Synthesis, Characterization, Antimicrobial and Antitumor Activity of 4-(6-aryl-5H-[1,2,4]triazolo[3,4-b][1,3]thiazin-3-yl) Aniline

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1 RESEARCH ARTICLE Synthesis, Characterization, Antimicrobial and Antitumor Activity of 4-(6-aryl-5H-[1,2,4]triazolo[3,4-b][1,3]thiazin-3-yl) Aniline Vikas G. Rajurkar 1*, Vinayak K. Deshmukh 2, Madhuri V. Gosavi 2, Rajesh B. Patil 3 1 Dr. Vedprakash Patil Pharmacy College, Paithan Road, Gevrai, Aurangabad , Aurangabad, Maharashtra, India 2 Department of Pharmaceutical Chemistry, MES`s College of Pharmacy (Affiliated To Savitribai Phule Pune University), Sonai , Ahmednagar, Maharashtra, India 3 Smt. Kashibai Navale College of Pharmacy, Kondhawa (Bk), Pune , Maharashtra, India. ABSTRACT:Literature reveals [1,2,4]triazolo[3,4-b][1,3]thiazin nucleus containing compounds are significant antimicrobial and anticancer drug candidates. Novel [1,2,4]triazolo[3,4-b][1,3]thiazin candidates (Va-k) were synthesized, characterized and tested for their antimicrobial and antitumor activities. Compounds Va, Ve, Vg, Vh, Vi and Vk showed potent and broad spectrum antimicrobial activity as compared to standard drug. Compound Va, Vb, Vc, Vf, and Vj were tested, found active against four out of sixty human tumor cell lines. Keywords : Triazolo; Thiazin; Antimicrobial; Anti cancer 1. Introduction Brisk increase in pathogenic fungi and bacteria that are multi resistant to chemotherapy has become paramount menace in treatment of contagious diseases. Heterocyclic compounds with 1,2,4-triazolo ring system substituted with nitrogen and sulfur manifest in numerous biological activities and as a result, the literature is abundant with reports on the cytotoxic potency of 1,2,4-triazolo-[3,4-b]- 1,3,4-thiadiazoles against various cancer cell lines [1-3],other activities such as antibacterial [4-6], anti oxidant [7-9], analgesic [10], nematicidal [11], anthelmintic [12], anti-inflammatory [13-14], anti viral [15]. The cytotoxic potency of 1,2,4-triazolo[3,4-b]-1,3,4-thiadiazoles against various cancer cell lines has been extensively reported [16-20]. Due to ambidentate nucleophilic centers, the 4-amino-5- aryl-4h-1,2,4-triazole-3-thiol are excellent starting materials for the synthesis of N-bridged heterocycles such as 7H-[1,2,4]triazolo[3,4-b][1,3,4]thiadiazine derivatives. The substituted 1,2,4-triazolo[3,4-b]-1,3,4-thiadiazine derivatives shows antimicrobial activity due to N C S liaison, in the skeleton of triazolo-thiadiazine and also acquire anticancer activity [21-22]. * Rajurkar V. G. Professor, Department of Pharmaceutical Chemistry Dr. Vedprakash Pharmacy College, Aurangabad vikas_rajurkar_1973@yahoo.co.in Phone ; Experimental 2.1 Materials All the reagents and solvents used in the present study were of analytical grade and purchased from Loba chemie (India).The progression of the reactions were examined by Thin Layer Chromatography using Merck silica gel precoated plates, with appropriate mobile phase, visualizing technique by iodine and Ultra Violet chamber and compounds were refined by recrystallization technique. All the melting points were recorded on a Veego apparatus (Mumbai, India) and are uncorrected. All