Microwave Synthesis, Characterization, and Antimicrobial Activity of Some Novel Isatin Derivatives

El-Faham, Ayman; Hozzein, Wael N.; Wadaan, Mohammad A. M.; Khattab, Sherine N.; Ghabbour, Hazem A.; Fun, Hoong-Kun; Siddiqui, Mohammed Rafiq

doi:https://doi.org/10.1155/2015/716987

Journal of Chemistry

On this page

Abstract Introduction Results and Discussion Conclusions Supplementary Materials References Copyright Related Articles

Research Article | Open Access

Volume 2015 | Article ID 716987 | https://doi.org/10.1155/2015/716987

Microwave Synthesis, Characterization, and Antimicrobial Activity of Some Novel Isatin Derivatives

Ayman El-Faham,^1,2Wael N. Hozzein,³Mohammad A. M. Wadaan,³Sherine N. Khattab,²Hazem A. Ghabbour,⁴Hoong-Kun Fun,^4,5and Mohammed Rafiq Siddiqui¹

Academic Editor: Artur M. S. Silva

Received27 Jun 2015

Accepted05 Aug 2015

Published27 Aug 2015

Abstract

Three series of isatin derivatives [3-hydrazino, 3-thiosemicarbazino, and 3-imino carboxylic acid derivatives] were synthesized employing microwave irradiation. The prepared compounds were characterized by FT-IR, NMR, elemental analysis, and X-ray crystallography for derivatives 5b. The synthesized compounds were screened for antimicrobial activity against selected bacteria and fungi. The results revealed that the N-alkyl isatin derivatives were biologically active with different spectrums activity. Most of the 3-hydrazino and 3-thiosemicarbazino isatin derivatives were biologically inactive and generally the active derivatives showed weak to moderate activity mainly against Gram-positive bacteria. The imino isatin carboxylic acid derivatives (2-[4-(1-benzyl-5-bromo-2-oxoindolin-3-ylideneamino) phenyl]acetic acid, 5d) showed promising activity against all tested Gram-positive bacteria and against fungal pathogens.

1. Introduction

The development of new therapeutic agents is one of the essential goals in medicinal chemistry. In the recent years there has been a spectacular increase in the use of microwave heating within the pharmaceutical industry, because it facilitates the synthesis of new chemical entities with reduced reaction time [1]. Microwave irradiation is considered as an alternate source of heating and it has been proven recently that microwave heating improves the efficiency of the reaction and reduces the reaction time [2–7].

It was reported that the biological properties of isatin derivatives include different effects on the brain and offer protection against some types of infections [8–11]. Isatin is considered as important class of bioactive compounds exhibiting caspase inhibitor [12, 13], antibacterial, and antiproliferative activity [14]. Schiff bases of isatin analogous have anti smallpox [15] and GAL3 receptor antagonist capabilities [16]. Isatin derivatives reported to show antiviral [17], anti-inflammatory, analgesic [18], and anticonvulsant activities [19]. Isatin--thiosemicarbazone derivatives were found to demonstrate a range of chemotherapeutic activities [20–26]. The literature survey revealed that thiocarbohydrazide is an irreversible inhibitor of catalase, which inhibits the degradation of serotonin and also possesses anticonvulsant activity [27].

We report here the synthesis of three different series of isatin derivatives (hydrazone, thiosemicarbazone, and imino carboxylic acid derivatives) employing microwave irradiation. The prepared compounds were subjected to in vitro screening for their antimicrobial activities against representatives of pathogenic Gram-positive and Gram-negative bacteria and fungi.

2. Experimental Section

2.1. Chemistry

2.1.1. Materials

The solvents used were of HPLC reagent grade. Melting points were determined with a Mel-Temp apparatus and are uncorrected. Fourier transform infrared spectroscopy (FTIR) spectra were recorded on a Nicolet 560 spectrometer. Nuclear magnetic resonance spectra (¹H NMR and ¹³C NMR) were recorded on a JEOL 400 MHz spectrometer with chemical shift values reported in ppm ( units) relative to an internal standard. The microwave irradiation was conducted using a multimode reactor (Synthos 3000, Anton Paar GmbH, and 1400 W maximum magnetron). Reactions were performed in teflon vessels (capacity 10 mL). The target temperature was set and fixed during the irradiation. Settings and readings for power (W) and pressure were taken from the instrument. Elemental analyses were performed on a Perkin-Elmer 2400 elemental analyzers, and the values found were within ±0.3% of the theoretical values. Follow-up of the reactions and checks of the purity of the compounds were done by thin layer chromatography (TLC) on silica gel-protected aluminum sheets (GF254, Merck), and the spots were detected by exposure to UV-lamp at 254 nm for a few seconds. The compounds were named using Chem. Draw Ultra version 11, Cambridge Soft Corporation. The X-ray diffraction measurements of compound 5b were performed using Bruker APEX-II D8 Venture diffractometer.

2.1.2. Synthesis of Isatin Derivatives (1b–1f)

A mixture of isatin (5 mmol) and potassium carbonate (8 mmol) in DMF (10 mL) was stirred for 10 minutes at room temperature. Alkyl halides (6 mmol; benzyl bromide for preparation of 1c, 1d, and 1e; CH₃I for preparation of 1b; and 1,3-dibromoethane for preparation of 1f) were added dropwise to the reaction mixture and then the reaction was microwave irradiated using a multimode reactor (Synthos 3000, Anton Paar GmbH, Graz, Austria) (1,400 W maximum magnetron). The vessels were heated for 5 minutes at 80°C and held at the same temperature for a further 5 minutes (400 W). Cooling was accomplished by a fan (5 minutes). The final product was dried and recrystallized from ethanol. All the spectral data for the products obtained were in good agreement with the reported data.

(1) 1-Methylisatin (1b). The product was obtained as orange red crystals in 90% yield; mp 126-127°C [Lit. [28] mp 126-127°C; in 73% yield]. ¹H NMR (CDCl₃): = 3.36 (s, 3H, CH₃), 6.78 (d, J = 7.8 Hz, 1H, Ar), 7.12 (t, J = 7.4 Hz, 1H, Ar), 7.47 (t, J = 8.1 Hz, 1H, Ar), 7.61 (d, J = 8.1 Hz, 1H, Ar) ppm.

