Editor-in-Chief Hatice Kübra Elçioğlu Vice Editors Levent Kabasakal Esra Tatar Online ISSN 2630-6344 Publisher Marmara University Frequency Bimonthly (Six issues / year) Abbreviation J.Res.Pharm. Former Name Marmara Pharmaceutical Journal
Journal of Research in Pharmacy 2013 , Vol 17 , Num 2
Synthesis and chemotherapeutic activities of 5-chloro-1H-indole-2,3-dione 3-thiosemicarbazones
Görkem Ermut1, Nilgün Karalı1, İdil Çetin2, Mehmet Topçul3, Seher Birteksöz4
1Istanbul University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry, Istanbul, Turkey
2Istanbul University, Institute of Science, Department of Biology, Istanbul, Türkiye
3Istanbul University, Faculty of Science, Department of Biology, Istanbul, Turkey
4Istanbul University, Faculty of Pharmacy, Department of Pharmaceutical Microbiology, Istanbul, Turkey
DOI : 10.12991/201317383

Summary

Bir seri 5-kloro-1H-indol-2,3-dion 3-tiyosemikarbazon 3a-g, kemoterapötik etkilerini incelemek için sentezlenmiştir. 3a-g nin yapıları spektral bulgular (IR, 1H NMR, 13C NMR-APT, 13C NMR-DEPT, HSQC, HMBC) ve elementel analiz ile kanıtlanmıştır. Bileşiklerin insan serviks kanserinden türetilmiş HeLa hücrelerine antikanser etkileri hücre kinetik parametreleri kullanılarak test edilmiştir. Ön inceleme sonuçları alkil sübstitüe 3a, 3b ve 3d’ nin sentez evresine kıyasla mitoz evresinde daha etkili olduğunu göstermiştir. 3a-g CRFK, HeLa, HEL, MDCK ve Vero hücrelerinde bazı DNA ve RNA virüslerine karşı araştırılmıştır. Bileşiklerin hiçbiri DNA ve RNA virüslerine karşı etkili değildir. Tüm bileşiklerin seçilen suşlara karşı antimikrobial etkileri test edilmiştir. Metil sübstitüe 3a ve allil sübstitüe 3c’ nin S. aureus and C. albicans’a karşı etkili olduğu bulunmuştur.

Introduction

Biological properties of thiosemicarbazone derivatives have been studied since 1946, when the activity of thiacetazone against Mycobacterium tuberculosis was reported. Since then, this and other biological properties of thiosemicarbazone derivatives, such as anticancer and antiviral activity have been described. Triapine is a ribonucleotide reductase inhibitor with promising anticancer activity against hematologic malignancies in clinical trials[1]. Isatin-Β-thiosemicarbazone (IBT) and N-methylisatin-Β-thiosemicarbazone (MIBT, methisazone) prevent the production of small-pox viruses. Methisazone has also been used in the clinical treatment of smallpox[2]. N-methylisatin-Β,4’,4’-diethylthiosemicarbazone (M-IBDET) specifically inhibits formation of Moloney leukemia virus structural proteins[3] (Figure 1).


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FIGURE 1: Representative bioactive thiosemicarbazone derivatives

Isatin (1H-indole-2,3-dione) is a synthetically versatile molecule which has led to an array of derivatives displaying a broad spectrum of biological properties[4,5]. Investigation of the structure- activity relationships in 3-substituted 2-indolinone derivatives revealed that halogenation at the 5-position and 3-thiosemicarbazone formation were associated with increased activity against a range of human cancer cell lines, various bacteria and viruses[6-10]. Selective activity toward multidrug resistant (MDR) cells of several thiosemicarbazone derivatives were recently tested. Pharmacophore analysis of active compounds revealed that isatin-3-thiosemicarbazone moiety was essential for the MDR1-selective activity[10,11].

In the light of these findings, 5-chloro-1H-indole- 2,3-dione 3-thiosemicarbazone derivatives were synthesized in order to obtain more potent and less toxic compounds. The structures of the synthesized compounds were determined by analytical and spectral (IR, 1H NMR, 13C NMR-APT, 13C NMR-DEPT, HSQC- 2D, HMBC-2D) methods. The compounds were tested to investigate their the chemotherapeutic activities.

