis. J. Med. Chem. 2001, 44, 4443–4452, doi:10.1021/jm0101747.
  12. Bhat, B.A.; Dhar, K.L.; Puri, S.C.; Saxena, A.K.; Shanmugavel, M.; Qazi, G.N. Synthesis and biological evaluation of chalcones and their derived pyrazoles as potential cytotoxic agents. Bioorg. Med. Chem. Lett. 2005, 15, 3177–3180, doi:10.1016/j.bmcl.2005.03.121.
  13. Bandgar, B.P.; Gawande, S.S.; Bodade, R.G.; Gawande, N.M.; Khobragade, C.N. Synthesis and biological evaluation of a novel series of pyrazole chalcones as anti-inflammatory, antioxidant and antimicrobial agents. Bioorg. Med. Chem. 2009, 17, 8168–8173, doi:10.1016/j.bmc.2009.10.035.
  14. Zeba, N.S.; Mohammed, M.T.N.; Anis, A.; Asad, U.K. Thermal solvent-free synthesis of novel pyrazolyl chalcones and pyrazolines as potential antimicrobial agents. Bioorg. Med. Chem. Lett. 2011, 21, 2860–2865, doi:10.1016/j.bmcl.2011.03.080.
  15. Baraldi, P.G.; Manfredini, S.; Romagnoli, R.; Stevanato, L.; Zaid, A.N.; Manservigi, R. Synthesis andAnti-HSV-1 Activity of 6 Substituted Pyrazolo[3,4-d]Pyridazin-7-one Nucleosides’. NucleosidesNucleotides Nucleic Acids 1998, 17, 2165–2173, doi:10.1080/07328319808004307.
  16. Baraldi, P.G.; Bovero, A.; Fruttarolo, F.; Romagnoli, R.; Tabrizi, M.A.; Preti, D.; Varani, K.; Borea, P.A.; Moorman, A.R. New Strategies for the Synthesis of A3 Adenosine Receptor Antagonists. Bioorg. Med. Chem. 2003, 11, 4161–4169, doi:10.1016/S0968-0896(03)00484-X.
  17. Zitouni, G.T.; Chevallet, P.; Kiliç, F.S.; Erol, K. Synthesis of some thiazolyl-pyrazoline derivatives and preliminary investigation of their hypotensive activity. Eur. J. Med. Chem. 2000, 35, 635–641, doi:10.1016/S0223-5234(00)00152-5.
  18. Palaska, E.; Aytemir, M.; Uzbay, I.T.; Erol, D. Synthesis and antidepressant activities of some 3,5-diphenyl-2-pyrazolines. Eur. J. Med. Chem. 2001, 36, 539–543, doi:10.1016/S0223-5234(01)01243-0.
  19. Johnson, M.; Younglove, B.; Lee, L.; LeBlanc, R.; Holt, H.; Hills, P.; Mackay, H.; Brown, T.; Mooberry, L.S.; Lee, M. Design, synthesis, and biological testing of pyrazoline derivatives of combretastatin-A4. Bioorg. Med. Chem. Lett. 2007, 17, 5897–5901.
  20. Ratkovic, Z.; Juranic, Z.D.; Stanojkovic, T.; Manojlovic, D.; Vukicevic, R.D.; Radulovic, N.; Joksović, M.D. Synthesis, characterization, electrochemical studies and antitumor activity of some new chalcone analogues containing ferrocenyl pyrazole moiety. Bioorg. Chem. 2010, 38, 26–32, doi:10.1016/j.bioorg.2009.09.003.
  21. Rafia, B.; Syed, O.; Shafiya, Y.; Hinna, H.; Alam, M.S.; Mohammad, S.; Surender, S.; Kalim, J. Synthesis of some new 1,3,5-trisubstituted pyrazolines bearing benzene sulfonamide as anticancer and anti-inflammatory agents. Bioorg. Med. Chem. Lett. 2011, 21, 4301–4305.
  22. Sameena, B.; Kalim, J.; Shamim, A.; Rathish, I.G.; Surender, S.; Alam, M.S. Synthesis and biological evaluation of some new 2-pyrazolines bearing benzene sulfonamide moiety as potential anti-inflammatory and anti-cancer agents. Eur. J. Med. Chem. 2011, 46, 5763–5768, doi:10.1016/j.ejmech.2011.08.015.
  23. Rathish, I.G.; Kalim, J.; Shamim, A.; Sameena, B.; Alam, M.S.; Pillai, K.K.; Surender, S.; Vivek, B. Synthesis and antiinflammatory activity of some new 1,3,5-trisubstituted pyrazolines bearing benzene sulfonamide. Bioorg. Med. Chem. Lett. 2009, 19, 255–258.