(2) 1-Benzylisatin (1c). The product was obtained as orange red crystals in 88% yield; mp 134–136°C. [Lit. [28] mp 134–136°C; 88% yield]. ¹H NMR (CDCl₃): = 4.93 (s, 2H, C₆H₅CH₂), 6.77 (d, J = 7.7 Hz, 1H, Ar), 7.08 (t, J = 7.4 Hz, 1H, Ar), 7.33 (s, 5H, Ar), 7.47 (t, J = 8.1, 1H), 7.61 (d, J = 8.1, 1H, Ar) ppm.

(3) 1-Benzyl-5-bromoisatin (1d; Supporting Information Page 2 in Supplementary Material Available Online at http://dx.doi.org/10.1155/2015/716987). The product was obtained as red orange crystals in 93% yield; mp 149–151°C [Lit. [29] mp 152-153°C, in 95% yield]. ¹H NMR (CDCl₃): = 4.91 (s, 2H, C₆H₅CH₂), 6.69 (d, J = 8.1 Hz, 1H, Ar), 7.24–7.32 (m, 5H, Ar), 7.45 (dd, 1H, J = 8.1 Hz, 2.2 Hz, Ar), 7.56 (d, 1H, J = 2.2 Hz, Ar) ppm.

(4) 1-Benzyl-5-chloroisatin (1e; Supporting Information Page 3). The product was obtained as orange crystals in 86% yield; mp 136°C (Lit. [29] mp 134°C, in 65% yield]. ¹H NMR (CDCl₃): = 4.94 (s, 2H, C₆H₅CH₂), 6.69 (d, J = 8.1 Hz, 1H, Ar), 7.28–7.34 (m, 5H, Ar), 7.45 (dd, 1H, J = 8.1 Hz, 2.3 Hz, Ar), 7.56 (d, 1H, J = 2.3 Hz, Ar) ppm.

(5) 1-(2-Bromoethyl)isatin (1f; Supporting Information Page 4). The product was obtained as red orange crystals in 89% yield; mp 130-131°C (Lit. [29] mp 131-132°C; in 80% yield). ¹H NMR (CDCl₃): = 3.60 (t, 2H, J = 6.6 Hz, CH₂), 4.16 (t, 2H, J = 6.6 Hz, CH₂), 7.00 (d, 1H, J = 8.1 Hz, Ar), 7.16 (t, 1H, J = 7.3 Hz, Ar), 7.60–7.63 (m, 2H, Ar) ppm.

2.1.3. Microwave Method for the Synthesis of (3a–3n)

A multimode reactor (Synthos 3000 Anton Paar, GmbH, 1400 W maximum magnetron) was used. The initial step was conducted with 4-Teflon vessels rotor (MF 100) that allows the reactions to process under the same conditions. Isatin derivatives 1a–1f and hydrazine hydrate 2a, thiosemicarbazide 2b, or 4-aminobenzamide 2c were mixed together in a small portion of ethanol and then subjected to microwave irradiation (400 watt). The vessels were heated for 5 min at 100°C and held at the same temperature for another 5 min. cooling was accomplished by a fan (5 min). The final product was washed with cold ethanol and then dried under vacuum to afford the products 3a–3n in high yield and purity.

(1) 3-Hydrazinoisatin (3a; Supporting Information Pages 5 and 6). The product was obtained as yellow crystals from ethanol, in 91% yield; mp 240–242°C (Lit. [30] 242°C). ¹H NMR (d₆-DMSO): = 6.86 (d, J = 8.1 Hz, 1H, Ar), 6.98 (t, J = 7.3 Hz, 1H, Ar), 7.14 (t, J = 7.3 Hz, 1H, Ar), 7.35 (d, J = 8.1 Hz, 1H, Ar), 9.55 (d, J = 14.0 Hz, 1H, NH), 10.54 (d, J = 14.0 Hz, 1H, NH), 10.75 (brs, 1H, NH) ppm. ¹³C NMR (d₆-DMSO): = 110.5, 118.01, 121.9, 122.8, 126.8, 127.6, 139.2, 163.3 ppm.

(2) 1-Methyl-3-hydrazinoisatin (3b; Supporting Information Pages 7 and 8). The product was obtained as yellow crystals from ethanol in 93% yield, mp 95–97°C. ¹H NMR (d₆-DMSO): = 3.66 (s, 3H, CH₃), 6.96–7.05 (m, 2H, Ar), 7.23 (t, J = 8.0 Hz, 1H, Ar), 7.41 (d, J = 7.3 Hz, 1H, Ar), 9.64 (d, J = 8.8 Hz, 1H, NH), 10.58 (d, J = 10.3 Hz, 1H, NH) ppm. ¹³C NMR (d₆-DMSO): = 25.7, 109.8, 117.7, 121.9, 122.4, 125.9, 127.6, 140.5, 161.3 ppm. C₉H₉N₃O (175.19): Calcd. C, 61.70; H, 5.18; N, 23.99; found: C, 61.87; H, 5.33; N, 24.19.

(3) 1-Benzyl-3-hydrazinoisatin (3c; Supporting Information Pages 9 and 10). The product was obtained as yellow crystals from ethanol in 93% yield; mp 114-115°C (Lit. [30] 114-115°C). ¹H NMR (d₆-DMSO): = 4.97 (s, 2H, CH₂), 6.96 (d, J = 8.0 Hz, 1H, Ar), 7.02 (t, J = 7.3 Hz, 1H, Ar), 7.17 (t, J = 8.0 Hz, 1H, Ar), 7.32 (s, 5H, Ar), 7.43 (d, J = 7.3 Hz, 1H, Ar), 9.77 (d, J = 14.0 Hz, 1H, NH), 10.58 (d, J = 14.0 Hz, 1H, NH) ppm. ¹³C NMR (d₆-DMSO): = 42.8, 109.8, 117.9, 122.13, 122.6, 125.6, 127.5, 127.8, 127.9, 129.2, 137.3, 139.5, 161.3 ppm.