Methods

Chemistry
Chemicals and reagents used in the current study were of analytical grade. Melting points were estimated with a Buchi 540 melting point apparatus in open capillaries and are uncorrected. Elemental analyses were performed on a Thermo Finnigan Flash EA 1112 elemental analyzer. IR spectra were recorded on KBr discs, using a Perkin-Elmer Model 1600 FT-IR spectrometer. 1H NMR, 13C NMR-APT, 13C NMR-DEPT, HSQC-2D and HMBC-2D spectra were obtained on VarianUNITY INOVA (500 MHz) or Varian Mercury (300 MHz) spectrophotometers using DMSO-d6.

The synthesis of 4-substituted thiosemicarbazides (1a-g)
To a solution of hydrazine hydrate (5 mmol) in ethanol (10 mL), a suspension of an appropriate isothiocyanate (5 mmol) in ethanol (10 mL) was added dropwise with vigorous stirring and cooling in an ice bath. The mixture was allowed to stand overnight. The crystals formed were recrystallized from ethanol.

The synthesis of 5-chloro-1H-indole-2,3-dione 3-thiosemicarbazones (3a-g)
A solution of 4-substituted thiosemicarbazides 1a-g (3.5 mmol) in ethanol (10 mL) was added to a solution of 5-chloro-1H-indole- 2,3-dione 2 (3.5 mmol) in ethanol (20 mL). After addition of a drop of concentrated sulfuric acid, the mixture was refluxed on a water bath for 5 h. The product formed after cooling was filtered and washed with ethanol or recrystallized from ethanol.

5-Chloro-1H-indole-2,3-dione 3-(4-methylthiosemicarbazone) (3a) (12)
Yield: 95%, mp: 254oC, IR (KBr) cm-1: υ 3264 (NH), 1697 (C=O), 1142 (C=S); 1H NMR (DMSO-d6, 500 MHz) ppm: δ 3.07 (d, J= 4.40 Hz, 3H, CH3), 6.91 (d, J= 8.30 Hz, 1H, indole C7-H), 7.35 (dd, J= 8.29, 1.95 Hz, 1H, indole C6-H), 7.67 (d, J= 1.95 Hz, 1H, indole C4-H), 9.33 (q, J= 4.88 Hz, 1H, N4-H), 11.26 (s, 1H, indole NH), 12.39 (s, 1H, N2-H); HSQC-2D (DMSO- d6, 125 MHz) ppm: δ 31.98 (CH3), 113.23 (indole C7), 120.98 (indole C4), 122.62 (indole C3a), 127.19 (indole C5), 130.98 (indole C6), 130.99 (indole C7a), 141.55 (indole C3), 163.07 (indole C=O), 178.37 (C=S). Anal. Cald for C10H9ClN4OS (268.72): C, 44.70; H, 3.38; N, 20.85%. Found: C, 44.70; H, 3.85; N, 20.95%.

5-Chloro-1H-indole-2,3-dione 3-(4-ethylthiosemicarbazone) (3b)
Yield: 94%, mp: 267ºC, IR (KBr) cm-1: υ 3365, 3255 (NH), 1689 (C=O), 1153 (C=S). 1H NMR (DMSO-d6, 300 MHz) ppm: δ 1.18 (t, J= 7.19 Hz, 3H, ethyl C2-H), 3.62 (quin., J= 6.82 Hz, 2H, ethyl C1-H), 6.92 (d, J= 8.69 Hz, 1H, indole C7- H), 7.36 (dd, J= 8.25, 2.25 Hz, 1H, indole C6-H), 7.71 (d, J= 2.09 Hz, 1H, indole C4-H), 9.38 (t, J= 5.85 Hz, 1H, N4-H), 11.29 (s, 1H, indole NH), 12.36 (s, 1H, N2-H). Anal. Cald for C11H11ClN4OS (282.75): C, 46.73; H, 3.92; N, 19.82%. Found: C, 46.41; H, 3.79; N, 19.62%.