the compounds were characterized by their FT-IR, 1 H- NMR, GC Mass spectroscopy. FT-IR spectra were recorded in KBr on Bruker FT-IR instrument (Germany), 1 H-NMR spectra were recorded on Bruker Avance 1 H- NMR spectrometer (Germany), 400 MHz in DMS-d6, by Varian instrument using TMS (tetramethylsilane) as internal standard and chemical shift values are given in ppm downfield to TMS and GC Mass were recorded on GCMS-QP-5050 Shimadzu (Japan) using Atmospheric Pressure Photoionization method. The reference standards Norfloxacin, and Ketoconazole were obtained as gift samples from Macleod s Pharma. Ltd., Daman, India. 2.2 Synthesis of ethyl-4-amino benzoate (I) In a 50 ml round bottom flask, p-amino benzoic acid (0.01 mol, 1.37 g) was dissolved into 20 ml dry ethanol, while stirring; 0.5 ml of conc. H2S4 was added dropwise to obtain precipitate and further reaction mixture was refluxed on water bath for 01 h, cooled to room Published by DYPIPSR, Pimpri, Pune ( MH ) INDIA 30

2 temperature. Cooled reaction solution was neutralized with 10 % Na2C3 until the ph was approximately 8 and extracted twice with 10 ml portions of methylene chloride. Methylene chloride layers were washed twice with 8 ml water; dried over anhydrous sodium sulfate and organic solvent removed using rotary evaporator, dried, recrystalized using a solvent pair, ethyl alcohol and water (1:1) and then air dried. M.P: oc, Yield % (Scheme I) [23]. 2.3 Synthesis of 4-aminobenzoic acid hydrazide (II) In a 50 ml round bottom flask, ethyl 4-aminobenzoate I (0.01 mol, 1.65g) and hydrazine hydrate 99 % (0.01mol, 3 ml) were dissolved into 20 ml ethanol; further refluxed on water bath for 01 h and allowed to stand overnight. The precipitated obtained was filtered, washed with water, dried and recrystalized using methanol. M.P: oc, Mobile phase- Ethyl acetate: Diethyl Ether: Water (3.1: 1.2: 5.7), Yield % (Scheme I) [24]. 2.4 Synthesis of potassium-4-amino phenyldithiocarbazate (III) In a 250mL round bottom flask, 4-aminobenzoic acid hydrazide II (0.075mol, g) was dissolved in a solution of KH (0.075 mol, 4.2g) in 100 ml of absolute alcohol and CS2 (0.01 mol) and further mixture was agitated overnight; diluted with 200 ml of dry ether. The solid obtained was filtered and washed with dry ether, Yield % (Scheme I) [25]. 2.5 Synthesis of 4-amino-5-(4-aminophenyl)-4H-1,2,4 - triazole-3-thiol (IV) A mixture of potassium-pyridine-dithiocarbazate III (0.01mol, 2.65 g) and hydrazine hydrate (0.015 mol, 4.5mL) was refluxed on oil bath at 180 o C for 02 h with occasional shaking and then reaction mixture was poured into the cold water; acidified with HCl. The precipitate obtained was filtered, dried and recrystalized using ethanol, M.P C, Yield % (Scheme I) [25]. 