(4) 1-Benzyl-5-bromo-3-hydrazinoisatin (3d; Supporting Information Pages 11 and 12). The product was obtained as yellow crystals from ethanol in 95% yield; mp 134–136°C. ¹H NMR (d₆-DMSO): = 4.97 (s, 2H, CH₂), 6.96 (d, 1H, J = 8.1 Hz, Ar), 7.19 (d, 1H, J = 8.1 Hz, Ar), 7.24–7.33 (m, 5H, Ar), 8.15 (s, 1H, Ar), 9.98 (brs, 1H, 1NH), 10.35 (brs, 1H, NH) ppm. ¹³C NMR (d₆-DMSO): = 40.3, 111.3, 117.4, 123.9, 126.7, 127.0, 127.8, 128.0, 129.2, 136.9, 138.0, 161.1 ppm. C₁₅H₁₂BrN₃O (329.02): Calcd. C, 54.56; H, 3.66; N, 12.73; found: C, 54.78; H, 3.54; N, 12.98.

(5) 1-Benzyl-5-chloro-3-hydrazinoisatin (3e). The product was obtained as yellow crystals from ethanol in 93% yield; mp 105–107°C. ¹H NMR (d₆-DMSO-): = 4.97 (s, 2H, CH₂), 6.96 (d, J = 8.1 Hz, 1H, Ar), 7.19 (d, J = 8.1 Hz, 1H), 7.24–7.33 (m, 5H, C₆H₅), 8.15 (s, 1H, Ar), 9.98 (brs, 1H, 1NH), 10.35 (brs, 1H, NH) ppm; ¹³C NMR (d₆-DMSO): = 40.2, 111.3, 117.4, 123.9, 127.0, 127.8, 128.0, 129.2, 137.0, 138.0, 161.1 ppm. C₁₅H₁₂ClN₃O (285.73): Calcd. C, 63.05; H, 4.23; N, 14.71; found: C, 63.31; H, 4.44; N, 14.98.

(6) Isatin-3-thiosemicarbazone (3f). The product was obtained as yellow crystals in 89% yield; mp 246°C (dec) (Lit. [29] 245-246°C). ¹H NMR (d₆-DMSO): = 6.85–7.00 (m, 2H, Ar), 7.10 (brs, 2H, Ar), 7.50 (brs, 2H, NH₂), 8.05 (s, 1H, NH), 12.65 (brs, 1H, NH) ppm.

(7) 1-Benzyl isatin-3-thiosemicarbazone (3g; Supporting Information Pages 13 and 14). The product was obtained as yellow solid from ethanol in 93% yield; mp 246-247°C. ¹H NMR (d₆-DMSO): = 4.98 (s, 2H, CH₂), 7.02 (d, J = 8.1 Hz, 1H, Ar), 7.19 (t, J = 8.1 Hz, 1H, Ar), 7.24–7.39 (m, 5H, Ar), 7.72 (d, J = 8.1 Hz, 1H, Ar), 8.77 (s, 1H, 1NH), 9.12 (s, 1H, NH), 12.42 (s, 1H, 1NH) ppm. ¹³CNMR (d₆-DMSO): = 43.1, 110.9, 120.1, 121.4, 123.6, 128.0, 128.2, 129.3, 131.5, 131.6, 136.3, 143.2, 161.4, 179.3 ppm. C₁₆H₁₄N₄OS (310.37): Calcd. C, 61.92; H, 4.55; N, 18.05; found: C, 62.13; H, 4.68; N, 18.31.

(8) 1-Benzyl-5-bromoisatin-3-thiosemicarbazone (3h; Supporting Information Pages 15 and 16). The product was obtained as yellow solid from ethanol in 91% yield; mp 246-247°C. ¹H NMR (d₆-DMSO): = 4.98 (s, 2H, CH₂), 6.98 (d, J = 8.0 Hz, 1H, Ar), 7.24–7.38 (m, 5H, Ar), 7.52 (d, J = 8.0 Hz, 1H), 7.96 (s, 1H, Ar), 8.90 (s, 1H, 1NH), 9.18 (s, 1H, NH), 12.23 (s, 1H, 1NH) ppm. ¹³C NMR (d₆-DMSO): = 43.2, 112.9, 115.5, 122.4, 123.9, 122.4, 123.9, 128.0, 128.2, 129.3, 130.1, 133.5, 136.0, 142.1, 161.0, 179.3 ppm. C₁₆H₁₃BrN₄OS (389.27): Calcd. C, 49.37; H, 3.37; N, 14.39; found: C, 49.53; H, 3.61; N, 14.66.

(9) 1-Benzyl-5-chloroisatin-3-thiosemicarbazone (3i; Supporting Information Pages 17 and 18). The product was obtained as yellow solid from ethanol in 91% yield; mp 242°C (dec). ¹H NMR (d₆-DMSO): δ = 4.98 (s, 2H, CH₂), 7.02 (d, J = 8.1 Hz, 1H, Ar), 7.20–7.50 (m, 5H, Ar), 7.82 (s, 1H, Ar), 8.89 (s, 1H, 1NH), 9.19 (s, 1H, NH), 12.25 (s, 1H, 1NH) ppm. ¹³C NMR (d₆-DMSO) = 43.2, 112.4, 121.1, 122.0, 127.9, 128.0, 128.2, 129.3, 130.3, 130.8, 136.0, 141.7, 161.1, 179.3 ppm. C₁₆H₁₃ClN₄OS (344.82): Calcd. C, 55.73; H, 3.80; N, 16.25; found: C, 56.00; H, 3.89; N, 16.01.

(10) 1-(2-Bromoethyl)isatin-3-thiosemicarbazone (3j; Supporting Information Pages 19 and 20). The product was obtained as yellow crystals from ethanol in 88% yield; mp 246°C (dec). ¹H NMR (d₆-DMSO): = 3.77 (t, 2H, J = 6.4 Hz, CH₂), 4.21 (t, 2H, J = 6.4 Hz, CH₂), 7.18 (t, 1H, J = 7.3 Hz, Ar), 7.28 (d, 1H, J = 8.0 Hz, Ar), 7.43 (t, 1H, J = 7.3 Hz, Ar), 7.21 (d, 1H, J = 7.3 Hz, Ar), 8.77 (s, 1H, NH), 9.12 (s, 1H, NH), 12.36 (s, 1H, NH) ppm. ¹³C NMR (d₆-DMSO) = 30.1, 40.5, 110.9, 119.8, 121.3, 131.3, 131.8, 142.9, 161.3, 179.2 ppm. C₁₁H₁₁BrN₄OS (327.20): Calcd. C, 40.38; H, 3.39; N, 17.12; found: C, 40.66; H, 3.54; N, 17.40.