5-Chloro-1H-indole-2,3-dione 3-(4-allylthiosemicarbazone) (3c)
Yield: 96%, mp: 215ºC, IR (KBr) cm-1: υ 3356, 3253 (NH), 1693 (C=O), 1170 (C=S); 1H NMR (DMSO-d6, 500 MHz) ppm: δ 4.25 (t, J= 4.87 Hz, 2H, allyl C1-H), 5.14 (dd, J= 10.38, 1.52 Hz, 1H, allyl C3-H cis), 5.20 (dd, J= 17.22, 1.52 Hz, 1H, allyl C3- Htrans), 5.87-5.95 (m, 1H, allyl C2-H), 6.93 (d, J= 8.23 Hz, 1H, indole C7-H), 7.36 (dd, J= 8.39, 2.29 Hz, indole C6-H), 7.74 (d, J= 2.44 Hz, 1H, indole C4-H), 9.53 (t, J= 5.79 Hz, 1H, N4-H), 11.27 (s, 1H, indole NH), 12.42 (s, 1H, N2-H). Anal. Cald for C12H11ClN4OS.1/2H2O (303.77): C, 47.44; H, 3.97; N, 18.44%. Found: C, 47.58; H, 3.96; N, 18.33%.

5-Chloro-1H-indole-2,3-dione 3-(4-butylthiosemicarbazone) (3d)
Yield: 97%, mp: 224ºC, IR (KBr) cm-1: υ 3251 (NH), 1693 (C=O), 1169 (C=S). 1H-NMR (DMSO-d6, 300 MHz) ppm: δ 0.91 (t, J= 7.64 Hz, 3H, butyl C4-H), 1.30-1.38 (m, 2H, butyl C3-H), 1.60 (quin., J= 7.49 Hz, 2H, butyl C2-H), 3.59 (q, J= 7.49 Hz, 2H, butyl C1-H), 6.93 (d, J= 8.39 Hz, 1H, indole C7-H), 7.37 (dd, J= 8.25, 2.25 Hz, 1H, indole C6-H), 7.73 (d, J= 2.1 Hz, 1H, indole C4-H), 9.37 (t, J= 5.99 Hz, 1H, N4-H), 11.29 (s, 1H, indole NH), 12.36 (s, 1H, N2-H). Anal. Cald for C13H15ClN4OS (310.80): C, 50.24; H, 4.86; N, 18.03%. Found: C, 50.47; H, 4.98; N, 17.97%.

5-Chloro-1H-indole-2,3-dione 3-(4-benzylthiosemicarbazone) (3e)
Yield: 93%, mp: 245ºC, IR (KBr) cm-1: υ 3373, 3159 (NH), 1691 (C=O), 1161 (C=S). 1H-NMR (DMSO-d6, 500 MHz) ppm: δ 4.87 (d, J= 6.1 Hz, 2H, benzyl CH2), 6.92 (d, J= 8.24 Hz, 1H, indole C7-H), 7.25-7.37 (m, 6H, indole C6-H, C6H5), 7.71 (d, J= 2.13 Hz, 1H, indole C4-H), 9.90 (t, J= 6.25 Hz, 1H, N4-H), 11.29 (s, 1H, indole NH), 12.48 (s, 1H, N2-H). 13C NMR (APT, DMSO-d6, 75 MHz) ppm: δ 47.9 (benzyl CH2), 113.01 (indole C7), 120.95 (indole C4), 122.34 (indole C3a), 126.95 (indole C5), 127.53 (phenyl C3,5), 127.74 (phenyl C2,6), 128.78 (phenyl C4), 130.87 (indole C6), 131.29 (indole C7a), 138.71 (phenyl C1), 141.43 (indole C3), 162.4 (indole C=O), 178.15 (C=S). Anal. Cald for C16H13ClN4OS. H2O (362.84): C, 52.96; H, 4.16; N, 15.44%. Found: C, 53.13; 4.28; 15.58%.

5-Chloro-1H-indole-2,3-dione 3-[4-(4-fluorophenyl)thiosemicarbazone] (3f)[13]
Yield: 97%, mp: 236ºC, IR (KBr) cm-1: υ 3326, 3215 (NH), 1690 (C=O), 1171 (C=S). 1H-NMR (DMSO-d6, 300 MHz) ppm: δ 6.92 (d, J= 8.40 Hz, 1H, indole C7-H), 7.26 (t, J= 8.89 Hz, 2H, phenyl C3,5-H), 7.35 (d, J= 2.10 Hz, 1H, indole C6-H), 7.59 (dd, J= 8.99, 5.09 Hz, 2H, phenyl C2,6-H), 7.82 (d, J= 1.80 Hz, indole C4-H), 10.87 (s, 1H, N4-H), 11.34 (s, 1H, indole NH), 12.60 (s, 1H, N2- H). 13C NMR DEPT, HMBC-2D (DMSO-d6, 75 MHz) ppm: δ 113.01 (indole C7), 115.65 (d, J= 22.47 Hz, phenyl C3,5), 121.41 (indole C4), 122.22 (indole C3a), 127.03 (indole C5), 128.43 (d, J= 8.67 Hz, phenyl C2,6), 131.07 (indole C6), 131.58 (indole C7a), 135.08 (phenyl C1), 141.59 (indole C3), 158.91 (d, J= 242.99 Hz, phenyl C4), 162.89 (indole C=O), 177.09 (C=S). Anal. Cald for C15H10ClFN4OS (348.78): C, 51.65; H, 2.89; N, 16.06%. Found: C, 51.79; H, 2.99; N, 16.06%.