2.6 Synthesis of 4-[6-aryl-7H-[1,2,4]triazolo[3,4-b] [1,3,4]-thiadiazine-3-yl] aniline (V) A suspension of 4-amino-5-(4-aminophenyl)-4H-1,2,4- triazole-3-thiol IV (0.01 mol, 2.07 g) and α-bromo ketones (0.13 mol) in 20 ml of absolute ethanol was refluxed on water bath for 03 h, then anhydrous sodium acetate (0.011 mol, 0.9 g) was added and further refluxed for an additional 01 h, cooled; poured into ice-cold water, filtered, dried and recrystalized from ethanol (Scheme I) [26]. Mobile phase Ethanol (5mL): Ethyl acetate (2mL): drop of acetic acid [3-(4-aminophenyl)-7H-[1,2,4]triazolo[3,4-b] [1,3,4]thiadiazin-6-yl]phenol Va Yield: 66.60%, M.P.: oc. FT-IR ν max (KBr, cm -1): 770 (o-sub. phenyl),840 (p-sub. phenyl), 1278 (C=N str, ring), 1454,1504,1604 (C=C str, ring), 2868 (C-NH2, ring), 3080 (C-H str, aromatic), 3757(-H str, ring), 1H- NMR (DMS, 400 MHz): δ 4.3 (s,2h,methylene), 6.60 (s, 2H, amino), (m, 8H, aromatic), (s,1h, hydroxyl) MS:[M]+ at m/z 323. Analytical Calculations for Formula; C16H13N5S, Predicted: C, 59.43; H, 4.05; N, 21.66, Found: C, 59.45; H, 4.03; N, [3-(4-aminophenyl)-7H-[1,2,4]triazolo[3,4-b] [1,3,4]thiadiazin-6-yl]phenol Vb Yield: %, M.P.: oc. FT-IR ν max (KBr, cm -1): 826 (p-disub.ring), 1288 (C=N str, ring), 1455, 1503, 1604 (C=C str, ring), 2866 (C-NH2 str, ring), 3080 (C-H str, aromatic), 3742 (-H str, ring), 1H-NMR (DMS, H 2 N H p-amino benzoic acid EtH, H 2 S 4 Na 2 C 3 Sod.carbonate H 2 N Ethyl-4-amino benzoate I C 2 H 5 NH 2 NH 2.H 2 EtH,Reflux H 2 N NHNH 2 4-amino benzoic acid hydrazide II CS 2, KH Stirr Ether, EtH NH 2 N N H NH NH S 2 N HCl, NH 2 NH 2.H 2 EtH, Reflux N S N NH HS Reflux SK 2 H 2 N N alfa-bromo ketone N N 4-amino-5-(4-aminophenyl)- potassium-pyridine-dithiocarbazate III R 4H-1,2,4-triazole-3-thiol IV 4-[6-aryl-7H-[1,2,4]-triazolo[3,4-b] [1,3,4]-thiadiazine-3-yl]aniline Va-k Scheme I: Synthesis of 4-[6-aryl-7H-[1,2,4]triazolo[3,4-b][1,3,4]-thiadiazine-3-yl]aniline (Va-k) Published by DYPIPSR, Pimpri, Pune ( MH ) INDIA 31

3 400 MHz): δ 4.33 (s,2h, methylene), 6.43 (s, 2H, amino), (m, 8H, aromatic), (s,1h, hydroxyl) MS: [M]+ at m/z 323. Analytical Calculations for Formula; C16H13N5S, Predicted: C, 59.43; H, 4.05; N, 21.66, Found: C, 59.40; H, 4.06; N, [6-(4-chlorophenyl)-7H-[1,2,4]triazolo[3,4-b] [1,3,4]thiadiazin-3-yl]aniline Vc Yield: 67.25%, M.P.: oC. FT-IR ν max (KBr, cm- 1): 817(C-Cl str, p-disub.ring), 1297,1454 (C=C str, ring), 1606 (C=N str, ring), 3090 (C-H str, aromatic), 3723 (C- NH2 str, ring), 1H-NMR (DMS, 400 MHz): δ 4.35 (s,2h,methylene), 5.28 (s, 2H, amino), (m, 8H, aromatic) MS:[M]+ at m/z 341. Analytical Calculations for Formula; C16H12ClN5S, Predicted: C, 56.22; H, 3.54; N, 20.49, Found: C, 56.19; H, 3.51; N, [6-(4-bromophenyl)-7H-[1,2,4]triazolo[3,4-b] [1,3,4]thiadiazin-3-yl]aniline Vd Yield: %, M.P.: oc. FT-IR ν max (KBr, cm -1): 829 (C-Br str, p-disub.ring), 1287 (C=N str, ring), 1457, 1506,1588,1606 (C=C str, ring), 2988 (C-H str, aromatic), 2951 (C-NH2 str, ring) 1H-NMR (DMS, 400 MHz): δ 4.37 (s,2h,methylene), 4.91 (s, 2H, amino), (m, 8H, aromatic), MS:[M]+ at m/z 386. Analytical Calculations for Formula; C16H12BrN5S, Predicted: C, 49.75; H, 3.13; N, 18.13, Found: C, 49.77; H, 3.10; N, [6-(4-methylphenyl)-7H-[1,2,4]triazolo[3,4-b] [1,3,4]thiadiazin-3-yl]aniline Ve Yield: %, M.P.: oc. FT-IR ν max (KBr, cm -1): 826 (C-CH3 str, p-disub.ring), 1260 (C=N str, ring), 1453 (C-CH3, ring), 1604 (C=C str, ring), 3050 (C-H str, aromatic), 3831 (C-NH2, ring) 1H-NMR (DMS, 400 MHz): δ 2.27 (s,3h, methyl), 4.14 (s,2h,methylene), 6.03 (s, 2H, amino), (m, 8H, aromatic) MS:[M]+ at m/z 321. Analytical Calculations for Formula; C17H15N5S, Predicted: C, 63.53; H, 4.70; N, 21.79, Found: C, 