(11) 4-Amino--(2-oxoindolin-3-ylidene)benzohydrazide (3k; Supporting Information Pages 21 and 22). The product was obtained as a yellowish brown solid in 82% yield; mp > 250°C. ¹H NMR (d₆-DMSO): = 6.00 (brs, 2H, NH₂), 6.64 (d, 2H, J = 8.8 Hz, Ar), 6.92 (d, 1H, J = 8.8 Hz, Ar), 7.11 (t, 1H, J = 8.1 Hz, Ar), 7.38 (t, 1H, J = 8.1 Hz, Ar), 7.76 (d, 2H, J = 8.1 Hz, Ar), 7.88 (d, 1H, J = 7.3 Hz, Ar), 10.80 (s, 1H, NH), 11.33 (s, 1H, NH) ppm. ¹³C NMR (d₆-DMSO): = 111.1, 113.3, 116.5, 118.9, 122.4, 127.0, 131.0, 132.8, 142.2, 144.1, 153.7, 165.5 ppm. C₁₅H₁₂N₄O₂ (280.28): Calcd. C, 64.28; H, 4.32; N, 19.99; found: C, 64.13; H, 4.19; N, 20.14.

(12) 4-Amino--(1-methyl-2-oxoindolin-3-ylidene)benzohydrazide (3l; Supporting Information Pages 23 and 24). The product was obtained as a yellow powder from ethyl acetate in 88% yield; mp > 250°C. ¹H NMR (d₆-DMSO): = 3.26 (s, 3H, CH₃), 6.01 (brs, 2H, NH₂), 6.64 (d, 2H, J = 8.4 Hz, Ar), 7.12 (d, 1H, J = 8.0 Hz, Ar), 7.16 (d, 1H, J = 7.6 Hz, Ar), 7.38 (t, 1H, J = 7.6 Hz, Ar), 7.74 (d, 2H, J = 8.8 Hz, Ar), 7.88 (d, 1H, J = 7.6 Hz Ar), 11.34 (s, 1H, NH) ppm. ¹³C NMR (d₆-DMSO): = 26.0, 109.2, 112.7, 113.1, 115.2, 118.3, 122.2, 126.1, 131.1, 132.2, 139.1, 144.5, 153.7, 163.6 ppm. C₁₆H₁₄N₄O₂ (294.31): Calcd. C, 65.30; H, 4.79; N, 19.04; found: C, 65.45; H, 4.85; N, 19.31.

(13) 4-Amino--(1-benzyl-2-oxoindolin-3-ylidene)benzohydrazide (3m; Supporting Information Pages 25 and 26). The product was obtained as yellowish brown solid in 87% yield; mp 198–200°C. ¹H NMR (d₆-DMSO): = 4.99 (s, 2H, CH₂), 6.04 (brs, 2H, NH₂), 6.67 (d, 2H, J = 8.0 Hz, Ar), 7.00 (d, 1H, J = 7.3 Hz, Ar), 7.14 (t, 1H, J = 7.3 Hz, Ar), 7.27–7.41 (m, 5H, Ar), 7.65 (d, 1H, J = 8.8 Hz, Ar), 7.76 (d, 2H, J = 8.0 Hz, Ar), 7.95 (d, 1H, J = 7.3 Hz, Ar), 11.39 (s, 1H, NH) ppm. ¹³C NMR (d₆-DMSO) δ = 43.9, 110.3, 113.3, 113.7, 116.0, 123.0, 126.9, 127.7, 128.0, 128.1, 128.2, 129.3, 130.0, 131.2, 132.6, 142.3, 144.0, 153.8, 164.3 ppm. C₂₂H₁₈N₄O₂ (370.41): Calcd. C, 71.34; H, 4.90; N, 15.13; found: C, 71.60; H, 5.01; N, 15.41.

(14) 4-Amino--(1-benzyl-3-bromo-2-oxoindolin-3-ylidene)benzohydrazide (3n; Supporting Information Pages 27 and 28). The product was obtained as brown solid in 83% yield; mp 260–263°C. ¹H NMR (d₆-DMSO): = 4.99 (s, 2H, CH₂), 6.04 (brs, 2H, NH₂), 6.67 (d, 2H, J = 8.04 Hz, Ar), 7.00 (d, 1H, J = 7.3 Hz, Ar), 7.14 (t, 1H, J = 7.3 Hz, Ar), 7.27–7.41 (m, 5H, Ar), 7.65 (d, 1H, J = 8.8 Hz, Ar), 7.76 (d, 2H, J = 8.1 Hz, Ar), 7.95 (d, 1H, J = 7.3 Hz, Ar), 11.39 (s, 1H, NH), 11.33 (s, 1H, NH) ppm. ¹³C NMR (d₆-DMSO): = 43.9, 110.3, 113.3, 113.7, 116.0, 123.0, 126.9, 127.7, 128.0, 128.1, 128.2, 129.3, 130.0, 131.2, 132.6, 142.3, 144.0, 153.8, 164.3 ppm. C₂₂H₁₈N₄O₂ (449.30): Calcd. C, 71.34; H, 4.90; N, 15.13; found: C, 71.60; H, 5.01; N, 15.41.

2.1.4. Microwave Method for the Synthesis of (5a–5d)

Isatin derivatives 1a–1d and 4-aminophenyl acetic acid 4 were mixed in small amount of ethanol as a solvent in the presence of glacial acetic acid (2-3 drops). The individual vessels were heated for 5 min at 100°C and held at the same temperature for another 5 min; cooling was accomplished by a fan (5 min). The final product was washed with cold methanol and then dried under vacuum to afford products 5a–5d in a pure state, as observed from their spectral data.