5-Chloro-1H-indole-2,3-dione 3-[4-(4-nitrophenyl)thiosemicarbazone] (3g)
Yield: 91%, mp: 258ºC, IR (KBr) cm-1: υ 3286, 3212 (NH), 1699 (C=O), 1179 (C=S). 1H-NMR (DMSO-d6, 300 MHz) ppm: δ 6.94 (d, J= 8.10 Hz, 1H, indole C7-H), 7.40 (dd, J= 8.40, 2.10 Hz, 1H, indole C6-H), 7.85 (d, J= 2.09 Hz, 1H, indole C4-H), 8.06 (d, J=9.29 Hz, 2H, phenyl C2,6-H), 8.29 (d, J= 9.79 Hz, 2H, phenyl C3,5- H), 11.10 (s, 1H, N4-H), 11.38 (s, 1H, indole NH), 12.79 (s, 1H, N2-H). 13C NMR (APT, DMSO-d6, 75 MHz) ppm: δ 113.15 (indole C7), 116.98 (indole C4), 121.61 (indole C3a), 122.0 (phenyl C2,6), 124.48 (phenyl C3,5), 127.07 (indole C5), 131,45 (indole C6), 132.49 (indole C7a), 141.85 (indole C3), 144.78 (phenyl C1), 146.99 (phenyl C4), 162.95 (indole C=O), 176.38 (C=S). Anal. Cald for C15H10ClN5O3S (375.78): C, 47.94; H, 2.68; N, 18.64%. Found: C, 47.43; H, 2.81; N, 18.35%.

CYTOTOXICITY
Cell Culture
The HeLa cell line used in this experiment was obtained from European Cell Culture Collection (CCL). Cells were cultured in Medium-199 (M-199, Sigma, USA) containing 10% fetal bovine serum (FBS, Gibco Lab), 100 μg/mL streptomicin (Streptomicin sulphate, I. E. Ulugay), 100 IU/ml penicilin (Pronapen, Pfizer), Amphotericine B (Sigma, USA) and 2 mM glutamine at 37ºC in humidified atmosphere of 5% CO2 in air. The pH of the medium was adjusted to 7.4 with NaHCO3.

Drug application
Drugs used in this study were dissolved immediately in DMSO before the preparation of the required concentrations. We used optimum doses of all drugs. Optimum doses were obtained by dilution of the stock solution. These doses were 1 mg/mL for 3a, 3e and 3g; 10 mg/mL for 3b and 3d. The drugs were tested by using these doses and HeLa cells were treated with these doses in the time periods of 24, 48 and 72 h.

Mitotic index analysis
Mitotic index was studied by the methods of Feulgen. Before the cells were treated with Feulgen, they were treated with 1 N HCl at room temperature for 1 min and then hydrolized with 1 N HCl for 10.5 min at 60°C. After slides were treated with Feulgen, they were rinsed for a few minutes in distilled water and stained with 10% Giemsa stain solution (pH 6.8) for 3 min and washed twice in phosphate buffer. After staining, the slides were rinsed in distilled water. And then the slides were air dried. At last mitotic index was calculated by counting metaphases, anaphases and telophases for each tested drug concentration and control. At least three thousand cells were examined from each slide for mitotic index (14).

3H-thymidine labelling index
At the end of drug administration, to investigate the parameter of labelling index, cells were treated with the medium containing 1 μCi/mL 3H-thymidine for 20 min.