63.49; H, 7.72; N, [6-(4-methoxyphenyl)-7H-[1,2,4]triazolo[3,4-b] [1,3,4]thiadiazin-3-yl]aniline Vf Yield: %, M.P.: oc. FT-IR ν max (KBr, cm -1): 827 (p-sub. ring), 1175(-CH3 str, ring), 1260 (C=N str, ring), 1456, 1486, 1566 (C=C str, ring), 2929 (C-NH2 str, ring), 3130 (C-H str, aromatic) 1H-NMR (DMS, 400 MHz): δ 3.92 (s,3h, methoxy), 4.42 (s, 2H, methylene), 6.60 (s, 2H, amino), (m, 8H, aromatic) MS:[M] + at m/z 337. Analytical Calculations for Formula; C17H15N5S, Predicted: C, 60.52; H, 4.48; N, 20.76, Found: C, 60.50; H, 4.46; N, [6-(4-nitrophenyl)-7H-[1,2,4]triazolo[3,4-b] [1,3,4]thiadiazin-3-yl]aniline Vg Yield: %, M.P.: oc. FT-IR ν max (KBr, cm -1): 840 (p-sub. ring), 1278 (C=N str, ring), 1344 ( C- N2, ring), 1454, 1504, 1580, 1604 (C=C str, ring), 2923 (C-H str, aromatic), 3075 (C-NH2, ring) 1H-NMR (DMS, 400 MHz): δ 4.40 (s,2h, methylene), 5.50 (s, 2H, amino), (m, 8H, aromatic) MS:[M]+ at m/z 352. Analytical Calculations for Formula; C16H12N62S, Predicted: C, 54.54; H, 3.43; N, 23.85, Found: C, 54.56; H, 3.40; N, ,4'-(7H-[1,2,4]triazolo[3,4-b][1,3,4]thiadiazine- 3,6-diyl)dianiline Vh Yield: %, M.P.: oc. FT-IR ν max (KBr, cm -1): 850 (p-sub. ring), 1257 (C=N str, ring), 1455, 1604 (C=C str, ring), 3010, 3080 (C-NH2 str, ring), 3105 (C-H str, aromatic) 1H-NMR (DMS, 400 MHz): δ 4.30 (s,2h, methylene), 5.20 (s, 2H, amino), (m, 8H, aromatic) MS:[M]+ at m/z 322. Analytical Calculations for Formula; C16H14N6S, Predicted: C, 59.61; H, 4.38; N, 26.07, Found: C, 59.59; H, 4.40; N, [6-(3-nitrophenyl)-7H-[1,2,4]triazolo[3,4-b] [1,3,4]thiadiazin-3-yl]aniline Vi Yield: %, M.P.: oc. FT-IR ν max (KBr, cm -1): 729 (o-sub. ring), 820 (p-sub. ring), 1342 (C=N str, ring), 1349 (o-n2 str, ring), 1402, 1503, 1528, 1609 (C=C str, ring), 3105 (C-H str, aromatic), 3045 (C-NH2 str, ring) 1H-NMR (DMS, 400 MHz): δ 4.40 (s,2h, methylene), 5.30 (s, 2H, amino), (m, 8H, aromatic), MS:[M]+ at m/z 352. Analytical Calculations for Formula; C16H12N62S, Predicted: C, 54.54; H, 3.43; N, 23.85, Found: C, 54.55; H, 3.41; N, [3-(4-aminophenyl)-7H-[1,2,4]triazolo[3,4 -b][1,3,4]thiadiazin-6-yl]-2h-chromen-2- one Vj Yield: %, M.P.: o C. FT-IR ν max (KBr, cm -1 ): 831 (p-sub. ring),1183 (C--C str, ring), 1303 (C=N str, ring), 1455, 1602 (C=C str, ring), 1715 (C= str, chromen), 2841(C-H str, aromatic), 1 H-NMR (DMS, 400 MHz): δ 4.35 (s,2h, methylene), 5.80 (s, 2H, amino), (m, 9H, aromatic) MS:[M]+ at m/z 375. Analytical Calculations for Formula; C 19 H 13 N 5 2 S, Predicted: C, 60.79; H, 3.49; N, 18.66, Found: C, 60.80; H, 3.47; N, [6-(biphenyl-4-yl)-7H-[1,2,4]triazolo[3,4- b][1,3,4]thiadiazin-3-yl]aniline Vk Yield: %, M.P.: o C. FT-IR ν max (KBr, cm -1 ): 809 (p-sub. ring), 1240 (C=N str, ring), 1489, 1603 (C=C str, ring), 2930 (C-H str, aromatic) 1 H-NMR (DMS, 400 MHz): δ 5.56 (s,2h, methylene), 6.57 (s, 2H, amino), (m, 13H, aromatic) MS:[M]+ at m/ z 383. Analytical Calculations for Formula; C 22 H 17 N 5 S, Predicted: C, 68.91; H, 4.47; N, 18.26, Found: C, 68.94; Published by DYPIPSR, Pimpri, Pune ( MH ) INDIA 32