(1) 2-[4-(2-Oxoindolin-3-ylideneamino)phenyl]acetic Acid (5a; Supporting Information Pages 29 and 30). The product was obtained as brown crystals from ethyl acetate in 88% yield; mp: 210–212°C. ¹H NMR (d₆-DMSO): = 3.63 (s, 2H, CH₂), 6.43 (d, 1H, J = 7.3 Hz, Ar), 6.72 (t, 1H, J = 7.3 Hz, Ar), 6.90 (d, 1H, J = 8.1 Hz, Ar), 6.98 (d, 2H, J = 8.1 Hz, Ar), 7.32–7.40 (m, 3H, Ar), 10.88 (s, 1H, NH) ppm. ¹³C NMR (d₆-DMSO): δ = 40.2, 112.1, 116.3, 117.9, 119.9, 122.2, 125.8, 129.8, 131.1, 132.3, 135.0, 147.5, 149.5, 155.4, 164.0, 173.3 ppm. C₁₆H₁₂N₂O₃ (280.28): Calcd. C, 68.56; H, 4.32; N, 9.99; found: C, 68.78; H, 4.49; N, 10.08.

(2) 2-[4-(1-Methyl-2-oxoindolin-3-ylideneamino)phenyl]acetic Acid (5b; Supporting Information Pages 31 and 32). The product was obtained as brown crystals from ethyl acetate in 88% yield; mp 198–200°C. ¹H NMR (d₆-DMSO): = 3.19 (s, 3H, CH₃), 3.62 (s, 2H, CH₂), 6.45 (d, 1H, J = 8.0 Hz, Ar), 6.78 (t, 1H, J = 8.0 Hz, Ar), 6.92 (d, 2H, J = 8.0 Hz, Ar), 7.08 (d, 1H, J = 8.0 Hz, Ar), 7.34 (d, 2H, J = 8.0 Hz, Ar), 7.43 (t, 1H, J = 8.0 Hz, Ar) ppm. ¹³C NMR (d₆-DMSO) = 26.1, 40.0, 110.2, 115.1, 117.3, 119.4, 122.2, 124.9, 130.6, 131.9, 134.4, 147.9, 148.8, 154.1, 162.2, 172.8 ppm. C₁₇H₁₄N₂O₃(294.30): Calcd. C, 69.38; H, 4.79; N, 9.52; found: C, 69.54; H, 4.86; N, 9.78.

(3) 2-[4-(1-Benzyl-2-oxoindolin-3-ylideneamino)phenyl]acetic Acid (5c; Supporting Information Pages 33 and 34). The product was obtained as reddish brown crystals from ethyl acetate in 86% yield; mp 180–182°C. ¹H NMR (d₆-DMSO): = 3.64 (s, 2H, CH₂), 4.99 (s, 2H, CH₂), 6.50 (d, 1H, J = 7.3 Hz, Ar), 6.78 (t, 1H, J = 8.0 Hz, Ar), 6.96–7.07 (m, 3H, Ar), 7.29–7.46 (m, 8H, Ar) ppm. ¹³C NMR (d₆-DMSO): = 40.3, 43.4, 111.2, 114.3, 115.9, 117.9, 122.9, 125.7, 127.9, 128.1, 129.2, 130.3, 131.2, 132.5, 134.8, 136.4, 147.4, 149.3, 154.4, 162.9, 173.3 ppm. C₂₃H₁₈N₂O₃ (370.40): Calcd. C, 74.58; H, 4.90; N, 7.56; found: C, 74.74; H, 5.05; N, 7.42.

(4) 2-[4-(1-Benzyl-5-bromo-2-oxoindolin-3-ylideneamino)phenyl]acetic Acid (5d). The product was obtained as reddish brown crystals from ethylacetate in 89% yield; mp 179-180°C. ¹H NMR (d₆-DMSO): = 3.65 (s, 2H, CH₂), 4.99 (s, 2H, CH₂), 6.64 (d, 1H, J = 7.3 Hz, Ar), 6.90–7.11 (m, 3H, Ar), 7.29–7.48 (m, 8H, Ar) ppm. ¹³C NMR (d₆-DMSO): = 40.3, 43.4, 114.3, 117.7, 117.9, 127.9, 128.0, 128.1, 129.2, 129.3, 131.2, 136.1, 136.8, 146.5, 148.9, 154.6, 162.9, 173.2 ppm. C₂₃H₁₇BrN₂O₃ (449.30): Calcd. C, 61.48; H, 3.81; N, 6.23; found: C, 61.73; H, 3.99; N, 6.51.

2.2. Biology

2.2.1. Antimicrobial Activities of the Prepared Compounds

The antimicrobial activities of the prepared compounds were tested against representative pathogenic Gram-positive, Gram-negative, and filamentous fungus test strains by using the paper disc diffusion method [31]. In this method, three agar media, nutrient agar [32] for bacteria, Sabouraud agar [33] for the yeast strain, and Czapek Dox agar for the fungus strain [34], were prepared and sterilized by autoclaving at 120°C and 1.5 atm for 20 min. The agar plates were poured, left to cool down and after solidification they were inoculated with the bacterial and fungal strains by streaking. The synthesized compounds were dissolved in dimethylsulfoxide (DMSO) at a final concentration of 10 mg/mL and 5 µL of each compound was loaded on a sterile filter paper disc (5 mm diameter; 50 µg per disc). The filter paper disc was then transferred aseptically into the inoculated agar plates along with commercially available tetracycline and erythromycin discs as positive controls and DMSO as the negative control for comparison. The plates were then incubated at 37°C for 24 h for bacteria and at 30°C for 48–72 h for fungi. After incubation, diameters of the inhibition zones around the paper discs were measured in mm as an indication of the antimicrobial activities of the compounds.

3. Results and Discussion

3.1. Chemistry

N-alkyl isatin derivatives 1b–1f were prepared from isatin derivatives according to the reported method employing microwave irradiation [30]. The spectral data of 1b–1f were in good agreement with the reported data [28–30, 35–38]. Isatin derivatives 3a–3n were prepared by condensation of 1a–1f either with hydrazine hydrate 2a, thiosemicarbazide 2b, or 4-aminobenzhydrazide 2c following the reported procedure using microwave irradiation (Scheme 1) [30]. The products 3a–3n were obtained in excellent yields (85–95%; Section 2) and their spectral data were in good agreement with their proposed structures.