Autoradiography
After labelling, the cells were fixed with Carnoys fixative [ethanol: glacial acetic acid (3: 1)] and remaining radioactive materials were washed twice with 2% perchloric acid at 4oC for 30 mins. After preparing slides, they were coated with K.2 gel emulsion (Ilford, England) prepared with distilled water at 40ºC to determine the thymidine labelling index. After 3 days of exposure at 4ºC, autoradiograms were washed with D- 19 b developer (Kodak) and fixed with Fixaj B (Kodak). The slides were evaluated after being stained with Giemsa for 3 minutes. On each slide, with 100x12.5 magnification in 100 areas, the labelled cells were counted. The same person evaluated all the slides by counting at least 3000 cells from each slides. These data are typical results from a minimum of three independent experiments[15].

Results

Chemistry
In this study, hydrazine hydrate was reacted with an appropriate isothiocyanate in ethanol to give the corresponding 4-substituted thiosemicarbazides 1a-g[16-18]. A series of 5-chloro-1H-indole-2,3-dione 3-thiosemicarbazones 3a-g were synthesized by reacting 5-chloro-1H-indole-2,3-dione 2 with 1a-g in ethanol containing a catalytic amount of sulphuric acid[7-9]. The structures of 3a-g were confirmed by analytical and spectral (IR, 1H NMR, 13C NMR-APT, 13C NMR-DEPT, HSQC- 2D, HMBC-2D) data (Scheme).


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SCHEME 1: Synthesis of compounds 3a-g. Reagents and conditions: i) EtOH, stirred, cooled ii) EtOH, H2SO4, reflux, 6 h.

IR spectra of 3a-g showed absorption bands in the 3373-3159 cm-1 region resulting from the NH stretchings of the lactam and thioamide functions. The lactam C=O and thioamide C=S stretchings were observed in the 1699-1689 and 1179-1142 cm-1 regions, respectively. The IR spectra provided evidence for the confirmation of the thiosemicarbazone structure. Amidic and ketonic C=O stretching bands of 5-chloro-1H-indole-2,3-dione 2 absorbing as two separate bands absorbed as a single amidic C=O stretching band and a ketonic C=O stretching band disappeared in the IR spectra of 3a-g[19,20]. 1H NMR spectra of 3a-g displayed the NH protons of the thiosemicarbazone moiety (δ 9.33-11.10 and 12.36-12.79 ppm) and the indole NH proton (δ 11.26-11.38 ppm) as three separate signals. HSQC spectra of 3a, 13C NMR-APT spectra of 3e and 3g, 13C NMR-DEPT and HMBC spectra of 3f supported the IR and 1H-NMR findings, and displayed signals at δ 141.43-141.85, 162.4-163.07 and 176.38-178.37 ppm which were attributed to the quarternary indole C3, indole C2 and C=S atoms[7-9].

Chemotherapeutic activity
The anticancer activities of 3b, 3d, 3e and 3g, along with previously reported 3a on HeLa cells derived from human cervix carcinoma were evaluated using cell kinetic parameters including mitotic index and labelling index. Mitotic index is the ratio of the number of cells undergoing mitosis (cell division) to the number of cells not undergoing mitosis in a population of cells. 3H thymidine labelling index explains the ratio of cells with DNA synthesis to all the cells in growing cell populati on. Four different doses (D1= 1 mg/ mL, D2= 5 mg/mL, D3= 10 mg/mL and D4= 50 mg/mL) were prepared for the compounds and the optimum dose was determined using mitotic index parameter for all of them. Optimum doses were determined as D1 for 3a, 3e and 3g, and D3 for 3b and 3d. Then, they were applied to HeLa cells for 0-72 h. For both parameters all the differences between control and experimental groups were statistically significant (p<0.01). Sunitinib was used as the standard in the tests of the mitotic index (Table 1 and Figure 2)[21]. The oxindole derived sunitinib was approved by the FDA in January 2006 for the treatment of gastroinestinal stromal cancers and renal cell carcinoma[22].

TABLE 1: Mitotic index (%) values of HeLa cells treated with 10μM of Sunitinib


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FIGURE 2: Mitotic index (%) values of HeLa cells treated with 10μM of Sunitinib

3a, 3b and 3d showed varying degrees of inhibition at optimum doses in the mitotic index tests as can be seen in Tables 2-4 and Figures 3-5. For ethyl substituted 3b, an inhibition percentage of 50.41% was obtained. The inhibition percentages of methyl substituted 3a and n-butyl substituted 3d were 38.19% and 40.55%, respectively. In benzyl substituted 3e and 4-nitrophenyl substituted 3g, the activity significantly decreased when compared with the alkyl substituted derivatives. The inhibition percentages of the tested compounds were lower than the values observed for sunitinib.