4 H, 4.45; N, Biological Evaluation Antimicrobial evaluation All the synthesized compounds (Va-k) were screened for in vitro antimicrobial activity against two gram positive species S.aureus (S.aureus, NCIM 2079), B.subtillis (B.subtillis, NCIM 2711) and two gram negative strains E.coli (E.coli, NCIM 2685), P. aeruginosa (P. aeruginosa, NCIM 27853) and two fungal species A.nigar (A.nigar, NCIM 596), C.albicans (C.albicans, NCIM 18804) for antifungal evaluation, using the broth dilution method. Minimum inhibitory concentration (MIC) was determined and compared with standard reference Norfloxacin for antibacterial and Ketoconazole for antifungal evaluation (Table 1) [27, 28] In vitro anticancer evaluation The human tumor cell lines were developed in Roswell Park Memorial Institute medium 1640, inoculated into 96 well micro titer plates; incubated at 37 C (5 % C 2, 95 % air and 100 % RH) for 24 h. Then two plates of each cell line were fixed in situ with TCA, and cell population for each cell line (Tz) was recorded. Test compounds were dissolved in DMS to acquire the desired 10-5 M test concentration and stored frozen. At the time of drug addition, an aliquot of frozen concentrate was diluted to twice the desired 10-5 M test concentration with medium containing 50 µg/ml gentamicin. Aliquots of 100 µl of each compound were loaded to the appropriate micro titer wells containing 100 µl of medium and further plates were incubated for 48 h. Cells were fixed in situ by the addition of 50 µl of cold 50 % (w/v) of 10% TCA and incubated for 60 minutes at 4 C. The supernatant was discarded, and the plates were washed five times with water and air dried. 100 µl Sulforhodamine B (SRB) solutions at 0.4 % (w/v) in 1 % acetic acid was added to each well, and incubated for 10 min at RT, plates were then washed with 1 % acetic acid and air dried. Bound stain was subsequently solubilized with 10 M trizma base, and the absorbance was read on an automated plate reader at a wavelength of 515 nm [time zero, (Tz), control growth, (C), and test growth in the presence of test compound (Ti)]. Percentage growth inhibition was calculated using formula 1 a, b, c, d and e [29,30]. [(Ti-Tz)/(C-Tz)] x 100 for concentrations for which Ti>/ =Tz... -1a [(Ti-Tz)/Tz] x 100 for concentrations Ti<Tz... -1b Growth inhibition of 50 % (GI50) = [(Ti-Tz)/(C- 100 = c Total growth inhibition (TGI) = Ti = Tz -1d for which Tz)] x The LC50 (50% reduction in the measured protein) indicating a net loss of cells upon treatment = [(Ti- Tz)/ Tz] x 100 = e 3. Results and Discussion 3.1. Chemistry The scheme for synthesis of target compounds Va-k outlined in Scheme 1. Ethyl-4-amino benzoate (I) was synthesized from p-amino benzoic acid by esterification and later on exposed to condensation with 99% NH 2 NH 2.H 2 in the presence of EtH to acquire p-amino benzoic acid hydrazide (II). Potassium-pyridinedithiocarbazate (III) was produced by overnight agitation of compound (II) with CS2 in presence of ethanolic KH and later on compound (III) was refluxed with 99% NH 2 NH 2.H 2, neutralized by 50% HCl to acquire 4-amino -5-(4-aminophenyl)-4H-1,2,4-triazole-3-thiol (IV). Target compounds 4 - [ 6-aryl - 7H - [ 1,2,4 ] triazolo [ 3,4-b ] [1,3,4]-thiadiazine-3-yl] aniline (Va-k) were synthesized by condensation of compound (IV) with various α-bromo ketone in ethanol. The IR spectra using KBr pellet method were recorded, which divulge