The IR spectra of 3a–3e showed absorption bands in the regions 3398 to 3217 cm⁻¹ corresponding to (NH), a band at 1688 cm⁻¹ corresponding to the C=O group, and a peak around 1584 cm⁻¹ that reveals the formation of the C=N bond. The NMR spectra of all the products 3a–3e are in a good agreement with their structures [28–30].

The IR spectra of the series 3f–3j showed absorption bands in the region 3453 cm⁻¹ related to the asymmetric (N–H) vibration of the terminal NH₂ group. The other bands at 3336 and 3256 cm⁻¹ may be due to the symmetric (N–H) vibrations of the amide and amino groups. The peaks at 1680 and 1588 cm⁻¹ correspond to C=O and C=N group, respectively. The NMR of all the products 3f–3j are in good agreement with their proposed structures (Section 2).

The IR spectra of 3k–3n reveal absorption bands in the regions 3406, 3325, and 3225 cm⁻¹ which can be assigned to the asymmetric (N–H) vibration of the terminal NH₂ group and the amide groups. The peaks at 1712, 1642, and 1592 cm⁻¹ correspond to two C=O and C=N groups, respectively. NMR spectra of all the obtained products 3k–3n are in good agreement with their proposed structures (Section 2).

The imino isatin carboxylic acid derivatives 5a–5d were prepared by condensation of isatin derivatives 1a–1d with 4-aminophenylacetic acid 4 using microwave irradiation and ethanol as a solvent (Scheme 2).

The IR spectra of the compounds derived from 4-aminophenylacetic acid 5a–5d reveal broad absorption bands in the region 3432-2632 cm⁻¹ which can be assigned to the hydroxyl group of the carboxylic group. The peaks at 1729, 1645, and 1602 cm⁻¹ are related to the (C=O) of the carboxylic, carbonyl, and imino groups, respectively. The ¹H NMR and ¹³C-NMR spectra of the products 5a–5d are also in good agreement with their structures. The ¹H NMR spectrum of compound 5b as a prototype displayed two singlet peaks at δ 3.19 and δ 3.62 corresponding to CH₃ and the benzylic protons (C₆H₅CH₂), respectively. Six peaks corresponding to the 8 aromatic protons were observed at δ = 6.45, 6.78, 6.92, 7.08, 7.34, and 7.43 ppm. The ¹³C-NMR spectrum confirmed its structure, where two signal were observed at δ = 26.1 and 40.0 ppm assigned to the methyl and benzylic protons, respectively. The observed peak at δ = 154.1, 162.2, and 172.8 ppm is related to the C=N, C=O, and the carbonyl of the carboxylic groups, respectively.

Single crystal X-ray crystallography is without doubt a decisive analytical tool which confirms the configuration of the imine double bond in the target compounds 5a–5d. Fortunately, we have succeeded in getting single crystals of compound 5b that was suitable for X-ray crystallography. The solved structure is a representative example for the synthesized target compounds 5a–5d. The assigned (E)-configuration of compound 5b was established via its single crystal X-ray structure (Figure 1).

Molecular formula of 5b is as follows: C₁₇H₁₄N₂O₃, Molecular weight: 294.30, Monoclinic, P21/c, a = 10.3860 (8) Å, b = 8.5519 (7) Å, = 16.0149 (11) Å, = 96.604 (2)°, V = 1413.01 (19) Å³, calc = 1.383 Mg m⁻³, orange plate with 0.69 × 0.35 × 0.08 mm. A total of 28282 reflections were measured, of which 3241 were independent. int = 0.163, dataset (; ; ) = −13, 13; −11, 11; −20, 20. Refinement of against all reflections, led to , , and = 1.07.

Crystallographic data have been deposited with the Cambridge Crystallographic Data Center (supplementary publication number CCDC-1004251). Copies of the data may be obtained free of charge from the Director, CCDC, 12 Union Road, Cambridge, UK.

3.2. Biology

The antimicrobial activities of the prepared compounds against a panel of pathogenic tested organisms are collected in Table 1. The results revealed that all the N-alkyl isatin derivatives 1b–1f were biologically active with different spectrum activity. The isatin itself 1a exhibited average antibacterial activities against the Gram-negative test organisms only, but the activity decreased with the presence of the methyl group in 1b. On the other hand, the introduction of a benzyl group in 1c decreased the activity against Gram-negative bacteria and confers activity against Gram-positive bacteria, namely, Bacillus subtilis. However, the presence of ethyl bromide group in 1f expands the spectrum activity to be active against Bacillus subtilis and two Gram-negative tested bacteria and against the fungal pathogens in study.

Differently, most of the 3-hydrazino and 3-thiosemicarbazino isatin derivatives 3a–3n were biologically inactive and generally the active derivatives showed weak to moderate activity against at least two of the tested Gram-positive test bacteria. Interestingly, the activity was associated with the presence of the bromine or chlorine in the isatin moieties 3d, 3e, and 3n; however, their effect cannot be confirmed in the activity of 3h and 3i because 3f was originally active.

Among the imino isatin carboxylic acid derivatives, 5d which has bromine at position 5 showed good to high antimicrobial activity compared to the positive control (erythromycin and tetracycline) against the Gram-positive and fungal test organisms. The presence of the methyl group in 5b was associated with the activity against the Gram-negative bacteria and, similarly as in the N-alkyl isatin derivatives, compound 5c, having a benzyl group, showed antimicrobial activity against Bacillus subtilis and Candida albicans. These results obtained are similar to the reported results by Singh et al. and Patel et al. [28, 39].

4. Conclusions

Three series of isatin derivatives (3-hydrazino, 3-thiosemicarbazino, and 3-imino carboxylic acid derivatives) were synthesized by employing microwave irradiation in good yields and purities. The prepared compounds were evaluated for their antimicrobial activity against Gram-positive and Gram-negative bacteria and fungal pathogenic organisms. In general, the N-alkyl isatin derivatives were biologically active, while some of the 3-hydrazino and 3-thiosemicarbazino isatin derivatives were active against at least two of the three used Gram-positive test organisms. It was found that the activity was associated with the presence of the bromine or chlorine in the isatin moiety of the prepared compounds. The highest activity was achieved with one of the imino isatin carboxylic acid derivatives that showed good to high activity towards the three Gram-positive bacteria and the two fungal tested microorganisms.