  24. Joshi, S.; Mandhane, P.G.; Diwakar, S.D.; Dabhade, S.K.; Gill, C.H. Synthesis, analgesic and anti-inflammatory activities of some novel pyrazolines derivatives. Bioorg. Med. Chem. Lett. 2010, 20, 3721–3725, doi:10.1016/j.bmcl.2010.04.082.
  25. Mohammad, A.; Harish, K.; Suroor, A.K. Synthesis and pharmacological evaluation of pyrazoline derivatives as new anti inflammatory and analgesic agents. Bioorg. Med. Chem. Lett. 2008, 18, 918–922, doi:10.1016/j.bmcl.2007.12.043.
  26. Pawan, K.S.; Satish, K.; Pawan, K.; Pawan, K.; Dhirender, K.; Yogita, D.; Kamal, R.A. Synthesis and biological evaluation of some pyrazolylpyrazolines as anti-inflammatory-antimicrobial agents. Eur. J. Med. Chem. 2010, 45, 2650–2655, doi:10.1016/j.ejmech.2010.01.059.
  27. Suresh, K.; Veeresh, M.; Prashant, A.; Mahesh, P.; Pradeep, K.R.; Shivalingarao, M.; Thippeswamy, A.H.M.; Satyanarayana, D. Synthesis and pharmacological evaluation of a novel series of 5-(substituted) aryl-3-(3-coumarinyl)-1-phenyl-2-pyrazolines as novel anti-inflammatory and analgesic agents. Eur. J. Med. Chem. 2009, 44, 1682–1688, doi:10.1016/j.ejmech.2008.09.020.
  28. Amir, M.; Kumar, H.; Khan, S.A. Synthesis and pharmacological evaluation of pyrazoline derivatives as new anti-inflammatory and analgesic agents. Bioorg. Med. Chem. Lett. 2008, 18, 918–922, doi:10.1016/j.bmcl.2007.12.043.
  29. Rathish, I.G.; Javed, K.; Ahmad, S.; Bano, S. Synthesis, Antiinflammatory activity of some new 1,3,5-trisubstituted pyrazolines bearing benzene sulfonamide. Bioorg. Med. Chem. Lett. 2009, 19, 255–258, doi:10.1016/j.bmcl.2008.10.105.
  30. Barsoum, F.F.; Girgis, A.S. Facile synthesis of bis(4,5-dihydro-1H-pyrazole-1-carboxamides) and their thio-analogues of potential PGE2 inhibitory properties. Eur. J. Med. Chem. 2009, 44, 2172–2177, doi:10.1016/j.ejmech.2008.10.020.
  31. Khode, S.; Maddi, V.; Aragade, P.; Palkar, M.; Ronad, P.K.; Mamledesai, S.; Thippeswamy, A.H.M.; Satyanarayana, D. Synthesis and pharmacological evaluation of a novel series of 5-(substituted)aryl-3-(3-coumarinyl)-1-phenyl-2-pyrazolines as novel anti-inflammatory and analgesic agents. Eur. J. Med. Chem. 2009, 44, 1682–1688.
  32. Shoman, M.E.; Abdel-Aziz, M.; Aly, O.M.; Farag, H.H.; Morsy, M.A. Synthesis and investigation of anti-inflammatory activity and gastric ulcerogenicity of novel nitric oxide-donating pyrazoline derivatives. Eur. J. Med. Chem. 2009, 44, 3068–3076, doi:10.1016/j.ejmech.2008.07.008.
  33. Bhat, A.R.; Athar, F.; Azam, A. New Derivatives of 3,5-substituted-1,4,2-dioxazoles; Synthesis and Activity against Entamoeba histolytica. Eur. J. Med. Chem. 2009, 44, 926–936, doi:10.1016/j.ejmech.2008.02.001.
  34. Zuhal Ozdemir, H.; Burak Kandilci, B.G.X.; Unsal, C.; alısx, A.; Altan, B. Synthesis and studies on antidepressant and anticonvulsant activities of some 3-(2-furyl)-pyrazoline derivatives. Eur. J. Med. Chem. 2007, 42, 373–379, doi:10.1016/j.ejmech.2006.09.006.
  35. Manna, K.; Agrawal, Y.K. Microwave assisted synthesis of new indophenazine 1,3,5-trisubstruted pyrazoline derivatives of benzofuran and their antimicrobial activity. Bioorg. Med. Chem. Lett. 2009, 19, 2688–2692, doi:10.1016/j.bmcl.2009.03.161.