that functional groups appeared in the target compounds at their characteristic frequency; C-Cl, aromatic, str. between cm-1, C- -C, ring, str. at 1183 cm-1, C=N, aromatic, str. between cm-1, C-N2, aromatic, str. between cm-1, C=C, aromatic, str. between cm- 1,C=, chromen, str. at 1715cm-1, C-NH 2 str. between cm-1, C H, aromatic, str. between cm-1, H, str. between cm-1. The chemical shift (δ) for methyl hydrogen was observed at 2.27 ppm, δ value for methoxy hydrogen was observed at 3.92 ppm, methylene hydrogen was observed in the range of ppm, amino hydrogen in the range of ppm, aromatic hydrogen in the range of ppm, hydroxyl hydrogen in the range of ppm.the m/e value for compounds Va-k was observed between (M)+. The physical and spectral data indicate that the desired compounds were synthesized successfully. 3.2 Antimicrobial evaluation The in vitro antimicrobial evaluation of target compounds Va-k were carried out against two gram-positive bacteria species, namely Staphylococcus aureus and Bacillus subtilis, and two gram-negative bacteria species, namely Escherichia coli and Pseudomonas aeruginosa. Also compounds Va-k were evaluated for their in vitro antifungal evaluation against two fungi species, namely Aspergillus niger and Candida albicans. Minimum inhibitory concentration was determined by broth dilution method for all target compounds Va-k. Norfloxacin and the Ketoconazole were used as references standards. The results are represented in Tables 1. The results indicate that the in vivo antibacterial evaluation of newly synthesized target compound Ve was found to be most formidable with broad spectrum activity with MIC of 63, Published by DYPIPSR, Pimpri, Pune ( MH ) INDIA 33

5 77, 78 and 116, against Staphylococcus aureus, Escherichia coli, Bacillus subtilis, and Pseudomonas aeruginosa respectively. Compound substituted with o- H, on phenyl ring (Va) was found most significant agent among the newly synthesized series. Also target compound Ve showed most formidable and broad spectrum antifungal activity with MIC of 94 and 98 against Aspergillus niger and C.albicans respectively.compounds substituted with p- N2, p-nh2, m-n2 and aryl substitution on phenyl ring (Vg, Vh, Vi and Vk) were found significant agents in newly synthesised series. 3.3 Anticancer evaluation Cytotoxicity of synthesized compounds Va, Vb, Vc, Vf, and Vj were evaluated at concentration 10-5 M against set of 60 human cancer cell lines. All the tested five compounds were found to possess broad spectrum activity against selected cell lines (Tables 2). Compound Va, Vb, Vc, Vf and Vj were found to exhibit high growth inhibitory activity against Non-Small Cell Lung Cancer (HP-92, NCI-H23), Breast Cancer (MCF7, MDA-MB-468, MDA-MB-231/ATCC,BT-549), Renal cancer (U-31), CNS Cancer (SNB-19), Leukemia (CCRF-CEM), varian Cancer (VCAR-3) respectively. A growth % of most sensitive cell line to be , the mean growth % and fall in a range to for compound Va, A growth % of most sensitive cell line to be and the mean growth % and fall in a range to for compound Vb, A growth % of most sensitive cell line to be and the mean growth % and fall in a range to for compound Vc, A growth % of most sensitive cell line to