Conflict of Interests

The authors declare no conflict of interests.

Acknowledgment

The authors thank the King Abdul-Aziz City for Science and Technology (KACST) in the Kingdom of Saudi Arabia for funding this work through project “SGP-34-55.”

Supplementary Materials

¹H and ¹³CNMR spectra.

Supplementary Material

References

R. W. Wagner, “Microwave-assisted synthesis in the pharmaceutical industry—a current perspective and future prospects,” Drug Discovery World, vol. 7, no. 3, pp. 59–66, 2006.
View at: Google Scholar
J. L. Krstenansky and L. Cotterill, “Recent advances in microwave-assisted organic syntheses,” Current Opinion in Drug Discovery and Development, vol. 3, no. 4, pp. 454–461, 2000.
View at: Google Scholar
G. Kiran, T. Maneshwar, Y. Rajeshwar, and M. Sarangapani, “Microwave-assisted synthesis, characterization, antimicrobial and antioxidant activity of some new Isatin derivatives,” Journal of Chemistry, vol. 2013, Article ID 192039, 7 pages, 2013.
View at: Publisher Site | Google Scholar
M. de Rosa and A. Soriente, “A combination of water and microwave irradiation promotes the catalyst-free addition of pyrroles and indoles to nitroalkenes,” Tetrahedron, vol. 66, no. 16, pp. 2981–2986, 2010.
View at: Publisher Site | Google Scholar
G.-R. Qu, J. Wu, Y.-Y. Wu, F. Zhang, and H.-M. Guo, “Synthesis of novel 6-[N,N-bis(2-hydroxyethyl)amino]purine nucleosides under microwave irradiation in neat water,” Green Chemistry, vol. 11, no. 6, pp. 760–762, 2009.
View at: Publisher Site | Google Scholar
D. Dallinger and C. O. Kappe, “Microwave-assisted synthesis in water as solvent,” Chemical Reviews, vol. 107, no. 6, pp. 2563–2591, 2007.
View at: Publisher Site | Google Scholar
H. M. Meshram, N. N. Rao, N. S. Kumar, and L. C. Rao, “Microwave assisted catalyst free synthesis of 3-hydroxy-2-oxidoles by aldol condensation of acetophenones with isatins,” Der Pharma Chemica, vol. 4, no. 3, pp. 1355–1360, 2012.
View at: Google Scholar
F. M. D. Joaquim, J. G. Simon, and C. P. Angelo, “The chemistry of isatin: a review from 1975–1999,” Journal of the Brazilian Chemical Society, vol. 12, pp. 273–324, 2001.
View at: Google Scholar
V. Glover, S. K. Bhattacharya, and M. Sandler, “Isatin a new biological factor,” Indian Journal of Experimental Biology, vol. 29, pp. 1–5, 1991.
View at: Google Scholar
S. Ghosal, S. K. Bhattacharya, A. V. Muruganandam, and K. S. Satyan, “Iron ion-catalyzed oxidation of tryptophan—a simulated biosynthetic route to isatin,” Biogenic Amines, vol. 13, no. 2, pp. 91–100, 1997.
View at: Google Scholar
S. N. Pandeya, S. Smitha, M. Jyoti, and S. K. Sridhar, “Biological activities of isatin and its derivatives,” Acta Pharmaceutica, vol. 55, no. 1, pp. 27–46, 2005.
View at: Google Scholar
W. Chu, J. Zhang, C. Zeng et al., “N-benzylisatin sulfonamide analogues as potent caspase-3 inhibitors: synthesis, in vitro activity, and molecular modeling studies,” Journal of Medicinal Chemistry, vol. 48, no. 24, pp. 7637–7647, 2005.
View at: Publisher Site | Google Scholar
W. Chu, J. Rothfuss, Y. Chu, D. Zhou, and R. H. Mach, “Synthesis and in vitro evaluation of sulfonamide isatin Michael acceptors as small molecule inhibitors of caspase-6,” Journal of Medicinal Chemistry, vol. 52, no. 8, pp. 2188–2191, 2009.
View at: Publisher Site | Google Scholar
Z. H. Chohan, H. Pervez, A. Rauf, K. M. Khan, and C. T. Supuran, “Isatin-derived antibacterial and antifungal compounds and their transition metal complexes,” Journal of Enzyme Inhibition and Medicinal Chemistry, vol. 19, no. 5, pp. 417–423, 2004.
View at: Publisher Site | Google Scholar
M. C. Pirrung, S. V. Pansare, K. Das Sarma, K. A. Keith, and E. R. Kern, “Combinatorial optimization of isatin-β-thiosemicarbazones as anti-poxvirus agents,” Journal of Medicinal Chemistry, vol. 48, no. 8, pp. 3045–3050, 2005.
View at: Publisher Site | Google Scholar
M. J. Konkel, B. Lagu, L. W. Boteju et al., “3-Arylimino-2-indolones are potent and selective galanin GAL₃ receptor antagonists,” Journal of Medicinal Chemistry, vol. 49, no. 13, pp. 3757–3758, 2006.
View at: Publisher Site | Google Scholar
A. Jarrahpour, D. Khalili, E. De Clercq, C. Salmi, and J. M. Brunel, “Synthesis, antibacterial, antifungal and antiviral activity evaluation of some new bis-Schiff bases of isatin and their derivatives,” Molecules, vol. 12, no. 8, pp. 1720–1730, 2007.
View at: Publisher Site | Google Scholar
O. A. Sharaf, “Some pharmacological activities of new substituted pyrolloindoles, indolothiazepine and azole derivatives,” Bulletin of Faculty of Pharmacy, vol. 35, pp. 79–82, 1997.
View at: Google Scholar
M. Verma, S. N. Pandeya, K. N. Singh, and J. P. Stables, “Anticonvulsant activity of Schiff bases of isatin derivatives,” Acta Pharmaceutica, vol. 54, no. 1, pp. 49–56, 2004.
View at: Google Scholar
Y. Teitz, D. Ronen, A. Vansover, T. Stematsky, and J. L. Riggs, “Inhibition of human immunodeficiency virus by N-methylisatin-β4′:4′-diethylthiosemicarbazone and N-allylisatin-β-4′:4′-diallythiosemicarbazone,” Antiviral Research, vol. 24, no. 4, pp. 305–314, 1994.