  36. Abdel-Wahab, B.F.; Abdel-Aziz, H.A.; Ahmed, E.M. Synthesis and antimicrobial evaluation of 1-(benzofuran-2-yl)-4-nitro-3-arylbutan-1-ones and 3-(benzofuran-2 yl)-4,5-dihydro-5-aryl-1-[4-(aryl)-1,3-thiazol-2-yl]-1H-pyrazoles. Eur. J. Med. Chem. 2009, 44, 2632–2635, doi:10.1016/j.ejmech.2008.09.029.
  37. El-Sayed, W.A.; Nassar, I.F.; Abdel-Rahman, A.A.-H. C-Furyl glycosides, II: Synthesis and antimicrobial evaluation of C-furyl glycosides bearing pyrazolines, isoxazolines, and 5,6-dihydropyrimidine-2(1H)-thione. Monatsh. Chem. 2009, 140, 365–370, doi:10.1007/s00706-008-0033-2.
  38. Jadhav, S.B.; Shastri, R.A.; Gaikwad, K.V.; Gaikwad, S.V. Synthesis and antimicrobial studies of some novel pyrazoline and isoxazoline derivatives. Eur. J. Chem. 2009, 6, S183–S188.
  39. Sakthinathan, S.P.; Vanangamudi, G.; Thirunarayanan, G. Synthesis, spectral studies and antimicrobial activities of some 2-naphthyl pyrazoline derivatives. Spectrochim. Acta A 2012, 95, 693–700, doi:10.1016/j.saa.2012.04.082.
  40. Kaplancikli, Z.A.; Turan-Zitouni, G.; Ozdemir, A.; Can, O.D.; Chevallet, P. Synthesis and antinociceptive activities of some pyrazoline derivatives. Eur. J. Med. Chem. 2009, 44, 2606–2610, doi:10.1016/j.ejmech.2008.09.002.
  41. Badri, N.; Acharya, D.S.; Mugdha, T.; Asish, K.S.; Ramarao, G.; Saroj, B.; Mahabir, P.K. Synthesis and antimalarial evaluation of 1,3,5-trisubstituted pyrazolines. Eur. J. Med. Chem. 2010, 45, 430–438, doi:10.1016/j.ejmech.2009.10.023.
  42. Carrion, M.D.; Luisa, C.; Lopez, L.C.; Camacho, M.E.; Tapias, V.; Escames, G.; Castroviejo, D.A.; Espinosa, A.; Gallo, M.A.; Entrena, A. Pyrazoles and pyrazolines as neural and inducible nitric oxide synthase (nNOS and iNOS) potential inhibitors (III). Eur. J. Med. Chem. 2008, 43, 2579–2591, doi:10.1016/j.ejmech.2008.01.014.
  43. Gokhan-Kelekc, N.; Koyunoglu, I.S.; Yabanoglu, S. New pyrazoline bearing 4(3H)-quinazolinone inhibitors of monoamine oxidase: Synthesis, biological evaluation, and structural determinants of MAO-A and MAO-B selectivity. Bioorg. Med. Chem. 2009, 17, 675–689, doi:10.1016/j.bmc.2008.11.068.
  44. Can, O.D.; Ozkay, U.D.; Kaplancikli, Z.A.; Ozturk, Y. Effects of some 1,3,5-trisubstitued-2-pyrazoline derivatives on depression and anxiety parameters of mice. Arch. Pharm. Res. 2009, 32, 1293–1299, doi:10.1007/s12272-009-1915-5.
  45. Havrylyuk, D.; Zimenkovsky, B.; Vasylenko, O.; Zaprutko, L.; Lesyk, R. Synthesis of novel thiazolone-based compounds containing pyrazoline moiety and evaluation of their anticancer activity. Eur. J. Med. Chem. 2009, 44, 1396–1404, doi:10.1016/j.ejmech.2008.09.032.
  46. Braulio, I.; Alexis, T.; Fabian, O.; Jairo, Q.; Rodrigo, A.; Manuel, N.; Adolfo, S.; Justo, C. Synthesis of novel pyrazolic analogues of chalcones and their 3-aryl-4-(3-aryl-4,5-dihydro-1H-pyrazol-5-yl)-1-phenyl-1H-pyrazole derivatives as potential antitumor agents. Bioorg. Med. Chem. 2010, 18, 4965–4974.
  47. Shrey, P.; Dhairya, B.; Mahesh, S.; Shailesh, T.; Abhay, B.; Manisha, P.; Hardevsinh, V.; Ashish, R.; Nilay, P.; Juliana, S.; et al. Snthesis of some novel benzofuran-2-yl(4,5-dihyro-3,5-substituted diphenylpyrazol-1-yl) methanones and studies on the antiproliferative effects and reversal of multidrug resistance of human MDR1-gene transfected mouse lymphoma cells in vitro. Eur. J. Med. Chem. 2011, 46, 1942–1948, doi:10.1016/j.ejmech.2011.02.045.