TABLE 2: Mitotic index (%) values of HeLa cells treated with D1 (1 μg/mL) dose of 3a

TABLE 3: Mitotic index (%) values of HeLa cells treated with D3 (10 μg/mL) dose of 3b

TABLE 4: Mitotic index (%) values of HeLa cells treated with D3 (10 μg/mL) dose of 3d


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FIGURE 3: Mitotic index (%) values of HeLa cells treated with D1 (1 μg/mL) dose of 3a


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FIGURE 4: Mitotic index (%) values of HeLa cells treated with D3 (10 μg/mL) dose of 3b


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FIGURE 5: Mitotic index (%) values of HeLa cells treated with D3 (10 μg/mL) dose of 3d

In all experimental groups, labelling index values of 0-72 h were determined. The preliminary screening results indicated that all tested compounds were less effective in synthesis stage when compared to the mitosis phase (Tables 5-7 and Figures 6-8). n-Butyl substituted 3d displayed the highest efficacy among the compounds and the inhibition percentage was 39.05% for 3d.

TABLE 5: Labelling index (%) values of HeLa cells treated with D1 (1 μg/mL) dose of 3a

TABLE 6: Labelling index (%) values of HeLa cells treated with D3 (10 μg/ mL) dose of 3b

TABLE 7: Labelling index (%) values of HeLa cells treated with D3 (10 μg/ mL) dose of 3d


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FIGURE 6: Labelling index (%) values of HeLa cells treated with D1 (1 μg/mL) of 3a


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FIGURE 7: Labelling index (%) values of HeLa cells treated with D3 (10 μg/mL) dose of 3b


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FIGURE 8: Labelling index (%) values of HeLa cells treated with D3 (10 μg/mL) dose of 3d

3b-e and 3g, along with previously reported 3a and 3f were evaluated against feline corona virus (FIPV), feline herpes virus (FHV) in Crandell-Rees feline kidney (CRFK), parainfluenza- 3 virus, rheovirus-1, sindbis virus, coxsackie virus B4, punto toro virus in VERO, herpes simplex virus-1 (KOS)(HSV- 1), herpes simplex virus-2 (G)(HSV-2), vaccinia virus, vesicular stomatitis virus (VSV), herpes simplex virus-1 TK KOS ACV in human embroyonic lung (HEL) and vesicular stomatitis virus, coxsackie virus B4 and respiratory syncytial virus (RSV) in Henrietta Lacks (HeLa) cell cultures[23]. None of the test compounds was active against any of the RNA or DNA viruses, including influenza virus at 100 µM.

3b-e and 3g, along with previously reported 3a and 3f were evaluated against Staphylococcus aureus ATCC 6538, Staphylococcus epidermidis ATCC 12228, Escherichia coli ATCC 25922, Klebsiella pneumoniae ATCC 4352, Pseudomonas aeruginosa ATCC 27853, Proteus mirabilis ATCC 14153 ve Candida albicans ATCC 10231. The minimum inhibitory concentrations (MIC) of 3a and 3c were determined using a microdilution assay. Ciprofloxazin and clotrimazole were used as the standards in the tests (Table 12)[24,25]. The antimicrobial activity results show that methyl substituted 3a and allyl substituted 3c have considerable antimicrobial effect on S. aureus and C. albicans.

TABLE 8: The MIC values of 3a and 3c

Conclusion

A series of 5-chloro-1H-indole-2,3-dione 3-thiosemicarbazones were synthesized to investigate their chemotherapeutic activities. The compounds were evaluated for cytotoxic activities on HeLa cells derived from human cervix carcinoma. Replacement of the aralkyl or aryl at the R with alkyl have been found to yield more active compounds in mitosis phase, whereas none of the compounds was selective in the synthesis phase. The substitution of the methyl or allyl group at the R caused an increase in inhibitory activity against S. aureus and C. albicans. In conclusion, structural modification may lead to new derivatives with high selectivity for a range of human cancer cell lines, various bacteria and viruses.

ACKNOWLEDGEMENTS
We thank Prof. Lieve Naesens from the Rega Institute for Medical Research, Katholieke Universiteit, Leuven, Belgium for evaluation of anviral activity. This work was supported by Istanbul University Scientific Research Projects. Project Number: BYP-15286

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