be and the mean growth % and fall in a range to for compound Vf and A growth % of most sensitive cell line to be and the mean growth % and fall in a range to for compound Vj. 4. Conclusion Compounds having hydroxyl, chloro, methoxy, methyl functional group substituted on phenyl ring and chromen attached to [1,2,4]triazolo[3,4-b][1,3,4]-thiadiazine-3-yl nucleus are most significant antimicrobial and anticancer agent; these compounds could be used for further development to obtain more promising drug candidates. Table 1. Minimum inhibitory concentration values (µg/ml) for compounds Va-k against test microorganisms. Comp R. E. P. S. B. A. C. coli aeruginosa aureus subtilis niger Albicans Va o-h-ph Vb p-h-ph Vc p-cl-ph Vd p-br-ph Ve P-CH 3 -ph Vf p-ch 3 -ph Vg p-n 2 -ph Vh p-nh 2 -ph Vi m-n 2 -ph Vj 2H-chromen-2-one Vk Bi-ph Norfloxacin Ketoconazole Published by DYPIPSR, Pimpri, Pune ( MH ) INDIA 34

6 Table 2. Anticancer screening data of five tested compounds Concn1.00E-5 molar Comp Mean Growth % Range Growth % Most Sensative Cell Lines Growth of Most Sensitive Cell Lines Va Non-Small Cell Lung Cancer (HP-92) Va Non-Small Cell Lung Cancer (NCI-H23) Va Breast Cancer (MDA-MB-468) Vb Renal Cancer (U-31) Vb CNS Cancer (SNB-19) Vb Breast Cancer (MDA-MB-231/ATCC) Vc Renal Cancer (U-31) Vc Breast Cancer (MCF7) Vc Breast Cancer (MDA-MB-468) Vf Leukemia (CCRF-CEM) Vf Leukemia (SR) Vf Breast Cancer (BT-549) IVj varian Cancer (VCAR-3) IVj Renal Cancer (U-31) IVj Breast Cancer (BT-549) Acknowledgement The authors thank to Dr. V. K. Deshmukh, Principal, MES`s College of Pharmacy, Sonai for providing all laboratory facilities, NCI, Maryland, USA for carrying out cell line inhibitory activity, Macleod s Pharma. Ltd., Daman, India for providing gift samples, UDCT Dr. BAMU, Aurangabad for recording FT-IR Spectra, 1 H- NMR spectra at SAIF Punjab University, Chandigarh, GC -MS spectra at Savitribai Phule Pune University, Pune for recording spectra. References [1] Holla S. B., Poojary K. N., Rao B. S., Shivananda M. K. New bis-aminomer -captotriazoles and bis - triazolothiadiazoles as possible anticancer agents. Eur. J. Med. Chem. 2002, 37, [2] Bhat, K., Poojary, B., Prasad, D., Naik, P. and Holla, B. Synthesis and antitumor activity studies of some new fused 1,2,4-triazole derivatives carrying 2,4-dichloro-5-fluorophenyl moiety. European Journal of Medicinal Chemistry, 2009, 44(12), [3] Hu, M., Zhang, B., Yang, H., Liu, Y., Chen, Y., Ma, T., Lu, L., You, W. and Zhao, P. Design, Synthesis and Molecular Docking Studies of Novel Indole- Pyrimidine Hybrids as Tubulin Polymerization Inhibitors. Chemical Biology & Drug Design, 2015, 86(6), [4] Karegoudarb P., Jagdeesh Prasada D., Ashoka M., Mahalingab M., Poojarya B., Holla B. S. Synthesis, antimicrobial and anti-inflammatory activities of some 1,2,4-triazolo[3,4-b][1,3,4]thiadiazoles and 1,2,4-triazolo[3,4-b][1,3,4]thiadiazines bearing trichlorophenyl moiety. Eur. J Med. Chem. 2008, 43 (4), [5] Karabasanagouda T., Adhikari A. V., Shetty N. S. Synthesis and antimicrobial activities of some novel 1,2,4-triazolo[3,4-b]-1,3,4-thiadiazoles and 1,2,4- Published by DYPIPSR, Pimpri, Pune ( MH ) INDIA 35

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