  48. Kuntal, M.; Yadvendra, K.A. Microwave assisted synthesis of new indophenazine 1,3,5-trisubstruted pyrazoline derivatives of benzofuran and their antimicrobial activity. J. Bioorg. Med. Chem. Lett. 2009, 19, 2688–2692, doi:10.1016/j.bmcl.2009.03.161.
  49. Chandra, T.; Garg, N.; Lata, S.; Saxena, K.K.; Kumar, A. Synthesis of substituted acridinyl pyrazoline derivatives and their evaluation for anti-inflammatory activity. Eur. J. Med. Chem. 2010, 45, 1772–1776.
  50. Jeong, T.-S.; Kim, K.S.; Kim, J.-R.; Cho, K.-Y.; Lee, S.; Lee, W.S. Novel 3,5-diaryl pyrazolines and pyrazole as low-density lipoprotein (LDL) oxidation inhibitor. Bioorg. Med. Chem. Lett. 2004, 14, 2719–2723, doi:10.1016/j.bmcl.2004.03.072.
  51. Abid, M.; Bhat, A.R.; Athar, F.; Azam, A. Synthesis, spectral studies and antiamoebic activity of new 1-N-substituted thiocarbamoyl-3-phenyl-2-pyrazolines. Eur. J. Med. Chem. 2009, 44, 417–425, doi:10.1016/j.ejmech.2007.10.032.
  52. Budakoti, A.; Bhat, A.R.; Azam, A. Synthesis of new 2-(5-substituted-3-phenyl- 2-pyrazolinyl)-1,3-thiazolino[5,4-b]quinoxaline derivatives and evaluation of their antiamoebic activity. Eur. J. Med. Chem. 2009, 44, 1317–1325, doi:10.1016/j.ejmech.2008.02.002.
  53. Faisal, H.; Attar, S.; Sadiq, U.; Amir, A. Synthesis, characterization, antiamoebic activity and cytotoxicity of novel series of pyrazoline derivatives bearing quinoline tail. Eur. J. Med. Chem. 2010, 45, 4669–4675, doi:10.1016/j.ejmech.2010.07.028.
  54. Bekhit, A.A.; Abdel-Aziem, T. Design, synthesis and biological evaluation of some pyrazole derivatives as anti-inflammatory-antimicrobial agents. Bioorg. Med. Chem. 2004, 12, 1935–1945, doi:10.1016/j.bmc.2004.01.037.
  55. Imran, A.; Waseem, A.W.; Amber, K.; Ashanul, H.; Aijaz, A.; Kishwar, S.; Nikhat, M. Synthesis and synergistic antifungal activities of a pyrazoline based ligand and its copper(II) and nickel(II) complexes with conventional antifungals. Microb. Pathog. 2012, 53, 66–73, doi:10.1016/j.micpath.2012.04.005.
  56. Kaplancikli, A.; Turan-Zitouni, G.; Ozdemir, A.; Can, O.D.; Chevallet, P. Synthesis and antinociceptive activities of some pyrazoline derivatives. Eur. J. Med. Chem. 2009, 44, 2606–2610, doi:10.1016/j.ejmech.2008.09.002.
  57. Maria, G.M.; Daniele, Z.; Valeria, F.; Luciano, V.; Elena, B. Synthesis and antimycobacterial activity of 5-aryl-1-isonicotinoyl-3-(pyridin-2-yl)-4,5-dihydro-1H-pyrazolederivatives. Farmaco 2001, 56, 593–599, doi:10.1016/S0014-827X(01)01098-9.
  58. Habibullah, K.; Shamshir, K.; Mohamed, J.A.; Bahar, A. Synthesis and antihepatotoxic activity of 5-(2,3-dihydro-1,4-benzodioxane-6-yl)-3-substituted-phenyl-4,5-dihydro-1H-pyrazole derivatives. Bioorg. Med. Chem. Lett. 2011, 21, 7251–7254, doi:10.1016/j.bmcl.2011.10.056.
  59. Berghot, M.A.; Moawad, E.B. Convergent synthesis and antibacterial activity of pyrazole and pyrazoline derivatives of diazepam. Eur. J. Pharm. Sci. 2003, 20, 173–179, doi:10.1016/S0928-0987(03)00162-3.