View at: Publisher Site | Google Scholar
M. D. Hall, N. K. Salam, J. L. Hellawell et al., “Synthesis, activity, and pharmacophore development for isatin-β-thiosemicarbazones with selective activity toward multidrug-resistant cells,” Journal of Medicinal Chemistry, vol. 52, no. 10, pp. 3191–3204, 2009.
View at: Publisher Site | Google Scholar
H. Beraldo and D. Gambino, “The wide pharmacological versatility of semicarbazones, thiosemicarba-zones and their metal complexes,” Mini-Reviews in Medicinal Chemistry, vol. 4, no. 1, pp. 31–39, 2004.
View at: Publisher Site | Google Scholar
I.-J. Kang, L.-W. Wang, T.-A. Hsu et al., “Isatin-β-thiosemicarbazones as potent herpes simplex virus inhibitors,” Bioorganic and Medicinal Chemistry Letters, vol. 21, no. 7, pp. 1948–1952, 2011.
View at: Publisher Site | Google Scholar
S. Bajroliya, G. S. Kalwania, S. Choudhary, and S. Chomal, “Synthesis, characterization and antimicrobial activities of 1,2,4-triazole /isatin Schiff bases and their Mn(II), Co(II) Complexes,” Oriental Journal of Chemistry, vol. 30, no. 4, pp. 1601–1608, 2014.
View at: Publisher Site | Google Scholar
A. M. Naglah, H. M. Awad, M. A. Bhat, M. A. Al-Omar, and A. E. Amr, “Microwave-assisted synthesis and antimicrobial activity of some novel isatin Schiff bases linked to nicotinic acid via certain amino acid bridge,” Journal of Chemistry, vol. 2015, Article ID 364841, 8 pages, 2015.
View at: Publisher Site | Google Scholar
J. Azizian, M. K. Mohammadi, O. Firuzi, N. Razzaghi-Asl, and R. Miri, “Synthesis, biological activity and docking study of some new isatin Schiff base derivatives,” Medicinal Chemistry Research, vol. 21, no. 11, pp. 3730–3740, 2012.
View at: Publisher Site | Google Scholar
K. Gangarapu, S. Manda, A. Jallapally et al., “Synthesis of thiocarbohydrazide and carbohydrazide derivatives as possible biologically active agents,” Medicinal Chemistry Research, vol. 23, no. 2, pp. 1046–1056, 2014.
View at: Publisher Site | Google Scholar
U. K. Singh, S. N. Pandeya, A. Singh, B. K. Srivastava, and M. Pandey, “Synthesis and antimicrobial activity of Schiff's and N-Mannich bases of isatin and its derivatives with 4-amino-N-carbamimidoyl benzene sulfonamide,” International Journal of Pharmaceutical Sciences and Drug Research, vol. 2, no. 2, pp. 151–154, 2010.
View at: Google Scholar
M. O. Shibinskaya, S. A. Lyakhov, A. V. Mazepa et al., “Synthesis, cytotoxicity, antiviral activity and interferon inducing ability of 6-(2-aminoethyl)-6H-indolo[2,3-b]quinoxalines,” European Journal of Medicinal Chemistry, vol. 45, no. 3, pp. 1237–1243, 2010.
View at: Publisher Site | Google Scholar
A. El-Faham, A. A. Elzatahry, Z. A. Al-Othman, and E. A. Elsayed, “Facile method for the synthesis of silver nanoparticles using 3-hydrazino-isatin derivatives in aqueous methanol and their antibacterial activity,” International Journal of Nanomedicine, vol. 9, no. 1, pp. 1167–1174, 2014.
View at: Publisher Site | Google Scholar
E. J. De Beer and M. B. Sherwood, “The paper-disc agar-plate method for the assay of antibiotic substances,” Journal of Bacteriology, vol. 50, pp. 459–467, 1945.
View at: Google Scholar
S. T. Williams and T. Cross, “Actinomycetes,” in Methods in Microbiology, C. Booth, Ed., pp. 295–335, Academic Press, London, UK, 1971.
View at: Google Scholar
F. C. Odds, “Sabouraud('s) agar,” Journal of Medical and Veterinary Mycology, vol. 29, no. 6, pp. 355–359, 1991.
View at: Publisher Site | Google Scholar
R. M. Atlas, Handbook of Microbiological Media, CRC Press, Boca Raton, Fla, USA, 2nd edition, 1997.
G. Chen, H. J. Su, M. Zhang et al., “New bactericide derived from Isatin for treating oilfield reinjection water X-ray data collection and structure refinement,” Chemistry Central Journal, vol. 6, pp. 90–97, 2012.
View at: Google Scholar
G. Chen, Y. Wang, X. Hao, S. Mu, and Q. Sun, “Simple isatin derivatives as free radical scavengers: synthesis, biological evaluation and structure-activity relationship,” Chemistry Central Journal, vol. 5, no. 1, article 37, 2011.
View at: Publisher Site | Google Scholar
J. Azizian, H. Fallah-Bagher-Shaidaei, and H. Kefayati, “A facile one-pot method for the preparation of N-Alkyl isatins under microwave irradiation,” Synthetic Communications, vol. 33, no. 5, pp. 789–793, 2003.
View at: Publisher Site | Google Scholar
C. M. Clay, H. M. Abdallah, C. Jordan, K. Knisley, and D. M. Ketcha, “N-alkylation of isatins utilizing KF/alumina,” Arkivoc, vol. 2012, no. 6, pp. 317–325, 2012.
View at: Publisher Site | Google Scholar
A. Patel, S. Bari, G. Talele, J. Patel, and M. Sarangapani, “Synthesis and antimicrobial activity of some new isatin derivatives,” Iranian Journal Pharmaceutical Research, vol. 4, pp. 249–254, 2006.
View at: Google Scholar

Copyright

Copyright © 2015 Ayman El-Faham et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

PDF Download Citation

Download other formats

Order printed copies

Views

3613

Downloads

1730

Citations