  60. Geeta, J.N.P.; Pramod, S.; Rawatb, B.S.; Rawata, M.S.M.; Joshi, G.C. Synthesis, characterization and fluorescence studies of 3,5-diaryl substituted 2-pyrazolines. Spectrochim. Acta A 2011, 78, 1075–1079.
  61. El-Sadek, M.M.; Faidalla, H.M.; El Soccary, N.N.; Hassan, S.Y. Synthesis And Spectral Studies Of Some New Pyrazolines And Pyrazoles. Egypt. J. Chem. 1995, 38, 403–418.
  62. Faidallah, H.M.; El-Sadek, M.M.; El-Massry, A.M.I.; Hassan, S.Y. Trisubstituted Pyrazoles of Possible Hypoglycemic Activity. Pak. J. Sci. Ind. Res. 1992, 35, 8–13.
  63. Faidallah, H.M.; Makki, M.S.I.; El-Massry, A.H.I.; Hassan, S.Y. Synthesis And Reactions of Some Ethyl 3-Aroyl-4-Aryl- 2-Pyrazoline-5-Carboxylates. Pharmazie 1997, 5, 101–105.
  64. Basaif, S.A.; Faidallah, H.M.; Hassan, S.Y. Synthesis and Biological Activity Of New Pyrazolines And Pyrazoles. Indian J. Heterocycl. Chem. 1996, 6, 53–58.
  65. Hassan, S.Y.; Basief, S.A.; Faidallah, H.M. Synthesis Of Noval Indan [1,2-c]Pyrazole-benzenesulfonylureas, Thioureas and Their Cyclized Derivatives. Egypt. J. Chem. 1999, 42, 213–220.
  66. Hassan, S.Y. Synthesis and Biological Activity of Some New Pyrazoline and Pyrimidine Derivatives. Braz. Chem. Soc. 2011, 22, 1286–1298, doi:10.1590/S0103-50532011000700014.
  67. González, F.G.; Guillen, M.G.; Perez, J.A.G.; Román, E.G. Reaction of 2-Amino-2-Deoxyheptoses With Cyclic Beta-Dicarbonyl Compounds. Carbohydr. Res. 1980, 78, 17–23, doi:10.1016/S0008-6215(00)83656-3.
  68. Bharat, K.; Vishal, P.; Sushma, R.; Rani, K.; Tewari, I.C. Synthesis and antimicrobial activity of some bromo-benzothiazolo pyrazolines. Int. J. Microbiol. Res. 2009, 1, 20–22.
  69. Kgokong, J.L.; Breytenbach, J.C. Synthesis of novel trifluoromethylquinoline and bis (trifluoromethyl)quinoline derivatives. S. Afr. J. Chem. S. 2000, 53, 100–103.
  70. Joseph, L.K.; Peter, P.S.; Gilbert, M.M. 1,2,4-triazino-[5,6-b]indole derivatives: Effects of the trifluoromethyl group on in vitro antimalarial activity. Bioorg. Med. Chem. 2005, 13, 2935–2942, doi:10.1016/j.bmc.2005.02.017.
  71. Mohamed, S.K.Y.; Ferial, M.A.; Khairy, M.H.; Mohamed, S.A. Synthesis and some reactions of 7-methyl-5-phenyl-5H-pyrazolo[3,4-e]1,2,4-triazine-3-thiol. J. Heterocycl. Chem. 1984, 21, 923–926, doi:10.1002/jhet.5570210362.
  72. Mohsen, A.M.E; Omar, S; El-Din, A.S.; Ibrahim, M.L.; El-Tombary, A.A. Synthesis and evaluation for antimicrobial and antihistaminic properties of new thiosemicarbazide and triazole derivatives of triazolo[4,3-a]quinazolin-5(4H)-ones. Alex. J. Pharm. Sci. 1991, 5, 213–215.
  73. Jack, D.R.; Deborah, A.C.; Wilmer, S.A.; Patick, J.S. Synthesis and reactions of 4-isipropylidene-1-aryl-3-methyl-2-pyrazolin-5-ones. J. Heterocycl. Chem. 1987, 24, 149–153, doi:10.1002/jhet.5570240128.
  74. Abou-Elela, G.M.; Ibrahim, H.A.; El-Helow, E.; Sabry, S. Abundance and Antagonistic Interactions among Bacterioplankton in Suez Gulf. World Appl. Sci. J. 2009, 7, 748–755.
  75. Cruickshank, R.; Duguid, J.P.; Marmion, B.P.; Swam, H.A. The Practice of Medical Microbiology, 12th ed.; Churchill Livingstone: London, UK, 1975; pp. 544–565.
  • Sample Availability: Samples of the compounds 2–4, 11, 18, 22, 24, 27, 33–36, 38, and 40 are available from the authors.
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